Rnai agents for inhibiting expression of androgen receptor (AR), compositions thereof, and methods of use

Abstract

Described are RNAi agents, compositions that include RNAi agents, and methods for inhibition of an Androgen Receptor (AR) gene. The AR RNAi agents and RNAi agent conjugates disclosed herein inhibit the expression of an AR gene. Pharmaceutical compositions that include one or more AR RNAi agents, optionally with one or more additional therapeutics, are also described. Delivery of the described AR RNAi agents to central nervous system (CNS) tissue and / or skeletal muscle tissue, in vivo, provides for inhibition of AR gene expression and a reduction in AR activity, which can provide a therapeutic benefit to subjects, including human subjects, for the treatment of various diseases including spinal and bulbar muscular atrophy (SBMA).

Description

RNAi Agents for Inhibiting Expression of Androgen Receptor (AR), Compositions Thereof, And Methods of UseCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority from United States Provisional Patent Application Serial No. 63 / 624,417, filed on January 24, 2024, United States Provisional Patent Application Serial No. 63 / 573, 151, filed April 2, 2024. United States Provisional Patent Application Serial No. 63 / 662,243, filed June 20, 2024, United States Provisional Patent Application Serial No. 63 / 718,062, filed November 8, 2024, and United States Provisional Patent Application Serial No. 63 / 724,015, filed November 22, 2024, the contents of each of which are incorporated herein by reference in their entirety .SEQUENCE LISTING

[0002] This application contains a Sequence Listing (in compliance with Standard ST26), which has been submitted in xml format and is hereby incorporated by reference in its entirety. The xml sequence listing file is named 30710-WO SeqListing.xml, created January 23, 2025, and is 3922 kb in size.FIELD OF THE INVENTION

[0003] The present disclosure relates to RNA interference (RNAi) agents, e.g., double stranded RNAi agents, for inhibition of Androgen Receptor (“AR”) gene expression, compositions that include AR RNAi agents, and methods of use thereof.BACKGROUND

[0004] The androgen receptor (“AR”) is a type of nuclear receptor activated by binding of androgenic hormones, such as testosterone. In some males, trinucleotide CAG repeat expansions in exon 1 of the androgen receptor gene results in loss of AR function and a toxic gain of function related to lower motor neuron disease, called spinal and bulbar muscular atrophy (“SBMA”), also known as Kennedy’s disease.

[0005] SBMA is an adult-onset disease that slowly progresses over several years, causing bulbar and lower motor neuron loss, and muscle denervation. The loss of bulbar neurons may cause dysarthria and dysphagia. There is currently no cure for SBMA, and current treatmentfocuses on preventing disease complications. Therefore there exists a need for treatments that reduce mutant AR gene production.SUMMARY

[0006] There exists a need for novel RNA interference (RNAi) agents (termed RNAi agents, RNAi triggers, or triggers), e.g., double stranded RNAi agents, that are able to selectively and efficiently inhibit the expression of an AR gene, including for use as a therapeutic or medicament. Further, there exists a need for compositions of novel AR-specific RNAi agents for the treatment of diseases or disorders associated mutant AR expression and / or disorders that can be mediated at least in part by a reduction in AR gene expression and / or AR receptor expression.

[0007] The nucleotide sequences and chemical modifications of the AR RNAi agents disclosed herein, as well as their combination with certain specific targeting ligands suitable for selectively and efficiently delivering the AR RNAi agents to relevant CNS and / or skeletal muscle cells in vivo, differ from those previously disclosed or known in the art. The AR RNAi agents disclosed herein provide for highly potent and efficient inhibition of the expression of an AR gene.

[0008] The nucleotide sequences and chemical modifications of the AR RNAi agents disclosed herein, as well as their combination with certain specific antigen binding proteins suitable for selectively and efficiently delivering the AR RNAi agents to relevant CNS cells in vivo, differ from those previously disclosed or known in the art. The AR RNAi agents disclosed herein provide for highly potent and efficient inhibition of the expression of an AR gene.

[0009] In general, the present disclosure features AR gene-specific RNAi agents, compositions that include AR RNAi agents, and methods for inhibiting expression of an AR gene in vitro and / or in vivo using the AR RNAi agents and compositions that include AR RNAi agents described herein. The AR RNAi agents described herein are able to selectively and efficiently decrease expression of an AR gene, and thereby reduce the expression of the androgen receptor.

[0010] The described AR RNAi agents can be used in methods for therapeutic treatment (including preventative or prophylactic treatment) of symptoms and diseases including, but not limited to various neuromuscular diseases (including SBMA or Kennedy’s disease.)

[0011] In one aspect, the disclosure features RN Ai agents for inhibiting expression of an AR (or AR) gene, wherein the RNAi agent includes a sense strand (also referred to as a passenger strand) and an antisense strand (also referred to as a guide strand). The sense strand and theantisense strand can be partially, substantially, or fully complementary to each other. The length of the RNAi agent sense and antisense strands described herein each can be 16 to 49 nucleotides in length. In some embodiments, the sense and antisense strands are independently 17 to 26 nucleotides in length. The sense and antisense strands can be either the same length or different lengths. In some embodiments, the sense and antisense strands are independently 21 to 26 nucleotides in length. In some embodiments, the sense and antisense strands are independently 21 to 24 nucleotides in length. In some embodiments, both the sense strand and the antisense strand are 21 nucleotides in length. In some embodiments, the antisense strands are independently 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. In some embodiments, the sense strands are independently 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, or 49 nucleotides in length. The RNAi agents described herein, upon delivery to a cell expressing AR such as endothelial cells, neurons, microglia, and astrocytes, inhibit the expression of one or more AR gene variants in vivo and / or in vitro.

[0012] The AR RNAi agents disclosed herein target a human AR gene (see, e.g. SEQ ID NO: 1 ). In some embodiments, the AR RNAi agents disclosed herein target a portion of an AR gene ha ving the sequence of any of the sequences disclosed in Table 1.

[0013] In another aspect, the disclosure features compositions, including pharmaceutical compositions, that include one or more of the disclosed AR RNAi agents that are able to selectively and efficiently decrease expression of an AR gene. The compositions that include one or more AR RNAi agents described herein can be administered to a subject, such as a human or animal subject, for the treatment (including prophylactic treatment or inhibition) of symptoms and diseases associated with AR receptor activity.

[0014] Examples of AR RNAi agent sense strands and antisense strands that can be used in an AR RNAi agent are provided in Tables 3, 4, 5, and 6. Examples of AR RNAi agent duplexes are provided in Tables 7, 8, 9, and 10. Examples of 19-nucleotide core stretch sequences that may consist of or may be included in the sense strands and antisense strands of certain AR RNAi agents disclosed herein, are provided in Table 2.

[0015] In another aspect, the disclosure features methods for delivering AR RNAi agents to neurons, astrocytes, microglia and endothelial cells in a subject, such as a mammal, in vivo. Also described herein are compositions for use in such methods. In some embodiments, disclosed herein are methods for delivering AR RNAi agents to central nervous system cells (neurons, astrocytes, microglia and endothelial cells) to a subject in vivo. In some embodiments, the subject is a human subject

[0016] The methods disclosed herein include the administration of one or more AR RNAi agents to a subject, e.g., a human or animal subject, by any suitable means known in the art. The pharmaceutical compositions disclosed herein that include one or more AR RNAi agents can be administered m a number of ways depending upon whether local or systemic treatment is desired. Administration can be, but is not limited to, for example, intravenous, intraarterial, subcutaneous, intraperitoneal, subdermal (e.g., via an implanted device), and intraparenchymal administration. In some embodiments, the pharmaceutical compositions described herein are administered by intrathecal injection or intracerebroventricular injection.

[0017] In some embodiments, it is desired that the AR RNAi agents described herein inhibit the expression of an AR gene in central nervous system cells.

[0018] The one or more AR RNAi agents can be delivered to target cells or tissues using any oligonucleotide delivery technology known in the art. In some embodiments, an AR RNAi agent is delivered to cells or tissues by covalently linking the RNAi agent to a targeting group, lipid moiety, or antigen binding protein.

[0019] A targeting group can be linked to the 3' or 5' end of a sense strand or an antisense strand of an AR RNAi agent. In some embodiments, a targeting group is linked to the 3' or 5' end of the sense strand. In some embodiments, a targeting group is linked to the 5' end of the sense strand. In some embodiments, a targeting group is linked internally to a nucleotide on the sense strand and / or the antisense strand of the RNAi agent. In some embodiments, a targeting group is linked to the RNAi agent via a linker.

[0020] In another aspect, the disclosure features compositions that include one or more AR RNAi agents that have the duplex structures disclosed in Tables 7, 8, 9. and 10.

[0021] In some embodiments, an AR RNAi agent is linked to one or more linking groups or other non-nucleotide groups or compounds, such as pharmacokinetic / pharmacodynamic (PK / PD) modulators. PK / PD modulators can increase circulation time of the conjugated drug and / or increase the activity of the RNAi agent through improved cell receptor binding, improved cellular uptake, and / or other means. Examples of PK / PD modulators suitable for use with the AR RNAi agents disclosed herein can be found in Table 11, herein.

[0022] In some embodiments, an AR RNAi agent is conjugated to a targeting group, a linking group, a PK / PD modulator, and / or another non-nucleotide group. In some embodiments, an AR RNAi agent is conjugated to a targeting group and a PK / PD modulator.

[0023] The use of AR RNAi agents provides methods for therapeutic (including prophylactic) treatment of diseases or disorders for which a reduction in AR receptor activity can provide atherapeutic benefit. The AR RNAi agents disclosed herein can be used to treat various neurodegenerative diseases, including SBMA. Such methods of treatment include administration of an AR RNAi agent to a human being or animal having mutant androgen receptor or androgen receptor activity beyond desirable levels.

[0024] The one or more AR RNAi agents can be delivered to target cells or tissues using any oligonucleotide delivery technology known in the art. In some embodiments, an AR RNAi agent is delivered to cells or tissues by covalently linking the RNAi agent to a targeting group or an antigen binding protein.

[0025] The one or more AR RNAi agents can be delivered to target cells or tissues using any oligonucleotide delivery technology known in the art. In some embodiments, an AR RNAi agent is delivered to cells or tissues by covalently linking the RNAi agent to a targeting group. In some embodiments, the targeting group can include a cell receptor ligand. A targeting group can be linked to the 3' or 5' end of a sense strand or an antisense strand of an AR RNAi agent, or can be linked via one or more internal nucleotides of the sense strand or the antisense strand. In some embodiments, a targeting group is linked to the 3' or 5' end of the sense strand. In some embodiments, a targeting group is linked to the 5' end of the sense strand. In some embodiments, a targeting group is linked internally to a nucleotide on the sense strand and / or the antisense strand of the RNAi agent. In some embodiments, a targeting group is linked to the RNAi agent via a linker. Example targeting ligands suitable for use that have affinity for skeletal muscle cells and / or receptors present on skeletal muscle cells (e.g., integrin alpha-v- beta-6 (av|36)), are shown in Table 1 1 herein.

[0026] An antigen binding protein can be linked to the 3' or 5' end of a sense strand or an antisense strand of an AR RNAi agent. In some embodiments, an antigen binding protein is linked to the 3' or 5' end of the sense strand. In some embodiments, an antigen binding protein is linked to the 5' end of the sense strand. In some embodiments, an antigen binding protein is linked internally to a nucleotide on the sense strand and / or the antisense strand of the RNAi agent. In some embodiments, an antigen binding protein is linked to the RNAi agent via a linker.Definitions

[0027] As used herein, the terms ‘’oligonucleotide” and ‘’polynucleotide” mean a polymer of linked nucleosides each of which can be independently modified or unmodified.

[0028] As used herein, an “RNAi agent" (also referred to as an “RNAi trigger”) means a composition that contains an RNA or RNA-like (e.g., chemically modified RNA) oligonucleotide molecule that is capable of degrading or inhibiting (e.g., degrades or inhibits under appropriate conditions) translation of messenger RNA (mRNA) transcripts of a target mRNA in a sequence specific manner. As used herein, RNAi agents may operate through the RNA interference mechanism (i.e., inducing RNA interference through interaction with the RNA interference pathway machinery (RNA-induced silencing complex or RISC) of mammalian cells), or by any alternative mechanism(s) or pathway(s). While it is believed that RNAi agents, as that term is used herein, operate primarily through the RNA interference mechanism, the disclosed RNAi agents are not bound by or limited to any particular pathway or mechanism of action. RNAi agents disclosed herein are comprised of a sense strand and an antisense strand, and include, but are not limited to: short (or small) interfering RNAs (siRNAs), double stranded RNAs (dsRNA), micro RNAs (miRNAs), short hairpin RNAs (shRNA), and dicer substrates. The antisense strand of the RNAi agents described herein is at least partially complementary to the mRNA being targeted (i.e. AR mRNA). RNAi agents can include one or more modified nucleotides and / or one or more non-phosphodiester linkages.

[0029] As used herein, the terms “silence,” “reduce,” “inhibit,” “down-regulate,” or “knockdown” when referring to expression of a given gene, mean that the expression of the gene, as measured by the level of RNA transcribed from the gene or the level of polypeptide, protein, or protein subunit translated from the mRNA in a cell, group of cells, tissue, organ, or subject in which the gene is transcribed, is reduced when the cell, group of cells, tissue, organ, or subject is treated with the RNAi agents described herein as compared to a second cell, group of cells, tissue, organ, or subject that has not or have not been so treated.

[0030] As used herein, the terms “sequence” and “nucleotide sequence” mean a succession or order of nucleobases or nucleotides, described with a succession of letters using standard nomenclature.

[0031] As used herein, a “base,” “nucleotide base,” or “nucleobase,” is a heterocyclic pyrimidine or purine compound that is a component of a nucleotide, and includes the primary purine bases adenine and guanine, and the primary' pyrimidine bases cytosine, thymine, and uracil. A nucleobase may further be modified to include, without limitation, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases. (See. e.g., Modified Nucleosides in Biochemistry, Biotechnology' and Medicine, Herdewijn, P. ed. Wiley - VCH, 2008). The synthesis of such modified nucleobases (including phosphoramidite compounds that include modified nucleobases) is known in the art.

[0032] As used herein, and unless otherwise indicated, the term “complementary,’' when used to describe a first nucleobase or nucleotide sequence (e.g.. RNAi agent sense strand or targeted mRNA) in relation to a second nucleobase or nucleotide sequence (e.g., RN Ai agent antisense strand or a single-stranded antisense oligonucleotide), means the ability of an oligonucleotide or polynucleotide including the first nucleotide sequence to hybridize (form base pair hydrogen bonds under mammalian physiological conditions (or otherwise suitable in vivo or in vitro conditions) and form a duplex or double helical structure under certain standard conditions with an oligonucleotide that includes the second nucleotide sequence. The person of ordinary skill in the art would be able to select the set of conditions most appropriate for a hybridization test. Complementary sequences include Watson-Crick base pairs or non-Watson-Crick base pairs and include natural or modified nucleotides or nucleotide mimics, at least to the extent that the above hybridization requirements are fulfilled. Sequence identity or complementarity is independent of modification. For example, a and Af, as defined herein, are complementary to U (or T) and identical to A for the purposes of determining identity or complementarity.

[0033] As used herein, “perfectly complementary” or “fully complementary” means that in a hybridized pair of nucleobase or nucleotide sequence molecules, all (100%) of the bases in a contiguous sequence of a first oligonucleotide will hybridize with the same number of bases in a contiguous sequence of a second oligonucleotide. The contiguous sequence may comprise all or a part of a first or second nucleotide sequence.

[0034] As used herein, “partially complementary ” means that in a hybridized pair of nucleobase or nucleotide sequence molecules, at least 70%, but not all, of the bases in a contiguous sequence of a first oligonucleotide will hy bridize with the same number of bases in a contiguous sequence of a second oligonucleotide. The contiguous sequence may comprise all or a part of a first or second nucleotide sequence.

[0035] As used herein, “substantially complementary” means that in a hybridized pair of nucleobase or nucleotide sequence molecules, at least 85%, but not all, of the bases in a contiguous sequence of a first oligonucleotide will hybridize with the same number of bases in a contiguous sequence of a second oligonucleotide. The contiguous sequence may comprise all or a part of a first or second nucleotide sequence.

[0036] As used herein, the terms “complementary,” “fully complementary ,” “partially complementary',” and “substantially complementary ” are used with respect to the nucleobase or nucleotide matching between the sense strand and the antisense strand of an RNAi agent, or between the antisense strand of an RNAi agent and a sequence of an AR mRNA.

[0037] As used herein, the term “substantially identical’" or “substantial identity,” as applied to a nucleic acid sequence means the nucleotide sequence (or a portion of a nucleotide sequence) has at least about 85% sequence identity or more, e.g., at least 90%, at least 95%, or at least 99% identity, compared to a reference sequence. Percentage of sequence identity is determined by comparing two optimally aligned sequences over a comparison window. The percentage is calculated by determining the number of positions at which the same type of nucleic acid base occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions m the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity. The inventions disclosed herein encompass nucleotide sequences substantially identical to those disclosed herein.

[0038] As used herein, the terms “treat,"’ “treatment,” and the like, mean the methods or steps taken to provide relief from or alleviation of the number, severity', and / or frequency of one or more symptoms of a disease in a subject. As used herein, “treat” and “treatment” may include the prevention, management, prophylactic treatment, and / or inhibition or reduction of the number, severity, and / or frequency of one or more symptoms of a disease in a subject.

[0039] As used herein, the phrase “introducing into a cell,” when referring to an RNAi agent, means functionally delivering the RNAi agent into a cell. The phrase “functional delivery,” means delivering the RNAi agent to the cell in a manner that enables the RNAi agent to have the expected biological activity, e.g., sequence-specific inhibition of gene expression.

[0040] Unless stated otherwise, use of the symbolas used herein means that any group or groups may be linked thereto that is in accordance with the scope of the inventions described herein.

[0041] As used herein, the term “isomers” refers to compounds that have identical molecular formulae, but that differ in the nature or the sequence of bonding of their atoms or in the arrangement of their atoms in space. Isomers that differ in the arrangement of their atoms in space are temied “stereoisomers.” Stereoisomers that are not mirror images of one another are termed “diastereoisomers,"’ and stereoisomers that are non-superimposable mirror images are termed “enantiomers,” or sometimes optical isomers. A carbon atom bonded to four nonidentical substituents is termed a “chiral center.”

[0042] As used herein, unless specifically identified in a structure as having a particular conformation, for each structure in which asymmetric centers are present and thus give rise to enantiomers, diastereomers, or other stereoisomeric configurations, each structure disclosedherein is intended to represent all such possible isomers, including their optically pure and racemic forms. For example, the structures disclosed herein are intended to cover mixtures of diastereomers as well as single stereoisomers.

[0043] As used in a claim herein, the phrase “consisting of’ excludes any element, step, or ingredient not specified in the claim. When used in a claim herein, the phrase “consisting essentially of’ limits the scope of a claim to the specified materials or steps and those that do not materially affect the basic and novel characteristic(s) of the claimed invention.

[0044] The person of ordinary' skill in the art would readily understand and appreciate that the compounds and compositions disclosed herein may have certain atoms (e g., N, O, or S atoms) in a protonated or deprotonated state, depending upon the environment in which the compound or composition is placed. Accordingly, as used herein, the structures disclosed herein envisage that certain functional groups, such as, for example, OH, SH, or NH, may be protonated or deprotonated. The disclosure herein is intended to cover the disclosed compounds and compositions regardless of their state of protonation based on the environment (such as pH), as would be readily understood by the person of ordinary skill in the art. Correspondingly, compounds described herein with labile protons or basic atoms should also be understood to represent salt forms of the corresponding compound. Compounds described herein may be in a free acid, free base, or salt form. Pharmaceutically acceptable salts of the compounds described herein should be understood to be within the scope of the invention.

[0045] As used herein, the term “linked” or “conjugated” when referring to the connection between two compounds or molecules means that two compounds or molecules are joined by a covalent bond. Unless stated, the terms “linked” and “conjugated” as used herein may refer to the connection between a first compound and a second compound either with or without any intervening atoms or groups of atoms.

[0046] As used herein, the term “including” is used to herein mean, and is used interchangeably with, the phrase “including but not limited to.” The term “or” is used herein to mean, and is used interchangeably with, the term “and / or,” unless the context clearly indicates otherwise.

[0047] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary' skill in the art. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below'. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions,will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0048] Other objects, features, aspects, and advantages of the invention will be apparent from the following detailed description, accompanying figures, and from the claims.DETAILED DESCRIPTIONRNAi Agents

[0049] Described herein are RNAi agents for inhibiting expression of the AR gene (referred to herein as AR RNAi agents or AR RNAi triggers). Each AR RNAi agent disclosed herein comprises a sense strand and an antisense strand. The sense strand and the antisense strand each can be 16 to 49 nucleotides in length. The sense and antisense strands can be either the same length or they can be different lengths. In some embodiments, the sense and antisense strands are each independently 18 to 27 nucleotides in length. In some embodiments, both the sense and antisense strands are each 21-26 nucleotides in length. In some embodiments, the sense and antisense strands are each 21-24 nucleotides in length. In some embodiments, the sense and antisense strands are each independently 19-21 nucleotides in length. In some embodiments, the sense strand is about 19 nucleotides in length while the antisense strand is about 21 nucleotides in length. In some embodiments, the sense strand is about 21 nucleotides in length while the antisense strand is about 23 nucleotides in length. In some embodiments, a sense strand is 23 nucleotides in length and an antisense strand is 21 nucleotides in length. In some embodiments, both the sense and antisense strands are each 21 nucleotides in length. In some embodiments, the RNAi agent sense and antisense strands are each independently 16. 17. 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32. 33, 34, 35, 36, 37, 38, or 39 nucleotides in length. In some embodiments, a double-stranded RNAi agent has a duplex length of about 16, 17, 18, 19, 20, 21, 22, 23 or 24 nucleotides.

[0050] Examples of nucleotide sequences used in forming AR RNAi agents are provided in Tables 2, 3, 4, 5, 6. and 10. Examples of RNAi agent duplexes, that include the sense strand and antisense strand sequences in Tables 2, 3, 4, 5, 6, are shown in Tables 7, 8, 9, and 10.

[0051] In some embodiments, the region of perfect, substantial, or partial complementarity between the sense strand and the antisense strand is 16-26 (e.g., 16, 17. 18. 19. 20. 21, 22, 23, 24, 25, or 26) nucleotides in length and occurs at or near the 5' end of the antisense strand (e.g., this region may be separated from the 5' end ofthe antisense strand by 0, 1, 2. 3, or 4 nucleotides that are not perfectly, substantially, or partially complementary).

[0052] A sense strand of the AR RNAi agents described herein includes at least 16 consecutive nucleotides that have at least 85% identity to a core stretch sequence (also referred to herein as a “core stretch” or “core sequence”) of the same number of nucleotides in an AR mRNA. In some embodiments, a sense strand core stretch sequence is 100% (perfectly) complementary or at least about 85% (substantially) complementary to a core stretch sequence in the antisense strand, and thus the sense strand core stretch sequence is typically perfectly identical or at least about 85% identical to a nucleotide sequence of the same length (sometimes referred to, e.g. as a target sequence) present in the AR mRNA target. In some embodiments, this sense strand core stretch is 16, 17, 18, 19, 20, 21, 22, or 23 nucleotides in length. In some embodiments, this sense strand core stretch is 17 nucleotides in length. In some embodiments, this sense strand core stretch is 19 nucleotides in length.

[0053] An antisense strand of an AR RNAi agent described herein includes at least 16 consecutive nucleotides that have at least 85% complementarity to a core stretch of the same number of nucleotides in an AR mRNA and to a core stretch of the same number of nucleotides in the corresponding sense strand. In some embodiments, an antisense strand core stretch is 100% (perfectly) complementary or at least about 85% (substantially) complementary to a nucleotide sequence (e.g, target sequence) of the same length present in the AR mRNA target. In some embodiments, this antisense strand core stretch is 16, 17, 18, 19, 20, 21, 22, or 23 nucleotides in length. In some embodiments, this antisense strand core stretch is 19 nucleotides in length. In some embodiments, this antisense strand core stretch is 17 nucleotides in length. A sense strand core stretch sequence can be the same length as a corresponding antisense core sequence or it can be a different length.

[0054] The AR RNAi agent sense and antisense strands anneal to form a duplex. A sense strand and an antisense strand of an AR RNAi agent can be partially, substantially, or fully complementary to each other. Within the complementary duplex region, the sense strand core stretch sequence is at least 85% complementary or 100% complementary to the antisense core stretch sequence. In some embodiments, the sense strand core stretch sequence contains a sequence of at least 16, at least 17. at least 18, at least 19, at least 20, at least 21, at least 22, or at least 23 nucleotides that is at least 85% or 100% complementary to a corresponding 16. 17. 18, 19, 20, 21 , 22, or 23 nucleotide sequence of the antisense strand core stretch sequence (i.e., the sense and antisense core stretch sequences of an AR RNAi agent have a region of at least 16, at least 17, at least 18, at least 19, at least 20, at least 21 , at least 22, or at least 23 nucleotides that is at least 85% base paired or 100% base paired.)

[0055] In some embodiments, the antisense strand of an AR RNAi agent disclosed herein differs by 0, 1. 2, or 3 nucleotides from any of the antisense strand sequences in Table 2 or Table 3. In some embodiments, the sense strand of an AR RNAi agent disclosed herein differs by 0, 1, 2, or 3 nucleotides from any of the sense strand sequences in Table 2, Table 4, Table 5, Table 6, or Table 10.

[0056] In some embodiments, the sense strand and / or the antisense strand can optionally and independently contain an additional 1. 2, 3, 4. 5, or 6 nucleotides (extension) at the 3' end. the 5' end, or both the 3' and 5' ends of the core stretch sequences. The antisense strand additional nucleotides, if present, may or may not be complementary' to the corresponding sequence in the AR mRNA The sense strand additional nucleotides, if present, may or may not be identical to the corresponding sequence in the AR mRNA. The antisense strand additional nucleotides, if present, may or may not be complementary to the corresponding sense strand’s additional nucleotides, if present.

[0057] As used herein, an extension comprises 1, 2, 3, 4, 5, or 6 nucleotides at the 5' and / or 3' end of the sense strand core stretch sequence and / or antisense strand core stretch sequence. The extension nucleotides on a sense strand may or may not be complementary to nucleotides, either core stretch sequence nucleotides or extension nucleotides, in the corresponding antisense strand. Conversely, the extension nucleotides on an antisense strand may or may not be complementary to nucleotides, either core stretch nucleotides or extension nucleotides, in the corresponding sense strand. In some embodiments, both the sense strand and the antisense strand of an RNAi agent contain 3' and 5' extensions. In some embodiments, one or more of the 3' extension nucleotides of one strand base pairs with one or more 5' extension nucleotides of the other strand. In other embodiments, one or more of 3' extension nucleotides of one strand do not base pair with one or more 5' extension nucleotides of the other strand. In some embodiments, an AR RNAi agent has an antisense strand having a 3' extension and a sense strand having a 5' extension. In some embodiments, the extension nucleotide(s) are unpaired and form an overhang. As used herein, an “overhang” refers to a stretch of one or more unpaired nucleotides located at a terminal end of either the sense strand or the antisense strand that does not form part of the hybridized or duplexed portion of an RNAi agent disclosed herein.

[0058] In some embodiments, an AR RNAi agent comprises an antisense strand having a 3' extension of 1, 2, 3, 4, 5, or 6 nucleotides in length. In other embodiments, an AR RNAi agent comprises an antisense strand having a 3' extension of I, 2, or 3 nucleotides in length. In some embodiments, one or more of the antisense strand extension nucleotides comprise nucleotides that are complementary to the corresponding AR mRNA sequence. In some embodiments, oneor more of the antisense strand extension nucleotides comprise nucleotides that are not complementary to the corresponding AR mRNA sequence.

[0059] In some embodiments, an AR RNAi agent comprises a sense strand having a 3' extension of 1, 2, 3, 4, or 5 nucleotides in length. In some embodiments, one or more of the sense strand extension nucleotides comprises adenosine, uracil, or thymidine nucleotides, AT dinucleotide, or nucleotides that correspond to or are the identical to nucleotides in the AR mRNA sequence. In some embodiments, the 3' sense strand extension includes or consists of one of the following sequences, but is not limited to: T, UT, TT, UU, UUT, TTT, or TTTT (each listed 5' to 3').

[0060] A sense strand can have a 3' extension and / or a 5' extension. In some embodiments, an AR RNAi agent comprises a sense strand having a 5' extension of 1, 2, 3, 4. 5, or 6 nucleotides in length. In some embodiments, one or more of the sense strand extension nucleotides comprise nucleotides that correspond to or are identical to nucleotides in the AR mRNA sequence.

[0061] Examples of sequences used in forming AR RNAi agents are provided in Tables 2, 3, 4, 5, 6. and 10. In some embodiments, an AR RNAi agent antisense strand includes a sequence of any of the sequences in Tables 2, 3, or 10. In certain embodiments, an AR RNAi agent antisense strand comprises or consists of any one of the modified sequences in Table 3. In some embodiments, an AR RNAi agent antisense strand includes the sequence of nucleotides (from 5' end -» 3' end) 1-17. 2-15. 2-17. 1 -18. 2-18. 1-19. 2-19, 1-20, 2-20, 1-21, or 2-21, of any of the sequences in Tables 2 or 3. In some embodiments, an AR RNAi agent sense strand includes the sequence of any of the sequences in Tables 2, 4. 5, or 6. In some embodiments, an AR RNAi agent sense strand includes the sequence of nucleotides (from 5' end -> 3' end) 1-18, 1- 19, 1-20, 1-21. 2-19. 2-20. 2-21. .3-20. 3-21. or 4-21 of any of the sequences in Tables 2, 4, 5, or 6. In certain embodiments, an AR RNAi agent sense strand comprises or consists of a modified sequence of any one of the modified sequences in Table 4, 5, 6, or 10.

[0062] In some embodiments, the sense and antisense strands of the RNAi agents described herein contain the same number of nucleotides. In some embodiments, the sense and antisense strands of the RNAi agents described herein contain different numbers of nucleotides. In some embodiments, the sense strand 5' end and the antisense strand 3' end of an RNAi agent form a blunt end. In some embodiments, the sense strand 3' end and the antisense strand 5' end of an RNAi agent form a blunt end. In some embodiments, both ends of an RNAi agent form blunt ends. In some embodiments, neither end of an RNAi agent is blunt-ended. As used herein a“blunt end” refers to an end of a double stranded RNAi agent in which the terminal nucleotides of the two annealed strands are complementary (form a complementary base-pair).

