CNS targeted SARM1 rnai agents

Abstract

Provided herein are SARM1 RNAi agents and compositions comprising a SARM1 RNAi agent. Also provided herein are methods of using the SARM1 RNAi agents or compositions comprising a SARM1 RNAi agent in reducing SARM1 expression and / or treating SARM1-mediated neurological diseases.

Description

Factors RNAi Addressed to SARM1 In the central nervous system List of sequences The current request is submitted with a sequence list in ST.26 XML format. The sequence list is provided as a file named "30973_WO" created on November 18, 2024, and is 293 KB in size. The ST.26 XML sequence list information is incorporated here by reference in its entirety. Background Axonal degeneration is a pathological feature of numerous neurological diseases, including neurodegenerative diseases, traumatic brain injury, and peripheral neuropathies (Gerdts et al.). Science (Gerdts, J., et al., 2015, 348:453-457). Damaged or unhealthy axons are eliminated through an intrinsic self-destruction program known as erythrodysplasia, an active process of retrograde degeneration of the distal axon terminal while preserving the integrity of the proximal axon portion and cell body (Gerdts, J., et al.). Neuron , 2016, 89, 449-460; Whitmore, A. et al., Cell Death Differ (2003, 10, 260-261). SARM1 (sterile alpha-stimulator and TIR-containing 1) is an NAD+ hydrolase that plays an important role in axonal degeneration. It has been reported that reducing SARM1 expression or eliminating it leads to long-term protection of sensory neurons against injury-induced axonal degeneration (Gerdts et al.). J Neurosci (2013, 33, 13569-13580). WO2021 / 108602 describes an oligonucleotide against the SARM1 trend. The blood-brain barrier (BBB) ​​is a semipermeable, selective boundary in capillary endothelial cells that prevents solutes, including pathogens, from crossing into the central nervous system (CNS). The BBB allows some small molecules to pass through passive diffusion, and BBB cells actively transport metabolic products important for neurological function, such as glucose and amino acids, across the barrier using specific transport proteins. The BBB has a neuroprotective function by tightly controlling access to the brain; however, it also impedes the delivery of therapeutic agents to the CNS. Antibodies targeting the transferrin receptor (TfR) have been used to modulate BBB transport. However, attempts to use TfR antibodies to transport therapeutic agents across the BBB have proven challenging. To date, there are no approved TfR receptor shuttles or conjugates for the treatment of CNS diseases. RNA interference (RNAi) is a highly conserved regulatory mechanism in which RNA molecules are involved in the suppression of a specific sequence of gene expression by double-stranded RNA (dsRNA) molecules (Fire et al., Nature 391:806-811, 1998). There is still a need for therapeutic agents that can inhibit or modulate SARM1 expression to treat SARM1-mediated neurological diseases, for example, through the use of RNAi. Conjugates that can deliver SARM1 RNAi across the blood-brain barrier to the central nervous system are also needed to treat various neurodegenerative diseases of the central nervous system. Invention Summary This document presents SARM1 RNAi factors that can cross the blood-brain barrier and formulations containing SARM1 RNAi. It also presents methods for using SARM1 RNAi factors or formulations containing SARM1 RNAi to reduce SARM1 expression, reduce axonal degeneration, and / or treat SARM1-mediated neurological diseases in a study participant. On one side, SARM1 RNAi factors are provided consisting of the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) consisting of a directional and an antidirectional strand, the antidirectional strand being complementary to SARM1 mRNA; where L is a ligand, or optionally absent; where P is a protein consisting of a monovalent human TfR receptor binding domain ("human TfR receptor binding protein"); and where n is an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3. In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) that includes a directional strand and an anti-directional strand, and the anti-directional strand is complementary to SARM1 mRNA; where L is a linker, or is optionally absent; Where P is a protein consisting of a monovalent human TfR receptor-binding domain, where the human transferrin receptor (TfR)-binding domain includes a heavy chain variant (VH) region and a light chain variant (VL) region, where VH consists of the heavy chain complement mapping regions HCDR1, HCDR2, and HCDR3, while VL consists of the light chain complement mapping regions LCDR1, LCDR2, and LCDR3, where HCDR1 consists of serial identification number: 1, HCDR2 consists of serial identification number: 2, HCDR3 consists of serial identification number: 3, LCDR1 consists of serial identification number: 4, LCDR2 consists of serial identification number: 5, and LCDR3 consists of serial identification number: 6; Where n represents an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3.In some applied models, VH consists of sequence identification number: 7 and VL consists of sequence identification number: 8. In some applied models, VH includes a sequence that matches at least 95% with sequence identification number: 7, and VL includes a sequence that matches at least 95% with sequence identification number: 8. Typical sequences of human transferrin receptor binding domains and proteins are provided in Tables 1A and 1B. In some implementation models, L is a Mal-Tet-TCO link, an SMCC link, a GDM link, an MSPT link, or an OD link (see Table 4). In some implementation models, L is an SMCC link in Table 4. In some implementation models, L is an MSPT link in Table 4. Table 5 provides examples of unmodified directional and anti-directional strand sequences of dsRNA targeting human SARM1 mRNA. In some applied models, the directional and anti-directional strands of dsRNA include a pair of DNA sequences selected from a set that includes the following: (a) The directional braid consists of the serial identification number: 35, while the anti-directional braid consists of the serial identification number: 36; (b) The directional braid consists of the serial identification number: 37, while the anti-directional braid consists of the serial identification number: 38; (c) The directional braid consists of the serial identification number: 39, while the anti-directional braid consists of the serial identification number: 40; and (d) The directional braid consists of the serial identification number: 41, while the anti-directional braid consists of the serial identification number: 42; Where one or more of the nucleotides in the directional and anti-directional strands are independently modified nucleotides, and where one or more of the nucleotide-linking bonds in the directional and anti-directional strands are independently modified nucleotide-linking bonds. In some applied models, the directional strand consists of serial identification number 35, while the anti-directional strand consists of serial identification number 36. In some applied models, the directional strand consists of serial identification number 37, while the anti-directional strand consists of serial identification number 38. In some applied models, the directional strand consists of serial identification number 39, while the anti-directional strand consists of serial identification number 40. In some applied models, the directional strand consists of serial identification number 41, while the anti-directional strand consists of serial identification number 42. dsRNA can undergo modifications. These modifications can be made to one or more of the oriented and / or anti-oriented strand nucleotides, or to the bonds between nucleotides. In some applied models, one or more of the oriented and / or anti-oriented strand nucleotides are independently modified nucleotides, meaning that the oriented and anti-oriented strands can contain different modified nucleotides. In some applied models, each nucleotide of the oriented strand is a modified nucleotide. In some applied models, each nucleotide of the anti-oriented strand is a modified nucleotide. In some applied models, the modified nucleotide is a 2'-fluoro-modified nucleotide, a 2'-methyl-modified nucleotide, a 2'-deoxynucleotide (DNA), or a 2'-alkyl-modified nucleotide (e.g., alkyl 2'-OC). 16 In some applied models, each nucleotide of the directional and antidirectional strands is an independently modified nucleotide, for example, a fluoro-2'-modified nucleotide, a methyl-2'-O-modified nucleotide, a 2'-deoxynucleotide (DNA), or a 2'-O-modified alkyl nucleotide (e.g., alkyl 2'-OC). 16 ). In some applied models, the directional strand includes four fluoro-2' modified nucleotides at positions 7, 9, 10, and 11 of the 5' end of the directional strand. In some applied models, at least one nucleotide of the directional strand is an unmodified RNA nucleotide. In some applied models, at least one nucleotide of the directional strand is a 2' deoxyribonucleotide (DNA). In some applied models, the other nucleotides of the directional strand are 2'-O-methyl modified nucleotides. In some applied models, the counter-directional strand includes four fluoro-2' modified nucleotides, for example, at positions 2, 6, 14, and 16 of the 5' end of the counter-directional strand. In some applied models, the other nucleotides of the counter-directional strand are 2'-O-methyl modified nucleotides. In some applied models, the directional strand contains three fluoro-2' modified nucleotides at positions 9, 10, and 11 of the 5' end of the directional strand. In some applied models, at least one nucleotide of the directional strand is an unmodified RNA nucleotide. In some applied models, at least one nucleotide of the directional strand is a 2' deoxyribonucleotide (DNA). In some applied models, the remaining nucleotides of the directional strand are 2'-O-methyl modified nucleotides. In some applied models, the counter-directional strand contains five fluoro-2' modified nucleotides, for example, at positions 2, 5, 7, 14, and 16 of the 5' end of the counter-directional strand. In some applied models, the anti-directional strand includes five fluoro-2' modified nucleotides, for example, at positions 2, 5, 8, 14, and 16 of the 5' end of the anti-directional strand.In some applied models, the antiderivative strand includes three 2'-fluoro-modified nucleotides, for example, at positions 2, 14, and 16 of the 5' end of the antiderivative strand. In other applied models, the other nucleotides of the antiderivative strand are 2'-O-methyl-modified nucleotides. In some applied models, the 5' end of the strand against the direction contains a phosphate isotope, for example 5'-phenylphosphonate (5'-VP). In some applied models, the trend braid or anti-trend braid includes a non-stepping portion or an inverse non-stepping portion. In some applied models, the trending and counter-trend strands contain one or more bonds between modified nucleotides. In some applied models, the bond between modified nucleotides is a phosphorothiolate bond. In some applied models, the trending strand contains four or five phosphorothiolate bonds. In some applied models, the counter-trend strand contains four or five phosphorothiolate bonds. In some applied models, both the trending and counter-trend strands contain four phosphorothiolate bonds. And in some applied models, the trending strand contains four phosphorothiolate bonds and the counter-trend strand contains five phosphorothiolate bonds. Table 7 provides examples of modified directional and anti-directional strand sequences of dsRNA targeting human SARM1 mRNA. In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) consisting of a directional and an antidirectional strand, and the antidirectional strand is complementary to SARM1 mRNA; where L is a linker, or optionally absent; where P is a protein consisting of a single monovalent TfR receptor binding domain, and P is chosen from TBP1, TBP2, TBP3, TBP4, or TBP5 in Table 1b; and where n is an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3. In some applied models, L is a linker in Table 4 (for example, the SMCC or MSPT linker in Table 4). In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) that includes a directional strand and an antidirectional strand, and dsRNA is any dsRNA in Table 5 or 7; where L is a linker, or optionally absent; where P is a protein consisting of a monovalent human TfR binding domain; and where n is an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3. In some applied models, L is a linker in Table 4 (for example, the SMCC or MSPT linker in Table 4). In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) that includes a directional and an antidirectional strand, and dsRNA is any dsRNA in Table 5 or 7; where L is a linker, or optionally absent; where P is a protein consisting of a single monovalent TfR receptor binding domain, and P is chosen from TBP1, TBP2, TBP3, TBP4, or TBP5 in Table 1b; and where n is an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3. In some applied models, L is a linker in Table 4 (for example, the SMCC or MSPT linker in Table 4). In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) consisting of a directional and an antidirectional strand, and the antidirectional strand is complementary to SARM1 mRNA; where L is a linker, or is optionally absent; where P is a protein consisting of a monovalent human TfR binding domain; the human TfR receptor binding domain consisting of a pair of heavy chains HC1 and