SARM1 RNA interference reagents

Abstract

Provided herein are SARM1 RNAi agents and compositions comprising a SARM1 RNAi agent. Also provided herein are methods of using the SARM1 RNAi agent or compositions comprising the SARM1 RNAi agent to reduce SARM1 expression, reduce axial degeneration, and / or treat a SARM1 mediated neurological disease in a subject.

Description

[0001] Sequence Listing

[0002] This application is filed with a sequence listing in ST.26 XML format. The sequence listing is provided as a file named "30243", which was created on January 13, 2023 and is 361 kilobytes in size. The sequence listing information in ST.26 XML format is incorporated herein by reference in its entirety. Background Art

[0003] Axonal degeneration is a pathological feature of many neurological diseases, including peripheral neuropathy, traumatic brain injury and neurodegenerative diseases (Gerdts et al., Science, 2015, 348: 453-457). Damaged or unhealthy axons are eliminated by an intrinsic self-destructive program called Wallerian degeneration, which is an active process of retrograde degeneration of the distal end of the axon while maintaining the integrity of the proximal axon segment and cell body (Gerdts, J. et al., Neuron, 2016, 89, 449-460; Whitmore, A. et al., Cell Death Differ., 2003, 10, 260-261).

[0004] SARM1 (protein 1 containing sterile α and TIR motifs) is an NAD+ hydrolase that plays an important role in axonal degeneration. It is reported that knocking down or eliminating the expression of SARM1 can produce a lasting protective effect on sensory neurons against axonal degeneration caused by injury (Gerdts et al., J Neurosci, 2013, 33, 13569-13580). WO2021 / 108602 describes SARM1 antisense oligonucleotides.

[0005] RNA interference (RNAi) is a highly conserved regulatory mechanism in which RNA molecules engage in sequence-specific inhibition of gene expression by double-stranded RNA molecules (dsRNA) (Fire et al., Nature 391: 806-811, 1998).

[0006] There remains a need for therapeutic agents capable of inhibiting or modulating SARM1 expression to treat SARM1-mediated neurological diseases, for example by utilizing RNAi. Summary of the invention

[0007] Provided herein are SARM1 RNAi agents and compositions comprising SARM1 RNAi agents. Also provided herein are methods of using the SARM1 RNAi agents or compositions comprising SARM1 RNAi agents to reduce SARM1 expression, reduce axonal degeneration, and / or treat SARM1-mediated neurological diseases in a subject.

[0008] In one aspect, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the group consisting of:

[0009] (a) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 2;

[0010] (b) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 3, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 4;

[0011] (c) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:5, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:6;

[0012] (d) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:7, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:8;

[0013] (e) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 9, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 10; and

[0014] (f) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 11, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 12,

[0015] wherein optionally one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally one or more internucleotide linkages of the sense strand and the antisense strand are modified internucleotide linkages.

[0016] In some embodiments, the sense strand and antisense strand of the SARM1 RNAi agents described herein comprise a pair of nucleic acid sequences selected from the group consisting of:

[0017] (a) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 2;

[0018] (b) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 3, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 4;

[0019] (c) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 5, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 6;

[0020] (d) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 7, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 8;

[0021] (e) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 9, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 10; and

[0022] (f) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 11, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 12,

[0023] wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0024] In some embodiments, the sense strand and antisense strand of the SARM1 RNAi agent described herein have a pair of nucleic acid sequences selected from the following:

[0025] (a) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 2;

[0026] (b) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 3, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 4;

[0027] (c) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 5, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 6;

[0028] (d) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 7, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 8;

[0029] (e) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 9, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 10; and

[0030] (f) the sense strand has (eg, consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:11, and the antisense strand has (eg, consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:12.

[0031] The SARM1 RNAi agents described herein may include modifications. Modifications may be made to one or more nucleotides of the sense strand and / or antisense strand or to internucleotide bonds. In some embodiments, one or more nucleotides of the sense strand are modified nucleotides. In some embodiments, each nucleotide of the sense strand is a modified nucleotide. In some embodiments, one or more nucleotides of the antisense strand are modified nucleotides. In some embodiments, each nucleotide of the antisense strand is a modified nucleotide. In some embodiments, the modified nucleotides are 2′-fluoro modified nucleotides, 2′-O-methyl modified nucleotides, or 2′-O-alkyl modified nucleotides, such as 2′-OC 12-16Alkyl modified nucleotides. In some embodiments, the sense strand has four 2'-fluoro modified nucleotides at the 7th, 9th, 10th and 11th positions from the 5' end of the sense strand. In some embodiments, the nucleotides at positions other than the 7th, 9th, 10th and 11th on the sense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has four 2'-fluoro modified nucleotides at the 2nd, 6th, 14th and 16th positions from the 5' end of the antisense strand. In some embodiments, the nucleotides at positions other than the 2nd, 6th, 14th and 16th positions on the antisense strand are 2'-O-methyl modified nucleotides. In some embodiments, the sense strand has three 2'-fluoro modified nucleotides at the 9th, 10th and 11th positions from the 5' end of the sense strand. In some embodiments, the nucleotides at positions other than the 9th, 10th and 11th positions on the sense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has five 2'-fluoro modified nucleotides at positions 2, 5, 7, 14, and 16 from the 5' end of the antisense strand. In some embodiments, the nucleotides at positions other than positions 2, 5, 7, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has five 2'-fluoro modified nucleotides at positions 2, 5, 8, 14, and 16 from the 5' end of the antisense strand. In some embodiments, the nucleotides at positions other than positions 2, 5, 8, 14, and 16 of the antisense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has five 2'-fluoro modified nucleotides at positions 2, 3, 7, 14, and 16 from the 5' end of the antisense strand. In some embodiments, the nucleotides at positions other than positions 2, 3, 7, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has two 2'-fluoro modified nucleotides at positions 2 and 14 from the 5' end of the antisense strand. In some embodiments, nucleotides at positions other than positions 2 and 14 on the antisense strand are 2'-O-methyl modified nucleotides.

[0032] In some embodiments, the first nucleotide at the 5' end of the antisense strand is a modified nucleotide having a phosphate analog, such as 5'-vinylphosphonate. In some embodiments, the first nucleotide at the 5' end of the antisense strand has a 5' phosphate. In some embodiments, the sense strand comprises an abasic portion or a reversed abasic portion.

[0033] In some embodiments, the sense strand and the antisense strand have one or more modified internucleotide bonds, such as thiophosphate bonds. In some embodiments, the sense strand has four or five thiophosphate bonds. In some embodiments, the antisense strand has four or five thiophosphate bonds. In some embodiments, the sense strand has four thiophosphate bonds and the antisense strand has four thiophosphate bonds.

[0034] In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the group consisting of:

[0035] (a) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 14;

[0036] (b) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 15, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 16;

[0037] (c) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 17, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 18;

[0038] (d) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 19, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 20;

[0039] (e) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 21, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 22;

[0040] (f) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 23, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 24;

[0041] (g) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 15, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 27;

[0042] (h) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 29;

[0043] (i) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 30;

[0044] (j) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 31;

[0045] (k) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 32;

[0046] (1) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 33;

[0047] (m) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 35;

[0048] (n) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 36;

[0049] (o) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 37;

[0050] (p) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 38;

[0051] (q) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 21, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 39;

[0052] (r) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 41;

[0053] (s) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 42;

[0054] (t) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 43;

[0055] (u) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 44;

[0056] (v) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 17, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 45;

[0057] (w) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 19, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 46; and

[0058] (x) The sense strand comprises the first nucleic acid sequence of SEQ ID NO:23, and the antisense strand comprises the second nucleic acid sequence of SEQ ID NO:47.

[0059] In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, wherein the sense strand and the antisense strand have a pair of nucleic acid sequences selected from the following:

[0060] (a) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 14;

[0061] (b) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 16;

[0062] (c) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 18;

[0063] (d) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 20;

[0064] (e) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 22;

[0065] (f) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 24;

[0066] (g) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 27;

[0067] (h) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 29;

[0068] (i) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 30;

[0069] (j) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:31;

[0070] (k) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:32;

[0071] (l) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 33;

[0072] (m) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 35;

[0073] (n) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:36;

[0074] (o) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:37;

[0075] (p) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:38;

[0076] (q) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:21, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:39;

[0077] (r) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:41;

[0078] (s) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:42;

[0079] (t) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:43;

[0080] (u) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:44;

[0081] (v) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 45;

[0082] (w) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 46; and

[0083] (x) the sense strand has (eg, consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:23, and the antisense strand has (eg, consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:47.

[0084] In some embodiments, the sense strand of the SARM1 RNAi agent has a delivery portion. In some embodiments, the sense strand of the SARM1 RNAi agent has a delivery portion conjugated to the 3' end of the sense strand. In some embodiments, the sense strand of the SARM1 RNAi agent has a delivery portion conjugated to the 5' end of the sense strand. In some embodiments, the sense strand of the SARM1 RNAi agent has a delivery portion conjugated to both the 5' end and the 3' end of the sense strand. In some embodiments, the sense strand of the SARM1 RNAi agent has a delivery portion conjugated to the nucleotides of the sense strand. In some embodiments, the delivery portion is alpha-tocopherol, palmitic acid, or another portion in Table 4. In some embodiments, the delivery portion is conjugated to the 5' or 3' end of the sense strand through a linker (e.g., a linker of Table 5). In some embodiments, the SARM1 RNAi agent is not a lipid conjugate. In some embodiments, the SARM1 RNAi agent has no delivery portion.

[0085] In a further aspect, provided herein is a SARM1 RNAi agent of formula (I): RLD, wherein R is a double-stranded RNA (dsRNA) having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex; wherein D is a delivery means (e.g., a delivery moiety) for delivering the dsRNA into a cell; and wherein L is a linking means (e.g., a linker) for linking the dsRNA to the delivery means, or alternatively is absent, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the following:

[0086] (a) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 2;

[0087] (b) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 3, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 4;

[0088] (c) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 5, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 6;

[0089] (d) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 7, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 8;

[0090] (e) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 9, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 10; and

[0091] (f) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 11, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 12,

[0092] wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0093] In some embodiments, provided herein is a SARM1 RNAi agent of formula (I): RLD, wherein R is a dsRNA having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex; wherein D is a delivery means (e.g., a delivery moiety) for delivering the dsRNA into a cell; and wherein L is a connection means (e.g., a linker) for connecting the dsRNA to the delivery means, or alternatively is absent, wherein the sense strand and the antisense strand have a pair of nucleic acid sequences selected from the following:

[0094] (a) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 2;

[0095] (b) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 3, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 4;

[0096] (c) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 5, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 6;

[0097] (d) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 7, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 8;

[0098] (e) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 9, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 10; and

[0099] (f) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 11, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 12,

[0100] wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0101] In some embodiments, provided herein is a SARM1 RNAi agent of formula (I): RLD, wherein R is a dsRNA having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex; wherein D is a delivery means (e.g., a delivery moiety) for delivering the dsRNA into a cell; and wherein L is a connection means (e.g., a linker) for connecting the dsRNA to the delivery means, or alternatively is absent, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the following:

[0102] (a) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 14;

[0103] (b) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 15, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 16;

[0104] (c) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 17, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 18;

[0105] (d) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 19, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 20;

[0106] (e) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 21, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 22;

[0107] (f) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 23, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 24;

[0108] (g) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 15, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 27;

[0109] (h) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 29;

[0110] (i) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 30;

[0111] (j) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 31;

[0112] (k) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 32;

[0113] (1) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 33;

[0114] (m) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 35;

[0115] (n) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 36;

[0116] (o) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 37;

[0117] (p) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 38;

[0118] (q) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 21, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 39;

[0119] (r) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 41;

[0120] (s) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 42;

[0121] (t) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 43;

[0122] (u) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 44;

[0123] (v) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 17, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 45;

[0124] (w) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 19, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 46; and

[0125] (x) The sense strand comprises the first nucleic acid sequence of SEQ ID NO:23, and the antisense strand comprises the second nucleic acid sequence of SEQ ID NO:47.

[0126] In some embodiments, provided herein is a SARM1 RNAi agent of formula (I): RLD, wherein R is a dsRNA having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex; wherein D is a delivery means (e.g., a delivery moiety) for delivering the dsRNA into a cell; and wherein L is a connection means (e.g., a linker) for connecting the dsRNA to the delivery means, or alternatively is absent, wherein the sense strand and the antisense strand have a pair of nucleic acid sequences selected from the following:

[0127] (a) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 14;

[0128] (b) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 16;

[0129] (c) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 18;

[0130] (d) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 20;

[0131] (e) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 22;

[0132] (f) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 24;

[0133] (e) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 27;

[0134] (f) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 29;

[0135] (g) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:30;

[0136] (h) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:31;

[0137] (i) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 32;

[0138] (j) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 33;

[0139] (k) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:35;

[0140] (1) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 36;

[0141] (m) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 37;

[0142] (n) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:38;

[0143] (o) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:21, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:39;

[0144] (p) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:41;

[0145] (q) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:42;

[0146] (r) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:43;

[0147] (s) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:44;

[0148] (t) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 45;

[0149] (u) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 46; and

[0150] (v) the sense strand has (eg, consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:23, and the antisense strand has (eg, consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:47.

[0151] In another aspect, provided herein is a pharmaceutical composition comprising a SARM1 RNAi agent as described herein and a pharmaceutically acceptable carrier. Also provided herein is a pharmaceutical composition comprising a means for reducing SARM1 expression in a cell and a pharmaceutically acceptable carrier.