[0063] In some embodiments, the sense strand 5' end and the antisense strand 3' end of an RNAi agent form a frayed end. In some embodiments, the sense strand 3' end and the antisense strand 5 ' end of an RNAi agent form a fray ed end. In some embodiments, both ends of an RNAi agent form a frayed end. In some embodiments, neither end of an RNAi agent is a frayed end. As used herein a frayed end refers to an end of a double stranded RNAi agent in which the terminal nucleotides of the two annealed strands form a pair (i.e., do not form an overhang) but are not complementary (i.e. form a non-compl ementary pair). In some embodiments, one or more unpaired nucleotides at the end of one strand of a double stranded RNAi agent form an overhang. The unpaired nucleotides may be on the sense strand or the antisense strand, creating either 3’ or 5' overhangs. In some embodiments, the RNAi agent contains: a blunt end and a frayed end, a blunt end and 5' overhang end, a blunt end and a 3' overhang end, a frayed end and a 5' overhang end, a frayed end and a 3' overhang end, two 5' overhang ends, two 3' overhang ends, a 5' overhang end and a 3' overhang end, two frayed ends, or two blunt ends. Typically, when present, overhangs are located at the 3’ terminal ends of the sense strand, the antisense strand, or both the sense strand and the antisense strand.

[0064] The AR RNAi agents disclosed herein may also be comprised of one or more modified nucleotides. In some embodiments, substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand of the AR RNAi agent are modified nucleotides. The AR RNAi agents disclosed herein may further be comprised of one or more modified intemucleoside linkages, e.g., one or more phosphorothioate linkages. In some embodiments, an AR RNAi agent contains one or more modified nucleotides and one or more modified intemucleoside linkages. In some embodiments, a 2'-modified nucleotide is combined with modified intemucleoside linkage.

[0065] In some embodiments, an AR RNAi agent is prepared or provided as a salt, mixed salt, or a free-acid. In some embodiments, an AR RNAi agent is prepared as a pharmaceutically acceptable salt. In some embodiments, an AR RNAi agent is prepared as a pharmaceutically acceptable sodium salt. Such forms that are well known in the art are within the scope of the inventions disclosed herein.Modified Nucleotides

[0066] Modified nucleotides, when used in various oligonucleotide constructs, can preserve activity of the compound in cells while at the same time increasing the serum stability of thesecompounds, and can also minimize the possibility of activating interferon activity in humans upon admini stration of the oligonucleotide construct.

[0067] In some embodiments, an AR RNAi agent contains one or more modified nucleotides. As used herein, a “modified nucleotide” is a nucleotide other than a ribonucleotide (2'-hydroxyl nucleotide). In some embodiments, at least 50% (e.g., at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100%) of the nucleotides are modified nucleotides. As used herein, modified nucleotides can include, but are not limited to, deoxyribonucleotides, nucleotide mimics, abasic nucleotides, 2'-modified nucleotides, inverted nucleotides, modified nucleobase-comprising nucleotides, bridged nucleotides, peptide nucleic acids (PNAs), 2',3'-seco nucleotide mimics (unlocked nucleobase analogues), locked nucleotides, 3'-O-methoxy (2' intemucleoside linked) nucleotides, 2'-F-Arabino nucleotides, 5'-Me, 2'-fluoro nucleotide, morpholino nucleotides, vinyl phosphonate deoxyribonucleotides, vinyl phosphonate containing nucleotides, and cyclopropyl phosphonate containing nucleotides. 2'-modified nucleotides (i.e., a nucleotide with a group other than a hydroxyl group at the 2' position of the five-membered sugar ring) include, but are not limited to. 2'-O-inethyl nucleotides (also referred to as 2'-methoxy nucleotides), 2'-fluoro nucleotides (also referred to herein as 2'-deoxy-2'-fluoro nucleotides), 2'-deoxy nucleotides, 2'- methoxyethyl (2'-O-2-methoxylethyl) nucleotides (also referred to as 2'-MOE), 2'-amino nucleotides, and 2'-alkyl nucleotides. It is not necessary for all positions in a given compound to be uniformly modified. Conversely, more than one modification can be incorporated in a single AR RNAi agent or even in a single nucleotide thereof. The AR RNAi agent sense strands and antisense strands can be synthesized and / or modified by methods known in the art. Modification at one nucleotide is independent of modification at another nucleotide.

[0068] Modified nucleobases include synthetic and natural nucleobases, such as 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted punnes, (e.g., 2-aminopropyladenine, 5-propynyluracil, or 5-propynylcytosine), 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, inosine, xanthine, hypoxanthine, 2-aminoadenine, 6-alkyl (e.g., 6- methyl, 6-ethyl, 6-isopropyl. or 6-n-butyl) derivatives of adenine and guanine, 2-alkyl (e.g., 2- methyl, 2-ethyl, 2 -isopropyl, or 2-n-butyl) and other alkyl derivatives of adenine and guanine. 2-thiouracil, 2 -thiothymine, 2-thiocytosine, 5-halouracil, cytosine, 5-propynyl uracil, 5-propynyl cytosine, 6-azo uracil, 6-azo cytosine, 6-azo thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-sulfhydryl, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo (e.g.. 5-bromo), 5-trifluoromethyl, and other 5-substituteduracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine, 7 -deazaadenine, 3 -deazaguanine, and 3 -deazaadenine.

[0069] In some embodiments, the 5’ and / or 3' end of the antisense strand can include abasic residues (Ab), which can also be referred to as an “abasic site” or “abasic nucleotide.” An abasic residue (Ab) is a nucleotide or nucleoside that lacks a nucleobase at the 1' position of the sugar moiety. (See, e.g., U.S. Patent No. 5,998.203). In some embodiments, an abasic residue can be placed internally in a nucleotide sequence. In some embodiments. Ab or AbAb can be added to the 3' end of the antisense strand. In some embodiments, the 5' end of the sense strand can include one or more additional abasic residues (e.g., (Ab) or (AbAb)). In some embodiments, UUAb, UAb, or Ab are added to the 3' end of the sense strand. In some embodiments, an abasic (deoxyribose) residue can be replaced with a ribitol (abasic ribose) residue.

[0070] In some embodiments, all or substantially all of the nucleotides of an RNAi agent are modified nucleotides. As used herein, an RNAi agent wherein substantially all of the nucleotides present are modified nucleotides is an RNAi agent having four or fewer (i.e., 0, 1, 2, 3, or 4) nucleotides in both the sense strand and the antisense strand being ribonucleotides (i.e., unmodified). As used herein, a sense strand wherein substantially all of the nucleotides present are modified nucleotides is a sense strand having two or fewer (i.e., 0, 1, or 2) nucleotides in the sense strand being unmodified ribonucleotides. As used herein, an antisense sense strand wherein substantially all of the nucleotides present are modified nucleotides is an antisense strand having two or fewer (i.e., 0, 1 , or 2) nucleotides in the sense strand being unmodified ribonucleotides. In some embodiments, one or more nucleotides of an RNAi agent is an unmodified ribonucleotide. Chemical structures for certain modified nucleotides are set forth in Table 1 1 herein.Modified Intemucleoside Linkages

[0071] In some embodiments, one or more nucleotides of an AR RNAi agent are linked by non-standard linkages or backbones (i.e., modified intemucleoside linkages or modified backbones). Modified intemucleoside linkages or backbones include, but are not limited to, phosphorothioate groups (represented herein as a lower case “s”). chiral phosphorothioates, thiophosphates, phosphorodithioates, phosphotri esters, aminoalkyl-phosphotriesters, alkyl phosphonates (e.g.. methyl phosphonates or 3'-alkylene phosphonates), chiral phosphonates, phosphinates, phosphoramidates (e.g., 3 '-amino phosphoramidate,annnoalkylphosphoramidates, or thionophosphoramidates), thionoalkyl-phosphonates, thionoalkylphosphotriesters, morpholino linkages, boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of boranophosphates, or boranophosphates having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'. In some embodiments, a modified intemucleoside linkage or backbone lacks a phosphorus atom. Modified intemucleoside linkages lacking a phosphorus atom include, but are not limited to, short chain alkyl or cycloalkyl inter-sugar linkages, mixed heteroatom and alkyl or cycloalkyl inter-sugar linkages, or one or more short chain heteroatomic or heterocyclic inter- sugar linkages. In some embodiments, modified intemucleoside backbones include, but are not limited to, siloxane backbones, sulfide backbones, sulfoxide backbones, sulfone backbones, formacetyl and thioformacetyl backbones, methylene formacetyl and thioformacetyl backbones, alkene-containing backbones, sulfamate backbones, methyleneimino and methylenehydrazino backbones, sulfonate and sulfonamide backbones, amide backbones, and other backbones having mixed N, O, S, and CH2 components.

[0072] In some embodiments, a sense strand of an AR RNAi agent can contain 1, 2, 3, 4. 5, or 6 phosphorothioate linkages, an antisense strand of an AR RNAi agent can contain 1, 2, 3, 4, 5, or 6 phosphorothioate linkages, or both the sense strand and the antisense strand independently can contain 1, 2, 3, 4, 5, or 6 phosphorothioate linkages. In some embodiments, a sense strand of an AR RNAi agent can contain 1, 2, 3, or 4 phosphorothioate linkages, an antisense strand of an AR RNAi agent can contain 1. 2, 3, or 4 phosphorothioate linkages, or both the sense strand and the antisense strand independently can contain 1 , 2, 3, or 4 phosphorothioate linkages.

[0073] In some embodiments, an AR RNAi agent sense strand contains at least two phosphorothioate intemucleoside linkages. In some embodiments, the phosphorothioate intemucleoside linkages are between the nucleotides at positions 1-3 from the 3’ end of the sense strand. In some embodiments, one phosphorothioate intemucleoside linkage is at the 5’ end of the sense strand nucleotide sequence, and another phosphorothioate linkage is at the 3’ end of the sense strand nucleotide sequence. In some embodiments, two phosphorothioate intemucleoside linkage are located at the 5’ end of the sense strand, and another phosphorothioate linkage is at the 3’ end of the sense strand. In some embodiments, the sense strand does not include any phosphorothioate intemucleoside linkages between the nucleotides, but contains one, two, or three phosphorothioate linkages between the terminal nucleotides on both the 5’ and 3’ ends and the optionally present inverted abasic residue terminal caps. Insome embodiments, the targeting ligand is linked to the sense strand via a phosphorothioate linkage.

[0074] In some embodiments, an AR RNAi agent antisense strand contains four phosphorothioate intemucleoside linkages. In some embodiments, the four phosphorothioate internucleoside linkages are between the nucleotides at positions 1-3 from the 5' end of the antisense strand and between the nucleotides at positions 19-21, 20-22, 21-23, 22-24, 23-25, or 24-26 from the 5' end. In some embodiments, three phosphorothioate intemucleoside linkages are located between positions 1-4 from the 5’ end of the antisense strand, and a fourth phosphorothioate intemucleoside linkage is located between positions 20-21 from the 5’ end of the antisense strand. In some embodiments, an AR RNAi agent contains at least three or four phosphorothioate intemucleoside linkages in the antisense strand.Capping Residues or Moieties

[0075] In some embodiments, the sense strand may include one or more capping residues or moieties, sometimes referred to in the art as a “cap,” a ‘■‘terminal cap,” or a “capping residue.” As used herein, a “capping residue” is a non-nucleotide compound or other moiety that can be incorporated at one or more termini of a nucleotide sequence of an RNAi agent disclosed herein. A capping residue can provide the RNAi agent, in some instances, with certain beneficial properties, such as, for example, protection against exonuclease degradation. In some embodiments, inverted abasic residues (invAb) (also referred to in the art as “inverted abasic sites”) are added as capping residues (see Table 1 1). (See, e.g., F. Czaudema. Nucleic Acids Res., 2003, 31(11), 2705-16). Capping residues are generally known in the art, and include, for example, inverted abasic residues as well as carbon chains such as a terminal C3H7 (propyl), C6H13 (hexyl), or C12H25 (dodecyl) groups. In some embodiments, a capping residue is present at either the 5' terminal end, the 3' terminal end, or both the 5' and 3' terminal ends of the sense strand. In some embodiments, the 5’ end and / or the 3' end of the sense strand may include more than one inverted abasic deoxyribose moiety as a capping residue.

[0076] In some embodiments, one or more inverted abasic residues (invAb) are added to the 3' end of the sense strand. In some embodiments, one or more inverted abasic residues (invAb) are added to the 5' end of the sense strand. In some embodiments, one or more inverted abasic residues or inverted abasic sites are inserted between the targeting ligand and the nucleotide sequence of the sense strand of the RNAi agent. In some embodiments, the inclusion of one or more inverted abasic residues or inverted abasic sites at or near the terminal end or terminalends of the sense strand of an RNAi agent allows for enhanced activity or other desired properties of an RN Ai agent.

[0077] In some embodiments, one or more inverted abasic residues (invAb) are added to the 3' end of the sense strand. In some embodiments, one or more inverted abasic residues (invAb) are added to the 5' end of the sense strand. In some embodiments, one or more inverted abasic residues or inverted abasic sites are inserted between the targeting ligand and the nucleotide sequence of the sense strand of the RNAi agent. In some embodiments, one or more inverted abasic residues or inverted abasic sites are inserted between the PK / PD modulator and the nucleotide sequence of the sense strand of the RNAi agent. In some embodiments, the inclusion of one or more inverted abasic residues or inverted abasic sites at or near the terminal end or terminal ends of the sense strand of an RNAi agent allows for enhanced activity or other desired properties of an RNAi agent.

[0078] In some embodiments, one or more inverted abasic residues (invAb) are added to the 5' end of the sense strand. In some embodiments, one or more inverted abasic residues can be inserted between the targeting ligand and the nucleotide sequence of the sense strand of the RNAi agent. The inverted abasic residues may be linked via phosphate, phosphorothioate (e.g.. shown herein as (invAb)s)), or other intemucleoside linkages. In some embodiments, the inclusion of one or more inverted abasic residues at or near the terminal end or terminal ends of the sense strand of an RNAi agent may allow for enhanced activity or other desired properties of an RNAi agent. In some embodiments, an inverted abasic (deoxyribose) residue can be replaced with an inverted ribitol (abasic ribose) residue. In some embodiments, the 3' end of the antisense strand core stretch sequence, or the 3' end of the antisense strand sequence, may include an inverted abasic residue. The chemical structures for inverted abasic deoxyribose residues are shown in Table 11 below.AR RNAi Agents

[0079] The AR RNAi agents disclosed herein are designed to target specific positions on an AR gene (e.g., SEQ ID NO: 1 (NM 000044.6)). As defined herein, an antisense strand sequence is designed to target an AR gene at a given position on the gene when the 5' terminal nucleobase of the antisense strand is aligned with a position that is 21 nucleotides downstream (towards the 3' end) from the position on the gene when base pairing to the gene. For example, as illustrated in Tables 1 and 2 herein, an antisense strand sequence designed to target an AR gene at position 304 requires that when base pairing to the gene, the 5' terminal nucleobase of the antisense strand is aligned with position 324 of an AR gene.

[0080] As provided herein, an AR RNAi agent does not require that the nucleobase at position 1 (5' -> 3') of the antisense strand be complementary to the gene, provided that there is at least 85% complementarity (e.g., at least 85, 86. 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% complementarity) of the antisense strand and the gene across a core stretch sequence of at least 16 consecutive nucleotides. For example, for an AR RNAi agent disclosed herein that is designed to target position 304 of an AR gene, the 5' terminal nucleobase of the antisense strand of the of the AR RNAi agent must be aligned with position 324 of the gene; however, the 5' terminal nucleobase of the antisense strand may be, but is not required to be, complementary to position 324 of an AR gene, provided that there is at least 85% complementarity (e.g., at least 85, 86, 87, 88, 89, 90. 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% complementarity) of the antisense strand and the gene transcript across a core stretch sequence of at least 16 consecutive nucleotides. As shown by, among other things, the various examples disclosed herein, the specific site of binding of the gene by the antisense strand of the AR RNAi agent (e.g., whether the AR RNAi agent is designed to target an AR gene at position 127, at position 130. at position 136, or at some other position) is an important factor to the level of inhibition achieved by the AR RNAi agent. (See, e.g., Kamola et al., The siRNA Non-seed Region and Its Target Sequences are Auxiliary Determinants of Off-Target Effects, PLOS Computational Biology, 11(12), Figure 1 (2015)).

[0081] In some embodiments, the AR RNAi agents disclosed herein target an AR gene al or near the positions of the AR sequence shown in Table 1 . In some embodiments, the antisense strand of an AR RNAi agent disclosed herein includes a core stretch sequence that is fully, substantially, or at least partially complementary to a target AR 19-mer sequence disclosed in Table 1.Table 1. AR 19-mer mRNA Target Sequences (taken from homo sapiens Androgen Receptor(AR) transcript, GenBank NM_000044.6 (SEQ ID NO: 1))

[0082] Homo sapiens Androgen Receptor (AR), GenBank NM_000044.6 (SEQ ID NO: 1), gene transcript (10667 bases):1 agcgccccct ccgagatccc ggggagccag cttgctggga gagcgggacg gtccggagca61 agcccagagg cagaggaggc gacagaggga aaaagggccg agctagccgc tccagtgctg121 tacaggagcc gaagggacgc accacgccag ccccagcccg gctccagcga cagccaacgc181 ctcttgcagc gcggcggctt cgaagccgcc gcccggagct gcccttcct cttcggtgaa241 gttttaaaa gctgctaaag actcggagga agcaaggaaa gtgcctggta ggactgacgg301 ctgcctttgt cctcctcctc tccaccccgc ctccccccac cctgccttcc ccccctcccc361 cgtcttctct cccgcagctg cctcagtcgg ctactctcag ccaacccccc tcaccaccct421 tctccccacc cgcccccccg cccccgtcgg cccagcgctg ccagcccgag tttgcagaga481 ggtaactccc tttggctgcg agcgggcgag ctagctgcac attgcaaaga aggctcttag541 gagccaggcg actggggagc ggcttcagca ctgcagccac gacccgcctg gtaggctgc601 acgcggagag aaccctctgt tttcccccac tctctctcca cctcctcctg ccttccccac661 cccgagtgcg gagccagaga tcaaaagatg aaaaggcagt caggtcttca gtagccaaaa 721 aacaaaacaa acaaaaacaa aaaagccgaa ataaaagaaa aagataataa ctcagttctt 781 atttgcacct acttcagtgg acactgaatt tggaaggtgg aggattttgt ttttttcttt841 taagatctgg gcatcttttg aatctaccct tcaagtatta agagacagac tgtgagccta 901 gcagggcaga tcttgtccac cgtgtgtctt cttctgcacg agactttgag gctgtcagag 961 cgctttttgc gtggttgctc ccgcaagttt ccttctctgg agcttcccgc aggtgggcag 1021 ctagctgcag cgactaccgc atcatcacag cctgttgaac tcttctgagc aagagaaggg 1081 gaggcggggt aagggaagta ggtggaagat tcagccaagc tcaaggatgg aagtgcagtt 1141 agggctggga agggtctacc ctcggccgcc gtccaagacc taccgaggag ctttccagaa 1201 tctgttccag agcgtgcgcg aagtgatcca gaacccgggc cccaggcacc cagaggccgc 1261 gagcgcagca cctcccggcg ccagtttgct gctgctgcag cagcagcagc agcagcagca 1321 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcaagaga ctagccccag 1381 gcagcagcag cagcagcagg gtgaggatgg ttctccccaa gcccatcgta gaggccccac 1441 aggctacctg gtcctggatg aggaacagca accttcacag ccgcagtcgg ccctggagtg 1501 ccaccccgag agaggttgcg tcccagagcc tggagccgcc gtggccgcca gcaaggggct 1561 gccgcagcag ctgccagcac ctccggacga ggatgactca gctgccccat ccacgttgtc 1621 cctgctgggc cccacttcc ccggcttaag cagctgctcc gctgacctta aagacatcct 1681 gagcgaggcc agcaccatgc aactccttca gcaacagcag caggaagcag tatccgaagg 1741 cagcagcagc gggagagcga gggaggcctc gggggctccc actcctcca aggacaatta 1801 cttagggggc acttcgacca tttctgacaa cgccaaggag ttgtgtaagg cagtgtcggt 1861 gtccatgggc ctgggtgtgg aggcgttgga gcatctgagt ccaggggaac agcttcgggg 1921 ggattgcatg tacgccccac ttttgggagt tccacccgct gtgcgtccca ctccttgtgc 1981 cccattggcc gaatgcaaag gttctctgct agacgacagc gcaggcaaga gcactgaaga 2041 tactgctgag tattcccctt tcaagggagg ttacaccaaa gggctagaag gcgagagcct 2101 aggctgctct ggcagcgctg cagcagggag ctccgggaca cttgaactgc cglctaccct 2161 gtctctctac aagtccggag cactggacga ggcagctgcg taccagagtc gcgactacta 2221 caactttcca ctggctctgg ccggaccgcc gccccctccg ccgcctcccc atccccacgc 2281 tcgcatcaag ctggagaacc cgctggacta cggcagcgcc tgggcggctg cggcggcgca 2341 gtgccgctat ggggacctgg cgagcctgca tggcgcgggt gcagcgggac ccggttctgg 2401 gtcaccctca gccgccgctt cctcatcctg gcacactctc ttcacagccg aagaaggcca 2461 gtgtatgga ccgtgtggtg gtggtggggg tggtggcggc ggcggcggcg gcggcggcgg 2521 cggcggcggc ggcggcggcg gcggcgaggc gggagctgta gccccctacg gctacactcg 2581 gccccctcag gggctggcgg gccaggaaag cgacttcacc gcacctgatg tgtggtaccc2641 tggcggcatg gtgagcagag tgccctatcc cagtcccact tgtgtcaaaa gcgaaatggg2701 cccctggatg gatagctact ccggacctta cggggacatg cgtttggaga ctgccaggga2761 ccatgttttg cccattgact attactttcc accccagaag acctgcctga tctgtggaga2821 tgaagcttct gggtgtcact atggagctct cacatgtgga agctgcaagg tcttcttcaa2881 aagagccgct gaagggaaac agaagtacct gtgcgccagc agaaatgatt gcactattga2941 taaattccga aggaaaaatt gtccatcttg tcgtcttcgg aaatgttatg aagcagggat3001 gactctggga gcccggaagc tgaagaaact tggtaatctg aaactacagg aggaaggaga3061 ggcttccagc accaccagcc ccactgagga gacaacccag aagctgacag tgtcacacat3121 tgaaggclat gaatgtcagc ccatclttct gaatgtcctg gaagccatg agccaggtgt3181 agtgtgtgct ggacacgaca acaaccagcc cgactccttt gcagcctgc tctctagcct3241 caatgaactg ggagagagac agcttgtaca cgtggtcaag tgggccaagg ccttgcctgg3301 cttccgcaac ttacacgtgg acgaccagat ggctgtcatt cagtactcct ggatggggct3361 catggtgttt gccatgggct ggcgatcctt caccaatgtc aactccagga tgctctactt3421 cgcccctgat ctggttttca atgagtaccg catgcacaag tcccggatgt acagccagtg3481 tgtccgaatg aggcacctct ctcaagagtt tggatggctc caaatcaccc cccaggaatt3541 cctgtgcatg aaagcactgc tactcttcag cattattcca gtggatgggc tgaaaaatca3601 aaaatcttt gatgaacttc gaatgaacta catcaaggaa ctcgatcgta tcattgcatg3661 caaaagaaaa aatcccacat cctgctcaag acgcttctac cagctcacca agctcctgga3721 ctccglgcag cctattgcga gagagctgca tcagttcact tttgacctgc taatcaagtc3781 acacatggtg agcgtggact ttccggaaat gatggcagag atcatctctg tgcaagtgcc3841 caagatcctt tctgggaaag tcaagcccat ctatttccac acccagtgaa gcattggaaa3901 ccctatttcc ccaccccagc tcatgccccc tttcagatgt cttctgcctg ttataactct3961 gcactactcc tctgcagtgc cttggggaat ttcctctatt gatgtacagt ctgtcatgaa4021 catgttcctg aattctattt gctgggcttt ttttt tctct ttctctcctt tcttttct4081 cttccctccc tatctaaccc tcccatggca ccttcagact ttgcttccca ttgtggctcc4141 tatctgtgtt ttgaatggtg ttgtatgcct ttaaatctgt gatgatcctc atatggccca4201 gtgtcaagtt gtgcttgtt acagcactac tctgtgccag ccacacaaac gtttacttat4261 cttatgccac gggaagttta gagagctaag attatctggg gaaatcaaaa caaaaacaag4321 caaacaaaaa aaaaaagcaa aaacaaaaca aaaaataagc caaaaaacct tgctagtgtt4381 tttcctcaa aaataaataa ataaataaat aaatacgtac atacatacac acatacatac4441 aaacatatag aaatccccaa agaggccaat agtgacgaga aggtgaaaat tgcaggccca4501 tggggagtta ctgatttttt catctcctcc ctccacggga gactttattt tctgccaatg4561 gctattgcca ttagagggca gagtgacccc agagctgagt tgggcagggg ggtggacaga4621 gaggagagga caaggagggc aatggagcat cagtacctgc ccacagcctt ggtccctggg4681 ggctagactg ctcaactgtg gagcaattca ttatactgaa aatgtgcttg tgttgaaaa4741 tttgtctgca tgttaatgcc tcacccccaa acccttttct ctctcactct ctgcctccaa4801 cttcagattg actttcaata gtttttctaa gacctttgaa ctgaatgttc tcttcagcca4861 aaacttggcg acttccacag aaaagtctga ccactgagaa gaaggagagc agagatttaa4921 ccctttgtaa ggccccattt ggatccaggt ctgctttctc atgtgtgagt cagggaggag4981 ctggagccag aggagaagaa aatgatagct tggctgttct cctgcttagg acactgactg5041 aatagttaaa ctctcactgc cactaccttt tccccacctt taaaagacct gaatgaagtt5101 ttctgccaaa ctccgtgaag ccacaagcac ctatgtcct cccttcagtg ttttgtgggc5161 ctgaatttca tcacactgca tttcagccat ggtcatcaag cctgtttgct tctttgggc5221 atgttcacag attctctgtt aagagccccc accaccaaga aggttagcag gccaacagct5281 ctgacatcta tctgtagatg ccagtagtca caaagatttc ttaccaactc tcagatcgct5341 ggagccctta gacaaactgg aaagaaggca tcaaagggat caggcaagct gggcgtctg5401 ccctgtccc ccagagatga taccctccca gcaagtggag aagttctcac ttcctcttt5461 agagcagcta aaggggctac ccagatcagg gttgaagaga aaactcaatt accagggtgg5521 gaagaatgaa ggcactagaa ccagaaaccc tgcaaatgct cttcttgtca cccagcatat5581 ccacctgcag aagtcatgag aagagagaag gaacaaagag gagactctga ctactgaatt5641 aaaatcttca gcggcaaagc ctaaagccag atggacacca tctggtgagt ttactcatca5701 tcctcctctg ctgctgattc tgggctctga cattgcccat actcactcag attccccacc5761 tttgttgctg cctcltagtc agagggaggc caaaccallg agactttcta cagaaccatg5821 gcttctttcg gaaaggtctg gttggtgtgg ctccaatact ttgccaccca tgaactcagg5881 gtgtgccctg ggacactggt tttatatagt cttttggcac acctgtgttc tgttgacttc5941 gttcttcaag cccaagtgca agggaaaatg tccacctact ttctcatctt ggcctctgcc6001 tccttactta gctcttaatc tcatctgtg aactcaagaa atcaagggcc agtcatcaag6061 ctgcccattt taattgattc actctgtttg tgagaggat agtttctgag tgacatgata6121 tgatccacaa gggtttcctt ccctgatttc tgcattgata ttaatagcca aacgaacttc6181 aaaacagctt taaataacaa gggagagggg aacctaagat gagtaatatg ccaatccaag6241 actgctggag aaaactaaag ctgacaggtt ccctttttgg ggtgggatag acatgtctg6301 gttttctta ttattacaca atctggctca tgtacaggat cacttttagc tgttttaaac6361 agaaaaaaat atccaccact cttttcagtt acactaggtt acattttaat aggtccttta6421 catctgttt ggaatgatt tcatcttttg tgatacacag attgaattat atcattttca6481 tatctctcct tgtaaatact agaagctctc ctttacatt ctctatcaaa tttttcatct6541 ttatgggttt cccaattgtg actcttgtct tcatgaatat atgtttttca tttgcaaaag6601 ccaaaaatca gtgaaacagc agtgtaatta aaagcaacaa ctggatlact ccaaatttcc6661 aaatgacaaa actagggaaa aatagcctac acaagccttt aggcctactc tttctgtgct6721 tgggtttgag tgaacaaagg agattttagc ttggctctgt tctcccatgg atgaaaggag6781 gaggattttt tttttctttt ggccattgat gttctagcca atgtaattga cagaagtctc6841 attttgcatg cgctctgctc tacaaacaga gttggtatgg ttggtatact gtactcacct6901 gtgagggact ggccactcag acccacttag ctggtgagct agaagatgag gatcactcac6961 tggaaaagtc acaaggacca tctccaaaca agttggcagt gctcgatgtg gacgaagagt7021 gaggaagaga aaaagaagga gcaccaggga gaaggctccg tctgtgctgg gcagcagaca7081 gctgccagga tcacgaactc tgtagtcaaa gaaaagagtc gtgtggcagt ttcagctctc7141 gttcattggg cagctcgcct aggcccagcc tctgagctga catgggagtt gttggattct7201 ttgtttcata gctttttcta tgccataggc aatattgttg ttcttggaaa gtttattatt7261 tttttaactc ccttactctg agaaagggat attttgaagg actgtcatat atctttgaaa7321 aaagaaaatc tgtaatacat atatttttat gtatgttcac tggcactaaa aaatatagag7381 agcttcattc tgtcctttgg gtagttgctg aggtaattgt ccaggttgaa aaataatgtg7441 ctgatgctag agtccctctc tgtccatact ctacttctaa atacatatag gcatacatag7501 caagttttat ttgacttgta ctttaagaga aaatatgtcc accatccaca tgatgcacaa7561 atgagctaac attgagcttc aagtagcttc taagtgtttg tttcattagg cacagcacag7621 atgtggcctt tccccccttc tctcccttga tatctggcag ggcataaagg cccaggccac7681 ttcctctgcc ccttcccagc cctgcaccaa agctgcattt caggagactc tctccagaca7741 gcccagtaac tacccgagca tggcccctgc atagccctgg aaaaataaga ggctgactgt7801 ctacgaatla tcttgtgcca gttgcccagg tgagagggca ctgggccaag ggagtggttl7861 tcatgtttga cccactacaa ggggtcatgg gaatcaggaa tgccaaagca ccagatcaaa7921 tccaaaactt aaagtcaaaa taagccattc agcatgttca gtttcttgga aaaggaagtt7981 tctacccctg atgcctttgt aggcagatct gttctcacca ttaatctttt tgaaaatctt8041 ttaaagcagt ttttaaaaag agagatgaaa gcatcacatt atataaccaa agattacatt8101 gtacctgcta agataccaaa attcataagg gcaggggggg agcaagcatt agtgcctctt8161 tgataagctg tccaaagaca gactaaagga ctctgctggt gactgactta taagagcttt8221 gtgggttttt tttlccctaa taatatacat gtttagaaga attgaaaata atttcgggaa8281 aatgggatta tgggtccttc actaagtgat tttataagca gaactggctt tccttttctc8341 tagtagttgc tgagcaaatt gttgaagctc catcattgca tggttggaaa tggagctgtt8401 cttagccact gtgtttgcta gtgcccatgt tagcttatct gaagatgtga aacccttgct8461 gataagggag catttaaagt actagatttt gcactagagg gacagcaggc agaaatcctt8521 atttctgccc actttggatg gcacaaaaag ttatctgcag ttgaaggcag aaagttgaaa8581 tacattgtaa atgaatattt gtatccatgt ttcaaaattg aaatatatat atatatatat8641 atatatatat atatatatat atagtgtgtg tgtgtgttct gatagcttta actttctctg8701 catctttata tttggttcca gatcacacct gatgccatgt acttgtgaga gaggatgcag8761 ttttgttttg gaagctctct cagaacaaac aagacacctg gattgatcag ttaactaaaa 8821 gttttctccc ctattgggtt tgacccacag gtcctgtgaa ggagcagagg gataaaaaga 8881 gtagaggaca tgatacattg tactttacta gttcaagaca gatgaatgtg gaaagcataa 8941 aaactcaatg gaactgactg agatttacca cagggaaggc ccaaacttgg ggccaaaagc 9001 ctacccaagt gattgaccag tggcccccta atgggacctg agctgttgga agaagagaac 9061 tgttccttgg tcttcaccat cctgtgaga gaagggcagt ttcctgcatt ggaacctgga 9121 gcaagcgctc tatctttcac acaaattccc tcacctgaga ttgaggtgct cttgttactg 9181 ggtgtctgtg tgctgtaatt ctggttttgg atatgttctg taaagatttt gacaaatgaa 9241 aatgtgtttt tctctgttaa aacttgtcag agtactagaa gttgtatctc tgtaggtgca 9301 ggtccatttc tgcccacagg tagggtgttt ttctttgatt aagagattga cacttctgtt 9361 gcctaggacc tcccaactca accatttcta ggtgaaggca gaaaaatcca cattagttac 9421 tcctcttcag acatttcagc tgagataaca aatcttttgg aatttttca cccatagaaa 9481 gagtggtaga tatttgaatt tagcaggtgg agtttcatag taaaaacagc ttttgactca 9541 gctttgattt atcctcattt gatttggcca gaaagtaggt aatatgcatt gatggcttc 9601 tgattccaat tcagtatagc aaggtgctag gttttttcct ttccccacct gtctctagc 9661 ctggggaatt aaatgagaag ccttagaatg ggtggccctt gtgacctgaa acacttccca 9721 cataagctac ttaacaagat tgtcatggag ctgcagattc cattgcccac caaagactag 9781 aacacacaca tatccataca ccaaaggaaa gacaattctg aaatgctgtt tctctggtgg 9841 ttccclctct ggctgctgcc tcacagtatg ggaacclgta ctctgcagag gtgacaggcc 9901 agatttgcat tatctcacaa ccttagccct tggtgctaac tgtcctacag tgaagtgcct 9961 ggggggttgt cctatcccat aagccacttg gatgctgaca gcagccacca tcagaatgac 10021 ccacgcaaaa aaaagaaaaa aaaaattaaa aagtcccctc acaacccagt gacacctttc 10081 tgctttcctc tagactggaa cattgattag ggagtgcctc agacatgaca ttcttgtgct 10141 gtcctggaa taatctggc agcaggaggg agcagactat gtaaacagag ataaaaatta 10201 attttcaata ttgaaggaaa aaagaaataa gaagagagag agaaagaaag catcacacaa 10261 agattttct aaaagaaaca attttgcttg aaatctcttt agatggggct catttctcac 10321 ggtggcactt ggcctccact gggcagcagg accagctcca agcgctagtg ttctgtctc 10381 tttttgtaat cttggaatct tttgttgctc taaatacaat taaaaatggc agaaacttgt 10441 ttgttggact acatgtgtga ctttgggtct gtctctgcct ctgctttcag aaatgtcatc 10501 cattgtgtaa aatatggct tactggtctg ccagctaaaa cttggccaca tcccctgtla 10561 tggctgcagg atcgagttat tgttaacaaa gagacccaag aaaagctgct aatgtcctct 10621 tatcattgtt gttaatttgt taaaacataa agaaatctaa aatttca

[0083] In some embodiments, an AR RNAi agent includes an antisense strand wherein position 19 of the antisense strand (5 ' 3 ') is capable of forming a base pair with position 1 of a 19-mer target sequence disclosed in Table 1. In some embodiments, an AR agent includes an antisense strand wherein position 1 of the antisense strand (5 ' — > 3 ') is capable of forming a base pair with position 19 of a 19-mer target sequence disclosed in Table 1.