HC2 and one light chain LC1, where HC1 consists of the sequence ID: 14, LC1 consists of the sequence ID: 10, and HC2 consists of the sequence ID: 15, where n is equal to 1 or 2. In some applied models, n is 1. In some applied models, n is 2. In some applied models, L is a linker in Table 4 (for example, the SMCC or MSPT linker in Table 4). In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) that includes a directional strand and an antidirectional strand, and the antidirectional strand is complementary to SARM1 mRNA; where L is a linker, or is optionally absent; where P is a protein consisting of a monovalent human TfR binding domain; where the human TfR receptor binding domain consists of a pair of heavy chains HC1 and HC2 and one light chain LC1, where HC1 consists of sequence ID: 16, LC1 consists of sequence ID: 10, and HC2 consists of sequence ID: 17, where n is equal to 1. In some applied models, L is a linker in Table 4 (e.g., SMCC or MSPT linker in Table 4). On the other hand, there are methods available here for treating SARM1-mediated neurological disease in patients who require it. This involves administering an effective dose of SARM1 RNAi or a pharmaceutical formulation described here. SARM1 RNAi or a pharmaceutical formulation containing SARM1 RNAi can be administered intravenously or subcutaneously. On the other hand, this document provides information on SARM1 RNAi agents or pharmaceutical formulations containing SARM1 RNAi for therapeutic use. It also provides information on SARM1 RNAi agents or pharmaceutical formulations containing SARM1 RNAi for use in the treatment of SARM1-mediated neurological disease. Furthermore, it details the applications of SARM1 RNAi in the manufacture of a drug for the treatment of SARM1-mediated neurological disease. Brief description of the drawings It is clear Figure 1a A typical preparative anion-exchange chromatogram (AEX) of the drug-antibody ratio (DAR) pattern of TBP5-SMCC-dsRNA conjugate #8 before purification. It illustrates Figure 1b A typical anion-exchange chromatogram for the antibody-drug ratio (DAR) pattern of TBP5-SMCC-dsRNA conjugate #8 after purification. It illustrates Figure 1c A typical anion-exchange chromatogram (AEX) of the drug-antibody ratio (DAR) pattern of TBP5-SMCC-dsRNA conjugate #7 before purification. It illustrates Figure 1d A typical anion-exchange chromatogram (AEX) of the drug-antibody ratio (DAR) pattern of TBP5-SMCC-dsRNA conjugate #7 after purification. It illustrates Figure 1E A typical anion-exchange chromatogram (AEX) of the drug-antibody ratio (DAR) pattern of TBP5-SMCC-dsRNA conjugate #6 before purification. It illustrates Figure 1 A typical analytical anion exchange chromatogram for the antibody-drug ratio (DAR) pattern of TBP5-SMCC-dsRNA conjugate No. 6 after purification. It is clear Figure 2 SARM1 protein reduction In the laboratory In SH-SY5Y cells using unconjugated dsRNA No. 8, cholesterol-conjugated dsRNA No. 8, and TBP5-conjugated dsRNA No. 8. It is clear Figures 3a and 3b Proof-of-concept data in hTfR mice, demonstrating the pharmacodynamic efficacy of TBP5-SARM1 dsRNA conjugate #8 when administered either as a single intravenous dose of 10 mg / kg siRNA or four weekly intravenous doses of 10 mg / kg. A decrease in SARM1 mRNA in the mouse brain hemisphere (3A) and gastrocnemius muscle (3B) is shown 28 days after the first dose. Both dose groups showed a significant decrease in brain SARM1 mRNA compared to the untreated control groups (PBS group). Error bars represent standard deviations. Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparisons test against the phosphate-buffered saline control group. Annotations indicate that possible values ​​are: >0.0001=****; >0.001=***; >0.01=**; >0.05=*. He explains Figure 4A A decrease in SARM1 protein levels was observed in rhesus monkey tissues following a single intravenous dose of TBP5-dsRNA conjugate number 7 at 10 mg / kg. Figure 4b SARM1 protein levels decreased in rhesus monkey tissues after administration of a single intravenous peripheral dose of TBP5-dsRNA conjugate No. 6 at 10 mg / kg of siRNA. He explains Figure 5 A typical analytical anion exchange chromatogram for the antibody-drug ratio (DAR) pattern of TBP5-MSPT-dsRNA conjugate No. 8 after purification. It is clear Figures 6a and 6b Data from hTfR transgenic mice, demonstrating the pharmacodynamic efficacy of TBP5-SMCC-dsRNA No. 8 or TBP5-MSPT-dsRNA No. 8 conjugates when administered as a single intravenous dose of 10 mg / kg siRNA. A decrease in SARM1 mRNA is shown in the mouse brain hemisphere (6A) and lumbar spinal cord (6B) 28 days after the first dose. Both dose groups showed a statistically significant decrease in SARM1 mRNA in the brain and spinal cord compared to the PBS-treated control groups. Error bars represent standard deviations. Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparisons test against the phosphate-buffered saline control group. The annotations indicate that possible values ​​are: >0.0001=****; >0.001=***; >0.01=**; >0.05=*. Detailed description of the invention This document provides information on SARM1 RNAi factors and formulations containing SARM1 RNAi. It also presents methods for using SARM1 RNAi factors or formulations containing SARM1 RNAi to reduce SARM1 expression, reduce axonal degeneration, and / or treat SARM1-mediated neurological diseases in a study participant. On one side, SARM1 RNAi factors are provided consisting of the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) consisting of a directional and an antidirectional strand, where the antidirectional strand is complementary to SARM1 mRNA; where L is a ligand, or optionally absent; where P is a protein consisting of a monovalent human TfR receptor binding domain ("human TfR receptor binding protein"); and where n is an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3. In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) that includes a directional strand and an antidirectional strand, and the antidirectional strand is complementary to SARM1 mRNA; where L is a linker, or optionally absent; where P is a protein consisting of a monovalent human TfR binding domain; The human transferrin receptor (TfR) binding domain comprises a heavy-chain variable (VH) region and a light-chain variable (VL) region, where VH consists of the heavy-chain complement mapping regions HCDR1, HCDR2, and HCDR3, and VL consists of the light-chain complement mapping regions LCDR1, LCDR2, and LCDR3, where HCDR1 has serial identification number 1, HCDR2 has serial identification number 2, HCDR3 has serial identification number 3, LCDR1 has serial identification number 4, LCDR2 has serial identification number 5, and LCDR3 has serial identification number 6; and where n represents an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3.In some applied models, L is a link in Table 4 (for example, the SMCC or MSPT link in Table 4). In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) consisting of a directional and an antidirectional strand, and the antidirectional strand is complementary to SARM1 mRNA; where L is a linker, or optionally absent; where P is a protein consisting of a single monovalent TfR receptor binding domain, and P is chosen from TBP1, TBP2, TBP3, TBP4, or TBP5 in Table 1b; and where n is an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3. In some applied models, L is a linker in Table 4 (for example, the SMCC or MSPT linker in Table 4). In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) that includes a directional and an antidirectional strand, and dsRNA is any dsRNA in Table 5 or 7 (e.g., dsRNA No. 1); where L is a linker, or is optionally absent; where P is a protein consisting of a monovalent human TfR binding domain; and where n is an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3. In some applied models, L is a linker in Table 4 (e.g., the SMCC or MSPT linker in Table 4). In some applied models, this document provides SARM1 RNAi factors that include the formula (I): (RL) n -P, where R is a double-stranded RNA (dsRNA) that includes a directional strand and an antidirectional strand, and dsRNA is any dsRNA in Table 5 or 7 (e.g., dsRNA No. 1); where L is a linker, or is optionally absent; where P is a protein consisting of a single monovalent TfR receptor binding domain, and P is chosen from TBP1, TBP2, TBP3, TBP4, or TBP5 in Table 1b; and where n is an integer from 1 to 3. In some applied models, n is 1. In some applied models, n is 2. In some applied models, n is 3. In some applied models, L is a linker in Table 4 (e.g., SMCC or MSPT linker in Table 4). receptor-binding proteins TfR human The SARM1 RNAi factors described in this document consist of a protein containing a single monovalent human TfR receptor-binding domain (“human TfR-binding protein”). The human TfR-binding protein in SARM1 RNAi factors can bind to the TfR receptor on the blood-brain barrier and deliver dsRNA to the central nervous system. Typical sequences of human transferrin receptor binding domains and proteins are provided in Tables 1A and 1B. In some applied models, the human monovalent TfR receptor binding domain includes a heavy chain variant (VH) region and a light chain variant (VL) region. The VH region includes the heavy chain complement mapping regions HCDR1, HCDR2, and HCDR3, and the VL region includes the light chain complement mapping regions LCDR1, LCDR2, and LCR3. In some application models, the HCDR1 region consists of serial identification number: 1, the HCDR2 region consists of serial identification number: 2, the HCDR3 region consists of serial identification number: 3, the LCDR1 region consists of serial identification number: 4, the LCDR2 region consists of serial identification number: 5, and the LCDR3 region consists of serial identification number: 6. In some application models, VH consists of serial identification number: 7 and VL consists of serial identification number: 8.In some applied models, VH includes a sequence that matches at least 95% with serial identification number: 7, and VL includes a sequence that matches at least 95% with serial identification number: 8. Table 1A. Typical sequences of proteins and receptor-binding domains TfR human area Sequence Serial identification number HCDR1 (KABAT) SYSMN 1 HCDR2 (KABAT) SISSSSSYIYYADSVKG 2 HCDR3 (KABAT) RHGYSNSDAFDN 3 LCDR1 (KABAT) RASQGISHYLV 4 LCDR2 (KABAT) AASSLQS 5 LCDR3 (KABAT) LQHNSYPWT 6 VH EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSS 7 VL DIQMTQSPSAMSASVGDRVTITCRASQGISHYLVWFQQKPGKVPKRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPWTFGQGTKVEIK 8 Fab HC EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKC 9 Fab LC / Fab-VHH LC DIQMTQSPSAMSASVGDRVTITCRASQGISHYLVWFQQKPGKVPKRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 10 Fab-VHH HC EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSSASTKGPCVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKCDKTHTGGGGQGGGGQGGGGQGGGGQGGGGQEVQLLESGGGLVQPGGSLRLSCAASGRYIDETAVAWFRQAPGKGREFVAGIGGGVDITYYADSVKGRFTISRDNSKNTLYLQMNSLRPEDTAVYYCGARPGRPLITSKVADLYPYWGQGTLVTVSSPP 11 Fab-VHH LC DIQMTQSPSAMSASVGDRVTITCRASQGISHYLVWFQQKPGKVPKRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQCGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 12 hIgG4 PAA HC EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSSASTKGPXVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG، حيث تكون X عبارة عن S أو C. 13 OAH1 (هيتروماب أحادي الذراع) HC1 (A378C) EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVSTLPPSQEEMTKNQVSLMCLVYGFYPSDIXVEWESNGQPENNYKTTPPVLDSDGSFFLYSVLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG، حيث يكون X عبارة عن A أو C. 14 OAH1 HC2 (A378C) ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQGDMTKNQVQLTCLVKGFYPSDIXVEWESNGQPENNYKTTPPVLDSDGSFFLASRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG، حيث يكون X عبارة عن A أو C. 15 OAH LC DIQMTQSPSAMSASVGDRVTITCRASQGISHYLVWFQQKPGKVPKRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 10 OAH2 HC1 (S124C) EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSSASTKGPCVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVSTLPPSQEEMTKNQVSLMCLVYGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSVLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 16 OAH2 HC2 (S124C) ESKYGPPCPPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQGDMTKNQVQLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLASRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 17 HC For example Zero QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSYAIEWVRQAPGQGLEWMGGILPGSGTINYNEKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARMSSNSDQGFDLWG QGTLVTVSSASTKGPXVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPEEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG, Where X is S, or C. 18 LC arm Null DIQMTQSPSSLSSASVGDRVTITCKASQGISRFLSWFQQKPGKAPKSLIYAVSSLVDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCVQYNSYPYGFGGGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 19 Table 1b. Typical sequences of receptor-binding proteins TfR human receptor binding protein TfR human ( TBP) HC1 LC1 HC2 LC2 TBP1 (Fab) Serial identification number: 9 Serial ID number: 10 unavailable unavailable TBP2 (Fab-VHH) Serial ID number: 11 Serial identification number: 12 or 10 unavailable unavailable TBP3 (Heterogeneous dimeric antibody) Serial identification number: 13 Serial ID number: 10 Serial ID number: 18 Serial ID number: 19 TBP4 (Hytromap Single Arm 1, A378C) Serial ID number: 14 Serial ID number: 10 Serial ID number: 15 N / A* TBP5 (Hytromap Single Arm 2, S124C) Serial ID number: 16 Serial ID number: 10 Serial ID number: 17 N / A* In some