[0152] In another aspect, provided herein are methods of reducing SARM1 expression in a patient in need thereof, and such methods comprise administering to the patient an effective amount of a SARM1 RNAi agent or pharmaceutical composition described herein.

[0153] In another aspect, provided herein are methods of reducing axonal degeneration in a patient in need thereof, and such methods comprise administering to the patient an effective amount of a SARM1 RNAi agent or pharmaceutical composition described herein.

[0154] In another aspect, provided herein are methods of treating a SARM1-mediated neurological disease in a patient in need thereof, and such methods comprise administering to the patient an effective amount of a SARM1 RNAi agent or pharmaceutical composition described herein.

[0155] Also provided herein are methods of reducing SARM1 expression in cells (eg, neurons), and such methods may include introducing a SARM1 RNAi agent described herein into the cells; and incubating the cells for a time sufficient to degrade SARM1 mRNA, thereby reducing SARM1 expression in the cells.

[0156] In another aspect, provided herein is a SARM1 RNAi agent for reducing SARM1 expression or a pharmaceutical composition comprising a SARM1 RNAi agent. In another aspect, provided herein is a SARM1 RNAi agent for reducing axonal degeneration or a pharmaceutical composition comprising a SARM1 RNAi agent. Also provided herein is a SARM1 RNAi agent for use in therapy or a pharmaceutical composition comprising a SARM1 RNAi agent. Also provided herein is a SARM1 RNAi agent for treating a SARM1-mediated nervous system disease or a pharmaceutical composition comprising a SARM1 RNAi agent. Also provided herein is the use of a SARM1 RNAi agent in the preparation of a medicament for reducing axonal degeneration. Also provided herein is the use of a SARM1 RNAi agent in the preparation of a medicament for treating a SARM1-mediated nervous system disease.

[0157] Detailed Description

[0158] Provided herein are SARM1 RNAi agents and compositions comprising SARM1 RNAi agents. Also provided herein are methods of using the SARM1 RNAi agents or compositions comprising SARM1 RNAi agents to reduce SARM1 expression, reduce axonal degeneration, and / or treat SARM1-mediated neurological diseases in a subject.

[0159] In some embodiments, provided herein is a SARM1 RNAi agent with a sense strand and an antisense strand, and the sense strand and the antisense strand form a duplex. The antisense strand is complementary to the region of SARM1 mRNA. In a further embodiment, the length of the sense strand and the antisense strand is 15-30 nucleotides, for example, 20-25 nucleotides in length. In some embodiments, provided herein is a SARM1 RNAi agent with a sense strand of 21 nucleotides and an antisense strand of 23 nucleotides. In some embodiments, the sense strand and the antisense strand of the SARM1 RNAi agent may have an overhang (i.e., a 5' overhang or a 3' overhang) at the 5' end or the 3' end. For example, the sense strand and the antisense strand may have a 5' or 3' overhang of 1 to 5 nucleotides or 1 to 3 nucleotides. In some embodiments, the antisense strand comprises a 3' overhang of two nucleotides. In some embodiments, the sense strand and the antisense strand sequences of the SARM1 RNAi agent are provided in Table 1.

[0160] Provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the group consisting of:

[0161] (a) the sense strand comprises a first nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 2;

[0162] (b) the sense strand comprises a first nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 3, and the antisense strand comprises a second nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 4;

[0163] (c) the sense strand comprises a first nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:5, and the antisense strand comprises a second nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:6;

[0164] (d) the sense strand comprises a first nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:7, and the antisense strand comprises a second nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:8;

[0165] (e) the sense strand comprises a first nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 9, and the antisense strand comprises a second nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 10; and

[0166] (f) the sense strand comprises a first nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 11, and the antisense strand comprises a second nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 12,

[0167] wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0168] Provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the group consisting of:

[0169] (a) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 2;

[0170] (b) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 3, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 4;

[0171] (c) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:5, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:6;

[0172] (d) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:7, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:8;

[0173] (e) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 9, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 10; and

[0174] (f) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 11, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 12,

[0175] wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0176] In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 2, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 3, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 4, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 5, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 6, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 7, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 8, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 9, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 10, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 11, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 12, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0177] Table 1: Unmodified SARM1 RNAi reagent nucleic acid sequences

[0178]

[0179] *Nucleotides 2-23 of the antisense strand are complementary to human SARM1 mRNA.

[0180] In some embodiments, the sense strand and antisense strand of the SARM1 RNAi agents described herein comprise a pair of nucleic acid sequences selected from the group consisting of:

[0181] (a) the sense strand comprises a first nucleic acid sequence SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence SEQ ID NO: 2;

[0182] (b) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 3, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 4;

[0183] (c) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 5, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 6;

[0184] (d) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 7, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 8;

[0185] (e) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 9, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 10; and

[0186] (f) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 11, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 12,

[0187] wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0188] In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 2, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 3, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 4, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 5, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 6, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 7, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 8, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 9, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 10, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 11, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 12, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0189] In some embodiments, the sense strand and antisense strand of the SARM1 RNAi agent described herein have a pair of nucleic acid sequences selected from the following:

[0190] (a) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 2;

[0191] (b) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 3, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 4;

[0192] (c) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 5, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 6;

[0193] (d) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 7, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 8;

[0194] (e) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 9, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 10; and

[0195] (f) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 11, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 12,

[0196] wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0197] In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 2, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 3, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 4, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 5, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 6, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 7, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 8, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 9, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 10, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 11, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 12, wherein one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0198] The SARM1 RNAi agents described herein may include modifications. Modifications may be made to one or more nucleotides of the sense strand and / or antisense strand or to the internucleotide bond, which is the bond between two nucleotides in the sense strand or antisense strand. For example, some 2'-modifications of ribose or deoxyribose can increase the stability and half-life of RNA or DNA. Such 2'-modifications can be 2'-fluoro, 2'-O-methyl (i.e., 2'-methoxy), 2'-O-alkyl, or 2'-O-methoxyethyl (2'-O-MOE).

[0199] In some embodiments, one or more nucleotides of the sense strand and / or antisense strand are independently modified nucleotides, which means that the sense strand and the antisense strand can have different modified nucleotides. In some embodiments, one or more nucleotides of the sense strand are modified nucleotides. In some embodiments, each nucleotide of the sense strand is a modified nucleotide. In some embodiments, one or more nucleotides of the antisense strand are modified nucleotides. In some embodiments, each nucleotide of the antisense strand is a modified nucleotide. In some embodiments, the modified nucleotides are 2′-fluoro modified nucleotides, 2′-O-methyl modified nucleotides, or 2′-O-alkyl modified nucleotides, such as 2′-OC 12-16 Alkyl modified nucleotides. In some embodiments, each nucleotide of the sense strand and the antisense strand is independently a modified nucleotide, such as a 2′-fluoro modified nucleotide, a 2′-O-methyl modified nucleotide, or a 2′-O-alkyl modified nucleotide, such as a 2′-OC 12-16 Alkyl-modified nucleotides.

[0200] In some embodiments, the sense strand has four 2'-fluoro modified nucleotides, for example, at positions 7, 9, 10, and 11 from the 5' end of the sense strand. In some embodiments, the nucleotides at positions other than positions 7, 9, 10, and 11 on the sense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has four 2'-fluoro modified nucleotides, for example, at positions 2, 6, 14, and 16 from the 5' end of the antisense strand. In some embodiments, the nucleotides at positions other than positions 2, 6, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides. In some embodiments, the sense strand has three 2'-fluoro modified nucleotides at positions 9, 10, and 11 from the 5' end of the sense strand. In some embodiments, the nucleotides at positions other than positions 9, 10, and 11 on the sense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has five 2'-fluoro modified nucleotides at positions 2, 5, 7, 14, and 16 from the 5' end of the antisense strand. In some embodiments, the nucleotides at positions other than positions 2, 5, 7, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has five 2'-fluoro modified nucleotides at positions 2, 5, 8, 14, and 16 from the 5' end of the antisense strand. In some embodiments, the nucleotides at positions other than positions 2, 5, 8, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has five 2'-fluoro modified nucleotides at positions 2, 3, 7, 14, and 16 from the 5' end of the antisense strand. In some embodiments, the nucleotides at positions other than positions 2, 3, 7, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides. In some embodiments, the antisense strand has two 2'-fluoro modified nucleotides at positions 2 and 14 from the 5' end of the antisense strand. In some embodiments, nucleotides at positions other than positions 2 and 14 on the antisense strand are 2'-O-methyl modified nucleotides.

[0201] In some embodiments, the modified nucleotides are 2′-O-alkyl modified nucleotides, such as 2′-OC 12-16 Alkyl modified nucleotides, which can be used as delivery moieties. In some embodiments, the 2′-O-alkyl modified nucleotide is 2′-O-hexadecyluridine, 2′-O-hexadecylcytidine, 2′-O-hexadecylguanine, or 2′-O-hexadecyladenosine. In some embodiments, 2′-O-hexadecyluridine, 2′-O-hexadecylcytidine, 2′-O-hexadecylguanine, or 2′-O-hexadecyladenosine is a modified nucleotide in the sense strand.

[0202] In some embodiments, the first nucleotide at the 5' end of the antisense strand is a modified nucleotide with a phosphate analog, such as 5'-vinylphosphonate (5'-VP). In some embodiments, the first nucleotide at the 5' end of the antisense strand has a 5' phosphate.

[0203] In some embodiments, the sense strand comprises an abasic portion or an inverted abasic portion, such as those shown in Table 3.

[0204] In some embodiments, the sense strand and the antisense strand have one or more modified internucleotide bonds. In some embodiments, the modified internucleotide bond is a phosphorothioate bond. In some embodiments, the sense strand has four or five phosphorothioate bonds. In some embodiments, the antisense strand has four or five phosphorothioate bonds. In some embodiments, the sense strand and the antisense strand each have four or five phosphorothioate bonds. In some embodiments, in some embodiments, the sense strand has four phosphorothioate bonds and the antisense strand has four phosphorothioate bonds.

[0205] In a further aspect, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the group consisting of:

[0206] (a) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 14;

[0207] (b) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 15, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 16;

[0208] (c) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 17, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 18;

[0209] (d) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 19, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 20;

[0210] (e) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 21, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 22;

[0211] (f) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 23, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 24;

[0212] (g) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 15, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 27;

[0213] (h) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 29;

[0214] (i) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 30;

[0215] (j) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 31;

[0216] (k) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 32;

[0217] (1) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 33;

[0218] (m) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 35;

[0219] (n) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 36;

[0220] (o) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 37;

[0221] (p) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 38;

[0222] (q) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 21, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 39;

[0223] (r) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 41;

[0224] (s) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 42;

[0225] (t) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 43;

[0226] (u) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 44;

[0227] (v) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 17, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 45;

[0228] (w) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 19, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 46; and

[0229] (x) The sense strand comprises the first nucleic acid sequence of SEQ ID NO:23, and the antisense strand comprises the second nucleic acid sequence of SEQ ID NO:47.

[0230] In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 14. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 16. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 18. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 20. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 22. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 24. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 27. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 29.In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 30. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 31. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 32. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 33. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 35. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 36. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 37. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 38.In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 39. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 41. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 42. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 43. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 44. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 45. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 46. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 47.

[0231] In a further aspect, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, wherein the sense strand and the antisense strand have a pair of nucleic acid sequences selected from the group consisting of:

[0232] (a) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 14;

[0233] (b) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 16;

[0234] (c) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 18;

[0235] (d) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 20;

[0236] (e) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 22;

[0237] (f) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 24;

[0238] (g) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 27;

[0239] (h) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 29;

[0240] (i) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 30;

[0241] (j) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:31;

[0242] (k) the sense strand comprises (e.g., consists of or consists essentially of) a first nucleic acid

[0243] (1) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 32;

[0244] (m) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 33;

[0245] (n) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 35;

[0246] (o) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:36;

[0247] (p) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:37;

[0248] (q) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:38;

[0249] (r) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:21, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:39;

[0250] (s) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:41;

[0251] (t) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:42;

[0252] (u) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:43;

[0253] (v) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:44;

[0254] (v) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 45;

[0255] (w) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 46; and

[0256] (x) the sense strand has (eg, consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:23, and the antisense strand has (eg, consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:47.

[0257] In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 14. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 16. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 18. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 20. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 22. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 24. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 27. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 29.In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 30. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 31. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 32. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 33. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 35. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 36. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 37. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 38.In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 39. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 41. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 42. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 43. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 44. In some embodiments, provided herein are SARM1 RNAi agents having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 45. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 46. In some embodiments, provided herein is a SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, and wherein the sense strand has a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 47.

[0258] Table 2: Modified SARM1 RNAi Reagents

[0259]

[0260]

[0261]

[0262] Abbreviations—“m” denotes 2′-OMe; “f” denotes 2′-fluoro; “*” denotes a phosphorothioate linkage; “VP” denotes 5′-vinylphosphonate; “S” denotes the sense strand; “AS” denotes the antisense strand; unless otherwise indicated, the 5′ position of AS may include either a 5′-phosphate or a 5′-vinylphosphonate.

[0263] Table 3. Abasic part or inverted abasic (iAb) part

[0264]

[0265] "5'" and "3'" indicate the 5' to 3' direction of the sequence.