[0084] In some embodiments, an AR agent includes an antisense strand wherein position 2 of the antisense strand (5'3’) is capable of forming a base pair with position 18 of a 19-mer target sequence disclosed in Table 1 . In some embodiments, an AR agent includes an antisense strand wherein positions 2 through 18 of the antisense strand (5' -> 3') are capable of forming base pairs with each of the respective complementary bases located at positions 18 through 2 of the 19-mer target sequence disclosed in Table 1.

[0085] For the RNAi agents disclosed herein, the nucleotide at position 1 of the antisense strand (from 5' end -> 3' end) can be perfectly complementary to an AR gene, or can be non- complementary to an AR gene. In some embodiments, the nucleotide at position 1 of the antisense strand (from 5' end3' end) is a U, A, or dT. In some embodiments, the nucleotide at position 1 of the antisense strand (from 5' end -> 3' end) forms an A:U or U:A base pair with the sense strand.

[0086] In some embodiments, an AR RNAi agent antisense strand comprises the sequence of nucleotides (from 5' end -> 3' end) 2-18 or 2-19 of any of the antisense strand sequences in Table 2 or Table 3 In some embodiments, an AR RNAi sense strand comprises the sequence of nucleotides (from 5' end -> 3' end) 1 -17, 1 -18, or 2-18 of any of the sense strand sequences in Table 2, Table 4, Table 5, or Table 6.

[0087] In some embodiments, an AR RNAi agent comprises of (i) an antisense strand comprising the sequence of nucleotides (from 5' end -> 3' end) 2-18 or 2-19 of any of the antisense strand sequences in Table 2 or Table 3, and (li) a sense strand comprising the sequence of nucleotides (from 5' end3' end) 1-17 or 1-18 of any of the sense strand sequences in Table 2, Table 4, Table 5, or Table 6.

[0088] In some embodiments, the AR RNAi agents include core 19-mer nucleotide sequences shown in the following Table 2.Table 2. AR RNAi Agent Antisense Strand and Sense Strand Core Stretch Base Sequences (N=any nucleobase; I = inosine (hypoxanthine nucleobase)

[0089] The AR RNAi agent sense strands and antisense strands that comprise or consist of the nucleotide sequences in Table 2 can be modified nucleotides or unmodified nucleotides. In some embodiments, the AR RNAi agents having the sense and antisense strand sequences that comprise or consist of any of the nucleotide sequences in Table 2 are all or substantially all modified nucleotides.

[0090] In some embodiments, the antisense strand of an AR RNAi agent disclosed herein differs by 0, 1, 2, or 3 nucleotides from any of the antisense strand sequences in Table 2. In some embodiments, the sense strand of an AR RNAi agent disclosed herein differs by 0, 1, 2, or 3 nucleotides from any of the sense strand sequences in Table 2.

[0091] As used herein, each N listed in a sequence disclosed in Table 2 may be independently selected from any and all nucleobases (including those found on both modified and unmodified nucleotides). In some embodiments, an N nucleotide listed in a sequence disclosed in Table 2 has a nucleobase that is complementary to the N nucleotide at the corresponding position on the other strand. In some embodiments, an N nucleotide listed in a sequence disclosed in Table 2 has a nucleobase that is not complementary to the N nucleotide at the corresponding position on the other strand. In some embodiments, an N nucleotide listed in a sequence disclosed in Table 2 has a nucleobase that is the same as the N nucleotide at the corresponding position on the other strand. In some embodiments, an N nucleotide listed in a sequence disclosed in Table 2 has a nucleobase that is different from the N nucleotide al the corresponding position on the other strand.

[0092] Certain modified AR RNAi agent sense and antisense strands are provided in Table 3, Table 4, Table 5, Table 6, and Table 10. Certain modified AR RNAi agent antisense strands, as well as their underlying unmodified nucleobase sequences, are provided in Table 3. Certain modified AR RNAi agent sense strands, as well as their underlying unmodified nucleobase sequences, are provided in Tables 4. 5, and 6. In forming AR RNAi agents, each of the nucleotides in each of the underlying base sequences listed in Tables 3, 4, 5, and 6, as well as in Table 2, above, can be a modified nucleotide.

[0093] The AR RNAi agents described herein are formed by annealing an antisense strand with a sense strand. A sense strand containing a sequence listed in Table 2, Table 4, Table 5, or Table 6 can be hybridized to any antisense strand containing a sequence listed in Table 2 or Table 3, provided the two sequences have a region of at least 85% complementarity over a contiguous 16, 17, 18, 19, 20, or 21 nucleotide sequence.

[0094] In some embodiments, an AR RNAi agent antisense strand comprises a nucleotide sequence of any of the sequences in Table 2 or Table 3.

[0095] In some embodiments, an AR RNAi agent comprises or consists of a duplex having the nucleobase sequences of the sense strand and the antisense strand of any of the sequences in Table 2, Table 3, Table 4, Table 5, Table 6, or Table 10.

[0096] Examples of antisense strands containing modified nucleotides are provided in Table 3. Examples of sense strands containing modified nucleotides are provided in Tables 4, 5 and 6.

[0097] As used in Tables 3, 4. 5, 6. and 10, the following notations are used to indicate modified nucleotides, targeting groups, and linking groups:A = adenosine-3'-phosphateC = cytidine-3 '-phosphateG = guanosine-3'-phosphateU = uridine-3 '-phosphateI = inosine-3 '-phosphate a = 2'-O-methyladenosine-3 '-phosphate as = 2'-O-methyladenosine-3'-phosphorothioate c = 2'-O-methylcytidine-3 '-phosphate cs = 2'-O-methylcytidine-3'-phosphorothioate g = 2'-O-methylguanosine-3'-phosphate gs = 2'-O-methylguanosine-3'-phosphorothioate i = 2'-O-methylinosine-3 ’-phosphate is = 2'-O-methylinosine-3’-phosphorothioate t = 2'-O-methyl-5-methyluridine-3 '-phosphate ts = 2'-O-methyl-5-methyluridine-3'-phosphorothioate u = 2'-O-methyluridine-3 '-phosphate us = 2'-O-methyluridine-3'-phosphorothioateAf = 2'-fluoroadenosine-3'-phosphateAfs = 2'-fluoroadenosine-3'-phosporothioateCf = 2'-fluorocytidine-3 '-phosphateCfs = 2'-fluorocytidine-3'-phosphorothioateGf = 2'-fluoroguanosine-3 '-phosphateGfs = 2'-fluoroguanosine-3'-phosphorothioateTf = 2'-fluoro-5'-methyluridine-3'-phosphateTfs = 2'-fluoro-5'-methyluridine-3'-phosphorothioateUf = 2'-fluorouridine-3 '-phosphateUfs = 2'-fluorouridine-3'-phosphorothioate dT = 2'-deoxythymidine-3 '-phosphateAUNA = 2'.3'-seco-adenosine-3'-phosphate (See Table 11)AUNAS = 2'.3'-seco-adenosine-3'-phosphorothioate (See Table 11) CUNA = 2',3'-seco-cytidine-3 '-phosphate (See Table 11)CUNAS = 2'.3'-seco-cytidine-3'-phosphorothioate (See Table 11)GUNA = 2',3'-seco-guanosine-3 '-phosphate (See Table 11)GUNAS = 2',3'-seco-guanosine-3'-phosphorothioate (See Table 11)UUNA = 2',3'-seco-uridine-3'-phosphate (See Table 11)UUNAS = 2',3'-seco-uridine-3'-phosphorothioate (See Table 11 ) a_2N = see Table 11 a_2Ns = see Table 11(invAb) = inverted abasic deoxyribonucleotide-5'- phosphate, see Table 11(invAb)s = inverted abasic deoxyribonucleotide-5'- phosphorothioate, see Table 11 s = phosphorothioate linkage p = terminal phosphate (as synthesized) vpdN = vinyl phosphonate deoxyribonucleotide cPrpa = 5 ’-cyclopropyl phosphonate-2'-O-methyladenosine-3'-phosphate(see Table 11) cPrpas = 5 ’-cyclopropyl phosphonate-2'-O-methyladenosine-3'- phosphorothioate (see Table 11) cPrpu = 5 ’-cyclopropyl phosphonate-2'-O-methyluridine-3'-phosphate (seeTable 11) cPrpus = 5 ’-cyclopropyl phosphonate-2'-O-methyluridine-3'- phosphorothioate (see Table 11)(Alk-SS-C6) = see Table 11(C6-SS-Alk) = see Table 11(C6-SS-C6) = see Table 11(6-SS-6) = see Table 11(C6-SS-Alk-Me) = see Table 11(NH2-C6) = see Table 11-C6- = see Table 11-C6s- = see Table 11-L6-C6- = see Table 11-L6-C6s- = see Table 11LP183s = see Table l lLP183rs = see Table l lLP409s = see Table 11 cC16 = see Table l l aC16 = see Table l l gC16 = see Table l l uC16 = see Table l lALNA = see Table 11 cl 6s = see Table l lC22s = see Table 11H0-C16s = see Table 11(2C8C12)s = see Table 11(2C6C10)s = see Table 11LP283 = see Table 11LP293 = see Table 11LP294 = see Table 11LP310 = seeTable llLP383 = seeTablellLP395 = seeTablellLP395s = seeTable llLP396 = seeTable ll aAlk = see Table 11 uAlk = seeTable ll cAlk = seeTablell gAlk = seeTablell(NAG37) = seeTable ll(NAG37)s = see Table 11[NEM] = seeTable llFabOOOl = see Antigen Binding Proteins, infraFab0002 = see Antigen Binding Proteins, infraFab0044 = see Antigen Binding Proteins, infraFab0046 = see Antigen Binding Proteins, infraFab0056 = see Antigen Binding Proteins, infraFab0058 = see Antigen Binding Proteins, infraFab0060 = see Antigen Binding Proteins, infraFab0061 = see Antigen Binding Proteins, infraFab0062 = see Antigen Binding Proteins, infraFab0063 = see Antigen Binding Proteins, infraFab0064 = see Antigen Binding Proteins, infraFab0065 = see Antigen Binding Proteins, infraFab0066 = see Antigen Binding Proteins, infraFab0067 = see Antigen Binding Proteins, infraFab0068 = see Antigen Binding Proteins, infraFab0069 = see Antigen Binding Proteins, infraFab0070 = see Antigen Binding Proteins, infraFab0071 = see Antigen Binding Proteins, infraFab0072 = see Antigen Binding Proteins, infraFab0073 = see Antigen Binding Proteins, infraFab0074 = see Antigen Binding Proteins, infraFab0165 = see Antigen Binding Proteins, infraFab0166 = see Antigen Binding Proteins, infraFab0167 = see Antigen Binding Proteins, infraFab0168 = see Antigen Binding Proteins, infra xAb000293 = see Antigen Binding Proteins, infra LP-29 = see Table 1 1LP-238b = see Table 11 av[36-pepl = see Table 11, also referred to as "avp6 peptide 1”

[0098] As the person of ordinary skill in the art would readily understand, unless otherwise indicated by the sequence (such as, for example, by a phosphorothioate linkage “s”), when present in an oligonucleotide, the nucleotide monomers are mutually linked by 5’-3’- phosphodiester bonds. As the person of ordinary skill in the art would dearly understand, the inclusion of a phosphorothioate linkage as shown in the modified nucleotide sequences disclosed herein replaces the phosphodiester linkage typically present in oligonucleotides. Further, the person of ordinary skill in the art would readily understand that the terminal nucleotide at the 3’ end of a given oligonucleotide sequence would typically have a hydroxyl (-OH) group at the respective 3’ position of the given monomer instead of a phosphate moiety ex vivo. Additionally, for the embodiments disclosed herein, when viewing the respective strand 5’ -> 3’, the inverted abasic residues are inserted such that the 3’ position of the deoxyribose is linked at the 3’ end of the preceding monomer on the respective strand (see, e.g., Table 11). Moreover, as the person of ordinary skill would readily understand and appreciate, while the phosphorothioate chemical structures depicted herein typically show the anion on the sulfur atom, the inventions disclosed herein encompass all phosphorothioate tautomers (e.g., where the sulfur atom has a double-bond and the anion is on an oxygen atom). Unless expressly indicated otherwise herein, such understandings of the person of ordinary skill in the art are used when describing the AR RNAi agents and compositions of AR RNAi agents disclosed herein.

[0099] Certain examples of targeting groups and linking groups used with the AR RN Ai agents disclosed herein are included in the chemical structures provided below in Table 11. Each sense strand and / or antisense strand can have any targeting groups or linking groups listed herein, as well as other targeting or linking groups, conjugated to the 5' and / or 3' end of the sequence.

[0100] Certain examples of antigen binding proteins and linking groups used with the AR RNAi agents disclosed herein are included in the chemical structures provided below in Table 1 1 . Each sense strand and / or antisense strand can have any antigen binding protein or linkinggroup listed herein, as well as other targeting groups, antigen binding proteins, linking groups, conjugated to the 5' and / or 3' end of the sequenceTable 3. AR RNAi Agent Antisense Strand SequencesTable 4. AR Agent Sense Strand Sequences (Shown Without Linkers, Conjugates, or Capping Moieties)a_2N=2-aminoadenosine nucleotide, (A2N) = 2-aminoadenine nucleotideTable 5. AR Agent Sense Strand Sequences (Shown With (NH2-C6) Linker or (NAG37)s ligand (see Table 1 1 for structure information.)) a 2N=2-aminoadenosine nucleotidea_2N=2-aminoadenosine nucleotide, (A2N) = 2-aminoadenine nucleotideTable 6. AR Agent Sense Strand Sequences (Shown with lipid moiety or antigen binding moiety). The structures of the lipid moieties are shown in Table 11.Table 6.1. AR RNAi Agent Sense Strand Sequences Showing Targeting Ligand and / or PK / PD modulator Positions (Z = pharmacological moiety (e.g., targeting ligand, targeting group, and / or PK / PD modulator)). The structures of the lipid moieties are shown in Table 11.Table 6.2. AR RNAi Agent Sense Strand Sequences Showing Targeting Ligand linked at the 5’ terminal end and PK / PD modulator linked at the 3’ terminal end of the sense strand. (TL = targeting ligand; PK = PK / PD modulator)).

[0101] The AR RNAi agents disclosed herein are formed by annealing an antisense strand with a sense strand. A sense strand containing a sequence listed in Table 2. Table 4, Table 5. or Table 6 can be hybridized to any antisense strand containing a sequence listed in Table 2 or Table 3, provided the two sequences have a region of at least 85% complementarity over a contiguous 16, 17, 18, 19, 20, or 21 nucleotide sequence.

[0102] As shown in Table 5 above, certain of the example AR RNAi agent nucleotide sequences are shown to further include reactive linking groups at one or both of the 5’ terminal end and the 3’ terminal end of the sense strand. For example, many of the AR RNAi agent sense strand sequences shown in Table 5 above have a (NH2-C6) linking group at the 5’ end of the nucleotide sequence. Other linking groups, such as a (6-SS-6) linking group or a (C6-SS-C6) linking group, may be present as w ell or alternatively in certain embodiments. Such reactive linking groups are positioned to facilitate the linking of targeting ligands, targeting groups, and / or PK / PD modulators to the AR RNAi agents disclosed herein. Linking or conjugation reactions are well known in the art and provide for formation of covalent linkages between two molecules or reactants. Suitable conjugation reactions for use in the scope of the inventions herein include, but are not limited to. amide coupling reaction, Michael addition reaction, hydrazone formation reaction, inverse-demand Diels-Alder cycloaddition reaction, oxime ligation, and Copper (I)- catalyzed or strain-promoted azide-alkyne cycloaddition reaction cycloaddition reaction.

[0103] As shown in Table 5 above, certain of the example AR RNAi agent nucleotide sequences are show n to further include reactive linking groups at one or both of the 5’ terminal end and the 3’ terminal end of the sense strand. For example, many of the AR RNAi agent sense strand sequences shown in Table 5 above have a (NH2-C6) linking group at the 5’ end of the nucleotide sequence. Other linking groups, such as a (6-SS-6) linking group or a (C6-SS-C6) linking group, may be present as well or alternatively in certain embodiments. Such reactive linking groups are positioned to facilitate the linking of targeting ligands, targeting groups, and / or antigen binding proteins to the AR RNAi agents disclosed herein. Linking or conjugation reactions are well known in the art and provide for formation of covalent linkages between two molecules or reactants. Suitable conjugation reactions for use in the scope of the inventions herein include, but are not limited to, amide coupling reaction, Michael addition reaction, hydrazone formation reaction, inverse-demand Diels-Alder cycloaddition reaction, oxime ligation, and Copper (I)- catalyzed or strain-promoted azide-alkyne cycloaddition reaction cycloaddition reaction.

[0104] In some embodiments, targeting ligands, can be synthesized as activated esters, such as tetrafluorophenyl (TFP) esters, which can be displaced by a reactive amino group (e.g.. NIL-Ce)to attach the targeting ligand to the AR RNAi agents disclosed herein. In some embodiments, targeting ligands are synthesized as azides, which can be conjugated to a propargy l or DBCO group, for example, via Copper (1)- catalyzed or strain-promoted azide-alkyne cycloaddition reaction.

[0105] As shown in Table 5 and 6, above, the example AR RNAi agent sense strand nucleotide sequences are shown to further include, in some embodiments, reactive linking groups at both the 5?terminal end and the 3’ terminal end of the sense strand. For example, certain of the AR RNAi agent sense strand sequences shown in Table 5 or 6 above have an (NH2-C6) linking group at the 5’ end of the nucleotide sequence. Similarly, certain of the AR RNAi agent nucleotide sequences shown in Table 5 or 6 above have a (C6-SS-C6) linking group near the 3’ end of the nucleotide sequence. Such reactive linking groups are positioned to facilitate the linking of targeting ligands, targeting groups, and / or PK / PD modulators to the AR RNAi agents disclosed herein. Linking or conjugation reactions are well known in the art and provide for formation of covalent linkages between two molecules or reactants. Suitable conjugation reactions for use in the scope of the inventions herein include, but are not limited to. amide coupling reaction, Michael addition reaction, hydrazone formation reaction, and click chemistry cycloaddition reaction.

[0106] Additionally, certain of the nucleotide sequences can be synthesized with a dT nucleotide at the 3’ terminal end of the sense strand, followed by (3’ -> 5’) a linker (<?.g., C6-SS-C6). The linker can, in some embodiments, facilitate the linkage to additional components, such as, for example, a lipid or one or more targeting ligands. As described herein, the disulfide bond of C6- SS-C6 is first reduced, removing the dT from the molecule, which can then facilitate the conj ugation of the desired component. The terminal dT nucleotide therefore is not a part of the fully conjugated construct.

[0107] Additionally, certain of the nucleotide sequences can be synthesized with a dT nucleotide at the 3’ terminal end of the sense strand, followed by (3’ -> 5’) a linker (e.g. C6-SS-C6). The linker can, in some embodiments, facilitate the linkage to additional components, such as, for example, an antigen binding protein or one or more targeting ligands. As described herein, the disulfide bond of C6-SS-C6 is first reduced, removing the dT from the molecule, which can then facilitate the conjugation of the desired component. The terminal dT nucleotide therefore is not a part of the fully conjugated construct.

[0108] Additionally, the nucleotide sequences shown in Table 5 and 6 were synthesized with a dT nucleotide at the 3" terminal end of the sense strand, followed by (3’ -> 5’) a linker (e.g.,C6-SS-C6). A suitable and commercially available dT-loaded resin can be used to initiate the synthesis of the oligonucleotide strand. The (C6-SS-C6) linker can. in some embodiments, then be used facilitate the linkage to additional components, such as, for example, a PK / PD modulator or one or more targeting ligands. As described herein, the C6-SS-C6 is first reduced cleaving among other things the dT residue off the molecule, which can then facilitate the conjugation of the desired PK / PD modulator.

[0109] As discussed herein, in some embodiments, one or more targeting ligands and / or PK / PD modulators are linked or conjugated to the RNAi agent. In some embodiments, a targeting ligand (or targeting group) and / or a PK / PD modulator is linked to the 5’ end of the sense strand, the 3’ end of the sense strand, and / or to one or more internal nucleotides. The synthesis of the sense strand and / or the antisense strand can be designed such that reactive groups are readily available to facilitate linkage to additional components, such as a targeting ligand or PK / PD modulator. Table 6.1 depicts the sense strand of the AR RNAi agents disclosed in Table 5 after linking to one or more targeting ligands and / or PK / PD modulators (collectively, shown below, as Z). Pharmacological moieties are linked to the AR RNAi agents using reactions described in Example 1. below. Following conjugation to targeting ligands, the linking groups may have the structure (NH-C6), (NH-C6)s, or (C6-S), the structure of each of which is shown in Table 11.

[0110] In some embodiments, the antisense strand of an AR RNAi agent disclosed herein differs by 0, 1, 2, or 3 nucleotides from any of the antisense strand sequences in Table 3 or Table 10. In some embodiments, the sense strand of an AR RNAi agent disclosed herein differs by 0, 1, 2, or 3 nucleotides from any of the sense strand sequences in Table 4, Table 5. Table 6, or Table 10.

[0111] In some embodiments, an AR RNAi agent antisense strand comprises a nucleotide sequence of any of the sequences in Table 2 or Table 3. In some embodiments, an AR RNAi agent antisense strand comprises the sequence of nucleotides (from 5’ end -> 3‘ end) 1-17, 2-17, 1-18, 2-18, 1-19, 2-19, 1-20, 2-20, 1-21, 2-21, 1-22, 2-22, 1-23, 2-23, 1-24, or 2-24 of any of the sequences in Table 2, Table 3, or Table 10. In certain embodiments, an AR RNAi agent antisense strand comprises or consists of a modified sequence of any one of the modified sequences in Table 3 or Table 10.

[0112] In some embodiments, an AR RNAi agent sense strand comprises the nucleotide sequence of any of the sequences in Table 2 or Table 4. In some embodiments, an AR RNAi agent sense strand comprises the sequence of nucleotides (from 5’ end -> 3’ end) 1-17, 2-17, 3- 17, 4-17, 1-18, 2-18, 3-18, 4-18, 1-19, 2-19, 3-19, 4-19, 1-20, 2-20, 3-20, 4-20, 1-21, 2-21, 3-21 , 4-21, 1-22, 2-22, 3-22, 4-22, 1-23, 2-23, 3-23, 4-23, 1-24, 2-24, 3-24, or 4-24, of any of thesequences in Table 2, Table 4, Table 5. Table 6, or Table 10. In certain embodiments, an AR RNAi agent sense strand comprises or consists of a modified sequence of any one of the modified sequences in Table 3 or Table 10.

[0113] For the RNAi agents disclosed herein, the nucleotide at position 1 of the antisense strand (from 5' end -> 3' end) can be perfectly complementary to an AR gene, or can be non- complementary to an AR gene. In some embodiments, the nucleotide at position 1 of the antisense strand (from 5' end3' end) is a U, A, or dT (or a modified version of U, A or dT). In some embodiments, the nucleotide at position 1 of the antisense strand (from 5’ end 3’ end) forms an A U or U:A base pair with the sense strand.

[0114] In some embodiments, an AR RNAi agent antisense strand comprises the sequence of nucleotides (from 5' end3' end) 2-18 or 2-19 of any of the antisense strand sequences in Table 2, Table 3, or Table 10. In some embodiments, an AR RNAi sense strand comprises the sequence of nucleotides (from 5' end -> 3' end) 1-17 or 1-18 of any of the sense strand sequences in Table 2, Table 4, Table 5, Table 6, or Table 10.

[0115] In some embodiments, an AR RNAi agent includes (i) an antisense strand comprising the sequence of nucleotides (from 5' end3' end) 2-18 or 2-19 of any of the antisense strand sequences in Table 2, Table 3, or Table 10, and (ii) a sense strand comprising the sequence of nucleotides (from 5' end -> 3' end) 1-17 or 1-18 of any of the sense strand sequences in Table 2, Table 4, Table 5, Table 6, or Table 10.

[0116] A sense strand containing a sequence listed in Table 2 or Table 4 can be hybridized to any antisense strand containing a sequence listed in Table 2 or Table 3 provided the two sequences have a region of at least 85% complementarity over a contiguous 16, 17, 18, 19, 20, or 21 nucleotide sequence. In some embodiments, the AR RNAi agent has a sense strand consisting of the modified sequence of any of the modified sequences in Table 4, Table 5, Table 6, or Table 10, and an antisense strand consisting of the modified sequence of any of the modified sequences in Table 3 or Table 10. Certain representative sequence pairings are exemplified by the Duplex ID Nos. shown in Tables 7, 8, and 9.

[0117] In some embodiments, an AR RNAi agent comprises, consists of. or consists essentially of a duplex represented by any one of the Duplex ID Nos. presented herein. In some embodiments, an AR RNAi agent consists of any of the Duplex ID Nos. presented herein. In some embodiments, an AR RNAi agent comprises the sense strand and antisense strand nucleotide sequences of any of the Duplex ID Nos. presented herein. In some embodiments, an AR RNAi agent comprises the sense strand and antisense strand nucleotide sequences of any ofthe Duplex ID Nos. presented herein and a targeting group, linking group, and / or other nonnucleotide group wherein the targeting group, linking group, and / or other non-nucleotide group is covalently linked (i.e., conjugated) to the sense strand or the antisense strand. In some embodiments, an AR RNAi agent includes the sense strand and antisense strand modified nucleotide sequences of any of the Duplex ID Nos. presented herein. In some embodiments, an AR RNAi agent comprises the sense strand and antisense strand modified nucleotide sequences of any of the Duplex ID Nos. presented herein and a targeting group, linking group, and / or other non-nucleotide group, wherein the targering group, linking group, and / or other non-nucleotide group is covalently linked to the sense strand or the antisense strand.

[0118] In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 2, 7, 8, 9, or 10, and comprises a targeting group. In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 2, 7, 8, 9, or 10, and comprises one or more lipid moieties.

[0119] In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 2. 7, 8, 9, or 10, and comprises a lipid moiety7or antigen binding protein. In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 2, 7, 8. 9, or 10, and comprises one or more lipid moieties.

[0120] In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the modified nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 7, 8, 9, and 10.

[0121] In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the modified nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 7, 8, 9, and 10, and comprises a lipid moi ety or antigen binding protein .

[0122] In some embodiments, an AR RNAi agent comprises, consists of. or consists essentially of any of the duplexes of Tables 7, 8, 9, and 10.