applied models, the monovalent human TfR receptor binding domain is a subunit of the antibody, e.g., Fab, scFv, Fv, or scFab (the single-chain Fab subunit). In other applied models, the monovalent human TfR receptor binding domain is the Fab subunit. And in still other applied models, the human TfR receptor binding domain also consists of a heavy-chain constant region and / or a light-chain constant region. In some applied models, the human TfR receptor-binding protein also includes a half-life extender, for example, the Fc region of immunoglobulin or VHH that binds human serum albumin (HSA). In some applied models, the human TfR receptor-binding protein also includes an immunoglobulin Fc region, e.g., the Fc region of modified human IgG4, or the Fc region of modified human IgG1. In some applied models, the human TfR receptor-binding protein also includes an Fc region of modified human IgG4 consisting of proline in residue 228, and alanine in residues 234 and 235 (all residues are numbered according to the EU index numbering, also called the hIgG4PAA Fc region). In some applied models, the human TfR receptor-binding protein also includes a modified human IgG1 Fc region consisting of alanine at residues 234, 235, and 329, serine at position 265, and aspartic acid at position 436 (all residues are numbered according to the EU index numbering, also called the non-responder hIgG1 or the hIgG1EN Fc region). In some applied models, the human TfR receptor-binding protein also includes a VHH that binds to human serum albumin. In some applied models, the VHH also binds to albumin from mice, rats, and / or long-tailed macaques. Table 2 shows a model of a VHH that binds to human serum albumin. In some applied models, the VHH consists of CDR1 (serial ID: 20), CDR2 (serial ID: 21), and CDR3 (serial ID: 22). In some applied models, the VHH includes the serial ID: 23. In some applied models, the VHH is linked to the TfR receptor-binding domain via a peptide linker, for example, (GGGGQ)4 (serial ID: 24). In some applied models, the VHH is linked to the C-terminus of the TfR receptor-binding domain. Table 2. Typical sequences for VHH Which is related to human serum albumin ( (HSA) area Sequence Serial identification number CDR1 (KABAT) ETAVA 20 CDR2 (KABAT) GIGGGVDITYYADSVKG 21 CDR3 (KABAT) RPGRPLITSKVADLYPY 22 The full length of VHH EVQLLESGGGLVQPGGSLRLSCAASGRYIDETAVAWFRQAPGKGREFVAGIGGGVDITYYADSVKGRFTISRDNSKNTLYLQMNSLRPEDTAVYYCGARPGRPLITSKVADLYPYWGQGTLVTVSSPP 23 الرابط الاختياري GGGGQGGGGQGGGGQGGGGQ 24 In some applied models, the human TfR receptor-binding protein is a heterodimeric antibody consisting of a first arm containing a single binding domain to the monovalent human TfR receptor and a second arm that is empty, for example, an arm that does not bind to any known human target (e.g., the isotype arm). Heteromab, orthomab, and dopodi-type heterodimeric antibodies have been described in WO2014150973, WO2016118742, WO2018118616, and WO2011131746. In some applied models, the first arm contains any of the monovalent human TfR receptor-binding domains described in this document. In some applied models, the second arm is empty and does not bind to any known human target (e.g., the isotype arm) and consists of the sequences described in Table 1A. In some applied models, the second arm includes a heavy chain (HC) and a light chain (LC), where the HC consists of serial identification number: 18, and the LC consists of serial identification number: 19. In some applied models, the human TfR receptor-binding protein includes heterodimeric mutations. In some applied models, the human TfR receptor-binding protein described in this document includes a modified Fc region consisting of an Fc CH3 1 domain containing serine at residue 349, methionine at residue 366, tyrosine at residue 370, and valine at residue 409, and an Fc CH3 2 domain containing glycine at residue 356, aspartic acid at residue 357, glutamine at residue 364, and alanine at residue 407 (all residues are numbered according to the EU index numbering). In some applied models, the human TfR receptor-binding protein described in this document includes a modified Fc region consisting of an Fc CH3 1 domain containing leucine at residue 405, and an Fc CH3 2 domain containing arginine at residue 409 (all residues are numbered according to the EU index numbering). In some applied models, the human TfR receptor-binding protein contains one or more native cysteine ​​residues, which can be used for conjugation. For example, in some applied models, the human TfR receptor-binding protein contains a native cysteine ​​at position 220 of the light chain and / or a native cysteine ​​at position 226 of the heavy chain, which can be used for conjugation (all residues are numbered according to the EU index numbering). In some applied models, the human TfR receptor-binding protein (TRB) includes engineered cysteine ​​residues for conjugation. An approach to including engineered cysteines as a conjugation method has been described in WO 2018 / 232088. In some applied models, the TRB includes a heavy chain consisting of one or more cysteines in the following residues: 124, 157, 162, 262, 373, 375, 378, 397, and 415 (all residues are numbered according to EU index numbering). In some applied models, the TRB includes a light chain (e.g., kappa light chain) consisting of one or more cysteines in the following residues: 156, 171, 191, 193, 202, and 208 (all residues are numbered according to EU index numbering). In some applied models, the human TfR receptor-binding protein includes a stable heavy chain region containing cysteine ​​at residual 124 (according to EU index numbering).In some applied models, the human TfR receptor-binding protein includes a stable light-chain region containing cysteine ​​at residual 156 (according to EU index numbering). In other applied models, the human TfR receptor-binding protein includes an immunoglobulin Fc region composed of cysteine ​​at residual 378 (according to EU index numbering). In some applied models, the human TfR receptor-binding protein is any of the human TfR receptor-binding proteins in Table 1B, for example, TBP1, TBP2, TBP3, TBP4, and TBP5. In some applied models, the human TfR receptor-binding protein has a Fab format, for example, TBP1. In some applied models, the human TfR receptor-binding protein consists of HC and LC, where HC consists of sequence identification number: 9, and LC consists of sequence identification number: 10. In some applied models, the human TfR receptor-binding protein has the formula Fab-VHH, for example, TBP2. In some applied models, the human TfR receptor-binding protein consists of HC and LC; where HC consists of serial identification number: 11, and LC consists of serial identification number: 12 or 10. In some applied models, the human TfR receptor-binding protein contains a heterodimeric antibody formulation, for example, TBP3. In some applied models, the human TfR receptor-binding protein consists of a pair of heavy chains HC1 and HC2 and a pair of light chains LC1 and LC2, where HC1 consists of serial identification number: 13, LC1 consists of serial identification number: 10, while HC2 consists of serial identification number: 18, and LC2 consists of serial identification number: 19. In some applied models, the human TfR receptor-binding protein has a single-arm hetromab conformation, for example, TBP4 or TBP5. In some applied models, the human TfR receptor-binding protein consists of a pair of heavy chains HC1 and HC2 and a single light chain LC1, where HC1 has serial identification number 14, LC1 has serial identification number 10, and HC2 has serial identification number 15. In some applied models, human TfR receptor-binding proteins consisting of a pair of heavy chains HC1 and HC2 and a single light chain LC1 are available in this document, where HC1 has serial identification number 16, LC1 has serial identification number 10, and HC2 has serial identification number 17. The human TfR receptor-binding proteins described in this document can be produced in a recombinant manner in a host cell, for example, using an expression vector. For instance, an expression vector might contain a sequence encoding one or more signaling peptides that facilitate the secretion of a polypeptide (polypeptide) from a host cell. Expression vectors containing a polynucleotide of interest (e.g., a polynucleotide encoding a heavy or light chain of a TfR receptor-binding protein) can be transferred into a host cell using known methods. Additionally, expression vectors might contain one or more selection markers, such as tetracycline, neomycin, and dihydrofolate reductase, to aid in the detection of mutant host cells with the desired polynucleotide sequences.The host cell contains stably or transiently transfected, transformed, or transduced cells, or cells infected with one or more expression vectors expressing all or some of the TfR receptor-binding proteins described in this document. In some applied models, the host cell may be stably or transiently transfected, transformed, or infected with an expression vector expressing both heavy-chain polypeptides and light-chain polypeptides of the TfR receptor-binding proteins described in this document. In other applied models, the host cell may be stably or transiently transfected, transformed, or infected with an expression vector expressing both heavy-chain and light-chain polypeptides of the TfR receptor-binding proteins described in this document. TfR receptor-binding proteins can be produced in mammalian cells such as CHO, NS0, HEK293, or COS cells using established techniques. The medium in which TfR receptor-binding proteins were secreted can be purified using conventional techniques, such as mixed-mode ion-exchange chromatography and hydrophobic reaction chromatography. For example, the medium can be placed on a protein A or G column and the protein extracted using conventional methods; mixed-mode ion-exchange chromatography and hydrophobic reaction chromatography can also be employed. Soluble aggregations and polymers can be effectively removed by common techniques, including size exclusion, hydrophobic reaction, ion-exchange, and hydroxyapatite chromatography. Various methods can be used for protein purification, and these methods are well-known in the field and described, for example, in Deutscher, Methods in Enzymology 182: 83-89 (1990) and Scopes, Protein Purification: Principles and Practice, 3rd ed., Springer, NY (1994). receptor-binding proteins TfR mouse Some of the SARM1 RNAi factors used in the examples below include a protein consisting of a mouse monovalent TfR receptor-binding domain (“mouse TfR receptor-binding proteins” or mTBP). Typical sequences of mouse TfR receptor-binding proteins are provided in Table 3. SARM1 RNAi factors containing the mouse TfR receptor-binding protein can serve as alternative molecules in mouse models to SARM1 RNAi factors containing the human TfR receptor-binding protein. Table 3. Typical sequences of receptor-binding protein TfR The mouse ( mTBP1) area Sequence Serial identification number HCDR1 (KABAT) GSYWIC 25 HCDR2 (KABAT) CIYSTSGGRTYYASWVKG 26 HCDR3 (KABAT) GDDSISDAYFDL 27 LCDR1 (KABAT) QSSQSVYNNNRLA 28 LCDR2 (KABAT) DASTLAS 29 LCDR3 (KABAT) QGTYFSSGWSWA 30 VH QSLEESGGDLVKPEGSLTLTCTASGFSFSGSYWICWVRQAPGKGLEWIGCIYSTSSGGRTYYASWVKGRFTISKTSSTTVTLQMTSLTAADTATYFCARGDDSISDAYFDLWGPGTLVTVSS 31 VL ALDMTQTASPVSAAVGGTVTINCQSSQSVYNNNRLAWYQQKPGQPPKLLIYDASTLASGVPSRFKGSGSGTQFTLTISGVQSDDSATYYCQGTYFSSGWSWAFGGGTEVVVK 32 HC1 QSLEESGGDLVKPEGSLTLTCTASGFSFSGSYWICWVRQAPGKGLEWIGCIYSTSGGRTYYASWVKGRFTISKTSSTTVTLQMTSLTAADTATYFCARGDDSISDAYFDLWGPGTLVTVSSASTKGPCVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 33 LC1 ALDMTQTASPVSAAVGGTVTINCQSSQSVYNNNRLAWYQQKPGQPPKLLIYDASTLASGVPSRFKGSGSGTQFTLTISGVQSDDSATYYCQGTYFSSGWSWAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 34 HC لذراع Null ‏( HC2) QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSYAIEWVRQAPGQGLEWMGGILPGSGTINYNEKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARMSSSNSDQGFDLWGQGTLVTVSSASTKGPXVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPPAPEAAGGPSVFLFPPPKPKDTLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG, where X is an S or a C. 18 LC For example Zero ‏( LC2) DIQMTQSPSSLSASVGDRVTITCKASQGISRFLSWFQQKPGKAPKSLIYAVSSLVDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCVQYNSYFGGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 19 The link In some applied models, the SARM1 RNAi factors described in this document include a linker that connects the human TfR receptor-binding protein to dsRNA. In some applied models, the linker is a Mal-Tet-TCO linker, an SMCC linker, a GDM linker, an MSPT linker, or an OD linker (Table 4 shows the structures of these linkers). In some applied models, the linker is an SMCC linker. In some applied models, the linker is an MSPT linker. Table 4. Typical bond combinations Link Structure** 1 SMCC Link 1* 2 SMCC Link 2 3 The open shape of the hydrolyzed ring of the SMCC 1* linker 4 The open-loop hydrolyzed structure of the SMCC 2 linker 5 Mal-Tet-TCO 1* link 6 Mal-Tet-TCO 2 link 7 GDM Link 1* 8 GDM linker2 9 Link 3'-OD* 10 Link 3'-MSPT* 11 Link 5'-OD 12 Link 5'-MSPT *Note - X is either O or S **It is understood by experts in the field that the sulfur bond associated with TBP can be considered part of TBP. dsRNA The SARM1 RNAi factors described in this document consist of double-stranded RNA (dsRNA) containing a directional and an antidirectional strand, where the antidirectional strand is complementary to SARM1 mRNA. After incorporating the antidirectional strand of the dsRNA into the