[0266] In some embodiments, the sense strand of a SARM1 RNAi agent has a delivery moiety. In some embodiments, the sense strand of a SARM1 RNAi agent has a delivery moiety conjugated to the 3' end of the sense strand. In some embodiments, the sense strand of a SARM1 RNAi agent has a delivery moiety conjugated to the 5' end of the sense strand. In some embodiments, the sense strand of a SARM1 RNAi agent has a delivery moiety conjugated to both the 5' end and the 3' end of the sense strand. In some embodiments, the sense strand of a SARM1 RNAi agent has a delivery moiety conjugated to the nucleotides of the sense strand. The delivery moiety can facilitate entry of the RNAi agent into cells. In some embodiments, the delivery moiety is alpha-tocopherol, palmitic acid, or another moiety in Table 4. In some embodiments, the delivery moiety is a known delivery moiety for delivering RNAi agents into cells. Placing the delivery moiety on an RNAi agent requires overcoming the potential inefficient loading of AGO2 (Argonaute-2), or other obstacles to the activity of the RNA-induced silencing complex (RISC) complex. In some embodiments, the SARM1 RNAi agent is not a lipid conjugate. In some embodiments, the SARM1 RNAi agent is devoid of a delivery moiety.

[0267] In some embodiments, the delivery moiety is conjugated to the 5' or 3' end of the sense strand via a linker. In some embodiments, the linker is selected from linker 1, linker 2, linker 3, or linker 4 of Table 5. Other suitable linkers are known in the art. Exemplary linker-delivery moiety pairs are shown in Table 6. In some embodiments, the SARM1 RNAi agent has a linker-delivery moiety pair of Table 6.

[0268] In some embodiments, the delivery moiety is conjugated to a nucleotide of the sense strand. In this case, the delivery moiety is a modified nucleotide (e.g., 2'-OC 12-16 In some embodiments, the modified nucleotide is 2'-O-hexadecyluridine, 2'-O-hexadecylcytidine, 2'-O-hexadecylguanine, or 2'-O-hexadecyladenosine (Table 4).

[0269] Table 4. Delivery parts

[0270]

[0271]

[0272]

[0273] Table 5. Connectors

[0274]

[0275] Table 6. Linker Delivery Moiety Pairs (LDP)

[0276]

[0277]

[0278]

[0279] In a further aspect, provided herein is a SARM1 RNAi agent of formula (I): RLD, wherein R is a double-stranded RNA (dsRNA) having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex; wherein D is a delivery means (e.g., a delivery moiety) for delivering the dsRNA into a cell; and wherein L is a linking means (e.g., a linker) for linking the dsRNA to the delivery means, or alternatively is absent, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the following:

[0280] (a) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 2;

[0281] (b) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 3, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 4;

[0282] (c) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 5, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 6;

[0283] (d) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 7, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 8;

[0284] (e) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 9, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 10; and

[0285] (f) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 11, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 12,

[0286] wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0287] In some embodiments, provided herein are SARM1 RNAi agents of formula (I): RLD, wherein R is a double-stranded RNA (dsRNA) having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex; wherein D is a delivery vehicle (e.g., a delivery moiety) for delivering the dsRNA into a cell; and wherein L is a linker (e.g., a linker) for linking the dsRNA to the delivery vehicle, or alternatively is absent, wherein the sense strand and the antisense strand have a pair of nucleic acid sequences selected from the following:

[0288] (a) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 2;

[0289] (b) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 3, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 4;

[0290] (c) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 5, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 6;

[0291] (d) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 7, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 8;

[0292] (e) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 9, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 10; and

[0293] (f) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 11, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 12,

[0294] wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and wherein optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

[0295] In some embodiments, provided herein are SARM1 RNAi agents of formula (I): RLD, wherein R is a double-stranded RNA (dsRNA) having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex; wherein D is a delivery means (e.g., a delivery moiety) for delivering the dsRNA into a cell; and wherein L is a connection means (e.g., a linker) for connecting the dsRNA to the delivery means, or alternatively is absent, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the following:

[0296] (a) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 14;

[0297] (b) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 15, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 16;

[0298] (c) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 17, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 18;

[0299] (d) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 19, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 20;

[0300] (e) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 21, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 22;

[0301] (f) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 23, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 24;

[0302] (g) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 15, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 27;

[0303] (h) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 29;

[0304] (i) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 30;

[0305] (j) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 31;

[0306] (k) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 28, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 32;

[0307] (l) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 33;

[0308] (m) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 35;

[0309] (n) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 36;

[0310] (o) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 37;

[0311] (p) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 34, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 38;

[0312] (q) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 21, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 39;

[0313] (r) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 41;

[0314] (s) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 42;

[0315] (t) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 43;

[0316] (u) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 40, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 44;

[0317] (v) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 17, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 45;

[0318] (w) the sense strand comprises the first nucleic acid sequence SEQ ID NO: 19, and the antisense strand comprises the second nucleic acid sequence SEQ ID NO: 46; and

[0319] (x) The sense strand comprises the first nucleic acid sequence of SEQ ID NO:23, and the antisense strand comprises the second nucleic acid sequence of SEQ ID NO:47.

[0320] In some embodiments, provided herein are SARM1 RNAi agents of formula (I): RLD, wherein R is a double-stranded RNA (dsRNA) having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex; wherein D is a delivery vehicle (e.g., a delivery moiety) for delivering the dsRNA into a cell; and wherein L is a linker (e.g., a linker) for linking the dsRNA to the delivery vehicle, or alternatively is absent, wherein the sense strand and the antisense strand have a pair of nucleic acid sequences selected from the following:

[0321] (a) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 14;

[0322] (b) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 16;

[0323] (c) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 18;

[0324] (d) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 20;

[0325] (e) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 22;

[0326] (f) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 24;

[0327] (g) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 27;

[0328] (h) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 29;

[0329] (i) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 30;

[0330] (j) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:31;

[0331] (k) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:28, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:32;

[0332] (1) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 33;

[0333] (m) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 35;

[0334] (n) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 36;

[0335] (o) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:37;

[0336] (p) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:34, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:38;

[0337] (q) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:21, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:39;

[0338] (r) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:41;

[0339] (s) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:42;

[0340] (t) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:43;

[0341] (u) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:40, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:44;

[0342] (v) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 45;

[0343] (w) the sense strand has (e.g., consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has (e.g., consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO: 46; and

[0344] (x) the sense strand has (eg, consists of or consists essentially of) a first nucleic acid sequence of SEQ ID NO:23, and the antisense strand has (eg, consists of or consists essentially of) a second nucleic acid sequence of SEQ ID NO:47.

[0345] In some embodiments, the delivery means is combined with the 3' end of the sense strand. In some embodiments, the delivery means is conjugated to the 5' end of the sense strand. In some embodiments, the delivery device is conjugated to the nucleotides of the sense strand. In some embodiments, the delivery means is palmitic acid, alpha-tocopherol, or another part in Table 4. In some embodiments, the connection means is selected from the connector 1, connector 2, connector 3 and connector 4 of Table 5.

[0346] The sense and antisense strands of the SARM1 RNAi agent can be synthesized using any nucleic acid polymerization method known in the art, for example, by solid phase synthesis using phosphoramidite chemistry (e.g., Current Protocols in Nucleic Acid Chemistry, Beaucage, SL et al., (Edrs), John Wiley & Sons, Inc., New York, NY, USA), hydrogen phosphite, phosphotriester chemistry, or enzymatic synthesis. An automated commercial synthesizer, such as the MerMade from LGC Biosearch Technologies, can be used. TM 12, or other synthesizers from BioAutomation or Applied Biosystems. Thiolating agents such as phenylacetyl disulfide or DDTT (((dimethylaminomethylene)amino)-3H-1,2,4-dithiazoline-3-thione) can be used to introduce phosphorothioate bonds. It is well known that modified oligonucleotides or conjugated oligonucleotides are synthesized using similar techniques and commercially available modified amides and controlled pore glass (CPG) products.

[0347] Purification methods can be used to exclude unwanted impurities from the final oligonucleotide product. Common purification techniques for single-stranded oligonucleotides include reversed-phase ion-pair high performance liquid chromatography (RP-IP-HPLC), capillary gel electrophoresis (CGE), anion exchange chromatography (AX-HPLC) and size exclusion chromatography (SEC). After purification, oligonucleotides can be analyzed by mass spectrometry and quantitatively analyzed by spectrophotometry at a wavelength of 260nm. Then the sense strand and antisense strand can be annealed to form a duplex.

[0348] In another aspect, provided herein is a pharmaceutical composition comprising a SARM1 RNAi agent described herein and a pharmaceutically acceptable carrier. Also provided herein is a pharmaceutical composition comprising a tool for reducing SARM1 expression in a cell and a pharmaceutically acceptable carrier. Such pharmaceutical compositions may also include one or more pharmaceutically acceptable excipients, diluents or carriers. The pharmaceutical composition can be prepared by methods well known in the art (e.g., Remington: The Science and Practice of Pharmacy, 23rd edition (2020), A. Loyd et al., Academic Press).

[0349] In a further aspect, provided herein is a method of reducing SARM1 expression in a cell (e.g., a neuron), such method may include introducing a SARM1 RNAi agent described herein into a cell; and incubating the cell for a time sufficient to degrade SARM1 mRNA, thereby reducing SARM1 expression in the cell. SARM1 RNAi agents may be introduced into cells (e.g., neurons) using methods known in the art, such as transfection, electroporation, microinjection, or uptake by the cell via natural transport mechanisms.

[0350] In another aspect, provided herein are methods of reducing SARM1 expression in a patient in need thereof, and such methods comprise administering to the patient an effective amount of a SARM1 RNAi agent or pharmaceutical composition described herein.

[0351] In another aspect, provided herein are methods of reducing axonal degeneration in a patient in need thereof, and such methods comprise administering to the patient an effective amount of a SARM1 RNAi agent or pharmaceutical composition described herein.

[0352] In another aspect, provided herein are methods of treating a SARM1-mediated neurological disease in a patient in need thereof, and such methods comprise administering to the patient an effective amount of a SARM1 RNAi agent or pharmaceutical composition described herein. Exemplary SARM1-mediated nervous system diseases include, but are not limited to, amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease), Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), Huntington's disease (HD), Alzheimer's disease, Pick's disease, Gaucher's disease, Hurler syndrome, progressive multifocal leukoencephalopathy, Alexander disease, congenital hypomyelination, encephalomyelitis, acute disseminated encephalomyelitis, central pontine myelinolysis, osmotic hyponatremia, Tay-Sachs disease, motor neuron disease, ataxia, spinal muscular atrophy (SMA), Niemann-Pick disease, acute hemorrhagic leukoencephalitis, trigeminal neuralgia, Bell's palsy, cerebral ischemia, multiple system atrophy, Pelizaeus Merzbacher disease, and schizophrenia. disease), periventricular leukomalacia, hereditary ataxia, noise-induced hearing loss, congenital hearing loss, age-related hearing loss, Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, dementia with Lewy bodies, frontotemporal dementia, tauopathies, synucleinopathies, amyloidosis, diabetic neuropathy, globoid cell leukodystrophy (Krabbe's disease), Bassen-Komzweig syndrome, transverse myelitis, motor neuron disease, spinocerebellar ataxia, preeclampsia, hereditary spastic paraplegia, spastic paraparesis, familial spastic paraparesis, French residency disease, Strumpell-Lorrain disease, nonalcoholic steatohepatitis (NASH), adrenoleukodystrophy, progressive supranuclear palsy (PSP), Friedrich's ataxia, ataxia, spinal cord injury, acute optic neuropathy (AON), hereditary or idiopathic retinal disorders, Leber congenital amaurosis (LCA), Leber hereditary optic neuropathy (LHON), primary open-angle glaucoma (POAG), acute angle-closure glaucoma (AACG), autosomal dominant optic atrophy, retinal ganglion degeneration, retinitis pigmentosa, outer retinal neuropathy, optic neuritis, optic nerve degeneration associated with multiple sclerosis, Kjer's opticneuropathy, ischemic optic neuropathy, chemotherapy-induced peripheral neuropathy, neuromyelitis optica, Charcot Marie Tooth disease, vitamin B12 deficiency, folate (vitamin B9) deficiency, isolated vitamin E deficiency syndrome, nonarteritic anterior ischemic optic neuropathy, ethambutol exposure, cyanide exposure, traumatic brain injury (TBI), spinal cord injury, traumatic axonal injury, or chronic traumatic encephalopathy (CTE). In some embodiments, the SARM1-mediated neurological disease is amyotrophic lateral sclerosis, multiple sclerosis, chemotherapy-induced peripheral neuropathy (CIPN), diabetic peripheral neuropathy (DPN), Tauopathy, or Charcot-Marie-Tooth disease. In some embodiments, the SARM1-mediated neurological disease is amyotrophic lateral sclerosis. ,

[0353] The SARM1 RNAi agent can be administered to the patient intrathecally, intracerebroventricularly, or by intracisternal injection. In some embodiments, the SARM1 RNAi agent is administered to the patient intrathecally, for example by catheter or direct injection into the intrathecal space.

[0354] The RNAi agent dosage regimen can be adjusted to provide the optimal desired response (e.g., therapeutic response). For example, a single bolus can be administered, multiple divided doses can be administered over time, or the dose can be proportionally reduced or increased according to the urgency of the therapeutic situation.

[0355] Dosage values ​​may vary with the type and severity of the condition to be alleviated.It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions.

[0356] In another aspect, provided herein is a SARM1 RNAi agent for reducing SARM1 expression or a pharmaceutical composition comprising a SARM1 RNAi agent. Also provided herein is a SARM1 RNAi agent for reducing axonal degeneration or a pharmaceutical composition comprising a SARM1 RNAi agent. Also provided herein is a SARM1 RNAi agent for use in therapy or a pharmaceutical composition comprising a SARM1 RNAi agent. Also provided herein is a SARM1 RNAi agent for treating a SARM1-mediated nervous system disease or a pharmaceutical composition comprising a SARM1 RNAi agent. Also provided herein is the use of a SARM1 RNAi agent in the preparation of a medicament for reducing axonal degeneration. Also provided herein is the use of a SARM1 RNAi agent in the preparation of a medicament for treating a SARM1-mediated nervous system disease.