[0123] In some embodiments, an AR RNAi agent comprises, consists of, or consists essentially of a duplex represented by any one of the Duplex ID Nos. presented herein. In some embodiments, an AR RNAi agent consists of any of the Duplex ID Nos. presented herein. In some embodiments, an AR RNAi agent comprises the sense strand and antisense strandnucleotide sequences of any of the Duplex ID Nos. presented herein. In some embodiments, an AR RNAi agent comprises the sense strand and antisense strand nucleotide sequences of any of the Duplex ID Nos. presented herein and a targeting group, linking group, antigen binding protein and / or other non-nucleotide group wherein the targeting group, linking group, antigen binding protein and / or other non-nucleotide group is covalently linked (i.e., conjugated) to the sense strand or the antisense strand. In some embodiments, an AR RNAi agent includes the sense strand and antisense strand modified nucleotide sequences of any of the Duplex ID Nos. presented herein. In some embodiments, an AR RNAi agent comprises the sense strand and antisense strand modified nucleotide sequences of any of the Duplex ID Nos. presented herein and a targeting group, linking group, and / or other non-nucleotide group, wherein the targeting group, linking group, antigen binding protein and / or other non-nucleotide group is covalently linked to the sense strand or the antisense strand.

[0124] In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 2. 7, 8, or 9, and comprises an antigen binding protein. In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 2, 7, 8, or 9, and comprises one or more antigen binding proteins.

[0125] In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 2, 7. 8, or 9, and comprises an antigen binding protein. In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 2, 7, 8, or 9, and comprises one or more antigen binding protein.

[0126] In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the modified nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 7, 8, and 9.

[0127] In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Table 2 or Table 6, and further comprises a targeting group. In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the nucleotide sequences of any of the antisense strand / sense strand duplexes of Table 6 (or Table 4, or Table 5), and further comprises an integrin receptor ligand targeting group.

[0128] In some embodiments, an AR RNAi agent comprises an antisense strand and a sense strand having the modified nucleotide sequences of any of the antisense strand / sense strand duplexes of Tables 7, 8, and 9, and comprises an antigen binding protein.

[0129] Table 7. AR RNAi Agent Duplexes with Corresponding Sense and Antisense StrandID Numbers and Sequence ID numbers for the modified and unmodified nucleotide sequences.

[0130] Table 8. AR RNAi Agent Duplexes with Corresponding Sense and Antisense StrandID Numbers and Sequence ID numbers for the modified and unmodified nucleotide sequences.(Shown with Targeting Ligand Conjugates)

[0131] Table 9, Conjugate Duplex ID Numbers Referencing Position Targeted On AndrogenReceptor (AR) Gene

[0132] Table 10. Conjugate ID Numbers With Chemically Modified Antisense and Sense Strands (including Linkers and Conjugates)

[0133] In some embodiments, an AR RNAi agent is prepared or provided as a salt, mixed salt, or a free-acid. In some embodiments, an AR RNAi agent is prepared or provided as a pharmaceutically acceptable salt. In some embodiments, an AR RNAi agent is prepared or provided as a pharmaceutically acceptable sodium or potassium salt The RNAi agents described herein, upon delivery to a cell expressing an AR gene, inhibit or knockdown expression of one or more AR genes in vivo and / or in vitro.Targeting Groups, Linking Groups, Lipid moieties, and Delivery Vehicles

[0134] In some embodiments, an AR RNAi agent contains or is conjugated to one or more non-nucleotide groups including, but not limited to, a targeting group, a linking group, a pharmacokinetic / pharmacodynamic (PK / PD) modulator, a delivery polymer, or a delivery' vehicle. The non-nucleotide group can enhance targeting, delivery', or attachment of the RNAi agent. The non-nucleotide group can be covalently linked to the 3' and / or 5' end of either the sense strand and / or the antisense strand. In some embodiments, an AR RNAi agent contains a non-nucleotide group linked to the 3' and / or 5' end of the sense strand. In some embodiments, a non-nucleotide group is linked to the 5' end of an AR RNAi agent sense strand. A non- nucleotide group can be linked directly7or indirectly7to the RNAi agent via a linker / Iinking group. In some embodiments, a non-nucleotide group is linked to the RNAi agent via a labile, cleavable, or reversible bond or linker.

[0135] In some embodiments, a non-nucleotide group enhances the pharmacokinetic or biodistribution properties of an RNAi agent or conjugate to which it is attached to improve cell- or tissue-specific distribution and cell-specific uptake of the conjugate. In some embodiments, a non-nucleotide group enhances endocytosis of the RNAi agent.

[0136] Targeting groups or targeting moieties enhance the pharmacokinetic or biodistribution properties of a conjugate or RNAi agent to which they are attached to improve cell-specific (including, in some cases, organ specific) distribution and cell-specific (or organ specific) uptake of the conjugate or RNAi agent. A targeting group can be monovalent, divalent, trivalent, tetravalent, or have higher valency for the target to which it is directed. Representative targeting groups include, without limitation, compounds with affinity to cell surface molecule, cell receptor ligands, hapten, antibodies, monoclonal antibodies, antibody fragments, and antibody mimics w'ith affinity7to cell surface molecules. In some embodiments, a targeting group is linked to an RNAi agent using a linker, such as a PEG linker or one, tw o, or three abasic and / or ribitol (abasic ribose) residues, which in some instances can serve as linkers.

[0137] A targeting group, with or without a linker, can be attached to the 5' or 3' end of any of the sense and / or antisense strands disclosed in Tables 2, 3. 4, 5, 6, and 10. A linker, with or without a targeting group, can be attached to the 5' or 3' end of any of the sense and / or antisense strands disclosed in Tables 2, 3, 4, 5. 6, and 10.

[0138] The AR RNAi agents described herein can be synthesized having a reactive group, such as an amino group (also referred to herein as an amine), at the 5'-terminus and / or the 3'- terminus. The reactive group can be used subsequently to attach a targeting moiety using methods typical in the art.

[0139] For example, in some embodiments, the AR RNAi agents disclosed herein are synthesized having an NH2-C6 group at the 5'-terminus of the sense strand of the RNAi agent. The terminal amino group subsequently can be reacted to form a conjugate with, for example, a group that includes a lipid moiety or antigen binding protein. In some embodiments, the AR RNAi agents disclosed herein are synthesized having one or more alky ne groups at the 5'- terminus of the sense strand of the RNAi agent.

[0140] In some embodiments, targeting groups are linked to the AR RN Ai agents without the use of an additional linker. In some embodiments, the targeting group is designed having a linker readily present to facilitate the linkage to an AR RNAi agent. In some embodiments, when two or more RNAi agents are included in a composition, the two or more RNAi agents can be linked to their respective targeting groups using the same linkers. In some embodiments, when tw'O or more RNAi agents are included in a composition, the two or more RNAi agents are linked to their respective targeting groups using different linkers.

[0141] In some embodiments, a linking group is conjugated to the RNAi agent. The linking group facilitates covalent linkage of the agent to a targeting group, pharmacokinetic modulator, delivery polymer, or delivery vehicle. The linking group can be linked to the 3' and / or the 5' end of the RNAi agent sense strand or antisense strand. In some embodiments, the linking group is linked to the RNAi agent sense strand. In some embodiments, the linking group is conjugated to the 5' or 3' end of an RNAi agent sense strand. In some embodiments, a linking group is conjugated to the 5' end of an RNAi agent sense strand. Examples of linking groups, include but are not limited to: C6-SS-C6, 6-SS-6, reactive groups such a primary amines (e.g.. NH2-C6) and alkynes, alkyl groups, abasic residues / nucleotides, ammo acids, tri-alkyne functionalized groups, ribitol, and / or PEG groups. Examples of certain linking groups are provided in Table 11.

[0142] A linker or linking group is a connection between two atoms that links one chemical group (such as an RNAi agent) or segment of interest to another chemical group (such as atargeting group, pharmacokinetic modulator, or delivery' polymer) or segment of interest via one or more covalent bonds. A labile linkage contains a labile bond. A linkage can optionally include a spacer that increases the distance between the two joined atoms. A spacer may further add flexibility and / or length to the linkage. Spacers include, but are not be limited to, alkyl groups, alkenyl groups, alkynyl groups, aryl groups, aralkyl groups, aralkenyl groups, and aralkynyl groups; each of which can contain one or more heteroatoms, heterocycles, amino acids, nucleotides, and saccharides. Spacer groups are well known in the art and the preceding list is not meant to limit the scope of the description. In some embodiments, an AR RNAi agent is conjugated to a polyethylene glycol (PEG) moiety, or to a hydrophobic group having 12 or more carbon atoms, such as a cholesterol or palmitoyl group.

[0143] In some embodiments, an AR RNAi agent is linked to one or more antigen binding proteins. Antigen binding proteins may enhance the bioavailability of the RNAi agent, the delivery of the RNAi agent to a cell of interest, or the facilitation of shuttling the RNAi agent across the blood brain barrier. In some embodiments, the antigen binding protein may be conjugated to a linker at the 3' or 5' end of a sense strand or an antisense strand of an RNAi agent described herein. In some embodiments, an antigen binding protein may be linked at both the 3' or 5' end of either the sense strand or the antisense strand of an RNAi agent described herein.

[0144] In some embodiments, an antigen binding protein may be conjugated to an AR RNAi agent by reacting an AR RNAi agent comprising an amine-comprising linker, for example, (NH2-C6) (see Table 10). In some embodiments, the amine-comprising linker may be located on the 5' end of the sense strand or the antisense strand of an AR RNAi agent. In some embodiments, the amine-comprising linker may be located on the 3' end of the sense strand or the antisense strand of an RNAi agent.

[0145] Any of the AR RNAi agent nucleotide sequences listed in Tables 2, 3, 4, 5, 6, and 9, whether modified or unmodified, can contain 3' and / or 5' targeting group(s), linking group(s), and / or antigen binding fragments. Any of the AR RNAi agent duplexes listed in Tables 7, 8, and 9, whether modified or unmodified, can further compnse a targeting group or linking group, but not limited to, those depicted in Table 10, and the targeting group or linking group can be attached to the 3' or 5' terminus of either the sense strand or the antisense strand of the AR RNAi agent duplex.

[0146] In some embodiments, an AR RNAi agent is linked to one or more lipid moieties. Lipid moieties may enhance the pharmacodynamic or pharmacokinetic properties of the RNAi agent. In some embodiments, the lipid moiety or antigen binding protein may be conjugated to a linkerat the 3' or 5' end of a sense strand or an antisense strand of an RNAi agent described herein. In some embodiments, a lipid moiety or antigen binding protein may be linked at both the 3' or 5' end of either the sense strand or the antisense strand of an RNAi agent described herein.

[0147] In some embodiments, a lipid moiety or antigen binding protein may be conjugated to an AR RNAi agent by reacting an AR RNAi agent comprising an amine-comprising linker, for example, (NH2-C6) (see table 11). In some embodiments, the amine-comprising linker may be located on the 5' end of the sense strand or the antisense strand of an AR RNAi agent. In some embodiments, the amine-comprising linker may be located on the 3' end of the sense strand or the antisense strand of an RNAi agent.

[0148] In some embodiments, an RNAi agent comprising an amine-comprising linker, such as (NH2-C6) or (NH2-C6)s, may be reacted with a lipid comprising an activated ester moiety. Example lipids with activated ester moieties include LP183-p, LP 283-p, LP293-p, LP294-p, LP304-p, LP310-p, LP383-p, LP395-p, and LP396-p as shown in Table 11 below.

[0149] For example, in some embodiments, the AR RNAi agents disclosed herein are synthesized having an NH2-C6 group at the 5'-terminus of the sense strand of the RNAi agent. The terminal amino group subsequently can be reacted to form a conjugate with, for example, a group that includes an antigen binding protein. In some embodiments, the AR RNAi agents disclosed herein are synthesized having one or more alky ne groups at the 5'-terminus of the sense strand of the RNAi agent.

[0150] In some embodiments, an AR RNAi agent may be conjugated to a lipid moiety or antigen binding protein using phosphoramidite synthesis. Synthesizing oligonucleotides using phosphoramidites is well-known in the art. In some embodiments, a lipid moiety or antigen binding protein may be conjugated to the 5' end of the sense strand or the antisense strand of an AR RNAi agent using a phosphoramidite. In some embodiments, a lipid moiety or antigen binding protein may be conjugated to the 3' end of the sense strand or the antisense strand of an AR RNAi agent using a phosphoramidite. In some embodiments, a phosphoramidite selected from (2C8C12)-p, (2C6CI0)-p, LP429 phosphoramidite, HO-C16-p, C16-p, or C22- p, all as shown in Table 11 below', may be used to conjugate a lipid moiety or antigen binding protein to an AR RNAi agent.

[0151] In some embodiments, AR RNAi agents may comprise a lipid moiety or antigen binding protein on an internal nucleotide (i.e., not on the 3' or 5' terminal nucleotides.) In some embodiments, an internal nucleotide may be linked to the 2' position of nbose. In some embodiments AR RNAi agents may comprise aC16, uC16. cC16, or gC16 as shown in Table 11 below'.

[0152] Any of the AR RNAi agent nucleotide sequences listed in Tables 2, 3, 4, 5, 6, and 10, whether modified or unmodified, can contain 3' and / or 5' targeting group(s), linking group(s), and / or lipid moieties. Any of the AR RNAi agent sequences listed in Tables 3, 4, 5, 6, and 10, or are otherwise described herein, which contain a 3' or 5' targeting group, linking group, and / or lipid moiety or antigen binding protein can alternatively contain no 3' or 5' targeting group, linking group, or lipid moiety or antigen binding protein, or can contain a different 3' or 5' targeting group, linking group, or lipid moiety or antigen binding protein including, but not limited to, those depicted in Table 1 1. Any of the AR RNAi agent duplexes listed in Tables 7, 8, 9 and 10, whether modified or unmodified, can further comprise a targeting group or linking group, including, but not limited to, those depicted m Table 11, and the targeting group or linking group can be attached to the 3' or 5' terminus of either the sense strand or the antisense strand of the AR RNAi agent duplex.

[0153] Examples of certain modified nucleotides, capping moieties, lipid moieties, and linking groups are provided in Table 11.Table 11. Structures Representing Various Modified Nucleotides, Capping Moieties. lipid moieties and Linking Groups (wherein indicates the point of connection)

[0154] Alternatively, other linking groups known in the art may be used. In many instances, linking groups can be commercially acquired or alternatively, are incorporated into commercially available nucleotide phosphoramidites. (See, e.g., International Patent Application Publication No. WO 2019 / 161213. which is incorporated herein by reference in its entirety).

[0155] In some embodiments, an AR RNAi agent is delivered without being conjugated to a targeting ligand or pharmacokinetic / pharmacodynamic (PK / PD) modulator (referred to as being “naked” or a “naked RN Ai agent”).

[0156] In some embodiments, an AR RNAi agent is conjugated to a targeting group, a linking group, a PK modulator, and / or another non-nucleotide group to facilitate delivery of the AR RNAi agent to the cell or tissue of choice, for example, to a CNS and / or skeletal muscle cell in vivo. In some embodiments, an AR RNAi agent is conjugated to a lipid moiety or antigen binding protein.

[0157] In some embodiments, an AR RNAi agent is delivered without being conjugated to an antigen binding protein or other targeting group (referred to as being “naked” or a “naked RNAi agent”).

[0158] In some embodiments, an AR RNAi agent is conjugated to a targeting group, a linking group, a PK modulator, and / or another non-nucleotide group to facilitate delivery of the AR RNAi agent to the cell or tissue of choice, for example, to a CNS cell in vivo. In some embodiments, an AR RNAi agent is conjugated to an antigen binding protein.

[0159] In some embodiments, a delivery vehicle may be used to deliver an RNAi agent to a cell or tissue. A delivery vehicle is a compound that improves delivery of the RNAi agent to a cell or tissue. A delivery vehicle can include, or consist of, but is not limited to: a polymer, such as an amphipathic polymer, a membrane active polymer, a peptide, a melittin peptide, a melittin-like peptide (MLP), a lipid, a reversibly modified polymer or peptide, or a reversibly modified membrane active polyamine.

[0160] In some embodiments, the RNAi agents can be combined with lipids, nanoparticles, polymers, liposomes, micelles, DPCs or other delivery systems available in the art for nucleic acid delivery. The RNAi agents can also be chemically conjugated to targeting groups, lipids (including, but not limited to cholesteryl and cholesteryl derivatives), encapsulating m nanoparticles, liposomes, micelles, conjugating to polymers or DPCs (see, for example WO 2000 / 053722, WO 2008 / 022309, WO 201 1 / 104169, and WO 2012 / 083185, WO 2013 / 032829, WO 2013 / 158141, each of which is incorporated herein by reference), by iontophoresis, or by incorporation into other delivery vehicles or systems available in the art such as hydrogels, cyclodextrins, biodegradable nanocapsules, bioadhesive microspheres, or proteinaceous vectors. In some embodiments the RNAi agents can be conjugated to antibodies having affinity for CNS and / or skeletal muscle cells. In some embodiments, the RNAi agents can be linked to targeting ligands that have affinity for CNS and / or skeletal muscle cells or receptors present on CNS and / or skeletal muscle cells.Antigen Binding Proteins

[0001] In one aspect, AR RNAi agents are conjugates to antigen binding proteins. In some embodiments, the antigen binding protein may be selected from the group consisting of: an antibody, an antibody fragment (e.g., an antigen binding fragment, or Fab), scFv, or other functional component or derivative of an antibody encompassing a Fab and / or complementary - determining regions (CDRs) disclosed herein.

[0002] In some embodiments, the antigen binding protein may act as a shuttle to facilitate the crossing of the blood brain barrier (BBB) of the RNAi agent, such that the RNAi agent may be administered subcutaneously and reach CNS tissue. In some embodiments, the antigen binding protein is an anti -Transferrin 1 (TfRl) antibody or Fab.

[0003] In some embodiments, the antigen binding protein is a Fab. In some embodiments, the Fab comprises (i) 6 complementary determining regions (CDRs), (ii) 3 CDRs on the variable light chain (VL), or (iii) 3 CDRs on the variable heavy chain (VH). In some embodiments, the Fab comprises a light constant chain 1 (CL). In some embodiments, the light constant chain 1 (CL) sequence is:RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 717). In some embodiments. the light chain sequence is:DIQLTQSPSSLSASVGDRVTITCRASDKLYSNLAWYQQKPGKAPKLLIYDATLLASGV PSRFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGQGTKVEIKRTVAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTY SLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 718).

[0004] In some embodiments, the Fab comprises a heavy constant chain 1 (CH). In some embodiments, the heavy constant chain 1 (CH) is: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH (SEQ ID NO: 719). In some embodiments the heavy chain sequence is EVQLVESGGGLVQPGGSLRLSCATSGFTFTSYWMHWVRQAPGKGLEWVAEINPTNG RTNYIEKFKSRITLSVDKSKSTVYLQMNSLRAEDTAVYYCARGTRAYHYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH (SEQ ID NO: 720).

[0005] In some embodiments, the antigen binding protein may have a VL CDR1 sequence selected from the group consisting of: RASDGLYSNLA (SEQ ID NO: 721), RASDNLYRNLA (SEQ ID NO: 722), and RASDKLYSNLA (SEQ ID NO: 723).

[0006] In some embodiments, the antigen binding protein may have a VL CDR2 sequence selected from the group consisting of: DATLLAS (SEQ ID NO: 724), DARNLAS (SEQ ID NO: 725), DAFNLAS (SEQ ID NO: 726), DATRLAS (SEQ ID NO: 727), DATKLAS (SEQ ID NO: 728), and DAKNLAS (SEQ ID NO: 729).

[0007] In some embodiments, the antigen binding protein may have a VL CDR 3 sequence of QHFWGTPLT (SEQ ID NO: 730).

[0008] In some embodiments, the antigen binding protein may have a VH CDR1 sequence selected from the group consisting of: GYTFNSYWMH (SEQ ID NO: 731), GYTFKSYWMH (SEQ ID NO: 732), GFTFTSYWMH (SEQ ID NO: 733), GYTFTSYWVH (SEQ ID NO: 734), and GYTFTSYWMH (SEQ ID NO: 735).

[0009] In some embodiments, the antigen binding protein may have a VH CDR2 sequence selected from the group consisting of: EINPTNGRVNYIEKFKS (SEQ ID NO: 736), EINPTNGRFNYIEKFKS (SEQ ID NO: 737), EINPTNGRTNYIEKFKS (SEQ ID NO: 738). and EINPTNGRSNYIEKFKS (SEQ ID NO: 739).

[0010] In some embodiments, the antigen binding protein may have a VH CDR3 sequence of: GTRAYHY (SEQ ID NO: 740).

[0011] In some embodiments, the antigen binding protein comprises a VL sequence of any one of the sequences listed in Table A below. Each of the Fabs described in Table A has a light chain constant sequence of:RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 717)

[0012] Table A. VL chains with CDR mutation combinations.

[0013] In some embodiments, the antigen binding protein comprises a VL sequence of any one of the sequences listed in Table B below. Each of the Fabs described in Table B has a heavy chain constant sequence of:ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH (SEQ ID NO: 719).Table B. VH chains with CDR mutation combinations.

[0014] Tables C-H show the CDR1 and CDR2 variants from VL and VH with the combined beneficial mutations.Table C. VL CDR1 variantTable D. VL CDR2 variantsTable E. VL CDR3 variantTable F. VH CDR1 variantsTable G. VH CDR2 variantsTable H. VH CDR3 variantsTable I. Fab-Fc chain sequences

[0015] In some embodiments. RNAi agents may be conjugated to an antigen binding protein specific to a non-human mammal to carry out studies. RNAi agents may be conjugated to a mouse-specific anti-transferrin antibody such as Fab0016. Fab0016 as used herein refers to an antibody fragment having a light chain sequence of: DIQMTQSPASLSASLEEIVT1TCQASQDIGNWLAWYQQKPGKSPQLLIYGATSLADGV PSRFSGSRSGTQFSLKISRVQVEDIGIYYCLQAYNTPWTFGGGTKLELKRADAAPTVS IFPPSTEQLATGGASVVCLMNNFYPRDISVKWKIDGTERRDGVLDSVTDQDSKDSTY SMSSTLSLTKADYESHNLYTCEVVHKTSSSPVVKSFNRNEC (SEQ ID NO: 761), and a heavy chain sequence of:EVQLVESGGGLVQPGNSLTLSCVASGFTFSNYGMHWIRQAPKKGLEWIAMIYYDSS KMNYADTVKGRFT1SRDNSKNTLYLEMNSLRSEDTAMYYCAVPTSHYVVDVWGQGVSVTVSSAETTAPSVYPLAPGTALKSNSMVTLGCLVKGYFPEPVTVTWNSGALSSGVHTFPAVLQSGLYTLTSSVTVPSSTWSSQAVTCNVAHPASSTKVDKKIVPREC (SEQ IDNO: 762)Pharmaceutical Compositions and Formulations

[0161] The AR RNAi agents disclosed herein can be prepared as pharmaceutical compositions or formulations (also referred to herein as "‘medicaments’’). In some embodiments, pharmaceutical compositions include at least one AR RNAi agent. These pharmaceutical compositions are particularly useful in the inhibition of the expression of AR mRNA in a target cell, a group of cells, a tissue, or an organism. The pharmaceutical compositions can be used to treat a subject having a disease, disorder, or condition that would benefit from reduction in the level of the target mRNA, or inhibition in expression of the target gene. The pharmaceutical compositions can be used to treat a subject at risk of developing a disease or disorder that would benefit from reduction of the level of the target mRNA or an inhibition in expression the target gene. In one embodiment, the method includes administering an AR RNAi agent linked to a targeting ligand or an antigen binding protein as described herein, to a subject to be treated. In some embodiments, one or more pharmaceutically acceptable excipients (including vehicles, carriers, diluents, and / or delivery' polymers) are added to the pharmaceutical compositions that include an AR RNAi agent, thereby forming a pharmaceutical formulation or medicament suitable for in vivo delivery' to a subject, including a human

[0162] The pharmaceutical compositions that include an AR RNAi agent and methods disclosed herein decrease the level of the target mRNA in a cell, group of cells, group of cells, tissue, organ, or subject, including by administering to the subject a therapeutically effective amount of a herein described AR RNAi agent, thereby inhibiting the expression of AR mRNA in the subject. In some embodiments, the subject has been previously identified or diagnosed as having a disease or disorder that can be mediated at least in part by a reduction in AR expression. In some embodiments, the subject has been previously diagnosed with having one or more neurodegenerative diseases such as SBMA. In some embodiments the neurodegenerative disease is SBMA.

[0163] In some embodiments the subject has been previously diagnosed with having neurodegenerative disease.

[0164] Embodiments of the present disclosure include pharmaceutical compositions for delivering an AR RNAi agent to a CNS and / or skeletal muscle cell in vivo. Such pharmaceuticalcompositions can include, for example, an AR RNAi agent conjugated to a lipid moiety or antigen binding moiety.

[0165] In some embodiments, the described pharmaceutical compositions including an AR RNAi agent are used for treating or managing clinical presentations in a subject that would benefit from the inhibition of expression of AR. In some embodiments, a therapeutically or prophylactically effective amount of one or more of pharmaceutical compositions is administered to a subject in need of such treatment. In some embodiments, administration of any of the disclosed AR RNAi agents can be used to decrease the number, severity, and / or frequency of symptoms of a disease in a subject.

[0166] In some embodiments, the described AR RNAi agents are optionally combined with one or more additional (i.e., second, third, etc.) therapeutics. A second therapeutic can be another AR RNAi agent (e.g., an AR RNAi agent that targets a different sequence within an AR gene). In some embodiments, a second therapeutic can be an RNAi agent that targets the AR gene. An additional therapeutic can also be a small molecule drug, antibody, antibodyfragment, and / or aptamer. The AR RNAi agents, with or without the one or more additional therapeutics, can be combined with one or more excipients to form pharmaceutical compositions.

[0167] The described pharmaceutical compositions that include an AR RNAi agent can be used to treat at least one symptom in a subject having a disease or disorder that would benefit from reduction or inhibition in expression of AR mRNA. In some embodiments, the subject is administered a therapeutically effective amount of one or more pharmaceutical compositions that include an AR RNAi agent thereby treating the symptom. In other embodiments, the subject is administered a prophylactically effective amount of one or more AR RNAi agents, thereby preventing or inhibiting the at least one symptom.

[0168] In some embodiments, one or more of the described AR RNAi agents are administered to a mammal in a pharmaceutically acceptable carrier or diluent. In some embodiments, the mammal is a human.

[0169] The route of administration is the path by which an AR RNAi agent is brought into contact with the body. In general, methods of administering drugs, oligonucleotides, and nucleic acids, for treatment of a mammal are well known in the art and can be applied to administration of the compositions described herein. The AR RNAi agents disclosed herein can be administered via any suitable route in a preparation appropriately tailored to the particular route. Thus, in some embodiments, the herein described pharmaceutical compositions are administered via inhalation, intranasal administration, intratrachealadministration, or oropharyngeal aspiration administration. In some embodiments, the pharmaceutical compositions can be administered by injection, for example, intravenously, intramuscularly, intracutaneously, subcutaneously, intracerebroventricularly. intraarticularly, intraocularly, or intraperitoneally, or topically.

[0170] The pharmaceutical compositions including an AR RNAi agent described herein can be delivered to a cell, group of cells, tissue, or subject using oligonucleotide delivery technologies known in the art. In general, any suitable method recognized in the art for delivering a nucleic acid molecule (in vitro or in vivo) can be adapted for use with the compositions described herein. For example, delivery can be by local administration, (e.g., direct injection, implantation, or topical administering), systemic administration, or subcutaneous, intravenous, intraperitoneal, or parenteral routes, including intracranial (e.g., intraventricular, intraparenchymal and intrathecal), mtracerebroventricular, intramuscular, transdermal, airway (aerosol), nasal, oral, rectal, or topical (including buccal and sublingual) administration. In some embodiments, the compositions are administered via inhalation, intranasal administration, oropharyngeal aspiration administration, or intratracheal administration. For example, in some embodiments, it is desired that the AR RNAi agents described herein inhibit the expression of an AR gene in the CNS or skeletal muscle tissue.

[0171] In some embodiments, the pharmaceutical compositions described herein comprise one or more pharmaceutically acceptable excipients. The pharmaceutical compositions described herein are formulated for administration to a subject.

[0172] As used herein, a pharmaceutical composition or medicament includes a pharmacologically effective amount of at least one of the described therapeutic compounds and one or more pharmaceutically acceptable excipients. Pharmaceutically acceptable excipients (excipients) are substances other than the Active Pharmaceutical Ingredient (API, therapeutic product, e.g., AR RNAi agent) that are intentionally included in the drug delivery' system. Excipients do not exert or are not intended to exert a therapeutic effect at the intended dosage. Excipients can act to a) aid in processing of the drug delivery' system during manufacture, b) protect, support or enhance stability, bioavailability or patient acceptability of the API, c) assist in product identification, and / or d) enhance any' other attribute of the overall safety, effectiveness, of delivery of the API during storage or use. A pharmaceutically acceptable excipient may or may not be an inert substance.

[0173] Excipients include, but are not limited to: absorption enhancers, anti-adherents, antifoaming agents, anti-oxidants, binders, buffering agents, carriers, coating agents, colors, delivery enhancers, delivery polymers, detergents, dextran, dextrose, diluents, disintegrants,emulsifiers, extenders, fillers, flavors, glidants, humectants, lubricants, oils, polymers, preservatives, saline, salts, solvents, sugars, surfactants, suspending agents, sustained release matrices, sweeteners, thickening agents, tonicity agents, vehicles, water-repelling agents, and wetting agents.

[0174] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water-soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor® ELTM (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). It should be stable under the conditions of manufacture and storage and should be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof The proper fluidity' can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, sugars, poly alcohols such as mannitol, sorbitol, and sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0175] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filter sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic di spersion medium and the required other ingredi ents from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation include vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0176] Formulations suitable for intra-articular administration can be in the form of a sterile aqueous preparation of the drug that can be in microcrystalline form, for example, in the form of an aqueous microcrystalline suspension. Liposomal formulations or biodegradable polymer systems can also be used to present the drug for both intra-articular and ophthalmic administration.

[0177] The active compounds can be prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, includingimplants and microencapsulated deliver}' systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled m the art. Liposomal suspensions can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Patent No 4,522,811.

[0178] The AR RNAi agents can be formulated in compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form refers to physically discrete units suited as unitary' dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the disclosure are dictated by and directly dependent on the unique characteristics of the active compound and the therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.

[0179] A pharmaceutical composition can contain other additional components commonly found in pharmaceutical compositions. Such additional components include, but are not limited to: anti-pruritics, astringents, local anesthetics, or anti-inflammatory' agents (e.g., antihistamine, diphenhydramine, etc.). It is also envisioned that cells, tissues, or isolated organs that express or comprise the herein defined RNAi agents may be used as “pharmaceutical compositions.” As used herein, “pharmacologically effective amount,” “therapeutically effective amount,” or simply “effective amount” refers to that amount of an RNAi agent to produce a pharmacological, therapeutic, or preventive result.

[0180] In some embodiments, the methods disclosed herein further comprise the step of administering a second therapeutic or treatment in addition to administering an RNAi agent disclosed herein. In some embodiments, the second therapeutic is another AR RNAi agent (e.g., an AR RNAi agent that targets a different sequence within the AR target). In other embodiments, the second therapeutic can be a small molecule drug, an antibody , an antibodyfragment, and / or an aptamer.

[0181] In some embodiments, described herein are compositions that include a combination or cocktail of at least two AR RNAi agents having different sequences. In some embodiments, the two or more AR RNAi agents are each separately and independently linked to lipids.