resulting RNA quenching complex (RISC), the RISC complex can bind to and degrade the target SARM1 mRNA. In some applied models, the length of both the directional and antidirectional strands of dsRNA is estimated to be 15–30 nucleotides, for example, 20–25 nucleotides. In other applied models, dsRNA consists of a directional strand of 21 nucleotides and an antidirectional strand of 23 nucleotides. In some applied models, the directional and antidirectional strands of dsRNA may have buoyancy at the 5' or 3' ends (i.e., a 5' buoyancy or a 3' buoyancy). For example, the directional and antidirectional strands may contain a 5' or 3' buoyancy of 1 to 5 nucleotides or 1 to 3 nucleotides. In some applied models, the antidirectional strand includes a 3' buoyancy of two nucleotides. Table 5 provides examples of unmodified directional and anti-directional strand sequences of dsRNA targeting human SARM1 mRNA. Table 5. Unmodified sequences from dsRNA Which targets SARM1 mRNA human number dsRNA Directional braid ( 5' to 3' ) Serial identification number The braid is against the direction ( 5' to 3' ) Serial identification number The starting position of the target area for the braid is against the direction of the copy. SARM1 Humanity NM_015077.4* 1 GUUGCUCGACUCUAACCGCUA 35 UAGCGGUUAGAGUCGAGCAACGG 36 1333 2 UUCGCCAACUAUUCUACGUGA 37 UCACGUAGAAUAGUUGGCGAAGG 38 1763 3 ACCUUCGCCAACUAUUCUACA 39 UGUAGAAUAGUUGGCGAAGGUCU 40 1760 4 CCGCAAGAGGUUCUUUAGGGA 41 UCCCUAAAGAACCUCUUGCGGGU 42 1723 In some applied models, the directional and anti-directional strands in dsRNA involve a pair of DNA sequences selected from a set that includes the following: (a) The directional braid consists of the serial identification number: 35, while the anti-directional braid consists of the serial identification number: 36; (b) The directional braid consists of the serial identification number: 37, while the anti-directional braid consists of the serial identification number: 38; (c) The directional braid consists of the serial identification number: 39, while the anti-directional braid consists of the serial identification number: 40; and (d) The directional braid consists of the serial identification number: 41, while the anti-directional braid consists of the serial identification number: 42; Where one or more of the nucleotides in the directional and anti-directional strands are independently modified nucleotides, and where one or more of the nucleotide-linking bonds in the directional and anti-directional strands are independently modified nucleotide-linking bonds. In some applied models, the directional strand consists of serial identification number 35, while the anti-directional strand consists of serial identification number 36. In some applied models, the directional strand consists of serial identification number 37, while the anti-directional strand consists of serial identification number 38. In some applied models, the directional strand consists of serial identification number 39, while the anti-directional strand consists of serial identification number 40. In some applied models, the directional strand consists of serial identification number 41, while the anti-directional strand consists of serial identification number 42. dsRNA can undergo modifications. These modifications can be made to one or more strand-directing and / or anti-directing nucleotides, or to internucleotide bonds, which are the bonds between two nucleotides in the strand-directing or anti-directing strand. For example, certain modifications at the 2' position of ribose or deoxyribose can increase the stability and half-life of RNA or DNA. These 2' modifications can be 2'-fluoro, 2'-O-methyl (i.e., 2'-methoxy), or 2'-O-alkyl (e.g., alkyl 2'-OC). 16 ). In some applied models, one or more of the oriented strand and / or anti-oriented strand nucleotides are independently modified nucleotides, meaning that the oriented strand and anti-oriented strand can contain different modified nucleotides. In some applied models, each nucleotide of the oriented strand is a modified nucleotide. In some applied models, at least one nucleotide of the oriented strand is an unmodified RNA nucleotide. In some applied models, each nucleotide of the anti-oriented strand is a modified nucleotide. In some applied models, the modified nucleotide is a 2'-fluoro-modified nucleotide, a 2'-methyl-O-modified nucleotide, a 2'-deoxynucleotide (DNA), or a 2'-Alkyl-O-modified nucleotide (e.g., alkyl 2'-OC). 16 In some applied models, each nucleotide of the directional and antidirectional strands is an independently modified nucleotide, for example, a fluoro-2'-modified nucleotide, a methyl-2'-O-modified nucleotide, a 2'-deoxynucleotide (DNA), or a 2'-O-modified alkyl nucleotide (e.g., alkyl 2'-OC). 16 In some applied models, at least one nucleotide of the directional strand is a 2' deoxynucleotide (DNA). In some applied models, the directional strand includes four 2'-fluoro-modified nucleotides at positions 7, 9, 10, and 11 of the 5' end of the directional strand. In some applied models, at least one of the directional strand nucleotides is an unmodified RNA nucleotide. In some applied models, at least one of the directional strand nucleotides is a 2'-deoxynucleotide (DNA). In some applied models, the remaining nucleotides of the directional strand are 2'-O-methyl-modified nucleotides. In some applied models, the antiderivative strand includes four 2'-fluoro-modified nucleotides, for example, at positions 2, 6, 14, and 16 of the 5' end of the antiderivative strand. In other applied models, the remaining nucleotides of the antiderivative strand are 2'-O-methyl-modified nucleotides. In some applied models, the directional strand includes three 2'-fluoro-modified nucleotides at positions 9, 10, and 11 of the 5' end of the directional strand. In some applied models, at least one of the directional strand nucleotides is an unmodified RNA nucleotide. In some applied models, at least one of the directional strand nucleotides is a 2'-deoxynucleotide (DNA). In some applied models, the remaining nucleotides of the directional strand are 2'-O-methyl-modified nucleotides. In some applied models, the anti-directional strand includes five fluoro-2' modified nucleotides, for example, at positions 2, 5, 7, 14, and 16 of the 5' end of the anti-directional strand. In some applied models, the anti-directional strand includes five fluoro-2' modified nucleotides, for example, at positions 2, 5, 8, 14, and 16 of the 5' end of the anti-directional strand. In some applied models, the anti-directional strand includes five fluoro-2' modified nucleotides, for example, at positions 2, 3, 7, 14, and 16 of the 5' end of the anti-directional strand. In some applied models, the anti-directional strand includes three fluoro-2' modified nucleotides, for example, at positions 2, 14, and 16 of the 5' end of the anti-directional strand. In some applied models, the other nucleotides of the anti-directional strand are 2'-O-methyl modified nucleotides. In some applied models, the 5' end of the strand against the direction contains a phosphate isotope, for example 5'-phenylphosphonate (5'-VP). In some applied models, the trend braid or anti-trend braid includes a non-stepping portion or an inverse non-stepping portion, for example, one of the portions shown in Table 6. Table 6. Non-stepping or inverse non-stepping parts ( iAb) Structure 1 (No step) 2 (iAb) "5'" and "3'" refer to the 5' to 3' direction of the sequences. In some applied models, the trending and counter-trend strands contain one or more bonds between modified nucleotides. In some applied models, the bond between modified nucleotides is a phosphorothiolate bond. In some applied models, the trending strand contains four or five phosphorothiolate bonds. In some applied models, the counter-trend strand contains four or five phosphorothiolate bonds. In some applied models, both the trending and counter-trend strands contain four phosphorothiolate bonds. And in some applied models, the trending strand contains four phosphorothiolate bonds and the counter-trend strand contains five phosphorothiolate bonds. Table 7 provides examples of modified directional and anti-directional strand sequences of dsRNA targeting human SARM1 mRNA. In some applied models, dsRNA includes a directional strand consisting of a sequence with one, two, or three differences from the directional strand sequence in Table 7. In some applied models, dsRNA includes an anti-directional strand consisting of a sequence with one, two, or three differences from the anti-directional strand sequence in Table 7. Table 7: Modified sequences from dsRNA Which targets SARM1 mRNA human number dsRNA braid Sequence from the end 5’ to 3’ Serial identification number 5 S mG*mU*mUmGmCmUmCmGfAfCfUmCmUmAmAmCmCmGmC*mU*mA 43 AS mU*fA*mGmCfGmGmUfUmAmGmAmGmUfCmGfAmGmCmAmAmC*mG*mG 44 6 S mU*mU*mCmGmCmCmAmAfCfUfAmUmUmCmUmAmCmGmU*mG*mA 45 AS mU*fC*mAmCfGmUmAfGmAmAmUmAmGfUmUfGmGmCmGmAmA*mG*mG 46 7 S mA*mC*mCmUmUmCmGmCfCfAfAmCmUmAmUmUmCmUmA*mC*mA 47 AS mU*fG*mUmAfGmAmAfUmAmGmUmUmGfGmCfGmAmAmGmGmU*mC*mU 48 8 S mC*mC*mGmCmAmAmGmAfGfGfUmUmCmUmUmUmAmGmG*mG*mA 49 AS mU*fC*mCmCfUmAmAfAmGmAmAmCmCfUmCfUmUmGmCmGmG*mG*mU 50 9 S mU*mU*mCmGmCmCmAmAfCfUfAmUmUmCmUmAmCmGmU*mG*mA 45 AS mU*fC*mAmCfGmUfAmGmAmAmUmAmGfUmUfGmGmCmGmAmA*mG*mG 51 10 S mU*mU*mCmGmCmCmAmAfCfUfAmUmUmCmUmAmCmGmU*mG*mA 45 AS mU*fC*fAmCmGmUfAmGmAmAmUmAmGfUmUfGmGmCmGmAmA*mG*mG 52 11 S mU*mU*mCmGmCmCmAmAfCfUfAmUmUmCmUmAmCmGmU*mG*mA 45 AS mU*fC*mAmCmGmUmAmGmAmAmUmAmGfUmUmGmGmCmGmAmA*mG*mG 53 12 S mG*mU*mUmGmCmUmCmGfAfCfUmCmUmAmAmCmCmGmC*mU*mA 43 AS mU*fA*mGmCfGmGfUmUmAmGmAmGmUfCmGfAmGmCmAmAmC*mG*mG 54 13 S mG*mU*mUmGmCmUmCmGfAfCfUmCmUmAmAmCmCmGmC*mU*mA 43 AS mU*fA*fGmCmGmGfUmUmAmGmAmGmUfCmGfAmGmCmAmAmC*mG*mG 55 14 S mG*mU*mUmGmCmUmCmGfAfCfUmCmUmAmAmCmCmGmC*mU*mA 43 AS mU*fA*mGmCmGmGmUmUmAmGmAmGmUfCmGmAmGmCmAmAmC*mG*mG 56 15 S mC*mC*mGmCmAmAmGmAfGfGfUmUmCmUmUmUmAmGmG*mG*mA 49 AS mU*fC*mCmCfUmAfAmAmGmAmAmCmCfUmCfUmUmGmCmGmG*mG*mU 57 16 S mC*mC*mGmCmAmAmGmAfGfGfUmUmCmUmUmUmAmGmG*mG*mA 49 AS mU*fC*fCmCmUmAfAmAmGmAmAmCmCfUmCfUmUmGmCmGmG*mG*mU 58 17 S mC*mC*mGmCmAmAmGmAfGfGfUmUmCmUmUmUmAmGmG*mG*mA 49 AS mU*fC*mCmUmAmAmAmGmAmAmCmCfUmCmUmUmGmCmGmG*mG*mU 59 18 S mG*mU*mUmGmCmUfCmGfAfCfUmCmUmAmAmCmCmGmC*mU*mA 43 AS mU*fA*mGmCmGfGmUmUmAmGmAmGmUfCmGfAmGmCmAmAmC*mG*mG 60 19 S mU*mU*mCmGmCmCfAmAfCfUfAmUmUmCmUmAmCmGmU*mG*mA 45 AS mU*fC*mAmCmGfUmAmGmAmAmUmAmGfUmUfGmGmCmGmAmA*mG*mG 61 20 S mA*mC*mCmUmUmCfGmCfCfAfAmCmUmAmUmUmCmUmA*mC*mA 47 AS mU*fG*mUmAmGfAmAmUmAmGmUmUmGfGmCfGmAmAmGmGmU*mC*mU 62 21 S mC*mC*mGmCmAmAfGmAfGfGfUmUmCmUmUmUmAmGmG*mG*mA 49 AS mU*fC*mCmCmUfAmAmAmGmAmAmCmCfUmCfUmUmGmCmGmG*mG*mU 63 Abbreviations – “m” stands for 2’-OMe; “f” stands for 2’-fluoro; “*” stands for phosphorothiolate binding; “S” stands for directional braid; “AS” stands for anti-directional braid; and unless otherwise noted, the 5’ position of AS can include 5’-phosphate or 5’-phenylphosphonate (VP). In some applied models, the directional and anti-directional strands in dsRNA involve a pair of DNA sequences selected from a set that includes the following: (a) The trending argument is identified by the serial identification number: 43, while the anti-trend argument is identified by the serial identification number: 44, or 54, or 55, or 56, or 60; (b) The trending braid is identified by the serial identification number: 45, while the anti-trend braid is identified by the serial identification number: 46, or 51, or 52, or 53, or 61; (c) The directional braid consists of the serial identification number: 47, while the anti-directional braid consists of the serial identification number: 48, or 62; and (d) The trending argument is identified by the serial identification number: 49, while the anti-trend argument is identified by the serial identification number: 50, or 57, or 58, or 59, or 63. In some applied models, the directional and anti-directional strands in dsRNA involve a pair of DNA sequences selected from a set that includes the following: (a) The directional braid is identified by the serial identification number: 43, while the anti-directional braid is identified by the serial identification number: 44, or 54, or 55, or 56, or 60; (b) The directional braid is identified by the serial identification number: 45, while the anti-directional braid is identified by the serial identification number: 46, or 51, or 52, or 53, or 61; (c) The directional braid consists of the serial identification number: 47, while the anti-directional braid consists of the serial identification number: 48, or 62; and (d) The directional braid consists of the serial identification number: 49, while the anti-directional braid consists of the serial identification number: 50, or 57, or 58, or 59, or 63. The oriented and anti-oriented strands of dsRNA can be synthesized using any of the well-known DNA polymerase methods in this field, for example, solid-phase synthesis using phosphomidesitization chemistry (e.g., Current Protocols in Nucleic Acid Chemistry, Beaucage, SL et al. (Edrs.), John Wiley & Sons, Inc., New York, NY, USA), hydrogen phosphonate chemistry, triphosphate ester chemistry, or enzymatic synthesis. Commercially available automated synthesizers, such as MerMade™ 12 from LGC Biosearch Technologies, or other syntheses from BioAutomation or Applied Biosystems, can be used. Phosphorothiazine bonds can be introduced using a sulfur reagent such as phenylacetyl disulfide or DDTT (((dimethylaminomethylidine)amino)-3H-l,2,4-dithiazolene-3-thione). It is well known that similar techniques, modified intermediate products, and commercially available controlled porous glass (CPG) products are