[0357] As used herein, the terms "a," "an," "the" and similar terms used in the context of the disclosure (especially in the context of the claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0358] As used herein, the term "alkyl" means a saturated straight or branched chain monovalent hydrocarbon radical containing the indicated number of carbon atoms. For example, "C1-C 20 "Alkyl" means a group having 1 to 20 carbon atoms in a straight chain or branched chain.

[0359] As used herein, "antisense strand" means a single-stranded oligonucleotide complementary to a target sequence region. Likewise, as used herein, "sense strand" means a single-stranded oligonucleotide complementary to an antisense strand region.

[0360] As used herein, "complementarity" means a structural relationship between two nucleotides (e.g., on two relative nucleic acids or on the relative regions of a single nucleic acid strand, such as a hairpin structure), which allows the two nucleotides to form base pairs with each other. For example, the purine nucleotides of a nucleic acid complementary to the pyrimidine nucleotides of the relative nucleic acid can be base-paired together by forming hydrogen bonds with each other. Complementary nucleotides can be base-paired in a Watson-Crick manner or in any other manner that allows the formation of a stable double-stranded chain. Similarly, two nucleic acids can have multiple nucleotide regions that are complementary to each other to form a complementary region, as described herein.

[0361] As used herein, "delivery moiety" refers to a moiety that facilitates the entry of an oligonucleotide or RNAi agent into a cell. The delivery moiety can be a lipid, cholesterol, vitamin E, a carbohydrate, an amino sugar, a polypeptide, or a combination thereof.

[0362] As used herein, "duplex" in reference to a nucleic acid or oligonucleotide means a structure formed by complementary base pairing of two antiparallel nucleotide sequences (i.e., in opposite orientations), whether formed by two covalently separated nucleic acid strands or by a single folded strand (e.g., via a hairpin structure).

[0363] An "effective amount" refers to the amount necessary (for a period of time and for the route of administration) to achieve the desired therapeutic result. The effective amount of an RNAi agent may vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the RNAi agent to elicit a desired response in the individual. An effective amount is also an amount in which the therapeutically beneficial effects of the RNAi agent outweigh any toxic or deleterious effects thereof.

[0364] The term "knockdown" or "expression knockdown" refers to a decrease in mRNA or protein expression of a gene following treatment with an agent (eg, an RNAi agent).

[0365] As used herein, "modified internucleotide bonds" means internucleotide bonds having one or more chemical modifications compared to a reference internucleotide bond having a phosphodiester bond. The modified internucleotide bond can be a non-naturally occurring bond. In some embodiments, the modified internucleotide bond is a phosphorothioate bond.

[0366] As used herein, "modified nucleotides" refer to nucleotides having one or more chemical modifications compared to a corresponding reference nucleotide selected from the group consisting of: adenine ribonucleotides, guanine ribonucleotides, cytosine ribonucleotides, uracil ribonucleotides, adenine deoxyribonucleotides, guanine deoxyribonucleotides, cytosine deoxyribonucleotides, and thymine deoxyribonucleotides. The modified nucleotides may have, for example, one or more chemical modifications in their sugar, nucleobase, and / or phosphate groups. Additionally, or alternatively, the modified nucleotides may have one or more chemical moieties conjugated to the corresponding reference nucleotides. In some embodiments, the modified nucleotides are 2'-fluoro-modified nucleotides, 2'-O-methyl-modified nucleotides, or 2'-O-alkyl-modified nucleotides. In some embodiments, the modified nucleotides have phosphate analogs, such as 5'-vinylphosphonates. In some embodiments, the modified nucleotides have a base-free portion or a reverse base-free portion, such as the portions shown in Table 3.

[0367] As used herein, the term "SARM1-mediated nervous system disease" refers to a nervous system disease, pathology or injury mediated by SARM1 and / or axonal degeneration.

[0368] As used herein, "nucleotide" refers to an organic compound having a nucleoside (a nucleobase, such as adenine, cytosine, guanine, thymine or uracil, and a pentose, such as ribose or 2'-deoxyribose) linked to a phosphate group. "Nucleotide" can be used as a monomer unit of nucleic acid polymers, such as deoxyribonucleic acid (DNA) and ribonucleic acid (RNA).

[0369] As used herein, "oligonucleotide" refers to a polymer of linked nucleotides, wherein each nucleotide may be modified or unmodified. Oligonucleotides are typically less than about 100 nucleotides in length.

[0370] As used herein, "overhang" means one or more unpaired nucleotides extending from the duplex structure of a double-stranded oligonucleotide. The overhang may include one or more unpaired nucleotides extending from the duplex region at the 5' end or 3' end of the double-stranded oligonucleotide. The overhang may be a 3' or 5' overhang on the antisense strand or sense strand of a double-stranded oligonucleotide.

[0371] As used herein, "patient" refers to a human patient.

[0372] As used herein, "phosphate analog" refers to a chemical moiety that simulates the electrostatic and / or steric properties of a phosphate group. In some embodiments, the phosphate analog is located at the 5' terminal nucleotide of an oligonucleotide, replacing the 5'-phosphate that is typically susceptible to enzymatic removal. The 5' phosphate analog can include a phosphatase-resistant bond. Examples of phosphate analogs include 5' methylene phosphonate (5'-MP) and 5'-(E)-vinyl phosphonate (5'-VP). In some embodiments, the phosphate analog is 5' VP.

[0373] The term "% sequence identity" or "percent sequence identity" relative to a reference nucleic acid sequence is defined as the percentage of nucleotides, nucleosides or nucleobases in a candidate sequence that are identical to the nucleotides, nucleosides or nucleobases in the reference nucleic acid sequence, after optimally aligning the sequences and introducing intervals or overhangs (if necessary) to achieve maximum percent sequence identity. Alignment for purposes of determining percent nucleic acid sequence identity can be accomplished by a variety of means within the skill of the art, such as using publicly available computer software programs, such as those described in Current Protocols in Molecular Biology (Ausubel et al., eds., 1987, Supp. 30, section 7.7.18, Table 7.7.1), and including BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), Clustal W2.0 or Clustal X2.0 software. One skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm required for achieving maximum alignment over the full length of the compared sequence. The percentage of "sequence identity" can be determined by comparing two optimally aligned sequences over a comparison window, wherein the nucleic acid sequence fragment in the comparison window may contain additions or deletions (e.g., gaps or overhangs) compared to the reference sequence (which does not contain additions or deletions) to achieve optimal alignment of the two sequences. The percentage can be calculated by determining the number of positions where the same nucleotide, nucleoside, or nucleobase occurs in the two sequences to give the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison window, and multiplying the result by 100 to give the percentage of sequence identity. The output is the percentage identity of the primary sequence relative to the query sequence.

[0374] As used herein, "RNAi", "RNAi agent", "iRNA", "iRNA agent" and "RNA interference agent" refer to agents that mediate sequence-specific degradation of target mRNA by RNA interference (e.g., via an RNA-induced silencing complex (RISC) pathway). In some embodiments, the RNAi agent has a sense strand and an antisense strand, and the sense strand and antisense strand form a duplex (e.g., double-stranded RNA). In some embodiments, the sense strand has a delivery portion conjugated to the 5' end or 3' end of the sense strand or to the nucleotides of the sense strand.

[0375] As used herein, "strand" refers to a single continuous sequence of nucleotides linked together by internucleotide bonds (eg, phosphodiester bonds or phosphorothioate bonds). A strand may have two free ends (eg, a 5' end and a 3' end).

[0376] As used herein, "SARM1" (sterile alpha and TIR motif protein 1, also known as SARM; HsTIR; SAMD2; hSARM1; MyD88-5) refers to human SARM1 mRNA transcript or human SARM1 protein. The nucleotide sequence of human SARM1 mRNA can be found at NM_015077.4:

[0377]

[0378]

[0379]

[0380]

[0381]

[0382]

[0383] The corresponding amino acid sequence of human SARM1 protein can be found at NP_055892.2:

[0384]

[0385] As used herein, "subject" refers to mammals, including cats, dogs, mice, rats, chimpanzees, apes, monkeys and humans. Preferably, the subject is a human.

[0386] As used herein, "treatment" or "treating" refers to all processes that may slow down, control, delay or stop the progression of the pathology or disease disclosed herein, or improve the symptoms of the pathology or disease, but does not necessarily mean that all symptoms of the pathology or disease are completely eliminated. Treatment includes the administration of proteins or nucleic acids or vectors or compositions to treat a disease or condition in a patient, particularly a human. Example

[0387] Example 1. Synthesis of Linker-Delivery Moiety Pairs

[0388] Certain abbreviations are defined as follows: “ACN” means acetonitrile; “AEX” means anion exchange; “C / D” means cleavage and deprotection; “CPG” means controlled pore glass; “DCM” means dichloromethane; “DEA” means diethylamine; “DIEA” means N,N-diisopropylethylamine; “DMAP” means 4-dimethylaminopyridine; “DMF” means dimethylformamide; “DMSO” means dimethyl sulfoxide; “DMTCl” means 4,4′-dimethoxytrityl chloride; “ES / MS” means electrospray ionization mass spectrometry; “EtOAc” means ethyl acetate; “EtOH” means ethanol and ethyl alcohol; “HBTU” means 3-[bis(dimethylamino)methylinium]-3H-benzotriazole-1-oxide hexafluorophosphate; “HOBt” means 1-hydroxybenzotriazole; “IP-RP” means reverse phase ion pair; “LCAA” means “CPG” refers to long-chain alkylamine controlled-pore glass; “LC / MS” refers to liquid chromatography-mass spectrometry; “MeOH” refers to methanol and methyl alcohol; “MPA” refers to mobile phase A; “MPB” refers to mobile phase B; “MWCO” refers to molecular weight cutoff; “NMR” refers to nuclear magnetic resonance; “PBS” refers to phosphate-buffered saline; “PEG” refers to polyethylene glycol; “PVDF” refers to polyvinylidene fluoride; “RP” refers to reverse phase; “RPM” refers to revolutions per minute; “siRNA” refers to small interfering RNA; “TEA” refers to triethylamine; “THF” refers to tetrahydrofuran; “TLC” refers to thin-layer chromatography; “TMP” refers to 2,2,6,6-tetramethylpiperidine; “UPLC” refers to ultra-performance liquid chromatography; and “UV” refers to ultraviolet light.

[0389] Solution 1

[0390]

[0391] Scheme 1, step A describes the coupling of compounds (1) and (2) using an appropriate base (e.g., DMAP) in a suitable solvent (e.g., DCM) to give compound (3). Step B shows the coupling of compound (3) with 1-amino-3,6,9,12-tetraoxapentadecan-15-oic acid in the presence of a base (e.g., potassium carbonate) and in a solvent system (e.g., water and THF) to give compound (4).

[0392] Solution 2

[0393]

[0394] Scheme 2, step A describes the Mitsunobu reaction of compound (5) with tert-butyl 1-hydroxy-3,6,9,12-tetraoxapentadecan-15-oate using triphenylphosphine and diisopropyl azodicarboxylate in a solvent such as THF to give compound (6). Step B shows the acid deprotection of compound (6) using an acid such as HCl in a solvent such as 1,4-dioxane to give compound (7).

[0395] Solution 3

[0396]

[0397] Scheme 3, step A describes the protection of compound (8) using DMTCl and a suitable base (e.g. DIEA) in a solvent (e.g. DCM) to give compound (9). Step B shows the amide coupling of compound (9) with 4-piperidinylmethanol using HBTU and HOBt with TMP in a solvent (e.g. DCM) to give compound (10). Step C shows the deprotection of compound (10) with 20% piperidine in DMF to give compound (11).

[0398] Solution 4

[0399]

[0400] Scheme 4, step A describes the amide coupling of compound (11) with compound (4) or compound (7) using standard coupling reagents (e.g., HBTU and HOBt) and a base (e.g., DIEA) in a solvent (e.g., DMF) to give compound (12). One skilled in the art will recognize the various conditions that can be used to perform such amide couplings. Step B shows the coupling of compound (12) with succinic anhydride using a base (e.g., TEA) and catalytic DMAP in a solvent (e.g., DCM) to give compound (13). Step C shows the amide coupling of compound (13) with aminoLCAACPG using HBTU and a base (e.g., DIEA) in a solvent (e.g., ACN), followed by a multi-step process to give compound (14).

[0401] Preparation 1

[0402] 2,5-Dioxypyrrolidine-1-palmitate

[0403]

[0404] Palmitic acid (2.00g, 7.80mmol) is added to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.79g, 9.36mmol) and DMAP (0.19g, 1.56mmol) in DCM (31mL) solution.The mixture is stirred at ambient temperature for 5 minutes, then N-hydroxysuccinimide (0.99g, 8.58mmol) is added and stirred at ambient temperature for 18 hours.Vacuum concentration and the obtained crude product is purified by flash chromatography on silica gel, eluted with 0-80% EtOAc gradient in hexane, to obtain the title compound, it is a white solid (2.65g, 96%). 1 H NMR (DMSO-d6) δ2.81 (s, 4H), 2.66 (t, 2H), 1.62 (m, 2H), 1.25 (brs, 24H), 0.87 (t, 3H).