[0182] Described herein are compositions for delivery' of AR RNAi agents to central nervous system cells. Furthermore, compositions for delivery of AR RNAi agents to cells, including neurons, astrocytes, microglia and endothelial cells, in vivo, are generally described herein

[0183] Generally, an effective amount of an AR RNAi agent disclosed herein will be in the range of from about 0.0001 to about 20 mg / kg of body weight / deposited dose, e.g., from about 0.001 to about 5 mg / kg of body weight / deposited dose. In some embodiments, an effective amount of an AR RNAi agent will be in the range of from about 0.01 mg / kg to about 3.0 mg / kg of body weight per deposited dose. In some embodiments, an effective amount of an AR RNAi agent will be in the range of from about 0.03 mg / kg to about 2.0 mg / kg of body weight per deposited dose. In some embodiments, an effective amount of an AR RNAi agent will be in the range of from about 0.01 to about 1.0 mg / kg of deposited dose per body weight. In some embodiments, an effective amount of an AR RNAi agent will be in the range of from about 0.50 to about 1.0 mg / kg of deposited dose per body weight. The amount administered will also likely depend on such variables as the overall health status of the patient, the relative biological efficacy of the compound delivered, the formulation of the drug, the presence and types of excipients in the formulation, and the route of administration. Also, it is to be understood that the initial dosage administered can be increased beyond the above upper level to rapidly achieve the desired blood-level or tissue level, or the initial dosage can be smaller than the optimum. In some embodiments, a dose is administered daily. In some embodiments, a dose is administered weekly. In further embodiments, a dose is administered bi-weekly, tri-weekly, once monthly, or once quarterly (i.e., once even7three months).

[0184] For treatment of disease or for formation of a medicament or composition for treatment of a disease, the pharmaceutical compositions described herein including an AR RNAi agent can be combined with an excipient or with a second therapeutic agent or treatment including, but not limited to: a second or other RNAi agent, a small molecule drug, an antibody, an antibody fragment, peptide, and / or an aptamer.

[0185] The described AR RNAi agents, when added to pharmaceutically acceptable excipients or adjuvants, can be packaged into kits, containers, packs, or dispensers.Methods of T reatment and Inhibition of AR Expression

[0186] The AR RNAi agents disclosed herein can be used to treat a subject (e.g., a human or other mammal) having a disease or disorder that would benefit from administration of the RNAi agent. In some embodiments, the RNAi agents disclosed herein can be used to treat a subject (e.g., a human) that would benefit from a reduction and / or inhibition in expression of AR mRNA and / or a reduction in AR receptor levels.

[0187] In some embodiments, the RNAi agents disclosed herein can be used to treat a subject (e.g., a human) having a disease or disorder for which the subject would benefit from reductionin androgen receptor, including but not limited to, SBMA. Treatment of a subject can include therapeutic and / or prophylactic treatment. The subject is administered a therapeutically effective amount of any one or more AR RNAi agents described herein. The subject can be a human, patient, or human patient. The subject may be an adult, adolescent, child, or infant. Administration of a pharmaceutical composition described herein can be to a human being or animal.

[0188] Mutant AR activity is known to promote neurodegenerative disorders. In some embodiments, the described AR RNAi agents are used to treat at least one symptom mediated at least in part by a reduction in mutant AR levels, in a subject. The subject is administered a therapeutically effective amount of any one or more of the described AR RNAi agents. In some embodiments, the subject is administered a prophylactically effective amount of any one or more of the described RNAi agents, thereby treating the subject by preventing or inhibiting the at least one symptom.

[0189] In certain embodiments, the present disclosure provides methods for treatment of diseases, disorders, conditions, or pathological states mediated at least in part by AR gene expression, in a patient in need thereof, wherein the methods include administering to the patient any of the AR RNAi agents described herein.

[0190] In some embodiments, the AR RNAi agents are used to treat or manage a clinical presentation or pathological stale in a subject, wherein the clinical presentation or pathological state is mediated at least in part by a reduction in AR expression The subject is administered a therapeutically effective amount of one or more of the AR RNAi agents or AR RNAi agentcontaining compositions described herein. In some embodiments, the method comprises administering a composition comprising an AR RNAi agent described herein to a subject to be treated.

[0191] In a further aspect, the disclosure features methods of treatment (including prophylactic or preventative treatment) of diseases or symptoms that may be addressed by a reduction in AR receptor levels, the methods comprising administering to a subject in need thereof an AR RNAi agent that includes an antisense strand comprising the sequence of any of the sequences in Table 2, Table 3, or Table 10. Also described herein are compositions for use in such methods.

[0192] The described AR RNAi agents and / or compositions that include AR RNAi agents can be used in methods for therapeutic treatment of disease or conditions caused by enhanced or elevated AR receptor activity levels. Such methods include administration of an AR RNAi agent as described herein to a subject, e.g., a human or animal subject.

[0193] In another aspect, the disclosure provides methods for the treatment (including prophylactic treatment) of a pathological state (such as a condition or disease) mediated at least in part by AR expression, wherein the methods include administering to a subject a therapeutically effective amount of an RNAi agent that includes an antisense strand comprising the sequence of any of the sequences in Table 2, Table 3, or Table 10.

[0194] In some embodiments, methods for inhibiting expression of an AR gene are disclosed herein, wherein the methods include administering to a cell an RNAi agent that includes an antisense strand comprising the sequence of any of the sequences in Table 2, Table 3, or Table 10.

[0195] In some embodiments, methods for the treatment (including prophylactic treatment) of a pathological state mediated at least in part by AR expression are disclosed herein, wherein the methods include administering to a subject a therapeutically effective amount of an RNAi agent that includes a sense strand comprising the sequence of any of the sequences in Table 2, Table 4, Table 5, Table 6, or Table 10.

[0196] In some embodiments, methods for inhibiting expression of an AR gene are disclosed herein, wherein the methods comprise administering to a cell an RNAi agent that includes a sense strand comprising the sequence of any of the sequences in Table 2, Table 4, Table 5, Table 6, or Table 10.

[0197] In some embodiments, methods for the treatment (including prophylactic treatment) of a pathological state mediated at least in part by AR expression are disclosed herein, wherein the methods include administering to a subject a therapeutically effective amount of an RNAi agent that includes a sense strand comprising the sequence of any of the sequences in Table 4, Table 5, Table 6, or Table 10, and an antisense strand comprising the sequence of any of the sequences in Table 3 or Table 10.

[0198] In some embodiments, methods for inhibiting expression of an AR gene are disclosed herein, wherein the methods include administering to a cell an RNAi agent that includes a sense strand comprising the sequence of any of the sequences in Table 4, Table 5, Table 6, or Table 10, and an antisense strand comprising the sequence of any of the sequences in Table 3 or Table 10.

[0199] In some embodiments, methods of inhibiting expression of an AR gene are disclosed herein, wherein the methods include administering to a subject an AR RNAi agent that includes a sense strand consisting of the nucleobase sequence of any of the sequences in Table 4, Table 5, Table 6, or Table 10, and the antisense strand consisting of the nucleobase sequence of any of the sequences in Table 3 or Table 10. In other embodiments, disclosed herein are methodsof inhibiting expression of an AR gene, wherein the methods include administering to a subject an AR RNAi agent that includes a sense strand consisting of the modified sequence of any of the modified sequences in Table 4, Table 5, Table 6, or Table 10, and the antisense strand consisting of the modified sequence of any of the modified sequences in Table 3 or Table 10.

[0200] In some embodiments, methods for inhibiting expression of an AR gene in a cell are disclosed herein, wherein the methods include administering one or more AR RNAi agents comprising a duplex structure of one of the duplexes set forth in Tables 7, 8, 9. and 10.

[0201] In some embodiments, the gene expression level and / or mRNA level of an AR gene in certain CNS cells and / or skeletal muscle cells of subject to whom a described AR RNAi agent is administered is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%. 55%. 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%. 97%. 98%, 99%, or greater than 99%, relative to the subject prior to being administered the AR RNAi agent or to a subject not receiving the AR RNAi agent. In some embodiments, the AR receptor or AR protein levels in certain CNS and / or skeletal muscle cells of a subject to whom a described AR RNAi agent is administered is reduced by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%. 55%. 60%. 65%, 70%, 75%. 80%, 85%. 90%. 95%. 96%. 97%. 98%, 99%, or greater than 99%, relative to the subject prior to being administered the AR RNAi agent or to a subject not receiving the AR RNAi agent. Hie gene expression level, protein level, and / or mRNA level in the subject may be reduced in a cell, group of cells, and / or tissue of the subject. In some embodiments, the AR mRNA levels in certain CNS and / or skeletal muscle cells subject to whom a described AR RNAi agent has been administered is reduced by at least about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%. 85%, 90%, 95%. or 98% relative to the subject prior to being administered the AR RNAi agent or to a subject not receiving the AR RNAi agent.

[0202] A reduction in gene expression, mRNA, and protein levels can be assessed by any methods known in the art. Reduction or decrease in AR receptor activity level and / or AR protein levels are collectively referred to herein as a decrease in, reduction of, or inhibition of AR expression. The Examples set forth herein illustrate known methods for assessing inhibition of AR expression and AR gene expression.Cells, Tissues, Organs, and Non-Human Organisms

[0203] Cells, tissues, organs, and non-human organisms that include at least one of the AR RNAi agents described herein are contemplated. The cell, tissue, organ, or non-humanorganism is made by delivering the RNAi agent to the cell, tissue, organ, or non-human organism.Additional Illustrative Embodiments

[0204] Provided here are certain additional illustrative embodiments of the disclosed technology. These embodiments are illustrative only and do not limit the scope of the present disclosure or of the claims attached hereto.1. An RNAi agent for inhibiting expression of an Androgen Receptor (AR) gene, comprising: an antisense strand comprising at least 17 contiguous nucleotides differing by 0 or 1 nucleotides from any one of the sequences provided in Table 2 or Table 3; and a sense strand comprising a nucleotide sequence that is at least partially complementary to the antisense strand.2. The RNAi agent of embodiment 1. wherein the antisense strand comprises nucleotides 2-18 of any one of the sequences provided in Table 2 or Table 3.3. The RNAi agent of embodiment 1 or embodiment 2, wherein the sense strand comprises a nucleotide sequence of at least 17 contiguous nucleotides differing by 0 or 1 nucleotides from any one of the sequences provided in Table 2 or Table 4, and wherein the sense strand has a region of at least 85% complementarity over the 17 contiguous nucleotides to the antisense strand.4. The RNAi agent of any one of embodiments 1-3, wherein at least one nucleotide of the AR RNAi agent is a modified nucleotide or includes a modified intemucleoside linkage.5. The RNAi agent of any one of embodiments 1-4, wherein all or substantially all of the nucleotides are modified nucleotides.6. The RNAi agent of any one of embodiments 4-5, wherein the modified nucleotide is selected from the group consisting of: 2'-O-methyl nucleotide, 2'-fluoro nucleotide, 2'-deoxy nucleotide, 2’,3'-seco nucleotide mimic, locked nucleotide, 2'-F-arabino nucleotide, 2'-methoxy ethyl nucleotide, abasic nucleotide, ribitol, inverted nucleotide, inverted 2'-O-methyl nucleotide, inverted 2'-deoxy nucleotide, 2'-amino-modified nucleotide, 2'-alkyl-modified nucleotide, morpholino nucleotide, vinyl phosphonate-containing nucleotide, cyclopropyl phosphonate-containing nucleotide, and 3'-O-methyl nucleotide.7. The RNAi agent of embodiment 5. wherein all or substantially all of the nucleotides are modified with 2'-O-methyl nucleotides, 2'-fluoro nucleotides, or combinations thereof.8. The RNAi agent of any one of embodiments 1-7, wherein the antisense strand comprises the nucleotide sequence of any one of the modified sequences provided in Table 3.9. The RNAi agent of any one of embodiments 1-8, wherein the sense strand comprises the nucleotide sequence of any one of the modified sequences provided in Table 4.10. The RNAi agent of embodiment 1. wherein the antisense strand comprises the nucleotide sequence of any one of the modified sequences provided in Table 3 and the sense strand comprises the nucleotide sequence of any one of the modified sequences provided in Table 4.11. The RNAi agent of any one of embodiments 1-10. wherein the sense strand is between 18 and 30 nucleotides in length, and the antisense strand is between 18 and 30 nucleotides in length.12. The RNAi agent of embodiment 11, wherein the sense strand and the antisense strand are each between 18 and 27 nucleotides in length.13. The RNAi agent of embodiment 12, wherein the sense strand and the antisense strand are each between 18 and 24 nucleotides in length.14. The RNAi agent of embodiment 13, wherein the sense strand and the antisense strand are each 21 nucleotides in length.15. The RNAi agent of embodiment 14, wherein the RNAi agent has two blunt ends.16. The RNAi agent of any one of embodiments 1-15. wherein the sense strand comprises one or two terminal caps.17. The RNAi agent of any one of embodiments 1-16, wherein the sense strand comprises one or two inverted abasic residues.18. The RNAi agent of embodiment 1. wherein the RNAi agent comprises of a sense strand and an antisense strand that form a duplex having the structure of any one of the duplexes in Table 7, Table 8, Table 9, or Table 10.19. The RNAi agent of embodiment 18, wherein all or substantially all of the nucleotides are modified nucleotides.20. The RNAi agent of embodiment 19, wherein all or substantially all of the nucleotides are modified nucleotides.21. The RNAi agent of any one of embodiments 19-20, wherein the sense strand further includes inverted abasic residues at the 3’ terminal end of the nucleotide sequence, at the 5’ end of the nucleotide sequence, or at both.22. The RNAi agent of any one of embodiments 1 -21 , wherein the RNAi agent is linked to a lipid moiety or linked to an antigen binding protein.23. The RNAi agent of any one of embodiments 1-22. wherein the RNAi agent is linked to a lipid moiety.24. The RNAi agent of embodiment 23, wherein the lipid moiety is selected from the group consisting of:wherein ? indicates the point of connection to the RNAi agent.25. The RNAi agent of embodiment 23 or embodiment 24, wherein the lipid moiety is conjugated to the sense strand.26. The RNAi agent of embodiment 25, wherein the lipid moiety is conjugated to the 5’ terminal end of the sense strand.27. The RNAi agent of any of embodiments 1-22, wherein the RNAi agent is linked to an antigen binding protein.28. The RNAi agent of any one of embodiments 1-27. wherein the antigen binding protein is an antibody fragment (Fab) that specifically binds to one or more epitopes on a transferrin receptor (TfRl).29. The RNAi agent of embodiment 28, wherein the Fab comprises (i) 6 complementary determining regions (CDRs), (ii) 3 CDRs on the variable light chain (VL), and / or (iii) 3 CDRs on the variable heavy chain (VH).30. The RNAi agent of embodiment 29, wherein the variable light chain has a VL CDR1 sequence selected from the group consisting of: RASDGLYSNLA (SEQ ID NO: 721), RASDNLYRNLA (SEQ ID NO: 722), and RASDKLYSNLA (SEQ ID NO: 723); a VL CDR2 sequence selected from the group consisting of: DATLLAS (SEQ ID NO: 724), DARNLAS (SEQ ID NO: 725), DAFNLAS (SEQ ID NO: 726), DATRLAS (SEQ ID NO: 727). DATKLAS (SEQ ID NO: 728), and DAKNLAS (SEQ ID NO: 729); and / or a VL CDR 3 sequence of QHFWGTPLT (SEQ ID NO: 730).31. The RNAi agent of embodiment 29 or 30, wherein the variable light chain comprises the sequence:DIQLTQSPSSLSASVGDRVTITCRASDKLYSNLAWYQQKPGKAPKLLIYDATLLASGV PSRFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGQGTKVEIK (SEQ ID NO: 747).32. The RNAi agent of any one of embodiments 29-31, wherein the variable heavy chain has a VH CDR1 sequence selected from the group consisting of GYTFNSYWMH (SEQ ID NO: 731), GYTFKSYWMH (SEQ ID NO: 732), GFTFTSYWMH (SEQ ID NO: 733), GYTFTSYWVH (SEQ ID NO: 734), and GYTFTSYWMH (SEQ ID NO: 735), a VH CDR2 sequence selected from the group consisting of: EINPTNGRVNYIEKFKS (SEQ ID NO: 736). EINPTNGRFNYIEKFKS (SEQ ID NO: 737), EINPTNGRTNYIEKFKS (SEQ ID NO: 738), and EINPTNGRSNYIEKFKS (SEQ ID NO: 739); and / or a VH CDR3 sequence of: GTRAYHY (SEQ ID NO: 740).33. The RNAi agent of any one of embodiments 29-32, wherein the variable heavy chain comprises the sequence:EVQLVESGGGLVQPGGSLRLSCATSGFTFTSYWMHWVRQAPGKGLEWVAEINPTNG RTNYIEKFKSRITLSVDKSKSTVYLQMNSLRAEDTAVYYCARGTRAYHYWGQGTLV TVSS (SEQ ID NO: 755).34. The RNAi agent of any one of embodiments 28-33, wherein the Fab further comprises a light constant chain 1 (CL).35. The RNAi agent of embodiment 34 wherein the light constant chain 1 (CL) sequence is:RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 717).36. The RNAi agent of any one of embodiments 28-35, wherein the Fab further comprises a heavy constant chain 1 (CH).37. The RNAi agent of embodiment 36 wherein the heavy constant chain 1 (CH) sequence is:ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH (SEQ ID NO: 719).38. The RNAi agent of any one of embodiments 28-37, wherein the antibody fragment (Fab) binds TfRl with an affinity of at least 1 nM KD.39. A conjugate comprising the RNAi agent of any one of embodiments 1-21 conjugated to an antibody fragment (Fab) that specifically binds to one or more epitopes on a transferrin receptor (TfRl).40. The conjugate of embodiment 39, wherein the Fab comprises (i) 6 complementary determining regions (CDRs), (ii) 3 CDRs on the variable light chain (VL), or (iii) 3 CDRs on the variable heavy chain (VH).41. The conjugate of embodiment 40, wherein the variable light chain has a VL CDR1 sequence selected from the group consisting of: RASDGLYSNLA (SEQ ID NO: 721), RASDNLYRNLA (SEQ ID NO: 722). and RASDKLYSNLA (SEQ ID NO: 723); a VL CDR2 sequence selected from the group consisting of: DATLLAS (SEQ ID NO: 724), DARNLAS (SEQ ID NO: 725), DAFNLAS (SEQ ID NO: 726), DATRLAS (SEQ ID NO: 727), DATKLAS (SEQ ID NO: 728), and DAKNLAS (SEQ ID NO: 729); and / or a VL CDR 3 sequence of QHFWGTPLT (SEQ ID NO: 730).42. The conjugate of embodiment 40 or 41, wherein the variable light chain comprises the sequence:DIQLTQSPSSLSASVGDRVTITCRASDKLYSNLAWYQQKPGKAPKLLIYDATLLASGV PSRFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGQGTKVEIK (SEQ ID NO: 747).43. The conjugate of any one of embodiments 40-42, wherein the variable heavy chain has a VH CDR1 sequence selected from the group consisting of: GYTFNSYWMH (SEQ ID NO: 731), GYTFKSYWMH (SEQ ID NO: 732). GFTFTSYWMH (SEQ ID NO: 733), GYTFTSYWVH (SEQ ID NO: 734), and GYTFTSYWMH (SEQ ID NO: 735). a VH CDR2 sequence selected from the group consisting of: EINPTNGRVNYIEKFKS (SEQ ID NO: 736), EINPTNGRFNYIEKFKS (SEQ ID NO: 737), EINPTNGRTNYIEKFKS (SEQ ID NO: 738), and EINPTNGRSNYIEKFKS (SEQ ID NO: 739); and / or a VH CDR3 sequence of: GTRAYHY (SEQ ID NO: 740).44. The conjugate of any one of embodiments 40-43, wherein the variable heavy chain comprises the sequence:EVQLVESGGGLVQPGGSLRLSCATSGFTFTSYWMHWVRQAPGKGLEWVAEINPTNG RTNYIEKFKSRITLSVDKSKSTVYLQMNSLRAEDTAVYYCARGTRAYHYWGQGTLV TVSS (SEQ ID NO: 755)45. The conjugate of any one of embodiments 39-44 wherein the Fab further comprises a light constant chain 1 (CL).46. The conjugate of embodiment 45 wherein the light constant chain 1 (CL) sequence is: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 717).47. The conjugate of any one of embodiments 39-46, wherein the Fab further comprises a heavy constant chain 1 (CH).48. The conjugate of embodiment 47 wherein the heavy constant chain 1 (CH) sequence is: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH (SEQ ID NO: 719).49. The conjugate of any one of embodiments 39-48, wherein the antibody fragment (Fab) binds TfRl with an affinity of at least 1 nM KD.50. The conjugate of embodiment any one of embodiments 39-49, wherein the RNAi agent is conjugated to the Fab using a covalent or non-covalent bond, ionic bond, hydrogen bond, hydrophobic interaction, peptide, polymer, or a nucleic acid binding protein.51. The conjugate of any one of embodiments 39-50. wherein the RNAi agent is conjugated to the Fab through a linker comprising a structure selected from the group consisting of:wherein5A represents a point of attachment to the Fab, and *R represents a point of attachment to the RNAi agent portion of the conjugate.52. The RNAi agent of any one of embodiments 1 -21 , wherein the RNAi agent is linked to a targeting ligand.53. The RNAi agent of embodiment 52, wherein the targeting ligand is linked to the sense strand.54. The RNAi agent of embodiment 53, wherein the targeting ligand is linked to the 5’ terminal end of the sense strand.55. The RNAi agent of any of embodiments 52-54, wherein the targeting ligand has affinity for a skeletal muscle cell and / or a cell receptor expressed on a skeletal muscle cell.56. The RNAi agent of any of embodiments 52-55, wherein the targeting ligand has affinity for an integrin alpha-v-beta 6 (av06) receptor.57. The RNAi agent of any of embodiments 52-56, wherein the targeting ligand is:peptide 1), or a pharmaceutically acceptable salt thereof, wherein ? indicates the point of connection to the RNAi agent.58. The RNAi agent of any one of embodiments 1-21 or 52-57, wherein the RNAi agent is further linked to a pharmacokinetic / pharmacodynamic (PK / PD) modulator.59. The RNAi agent of embodiment 58, wherein the PK / PD modulator is linked to the sense strand.60. The RNAi agent of embodiment 59. wherein the PK / PD modulator is linked to the 3’ terminal end of the sense strand.61. The RNAi agent of any one of embodiments 58-60, wherein the PK / PD modulator is selected from the group consisting of:wherein Rz comprises the RNAi agent.62. A composition comprising the RNAi agent of any one of embodiments 1-38 or 52-61, or the conjugate of any one of embodiments 39-51, wherein the composition further comprises a pharmaceutically acceptable excipient.63. The composition of embodiment 62, further comprising a second RNAi agent capable of inhibiting the expression of Androgen Receptor gene expression.64. The composition of any one of embodiments 62-63, further comprising one or more additional therapeutics.65. The composition of any of embodiments 62-64, wherein the RNAi agent is a sodium salt.66. The composition of any of embodiments 62-65, wherein the pharmaceutically acceptable excipient is water for injection.67. The composition of any of embodiments 62-65, wherein the pharmaceutically acceptable excipient is a buffered saline solution.68. A method for inhibiting expression of an AR gene in a cell, the method comprising introducing into a cell an effective amount of the RNAi agent of any one of embodiments 1- 38 or 52-61, the conjugate of any one of embodiments 39-51, or the composition of any one of embodiments 62-67.69. The method of embodiment 68, wherein the cell is within a subject.70. The method of embodiment 69, wherein the subject is a human subject.71. The method of any one of embodiments 68-70, wherein following the administration of the RNAi agent the Androgen Receptor (AR) gene expression is inhibited by at least about 30%.72. A method of treating one or more symptoms or diseases associated with enhanced or elevated membrane AR activity' levels, the method comprising administering to a human subject in need thereof a therapeutically effective amount of the composition of any one of embodiments 62-67.73. The method of embodiment 72, wherein the disease is a neurodegenerative disease.74. The method of embodiment 73, wherein the neurodegenerative disease is spinal and bulbar muscular atrophy (SBMA).75. The method of any one of embodiments 68-74, wherein the RNAi agent is administered at a deposited dose of about 0.01 mg / kg to about 5.0 mg / kg of body weight of the subj ect.76. The method of any one of embodiments 68-75, wherein the RNAi agent is administered at a deposited dose of about 0.03 mg / kg to about 2.0 mg / kg of body weight of the subj ect.77. The method of any of embodiments 68-76, wherein the RNAi agent is administered in two or more doses.78. Use of the RNAi agent of any one of embodiments 1-38 or 52-61. or the conjugate of any one of embodiments 39-51, for the treatment of a disease, disorder, or symptom that is mediated at least in part by mutant AR activity and / or AR gene expression.79. Use of the composition according to any one of embodiments 62-67, for the treatment of a disease, disorder, or symptom that is mediated at least in part by Androgen Receptor (AR) activity and / or Androgen Receptor (AR) gene expression.80. Use of the composition according to any one of embodiments 62-67, for the manufacture of a medicament for treatment of a disease, disorder, or symptom that is mediated at least in part by Androgen Receptor (AR) and / or Androgen Receptor (AR) gene expression.81. The use of any one of embodiments 78-80, wherein the disease is a neurodegenerative disease.82. A method of making an RNAi agent of any one of embodiments 1-38 or 52-61, comprising annealing a sense strand and an antisense strand to form a double-stranded ribonucleic acid molecule.83. The method of embodiment 82, wherein the sense strand comprises a lipid moiety.84. The method of embodiment 82, wherein the sense strand comprises an antigen binding moiety.85. The method of embodiment 82, wherein the sense strand comprises a targeting ligand.86. The method of embodiment 85, wherein the sense strand further comprises a PK / PD modulator.87. The method of embodiment 83, comprising conjugating a lipid moiety to the sense strand.88. The method of embodiment 84, comprising conjugating an antigen binding moiety to the sense strand.89. The method of embodiment 85, comprising conjugating a targeting ligand to the sense strand.90. The method of embodiment 86, comprising conjugating a PK / PD modulator to the sense strand.EXAMPLESExample 1. Synthesis of AR RNAi Agents.

[0205] AR RNAi agent duplexes disclosed herein were synthesized in accordance with the following:

[0206] A. Synthesis. The sense and antisense strands of the AR RNAi agents were synthesized according to phosphoramidite technology on solid phase used in oligonucleotide synthesis. Depending on the scale, a MerMade96E® (Bioautomation), a MerMadeI2® (Bioautomation), or an OP Pilot 100 (GE Healthcare) was used. Syntheses were performed on a solid support made of controlled pore glass (CPG, 500 A or 600A, obtained from Prime Synthesis, Aston, PA, USA). All RNA and 2'-modified RNA phosphoramidites were purchased from Thermo Fisher Scientific (Milwaukee. WI, USA). Specifically, the 2'-O-methyl phosphoramidites that were used included the following: (5'-O-dimethoxytntyl-N6-(benzoyl)-2'- O-methyl-adenosine-3'-O-(2-cyanoethyl-N,N-diisopropylamino) phosphoramidite, 5'-O- dimethoxy-trityl-N4-(acetyl)-2'-O-methyl-cytidine-3'-O-(2-cyanoethyl-N,N-diisopropyl- amino) phosphoramidite, (5'-0-dimethoxytrityl-N2-(isobutyryl)-2'-0-methyl-guanosine-3'-0- (2-cyanoethyl-N,N-diisopropylamino) phosphoramidite, and 5'-O-dimethoxytrityl-2'-O- methyl-uridine-3'-O-(2-cyanoethyl-N,N-diisopropylamino) phosphoramidite. The 2'-deoxy-2'- fluoro-phosphoramidites carried the same protecting groups as the 2'-O-methyl RNA amidites. 5'-dimelhoxytrityl-2'-O-methyl-inosine-3'-O-(2-cyanoethyl-N,N-diisopropylamino) phosphoramidites were purchased from Glen Research (Virginia). The inverted abasic (3'-O- dimethoxytrityl-2'-deoxyribose-5'-O-(2-cyanoethyl-N,N-diisopropylamino) phosphoramidites were purchased from ChemGenes (Wilmington, MA, USA). Hie following UNA phosphoramidites were used: 5'-(4,4'-Dimethoxytrityl)-N6-(benzoyl)-2’,3'-seco-adenosine, 2'- benzoyl-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, 5'-(4,4'-Dimethoxytrityl)-N- acetyl-2',3 '-seco-cytosine, 2'-benzoyl-3'-[(2-cyanoethyl)-(N,N-diiso-propyl)]- phosphoramidite, 5'-(4,4l-Dimethoxytrityl)-N-isobutyiy-l-2',3'-seco-guanosine, 2'-benzoyl-3'- [(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, and 5'-(4,4'-Dimethoxy-trityl)-2',3'- seco-undine, 2’-benzoyl-3'-[(2-cyanoethyl)-(N,N- diiso-propyl)] -phosphoramidite. TFA aminolink phosphoramidites were also commercially purchased (ThermoFisher). Linker L6 was purchased as propargyl-PEG5-NHS from BroadPharm (catalog # BP-20907) and coupled to the NH2-C6 group from an aminolink phosphoramidite to form -L6-C6-, using standard coupling conditions. The linker Alk-cyHex was similarly commercially purchased from Lumiprobe (alkyne phosphoramidite, 5?-terminal) as a propargyl-containing compound phosphoramidite compound to form the linker -Alk-cyHex-. In each case, phosphorothioatelinkages were introduced as specified using the conditions set forth herein. The cyclopropyl phosphonate phosphorami dites were synthesized in accordance with International Patent Application Publication No. WO 2017 / 214112 (see also Altenhofer et. al., Chem. Communications (Royal Soc. Chem.), 57(55):6808-6811 (July 2021)).

[0207] Tri-alkyne-containing phosphoramidites were dissolved in anhydrous dichloromethane or anhydrous acetonitrile (50 mM), while all other amidites were dissolved in anhydrous acetonitrile (50 mM) and molecular sieves (3A) were added. 5-Benzylthio-lH-tetrazole (BTT. 250 mM in acetonitrile) or 5-Ethylthio-lH-tetrazole (ETT, 250 mM in acetonitrile) w7as used as activator solution. Coupling times were 10 minutes (RNA), 90 seconds (2' O-Me), and 60 seconds (2' F). In order to introduce phosphorothioate linkages, a 100 mM solution of 3-phenyl l,2,4-dithiazoline-5-one (POS, obtained from PolyOrg, Inc., Leominster, MA, USA) in anhydrous acetonitrile was employed.

[0208] Alternatively, tri-alkyne moieties were introduced post-synthetically (see section E, below). For this route, the sense strand was functionalized with a 5' and / or 3' terminal nucleotide containing a primary amine. TFA aminolink phosphoramidite was dissolved in anhydrous acetonitrile (50 mM) and molecular sieves (3 A) were added. 5-Benzylthio-lH- tetrazole (BTT, 250 mM in acetonitrile) or 5-Ethylthio-lH-tetrazole (ETT, 250 mM in acetonitrile) was used as activator solution. Coupling times were 10 minutes (RNA), 90 seconds (2' O-Me), and 60 seconds (2' F). In order to introduce phosphorothioate linkages, a 100 mM solution of 3-phenyl l,2,4-dithiazoline-5-one (POS, obtained from PolyOrg. Inc., Leominster. MA, USA) in anhydrous acetonitrile was employed.