used to synthesize modified oligonucleotides or conjugated oligonucleotides. Purification methods can be used to remove unwanted impurities from the final oligonucleotide product. Commonly used purification techniques for single-stranded oligonucleotides include reverse-phase ion-pair high-performance liquid chromatography (RP-IP-HPLC), capillary gel electrophoresis (CGE), anion-exchange HPLC (AX-HPLC), and size exclusion chromatography (SEC). After purification, the oligonucleotides can be analyzed by mass spectrometry and quantified by spectroscopy at 260 nm. The directional and antidirectional strands can then be annealed to form dsRNA. The RNAi factor described in this document can be prepared using a variety of procedures familiar to those with ordinary experience in the field, some of which are illustrated in the preparations and examples below, for instance, in Examples 1-3. Those with ordinary experience in the field understand that the specific synthetic steps for each of the described methods can be combined in different ways, or with steps from different schemes, to prepare the RNAi factor. The product of each step can be recovered using conventional methods well-known in the field, including extraction, evaporation, precipitation, chromatography, filtration, sintering, and crystallization. The reagents and starting materials are readily available to those with ordinary experience in the field. In some applied models, the TfR receptor-binding protein can be first treated with native or engineered cysteines, as described in this document, using a reducing agent, for example, dithiothreitol (DTT), and then re-oxidized using an oxidizing agent, for example, dihydroascorbic acid (DHAA). The resulting oxidized TfR receptor-binding protein is then incubated with dsRNA containing linker functional groups, for example, linker dsRNA, to produce the RNAi conjugate factor. Pharmaceutical formulation On the other hand, pharmaceutical formulations containing the SARM1 RNAi agents described here and a pharmacologically acceptable carrier are provided. These formulations may also include an excipient, a diluent, or one or more pharmacologically acceptable carriers. The formulations can be prepared using methods well-established in the field (e.g., Remington: The Science and Practice of Pharmacy, 23rd edition (2020), A. Loyd et al., Academic Press). Treatment method and therapeutic use On the other hand, there are methods available here to reduce axonal degeneration in a patient who needs it. This method involves giving the patient an effective amount of SARM1 RNAi factor or a pharmaceutical formulation described here. On the other hand, methods are available here to treat a SARM1-mediated neurological disease in a patient who needs it. This method involves giving the patient an effective amount of SARM1 RNAi factor or a pharmaceutical formulation described here. Examples of SARM1-mediated neurological 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), senile dementia, Pick's disease, Gaucher disease, Hurler syndrome, progressive multifocal leukoencephalopathy, Alexander disease, congenital hypomyelination, encephalomyelitis, acute disseminated encephalomyelitis, central pons demyelination, osmotic hyponatremia, Tay-Sachs disease, motor neuron disease, ataxia, spinal muscular atrophy (SMA), Niemann-Pick disease, acute hemorrhagic leukoencephalopathy, trigeminal neuralgia, Bell's palsy, cerebral ischemia, multiple system atrophy, Bleisius-Mertzbacher disease, periventricular leukomalacia, and ataxia. Genetic, noise-induced hearing loss, orCongenital hearing loss, age-related hearing loss, Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, Lewy body dementia, frontotemporal dementia, tau nephropathy, nucleopathy, amyloidosis, diabetic neuropathy, leukocyte atrophy (Krapp's disease), Bassen-Komzweig syndrome, transverse myelitis, motor neuron disease, spinocerebellar ataxia, preeclampsia, inherited spastic hemiplegia, spastic paraplegia, familial spastic paraplegia, French settlement disease, Strombel-Lorain disease, non-alcoholic steatohepatitis (NASH), adrenal spinal neuropathy, progressive supranuclear palsy (PSP), Friedreich's ataxia, spinal cord injury, acute optic neuropathy (AON), or a hereditary or unknown retinal condition The cause, or Leber congenital atrophy (LCA), or Leber hereditary optic neuropathy (LHON), or primary open-angle glaucoma (POAG), or acute angle-closure glaucoma (AACG), or dominant optic nerve atrophy, or retinal ganglion degeneration,Or retinitis pigmentosa, external retinal neuropathy, optic neuritis, multiple sclerosis-associated optic neuropathy, Kerr's optic neuropathy, ischemic optic neuropathy, chemotherapy-induced peripheral neuropathy, optic neuritis, Charcot-Marie-Tooth disease, vitamin B12 deficiency, folic acid (vitamin B9) deficiency, isolated vitamin E deficiency syndrome, non-arteritic anterior ischemic optic neuropathy, ethambutol exposure, cyanide exposure, traumatic brain injury (TBI), spinal cord injury, traumatic axonal injury, chronic traumatic encephalopathy (CTE). In some applied models, SARM1-mediated neuropathy is amyotrophic lateral sclerosis, multiple sclerosis, chemotherapy-induced peripheral neuropathy (CIPN), diabetic peripheral neuropathy (DPN), tau neuropathy, or Charcot-Marie-Tooth disease. In some applied models, SARM1-mediated neuropathy is a neurodegenerative disease of the central nervous system. In some applied models, SARM1-mediated neurological disease is amyotrophic lateral sclerosis (ALS, or Lou Gehrig's disease), Alzheimer's disease, Parkinson's disease, multiple sclerosis (MS), Huntington's disease (HD), Pick's disease, progressive multifocal leukoencephalopathy, Alexander disease, congenital hypomyelination, encephalomyelitis, acute disseminated encephalomyelitis, central pons demyelination, Tay-Sachs disease, spinal muscular atrophy (SMA), cerebral ischemia, Lewy body dementia, frontotemporal dementia, tau disease, syncylinopathy, amyloidosis, diabetic neuropathy, spherocytic leukodystrophy (Krappa disease), Strombel-Lorain disease, adrenal spinal neuropathy, progressive supranuclear palsy (PSP), or Leber's hereditary optic neuropathy. (LHON), or optic neuritis, or optic neuritis associated with multiple sclerosis, or Kerr's optic neuropathy, orIschemic optic neuropathy, chemotherapy-induced peripheral neuropathy, myelitis optica, traumatic brain injury (TBI), spinal cord injury, traumatic axonal injury, or chronic traumatic encephalopathy (CTE). In some applied models, the SARM1-mediated neurological disease is amyotrophic lateral sclerosis. SARM1 RNAi factor or a pharmaceutical formulation containing SARM1 RNAi factor can be given to the patient intravenously or subcutaneously. The dosage regimen for SARM1 RNAi can be modified to provide the desired optimal response (e.g., therapeutic response). For example, a single bolus can be administered, several doses can be given over time, or the dose can be decreased or increased proportionally as indicated by the therapeutic requirements. Dosage values ​​may vary depending on the type and severity of the condition being treated. It is also understood that for any given study participant, the prescribed dosing regimens should be adjusted over time according to individual needs and the professional judgment of the person taking or administering the formulations.On the other hand, SARM1 RNAi agents or pharmaceutical formulations containing SARM1 RNAi are provided for use in reducing SARM1 expression. On the other hand, SARM1 RNAi agents or pharmaceutical formulations containing SARM1 RNAi are provided for use in reducing axonal degeneration. SARM1 RNAi agents or pharmaceutical formulations containing SARM1 RNAi are also provided for use in treating SARM1-mediated neurological disease. The uses of SARM1 RNAi agents in the manufacture of a drug to reduce axonal degeneration are also provided. The uses of SARM1 RNAi agents in the manufacture of a drug to treat SARM1-mediated neurological disease are also provided. Definitions As used in this document, the use of indefinite and definite articles and the like in the context of the present disclosure (especially in light of the following safeguards) includes both plural and singular, unless otherwise indicated or clearly contradicted by the context. As used here, the term "alkyl" refers to a linear, saturated, or branched-chain monovalent hydrocarbon radical containing the specified number of carbon atoms. For example, "C1-C alkyl" 20 "To the presence of a root containing 1-20 carbon atoms in a linear or branched arrangement. The term "antibody," as used in this document, refers to a molecule that binds to an antigen. Applied antibody models include monoclonal antibodies, polyclonal antibodies, human antibodies, human-compatible antibodies, chimeric antibodies, heterodimeric antibodies, bispecific or multispecific antibodies, and conjugate antibodies. Antibodies can be of any class (e.g., IgG, IgE, IgM, IgD, IgA) and any subclass (e.g., IgG1, IgG2, IgG3, IgG4). The immunoglobulin G (IgG) antibody consists of four polypeptide chains: two heavy (HC) and two light (LC) chains cross-linked via disulfide bonds between the chains. The amino-terminal region of each of the four polypeptide chains contains a variable region of approximately 100–125 or more amino acids, primarily responsible for antigen recognition. The carboxyl-terminal region of each of the four polypeptide chains contains a constant region, primarily responsible for effector function. Each heavy chain consists of a variable heavy chain (VH) region and a constant heavy chain region. Each light chain consists of a variable light chain (VL) region and a constant light chain region. The IgG isotype can also be divided into subtypes (e.g., IgG1, IgG2, IgG3, and IgG4). The VH and LCVR regions can also be subdivided into hypermutable regions, called co-determinative regions (CDRs), interspersed with more conservative regions, called frame regions (FRs). Co-determinative regions are located on the protein surface and are crucial for antigen-binding specificity in the antibody. Each VH and VL consists of three CDR regions and four FR regions, arranged from amino-to-carboxylic end in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. In this document, the three heavy-chain CDRs are referred to as "HCDR1, HCDR2, and HCDR3," and the three light-chain CDRs are referred to as "LCDR1, LCDR2, and LCDR3." The CDRs contain most of the structural units that form specific interactions with the antigen. The structural amino acid units can be allocated to specific regions for completeness according to known schemes, including those described in Kabat (Kabat et al., "Sequences of Proteins of Immunological Interest," National Institutes of Health, Bethesda, Md. (1991)), Chothia (Chothia et al., "Canonical structures for the hypervariable regions of immunoglobulins", Journal of Molecular Biology, 196, 901-917 (1987); Al-Lazikani et al., "Standard conformations for the canonical structures of immunoglobulins", Journal of Molecular Biology, 273, 927-948 (1997)), North (North et al., "A New Clustering of Antibody CDR Loop Conformations", Journal of Molecular Biology, 406, 228-256 (2011)), أو IMGT (the international ImMunoGeneTics database available on at www.imgt.org; راجع Lefranc et al., Nucleic Acids Res. 1999; 27:209-212). The current detection application models also include antibody fragments or antigen-binding fragments, which, as used in this document, include at least one antibody fragment that retains the ability to react specifically with the antigen or antigen epitope such as Fab fragments, Fab', F(ab')2, Fv, scFv antibody fragments, scFab fragments, disulfide-bound Fvs fragments (sdFv) and Fd fragment. The term "antigen-binding domain," as used in this document, refers to a part of an antibody or an antibody fragment that binds to an antigen or an epitope of an antigen. For example, the term "TfR receptor-binding domain" refers to a part of an antibody or an antibody fragment that binds to a TfR receptor or an epitope of the TfR receptor. The term "heterogeneous dimeric antibody," as used in this document, refers to an antibody consisting of two different antigen-binding domains. As used here, the term "anti-directional strand" refers to a single-stranded oligonucleotide complementary to the target sequence region. As used here, the term "directional strand" refers to a single-stranded oligonucleotide complementary to the anti-directional strand region. The terms “bond” and “associate” as used in this document, unless otherwise stated, refer to the ability of a protein or molecule to form a chemical bond or attractive interaction with another protein or molecule, resulting in the affinity of the two proteins or molecules as determined by common methods known in this field. As used in this document, the term "complementary" refers to the structural relationship between two nucleotides ( For example In two opposing DNA sequences or in opposing regions of a single DNA strand, example (A bent container) allows two nucleotides to form base pairs with each other. For example, a purine nucleotide from one DNA molecule complementary to a pyrimidine nucleotide from an opposing DNA molecule can form a pair by forming hydrogen bonds with each other. Complementary nucleotides can form a pair via the Watson-Crick method or by any other method that allows for stable pairings. Similarly, two DNA molecules may contain regions of multiple nucleotides that complement each other to form complementation regions, as illustrated here. As used here, the word "pair," when