[0405] Preparation 2

[0406] 3-[2-[2-[2-[2-(hexadecanoylamino)ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid

[0407]

[0408] 1-Amino-3,6,9,12-tetraoxapentadecan-15-acid (0.14 g, 0.53 mmol) was added to a solution of potassium carbonate (0.14 g, 1.00 mmol) in THF (1 mL) and water (2 mL). 2,5-dioxopyrrolidine-1-palmitate (0.18 g, 0.51 mmol) was added and the reaction was stirred at ambient temperature for 18 hours. The reaction was quenched with water (30 mL) and the pH was adjusted to ~3 with 1N HCl aqueous solution. A precipitate was formed and collected by vacuum filtration to give the title compound as a white solid (0.19 g, 74%). ES / MS m / z 504 (M+H).

[0409] Preparation 3

[0410] Tert-butyl 3-[2-[2-[2-[2-[(2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-yl]oxyethoxy]ethoxy]ethoxy]ethoxy]propionate

[0411]

[0412] (2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-ol (3.00 g, 6.90 mmol), tert-butyl 1-hydroxy-3,6,9,12-tetraoxapentadecan-15-acid ester (2.50 g, 7.60 mmol) and triphenylphosphine (2.00 g, 7.60 mmol) were mixed in THF (28.0 mL) and diisopropyl azodicarboxylate (1.50 mL, 7.60 mmol) was added dropwise over 5 minutes. The mixture was heated at 60 ° C for 16 hours. The mixture was cooled to ambient temperature, silica gel was added, and concentrated in vacuo to give an off-white solid. The mixture was purified via flash chromatography on silica gel, eluted with 0-40% EtOAc / hexane to give the title compound as an oil (3.33 g, 66%). 1 H NMR (CDCl3): 3.84 (s, 4H), 3.77-3.71 (m, 13H), 2.59 (t, J = 6.8Hz, 2H), 2.52 (t, J = 6.6Hz, 2H), 2.20-2.20 (m, 3H), 2.15- 2.12(m,3H), 2.10(s,3H), 1.87-1.73(m,2H), 1.58-1.51(m,4H), 1.47(s,9H), 1.35-1.27(m,21H), 0.90-0.86(m,12H).

[0413] Preparation 4

[0414] 3-[2-[2-[2-[2-[(2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-yl]oxyethoxy]ethoxy]ethoxy]ethoxy]propanoic acid

[0415]

[0416] Tert-butyl 3-[2-[2-[2-[2-[(2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-yl]oxyethoxy]ethoxy]ethoxy]propanoate (3.33 g, 4.53 mmol) was dissolved in 4M HCl in dioxane (22.6 mL, 90.6 mmol) and stirred at ambient temperature for 16 hours. The solvent was removed under reduced pressure to give the title compound as an off-white solid (3.08 g, 100%). ES / MS m / z 678.0 (MH).

[0417] Preparation 5

[0418] (2S)-3-[Bis(4-methoxyphenyl)-phenyl-methoxy]-2-(9H-fluoren-9-ylmethoxycarbonylamino)propanoic acid

[0419]

[0420] Under an inert atmosphere, DIEA (64 mL, 0.366 mol) was added to a stirred solution of (2S)-2-(9H-fluorene-9-ylmethoxycarbonylamino)-3-hydroxy-propionic acid (40 g, 0.122 mol) in anhydrous DCM (400 mL) at 0 ° C. A solution of DMTCl (49.6 g, 0.146 mol) in DCM (200 mL) was slowly added to the mixture. It was allowed to warm to ambient temperature and stirred for 1 6 hours. Thereafter, the reaction mixture was diluted with water and extracted with DCM. The organic matter was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude residue was washed with 10% EtOAc / hexane and dried under vacuum to obtain the crude title compound as a light brown solid (62 g, crude product). TLC: 5% MeOH / CH2Cl2 (R f :0.5)UV, 254nM.

[0421] Preparation 6

[0422] 9H-Fluoren-9-ylmethyl N-[(1S)-1-[[bis(4-methoxyphenyl)-phenyl-methoxy]methyl]-2-[4-(hydroxymethyl)-1-piperidinyl]-2-oxo-ethyl]carbamate

[0423]

[0424] Under an inert atmosphere, HBTU (78.3 g, 0.206 mol), HOBt (27.9 g, 0.206 mol) and 4-piperidinylmethanol (15.4 g, 0.134 mol) were slowly added to a stirred solution of (2S)-3-[bis(4-methoxyphenyl)-phenyl-methoxy]-2-(9H-fluorene-9-ylmethoxycarbonylamino)propanoic acid (62 g, 0.103 mol) in DCM (750 mL) at 0°C, followed by the addition of TMP (15 mL, 0.113 mol). The resulting reaction mixture was allowed to reach ambient temperature and stirred for 4 hours. Thereafter, the mixture was diluted with water and extracted with DCM. The organics were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified via flash chromatography on silica gel, eluting with 20-40% EtOAc / hexanes and 1% MeOH / DCM to give the title compound (40 g, 52% yield for two steps). 1HNMR (DMSO-d6) δ7.88 (br d, J=7.5Hz, 2H), 7.79-7.59 (m, 3H), 7.45-7.12 (m, 13H), 6.92-6.76 (m, 4H), 4.79-4.44 (m, 2H), 4.32 (br d, J=11.4Hz, 2H), 4.20 (br s, 2H), 3.71 (s, 6H), 3.21 (br s, 4H), 2.99-2.79 (m, 1H), 2.69 (br s, 2H), 1.81-1.43 (m, 3H), 1.08-0.73 (m, 2H).

[0425] Preparation 7

[0426] (2S)-2-Amino-3-[bis(4-methoxyphenyl)-phenyl-methoxy]-1-[4-(hydroxymethyl)-1-piperidinyl]propan-1-one

[0427]

[0428] Under an inert atmosphere, a solution of 20% piperidine in DMF (400 mL) was slowly added to 9H-fluorene-9-ylmethyl N-[(1S)-1-[[bis(4-methoxyphenyl)-phenyl-methoxy]methyl]-2-[4-(hydroxymethyl)-1-piperidinyl]-2-oxo-ethyl]carbamate (40 g, 0.055 mol) at 0°C. The mixture was allowed to warm to ambient temperature and stirred for 1 hour. Thereafter, the mixture was diluted with water and extracted with EtOAc. The organics were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified via flash chromatography on silica gel, eluted with 1-8% MeOH / DCM to give the title compound as an off-white solid (13 g, 47%). ES / MS m / z 1009.5 (2M+H).

[0429] Preparation 8

[0430] N-[2-[2-[2-[2-[3-[[(1S)-1-[[bis(4-methoxyphenyl)-phenyl-methoxy]methyl]-2-[4-(hydroxymethyl)-1-piperidinyl]-2-oxo-ethyl]amino]-3-oxo-propoxy]ethoxy]ethoxy]ethoxy]ethyl]hexadecanamide

[0431]

[0432] 3-[2-[2-[2-[2-[2-(hexadecanoylamino)ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (496 mg, 0.984 mmol), HOBt (146 mg, 1.08 mmol), HBTU (410 mg, 1.08 mmol) and DIEA (1.03 mL, 5.90 mmol) were mixed in DMF (9.84 mL) and stirred at ambient temperature for 10 minutes. (2S)-2-amino-3-[bis(4-methoxyphenyl)-phenyl-methoxy]-1-[4-(hydroxymethyl)-1-piperidinyl]propan-1-one (546 mg, 1.08 mmol) was added to the mixture and stirred at ambient temperature for 16 hours. The mixture was layered between EtOAc and saturated sodium chloride aqueous solution. The layers were separated and the organics were washed with saturated sodium chloride aqueous solution. The organics were dried over sodium sulfate, filtered, and concentrated in vacuo. The resultant residue was purified by flash chromatography on silica gel eluting with 0-10% MeOH / DCM to give the title compound as an oil (327 mg, 34%). 1 H NMR (DMSO-d6) 8.21 (d, J=8.5Hz, 1H), 7.80 (t, J=5.6Hz, 1H), 7.37-7.28 (m, 4H), 7.23-7.20 (m, 5H), 6.88 (d, J=8.3H z, 4H), 5.06-5.02 (m, 1H), 4.51-4.49 (m, 1H), 4.45-4.40 (m, 1H), 3.97-3.93 (m, 1H), 3.74 (s, 5H), 3.63-3.56 (m, 2H) , 3.49-3.48(m, 4H), 3.47-3.45(m, 7H), 3.40-3.35(m, 2H), 3.30(s, 1H), 3.23-3.13(m, 7H), 2.41-2.33(m, 2H), 2.04 (t, J=7.4Hz, 2H), 1.74-1.69 (m, 3H), 1.51-1.44 (m, 2H), 1.26-1.24 (m, 24H), 1.00-0.97 (m, 1H), 0.88-0.82 (m, 5H).

[0433] Preparation 9

[0434] 4-[[1-[(2S)-3-[bis(4-methoxyphenyl)-phenylmethoxy]-2-[3-[2-[2-[2-[2-(hexadecanoylamino)ethoxy]ethoxy]ethoxy]propionylamino]propionyl]-4-piperidinyl]methoxy]-4-oxobutanoic acid

[0435]

[0436] N-[2-[2-[2-[2-[3-[[(1S)-1-[[bis(4-methoxyphenyl)-phenylmethoxy]methyl]-2-[4-(hydroxymethyl)-1-piperidinyl]-2-oxoethyl]amino]-3-oxopropoxy]ethoxy]ethoxy]ethoxy]ethyl]hexadecanamide (320 mg, 0.323 mmol), DMAP (120 mg, 0.969 mmol), TEA (225 μL, 1.62 mmol) and succinic anhydride (64.7 mg, 0.646 mmol) were mixed in DCM (6.46 mL), and the mixture was stirred at ambient temperature for 16 hours. The mixture was purified directly by flash chromatography on silica gel, eluting with 0% to 40% MeOH / DCM to give the title compound as a colorless oil (279 mg, 79%). 1 H NMR (DMSO-d6) 12.65-12.64 (m, 1H), 8.24-8.19 (m, 1H), 7.80 (t, J=5.6Hz, 1H), 7.37-7.28 (m, 4H), 7.2 4-7.20 (m, 5H), 6.88 (d, J=8.6Hz, 4H), 5.05-5.01 (m, 1H), 4.44-4.40 (m, 1H), 3.97-3.95 (m, 3H), 3.74 ( s, 6H), 3.61-3.56 (m, 2H), 3.49-3.45 (m, 11H), 3.38 (t, J=5.9Hz, 3H), 3.22-3.14 (m, 6H), 2.48-2.31 ( m, 7H), 2.04 (t, J=7.4Hz, 2H), 1.90-1.87 (m, 5H), 1.24 (s, 23H), 0.98-0.96 (m, 1H), 0.87-0.82 (m, 4H).

[0437] Preparation 10

[0438] 4-[[1-[(2S)-3-[bis(4-methoxyphenyl)-phenyl-methoxy]-2-[3-[2-[2-[2-[(2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-yl]oxyethoxy]ethoxy]ethoxy]ethoxy]propionylamino]propionyl]-4-piperidinyl]methoxy]-4-oxo-butyric acid

[0439]

[0440] 3-[2-[2-[2-[2-[(2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-yl]oxyethoxy]ethoxy]ethoxy]propanoic acid (1.20 g, 1.80 mmol), HOBt (260 mg, 1.90 mmol), HBTU (740 mg, 1.90 mmol) and DIEA (1.80 mL, 11.0 mmol) were mixed in DMF (18.0 mL) and stirred at ambient temperature for 10 minutes. To the mixture was added (2S)-2-amino-3-[bis(4-methoxyphenyl)-phenyl-methoxy]-1-[4-(hydroxymethyl)-1-piperidinyl]propan-1-one (980 mg, 1.90 mmol) and stirred at ambient temperature for 16 hours. The mixture was partitioned between EtOAc and saturated aqueous sodium chloride solution. The layers were separated and the organic matter was washed with saturated sodium chloride aqueous solution. The organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by flash chromatography on silica gel, eluted with 0-10% MeOH / DCM to give N-[(1S)-1-[[bis(4-methoxyphenyl)-phenyl-methoxy]methyl]-2-[4-(hydroxymethyl)-1-piperidinyl]-2-oxo-ethyl]-3-[2-[2-[2-[(2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-yl]oxyethoxy]ethoxy]ethoxy]ethoxy]acrylamide as a yellow oil.

[0441] N-[(1S)-1-[[Bis(4-methoxyphenyl)-phenyl-methoxy]methyl]-2-[4-(hydroxymethyl)-1-piperidinyl]-2-oxo-ethyl]-3-[2-[2-[2-[2-[(2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-yl]oxyethoxy]ethoxy]ethoxy]ethoxy]acrylamide (1.45 g, 1.24 mmol), DMAP (456 mg, 3.73 mmol), TEA (867 μL, 6.22 mmol) and succinic anhydride (249 mg, 2.49 mmol) were combined in DCM (24.9 mL) and stirred at ambient temperature for 16 hours. Concentrate in vacuo and purify the resulting residue via flash chromatography on silica gel eluting with 0-40% MeOH / DCM to give the title compound (1.36 g, 60%) as an oil. ES / MS m / z 1264.4 (MH).