[0209] B. Cleavage and deprotection of support bound oligomer. After finalization of the solid phase synthesis, the dried solid support was treated with a 1: 1 volume solution of 40 wt. % methylamine in w7ater and 28% to 31% ammonium hydroxide solution (Aldrich) for 1.5 hours at 30°C. The solution was evaporated and the solid residue was reconstituted in water (see below7).

[0210] C. Purification. Crude oligomers were purified by anionic exchange HPLC using aTSKgel SuperQ-5PW 13pm column and Shimadzu LC-8 system. Buffer A was 20 mM Tris, 5 mM EDTA, pH 9.0 and contained 20% Acetonitrile and buffer B was the same as buffer A with the addition of 1.5 M sodium chloride. UV traces at 260 nin were recorded. Appropriate fractions were pooled then run on size exclusion HPLC using a GE Healthcare XK 16 / 40 column packed with Sephadex G-25 fine with a running buffer of lOOmM ammonium bicarbonate, pH 6.7 and 20% Acetonitrile or filtered water. Alternatively, pooled fractions weredesalted and exchanged into an appropriate buffer or solvent system via tangential flow7filtration.

[0211] D. Annealing. Complementary strands were mixed by combining equimolar RNA solutions (sense and antisense) in l x PBS (Phosphate-Buffered Saline, l x, Coming, Cellgro) to form the RNAi agents. Some RNAi agents were lyophilized and stored at -15 to -~25°C. Duplex concentration was determined by measuring the solution absorbance on a UV-Vis spectrometer in l x PBS. The solution absorbance at 260 nm was then multiplied by a conversion factor (0.050 mg / (mL-cm)) and the dilution factor to determine the duplex concentration.

[0212] E. Synthesis of Lipids

[0213] If lipids described herein are not included in Example IE, it is to be assumed that the compounds are commercially available. For example, LP395p and LP396p were purchased.

[0214] Synthesis of LP183 phosphoramidite

[0215] To a solution of compound 2 (2.00 g) in DCM was added TEA (2.27 mL) followed by compound 1 (4.931 g) dropwise at room temperature. Then the mixture was stirred at room temperature for 2h. The mixture was then filtered. The white solid was dried overnight. Product is as white solid, yield. 4.267g, 74%. LC-MS: calculated [M+H] 356.35, found 356.63.

[0216] To a mixture of compound 1 (2.54 g) in 120 mL DCM was added compound 3 (0.61 g) followed by compound 2 (5.37 g) drop wise at room temperature. Then the mixture was stirred at room temperature overnight. 5 mL TEA was added followed by Celite. After removing solvent in vacuo, the residue was loaded on a 40g column by dry method. Hexanes (2% TEA)to 50% EtOAc (2% TEA) in Hexanes (2% TEA) as gradient was used to purify the product. Product is a white waxy solid, yield 3.462 g, 87%. LC-MS: calculated [M+H] 556.46, found 556.64.

[0218] To a solution of compound 1 (49 mg), NEts (0.068 mL), and COMU (76.8 mg) in DMF was added compound 2 (29.8 mg) under ambient conditions. The reaction was stirred until full conversion was observed by LC-MS. Conversion was not able to be clearly observed by LC- MS, and instead, reaction was allowed to stir for 30 min. until bright yellow^ color (before the addition of compound 2) transitioned to a honey orange color and all material was observed to be mainly dissolved. The reaction mixture was washed with water, extracted with DCM, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by CombiFlash® via DCM liquid-load onto a 12-g column with a gradient hexanes to 100% EtOAc in which product eluted at 31% B. The product was concentrated under vacuum to provide a white solid residue and confirmed by 1H NMR in CDCh.

[0219] Synthesis of LP293-p U

[0220] To a solution of compound 1 (73 mg), NEts (0. 112 mL). and COMU (126 mg) in DMF was added compound 2 (48.9 mg) under ambient conditions. The reaction was stirred until full conversion was observed by LC-MS. Conversion was not able to be clearly observ ed by LC- MS, and instead, reaction was allowed to stir for 30 min. until bright yellow color (before theaddition of compound 2) transitioned to a honey orange color and all material was observed to be mainly dissolved. The reaction mixture was then washed with water, extracted with DCM, dried over Na-2SO4, filtered, and concentrated under vacuum. The residue was purified by CombiFlash® via DCM liquid-load onto a 12-g column with a gradient hexanes to 100% EtOAc in which product eluted at 30% B. The product w as concentrated under vacuum to provide a white solid residue and confirmed by 1H NMR in CDCls.

[0221] Synthesis of LP294-p U

[0222] To a solution of compound 1 (40.3 mg), NEt? (0.062 mb), and COMU (69.7 mg) in DCM was added compound 2 (27.0 mg) under ambient conditions. The reaction was stirred until full conversion was observed by LC-MS. Conversion was not able to be clearly observed by LC-MS. and instead, reaction was allowed to stir for 30 min. until bright yellow' color (before the addition of compound 2) transitioned to a honey orange color and all material w'as observed to be mainly dissolved. The reaction mixture w as directly concentrated for isolation. The residue was purified by CombiFlash® via DCM liquid-load onto a 12-g column with a gradient hexanes to 100% EtOAc in which product eluted at 32% B. The product was concentrated under vacuum to provide a white solid residue and confirmed by 1H NMR in CDCh.

[0223] Synthesis of LP-304p

[0224] 1 (200 mg, 0.377 mmol, 1.0 eqv.) was hydrolyzed with LiOH (151 mg, 3.77 mmol, 10.0 eqv.) in MeOH / TFH / H2O(l : 1 : 1 , 90 mL). After removing all organic solvent, the aqueous phase was acidified to pH=3 with IN HC1. The reaction mixture extracted with ethyl acetate (100 mL x3). The organic phases were combined, dried with Na2SO4 and concentrated to get crude acid.

[0225] To a solution of above crude acid and tetrafluorophenol 4 (68.9 mg, 0.415 mmol, 1.10 eqv.) in 30 mL DCM was added COML (194 mg, 0.453 mmol, 1.20 eqv.) and then TEA (0. 158 mL, 1.13 mmol, 3.0 eqv.) under ambient conditions. The reaction was stirred until full conversion was observed by LC-MS. The reaction mixture was washed with IN HC1, then brine. Dry with Na-zSOr and concentrated. The reaction mixture was purified by CombiFlash® using silica gel as the stationary phase with a gradient of EA to Hex 0-100%. 170 mg product was obtained (85% yield).

[0226] Synthesis of LP-310p

[0227] To the solution of 1 in DCM was added DIPEA (0.057 mL), COMU (0.077 g) and 2 (0.0300 g) at room temperature. After stirring at room temperature for 2h, the reaction was quenched with 0. IN HC1. The organic layer was washed with brine. After removing the solvent, the residue was loaded on a 4g column. Hexanes to 50% Hexanes in EtOAc as gradient was used to purify. Product was a white solid, 46mg, 44%. LC-MS: calculated [M+H] 422.36, found 422.61.

[0228] The solution of 1 (0.046 g) in 4N HCl / Dioxane (2 mL) was stirred at room temperature overnight. After removing the solvent in vacuo, the residue was placed under high vacuum for 3h. Then the residue was dissolved in DCM at room temperature, then COMU (0.0700 g), DIPEA (0.038 mL) and 2 (0.036 g) were added at room temperature. After stirring at room temperature for 2h, the solvent was removed in vacuo. The residue was loaded on a 4g column. Hexanes to 50% Hexanes in EtOAc as gradient was used to purify. Product was a white solid, 21mg, 38%. LC-MS: calculated [M+H] 514.29, found 514.61.

[0229] Synthesis of LP383-p

[0230] To the solution of compound 1 (0.050 g) in 5 mL DCM was added compound 2 (0.023 g) and EDC (0.039 g) at room temperature. The mixture was stirred at room temperature for Ih. After removing the solvent in vacuo, the residue was loaded on a 4g column by dry method. Hexanes to 50%EtOAc in Hexanes was used to pwify the product. Pdt is a white solid, yield, 29mg. LC-MS: calculated [M+H+H20] 388.27, found 388.03.

[0231] Synthesis of LP409 phosphoramidite

[0232] Compounds 1 (1.40 g) and 2 (0.613 g) were dissolved in 100 mL THF, then TEA (2.01 mL) was added. The reaction was stirred at 60°C until full conversion was confirmed via LC- MS (2-3 hows). The reaction was cooled down to room temperatwe. Product obtained as whilte precipitate, which was filtered and washed with Acetone (20mL). Compound structwe was verified using ’H and,3P NMR.

[0233] Compounds 1 (1.9 g), 2 (0.846 g) and 3 (2.98 g) were dissolved in 100 mL DCM then heated to 40°C. The reaction was stirred until the solution became clear. The reaction was cooled down to room temperature and stirred overnight. After removing all DCM, the product was dry loaded onto a 24g column. Product was obtained as a white solid using 0-50% (EA / Hex, 1% TEA added) as mobile phase.

[0234] Synthesis of (2C8C12) phosphoramidite

[0235] 2-ocytyl-l -decanol (1.00 grams, 3.35 mmol) and diisopropylamonium tetrazolide (0.2868 grams, 1.68 mmol) were placed in a flask and the flask was purged with nitrogen. DCM (50 mL) was added to the mixture and 2-Cyanoethyl A,A,MA"- tetraisopropylphosphorodiamidite (2.66 mL, 8.37 mmol) w'as added dropwise. Upon completion of the reaction. 3 mL of triethylamine was added to the reaction and the reaction was concentrated directly onto celite for purification. The crude product w as purified buy silica gel chromatography (0: 100 EtOAc:hexanes + 2% triethylamine to 100:0 EtOAc:Hexanes + 2% tri ethylamine) The product eluted w'ith 100% Hexanes. Fractions containing product were concentrated to 1.268 g (76% yield) of a clear liquid.

[0236] Synthesis of (2C6C10) phosphoramidite

[0237] 2-hexy 1-1 -decanol (1.00 g, 4.13 mmol) and diisopropylamonium tetrazolide (0.353 g, 2.06 mmol) were placed in a flask and the flask was purged with nitrogen. DCM (50 mL) was added to the mixture and 2-Cyanoethyl N,N,N',N'-tetraisopropylphosphorodiamidite (3.27 mL, 10.3 mmol) was added dropwise. . Upon completion of the reaction, 3 mL of triethyl amine was added to the reaction and the reaction was concentrated directly onto celite for purification. The crude product was purified buy silica gel chromatography (0:100 EtOAc: hexanes + 2% triethylamine to 100:0 EtOAc:Hexanes + 2% triethylamine) The product eluted with 100% Hexanes. Fractions containing product were concentrated to 1.32 g (72% yield) of a clear liquid.

[0238] Synthesis of HO-C16 phosphorami di te

[0239] 1,16-hexadecanediol, N,N-diisopropylethylamine (0.100 g) was dissolved in 2 mL THF. 4,4“ -Dimethoxy trityl chloride (2.2 g, 6.6 mmol) was added slowly as a solid. After 2 h, the reaction was concentrated by rotary evaporation, and the product was purified by column chromatography (25% ethyl acetate / 75% hexane).

[0240] DMT-O-Cis-OH (0.200 g), Bis(diisopropylamino)(2-cyanoethoxy)phosphine (0.227 mL) and BisDiisopropylammonium tetrazolide (0.0611 g) were dissolved in anhydrous DCM at room temperature. The reaction was capped and stirred overnight. Conversion was determined via LC-MS (0.25M NH4 HCOcHpO buffer system). Celite® was added to the reaction mixture and it was concentrated under vacuum until a white powder remained. The mixture was loaded dry onto a silica column (12 gram) using a EtOAc / Hexanes (1% Triethylamine) solvent system to prevent hydrolysis from the silica gel.[1]The product was characterized by31PNMR, ^NMR, and LC-MS.

[0241] Synthesis of C16 phosporamidite

[0242] Cetyl alcohol (1.10 g), Bis(diisopropylamino)(2-cyanoethoxy)phosphine (2.88 mL) and BisDiisopropylammonium tetrazolide (0.778 g) were dissolved in a solution of DCM at room temperature. The reaction was capped and stirred overnight. Conversion was determined via LC-MS (0.25M NEU HCO3 H2O buffer system). C elite® was added to the reaction mixture and it was concentrated under vacuum until a white powder remained. The mixture was loaded dry onto a silica column (12 gram) using a s EtOAc / Hexanes (1% Triethylamine) solvent system to prevent hydrolysis from the silica gel. The desired product was not retained on the column and came out shortly after being loaded. The isolated product was then charaterized by LC-MS, ‘HNMR and31PNMR. Final yield: 856.5 mg (93.8%).

[0243] Synthesis of C22 phosporamidite

[0244] Docosanol (1.10 g), Bis(diisopropylamino)(2-cyanoethoxy)phosphine (2.1 mL) and BisDiisopropylammonium tetrazolide (0.577 g) were dissolved in a solution of DCM at room temperature. The reaction was capped and stirred overnight. Conversion was determined via LC-MS (0.25M NH4 HCOvFLO buffer system). Celite® was added to the reaction mixtureand it was concentrated under vacuum until a white powder remained. The mixture was loaded dry onto a silica column (12 gram) pretreated with 3 mL of triethylamine using a EtOAc / Hexanes (1% Triethylamine) solvent system to prevent hydrolysis from the silica gel. The isolated product was then charaterized by LC-MS, ’HNMR and3lPNMR. Final yield: 2.1085 g (118.8%).

[0245] Synthesis of phosphorami dite for aC16 internal nucleotide

[0246] Compound 2: Sodium hydride (60% dispersion in mineral oil, 4.17g, 104 mmol) was added in 2 portions ( 15 min apart) into a cold solution of adenosine 1 ( 12.5 g, 46.77 mmol) in anhydrous DMF (250 mL). The cooling bath was removed, the reaction mixture was stirred for1.5h at RT and 1 -bromohexadecane (18 g, 59 mmol) was added. Following 16 h of stirring at RT ethanol (5 mL) was added, stirred for 15 min, DMF was removed on a rotavapor, and toluene w?as evaporated twice to get nd of residual DMF. The product was isolated on CombiFlash following solid load with 40g of silica gel using 220g SiCh column. Eluent: DCM (A) - 20% MeOH in DCM (B), B = 0 - 20%, 15 min, then 20% for 5 min. Product was dried in vacuo ON. Yield 2.726 g. Calculated: MW 491.68. Found: MS (ES, positive): 492.46 [M+H]+.

[0247] Compound 3: A solution of compound 2 (2.7 g, 5.49 mmol) in anhydrous pyridine (40 mL) was cooled on an ice bath and TMS-C1 (2.25 mL, 17.73 mmol) was added. The reaction mixture was stirred for 30 min on ice bath, benzoyl chloride (1.5 mL, 12.9 mmol) was added, the cooling bath w as removed in 30 min, stirring was continued overnight. In 16 h the reaction mixture was cooled on ice bath, water (7.5 mL) was added, stirred for additional 30 min. Concentrated NITiOH (7.5 mL) was added and all volatiles were removed on a rotavapor. To get rid of side bis-acylated adduct, the crude material was dissolved in MeOH (125 mL) and treated with NH4OH (13 mL) for 25 min. The solvent was removed on a rotavapor and toluene was evaporated once. CombiFlash® purification was performed using solid load with silica gel (18 g) on 80 g column using eluent: DCM - 20% MeOH in DCM, 0 - 20%, 50 min. Yield 2.66g. Calculated: MW 595.79 Found: MS (ES, positive): 596.53 [M+H]+.

[0248] Compound 4: Compound 3 (1.55g, 2.60 mmol) was dried by successive evaporations of toluene and anhydrous pyridine using dry' rotavapor. It was dissolved in anhydrous pyridine (10 mL), and DMAP (12 mg, 0. 1 mmol) was added followed by dimethoxy trity l chloride (965 mg, 2.9 mmol). The reaction was stirred for 16 h at RT. All volatiles were removed on a rotavapor, the residual pyridine was removed by evaporation of toluene. The residue was partitioned between DCM and aqueous NaHCCh. The organic phase was separated, the aqueous was extracted with DCM, combined organic phases yvere dried (Na2SO4) and concentrated. The product was isolated on CombiFlash® using 40 g column, eluent: hexane (A) - ethyl acetate (B) + 1 % of Et3N, B= 20-60%, 40 min. Yield 1.845 g. Calculated: MW 898.16. Found: MS (ES, positive): 899.65 [M+H]+.

[0249] Compound 5: Compound 4 (1.845 g. 2.052 mmol) yvas dried by 2 evaporations of toluene. It was dissolved in anhydrous DCM (30 mL), diisopropylammonium tetrazolide (176 mg, 1.03 mmol) and dry molecular sieves (100 mg) yvere added and stirred for 30 min. 2- Cyanoethyl N,N,N’,N'-tetraisopropylphosphorodiamidite (804 mg, 2.67 mmol) was added and stirring w as continued for 16 h. The reaction mixture was diluted 3 times w ith anhydrous DCM, filtered, and stirred with 150 mL of cold NaFICOs solution for 5 min. Organic phase w as separated, aqueous yvas extracted with DCM, combined organic phases were washed withNaHCCE, and dried (Na?SO4). The product was isolated on CombiFlash® using 40 g column, eluent: hexane (A) - ethyl acetate (B) + 1% of E13N. B= 15-60%. 30 min. Yield 1.386 g. Calculated: MW 1098.38. Found: MS (ES, positive): 1099.17 [M+HJ+.

[0250] Synthesis of phosphorami dite for gC 16 internal nucleotide4

[0251] Compound 2: A solution of N2-lsobutyrylguanosine 1 (5g, 14.15 mmol) in anhydrous DMF (120 mL) was added into a cold flask with sodium hydride (60% dispersion in mineral oil, 1.3g, 32.55 mmol). The cooling bath was removed, the reaction mixture was stirred for 3h at RT and 1 -bromohexadecane (5.61 g, 18.4 mmol) was added. The reaction mixture was stirred at 50°C for 72 h and EtOH (2 mL) was added. DMF was removed on a rotavapor, and toluene was evaporated twice to get rid of residual DMF. The product was isolated on CombiFlash® following solid load with 30g of silica gel using 120g column. System for separation: A= DCM : EtOAc (1 : 1); B = DCM:EtOAc:MeOH (9:9:2). B=0 - 100%, 50 min.Kept fraction B = 32-70%. Yield 2.45 g.Calculated: MW 577.77. Found: MS (ES, positive): 579.39 [M+H]+.

[0252] Compound 3: Compound 2 (2. 129 g, 3.69 mmol) was dried by 2 evaporations of toluene followed by one evaporation of anhydrous pyridine. It was dissolved in anhydrous pyridine (40 mL), dimethoxytrityl chloride (1.375 g, 4.06 mmol) and DMAP (18 mg, 0.148 mmol) were added and stirred overnight. All volatiles were removed on a rotavapor followed by 2 evaporations of toluene. The residue was taken m DCM (150 mL) and stirred with NaHCCh (50 mL) for 3 min. The organic phase was separated, the aqueous phase was extracted with DCM, combined organic phases were w ashed with NaHCCh, and dried (Na2SO4). The product was isolated on CombiFlash® using 40 g column, eluent: hexane (A) - ethyl acetate (B) + 1% of Et3N. B= 20-90%. 40 min. Kept fraction B= 58-70%. Yield 1.606 g. Calculated: MW 880.14. Found: MS (ES. positive): 881.34 [M+HJ+.

[0253] Compound 4: Compound 3 (1.4 g, 1.59 mmol) was dried by 2 evaporations of toluene. It was dissolved in anhydrous DCM (30 mL), diisopropylammonium tetrazolide (136 mg, 0.8 mmol) and dry molecular sieves (100 mg) were added and stirred for 30 min. 2-Cyanoethyl N,N.N',N'-tetraisopropylphosphorodiamidite (767 mg, 2.54 mmol) was added and stirring was continued for 16 h. EtjN (0.3 mL) w?as added followed by silica gel (6g). The mixture was concentrated in vacuo and solid loaded on CombiFlash®. Product was isolated using 40 g column, eluent: hexane(A) - ethyl acetale(B) + 1% of ElsN, B= 20-90%, 35 min. Yield 836 mg. Calculated: MW 1080.36. Found: MS (ES, positive): 1081.38 [M+H]+.

[0254] Synthesis of phosphorami dite for uC16 internal nucleotide

[0255] Compound 2: Dry 2.2‘-Cyclouridine 1 (18 g, 79.6 mmol) and DMAP (486 mg. 3.98 mmol) were suspended in anhydrous pyridine (120 mL) and treated with tert- butyl(chloro)diphenylsilane (25.6 g, 93.14 mmol). The reaction mixture was stirred at RT for 48 h. Pyridine was removed on a rotavapor, The residual pyridine was co-evaporated with toluene. The residue was dissolved in ethyl acetate (200 mL), washed with 10% H3PO4 (75 mL), 5% NaCl, and brine, then dried with Na2.SO4. Compound 2 was isolated following CombiFlash® purification using two 120 g SKh columns, eluent DCM (A) - 20% MeOH (B) in DCM, B= 0-60%. Yield 14 g. Calculated: MW 464.59. Found: MS (ES, positive): 465.71 [M+H]+.

[0256] Compound 3: Hexadecanol (49.6 g, 204.4 mmol) was dried in vacuo overnight. It was dissolved in dry diglyme (38 mL) with heating and cooled to RT. AlMes (2M solution in heptane, 31.25 mL, 62.5 mmol) was slowly added under flow of N2 and the reaction mixture was heated at 110°C until methane evolution ceased (30 min). It was cooled to RT, and dry uridine derivative 2 (13.2 g, 28.39 mmol) was added as a solid, dry diglyme was added (30 mL). The reaction was heated at 140°C for 18 h. It was partitioned between 10% H3PO4 (300 mL) and EtOAc (200 mL). The aqueous layer was extracted with EtOAc; combined organicphases were washed twice with 5% NaCl, brine, and dried with Na2.SO4. Following filtration and concentration the crude solid was dried by evaporation of toluene and kept in vacuo overnight. Crude product 3 (115.8 g) was directly used in the following step.

[0257] Compound 4: Crude compound 3 was dissolved in THF (130 mL), triethylamine trishy drofluoride (18 mL, 114 mmol) was added, and the reaction mixture was stirred for 3 days. The product was partitioned between EtOAc (300 mL) and 5% aqueous NaCl (250 mL). Organic phase was separated, the aqueous phase was extracted with EtOAc, combined organic phases were washed with brine, dried (Na?.SO4), and concentrated to dryness. Product 4 was isolated following CombiFlash® purification on two 220 g SiO2 columns applying solid load with 60 g of silica gel. Eluent: DCM (A), 10% MeOH in DCM (B), B= 0-60% in 60 min. Product was dried by 2 evaporations of toluene. Yield 5.63 g. Calculated: MW 468.64. Found: MS (ES, positive): 469.52 [M+H]+; 491.52 [M+Na]+.

[0258] Compound 5: Compound 4 (5.63 g, 12 mmol), 4,4’-dimethoxytrytyl chloride (5.2 g, 15.36 mmol), and DMAP (293 mg, 2.4 mmol) were dissolved in anhydrous pyridine (40 mL), EtsN (2.17 mL, 15.36 mmol) was added and stirred for 16 h. The reaction was quenched with MeOH (0.6 mL), stirred for 15 min, pyridine was removed in vacuo, the residue was partitioned between EtOAc (230 mL) and 5% aqueous NaCl (230 mL). The organic phase was separated, aqueous phase was extracted with EtOAc, combined organic phases were washed with brine and dried overNa2SO4. CombiFlash® purification using 120g SiO2 column, eluent: Hexane(A) -EtOAc (B), B= 10-50% 60 min. Yield 8.271 g. Calculated: MW 771.01. Found: MS (ES, positive): 772. 35 [M+H]+.

[0259] Compound 6: Compound 5 (2.33 g, 3.024 mmol) was dried by 2 evaporations of anhydrous ACN and placed in high vacuo for 2h. The dry compound 5 was dissolved in anhydrous DCM (40 mL) and stirred with diisopropylammonium tetrazolide (704 mg. 4.11 mmol) and molecular sieves for 20 min. 2-Cyanoethyl N,N,N',N'- tetraisopropylphosphorodiamidite (1.404 g, 4.66 mmol) was added, the reaction mixture was stirred for 16 h at RT, diluted to 100 ml with anhydrous DCM, the solid was filtered, and shaken with cold NaHCOs (100 mL) for 5 min. The organic layer wfas separated, washed with NaHCCh, dried (Na2.SO4) and product was purified on CombiFlash® using 40 g SiCh column. Eluent: hexane (A) - EtOAc (B) with 1% EtsN, B= 15-50?4 Yield 2g. Calculated: MW 971.23. Found: MS (ES, positive): 972.17 [M+H]+.

[0260] Synthesis of phos phorami dite for cC16 internal nucleotide

[0261] Compound 2: 5'-O-DMT-protected Uracil -derivative 1, described in preparation of cC16 (11.687g, 15.15 mmol) was dissolved in anhydrous pyridine (100 mL) and cooled on ice bath. TMS-C1 (7.8 mL, 61 mmol) was added, the cooling bath was removed, reaction was stirred for 30 min at RT and cooled on an ice bath again.

[0262] In a separate flask 1.2,4-triazole (35 g. 50.7 mmol) was suspended in anhydrous CAN (200 mL), cooled on ice bath, POCh (11.2 mL, 120 mmol) was slowly added over 10 min, and stirred for 5 min at 0°C. EtsN (84 mL, 595 mmol) was added slowly over 30 min and stirred at 0°C for 30 min.

[0263] The cooled content with silyl ated Uracil derivative from the fist flask was quickly added in one portion to the mixture with 1, 2,4-triazole, stirred for lOmin and cooling bath was removed. The stirring was continued for 5h at RT. The reaction mixture was concentrated to 1 / 3 of its volume on a rotavapor, diluted with EtOAc (600 mL), and washed with 5% NaCl (2 x 400 mL). An aqueous phase was back-extracted with EtOAc (200 mL). Combined EtOAclayers were washed with brine and dried (NajSOr). The EtOAc solution was filtered and concentrated and dried in vacuo to obtain crude derivative 2 (17.46 g).

[0264] Compound 3: Cmde compound 4 was dissolved in dry dioxane (220 mL) in thick wall IL RB flask, concentrated ammonium hydroxide solution (50 mL) was added, the flask was sealed with rubber septa, and the reaction mixture was stirred for 40 h at RT. All volatiles were removed on a rotavapor. Toluene was evaporated twice to dry the residue. Product was isolated on a CombiFlash following solid load with 35g of silica gel using 220g SiCh column. Eluent: DCM (A) - 10%MeOH in DCM (B), B= 0 - 45%, 60 min. Yield 7.81. Calculated: MW 770.02. Found: MS (ES, positive): 770.31 [M+H]+, 1541.24 [2M+H]+.

[0265] Compound 4: Compound 3 (6.94 g, 9 mmol) was dried by evaporation of anhydrous DMF and was dissolved in anhydrous DMF (60 mL). Acetic anhydride (1.75 mL, 18 mmol) was added and stirred at RT for 16 h. NaHCCh solution (250 mL) was added and product w as extracted with DCM (2 x 200 mL). It w?as washed with brine (50 mL), dried (Na2SO4), concentrated and dried by 2 successive evaporations of toluene. CombiFlash® purification w?as performed using 120 g SiO?. column, eluent: DCM (A), 5%MeOH in DCM (B), B= 0-40% 60 min. Yield 6.55 g. Calculated: MW 812.06. Found: MS (ES, positive): 813 36 [M+H]+.

[0266] Compound 5: Compound 4 (6.55 g, 8.07 mmol) was dried by 2 evaporations of toluene. It was dissolved in anhydrous DCM (166 mL) and stirred with diisopropylammonium tetrazolide (2.188 g, 12.78 mmol) and molecular sieves (500 mg) for 20 min. 2-Cyanoethyl N,N,N',N'-tetraisopropylphosphorodiamidite (4.376 g, 14.52 mmol) was added, the reaction mixture was stirred for 16 h at RT. diluted to 300 ml with anhydrous DCM, the solid w^as filtered, and shaken with cold NaHCCb (150 mL) for 5 min. Organic layer was separated, washed with NaHCOs, dried (Na2SOr) and product was purified on CombiFlash® using 120 g SiO2 column. Eluent: hexane (A) - EtOAc (B), B= 10-50%, 60 min. Yield 6.29 g. Calculated: MW 1012.28. Found: MS (ES. positive): 1013.89 [M+H]+.Synthesis of RNAi Agents and TfRl-Specific ConjugatesF Conjugation of RNAi agents to Fabs and Capping with CP-1113.

[0267] RNAi agents described herein comprising a free amine were conjugated to L20-pusing standard amide reaction chemistry following cleavage from the solid phase. To a solution ofFab in PBS (0.2 umol, 1.0-10.0 mg / mL in PBS) was added a freshly prepared solution of (tris(2-carboxyethyl)phosphine) hydrochloride (TCEP-HC1) in PBS (5-20 eq, 70 mM). The reaction was held overnight at room temperature and covered from light. The next day. TCEP was removed by loading the reaction mixture on a PD-10 desalting column equilibrated with PBS and eluted with PBS. The concentration of Fab in the eluate was determined using the theoretical absorptivity factor at 280 nm. A solution of L20-modified sense strand in sodium phosphate buffer was prepared, and the concentration was determined using the theoretical absorptivity’ factor at 260 nm. To the desalted Fab solution was added L20-modified sense strand (1 -1.3 eq, 0.5-2.5 mM), and the reaction was mixed end- over-end. Analysis by SEC Method 1 and AIEX Method 1 show a mixture of starting Fab,DARI, and DAR2. After 1 hour, a solutionDMSO and added to the reaction mixture (3 eq, 36 mM). After 1 hour, a solution of L-cysteine in PBS was added to the reaction mixture (6-10 eq, 165 mM). Finally, the conjugate was annealed by addition of antisense strand (1 .2-1.5 eq, 0.5-2.5 mM). Tire conjugate was purified by an AKTA Pure FPLC system equipped with 20 mM tris pH 8 (Buffer A), 20 mM tris 1500 mM NaCl (Buffer B), and a 5 x 200 mm column packed with Tosoh SuperQ 5PW (20 micron). The crude reaction mixture was pump loaded onto the column and eluted with a gradient of 10- 40% Buffer B. DARI and DAR2 fractions were differentiated by SEC Method 1, AEX Method 1, and Nanodrop 260 / 280 readings. DARI fractions were pooled and buffer exchanged to PBS using a PD-10 desalting column. The purified conjugate was analyzed by SEC Method 1 and eluted as a monomeric peak with a retention time of 13.2 minutes.G. Conjugation ofRNAi agents to Fabs and Capping with NEM.