referring to nucleic acids or oligonucleotides, means a structure formed by the complementary base pairing of two parallel (i.e., opposite-oriented) sequences of nucleotides, whether composed of two separate DNA strands or a single folded strand ( For example (via a bent vessel). The term "effective dose" refers to the amount required (for specific durations and routes of administration) to achieve the desired therapeutic outcome. The effective dose of a protein or conjugate can vary depending on factors such as the patient's condition, age, sex, weight, and the protein or conjugate's ability to elicit the desired response. The effective dose is also defined as the amount at which any toxic or harmful effects of the protein or conjugate are outweighed by its therapeutically beneficial effects. The term "Fc region," as used in this document, refers to a polypeptide containing the CH2 and CH3 domains of a stable region of immunoglobulin, for example, IgG1, IgG2, IgG3, or IgG4. Optionally, the Fc region may include part of the hinge region or the entire hinge region of immunoglobulin, for example, IgG1, IgG2, IgG3, or IgG4. In some applied models, the Fc region is the Fc region of human IgG, for example, the Fc region of human IgG1, the Fc region of human IgG2, the Fc region of human IgG3, or the Fc region of human IgG4. In some applied models, the Fc region is the Fc region of a modified IgG with reduced or no response functions compared to the corresponding Fc region of wild-type IgG. The numbering of residues in the Fc region is based on the European Union index as described in the Kabat et al. numbering. Sequences of Proteins of Immunological Interest (Protein sequences of immunological importance) (5th ed., Bethesda, MD: U.S. Department of Health and Human Services, Public Health Services, National Institutes of Health, 1991). The boundaries of the Fc region of the immunoglobulin heavy chain may vary, and the Fc region of the human IgG heavy chain is usually defined as the extension from the N end of the CH2 domain (e.g., the amino acid residue at position 231 according to EU index numbering) to the C end of the CH3 domain (or the C end of the immunoglobulin). The term “decline” or “decline in expression” refers to a decrease in the gene expression of mRNA or protein following treatment with a reagent. As used in this document, the term "modified internucleotide bond" refers to an internucleotide bond that contains one or more chemical modifications when compared to a reference internucleotide bond containing a phosphate diester bond. The modified internucleotide bond may be an unnatural bond. In some applied models, the modified internucleotide bond is a phosphorothiolate bond. As used in this document, the term "modified nucleotide" refers to a nucleotide that contains one or more chemical modifications when compared to the corresponding reference nucleotide chosen from: adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide, uracil ribonucleotide, adenine deoxyribonucleotide, guanine deoxyribonucleotide, cytosine deoxyribonucleotide, and thymidine deoxyribonucleotide. A modified nucleotide may, for example, contain one or more chemical modifications in the sugar group and / or the nucleobase and / or phosphate group. Additionally, or alternatively, a modified nucleotide may contain one or more chemical elements associated with the corresponding reference nucleotides. In some applied models, the modified nucleotide is a 2'-fluoro-modified nucleotide, a 2'-methyl-O-modified nucleotide, a 2'-deoxynucleotide (DNA), or a 2'-Alkyl-O-modified nucleotide (e.g., alkyl 2'-OC). 16 In some applied models, the modified nucleotide contains a phosphate isotope, for example, 5'-phenylphosphonate. In other applied models, the modified nucleotide contains a non-stepping or inverse non-stepping motif, for example, the motif shown in Table 6. As used here, "nucleotides" refers to an organic compound containing a nucleoside (a nucleic acid base, for example, adenine, cytosine, guanine, thymine, or uracil, and a pentose sugar, for example, ribose or 2'-deoxyribose) linked to a phosphate group. A "nucleotide" can be a single unit of nucleic acid polymers such as deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). As used in this document, "empty arm" means the arm of antibodies that does not bind to any known human target. As used here, "oligonucleotide" refers to a polymer of linked nucleotides, each of which may or may not be modified. An oligonucleotide is typically less than about 100 nucleotides in length. As used here, the term "buffer" refers to the nucleotides or unpaired nucleotides that protrude from the doublet structure of a double-stranded oligonucleotide. A buffer may consist of one or more unpaired nucleotides extending from a doublet region at the 5' or 3' end of a double-stranded oligonucleotide. A buffer can be a 3' or 5' buffer in the counter-strand or directional strand of a double-stranded oligonucleotide. The term "patient," as used here, refers to a human patient. As used here, the term "phosphate isotope" refers to a chemical component that mimics the electrostatic and / or stereostatic properties of a phosphate group. In some applied models, the phosphate isotope is placed at the 5' end of the oligonucleotide instead of the 5'-phosphate, which is often susceptible to enzymatic removal. The 5' phosphate isotope may contain a phosphatase-resistant bond. Examples of 5' phosphate isotopes include methylene phosphonate (5'-MP) and 5'-(E)-phenylphosphonate (5'-VP). In some applied models, the phosphate isotope is 5'-VP. As used here, "SARM1" (sterile alpha-and-TIR motivator containing 1), also known as SARM; HsTIR; SAMD2; hSARM1; MyD88-5, refers to the human SARM1 mRNA transcript or human SARM1 protein. The nucleotide sequence of human SARM1 mRNA can be found at NM_015077.4: 1 ATCTCCCAGC TCAGCCGAGC CCGTGCCCAG GCCACGCTTT GTTCCAGCCG CCGCCTCCTC 61 TACCCTACGG CGTCCGGAGC CATCCCTCGC CTGCTCGCTC TCTCCTTTCG CCCACTCCCT 121 GCATCTGGGC CTGCATCACC TTTGCCAACC GCTCCCCCGA TCCTGCCGAC ACTCCTCCCC 181 CAAACTTCTG ACCGGCACCC TTGCCTGGTA CCCTTCTCTC CATTCCTCCC CCTCCATCTT 241 CTTTCCCCGA CCCCTCTCGG GTCCCTCTTT TCCCAAAACC CGGGTCTCTC CGCGTGGCCC 301 CGCCTCCAGG CCGGGGATGT CCCCCGCGGC CCCGCGCCCA TGGTCCTGAC GCTGCTTCTC 361 TCCGCCTACA AGCTGTGTCG CTTCTTCGCC ATGTCGGGCC CACGGCCGGG CGCCGAGCGG 421 CTGGCGGTGC CTGGGCCAGA TGGGGGCGGT GGCACGGGCC CATGGTGGGC TGCGGGTGGC 481 CGCGGGCCCC GCGAAGTGTC GCCGGGGGCA GGCACCGAGG TGCAGGACGC CCTGGAGCGC 541 GCGCTGCCGG AGCTGCAGCA GGCCTTGTCC GCGCTGAAGC AGGCGGGCGG CGCGCGGGCC 601 GTGGGCGCCG GCCTGGCCGA GGTCTTCCAA CTGGTGGAGG AGGCCTGGCT GCTGCCGGCC 661 GTGGGCCGCG AGGTAGCCCA GGGTCTGTGC GACGCCATCC GCCTCGATGG CGGCCTCGAC 721 CTGCTGTTGC GGCTGCTGCA GGCGCCGGAG TTGGAGACGC GTGTGCAGGC CGCGCGCCTG 781 CTGGAGCAGA TCCTGGTGGC TGAGAACCGA GACCGCGTGG CGCGCATTGG GCTGGGCGTG 841 ATCCTGAACC TGGCGAAGGA ACGCGAACCC GTAGAGCTGG CGCGGAGCGT GGCAGGCATC 901 TTGGAGCACA TGTTCAAGCA TTCGGAGGAG ACATGCCAGA GGCTGGTGGC GGCCGGCGGC 961 CTGGACGCGG TGCTGTATTG GTGCCGCCGC ACGGACCCCG CGCTGCTGCG CCACTGCGCG 1021 CTGGCGCTGG GCAACTGCGC GCTGCACGGG GGCCAGGCGG TGCAGCGACG CATGGTAGAG 1081 AAGGCGCGCAG CCGAGTGGCT CTTCCCGCTC GCCTTCTCCA GCTGCTTCGG 1141 CTGCACGCCT GCCTCGCAGT AGCGGTGTTG GCGACTAACA AGGAGTGGA GCGCGAGGTG 1201 GAGCGCTCGG GCACGCTGGC GCTCGTGGAG CCGCTTGTGG CCTCGCTGGA CCCTGGCCGC 1261 TTCGCCCGCT GTCTGGTGGA CGCCAGCGAC ACAAGCCAGG GCCGCGGGCC CGACGACCTG 1321 CAGCGCCTCG TGCCGTTGCT CGACTCTAAC CGCTTGGAGG CGCAGTGCAT CGGGGCTTTC 1381 TACCTCTGCG CCGAGGCTGC CATCAAGAGC CTGCAAGGCA AGACCAAGGT GTTCAGCGAC 1441 ATCGGCGCCA TCCAGAGCCT GAAACGCCTG GTTTCCTACT CTACCAATGG CACTAAGTCG 1501 GCGCTGGCCA AGCGCGCGCT GCGCCTGCTG GGCGAGGAGG TGCCACGGCC CATCCTGCCC 1561 TCCGTGCCCA GCTGGAAGGA GGCCGAGGTT CAGACGTGGC TGCAGCAGAT CGGTTTCTCC 1621 AAGTACTGCG AGAGCTTCCG GGAGCAGCCAG GTGGATGGCG ACCTGCTTCT GCGGCTCACG 1681 GAGGAAC TCCAGACCGA CCTGGGCATG AAATCGGGCA TCACCCGCAA GAGGTTCTTT 1741 AGGGAGCTCA CGGAGCTCAA GACCTTCGCC AACTATTCTA CGTGCGACCG CAGCAACCTG 1801 GCGGACTGGC TGGGCAGCCT GGACCCGCGC TTCCGCCAGT ACACCTACGG CCTGGTCAGC 1861 TGCGGCCTGG ACCGCTCCCT GCTGCACCGC GTGTCTGAGC AGCAGCTGCT GGAAGACTGC 1921 GGCATCCACC TGGGCGTGCA CCGCGCCCGC ATCCTCACGG CGGCCAGAGA AATGCTACAC 1981 TCCCCGCTGC CCTGTACTGG TGGCAAACCC AGTGGGGACA CTCCAGATGT CTTCATCAGC 2041 TACCGCCGGA ACTCAGGTTC CCAGCTGGCC AGTCTCCTGA AGGTGCACCT GCAGCTGCAT 2101 GGCTTCAGTG TCTTCATTGA TGTGGAGAAG CTGGAAGCAG GCAAGTTCGA GGACAAACTC 2161 ATCCAGAGTG TCATGGGTGC CCGCAACTTT GTGTTGGTGC TATCACCTGG AGCACTGGAC 2221 AAGTGCATGC AAGACCATGA CTGCAAGGAT TGGGTGCATA AGGATTGT GACTGCTTTA 2281 AGCTGCGGCA AGAACATTGT GCCCATCATT GATGGCTTCG AGTGGCCTGA GCCCCAGGTC 2341 CTGCCTGAGG ACATGCAGGC TGTGCTTACT TTCAACGGTA TCAAGTGGTC CCACGAATAC 2401 CAGGAGGCCA CCATTGAGA GATCATCCGC TTCCTGCAGG GCCGCTCCTC CCGGGACTCA 2461 TCTGCAGGCT CTGACACCAG TTTGGAGGGT GCTGCACCCA TGGGTCCAAC CTAACCAGTC 2521 CCCAGTTCCC CAGCCCTGCT GTGACTTCCA TTTCCATCGT CCTTTCTGAA GGAACAGCTC 2581 CTGAAACCAG TCTCCCTGGG CTGAGACAAC CTGGGCTCTT CTTAGGAAT GGCTCTCCCT 2641 CCCCCTGTCC CCCACCCTCA TGGCCCACCT CCAACCCACT TTCCTCAGTA TCTGGAGAGG 2701 GAAGGGAAGT CAGGCTTGGG CACGGGAGGT TAGAACTCCC CCAGGCCCTG CCATTGGGTT 2761 GTCTGTCTCC GTCATGGGGA GGGTCCCTGC TCAGTTCTGG AGACACTGGA GTTGGGGTGG 2821 GGGTGGTTCT GCATTCCCTT CTCCTGCTGA TAGCAGTCAG CTTGAGGAGG ATGACGGAAG 2881 GCAGCCTCAG ACAGGAATTA AGGCAATGCC CAGGCGGGCC TGGGCACTGT ATTCTGAGCA 2941 AGGGCCTGGG CCCAGGAGCC AGCCAGGGAT GAGTGCCATC ATGGCTCTCC ACTCAGACTG 3001 TGCCTGGCCC CTGCACTTAC AACTTCCTGC CGCTCTGTGG CCTTGCCCTG TAATCACTCA 3061 GTGCCCTTAG CTAGCCTGAC TAAGTCCCAG ATCCCCTACA GCTTCCTTCG GTGTGGTATC 3121 TTTTGCCACA TCCAGGGCGA GGGTTGAGGC AAACCAGCCC TCCCTCTGAC TTCCTTGTCA 3181 CTGCAGCCAG CTTTGCTGCA CTTGCTGGTG CACAGGAGCC TCCTGTTTGG GCCTGGGTCT 3241 GGGCATGGGG AGGCCGTGCC TCAAAGCCCA CCCTACCCCA TGCCTTGGTG CTGTGCCTCA 3301 GGCTCCTTCC TGGTCTGGCC CAGCTGGCTT CCCCAGCCCC TCAGCCATCC AGGGCTACCC 3361 ACTGCTTACT CAGGGACCAG GCAGCCCCCA TGGCAGTAAA AGCAGCCTAG ACAGAACCTG 3421 CAGCTCTGTG GAAAGAGGCA AAGTCCTGAA AAGGCAAAGG GTTGTCACTT AGGGCAGCTT 3481 CTCCAACTTT AACATGCATC CAAGTCACCT GGGAATGTTG TTAAAATCAG GAGATCTGGG 3541 GTGGGGCCTA GGACTCTGCA TTTCTTACAG ATTCCCAGGT GAGCTGATGC TGGTGGTTAA 3601 GGGTAGCAAA TCTCTAAAGC ACGAAGCCCT CACAAATCTT TGCCATTTCC CAAACACTCC 3661 GCTCCATGGT CTCCAGTCAT CAGAGCAACT CTACCTGGTA TTATCATCCC CATTTTACAG 3721 ATAATGACAC TGAGGCTCAG AAAGGTTGAG GATAAGCCCA CTTTCCTGTC ATTAGTGGCA 3781 GCCCCAGATC CAGACCTAGG CCTCCTGGCA CCCAGTCCAC TGGCAGTGGA ATTGCTTTCC 3841 TGAGAATCAT TCTGAGGCTG GGCTATTGCT TCTCCCTTGC TTCAAAGAAT CTAGCAGCGG 3901 GGGATAGGAT TTTGCAACAA AAAGCTGACC CAGAGGCCAT ACAGAGCAGG AATATCCCAT 3961 TGCCCCCTCC TCCACTGGGT TCAGAGGGTA AGAAAGCACC CTCCAATAAA CCCAGGCTCC 4021 AGGCCGTGGG GGCTGCTGAA GGCTCTTTCC CCGCAAGGGC CAGGTGTTGA CACCTTAAAG 4081 CTGGCTGCGC CCCCAGCCCC ACTCTTGGCT GTGCTGGCCA GGTGACTCCT AGTTCTTGGC 4141 CACATCATCA GAAAGTCAAA GGTCTCACTC CAGGTTTGGG GCTCCTTCCT TCCACTCCCC 4201 TCCCTGCCAG AGTCTGTCTT GGCCAGTGCC AGCCTCGATG CTTTGGTTTT GACCCCACCT 4261 GATCCTCCTT TCCTCATGCA GCACAAGTGC TCACCGGGGC CAGAGCCAGG GCATGGATAT 4321 GACAAGCAGG GCAGCCTGGA CACTGCCCTC ACAGGACAGC GCCAATAACA ATACAGTGTC 4381 TGAGTATCTC CAGGGGATGA TTTCTGGCTC TTTGTCTCCA ATCAGTCCCA CTCCCTCCTG 4441 AGGTCCCCAA GGGCAGTATT CAGAGAGGTT TCCTGCGTTT TATTTCTATT TGGTATACCC 4501 TCCACTGTTG TCCACTGCCC TGTGTGGCCT TCTGGTTGAC CTCTGCCCGA TCTTCTGTCT 4561 CTCTGAGGGA ATCAGAGTCC AGCATCCAGC CCCAGCTGGA ACAGCTGAAG TCACAAGCCT 4621 CCTCTAAGCC AAGGCCAGTG TGTTCAGAGG TGACTGCCAC CCATACTAGG ACAAACACAG 4681 CTCAGATCAC CAGGTCAAGC ACCTAGGCCT GGCTTCTCCT GAGACAGAGG ACTCAGAAGT 4741 GGCCTTTCCT CCAAAGCCTG CTCAGACACA GGTCTGTAGG GCCAGGGTGT TCTGCTTGGC 4801 TGGGCTGCAG CTGCTACCCC TCGGTTGGGG CTGAGTCAGC CAGATCCTCC CCCTACTTCT 4861 CCCCAAGGGC CAAGAACTGC TCAGGGACAT TAAAGGTCAA AAGTCCAGCC ACACTCATTC 4921 ATCCTTTCCC CAGGCCCATG AAGAGAGGCA TCTCATTGTA GAATGTATGA GGAAGTGGGGA 4981 AGTATCTCAG AGAATCAGCT AAGTTTCCTA ACTTGTCCAT CCAAATGTGA TCACCACGAT 5041 TCAACAATTT GGGGCATTGC TGATCTAGCC GTTCCTAGTG GGGCTTGCTC AAGGTTGCAC 5101 AGCGAGTCAG TAGAAGCCCT GGCTGGCCCC ACTTGGTACC AATCCACCAG GCAGCTCAGG 5161 GCTCCTGCCC AGCCCAGCAG CTTCTGTTGT CTAACGTATG GCAGGCAGAC TGGGAGCAGG 5221 AAAACAGAGG GCCCCAAAGC CCAAGGCACC AGAAGGTTTG TTTCAGTTTG CTGAAGCTGA 5281 TTTGTAATGA TTGGCACTCT TCAGCCAGGG GAGTGGGTAG GCCATAGCCA AGGATCGATT 5341 CCCCAACCAC AGCAAAGGCA ACACTCTTCC TCCAGAGATC ACCAAGCCCC TCTTACCTCC 5401 CTCCCTCCTT CCCAAGGCTG GCACTAACCA GGTACCACAT TCATTGTTAA GGAATGGCTG 5461 ATGACTGCTA CACGTGTTGG GAACCTGGTT GGGGCTGTGC AGTTTGGGCT GGAAGGAGAG 5521 ATGCCAGCCC TCGTGCTGCC TCTGGTCCCT GAAGTGTCAC CTCTCTCAGG ACCTCTCCTC 5581 TGGCCTGTGG GGTTATAAGT GATGGATAGC AGAAAGGGAG AACTGACTCC TGTCCCAAAT 5641 AGCTCCTCTG CCACCTGTCC TGCAGTGGGC CTGTGTGGGT TATGATTCTA GATCCTAGAC 5701 AGAGGCTGGG TCAGCTGTGG ATGGGGTGGT GCCTTGGTCT CTCTTGACTA CCTCGTCCAA 5761 AGAGAGCACT GCCCTTAGAC AAGAGTTGCT TGTCCTGCTG TGGGCTGGGC TTCCAGCTGC 5821 AGACCTCCAG TTGCTTGGTG TTCACTTTGC TCCTCTTGCC CTCTGTCTTC TGGTCCAGGC 5881 AGATCAGGGG CTCTGGGGAA ACTGCTGGAA CTCGAGGTGA GGATCAGCCT TTTCCAGCAT 5941 CCTGTGAGAG ACCAGAGAGA GAGTTTGGAT TTCATGTGGG GAACCCTCAA GGCCTGTCTG 6001 GAGAAGTGAC ACAGGATTTA CTGGGGTGGG CTGGTCCAGG TAGCTCTCCT GAACCTCCTC 6061 CTTCCCCAAG CTGAGAAGCT GAGAGCTGGA GGACAATATC CAGGGACATG GCTCTGGAAA 6121 ATAACTTTTT TTTTTTTAAG AGACAGGGTC TTGCTCTGTT GTCCAGGCTG GAGGGCAGTG 6181 ACATAATCAT AGCTCACTGT ACCCTTGAAC TCCTGGGCTC AAGTGATCCT CCTGCCTCAG 6241 CCTCCTTAGT AGCTGGGACT ACCAGTGCAT ACCACCATGC CTGGGTGATT TTTTAAATTT 6301 TTTATACAGA CAAGGTCTTG CTATGTTGCC CAGGCTGATC TTGAATTCCC GGGCTCAAGT 6361 GGTCCTCCTG CCTCAGCCTC CCACAGGATC GGGATTACAG GCAAGAGCCT CCACGCCCGG 6421 CCATGAAATA TAATTCTTAA TATCATACAG GAAAAAGTCA GCGGGTCAAG CTAGCCTGTG 6481 GCCCAGCCAC AACTAGCTGA CAAAGCTTCC TGGCCTTCCC TTTAACACAG TTCTGCTGCCC 6541 ATAGTTCCAT CTATAAAATG GGAATGGAGG GAAATAGGGG AACTGGGA GAGAACACAG 6601 CCTTGCCAAG CAGCAATGTT AGCCTGATCC