[0442] Preparation 11

[0443] 4-[[1-[(2S)-3-[bis(4-methoxyphenyl)-phenyl-methoxy]-2-[3-[2-[2-[2-[2-(hexadecanoylamino)ethoxy]ethoxy]ethoxy]propionylamino]propionyl]-4-piperidinyl]methoxy]-4-oxo-butyryl]amino] (CPG)

[0444]

[0445] 4-[[1-[(2S)-3-[bis(4-methoxyphenyl)-phenyl-methoxy]-2-[3-[2-[2-[2-[2-(hexadecanoylamino)ethoxy]ethoxy]ethoxy]propionamido]propionyl]-4-piperidinyl]methoxy]-4-oxo-butyric acid (270 mg, 0.248 mmol) was dissolved in ACN (12.5 mL) and the solution was transferred to a sintered glass dropping funnel. DIEA (150 μL, 0.860 mmol) and HBTU (190 mg, 0.500 mmol) were added to the solution and the mixture was shaken at ambient temperature for 10 minutes. Natural amino LCAA CPG was added to the solution. =(1.92g, 129μmol / g), and at ambient temperature with 500RPM vibration mixture 16 hours. Drain CPG and dry under nitrogen for 5 minutes. Use DCM (50mL), 10%MeOH / DCM (50mL) and then wash CPG with ether (50mL). Dry CPG 30 minutes under nitrogen, and resuspend in pyridine (15mL) subsequently. Add acetic anhydride (3.30mL, 35.0mmol) and TEA (0.50mL), at ambient temperature with 500RPM vibration mixture 2 hours. Drain CPG and dry under nitrogen for 5 minutes. Use DCM (50mL), 10%MeOH / DCM (50mL) and then wash CPG with ether (50mL). Dry CPG 45 minutes under nitrogen, and determine the ligand load at 505nm, to obtain title compound (1.92g, 75.5μmol / g).

[0446] Preparation 12

[0447] [[4-[[1-[(2S)-3-[bis(4-methoxyphenyl)-phenyl-methoxy]-2-[3-[2-[2-[2-[(2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-yl]oxyethoxy]ethoxy]ethoxy]ethoxy]propionylamino]propionyl]-4-piperidinyl]methoxy]-4-oxo-butyryl]amino] (CPG)

[0448]

[0449] The title compound was prepared from 4-[[1-[(2S)-3-[bis(4-methoxyphenyl)-phenyl-methoxy]-2-[3-[2-[2-[2-[2-[(2R)-2,5,7,8-tetramethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]chroman-6-yl]oxyethoxy]ethoxy]ethoxy]ethoxy]propionylamino]propionyl]-4-piperidinyl]methoxy]-4-oxo-butanoic acid in a manner substantially similar to that of the preparation of 11. The ligand loading was determined at 505 nm to give the title compound (4.01 g, 66.9 μmol / g).

[0450] Preparation 13

[0451] 3-[[(2R,3R,4R,5R)-2-[[bis(4-methoxyphenyl)-phenylmethoxy]methyl]-4-hexadecyl-5-(2-hydroxy-4-oxo-pyrimidin-1-yl)THF-3-yl]oxy-(diisopropylamino)phosphoryl]oxypropionitrile

[0452]

[0453] The title compound was prepared according to the protocol described in WO2019217459. 1 H NMR(CD3CN): 7.86-7.73(m, 1H), 7.51-7.43(m, 2H), 7.40-7.23(m, 7H), 6.95-6.87( m, 4H), 5.90-5.84 (m, 1H), 5.29-5.21 (m, 1H), 4.54-4.40 (m, 1H), 4.21-4.13 (m, 1H), 4.10-3.56(m, 13H), 3.50-3.34(m, 2H), 2.75-2.62(m, 1H), 2.55(t, J=6.0Hz, 1H), 1 .66-1.51 (m, 2H), 1.40-1.14 (m, 35H), 1.08 (d, J=6.8Hz, 3H), 0.91 (t, J=6.8Hz, 3H). 31 P NMR(CD3CN): 149.6, 149.2.

[0454] Preparation 14

[0455] N-[9-[(2R,3R,4R,5R)-5-[[Bis(4-methoxyphenyl)-phenylmethoxy]methyl]-4-[2-cyanoethoxy-(diisopropylamino)phosphoryl]oxy-3-hexadecyloxy-tetrahydrofuran-2-yl]purin-6-yl]benzamide

[0456]

[0457] The title compound was prepared according to the protocol described in WO2019217459. 1 H-NMR (CD3CN) δ9.37 (s, 1H), 8.57 (d, J = 9.4Hz, 1H), 8.27 (d, J = 10.3Hz, 1H), 7.99 (d, J = 7.6Hz, 2H), 7.61 (d, J = 7.4Hz, 1H), 7.52 (t, J=7.6Hz, 2H), 7.42 (t, J=7.3Hz, 2H), 7.34-7.16 (m, 7H), 6.85-6.77 (m, 4H), 6.11 (dd, J=5.0, 2.5Hz, 1H), 4.80 (m, 1 H), 4.69 (m, 1H), 4.32 (m, 1H), 3.97-3.78 (m, 1H), 3.74 (d, J=3.1Hz, 7H), 3.64 (m, 4H), 3.56-3.40 (m, 2H), 3.33 (m, 1H), 2.73 -2.59 (m, 1H), 2.50 (t, J=6.0Hz, 1H), 1.52-1.45 (m, 2H), 1.33-1.12 (m, 37H), 1.09 (d, J=6.8Hz, 3H), 0.87 (t, J=6.8Hz, 3H). 31 P NMR (CD3CN) δ 151.19, 150.78.

[0458] Preparation 15

[0459] N-[1-[(2R,3R,4R,5R)-5-[[Bis(4-methoxyphenyl)-phenylmethoxy]methyl]-4-[2-cyanoethoxy-(diisopropylamino)phosphoryl]oxy-3-hexadecyloxy-tetrahydrofuran-2-yl]-2-oxo-pyrimidin-4-yl]acetamide

[0460]

[0461] The title compound was prepared according to the protocol described in WO2019217459. 1H-NMR (CD3CN) 9.15 (s, 1H), 8.46 (dd, J=7.5Hz, 1H), 7.95 (d, J=7.6Hz, 2H), 7.63 (t, J=7.5Hz, 1H), 7.57-7.41 (m, 5H), 7.41-7.31 (m , 6H), 7.28 (m, 1H), 7.04 (d, J=15.8Hz, 1H), 6.90 (t, J=7.9Hz, 4H), 5.90 (d, J=7.8Hz, 1H), 4.51 (m, 1H), 4.20 (dd, J=10.6, 8.1Hz, 1H) , 4.04 (dd, J=31.3, 4.6Hz, 1H), 3.91-3.81 (m, 2H), 3.79 (d, J=3.1Hz, 6H), 3.74 (m, 2H), 3.69-3.41 (m, 6H), 2.67-2.59 (m, 1H), 2.54 -2.48 (m, 1H), 1.58 (m, 2H), 1.36 (m, 2H), 1.25 (d, J = 4.7Hz, 26H), 1.21-1.09 (m, 10H), 1.04 (d, J = 6.8Hz, 3H), 0.87 (t, J = 6.8Hz, 3H). 31 P NMR (CD3CN) δ 151.10, 150.19.

[0462] Preparation 16

[0463] N-[9-[(2R,3R,4R,5R)-5-[[Bis(4-methoxyphenyl)-phenylmethoxy]methyl]-4-[2-cyanoethoxy-(diisopropylamino)phosphoryl]oxy-3-hexadecyloxy-tetrahydrofuran-2-yl]-6-oxo-1H-purin-2-yl]-2-methyl-propionamide

[0464]

[0465] The title compound was prepared according to the protocol described in WO2019217459. 1H-NMR (CDCl3) δ12.01-11.96 (m, 1H), 7.82-7.78 (m, 1H), 7.59-7.53 (m, 1H), 7.47-7.42 (m, 1H), 7.41-7.37 (m, 2H), 7.34-7.29(m, 2H), 7.27-7.22(m, 3H), 6.85-6.80(m, 4H), 5.99-5.82(m, 1H), 4.40-4.36(m, 1H), 4.17-4. 11(m, 1H), 3.80-3.77(m, 6H), 3.76-3.68(m, 6H), 3.22-3.17(m, 1H), 2.84-2.79(m, 1H), 1.60-1.54(m, 4H), 1. 35-1.30(m, 6H), 1.27(s, 19H), 1.24-1.15(m, 13H), 1.06-1.03(m, 5H), 0.93-0.88(m, 6H), 0.74-0.70(m, 1H). 31 P NMR (CDCl3) δ150.20.149.92.

[0466] Preparation 17

[0467] 4-[[1-[(2S)-2-[6-[[(2S)-2-[5-[3-acetylamino-4,5-diacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]oxypentanoylamino]-5-[bis[2-[5-[3-acetylamino-4,5-diacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]oxypentanoylamino]ethyl]amino]-5-oxo-pentanoyl]amino]hexanoylamino]-3-[bis(4-methoxyphenyl)-phenyl-methoxy]propionyl]-4-piperidinyl]methoxy]-4-oxo-butyric acid

[0468]

[0469] The title compound was prepared according to the protocol described in WO2022271806. ES / MS m / z 1109.60 (M-2H) / 2.

[0470] Preparation 18

[0471] Resin loading

[0472]

[0473] According to the method described in WO2022271806, the title compound was prepared by 4-[[1-[(2S)-2-[6-[[(2S)-2-[5-[3-acetylamino-4,5-diacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]oxypentanoylamino]-5-[bis[2-[5-[3-acetylamino-4,5-diacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]oxypentanoylamino]ethyl]amino]-5-oxo-pentanoyl]amino]hexanoylamino]-3-[bis(4-methoxyphenyl)-phenyl-methoxy]propionyl]-4-piperidinyl]methoxy]-4-oxo-butyric acid. The resin loading was calculated to be 34.7 μmol / g.

[0474] Example 2. Synthesis of SARM1 RNAi Agents and Conjugated RNAi Agents

[0475] Via MerMade TM Single strands (sense and antisense) of RNA duplexes were synthesized on a solid support using 12 (LGC Biosearch Technologies). The sequences of the sense and antisense strands are shown in Table 2. Oligonucleotides were synthesized via phosphoramidite chemistry at a scale of 5, 10, 25, or 50 μmol.

[0476] For the sense strand, the types of solid supports were universal CPG: (3'-piperidinol-PEG-palmitate) and (3'-piperidinol-PEG-tocopherol), which were synthesized in-house (see Example 1), while Universal UnyLinker (Chemgenes, Catalog No. AT273-27), 3'-Teg-tocopherol (LGC Biosearch Technologies, Catalog No. BG7-1190) and 3'-cholesterol-TEG CPG (Glen Research, Catalog No. 20-2975) were commercially available. For all antisense strands, commercially available standard support mA was utilized. Standard reagents (Table 7) were used in oligonucleotide synthesis, wherein 0.1 M hydrogenated xanthine in pyridine was used as a sulfurizing agent, and 20% DEA in ACN was used as a post-synthesis auxiliary detergent. All monomers (Table 8) were made at 0.1 M in ACN and contained molecular sieve capture bags.

[0477] Oligonucleotides were cleaved and deprotected (C / D) at 45°C for 20 hours. Ammonia (28-30%, cold) was used to C / D the sense strand from CPG, while 3% DEA in ammonia (28-30%, cold) was used for the antisense strand. C / D was determined to be complete when the mass data confirmed the sequence identity by IP-RP LCMS. CPG was filtered through a 0.45 μm PVDF syringeless filter, a 0.22 μm PVDF filter, and a 10 μm PVDF filter cartridge, depending on the scale. Vacuum filtration or 0.22μm PVDF Quick release was used for filtration. The CPG was backwashed / rinsed with 30% ACN / RNA-free water or 30% EtOH / RNA-free water, then filtered through the same filtration unit and combined with the first filtrate. This was repeated twice. The material was then evenly divided into 50 mL falcon tubes for filtration via Genevac TM After concentration, the crude oligonucleotides were diluted back to synthesis scale with RNAse-free water and filtered through a 0.45 μm PVDF syringe-free filter, a 0.22 μm PVDF Vacuum filtration or 0.22μm PVDF Quick release for filtration.

[0478] Use anion exchange (AEX) or reverse phase (RP) (such as source 15Q-RP column) via AKTA TM Pure purification system purifies crude oligonucleotides. For AEX, ES Industry Source TM The column temperature of the 15Q column was maintained at 65°C with MPA: 20 mM NaH2PO4, 15% ACN, pH 7.4 and MPB: 20 ​​mM NaH2PO4, 1 M NaBr, 15% ACN, pH 7.4. For RP, Source TM 15Q-RP column with MPA: 50 mM NaOAc and 10% CAN and MPB: 50 mM NaOAc and 80% ACN. In all cases, fractions with a mass purity higher than 85% and impurity content not higher than 5% were pooled.

[0479] The purified oligonucleotides were desalted at 3500xg for ~30 minutes using a 15mL 3K MWCO centrifuge tube. The oligonucleotides were rinsed with RNAse-free water until the conductivity of the eluent was less than 100usemi / cm. After desalting, 2-3mL of RNAse-free water was added, followed by 10 aspirations, and the residue was transferred to a 50mL falcon tube, and the above was repeated until the concentration of the compound on the filter was measured by nanodrop and the oligonucleotide was completely transferred. The final oligonucleotide was nanofiltered twice by centrifugation at 3500xg for 2 minutes in a 15mL 100K MWCO centrifuge tube. The final desalted oligonucleotides were analyzed for concentration (nanodrop A260), and the mass purity was characterized by IP-RP LCMS and the UV purity was determined by UPLC.

[0480] To prepare duplexes, equimolar amounts of sense and antisense strands were combined and heated at 65°C for 10 minutes, followed by slow cooling to ambient temperature over 40 minutes. The integrity of the duplexes was confirmed by UPLC analysis and characterized by LCMS using IP-RP. All duplexes were nanofiltered and then filtered through Charles River Endotoxin levels were measured by Cartridge Device to obtain the final RNAi conjugated compound (Table 9). For in vivo analysis, appropriate amounts of duplexes were lyophilized and subsequently reconstituted in IX PBS for rodent studies and in CSF for non-human primate studies.