[0268] RNAi agents described herein comprising a free amine were conjugated to L20-pusing standard amide reaction chemistry following cleavage from the solid phase. To a solution of Fab in PBS (10 mg, 0.2 umol, 1.0-10.0 mg / mL in PBS) was added a freshly prepared solution of TCEP-HC1 in PBS (5-20 eq, 70 mM). The reaction was held overnight at room temperature and covered from light. The next day, TCEP was removed by loading the reaction mixture on to a PD-10 desalting column equilibrated with 20 mM tris pH 8 and eluted with 20 mM tris pH 8. The concentration of Fab in the eluate was determined using the theoretical absorptivity factor at 280 nm. A solution of L20-modified sense strand in sodium phosphate buffer was prepared, and the concentration was determined using the theoretical absorptivity factor at 260 nm. To the desalted Fab solution was added L20-modified sense strand (1 -1.3 eq, 0.5-2.5 mM), and the reaction was mixed end-over-end. Analysis by SEC Method 1 and AIEX Method 1 show a mixture of starting Fab, DARI , and DAR2. After 1 hour, a solution of N-ethyl maleimide (NEM) in 20 mM tris pH 8 was added to the reaction mixture (12 eq, 160 mM). After 1 h, the conjugate was annealed by addition of antisense strand (1.2-1.5 eq, 0.5-2.5 mM). The conjugate was purified by an AKTA Pure FPLC system equipped with 20 mM tris pH 8 (Buffer A), 20 mM tris 1500 mM NaCl (Buffer B), and a 5 x 200 mm column packed with Tosoh SuperQ 5PW (20 micron). The crude reaction mixture was loaded onto the column and eluted with a gradient of 10-40% Buffer B. DARI and DAR2 fractions were differentiated by SEC Method 1, AEX Method 1, and UV-Vis 260 / 280 measurements. DARI fractions were pooled and buffer exchanged to PBS using a PD-10 column. The purified conjugate was analyzed by SEC Method 1 and eluted as a monomeric peak with a retention time of 13.2 minutes.SEC Method 1AIEX Method 1

[0269] RNAi agents described herein comprising a free amine were conjugated to L1026-p:following cleavage from the solid phase according to the following procedure:

[0270] To a solution of Fab0070 (28 mg, 0.59 umol, 5.55 mg / mL in PBS) was added a freshly prepared solution of TCEP-HC1 in PBS (5 eq, 70 mM, 42 uL). The reduction was mixed endover-end at ambient temperature for 15 minutes then held at 5°C overnight without agitation. The next day, TCEP was removed by loading the reaction mixture on two PD-10 desaltingcolumns (Cytiva) equilibrated with 20 mM tris 50 mM NaCl pH 7.6 (alternatively, 20 mM tris pH 8 or PBS buffer can be used) and eluted with the same buffer. The concentration of the Fab in the eluate was determined using the theoretical absorptivity factor at 280 nm. A solution of L-1026-modified sense strand (CS915332) in 10 mM sodium phosphate buffer pH 6.0-6.5 was prepared, and the concentration was determined using the theoretical absorptivity factor at 260 nm. To the desalted Fab solution was added L-1026-modified CS915332 (1.15 eq, 2.75 mM, 240 uL), and the reaction was mixed end-over-end at ambient temperature. Analysis by SEC Method 1 and AIEX Method 1 show a mixture of starting Fab0070, DARI product, and DAR2 product. After 30 m, a solution of L-cysteine in 20 mM tris 50 mM NaCl pH 7.6 (alternatively, some L-1026 conjugates have been prepared in 20 mM tns pH 8 or PBS buffer solutions) was added to the reaction mixture (10 eq. 165 mM. 36 uL). After 30 m, the conjugate was annealed by addition of antisense strand (CA003820) (1.3 eq, 1.45 mM in water, 529 uL). The conjugate was purified by an AKTA Pure FPLC system equipped with 20 mM tris pH 8 (Buffer A), 20 mM tris 1500 mM NaCl (Buffer B), and a 5 x 200 mm column packed with Tosoh SuperQ 5PW (20 micron). The crude reaction mixture was loaded onto the column and eluted with a gradient of 10-40% Buffer B. DARI and DAR2 fractions were differentiated by SEC Method 1 , AIEX Method 1, and UV-Vis 260 / 280 measurements. DARI fractions were pooled and buffer exchanged to PBS using two PD-10 columns. The purified conjugate was analyzed by SEC Method 1 and eluted as a monomeric peak with a retention time of 7.2 minutes.SEC Method 1AIEX Method 1H. Conjugation of RNAi agents to Fabs.Synthesis of Fab0070-L-1288

[0271] To a solution of Fab0070 in PBS (82 mg, 8.2 mg / mL) was added a freshly prepared solution of TCEP-HC1 in PBS (5 eq). The reaction mixture was held overnight at 5°C. The next day, the reaction mixture was buffer exchanged to 20 mM tris 50 mM NaCl pH 7.6 using four PD-10 columns equilibrated with the same buffer. The concentration was determined by Nanodrop, and the solution was diluted to 2 mg / mL with 20 mM tris 50 mM NaCl pH 7.6. The solution was diluted with DMSO (1.95 mL). A solution of L-1288 was prepared (2 mg / mL in DMSO) and charged (1.6 mL, 3.19 mg, 3 eq) over 15 m. After 10 m, the reaction was quenched with cysteine (10 eq). Insoluble material was removed by centrifugation. The supernatant removed, filtered, diluted 1 : 1 with PBS, and concentrated / desalted using Pierce 10K MWCO spin columns to a volume of 8 mL. Finally, the solution was buffer exchanged by loading onto a HiPrep 26 / 10 desalting column and eluting with PBS. The final yield was approximately 80 mg.

[0272] Bioconjugation of L-1289

[0273] Lyophilized CS915332 (86 mg, 11.6 umol) was brought up in DMF (1.58 mL) and water (215 uL) and sonicated. To the solution was added triethylamine (9.7 uL, 6 eq) and a solution of L-1289 in DMF (50 mg / mL, 344 uL, 3 eq). The progress of the reaction w as monitored by LCMS. After 1 h, added additional triethylamine (4 eq) and L-1289 solution (2 eq). After 20 m, LCMS showed complete conversion. The reaction mixture was acidifiedwith phosphoric acid (200 mg / mL in water, 5 eq). The crude solution was added dropwise to a mixture of acetonitrile (43 mL) and PBS (1.7 mL). The precipitate was collected by centrifugation and the supernatant was discarded. The pellet was dissolved in water (1.5 mL) and added to acetonitrile (43 mL). The precipitate was collected by centrifugation and dissolved in PBS (5 mL). The yield was 88 mg (98%).

[0274] Conjugation of Fab0070-L-1288 with L-1307 modified sense strand

[0275] To a solution of Fab0070-L-1288 in PBS (12.5 mg, 6.97 mg / mL, 1.79 mL, 1 eq) was added a solution of L1307-modified sense strand:L-1307s(invAb)scacuuuugAfCfCfugcuaaucaas(invAb), (i.e., CS009529, SEQ ID NO.: 771) in PBS (6.18 mg, 17.9 mg / mL, 345 uL, 3 eq). The combined solution was mixed end-over- end for 10 m then stored at 5°C without agitation. The next day, the reaction was warmed to room temperature and annealed with antisense strand CA003820, 6.23 mg, 10.4 mg / mL, 599 uL, 3.3 eq. The conjugate w as purified by an AKTA Pure FPLC system equipped with 20 mM tris pH 8 (Buffer A), 20 mM tris 1500 mM NaCl (Buffer B), and a 5 x 200 mm column packed with Tosoh SuperQ 5PW (20 micron). The crude reaction mixture was diluted to 40 mL with MPA. loaded onto the column, and eluted with a gradient of 10-40% Buffer B. DARI -containing fractions were pooled and buffer exchanged to PBS using two PD-10 columns. The conjugate was analyzed by SEC Method 1 and found to be 99% pure with a retention time of 7.2 m. This procedure was also followed to generate the Fab-L-1288-L- 1289-RNAi conjugates disclosed herein.SEC Method 1I. Synthesis of antibody-siRNA linkers

[0276] The synthesis of various Fab linkers used throughout the present application are provided below.Synthesis of 2,3,5,6-tetrafluorophenyl 16-((3,5-bis(5-(methylsulfonyl)-l,3,4-oxadiazol-2- yl)pheiiyl)amino)-16-oxo-4,7,10,13-tetraoxahexadecanoate (i.e., L-1026-p)

[0277] Compound 6 (2.35 g, 7.31 mmol; prepared according to Sarbisheh et al. Bioconjugate Chemistry 2020 31 (12). 2789-2806), EDC-HC1 (2.38 g, 12.43 mmol), and K-Oxyma (2.50 g, 13.9 mmol) were combined as solids and slurried in DMF (190 mL) under N2 at ambient temperature. Compound 7 (1.1.92 g, 5.48 mmol) was added as a solution in DMF (10 mL). After 5 m, tri ethylamine (4.5 mL, 32.2 mmol) was added dropwise at ambient temperature. The reaction mixture was heated at 50°C for 2 days. The reaction mixture was concentrated under reduced pressure to a red oil which was slurried in DCM (250 mL) and washed with sat. aq. sodium bicarbonate (200 mL). The layers were separated, and the aqueous layer w?as further extracted with DCM (100 mL). The combined organic phase was washed with water (200 mL) and brine (200 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated. The residue was purified by normal phase SiCh chromatography with a gradient of ethyl acetate in DCM (0-100%). Yield of compound 8: 1.77 g (49%), partially contaminated with compound 6. Calculated mw for compound 8: 653.77 g / mol, found m / z (ESI, positive mode): 654.83.

[0278] Compound 8 (1.77 g, 2.71 mmol) w as dissolved in TFA:DCM [1 : 1 ] (18 mL) and stirred at ambient temperature for 1 hour. The reaction mixture was concentrated under reduced pressure then coevaporated with toluene (3 x 30 mL). The residue was purified by normal phase SiCh chromatography with a gradient of DCM containing 0.1% formic acid and methanol (0- 7%). Yield of compound 9: 1.30 g (80%). Calculated mw for compound 9: 597.66 g / mol, found m / z (ESI, positive mode): 598.79.

[0279] Compound 9 (1.30 g, 2.18 mmol) was dissolved in DCM (50 mL) and cooled to 0°C. A 100 mg / mL solution of m-CPBA solution was prepared by dissolving 10.38 g w-CPBA (77 wt%) in 80 mL DCM and drying with sodium sulfate until clear. To the solution of compound 9 was added 58 mL IM-CPBA (5.85 g, 26. 1 mmol) dropwise at 0°C. The reaction mixture was warmed to ambient temperature and allowed to proceed overnight. The reaction mixture was concentrated, slurried in DCM 0.1% formic acid (50 mL), and filtered. The filtrate was purified by normal phase SiCh chromatography with a gradient of DCM containing 0.1 % formic acid and methanol (0-10%). Yield of compound 10: 1.03 g (72%). Calculated mw for compound 10: 661.65 g / mol, found m / z (ESI, positive mode): 662.65.

[0280] To a solution of compound 10 (1.03 g, 1.56 mmol) in DCM:ACN [4: 11 (15 mL) at 0°C was added EDC (0.448 g, 2.34 mmol) followed by a solution of TFP (0.310 g, 1.87 mmol) in DCM:ACN [4: 1] (5 mL). After 5 m, the reaction mixture was warmed to ambient temperature. After 1.5 h, the reaction mixture was concentrated to dryness. The crude was purified by preparative reverse phase HPLC (Phenomenex Gemini Cl 8 50 mm x 250 mm, 10 urn) using a gradient of water / acetonitrile containing 0.1% TFA. Product-containing fractions were concentrated under reduced pressure. Yield of L-1026-p: 1.10 g (87%). Calculated mw for compound L-1026-p: 809.71 g / mol. found m / z (ESI, positive mode): 810.62.!H NMR (400 MHZ,[D6]DMSO, 25°C): 5 = 2.64 (t, 2H), 3.00 (t, 211 ). 3.49 (m, 12H), 3.74 (m, 10H), 7.92 (m, 1H), 8.34 (t, 1H), 8.68 (d, 2H), 10.67 (s, 1H).Synthesis of L20-p

[0281] To a suspension of compound 1 (5.00 g, 22.50 mmol) and CS2CO3 (25.66 g, 78.75 mmol) in anhydrous DMF (80 mL) was added methyl iodide (4.20 mL, 67.50 mmol) at room temperature. The reaction mixture was stirred at room temperature for 48 hours. The reaction mixture was quenched with water (200 mL) and the mixture was extracted with EtOAc (3 x 100 mL). The organic phase was combined and washed with w ater and brine. The organic layer was dried over anhydrous Na2SO4 and concentrated. Compound 2 was obtained as a light yellow solid. 5.41 g. 96%. Compound 2 was used directly without further purification. LC- MS: [M+H] calculated 251.05, found 251.1 8.

[0282] To a solution of compound 2 (5.41 g, 21.62 mmol) in THF7H2O (50 mL / 50 mL) was added LiOH (2.59 g, 108.08 mmol) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. After removing TFIF under vacuum, the pH was adjusted to ~2 by [C] HC1. Then EtOAc (3 x 60 mL) was used to extract. The organic lay ers were combined, washed with brine, then dried over anhydrous Na2.SO4, and concentrated. Compound 3 w7as obtained as an off-white solid, 5 g, 98%. Compound 3 was used directly without further purification. LC-MS: calculated [M+H] 237.03, found 237.26.

[0283] To a solution of compound 3 (5.81 g, 24.60 mmol) in THF / DMF (80 niL / 20 mL) was added EDC (7.07 g, 36.90 mmol), DMAP (0.30 g, 2.46 mmol) and compound 4 (6.13 g, 36.90 mmol) at room temperature. The reaction mixture was stirred at room temperature overnight. After removing solvent under vacuum, the residue was loaded on a 120 g column and compound 5 was eluted with 0-50% EtOAc in hexanes. Compound 5 was obtained as a white solid, 9.36 g, 99%. LC-MS: calculated [M+H] 385.03, found 385.46.

[0284] To a solution of compound 5 (2.29 g, 5.96 mmol) in DCM (1 10 mL) was added 70% m-CPBA (5.14 g, 27.79 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 6 hows. Another 1.8 g m-CPBA was added at room temperature. The reaction mixture was stirred at room temperature overnight. After filtration, the solvent was removed under vacuum. The residue was recrystallized from DCM / EtOAc (50 mL / 50 mL) twice. Compound L20-p was obtained as white needle cry stals, 1.93 g, 78%. LC-MS: calculated [M+H] 417, found 417.Example 2. In Vivo Knockdown of AR in Mice

[0285] AR RNAi agents were evaluated in vivo in mouse. On Day 1, four (n=4) female C57bl / 6 mice were administered, via subcutaneous (SC) injection, either saline or AR RNAi agents (formulated in saline, at 3.0 mg / kg animal body weight), at an injection volume of 10 mL / kg of animal weight. Dosing was in accordance with Table 12 below.

[0286] Table 12. Dosing groups for the mice of Example 2.

[0287] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included three N-acetyl- galactosamine groups (tridentate ligand) having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, and 7 for specific modifications and structure information related to the AR RNAi agents; see Table 11 for structure of (NAG37) and (NAG37)s ligand). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0288] On Day 8, the mice were euthanized. From the mice, liver (~50 mg) was harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 13.

[0289] Table 13. Relative expression of mAR in mice liver, of Example 2.

[0290] Groups 2-9 showed reduction in mAR out to at least Day 8. Most notably, a single dose of 3.0 mg / kg AC908478 achieved -79% inhibition (0.212).Example 3. In Vivo Knockdown of AR in Mice

[0291] AR RNAi agents were evaluated in vivo in mouse. On Day 1, four (n==4) female C57bl / 6 mice were administered, via intracerebroventricular (ICV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF. at 50 pg or 200 pg total RNAi agent), at a total injection volume of 50 pL. Dosing was in accordance with Table 14 below.

[0292] Table 14. Dosing groups for the mice of Example 3.

[0293] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included a lipid moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid and antigen binding moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0294] On Day 12, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPlA as endogenous gene, normalized to Group 1 mice dosed with aCSF. The mAR expression data is shown in the following Table 15.

[0295] Table 15. Relative expression of mAR in mice thoracic spinal cord and cerebellum, of Example 3.

[0296] In the thoracic spinal cord, Groups 3-5, 7, and 9 showed reduction in mAR out to at least Day 12. In the cerebellum. Groups 2-9 showed reduction in mAR out to at least Day 12. Most notably, a single dose 200 pg AC002146 achieved ~67% inhibition (0.331) in the cerebellum.Example 4. In Vivo Knockdown of AR in Mice

[0297] AR RNAi agents were evaluated in vivo in mouse. On Day 1, four (n=4) female C57bl / 6 mice were administered, via intracerebroventricular (ICV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF, at 100 pg total RNAi agent), at a total injection volume of 10 pL. Dosing was in accordance with Table 16 below'.

[0298] Table 16. Dosing groups for the mice of Example 4.

[0299] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 57terminal end of the sense strand to a targeting ligand that included a lipid moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7. 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0300] On Day 8, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with aCSF The mAR expression data is shown in the following Table 17.

[0301] Table 17. Relative expression of mAR in mice thoracic spinal cord, cerebellum, and cortex, of Example 4.

[0302] In the thoracic spinal cord. Groups 3, 6, 8, 9, and 11 showed reduction in niAR out to at least Day 8. In the cerebellum, Groups 2-9 and 11 showed reduction in mAR out to at least Day 8. In the cortex, Groups 2, 3, 6, and 8-11 showed reduction in mAR out to at least Day 8. Mostnotably, a single dose 100 pg AC909558 achieved -91% inhibition (0.086) in the thoracic spinal cord.Example 5. Synthesis In Vivo Knockdown of AR in Mice

[0303] AR RNAi agents were evaluated in vivo in mouse. On Day 1, three (n=3) female C57bl / 6 mice were administered, via intracerebroventricular (ICV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF, at 100 pg total RNAi agent), at a total injection volume of 10 pL. Dosing was in accordance with Table 18 below.

[0304] Table 18. Dosing groups for the mice of Example 5.

[0305] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5 ’ terminal end of the sense strand to a targeting ligand that included a lipid moiety- having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0306] On Day 8. the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with aCSF. The mAR expression data is shown in the following Table 19.

[0307] Table 19. Relative expression of mAR in mice thoracic spinal cord, cerebellum, and cortex, of Example 5.

[0308] In the thoracic spinal cord. Groups 2-13 showed reduction in mAR out to at least Day 8. In the cerebellum, Groups 3-5 and 7-13 showed reduction in mAR out to at least Day 8. In the cortex. Groups 3-13 showed reduction in mAR out to at least Day 8. Most notably, a single dose 100 pg AC909924 achieved -74% inhibition (0.256) in the thoracic spinal cord.Example 6. In Vivo Knockdown of AR in Mice.

[0309] AR RNAi agents were evaluated in vivo in mouse. On Day 1, three (n=3) female C57bl / 6 mice were administered, via intracerebroventricular (ICV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF. at 100 pg total RNAi agent), at a total injection volume of 10 pL. Dosing was in accordance with Table 20 below.

[0310] Table 20. Dosing groups for the mice of Example 6.

[0311] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included a lipid moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0312] On Day 8, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPlA as endogenous gene, normalized to Group 1 mice dosed with aCSF. The tnAR expression data is shown in the following Table 21.

[0313] Table 21. Relative expression of mAR in mice thoracic spinal cord and cerebellum, of Example 6.

[0314] In the thoracic spinal cord. Groups 2-20 showed reduction in mAR out to at least Day 8. In the cerebellum, Groups 2-15 and 17-20 showed reduction in mAR out to at least Day 8. Most notably, a single dose 100 pg AC909896 achieved -89% inhibition (0. 113) in the thoracic spinal cord.Example 7. In Vivo Knockdown of AR in Mice

[0315] AR RNAi agents were evaluated In vivo in mouse. On Day 1, four (n=4) female C57bl / 6 mice were administered, via intracerebroventricular (ICV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF. at 70 pg total RNAi agent), at a total injection volume of 10 pL. Dosing was in accordance with Table 22 below.

[0316] Table 22. Dosing groups for the mice of Example 7.

[0317] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5 ’ terminal end of the sense strand to a targeting ligand that included a lipid moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0318] On Day 8, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, and cortex were harvested and collected for analysis. mAR mRNA transcriptexpression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with aCSF. The mAR expression data is shown in the following Table 23.

[0319] Table 23. Relative expression of mAR in mice thoracic spinal cord, cerebellum, and cortex, of Example 7.

[0320] In the thoracic spinal cord, cerebellum, and cortex. Groups 2-16 showed reduction in mAR out to at least Day 8. Most notably, a single dose 70 pg AC910174 achieved -96% inhibition (0.040) in the cerebellum.Example 8. In Vivo Knockdown of AR in Mice

[0321] AR RNAi agents were evaluated in vivo in mouse. On Day 1, four (n=4) female C57bl / 6 mice were administered, via intracerebroventricular (ICV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF. at 100 pg total RNAi agent), at a total injection volume of 10 pL. Dosing was in accordance with Table 24 below.

[0322] Table 24. Dosing groups for the mice of Example 8.

[0323] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included a lipid moiety ha ving the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0324] On Day 8, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with aCSF. The mAR expression data is shown in the following Table 25.

[0325] Table 25. Relative expression of mAR in mice thoracic spinal cord, cerebellum, and cortex, of Example 8.

[0326] In the thoracic spinal cord, Groups 3-15 showed reduction in mAR out to at least Day 8. In the cerebellum and cortex, Groups 2-15 showed reduction in mAR out to at least Day 8. Most notably, a single dose 100 ug AC910278 achieved -71% inhibition (0.285) in the cerebellum.Example 9. In Vivo Knockdown of AR in Mice

[0327] AR RNAi agents were evaluated in vivo in mouse. On Day 1, four (n=4) female C57bl / 6 mice were administered, via intracerebroventricular (1CV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF, at 70 pg total RNAi agent), at a total injection volume of 10 pL. Dosing was in accordance with Table 26 below.

[0328] Table 26. Dosing groups for the mice of Example 9.

[0329] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5 ’ terminal end of the sense strand to a targeting ligand that included a lipid moiety- having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0330] On Day 8. the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with aCSF. The mAR expression data is shown in the following Table 27.

[0331] Table 27. Relative expression of mAR in mice thoracic spinal cord, cerebellum, and cortex, of Example 9.

[0332] In the thoracic spinal cord, cerebellum, and cortex, Groups 2-12 showed reduction in mAR out to at least Day 8. Most notably, a single dose 70 ug AC910287 achieved -84% inhibition (0. 159) in the thoracic spinal cord.Example 10. In Vivo Knockdown of AR in Mice

[0333] AR RNAi agents were evaluated in vivo in mouse. On Day 1, four (n=4) female C57H / 6 mice were administered, via intracerebroventricular (ICV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF, at 30 pg, 60 pg, or 120 pg total RNAi agent), at a total injection volume of 10 pL. Dosing was m accordance with Table 28 below.

[0334] Table 28. Dosing groups for the mice of Example 10.

[0335] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included a lipid moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0336] On Day 8, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with aCSF. The mAR expression data is shown in the following Table 29.

[0337] Table 29. Relative expression of mAR in mice thoracic spinal cord, cerebellum, and cortex, of Example 10.In the thoracic spinal cord and cerebellum, Groups 2-19 showed reduction in mAR out to at least Day 8. In the cortex, Groups 2, 3, and 7-19 showed reduction in mAR out to at least Day 8. Most notably, a single dose 120 ug AC910174 achieved -91% inhibition (0.090) in the thoracic spinal cord.Example 11. In Vivo Knockdown of AR in Mice

[0338] AR RNAi agents were evaluated in vivo in mouse. On Day 1 , four (n=4) female C57bl / 6 mice were administered, via intracerebroventricular (ICV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF, at 3 pg, 10 pg, 30 pg, 90 pg, or 180 pg total RNAi agent), at a total injection volume of 10 pL. Dosing was in accordance with Table 30 below-.

[0339] Table 30. Dosing groups for the mice of Example 11.

[0340] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included a lipid moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and10 for specific modifications and structure information related to the AR RNAi agents; including the lipid moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0341] On Day 8, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with aCSF. The mAR expression data is shown in the following Table 31.

[0342] Table 31. Relative expression of mAR in mice thoracic spinal cord, cerebellum, and cortex, of Example 11.

[0343] In the thoracic spinal cord and cortex. Groups 3-6 showed reduction in mAR out to at least Day 8. In the cerebellum, Groups 2-6 showed reduction in mAR out to at least Day 8. Most notably, a single dose 180 ug AC911150 achieved -86% inhibition (0.135) in the thoracic spinal cord. In the thoracic spinal cord, cerebellum, and cortex, a dose-response was observed for AC911150.Example 12. In Vivo Knockdown of AR in Mice

[0344] AR RNAi agents were evaluated in vivo in mouse. On Day 1, four (n=4) female C57bl / 6 mice were administered, via intracerebroventricular (ICV) injection, either artificial cerebrospinal fluid (aCSF) or AR RNAi agents (formulated in aCSF, at 3 pg, 10 pg, 30 pg, 90 pg, or 180 pg total RNAi agent), at a total injection volume of 10 pL. Dosing was in accordance with Table 32 below.

[0345] Table 32. Dosing groups for the mice of Example 12.

[0346] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5' terminal end of the sense strand to a targeting ligand that included a lipid moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0347] On Day 8, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with aCSF The mAR expression data is shown in the following Table 33.

[0348] Table 33. Relative expression of mAR in mice thoracic spinal cord, cerebellum, and cortex, of Example 12.

[0349] In the thoracic spinal cord and cortex, Groups 3-6 showed reduction in mAR out to at least Day 8. In the cerebellum, Groups 2-6 showed reduction in mAR out to at least Day 8. Most notably, a single dose 180 ug AC911151 achieved -89% inhibition (0.106) in the cerebellum.Example 13. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0350] AR RNAi agents were evaluated in vivo in mouse. On Day 1 and Day 2. five (n=5) female B-hTFRl mice were administered, via intravenous (IV) injection, either saline or AR RNAi agents (formulated in saline at 1.5 mg / kg animal body weight), at total dosing volume of 250 gL / 25g. Dosing was in accordance with Table 34 below.

[0351] Table 34. Dosing groups for the mice of Example 13.

[0352] B-hTFRl mice, also known as C57BL / 6-Tfrctml(TERC)Bcgen / Bcgen mice (Biocytogen), have the exons 4-19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0353] Each of the AR RN Ai agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3. 4, 5, 6, 7, 8. 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0354] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 35.

[0355] Table 35. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 13.

[0356] In the thoracic spinal cord, cerebellum, cortex, and striatum. Groups 2-7 showed reduction in mAR out to at least Day 15. Most notably, two doses of 1.5 mg / kg AC003313 achieved ~87% inhibition (0.127) in the cortex.Example 14. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0357] AR RNAi agents were evaluated m vivo in mouse. On Day 1 and Day 2, five (n=5) female B-hTFRl mice were administered, via intravenous (IV) injection, either saline or AR RNAi agents (formulated in saline at 1.5 mg / kg animal body weight), at total dosing volume of 250 pL / 25g. Dosing was in accordance with Table 36 below.

[0358] Table 36. Dosing groups for the mice of Example 14.

[0359] B-hTFRl mice, also known as C57BL / 6-Tfrctml(TFRC)Bcgen / Bcgen mice (Biocytogen), have the exons 4-19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0360] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties). The AR RNAi agents evaluated in thisExample study were cross-reactive across mouse and human AR.

[0361] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 37.

[0362] Table 37. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 14.

[0363] In the thoracic spinal cord, cerebellum, cortex, and striatum. Groups 2-10 showed reduction in mAR out to at least Day 15. Most notably, two doses of 1.5 mg / kg AC004798 achieved -86% inhibition (0.140) in the cerebellum.Example 15. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0364] AR RNAi agents were evaluated in vivo in mouse. On Day 1 and Day 2. five (n=5) female B-hTFRl mice were administered, via intravenous (IV) injection, either saline or AR RNAi agents (formulated in saline at 1.5 mg / kg animal body weight), at total dosing volume of 250 pL / 25g. Dosing was in accordance with Table 38 below.

[0365] Table 38. Dosing groups for the mice of Example 15.

[0366] B-hTFRl mice, also known as C57BL / 6-Tfrctml(TFRC)Bcgen / Bcgen mice (Biocytogen), have the exons 4- 19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0367] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3. 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0368] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 39.

[0369] Table 39. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 15.

[0370] In the thoracic spinal cord, cerebellum, cortex, and striatum, Groups 2-6 showed reduction in mAR out to at least Day 15. Most notably, two doses of 1.5 mg / kg AC004997 achieved -80% inhibition (0.203) in the cerebellum.Example 16. In Vivo Knockdown of AR mRNA gene expression in Cynomolgus Monkeys.

[0371] AR RNAi agents were evaluated in vivo in Cynomolgus monkeys. On Day 0 and Day 7, three (n=3) male Cynomolgus monkeys for each test group were dosed with AR RNAi agents formulated in PBS at 3.0 mg / kg (adjusted for individual animal body weight), 2,0 mL / kg dose volume, at 1.5 mg / ml dose concentration, or dosed with PBS. Each dose was administered via subcutaneous (SC) injection, and each dose was administered based on each respective animal's most recent body weight. The dosing was in accordance with the following Table 40.

[0372] Table 40. Dosing for Cynomolgus monkeys of Example 16.

[0373] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3. 4, 5. 6. 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across cyno and human AR.

[0374] The test animals were of non-human primate (NHP), Cynomolgus monkeys, male, naive, ~3-7 years of age, body weights ~3.0-8.0 kg. All NHP housing and research procedures conform to U.S. Department of Agriculture’s (USDA) Animal Welfare Act.

[0375] Each cyno from Groups 1-4 were dosed on Day 0 and 7. The RNAi agent test articles were administered via subcutaneous (SC) administration. The SC administration sites were marked and recorded, and SC injection site on Day 0 did not overlap with the injection site of Day 7. The dose volumes were calculated from NHP body weights collected prior to dose. Weights were measured prior to dosing for the calculation of dose volumes, then once weekly. More frequent body weight measurements were collected if animal becomes sick.

[0376] At Day 28. the Cynomolgus monkeys were euthanized. From the test animals, the following tissues were collected: left and right brain hemisphere, spinal cord, dorsal root ganglion (DRG). Tissues other than the aforementioned tissues may also be collected, and should such extra tissues be collected, their biological data were similarly presented below; The collected tissues were analyzed for biological parameters.

[0377] cAR mRNA transcript expression w^as analyzed via qPCR in the cyno tissues, with cPPIB as endogenous control gene, normalized to Group 1 cynos dosed with PBS. The cAR mRNA transcript expression data is shown in the following Table 41.

[0378] Table 41. Relative expression of cAR in thoracic spinal cord, cerebellum, striatum, and cortex, of Example 16.

[0379] As shown in Table 41, above, durable reduction of ARmRNA expression was observed in multiple tissues for non-human pnmates treated with anti-TfRl antibody-siRNA conjugate. Groups 3 (Fab0061 -siRNA conjugate) and 4 (Fab0070-siRNA conjugate) outperformed Group 2 (Fab0002-siRNA conjugate) in nearly every tissue analyzed, with the exception of Thoracic DRG and Lumbar DRG.Example 17. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0380] AR RNAi agents were evaluated in vivo in mouse. On Day 1, four (n=4) female B- hTFRl mice were administered, via subcutaneous (SC) injection, either saline or AR RNAi agents (formulated in saline at 0.5, 1.5 or 4.5 mg / kg animal body weight), at total dosing volume of 250 p.L / 25g. Dosing was in accordance with Table 42 below.