TTCCTCCACC TAGCTCGCCA TCTCGCCCTT 6661 GGAAAATGGC TCCTGGAGGA TTAGGCAGCC ATCTGCAAGG AGAGGGGCAA CCTGGGACAA 6721 GACACCCAGA GGGTAAGGAT TCCAGGAATG AAGCTGCCAT TTCTGGTTGG GAGGAGAGA 6781 GGAAACTTTT AAGAGAAAGG GCTCCATTAT GAGCATGGGT TCAGGGCCCT GCATTACCCA 6841 ATCAGAACAG CCGGGATGAG CAGGAGGCCA GCTCCCAGGA GGAAGGGGAA CCCCTTCATA 6901 AAGTTCAGAG TGGCTGGGTA GAGTGAGTTG AAGATGCCGG AGGCCGTCAG CATGGCCAGG 6961 CTATTCACAC AGGCCACAGC AGAAAAAGAGA GCACCTGTGA AGAAATAAAT ACCATACTCT 7021 GGAGTCCGAA AGGGCCATAT TCCAACTCTG GCACCACCAC CTCACAGCTG TGTGACCGGG 7081 AGTAGTCACT TAACCTATGT CTCCCCTTCC TCACCAGTAA ATCCTGCTAC ATCATGTACT 7141 GTGACAAGGA TTCAGTAAGG TCATATGTGG ACAGTAGCTG GCACAGAGGG GCTACTAAAC 7201 AAATGGCTGC TATTAAATCC ACATTAAAAG TACATGTGAT CTHEADAC CCHEAD 7261 AAAGAAAAAAA AAAGTACATG TGATATTGTC TGATGAAAGC TTGATGGAAA TGGCTTTTTT 7321 CTGGTTTATC CTCTTTGGAA TCATTCCTG TTTGGGATTA ACTGCTGGTC TGATCAGTTC 7381 CAATATTCAT AGCGGTGTCA CCACTGAATA GCTTCTTATC CTTTGGGTTC CTGTTCCTCC 7441 TTCTGCTAAA GROUP ACCTATTTCC GROUP GTGA GCAACTTAA GTTCACATGG 7501 AAATCACCCA TCACTGGGCC TGGTCCCCTG GAAGTAGCTA GTTAGTAAGG GCTGTTCTTT 7561 TCTCCTGTTT CTCTTGACAT CTCTGGGCAC AGAGAAAGTG CTGGGAAAAA AAGTTTAGGT 7621 EXTENSION CAPACATGG ATTCTGGGGA CAPACITY CAPACITY GGGC TCTGTATGGC 7681 ACCAGAGTCT CTGTCATCAT CAGATCCTCA TTCCAGGACA GATGGAAAAA GATGATGTT 7741 TCCAGACTGG GGCATAAAGA CCCAGAGGCT GGAGAAGCTG TTCTTTATAG ATATACCCAGG 7801 AGAACCCACA GTTTACAAAA TGTGCAACAA CCCAACAGAA GTTGAGATTA AATTCTGTCA 7861 CATCTAGAGG GGTCTGTGAT GTCATCAAAA GCAAACCACC CACATCACAG ATGAAGAAAC 7921 AGGCCTGTGG CAGGGCTCGG ACTAAAACCC AGATCCTGAG ACCAGCTGCT TTTAAACACA 7981 GACGTAGGTT TGCATCCTAG CTCCACCATT TACTGAGTAA CCTTGGGTGA GCCAATGTAA 8041 CCCCCTGGGT CTCTGTTTCT TTATCTGTCA ACTGTGGAAA ATGAAACCCA TGTCACAAGG 8101 TTGTTCACTT CTGGGCTTGT ACACGCTGAC CCCAGAGAAA CAGGGAACTC TGGCATCACC 8161 ACACCCATCT TACAGACGGA AAAGCTGAG TCTGCAGAGA GTAAATCCTC TGCTCTGGTT 8221 ATCTAGAAAG AACATAATTG TGCTCTGCTG ACTGCAAATC CCAACTCTGC GGTTTGAAAA 8281 TCCAAGGTGG CATGATCCTC TGCCCATTGT GGGCAATTTC ACAGAAATGT GTTTGTTTTG 8341 GCCACTTACT TCTCCAGGGT GAGAGGGGGG AAGGCAAGCT GTTCCCCCAG CCATGGCTGC 8401 CCATCAGCCC GTTTCGGGCA GCACTGGACA TGAGGAACCA GACACAGGTG GGTTCTGACA 8461 CTCACCCTGC TCGTTCTCTC TCACCAGCTT GGAGAGTTTA GCCCGGATGA CAGGTGTGAT 8521 GACTAATGAC AGGAAAAGCA ACCCATATCC TGTGGAGAAA CAAACACTCA TCAGGAAAAT 8581 GGGGCTGGGG AGAGGGGCGT CCAAGGGAAA GGCAGCAGAG CTCCTATCCA TACCCCACGT 8641 GGGGCTTAGG TTAGACCCAG GAAGAACTTC CTTGATGGTG AGGGTGGGAA GACAGTAGTC 8701 AAGGAGGAAT GGAGACTGCC CTTGTCTGGG CTTGGCCACC TGCTAGCTCT CATGAATGAA 8761 TGCTAATTTCC CATTGATTGC TTTCTTGTCT GAACCTCTTG TGGTCACAGC AGGCATCACC 8821 CACCACTTG GCACTTAGTA GGGATATGGC AGGGCACAGA AAACAAGCAT GGGCTTTGGA 8881 GTCAGCCCTG AGTTCAAAAC CTGATGCCAT TACATATTAT CTGTGTGGCC TGGGGTACTT 8941 ACCCTCTCTG ATCCTGACTC CCTGTATGAG GAAGATAATA AGGCCTTCAT CACAGGATGG 9001 TTCTGAGGCA TAGGAGGCTG AATAATGGTG CCCAATGGCA TCAGATTCAT AGCCCTGGAA 9061 CCTGTAAATA CTACCTTATT TGGAAAATGA GTCTATGCAG GTGTGCAGTT AAGCCTCCTG 9121 CONFUSION GTTATCCTGG ATTAGGTTGG GCCCTAAATG CCGTCACACA TATCTTTATA 9181 AGAGGAAAGC AGACGGAGAT TTGGCACCGA AGATTGAG AAGGCACAAA GAGGAGGAGA 9241 GTCAATGTGA GCACAGAGGC AGAGACTGGT GATGGCCGCC CCAAGCCAAG GAATGCCAGC 9301 AGCCCCAGAA GCTGGAAAA ATGAAACA CGTTCTCTCC TGGAGGCTTG CAUGHT 9361 CTGCCTGCTG ACTGCTTCCA TTCAGCCCGG TGGTACTGAC TTTGGACTTC TGGCCTCCAG 9421 AACTGTGAGA GAATATGTTT CTGTTGTGTT AAGCCCCAA GTTTGTGGTA TGTCATTACA 9481 GCAATCTCAG GGAACCAATA CATGAGGTAA AAAGGTAACA TCTATGAAGA GCATGGCATA 9541 GGGACACAGC AAATGGGAGT TCCTTTTCCC TTTGCATTCA GTTACTTACA GGCTTCCTGT 9601 TTTCTTCATA ACCATTCTC TCCCTGTGCG ACTGCTGACT CCTCAGCAAA ACTGCAAACT 9661 CCTACAGGAC AGTGGATCCT CCAAAGAAGG TATACGATGA GGCATCCAGG GACCCTAGCA 9721 GTGTCAGGCC CCTCAAATCC CACTCTGTTG AGACCTCCCC CCGACCCAGA GCAATGACAG 9781 CATCTTTATC ATCTCTGCAT CCCCCAGGGC CATCAGCAGG AGGGAAAGGT TCCCTTCTGC 9841 TTAATTGTCA GACAAGCAGT TGAGTTAAGA AATCTGTGAT TATTGTATTG TTGACTATAC 9901 ACAGCACATT TTAGGGCTCT ATCAAAATAA ATCTGTCCCT TTAAAAAAAG TTAACTAAAG 9961 CCGGGCACGG TGGCTCATGC CTGTAATCCC AACACTTTGG GAGGCTGAGG CAGGCGGATC 10021 CTTGAGCTCA GGAGTTAGAG ACCTGGACTG GGCAAAATGG TGAGGACCCC ATCTCTATAA 10081 AAAATACAAA AATTAGCAAG GTGTGGTAAT GTGCACCAGT GGTCCCAGCT ACTAGAGAGG 10141 CCAAGGTGGG AGGATCATCT GGGCCCGGGG GATGAGGCTG CAGTGAGCCA TGATCGTGCC 10201 ACTGCACTCT AGCCTGGGTA ACAAAGCGAG ACCCTGTCTC TAAATACATC AATCAAATAA 10261 AAATTTTAA AAGTTAA (الرقم التفقي الثلاثي: 64). The amino acid sequence corresponding to the human SARM1 protein can be found on NP_055892.2: 1 MVLTLLLSAY KLCRFFAMSG PRPGAERLAV PGPDGGGGTG PWWAAGGRGP REVSPGAGTE 61 VQDALERALP ELQQALSALK QAGGARAVGA GLAEVFQLVE EAWLLPAVGR EVAQGLCDAI 121 RLDGGLDLLL RLLQAPELET RVQAARLLEQ ILVAENRDRV ARIGLGVILN LAKEREPVEL 181 ARSVAGILEH MFKHSEETCQ RLVAAGGLDA VLYWCRRTDP ALLRHCALAL GNCALHGGQA 241 VQRRMVEKRA AEWLFPLAFS KEDELLLRHA CLAVAVLATN KEVEREVERS GTLALVEPLV 301 ASLDPGRFAR CLVDASDTSQ GRGPDDLQRL VPLLDSNRLE AQCIGAFYLC AEAAIKSLQG 361 KTKVFSDIGA IQSLKRLVSY STNGTKSALA KRALRLLGEE VPRPILPSVP SWKEAEVQTW 421 LQQIGFSKYC ESFREQQVDG DLLLRLTEEE LQTDLGMKSG ITRKRFFREL TELKTFANYS 481 TCDRSNLADW LGSLDPRFRQ YTYGLVSCGL DRSLLHRVSE QQLLEDCGIH LGVHRARILT 541 AAREMLHSPL PCTGGKPSGD TPDVFISYRR NSGSQLASLL KVHLQLHGFS VFIDVEKLEA 601 GKFEDKLIQS VMGARNFVLV LSPGALDKCM QDHDCKDWVH KEIVTALSCG KNIVPIIDGF 661 EWPEPQVLPE DMQAVLTFNG IKWSHEYQEA TIEKIIRFLQ GRSSRDSSAG SDTSLEGAAP 721 MGPT (Serial identification number: 65). As used in this document, the term "SARM1-mediated neurological disease" refers to a neurological disease, disorder, injury, and / or axonal degeneration mediated by SARM1. The term "% sequence match" or "percentage of sequence match" with respect to a reference DNA sequence is defined as the percentage of nucleotides, nucleosides, or nuclear bases in a candidate sequence that are identical to nucleotides, nucleosides, or nuclear bases in the reference DNA sequence, after optimal sequence alignment and the insertion of gaps or spurious sequences, if necessary, to achieve the maximum percentage of sequence match. Alignment for the purpose of determining the percentage of DNA sequence match can be achieved using various methods known in the field, for example, by using publicly available computer programs, such as those described in current protocols in molecular biology (Ausubel). et al . (See Supplement 30, Section 7.7.18, Table 7.7.1), including BLAST, BLAST-2, ALIGN, and Megaalign (DNASTAR), and Clustal W2.0 or Clustal X2.0. Those experienced in the field can determine the appropriate alignment parameters, including any algorithms required to achieve maximum alignment over the full length of the sequences being compared. The percentage of "sequence match" can be determined by comparing two optimally aligned sequences via the comparison window. The portion of the DNA sequence in the comparison window may include additions or deletions (e.g., gaps or spikes) compared to the reference sequence (which does not include additions or deletions) to achieve optimal alignment of the two sequences. The percentage can be calculated by determining the number of positions where the nucleotide, nucleoside, or nuclear base is located in both sequences to produce a number of matching positions, then dividing the number of matching positions by the total number of positions in the comparison window, and then multiplying the result by 100 to obtain the percentage of sequence matching.The output is the percentage of the intended sequence match with respect to the query sequence. The term "polypeptide" or "protein," as used in this document, refers to a polymer of amino acid residues. The term applies to polymers containing naturally occurring amino acids and polymers containing one or more non-naturally occurring amino acids. As used here, the terms "RNAi," "RNAi factor," "iRNA," "iRNA factor," or "RNA interference factor" refer to a factor that mediates the sequence-specific degradation of a target mRNA by RNA interference, for example, via the RNA-derived silencing complex (RISC) pathway. In some applied models, the RNAi factor includes a directional and an antidirectional strand, and the directional and antidirectional strands form a pairing ( example (double-stranded RNA). As used here, "braid" refers to a single, adjacent sequence of nucleotides linked together by internucleotide bonds ( For example (Phosphate diester bonds or phosphorothiolate bonds). The strand can have two free ends ( For example (End 5', and end 3'). As used here, the term "treatment" or "cure" refers to all processes that may slow, control, delay, halt, or alleviate the symptoms of the disorders or diseases identified in this document, but does not necessarily imply the complete elimination of all symptoms of the disorder or disease. Treatment includes administering a protein, nucleic acid, vector, or formula to treat a disease or condition in a patient, particularly in humans. The following examples are given to illustrate inventions, not to restrict them. Examples Example 1: Generation and characterization of receptor-binding proteins TfR Generation of receptor-binding proteins TfR human Antibodies against the human TfR receptor were also generated by immunizing AlivaMab® transgenic mice with the extracellular domains of human transferrin receptor protein 1 (hTfR-ECD-6His, serial number: 67, see Table 8) and mouse transferrin receptor protein 1 (mTfR, serial number: 66). Antigen-positive B cells were sorted from pooled spleens. The binding of individual antibodies cloned from these B cells to the his-labeled hTfR receptor extracellular domain was verified. An additional antibody against the human TfR receptor was generated by immunizing AlivaMab® transgenic mice with the apical domain of human transferrin receptor protein 1 (hTfR-ApD-6His, serial number: 68, see Table 8). Antigen-positive B cells were sorted from pooled spleens. The binding of individual antibodies cloned from these B cells to the his-labeled extracellular domain of the hTfR receptor was verified. Table 8. Immunogen sequences used to generate antibodies to receptors TfR திய்ப்பிர் உுர்பிரை. The mother of immunity Sequence الرقيم التفافي الإشرامي mTfR-ECD-6His HHHHHHCKRVEQKEECVKLAETEETDKSETMETEDVPTSSRLYWADLKTLLSEKLNSIEFADTIKQLSQNTYTPREAGSQKDESLAYYIENQFHEFKFSKVWRDEHYVKIQVKSsIGQNMVTIVQSNGNLDPVESPEGYVAFSKPTEVSGKLVHANFGTKKDFEELSYSV NGSLVIVRAGEITFAEKVANAQSFNAIGVLIYMDKNKFPVVEADLALFGHAHLGTGDPPYTPGFPSFNHTQFPPSQSSGLPNIPVQTISRAAAEKLFGKMEGSCPARWNIDSSKLELSQNQNVKLIVKNVLKERRILNIFGVIKGYEEPDRYVVVGAQRDALGAGVAAKS SVGTGLLLKLAQVFSDMISKDGFRPSRSIIFASWTAGDFGAVGATEWLEGYLSSLHLKAFTYINLDKVVLGTSNFKVSASPLLYTLMGKIMQDVKHPVDGKSLYRDSNWISKVEKLSFDNAAYPFLAYSGIPAVSFCFCEDADYPYLGTRLDTYEALTQKVPQLNQMVRTAAEVAGQLIIKLTHDVELNLDYEMYNSKLLSFMKDLNQFKTDIRDMGLSLQWLYSARGDYFRATSRLTTDFHNAEKTNRFVMREINDRIMKVEYHFLSPYVSPRESPFRHIFWGSGSHTLSALVENLKLRQKNITAFNETLFRNQLALATWTIQGVANALSGDIWNIDNEF 66 hTfR-ECD-6His HHHHHHCKGVEPKTECERLAGTESPVREEPGEDFPAARRLYWDDLKRKLSEKLDSTDFTGTIKLLNENSYVPREAGSQKDENLALYVENQFREFKLSKVWRDQHFVKIQVKDSAQNSVIIVDKNGRLVYLVENPGGYVAYSKAATVTGKLVHANFGTKKDFEDLYTPVNGSIVIVRAGKITFAEKVANAESLNAIGVLIYMDQTKFPIVNAELSFFGHAHLGTGDPYTPGFPSFNHTQFPPSRSSGLPNIPVQTISRAAAEKLFGNMEGDCPSDWKTDSTCRMVTSESKNVKLTVSNVLKEIKILNIFGVIKGFVEPDHYVVVGAQRDAWGPGAAKSGVGTALLLKLAQMFSDMVLKDGFQPSRSIIFASWSAGDFGSVGATEWLEGYLSSLHLKAFTYINLDKAVLGTSNFKVSASPLLYTLIEKTMQNVKHPVTGQFLYQDSNWASKVEKLTLDNAAFPFLAYSGIPAVSFCFCEDTDYPYLGTTMDTYKELIERIPELNKVARAAAEVAGQFVIKLTHDVELNLDYERYNSQLLSFVRDLNQYRADIKEMGLSLQWLYSARGDFFRATSRLTTDFGNAEKTDRFVMKKLNDRVMRVEYHFLSPYVSPKESPFRHVFWGSGSHTLPALLENLKLRKQNNGAFNETLFRNQLALATWTIQGAANALSGDVWDIDNEF 67 hTfR-ApD-6His HHHHHHHHGKPIPNPLLGLDSTGGGGSDSAQNSVIIVDKNGRLVYLVENPGGYVAYSKAATVTGKLVHANFGTKKDFEDLYTPVNGSIVIVRAGKITFAEKVANAESLNAIGVLIYMDQTKFPIVNAELSFFGHAHLGGGGGGLPNIPVQTISRAAAEKLFGNMEGDCPSDWKTDSTCRMVTSESKNVKLTVS 68 Affinity variants were generated from human TfR receptor antibodies by systematically introducing mutations into the individual CDR region of each antibody. The resulting variants were subjected to multiple rounds of selection with decreasing antigen concentrations and / or increasing dissociation times to isolate clones with enhanced affinity rates. Sequences of the individual variants were used to generate a combination sequence set, which underwent an additional round of selection with increasing rigor to identify additional or synergistic mutational pairings between individual CDR regions. The combination clones were then sequenced. Table 1A lists the heavy-chain, light-chain, and VH / VL sequences of the human TfR receptor-binding domains and proteins.Human TfR receptor-binding proteins were generated using recombinant DNA technology. These human TfR receptor-binding proteins can be expressed in a mammalian cell line, such as HEK293 or CHO, and transfected either transiently or stably using an expression system with a predefined optimal heavy-chain:light-chain ratio or a single-chain vector system encoding both the heavy and light chains. The media shown, into which the protein is secreted, can be purified using commonly used techniques. familiarity of connection The binding affinity and chemical valence of human TfR receptor-binding proteins were characterized and measured for the binding of human and long-tailed macaque TFR receptors using a surface plasmon resonance assay on a Biacore 8K instrument equipped with HBS-EP+ buffer (10 mM Hepes pH 7.4 + 150 mM NaCl + 3 mM ETH + 0.05% (w / v) P20 surfactant) at an assay temperature of 37°C. The extracellular domain of the target human and long-tailed macaque TfR receptor was fixed on a CM4 chip (Cytiva P / N 29104989) using standard NHS-EDC amine coupling. TfR receptor-binding proteins were prepared at final concentrations of 0.3, 0.1, 0.033, 0.01, 0.0033, 0.001, 0.00033, and 0.0001 μM respectively by diluting the base solution in the buffer solution used. Linkage