[0481] Table 7 - Oligonucleotide synthesis reagents

[0482] Reagents Activator solution (0.5M ETT in ACN) Cap A (acetic anhydride, pyridine, dissolved in THF, 1:1:8) Cap B (1-methylimidazole, dissolved in THF, 16:84) Oxidizing solution (0.02M iodine, dissolved in THF / pyridine / water, 70:20:10) Deblocking solution, 3% TCA in DCM (w / v) Acetonitrile (anhydrous solvent, water content up to 10 ppm) Hydrogenated xanthan gum (0.1M, dissolved in pyridine) Diethylamine (20%, dissolved in acetonitrile)

[0483] Table 8 - Phosphoramidites

[0484]

[0485] Table 9 - Conjugated SARM1 RNAi Reagents

[0486]

[0487]

[0488]

[0489]

[0490] "S" means sense strand; "AS" means antisense strand. *LDP 1, 8, 9 are attached to the 3' end of the sense strand. AS SEQ ID NOs: 14, 16, 18, 20, 22, and 24 include 5'-vinylphosphonate; and all other AS in this table include 5'-phosphate.

[0491] Example 3. Characterization of SARM1 RNAi Reagents

[0492] The selected SARM1 RNAi agents were tested in vitro for SARM1 inhibition in cultured cells, including SH-SY5Y cells and / or human induced pluripotent stem cells (hiPSCs).

[0493] Materials and methods

[0494] SH-SY5Y cell culture and RNAi treatment and analysis: SH-SY5Y cells (ATCC CRL-2266) are derived from the SK-N-SH neuroblastoma cell line (Ross, RA et al., 1983. J Natl Cancer Inst 71, 741-747). The basal medium consists of a 1:1 mixture of Eagle's minimum essential medium (Cat No. 30-2003) and F12 medium prepared by ATCC. The complete growth medium was supplemented with 10% fetal bovine serum, 1X amino acids, 1X sodium bicarbonate and 1X penicillin-streptomycin (Gibco), and the cells were incubated in a 5% CO2 humidified environment at 37°C. On the first day, SH-SY5Y cells were plated in fibronectin-coated 96-well tissue culture plates and allowed to attach overnight. On the second day, the complete medium was removed and replaced with RNAi reagents in serum-free medium. The cells were incubated with RNAi reagents for 72 hours, followed by analysis of gene expression. Using Cells-to-C TKit measures the analysis change of gene expression in SH-SY5Y cells processed by RNAi according to the manufacturer's scheme (ThermoFisher A35377).Pre-designed gene expression assay (provided as 20X mixture) is selected from Applied Bio-systems (Foster City, CA, USA).The efficiency of these assays (ThermoFisher Hs00240906_m1SARM1 and ThermoFisher Hs99999905_m1 GAPDH) is characterized by cDNA serial dilutions.RT-QPCR is carried out using QuantStudio 7 Flex system in MicroAmpOptical 384-well reaction plates.The housekeeping gene GAPDH is normalized using delta-delta CT method to determine the relative amount of gene expression.IC50 is determined by four-parameter logistic fitting using GraphPad Prism v9.0.

[0495] Human induced pluripotent stem cell derived neurons (hiPSC neurons) culture and RNAi treatment and analysis: Bioneer developed doxycycline-inducible neuroelement 2 (NGN2) human induced pluripotent stem cells (hiPSCs) for Eli Lilly. hiPSCs were induced with doxycycline for three days (DIV3) to initiate neuronal differentiation and were plated at 30k / well on 96-well PDL and laminin coated plates and grown in an incubator (37°C / 5% CO2) in a neuronal differentiation medium (NDM) consisting of DMEM / F12 (Life Technologies 11330-057), Neurobasal medium (Gibco 15240062), antibiotics, supplements, growth factors and doxycycline. Cells were half-fed every seven days, and at DIV21, the RNAi reagent was serially diluted in NDM and the cells were treated with RNAi, where 75 mL was aspirated and 75 mL of 2xRNAi concentration was added according to the dilution to finally obtain 1x RNAi. After treatment, cells were half-fed every seven days by removing half of the medium and re-adding fresh NDM. Cell lysates were harvested at DIV35 (14 days after RNAi treatment) or DIV42 (21 days after RNAi treatment) and RT-qPCR was performed using TaqMan Fast Advanced Cells-to-CT Kit (ThermoFisher, A35377) and mRNA knockdown was determined using the SARM1 probe as the target gene (ThermoFisher, Hs00240907_m1) and the ACTb probe as the housekeeping gene (ThermoFisher, Hs99999903_m1).

[0496] result

[0497] The percentage knockdown of SARM1 mRNA and IC50 of SARM RNAi agents in human SH-SY5Y cells are summarized in Table 10. The SARM1 RNAi agents tested achieved potent knockdown of SARM1 mRNA in human SH-SY5Y cells.

[0498] Table 10. In vitro activities of selected SARM1 RNAi agents

[0499]

[0500] "NA" means not available.

[0501] Example 4. Knockdown of SARM1 mRNA in cynomolgus monkeys following a single intrathecal administration of selected RNAi agents

[0502] The efficacy of the selected SARM1 RNAi reagents was tested in cynomolgus monkeys (Macaca fascicularis). In order to evaluate the efficacy of RNAi reagents in knocking down SARM1; Five groups of four cynomolgus monkeys were inserted with indwelling catheters at L4 / L5, with the tip of the catheter placed at T12 / L1. Monkeys were infused with aCSF (cerebrospinal fluid) or 20mg conjugated RNAi reagent No.1-4 (8mg / ml in aCSF) over 15 minutes and perfused after 28 days. Tissues collected at autopsy included spinal cord (lumbar, cervical, thoracic) and brain (motor cortex, medulla oblongata, pons and midbrain). qPCR was performed to determine the mRNA knockdown in multiple spinal cord and brain regions.

[0503] The percentage knockdown of SARM1 mRNA in cynomolgus monkeys is summarized in Table 11. As shown in Table 11, potent knockdown of SARM1 mRNA was observed in all tested regions 28 days after a single intrathecal administration of the RNAi agent.

[0504] Table 11. Percent knockdown (KD) of SARM1 mRNA in cynomolgus monkeys following a single intrathecal administration of selected RNAi agents.

[0505]

[0506] Example 5. Knockdown of human SARM1 mRNA in the liver of AAV mouse model

[0507] All animals were housed individually in a temperature-controlled facility with a 12 h / 12 ​​h light / dark cycle. The animal protocol in this study was approved by the Animal Use and Care Committee of Eli Lilly and Co. (protocol number 20-025). C57BL / 6 male mice aged ∼8 weeks were used and a single dose of 10 μg / mL of 10 μg / mL of PBS was administered intravenously. 11 A liver-centered model overexpressing human SARM1 was generated by using an AAV8-TBG vector containing Homo sapiens SARM1 with 100 genome copies (GC). 14 days after AAV administration, the selected RNAi agent or control agent (PBS) was injected subcutaneously at 3 mg / kg (n=6 per group). 14 days after subcutaneous injection of RNAi agent or control, mice were sacrificed and the level of human SARM1 mRNA in the liver was evaluated. The level of knockdown of human SARM1 mRNA in vivo was determined by comparison with the control group.

[0508] Table 12 summarizes the percent knockdown of human SARM1 mRNA in the liver of the AAV mouse model.

[0509] Table 12. Percent knockdown (KD) of human SARM1 mRNA in the liver of the AAV mouse model.

[0510]

[0511]

[0512] Sequence Listing

[0513]

[0514]

[0515]

[0516]

[0517]

[0518]

Claims

1. A SARM1 RNAi agent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, Wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the following: (a) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 2; (b) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 3, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 4; (c) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:5, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:6; (d) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:7, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO:8; (e) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 9, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 10; and (f) the sense strand comprises a first nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 11, and the antisense strand comprises a second nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 12, wherein optionally, one or more nucleotides of the sense strand and the antisense strand are independently modified nucleotides, and Optionally, one or more internucleotide bonds of the sense strand and the antisense strand are modified internucleotide bonds.

2. The SARM1 RNAi agent of claim 1, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the group consisting of: (a) the sense strand comprises a first nucleic acid sequence SEQ ID NO: 1, and the antisense strand comprises a second nucleic acid sequence SEQ ID NO: 2; (b) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 3, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 4; (c) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 5, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 6; (d) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 7, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 8; (e) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 9, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 10; and (f) the sense strand comprises a first nucleic acid sequence of SEQ ID NO:11, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO:

12.

3. The SARM1 RNAi agent of claim 1 or 2, wherein the sense strand and the antisense strand have a pair of nucleic acid sequences selected from the following: (a) the sense strand has a first nucleic acid sequence of SEQ ID NO: 1, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 2; (b) the sense strand has a first nucleic acid sequence of SEQ ID NO: 3, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 4; (c) the sense strand has a first nucleic acid sequence of SEQ ID NO: 5, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 6; (d) the sense strand has a first nucleic acid sequence of SEQ ID NO: 7, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 8; (e) the sense strand has a first nucleic acid sequence of SEQ ID NO: 9, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 10; and (f) The sense strand has a first nucleic acid sequence of SEQ ID NO:11, and the antisense strand has a second nucleic acid sequence of SEQ ID NO:

12.

4. The SARM1 RNAi agent of any one of claims 1 to 3, wherein one or more nucleotides of the sense strand are modified nucleotides.

5. The SARM1 RNAi agent of any one of claims 1 to 4, wherein each nucleotide of the sense strand is a modified nucleotide.

6. The SARM1 RNAi agent of any one of claims 1-5, wherein one or more nucleotides of the antisense strand are modified nucleotides.

7. The SARM1 RNAi agent of any one of claims 1-6, wherein each nucleotide of the antisense strand is a modified nucleotide.

8. The SARM1 RNAi agent of any one of claims 1-7, wherein the modified nucleotide is a 2'-fluoro-modified nucleotide, a 2'-O-methyl-modified nucleotide, or a 2'-O-alkyl-modified nucleotide.

9. The SARM1 RNAi agent of any one of claims 1 to 8, wherein the sense strand has four 2'-fluoro-modified nucleotides at positions 7, 9, 10, and 11 from the 5' end of the sense strand.

10. The SARM1 RNAi agent of claim 9, wherein the nucleotides at positions other than positions 7, 9, 10, and 11 on the sense strand are 2'-O-methyl modified nucleotides.

11. The SARM1 RNAi agent of any one of claims 8 to 10, wherein the antisense strand has four 2'-fluoro-modified nucleotides at positions 2, 6, 14, and 16 from the 5' end of the antisense strand.

12. The SARM1 RNAi agent of claim 11, wherein the nucleotides at positions other than positions 2, 6, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides.

13. The SARM1 RNAi agent of any one of claims 1-8, wherein the sense strand has three 2'-fluoro-modified nucleotides at positions 9, 10, and 11 from the 5' end of the sense strand.

14. The SARM1 RNAi agent of claim 13, wherein the nucleotides at positions other than positions 9, 10, and 11 on the sense strand are 2′-O-methyl modified nucleotides.

15. The SARM1 RNAi agent of any one of claims 1-10, 13, 14, wherein the antisense strand has five 2'-fluoro-modified nucleotides at positions 2, 5, 7, 14, and 16 from the 5' end of the antisense strand.

16. The SARM1 RNAi agent of claim 15, wherein the nucleotides at positions other than positions 2, 5, 7, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides.

17. The SARM1 RNAi agent of any one of claims 1-10, 13, 14, wherein the antisense strand has five 2'-fluoro-modified nucleotides at positions 2, 5, 8, 14, and 16 from the 5' end of the antisense strand.

18. The SARM1 RNAi agent of claim 17, wherein the nucleotides at positions other than positions 2, 5, 8, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides.

19. The SARM1 RNAi agent of any one of claims 1-10, 13, 14, wherein the antisense strand has five 2'-fluoro-modified nucleotides at positions 2, 3, 7, 14, and 16 from the 5' end of the antisense strand.

20. The SARM1 RNAi agent of claim 19, wherein the nucleotides at positions other than positions 2, 3, 7, 14, and 16 on the antisense strand are 2'-O-methyl modified nucleotides.

21. The SARM 1 RNAi agent of any one of claims 1-10, 13, and 14, wherein the antisense strand has two 2'-fluoro-modified nucleotides at the 2nd and 14th positions from the 5' end of the antisense strand.

22. The SARM1 RNAi agent of claim 21, wherein the nucleotides at positions other than positions 2 and 14 on the antisense strand are 2'-O-methyl modified nucleotides.

23. The SARM1 RNAi agent of any one of claims 1-22, wherein the sense strand and the antisense strand have one or more modified internucleotide bonds.

24. The SARM1 RNAi agent of claim 23, wherein the modified internucleotide bond is a phosphorothioate bond.

25. The SARM1 RNAi agent of claim 24, wherein the sense strand has four or five phosphorothioate bonds.

26. The SARM1 RN Ai agent of claim 24 or 25, wherein the antisense strand has four or five phosphorothioate bonds.

27. The SARM1 RNAi agent of any one of claims 1-26, wherein the first nucleotide from the 5' end of the antisense strand is a modified nucleotide having a phosphate analog.

28. The SARM1 RNAi agent of claim 27, wherein the phosphate analog is 5'-vinyl phosphonate. E 29. The SARM1 RNAi agent of any one of E 1-28, wherein the sense strand comprises an abasic portion or an inverted abasic portion.