[0381] Table 42. Dosing groups for the mice of Example 17.

[0382] B-hTFRl mice, also known as C57BL / 6-Tfrctml(TFRC)Bcgen / Bcgen mice (Biocytogen), have the exons 4-19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0383] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3. 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties).

[0384] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table

[0385] Table 43. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 17.

[0386] In the cortex and cerebellum, Groups 2-13 showed reduction in mAR out to at least Day 15. In the thoracic spinal cord. Groups 2-7, 9. 10. and 13 showed reduction in mAR out to at least Day 15. In the striatum, Groups 3-10 and 13 showed reduction in mAR out to at least Day 15. Most notably, one dose of 4.5 mg / kg AC004798 achieved -90% inhibition (0 100) in the cerebellum. In the thoracic spinal cord and cerebellum, a dose-response was observed for AC003313. AC004791, AC004792. and AC004798. In the cortex, a dose-response was observ ed for AC003313, AC004791, and AC004792. In the striatum, a dose-response was observed for AC004791, AC004792, and AC004798.Example 18. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0387] AR RNAi agents were evaluated in vivo in mouse. On Day 1 and Day 2, five (n=5) female B-hTFRl mice were administered, via subcutaneous (SC) injection, either saline or AR RNAi agents (formulated in saline at 0.75 or 1.5 mg / kg animal body weight), at total dosing volume of 250 pL / 25g. Dosing was in accordance with Table 44 below.

[0388] Table 44. Dosing groups for the mice of Example 18.

[0389] B-hTFRl mice, also known as C57BL / 6-Tfrctml(TFRC)Bcgen / Bcgen mice (Biocytogen), have the exons 4- 19 of mouse Tfrl gene that encode the extracellular region replaced by humanTFR1 exons 4-19.

[0390] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5 ' terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties).

[0391] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 45.

[0392] Table 45. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 18.

[0393] In the thoracic spinal cord, right cerebellum, right cortex, and right striatum. Groups 2- 12 showed reduction in mAR out to at least Day 15. Most notably, two doses of 1.5 mg / kg AC005488 achieved -92% inhibition (0.076) in the thoracic spinal cord. In the thoracic spinal cord right cerebellum, right cortex, and right striatum, a dose-response was observed for AC004791, AC004792, and AC004798.Example 19. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0394] AR RNAi agents were evaluated in vivo in mouse. On Day 1 and Day 2, five (n=5) female B-hTFRl mice were administered, via subcutaneous (SC) injection, either saline or AR RNAi agents (formulated in saline at 1.5 mg / kg animal body weight), at dose concentration 0. 15 mg / mL and 10 mL / kg dose volume. Dosing was in accordance with Table 46 below.

[0395] Table 46. Dosing groups for the mice of Example 19.

[0396] B-hTFRl mice, also known as C57BL / 6-Tfrctal ,(IFRC,Bcgen / Bcgen mice (Biocytogen). have the exons 4-19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0397] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5?terminal end of the sense strand to a targeting ligand that included an antigen binding moiety har ing the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5,6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moi eties).

[0398] On Day 15, the mice were euthanized. From the mice, CNS and muscle tissues were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. Tlie mAR expression data is shown in the following Table 47.

[0399] Table 47. Relative expression of mAR in mice CNS and muscle tissues, of Example 19.

[0400] In the cerebellum, cortex, triceps, gastrocnemius, thoracic spinal cord, and right striatum. Groups 2 and 3 showed reduction in mAR out to at least Day 15. Most notably, two doses of 1.5 mg / kg AC011951 achieved -84% inhibition (0. 157) in the cerebellum.Example 20. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0401] AR RNAi agents were evaluated in vivo in mouse. On Day 1 and Day 2. five (n=5) female B-hTFRl mice were administered, via subcutaneous (SC) injection, either saline or ARRNAi agents (formulated in saline at 0.5 or 1.5 mg / kg animal body weight), at total dosing volume of 250 pL / 25g. Dosing was in accordance with Table 48 below.

[0402] Table 48. Dosing groups for the mice of Example 20.

[0403] B-hTFRl mice, also known as C57BL / 6-Tfrctal l( IFRC )Bcgen / Bcgen mice (Biocytogen), have the exons 4-19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0404] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5 ’ terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties).

[0405] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 49.

[0406] Table 49. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 20.

[0407] In the thoracic spinal cord and cerebellum. Groups 2- 11 showed reduction in mAR out to at least Day 15. In the cortex and striatum. Groups 2-7, 9. and 11 showed reduction in mAR out to at least Day 15. Most notably, two doses of 1.5 mg / kg AC004793 achieved -83% inhibition (0.172) in the cerebellum In the thoracic spinal cord and cortex, a dose-response was observed for AC003313. AC004793, AC004799. AC004800, and AC004998. In the cerebellum and striatum, a dose-response was observed for AC003313, AC004793, AC004800, and AC004998.Example 21. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0408] AR RNAi agents were evaluated in vivo in mouse. On Day 1 and Day 2, five (n=5) female B-hTFRl mice were administered, via subcutaneous (SC) injection, either saline or AR RNAi agents (formulated in saline at 1.5 mg / kg animal body weight), at total dosing volume of 250 pL / 25g. Dosing was in accordance with Table 50 below.

[0409] Table 50. Dosing groups for the mice of Example 21.

[0410] B-hTFRl mice, also known as C57BL / 6-Tfrctol l(IFRC,Bc®en / Bcgen mice (Biocytogen), have the exons 4-19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0411] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5?terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6. 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties).

[0412] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 51.

[0413] Table 51. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 21.

[0414] In the thoracic spinal cord, cerebellum, cortex, and striatum. Groups 2-10 showed reduction in mAR out to at least Day 15. Most notably, two doses of 1.5 mg / kg AC006266 achieved -81 % inhibition (0.192) in the cerebellum.Example 22. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0415] AR RNAi agents were evaluated in vivo in mouse. On Day 1. Day 1 & 2. or Day 1 &2 & 3 & 4, five (n=5) female B-hTFRl mice were administered, via subcutaneous (SC) injection, either saline or AR RNAi agents (formulated in saline at 0.75, 1.5, 3.0, and 12.0 mg / kg animal body weight), at total dosing volume of 250 pL / 25g. Dosing was in accordance with Table 52 below.

[0416] Table 52. Dosing groups for the mice of Example 22.

[0417] B-hTFRl mice, also known as C57BL / 6-Tfrctol l(IFRC,Bc®en / Bcgen mice (Biocytogen), have the exons 4-19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0418] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5?terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6. 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties).

[0419] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with rnPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 53.

[0420] Table 53. Relative expression of mAR in mice thoracic spinal cord, cerebellum. striatum, and cortex, of Example 22.

[0421] In the thoracic spinal cord, cerebellum, cortex, striatum, and mid-brain, Groups 2-10 showed reduction in mAR out to at least Day 15. Most notably, four doses of 3.0 mg / kg AC006372 achieved ~85% inhibition (0. 153) in the cerebellum. In the thoracic spinal cord and cerebellum, a dose-response was observed for the single dose, two doses, and four doses of AC006372. In the cortex, a dose-response was observed for the single dose and two doses of AC006372. In the striatum, a dose-response was observed for the single dose and the four doses of AC006372. In the mid-brain, a dose-response was observed for the two doses and four doses of AC006372.Example 23. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0422] AR RNAi agents were evaluated in vivo in mouse. On Day 1 and Day 2. five (n=5) female B-hTFRl mice were administered, via subcutaneous (SC) injection, either saline or AR RNAi agents (formulated in saline at 1.5 mg / kg animal body weight), at total dosing volume of 250 gL / 25g. Dosing was in accordance with Table 54 below.

[0423] Table 54. Dosing groups for the mice of Example 23.

[0424] B-hTFRl mice, also known as C57BL / 6-Tfrctml(TFRC)Bcge7Bcgen mice (Biocytogen), have the exons 4-19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0425] Each of the AR RNAi agents included modified nucleotides that were conj ugated at the 5?terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3. 4. 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moi eties).

[0426] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. Additionally, muscle tissues were also collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 55.

[0427] Table 55. Relative expression of mAR in mice CNS and muscle tissues, of Example 23.

[0428] In both the CNS and muscle tissues, Groups 2-7 showed reduction in mAR out to at least Day 15. Most notably, two doses of 1 5 mg / kg AC009278 achieved -80% inhibition (0.179) in the right cerebellum, and two doses of 1.5 mg / kg AC009279 achieved -74% inhibition (0.259) in the right gastrocnemius.Example 24. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0429] AR RNAi agents were evaluated in vivo in mouse. On Day 1 and Day 2. five (n=5) female B-hTFRl mice were administered, via subcutaneous (SC) injection, either saline or AR RNAi agents (formulated in saline at 1.5 mg / kg animal body weight), at total dosing volume of 250 [iL / 25g. Dosing was in accordance with Table 56 below.

[0430] Table 56. Dosing groups for the mice of Example 24.

[0431] B-hTFRl mice, also known as C57BL / 6-Tfrctal(TFRC,Bc8C1VBcgen mice (Biocytogen), have the exons 4- 19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0432] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that in cl uded an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties).

[0433] On Day 15, the mice were euthanized. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene, normalized to Group 1 mice dosed with saline. The mAR expression data is shown in the following Table 57.

[0434] Table 57. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 24.

[0435] In the thoracic spinal cord, cerebellum, cortex, and striatum, Groups 2-7 showed reduction in mAR out to at least Day 15. Most notably, two doses of 1.5 mg / kg AC006895 achieved -76% inhibition (0.237) in the cerebellum.Example 25. In Vivo Knockdown of AR in Transgenic Tg B-hTfrl Mice.

[0436] AR RNAi agents were evaluated in vivo in mouse. On Day 1, Day 2, and Day 3, four (n=4) female B-hTFRl mice were administered, via subcutaneous (SC) injection, either saline or AR RNAi agents (formulated in saline at 3.0 mg / kg animal body weight), at total dosing volume of 250 pL / 25g. Dosing was in accordance with Table 58 below.

[0437] Table 58. Dosing groups for the mice of Example 25.

[0438] B-hTFRl mice, also known as C57BL / 6-Tfrctml(TFRC)Bcgen / Bcgen mice (Biocytogen), have the exons 4-19 of mouse Tfrl gene that encode the extracellular region replaced by human TFR1 exons 4-19.

[0439] Each of the AR RN Ai agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3. 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties).

[0440] On Day 4, 10, 17, 31, 45, 59, 80, 100, or 122, the mice were euthanized in accordance with Table 58 above. From the mice, thoracic spinal cord and cerebellum, striatum, and cortex were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mPPIA as endogenous gene; Groups 3-7 were normalized to Group 1 mice dosed with saline, Groups 9-11 were normalized to Group 8 mice dosed with saline. The mAR expression data is shown in the following Table 59A and Table 59B.

[0441] Table 59A. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 25.

[0442] In the thoracic spinal cord, cerebellum, left cortex, striatum, and brain stem, Groups 3- 7 showed reduction in mAR out to at least Day 59. Most notably, three doses of 3.0 mg / kg AC006448 achieved -85% inhibition (0.147) in the cerebellum at Day 59.

[0443] Table 59B. Relative expression of mAR in mice thoracic spinal cord, cerebellum, striatum, and cortex, of Example 25.

[0444] In the thoracic spinal cord, cerebellum, and striatum, Groups 9-11 showed reduction in mAR out to at least Day 122. Most notably, three doses of 3.0 mg / kg AC006448 achieved -39% inhibition (0.612) in the striatum at Day 122.Example 26. In Vivo Knockdown of AR in Rats.

[0445] AR RNAi agents were evaluated in vivo in rat. On Day 1, four (n=4) male Sprague- Dawley mice were administered, via intravenous (IV) injection, either saline or AR RNAi agents (formulated in saline at 1.5 mg / kg animal body weight), at total dosing volume of 250 pL / 25g. Dosing was in accordance with Table 60 below.

[0446] Table 60. Dosing groups for the rats of Example 26.

[0447] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5’ terminal end of the sense strand to a targeting ligand that included an antigen binding moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3. 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the antigen binding moieties).

[0448] On Day 15, the rats were euthanized. From the rats, muscle tissues were harvested and collected for analysis. rAR mRNA transcript expression was analyzed via qPCR, withrGAPDH as endogenous gene, normalized to Group 1 rats dosed with saline. The rAR expression data is shown in the following Table 61 .

[0449] Table 61. Relative expression of rAR in rat CNS and muscle tissues, of Example 26.

[0450] In the muscle tissues. Groups 2-5 showed reduction in rAR out to at least Day 15. Most notably, one dose of 3.0 mg / kg AC003039 achieved -90% inhibition (0.096) in the diaphragm.Example 27. In Vivo Knockdown of AR in Mice.

[0451] AR RNAi agents were evaluated in vivo in mouse. On Day 1, four (n=4) female C57bl / 6 mice were administered, via intravenous (IV) injection, either saline or AR RNAiagents (formulated in saline, at 3.0 mg / kg animal body weight), at total dosing volume of 200 pL / '20g. Dosing was in accordance with Table 62 below.

[0452] Table 62. Dosing groups for the mice of Example 27.

[0453] Each of the AR RNAi agents included modified nucleotides that were conjugated at the 5' terminal end of the sense strand to a targeting ligand that included a lipid moiety having the modified sequences as set forth in the duplex structures herein (see Tables 3, 4, 5, 6, 7, 8, 9 and 10 for specific modifications and structure information related to the AR RNAi agents; including the lipid moieties). The AR RNAi agents evaluated in this Example study were cross-reactive across mouse and human AR.

[0454] In accordance with Table 62 above, on Day 3, 29, 57, 85, 113, or 141, the mice were euthanized. From the mice, muscle tissues (triceps and gastrocnemius) were harvested and collected for analysis. mAR mRNA transcript expression was analyzed via qPCR, with mARLl as endogenous gene; each Group dosed with AC003039 was normalized to its respective saline-dosed Group that was sacrificed on the same day (i.e.. Groups 7 and 8 normalized to Group 1, Group 9 normalized to Group 2, Group 10 normalized to Group 3, Group 11 normalized to Group 4, Group 12 normalized to Group 5, and Group 13 normalized to Group 6). The mAR expression data is shown in the following Table 63.

[0455] Table 63. Relative expression of mAR in mice triceps and gastrocnemius, of Example 27.

[0456] In the triceps and gastrocnemius, Groups 7-13 showed reduction in mAR out to at least Day 141. Most notably, a single dose 3.0 mg / kg AC003039 achieved -72% inhibition (0.283) in the gastrocnemius on Day 141 .OTHER EMBODIMENTS

[0457] It is to be understood that while the invention has been described in conj unction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

CLAIMSWhat is claimed is:

1. An RNAi agent for inhibiting expression of an Androgen Receptor (AR) gene, comprising: an antisense strand comprising at least 17 contiguous nucleotides differing by 0 or 1 nucleotides from any one of the sequences provided in Table 2 or Table 3; and a sense strand comprising a nucleotide sequence that is at least partially complementary to the antisense strand.

2. The RNAi agent of claim 1, wherein the antisense strand comprises nucleotides 2-18 of any one of the sequences provided in Table 2 or Table 3.

3. The RNAi agent of claim 1 or claim 2, wherein the sense strand comprises a nucleotide sequence of at least 17 contiguous nucleotides differing by 0 or 1 nucleotides from any one of the sequences provided in Table 2 or Table 4, and wherein the sense strand has a region of at least 85% complementarity over the 17 contiguous nucleotides to the antisense strand.

4. The RNAi agent of any one of claims 1-3. wherein at least one nucleotide of the AR RNAi agent is a modified nucleotide or includes a modified intemucleoside linkage.

5. The RNAi agent of any one of claims 1-4, wherein all or substantially all of the nucleotides are modified nucleotides.

6. The RNAi agent of any one of claims 4-5, wherein the modified nucleotide is selected from the group consisting of: 2'-O-methyl nucleotide, 2’-fluoro nucleotide, 2'-deoxy nucleotide, 2’,3'-seco nucleotide mimic, locked nucleotide, 2'-F-arabino nucleotide, 2'- methoxy ethyl nucleotide, abasic nucleotide, ribitol, inverted nucleotide, inverted 2'-O-methyl nucleotide, inverted 2'-deoxy nucleotide, 2'-amino-modified nucleotide, 2'-alkyl-modified nucleotide, morpholino nucleotide, vinyl phosphonate-containing nucleotide, cyclopropyl phosphonate-containing nucleotide, and 3'-O-methyl nucleotide.

7. The RNAi agent of claim 5, wherein all or substantially all of the nucleotides are modified with 2'-O-methyl nucleotides, 2'-fluoro nucleotides, or combinations thereof.

8. The RNAi agent of any one of claims 1-7, wherein the antisense strand comprises the nucleotide sequence of any one of the modified sequences provided in Table 3.

9. The RNAi agent of any one of claims 1-8. wherein the sense strand comprises the nucleotide sequence of any one of the modified sequences provided in Table 4.

10. The RNAi agent of claim 1, wherein the antisense strand comprises the nucleotide sequence of any one of the modified sequences provided in Table 3 and the sense strand comprises the nucleotide sequence of any one of the modified sequences provided in Table 4.

11. The RNAi agent of any one of claims 1-10, wherein the sense strand is between 18 and 30 nucleotides in length, and the antisense strand is between 18 and 30 nucleotides in length.

12. The RNAi agent of claim 11, wherein the sense strand and the antisense strand are each between 18 and 27 nucleotides in length.

13. The RNAi agent of claim 12, wherein the sense strand and the antisense strand are each between 18 and 24 nucleotides in length.

14. The RNAi agent of claim 13, wherein the sense strand and the antisense strand are each 21 nucleotides in length.

15. The RNAi agent of claim 14, wherein the RNAi agent has two blunt ends.

16. The RNAi agent of any one of claims 1-15, wherein the sense strand comprises one or two terminal caps.

17. The RNAi agent of any one of claims 1-16, wherein the sense strand comprises one or two inverted abasic residues.

18. The RNAi agent of claim 1, wherein the RNAi agent comprises of a sense strand and an antisense strand that form a duplex having the structure of any one of the duplexes in Table 7, Table 8, Table 9, or Table 10.

19. The RNAi agent of claim 18, wherein all or substantially all of the nucleotides are modified nucleotides.

20. The RNAi agent of claim 19, wherein all or substantially all of the nucleotides are modified nucleotides.

21. The RNAi agent of any one of claims 19-20, wherein the sense strand further includes inverted abasic residues at the 3’ terminal end of the nucleotide sequence, at the 5’ end of the nucleotide sequence, or at both.

22. The RNAi agent of any one of claims 1-21, wherein the RNAi agent is linked to a lipid moiety or linked to an antigen binding protein.

23. The RNAi agent of any one of claims 1-22, wherein the RNAi agent is linked to a lipid moiety.

24. The RNAi agent of claim 23, wherein the lipid moiety is selected from the groupwherein ? indicates the point of connection to the RNAi agent.

25. The RNAi agent of claim 23 or claim 24, wherein the lipid moiety is conjugated to the sense strand.

26. The RNAi agent of claim 25, wherein the lipid moiety is conjugated to the 5’ terminal end of the sense strand.

27. The RNAi agent of any of claims 1-22, wherein the RNAi agent is linked to an antigen binding protein.

28. The RNAi agent of any one of claims 1-27, wherein the antigen binding protein is an antibody fragment (Fab) that specifically binds to one or more epitopes on a transferrin receptor (TfRl).

29. The RNAi agent of claim 28, wherein the Fab comprises (i) 6 complementary determining regions (CDRs), (ii) 3 CDRs on the variable light chain (VL), and / or (iii) 3 CDRs on the variable heavy chain (VH).

30. The RNAi agent of claim 29, wherein the variable light chain has a VL CDR1 sequence selected from the group consisting of: RASDGLYSNLA (SEQ ID NO: 721). RASDNLYRNLA (SEQ ID NO: 722), and RASDKLYSNLA (SEQ ID NO: 723); a VL CDR2 sequence selected from the group consisting of: DATLLAS (SEQ ID NO: 724), DARNLAS (SEQ ID NO: 725), DAFNLAS (SEQ ID NO: 726), DATRLAS (SEQ ID NO: 727). DATKLAS (SEQ ID NO: 728), and DAKNLAS (SEQ ID NO: 729); and / or a VL CDR 3 sequence of QHFWGTPLT (SEQ ID NO: 730).

31. The RNAi agent of claim 29 or 30, wherein the variable light chain comprises the sequence:DIQLTQSPSSLSASVGDRVTITCRASDKLYSNLAWYQQKPGKAPKLLIYDATLLASGV PSRFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGQGTKVEIK (SEQ ID NO: 747).

32. The RNAi agent of any one of claims 29-31, wherein the variable heavy chain has a VH CDR1 sequence selected from the group consisting of: GYTFNSYWMH (SEQ ID NO: 731), GYTFKSYWMH (SEQ ID NO: 732), GFTFTSYWMH (SEQ ID NO: 733), GYTFTSYWVH (SEQ ID NO: 734), and GYTFTSYWMH (SEQ ID NO: 735), a VH CDR2 sequence selected from the group consisting of: EINPTNGRVNYIEKFKS (SEQ ID NO: 736), EINPTNGRFNYIEKFKS (SEQ ID NO: 737), EINPTNGRTNYIEKFKS (SEQ ID NO:738), and EINPTNGRSNYIEKFKS (SEQ ID NO: 739); and / or a VH CDR3 sequence of: GTRAYHY (SEQ ID NO: 740).

33. The RNAi agent of any one of claims 29-32, wherein the variable heavy chain comprises the sequence:EVQLVESGGGLVQPGGSLRLSCATSGFTFTSYWMHWVRQAPGKGLEWVAEINPTNG RTNYIEKFKSRITLSVDKSKSTVYLQMNSLRAEDTAVYYCARGTRAYHYWGQGTLV TVSS (SEQ ID NO: 755).

34. The RNAi agent of any one of claims 28-33, wherein the Fab further comprises a light constant chain 1 (CL).

35. The RNAi agent of claim 34 wherein the light constant chain 1 (CL) sequence is: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 717).

36. The RNAi agent of any one of claims 28-35, wherein the Fab further comprises a heavy constant chain 1 (CH).

37. The RNAi agent of claim 36 wherein the heavy constant chain 1 (CH) sequence is: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH (SEQ ID NO: 719).

38. The RNAi agent of any one of claims 28-37, wherein the antibody fragment (Fab) binds TfRl with an affinity of at least 1 nM KD.

39. A conjugate comprising the RNAi agent of any one of claims 1-21 conjugated to an antibody fragment (Fab) that specifically binds to one or more epitopes on a transferrin receptor (TfRl).

40. The conjugate of claim 39, wherein the Fab comprises (i) 6 complementary determining regions (CDRs), (ii) 3 CDRs on the variable light chain (VL), or (iii) 3 CDRs on the variable heavy chain (VH).

41. The conjugate of claim 40, wherein the variable light chain has a VL CDR1 sequence selected from the group consisting of: RASDGLYSNLA (SEQ ID NO: 721), RASDNLYRNLA (SEQ ID NO: 722). and RASDKLYSNLA (SEQ ID NO: 723); a VL CDR2 sequence selected from the group consisting of: DATLLAS (SEQ ID NO: 724), DARNLAS (SEQ ID NO: 725), DAFNLAS (SEQ ID NO: 726), DATRLAS (SEQ ID NO: 727), DATKLAS (SEQ ID NO: 728), and DAKNLAS (SEQ ID NO: 729); and / or a VL CDR 3 sequence of QHFWGTPLT (SEQ ID NO: 730).

42. The conjugate of claim 40 or 41, wherein the variable light chain comprises the sequence:DIQLTQSPSSLSASVGDRVTITCRASDKLYSNLAWYQQKPGKAPKLLIYDATLLASGV PSRFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGQGTKVEIK (SEQ ID NO: 747).

43. The conjugate of any one of claims 40-42. wherein the variable heavy chain has a VH CDR1 sequence selected from the group consisting of: GYTFNSYWMH (SEQ ID NO: 731), GYTFKSYWMH (SEQ ID NO: 732), GFTFTSYWMH (SEQ ID NO: 733), GYTFTSYWVH (SEQ ID NO: 734), and GYTFTSYWMH (SEQ ID NO: 735), a VH CDR2 sequence selected from the group consisting of: EINPTNGRVNYIEKFKS (SEQ ID NO: 736), EINPTNGRFNYIEKFKS (SEQ ID NO: 737), EINPTNGRTNYIEKFKS (SEQ ID NO: 738), and EINPTNGRSNYIEKFKS (SEQ ID NO: 739); and / or a VH CDR3 sequence of: GTRAYHY (SEQ ID NO: 740).

44. The conjugate of any one of claims 40-43, wherein the variable heavy chain comprises the sequence:EVQLVESGGGLVQPGGSLRLSCATSGFTFTSYWMHWVRQAPGKGLEWVAEINPTNG RTNYIEKFKSRITLSVDKSKSTVYLQMNSLRAEDTAVYYCARGTRAYHYWGQGTLV TVSS (SEQ ID NO: 755)45. The conjugate of any one of claims 39-44 wherein the Fab further comprises a light constant chain 1 (CL).

46. The conjugate of claim 45 wherein the light constant chain 1 (CL) sequence is: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 717).

47. The conjugate of any one of claims 39-46, wherein the Fab further comprises a heavy constant chain 1 (CH).

48. The conjugate of claim 47 wherein the heavy constant chain 1 (CH) sequence is: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH (SEQ ID NO: 719).

49. The conjugate of any one of claims 39-48, wherein the antibody fragment (Fab) binds TfRl with an affinity of at least 1 nM KD.

50. The conjugate of claim any one of claims 39-49. wherein the RNAi agent is conjugated to the Fab using a covalent or non-covalent bond, ionic bond, hydrogen bond, hydrophobic interaction, peptide, polymer, or a nucleic acid binding protein.

51. The conjugate of any one of claims 39-50, wherein the RNAi agent is conjugated to the Fab through a linker comprising a structure selected from the group consisting of:wherein5A represents a point of attachment to the Fab, and * R represents a point of attachment to the RNAi agent portion of the conjugate.

52. The RNAi agent of any one of claims 1-21, wherein the RNAi agent is linked to a targeting ligand.

53. The RNAi agent of claim 52, wherein the targeting ligand is linked to the sense strand.

54. The RNAi agent of claim 53, wherein the targeting ligand is linked to the 5?terminal end of the sense strand.

55. The RNAi agent of any of claims 52-54, wherein the targeting ligand has affinity for a skeletal muscle cell and / or a cell receptor expressed on a skeletal muscle cell.

56. The RNAi agent of any of claims 52-55, wherein the targeting ligand has affinity for an integrin alpha-v-beta 6 (avP6) receptor.

57. The RNAi agent of any of claims 52-56. wherein the targeting ligand is:or a pharmaceutically acceptable salt thereof, wherein ? indicates the point of connection to the RNAi agent.

58. The RNAi agent of any one of claims 1-21 or 52-57, wherein the RNAi agent is further linked to a pharmacokinetic / pharmacodynamic (PK / PD) modulator.

59. The RNAi agent of claim 58, wherein the PK / PD modulator is linked to the sense strand.

60. The RNAi agent of claim 59. wherein the PK / PD modulator is linked to the 3’ terminal end of the sense strand.

61. The RNAi agent of any one of claims 58-60, wherein the PK / PD modulator is selected from the group consisting of:wherein Rz comprises the RNAi agent.

62. A composition comprising the RNAi agent of any one of claims 1-38 or 52-61, or the conjugate of any one of claims 39-51, wherein the composition further comprises a pharmaceutically acceptable excipient.

63. The composition of claim 62, further comprising a second RNAi agent capable of inhibiting the expression of Androgen Receptor gene expression.

64. The composition of any one of claims 62-63, further comprising one or more additional therapeutics.

65. The composition of any of claims 62-64, wherein the RNAi agent is a sodium salt.

66. The composition of any of claims 62-65, wherein the pharmaceutically acceptable excipient is water for injection.

67. The composition of any of claims 62-65, wherein the pharmaceutically acceptable excipient is a buffered saline solution.

68. A method for inhibiting expression of an AR gene in a cell, the method comprising introducing into a cell an effective amount of the RNAi agent of any one of claims 1-38 or 52-61, the conjugate of any one of claims 39-51, or the composition of any one of claims 62- 67.

69. The method of claim 68, wherein the cell is within a subject.

70. The method of claim 69, wherein the subject is a human subject.71 . The method of any one of claims 68-70, wherein following the administration of the RNAi agent the Androgen Receptor (AR) gene expression is inhibited by at least about 30%.

72. A method of treating one or more symptoms or diseases associated with enhanced or elevated membrane AR activity levels, the method comprising administering to a human subject in need thereof a therapeutically effective amount of the composition of any one of claims 62-67.

73. The method of claim 72. wherein the disease is a neurodegenerative disease.

74. The method of claim 73, wherein the neurodegenerative disease is spinal and bulbar muscular atrophy (SBMA).

75. The method of any one of claims 68-74, wherein the RNAi agent is administered at a deposited dose of about 0.01 mg / kg to about 5.0 mg / kg of body weight of the subject.

76. The method of any one of claims 68-75, wherein the RNAi agent is administered at a deposited dose of about 0.03 mg / kg to about 2.0 mg / kg of body weight of the subject.

77. The method of any of claims 68-76, wherein the RNAi agent is administered in two or more doses.

78. Use of the RNAi agent of any one of claims 1-38 or 52-61, or the conjugate of any one of claims 39-51, for the treatment of a disease, disorder, or symptom that is mediated at least in part by mutant AR activity and / or AR gene expression.

79. Use of the composition according to any one of claims 62-67, for the treatment of a disease, disorder, or symptom that is mediated at least in part by Androgen Receptor (AR) activity and / or Androgen Receptor (AR) gene expression.

80. Use of the composition according to any one of claims 62-67, for the manufacture of a medicament for treatment of a disease, disorder, or symptom that is mediated at least in part by Androgen Receptor (AR) and / or Androgen Receptor (AR) gene expression.

81. The use of any one of claims 78-80, wherein the disease is a neurodegenerative disease.

82. A method of making an RNAi agent of any one of claims 1-38 or 52-61, comprising annealing a sense strand and an antisense strand to form a double-stranded ribonucleic acid molecule.

83. The method of claim 82. wherein the sense strand comprises a lipid moiety.

84. The method of claim 82, wherein the sense strand comprises an antigen binding moiety.

85. The method of claim 82, wherein the sense strand comprises a targeting ligand.

86. The method of claim 85, wherein the sense strand further comprises a PK / PD modulator.

87. The method of claim 83, comprising conjugating a lipid moiety to the sense strand.

88. The method of claim 84. comprising conjugating an antigen binding moiety to the sense strand.

89. The method of claim 85, comprising conjugating a targeting ligand to the sense strand.

90. The method of claim 86, comprising conjugating a PK / PD modulator to the sense strand.