30. A SARM1 RNAi reagent having a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a duplex, Wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from the following: (a) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 14; (b) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 16; (c) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 18; (d) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 20; (e) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 22; (f) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 24; (g) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 27; (h) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 29; (i) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 30; (j) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 31; (k) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 32; (l) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 33; (m) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 35; (n) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 36; (o) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 37; (p) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 38; (q) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 39; (r) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 41; (s) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 42; (t) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 43; (u) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 44; (v) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 45; (w) the sense strand comprises a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO: 46; and (x) The sense strand comprises a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand comprises a second nucleic acid sequence of SEQ ID NO:

47.

31. The SARM1 RNAi agent of claim 30, wherein the sense strand and the antisense strand have a pair of nucleic acid sequences selected from the group consisting of: (a) the sense strand has a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 14; (b) the sense strand has a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 16; (c) the sense strand has a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 18; (d) the sense strand has a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 20; (e) the sense strand has a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 22; (f) the sense strand has a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 24; (g) the sense strand has a first nucleic acid sequence of SEQ ID NO: 15, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 27; (h) the sense strand has a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 29; (i) the sense strand has a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 30; (j) the sense strand has a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 31; (k) the sense strand has a first nucleic acid sequence of SEQ ID NO: 28, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 32; (l) the sense strand has a first nucleic acid sequence of SEQ ID NO: 13, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 33; (m) the sense strand has a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 35; (n) the sense strand has a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 36; (o) the sense strand has a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 37; (p) the sense strand has a first nucleic acid sequence of SEQ ID NO: 34, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 38; (q) the sense strand has a first nucleic acid sequence of SEQ ID NO: 21, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 39; (r) the sense strand has a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 41; (s) the sense strand has a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 42; (t) the sense strand has a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 43; (u) the sense strand has a first nucleic acid sequence of SEQ ID NO: 40, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 44; (v) the sense strand has a first nucleic acid sequence of SEQ ID NO: 17, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 45; (w) the sense strand has a first nucleic acid sequence of SEQ ID NO: 19, and the antisense strand has a second nucleic acid sequence of SEQ ID NO: 46; and (x) The sense strand has a first nucleic acid sequence of SEQ ID NO: 23, and the antisense strand has a second nucleic acid sequence of SEQ ID NO:

47. E 32. The SARM1 RNAi agent of any one of E 1 -31 , wherein the sense strand has a delivery moiety conjugated to the 3' end of the sense strand. E 33 . The SARM1 RNAi agent of any one of E 1 -32 , wherein the sense strand has a delivery moiety conjugated to the 5′ end of the sense strand. E 34 . The SARM1 RNAi agent of any one of E 1 -31 , wherein the sense strand has a delivery moiety conjugated to nucleotides of the sense strand.

35. The SARM1 RNAi agent of claim 32 or 33, wherein the delivery moiety is conjugated to the 5' or 3' end of the sense strand via a linker.

36. A pharmaceutical composition comprising the SARM1 RNAi agent of any one of claims 1-35 and a pharmaceutically acceptable carrier.

37. A pharmaceutical composition comprising a means for reducing SARM1 expression in a cell and a pharmaceutically acceptable carrier.

38. A method of reducing SARM1 expression in a patient in need thereof, the method comprising administering to the patient an effective amount of the SARM1 RNAi agent of any one of claims 1-35, or the pharmaceutical composition of claim 36 or 37.

39. A method of reducing axonal degeneration in a patient in need thereof, the method comprising administering to the patient an effective amount of the SARM1 RNAi agent of any one of claims 1-35, or the pharmaceutical composition of claim 36 or 37.

40. A method of treating a SARM1-mediated neurological disease in a patient in need thereof, the method comprising administering to the patient an effective amount of a SARM1 RNAi agent according to any one of claims 1-35, or a pharmaceutical composition according to claim 36 or 37.

41. The method of claim 40, wherein the SARM1-mediated nervous system disease is selected from amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease), Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), Huntington's disease (HD), Alzheimer's disease, Pick's disease, Gaucher's disease, Hurler syndrome, progressive multifocal leukoencephalopathy, Alexander disease, congenital hypomyelination, encephalomyelitis, acute disseminated encephalomyelitis, central pontine myelinolysis, osmotic hyponatremia, Tay-Sachs disease, motor neuron disease, ataxia, spinal muscular atrophy (SMA ), Niemann-Pick disease, acute hemorrhagic leukoencephalitis, trigeminal neuralgia, Bell's palsy, cerebral ischemia, multiple system atrophy, Pelvic floor disease, periventricular leukomalacia, hereditary ataxia, noise-induced hearing loss, congenital hearing loss, age-related hearing loss, Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, dementia with Lewy bodies, frontotemporal dementia, tauopathies, synucleinopathies, amyloidosis, diabetic neuropathy, globoid cell leukodystrophy (Krabbe disease), Baxter-Kohl syndrome, transverse myelitis, motor neuron disease, spinocerebellar ataxia, preeclampsia, hereditary spastic Spastic paraplegia, spastic paraparesis, familial spastic paraplegia, French residency disease, Strumpell-Lorrain disease, nonalcoholic steatohepatitis (NASH), adrenomyoneuropathy, progressive supranuclear palsy (PSP), Friedrich's ataxia, spinal cord injury, acute optic neuropathy (AON), hereditary or idiopathic retinal disorders, Leber's congenital amaurosis (LCA), Leber's hereditary optic neuropathy (LHON), primary open-angle glaucoma (POAG), acute angle-closure glaucoma (AACG), autosomal dominant optic neuropathy atrophy, retinal ganglionic degeneration, retinitis pigmentosa, outer retinal neuropathy, optic neuritis, optic nerve degeneration associated with multiple sclerosis, Kerr's optic neuropathy, ischemic optic neuropathy, chemotherapy-induced peripheral neuropathy, neuromyelitis optica, Charcot-Marie-Tooth disease, vitamin B12 deficiency, folate (vitamin B9) deficiency, isolated vitamin E deficiency syndrome, nonarteritic anterior ischemic optic neuropathy, exposure to ethambutol, exposure to cyanide, traumatic brain injury (TBI), spinal cord injury, traumatic axonal injury, or chronic traumatic encephalopathy (CTE).

42. The method of claim 40 or 41, wherein the SARM1-mediated nervous system disease is amyotrophic lateral sclerosis, multiple sclerosis, chemotherapy-induced peripheral neuropathy (CIPN), diabetic peripheral neuropathy (DPN), Tauopathy, or Charcot-Marie-Tooth disease. E 43. The method of any one of E 40-42, wherein the SARM1 mediated neurological disease is amyotrophic lateral sclerosis. E 44. The method of any one of E 40-43, wherein the SARM1 RNAi agent is administered to the patient intrathecally, intracerebroventricularly, or via intracisternal injection.

45. A method for reducing SARM1 expression in a cell, the method comprising: (a) introducing the SARM1 RNAi agent of any one of claims 1-35 into the cell; as well as (b) incubating the cells for a period of time sufficient to degrade SARM1 mRNA, thereby reducing SARM1 expression in the cells.

46. ​​The SARM1 RNAi agent of any one of claims 1-35 or the pharmaceutical composition of claim 36, for use in therapy.

47. The SARM1 RNAi agent of any one of claims 1-35 or the pharmaceutical composition of claim 36, for use in reducing SARM1 expression.

48. The SARM1 RNAi agent of any one of claims 1-35 or the pharmaceutical composition of claim 36, for use in reducing axonal degeneration.

49. The SARM1 RNAi agent of any one of claims 1-35 or the pharmaceutical composition of claim 36, for use in treating a SARM1-mediated nervous system disease.

50. The SARM1 RNAi agent or pharmaceutical composition for use according to claim 49, wherein the SARM1-mediated nervous system disease is selected from amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease), Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), Huntington's disease (HD), senile dementia, Pick's disease, Gaucher disease, Hurler syndrome, progressive multifocal leukoencephalopathy, Alexander disease, congenital hypomyelination, encephalomyelitis, acute disseminated encephalomyelitis, central pontine myelinolysis, osmotic hyponatremia, Tay-Sachs disease, motor neuron disease, ataxia. disorders, spinal muscular atrophy (SMA), Niemann-Pick disease, acute hemorrhagic leukoencephalitis, trigeminal neuralgia, Bell's palsy, cerebral ischemia, multiple system atrophy, Pelizaeus-Merzheimer disease, periventricular leukomalacia, hereditary ataxia, noise-induced hearing loss, congenital hearing loss, age-related hearing loss, Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, dementia with Lewy bodies, frontotemporal dementia, tauopathies, synucleinopathies, amyloidosis, diabetic neuropathy, globoid cell leukodystrophy (Krabbe disease), Baxter-Kohl syndrome, transverse myelitis, motor neuron disease, spinocerebellar ataxia, Preeclampsia, hereditary spastic paraplegia, spastic paraparesis, familial spastic paraplegia, French residency disease, Strumpell-Lorrain disease, nonalcoholic steatohepatitis (NASH), adrenoleukodystrophy, progressive supranuclear palsy (PSP), Friedrich's ataxia, spinal cord injury, acute optic neuropathy (AON), hereditary or idiopathic retinal disorders, Leber's congenital amaurosis (LCA), Leber's hereditary optic neuropathy (LHON), primary open-angle glaucoma (POAG), acute angle-closure glaucoma (AACG), autosomal dominant optic atrophy, retinal ganglion degeneration, retinitis pigmentosa, outer retinal neuropathy, optic neuritis, optic nerve degeneration associated with multiple sclerosis, Kerr's optic neuropathy, ischemic optic neuropathy, chemotherapy-induced peripheral neuropathy, neuromyelitis optica, Charcot-Marie-Tooth disease, vitamin B12 deficiency, folate (vitamin B9) deficiency, isolated vitamin E deficiency syndrome, nonarteritic anterior ischemic optic neuropathy, exposure to ethambutol, exposure to cyanide, traumatic brain injury (TBI), spinal cord injury, traumatic axonal injury, or chronic traumatic encephalopathy (CTE).

51. The SARM1 RNAi agent or pharmaceutical composition for use according to claim 49 or 50, wherein the SARM1 mediated nervous system disease is amyotrophic lateral sclerosis, multiple sclerosis, chemotherapy-induced peripheral neuropathy (CIPN), diabetic peripheral neuropathy (DPN), Tauopathy or Charcot-Marie-Tooth disease.

52. The SARM1 RNAi agent or pharmaceutical composition for use according to any one of claims 49-51, wherein the SARM1 mediated nervous system disease is amyotrophic lateral sclerosis.

53. Use of the SARM1 RNAi agent of any one of claims 1-35 in the preparation of a medicament for reducing axonal degeneration.

54. Use of the SARM1 RNAi agent of any one of claims 1-35 in the preparation of a medicament for treating a SARM1-mediated nervous system disease.

55. The use of claim 54, wherein the SARM1-mediated nervous system disease is selected from amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease), Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), Huntington's disease (HD), senile dementia, Pick's disease, Gaucher's disease, Hurler syndrome, progressive multifocal leukoencephalopathy, Alexander disease, congenital hypomyelination, encephalomyelitis, acute disseminated encephalomyelitis, central pontine myelinolysis, osmotic hyponatremia, Tay-Sachs disease, motor neuron disease, ataxia, spinal muscular atrophy (SMA ), Niemann-Pick disease, acute hemorrhagic leukoencephalitis, trigeminal neuralgia, Bell's palsy, cerebral ischemia, multiple system atrophy, Pelvic floor disease, periventricular leukomalacia, hereditary ataxia, noise-induced hearing loss, congenital hearing loss, age-related hearing loss, Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, dementia with Lewy bodies, frontotemporal dementia, tauopathies, synucleinopathies, amyloidosis, diabetic neuropathy, globoid cell leukodystrophy (Krabbe disease), Baxter-Kohl syndrome, transverse myelitis, motor neuron disease, spinocerebellar ataxia, preeclampsia, hereditary spastic Spastic paraplegia, spastic paraparesis, familial spastic paraplegia, French residency disease, Strumpell-Lorrain disease, nonalcoholic steatohepatitis (NASH), adrenomyoneuropathy, progressive supranuclear palsy (PSP), Friedrich's ataxia, spinal cord injury, acute optic neuropathy (AON), hereditary or idiopathic retinal disorders, Leber's congenital amaurosis (LCA), Leber's hereditary optic neuropathy (LHON), primary open-angle glaucoma (POAG), acute angle-closure glaucoma (AACG), autosomal dominant optic neuropathy atrophy, retinal ganglionic degeneration, retinitis pigmentosa, outer retinal neuropathy, optic neuritis, optic nerve degeneration associated with multiple sclerosis, Kerr's optic neuropathy, ischemic optic neuropathy, chemotherapy-induced peripheral neuropathy, neuromyelitis optica, Charcot-Marie-Tooth disease, vitamin B12 deficiency, folate (vitamin B9) deficiency, isolated vitamin E deficiency syndrome, nonarteritic anterior ischemic optic neuropathy, exposure to ethambutol, exposure to cyanide, traumatic brain injury (TBI), spinal cord injury, traumatic axonal injury, or chronic traumatic encephalopathy (CTE).

56. The use of claim 54 or 55, wherein the SARM1-mediated nervous system disease is amyotrophic lateral sclerosis, multiple sclerosis, chemotherapy-induced peripheral neuropathy (CIPN), diabetic peripheral neuropathy (DPN), Tauopathy, or Charcot-Marie-Tooth disease.

57. The use of any one of claims 54-56, wherein the SARM1-mediated nervous system disease is amyotrophic lateral sclerosis.

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