analysis was performed using a multi-cycle kinetic approach. Each analysis cycle consisted of (1) injection of the lowest to highest concentration of antibodies onto all Fc cells at a rate of 50 μL / min for 140 seconds, followed by a return to a regulated solution flow for 400 seconds to observe the dissociation phase; (2) replenishment of the slide surfaces by injecting 3 M magnesium chloride for 30 seconds at a rate of 100 μL / min onto all cells; and (3) equilibration of the slide surfaces using a 50 μL (30-second) injection of HBS-EP+. Data were processed using a standard dual reference and prepared for a binding model in State 2 using Biacore 8K Evaluation software to determine the binding rate (k). on M units -1 s -1 and the dissociation rate (k) off s units -1 ), and R max (RU units). The equilibrium dissociation constant (K) is calculated. D From the relationship K D = k off / k on It is measured in molar units. The results are shown in Table 9. Table 9. Binding affinity of receptor-binding proteins TfR The typical human future TfR Human or long-tailed macaque monkey at 37 degrees Celsius Human TfR receptor (TBP) binding proteins K value D For the human TfR receptor (Biacore, nanomolar) At 37 degrees Celsius Standard error of the mean, K value D For the human TfR receptor (Biacore, nanomolar) Number = 3 K value D For the TfR receptor in the long-tailed macaque monkey (Biacore, nanomolar) At 37 degrees Celsius Standard error of the mean, K value D For the TfR receptor in the long-tailed macaque monkey (Biacore, nanomolar) Number = 3 TBP3 32.087 11.795 66.565 11.695 TBP4 153.642 7.949 300.180 2.565 TBP5 0.522 0.284 502.210 8.129 Example 2: Structure and characterization of acids dsRNA Which targets SARM1 In general, single strands (directional and antidirectional) of dsRNA duplicates were synthesized onto a rigid support via MerMade™ 12 (LGC Biosearch Technologies) or a similar automated oligonucleotide synthesizer. In some cases, directional strands were synthesized using a suitable CPG such as 3'-cholesterol-TEG CNA CPG 500 (LGC Biosearch Technologies). The directional and antidirectional strand sequences are shown in Table 5 or 7. Standard reagents were used in the preparation of the oligonucleotides (Table 10), with 0.1 M xanthan hydride in pyridine used as the reagent for sulfur treatment, and 20% diethylamine in acetonitrile used as an auxiliary washing agent after preparation. All monomers (Table 11a) were prepared at 0.1 M in acetonitrile and contained in a molecular sieve trap bag. Oligonucleotides were split and de-protected (C / D) at 45°C for 20 hours. Directional strands were C / D of CPG using ammonia hydroxide (28–30%, cold), while 3% DEA in ammonia hydroxide (28–30%, cold) was used for antidirectional strands. The complete C / D was determined by IP-RP LCMS when the resulting aggregate data confirmed sequence match. Depending on the scale, CPG was filtered through a 0.45 μM PVDF syringe-free filter, a 0.22 μM PVDF Steriflip® vacuum filtration filter, or a 0.22 μM PVDF Stericup® rapid-release filter. The CPG was washed / rinsed again using 30% ACN / RNAse-free water or 30% EtOH / RNAse-free water, then filtered through the same filter and combined with the first filtered mixture. This was repeated twice. The material was then divided equally into 50 mL Falcon tubes for organic removal via Genevac™. After concentration, the crude oligonucleotides were diluted again to the composite scale using RNAse-free water and filtered using a 0.45 µm PVDF syringe-free filter or a 0.45 µm PVDF Steriflip® vacuum filtration.22 micrometers or rapid release of PVDF Stericup® 0.22 micrometers. The crude oligonucleotides were purified using the AKTA™ Pure purification system with anion exchange (AEX) or reverse phase (RP) assays, specifically a 15Q-RP Source column. For AEX, the ES Industry Source™ 15Q column was maintained at 65°C with MPA: 20 mM NaH₂PO₄, 15% ACN, and pH 7.4, and MPB: 20 ​​mM NaH₂PO₄, 1 M NaBr, 15% ACN, and pH 7.4. For RP, the Source™ 15Q-RP column was used with MPA: 50 mM NaOAc with 10% ACN, and MPB: 50 mM NaOAc with 80% ACN. In all cases, fractions with an overall purity greater than 85% and no impurities >5% were combined. The purified oligonucleotides were sweetened using 15 ml of 3K MWCO in centrifuge tubes at 3500 xg For approximately 30 minutes, the oligonucleotides were rinsed with RNAse-free water until the eluent conductivity reached <100 μg / cm. After desalination, 2–3 mL of RNAse-free water was added, and the mixture was aspirated 10x. The retained fraction was transferred to a 50 mL Falcon tube, and this process was repeated until complete transfer of the oligonucleotides was achieved by measuring the concentration of the compound on the filter using a nanodroplet. The final oligonucleotides were then filtered via a 2x nanodroplet through 15 mL of 100K MWCO centrifuge tubes at 3500 rpm. xg For two minutes. The final sweetened oligonucleotides were analyzed for concentration (nanodistillation at A260), characterized by IP-RP LCMS for overall purity and UPLC for UV purity. To prepare the diode, equal quantities of directional and anti-directional braid were combined and heated at 65°C for 10 minutes, then slowly cooled to ambient temperature for over 40 minutes. The integrity of the diode was confirmed by UPLC analysis and characterized by LCMS using IP-RP. All diodes were filtered via nanofiltration, and endotoxin levels were then measured using a Charles River Endosafe® Cartridge to obtain the final RNAi factor compounds. For in vivo analysis, the appropriate amount of diode was dried and then reconstituted in 1X PBS for rodent studies and CSF for non-human primate studies. Table 10 - Reagents for the structure of oligonucleotides Detectors Activator solution (0.5M ETT in ACN) Cap A (Acetic anhydride, pyridine in THF, 1:1:8) Cap B (1-Methylimidazole in THF, 16:84) Oxidation solution (0.02 M iodine in THF / pyridine / water, 70:20:10) Disintegrating solution, 3% TCA in DCM (w / v) Acetonitrile (anhydrous solvent, water maximum 10 ppm) Xanthan hydride (0.1 molar in pyridine) Diethylamine (20% in acetonitrile) Table 11a - Phosphoramide الفوسفور أميديت الاختصارات المورّد رقم الكتالوج CAS DMT-2'-F-A(Bz)-CE الفوسفور أميديت fA هونجين PD1-001 136834-22-5 DMT-2'-F-C(Ac)-CE الفوسفور أميديت fC هونجين PD3-001 159414-99-0 DMT-2'-F-G(iBu)-CE الفوسفور أميديت fG هونجين PD2-002 144089-97-4 DMT-2'-F-U-CE الفوسفور أميديت fU هونجين PD5-001 146954-75-8 DMT-2'-O-Me-A(Bz)-CE الفوسفور أميديت mA هونجين PR1-001 110782-31-5 DMT-2'-O-Me-C(Ac)-CE الفوسفور أميديت mc هونجين PR3-001 199593-09-4 DMT-2'-O-Me-G(iBu)-CE الفوسفور أميديت mG هونجين PR2-002 150780-67-9 DMT-2'-O-Me-U-CE الفوسفور أميديت mU هونجين PR5-001 110764-79-9 5'مكرر(POM) فوسفات الفينيل-2'-Ome-U3'CE فوسفور أميديت POM-VPmU هونجين PR5-032 BVPMUP23B2A1 الجدول 11 ب . بنية متقارن الكوليسترول البنية 1 (متقارن الكوليسترول) المثال 3: توليد عوامل SARM1 RNAi There are specific abbreviations defined as follows: "ACN" stands for acetonitrile; "aAEX" stands for analytic anion exchange; "AS" stands for antisense strand; "DAR" stands for drug / siRNA to antibody / protein ratio; "DCM" stands for dichloromethane; "DHAA" stands for dehydroascorbic acid; "dsRNA" stands for double-stranded ribonucleic acid; "DTT" stands for dithiotriol; "h" stands for hours; "HPLC" stands for ultra-performance liquid chromatography; "LC-MS" stands for liquid chromatography-mass spectrometry; "LTQ / MS" stands for linear ion-absorbing mass spectrometry; "min" stands for minutes; "MW" stands for molecular weight; and "MWCO" stands for the molecular weight limit. The abbreviation "NHS" refers to N-hydroxysuccinimide; the abbreviation "OD" refers to optical density; the abbreviation "PBS" refers to phosphate-buffered saline solution; the abbreviation "PEG" refers to polyethylene glycol; and the abbreviation "RNAi" refers to RNA interference.The abbreviation "rpm" refers to the number of revolutions per minute; the abbreviation "SEC" refers to size exclusion chromatography; the abbreviation "siRNA" refers to small interfering ribonucleic acid; the abbreviation "SMCC" refers to succinimidyl-4-(N-maleidomethyl)cyclohexane-1-carboxylate; the abbreviation "SS" refers to directional strand; the abbreviation "TCO" refers to transfected cyclic octene; the abbreviation "TfR" refers to the transferrin receptor; the abbreviation "THF" refers to tetrahydrofuran; "TRIS" refers to trihydroxymethylaminomethane; the abbreviation "UPLC" refers to ultra-performance liquid chromatography; and the abbreviation "UV" refers to ultraviolet. مخطط التفاعل 1 Q is 1,3,4-oxadiazole or 1 H -Tetrazole. يوضح مخطط التفاعل 1 المسار التخليقي للمركبات الوسيطة المستخدمة لتكوين روابط MSPT وOD النهائية الموضحة في الجدول 4. Reaction diagram 1, step a, depicts the methylation of thiol in compound (1) using iodomethane and a suitable base such as DIEA in a solvent such as THF to obtain compound (2). Step b illustrates the alkylation of compound (2) using ثالثي -Butyl 2-(2-(2-bromoethoxy)ethoxy)acetate is oxidized using a base such as potassium carbonate in a solvent such as acetone to obtain compound (3). Step C shows the oxidation of compound (3) using hydrogen peroxide and ammonium molybdate (VI) tetrahydrate in a solvent such as EtOH, followed by acid deprotection using an acid such as TFA in a solvent such as DCM, to obtain compound (4). It is noted that in the case of 1 H -Tetrazol, deprotection occurred during the oxidation step. Step d shows the coupling reaction of compound (4) with 1-hydroxypyrrolidine-2,5-dione using EDCI in a solvent system such as DCM and THF to obtain compound (5). مخطط التفاعل 2 Reaction diagram 2 shows step A of the coupling of compound (6) with isoindolene-1,3-dione using DIAD and tert-butylphosphine in a solvent such as THF, to obtain compound (7). Step B shows the phosphorylation of compound (7) using 2-cyanoethyl-N,N-diisopropylchlorophosphoramide, in the presence of a base such as DIEA, and in a solvent such as DCM, to obtain compound (8). Preparation process 1 4-(5-(methylthio)- 1H -Tetrazol-1-yl)phenol A solution of 4-(5-mercapto-1H-tetrazol-1-yl)phenol (4.00 g, 20.6 mmol) in THF (50 mL) was cooled to 0°C. DIEA (4.31 g, 33.3 mmol) was added, and the mixture was stirred for 10 minutes before iodomethane (1.54 mL, 24.7 mmol) was added by distillation over 1 minute. The mixture was stirred at 0°C for 20 minutes and then stirred at ambient temperature for 12 hours. After this time, the mixture was diluted with EtOAc (100 mL) and washed with saturated aqueous NH4Cl (2 × 50 mL). The organic layer was separated, dried over sodium sulfate, and concentrated. In the middle of a void To obtain the base compound (4.2 g, 93%). ES / MS Mass / charge: 209 (M+H). The compound listed in Table 12 was prepared in a manner essentially similar to the method used to prepare compound 1. Table 12 Preparing the compound the name Structure ES / MS Mass / charge 2 4-(5-(methylthio)-1,3,4-oxadiazol-2-yl)phenol 209 (M+H) Preparation process 3 Third -Butyl 2-(2-(2-(4-(5-(methylthio)- 1H Tetrazol-1-yl)phenoxy)ethoxy)ethoxy)acetate In a pressure vessel, potassium carbonate (3.15 g, 22.8 mmol) was added to Third -Butyl 2-(2-(2-bromoethoxy)ethoxy)acetate (4.33 g, 14.8 mmol) and 4-(5-(methylthio)-1H-tetrazol-1-yl)phenol (2.5 g, 11.4 mmol) were dissolved in acetone (60 mL). The pressure vessel was sealed and heated at 80 °C for 8 hours with vigorous stirring. After this time, the mixture was cooled to ambient temperature, then filtered by washing with acetone / EtOAc / DCM (30 mL each), and the filtrate was concentrated. In the void Purification was performed using silica gel column chromatography with sequential filtration using 0–100% EtOAc / DCM to obtain the primary compound as a white solid (3.98 g, 85%). ES / MS Mass / charge: 411 (M+H). The compound listed in Table 13 was prepared in a manner essentially similar to the method used to prepare compound 3. Table 13 Preparing the compound the name Structure ES / MS Mass / charge 4 Third -Butyl 2-(2-(2-(4-(5-(methylthio)-1,3,4-oxadiazol-2-yl)phenoxy)ethoxy)ethoxy)acetate 411 (M+H) Preparation process 5 2-(2-(2-(4-(5-(methylsulfonyl)- 1H -Tetrazol-1-yl)phenoxy)ethoxy)ethoxy)acetic acid It was melted Third Butyl 2-(2-(2-(4-(5-(methylthio)-1H-tetrazol-1-yl)phenoxy)ethoxy)ethoxy)acetate (3.98 g, 9.21 mmol) was dissolved in EtOH (100 mL) and cooled to 5–10 °C. Hydrogen peroxide (30%, 19 mL, 184 mmol) was then added, followed by ammonium(VI) molybdate tetrahydrate (1.14 g, 0.921 mmol). The mixture was allowed to rise to ambient temperature, stirred for 4 hours, and then diluted with DCM (150 mL) and washed with a saturated aqueous solution of sodium chloride. The organic phase was separated, dried over sodium sulfate, and concentrated. In the middle of a void The resulting residue was purified by silica gel column chromatography with sequential filtration using 0–100% EtOAc / DCM to obtain the primary compound as a white solid (3.00 g, 80%). ES / MS m / z : 385 (MH). The compound listed in Table 14 was prepared in a manner essentially similar to the method used to prepare compound 5. Table 14 Preparing the compound the name Structure ES / MS Mass / charge 6 Third -Butyl 2-(2-(2-(4-(5-(methylsulfonyl)-1,3,4-oxadiazol-2-yl)phenoxy)ethoxy)ethoxy)acetate 387 (M+H-tBu) It is noted that the conditions were similar, except that the tert-butyl group was not removed during the process. Preparation process 7 2-(2-(2-(4-(5-(methylsulfonyl)-1,3,4-oxadiazol-2-yl)phenoxy)ethoxy)ethoxy)acetic acid TFA (20 mL, 12.0 mmol) was added to a solution consisting of Third Butyl 2-(2-(2-(4-(5-(methylsulfonyl)-1,3,4-oxadiazol-2-yl)phenoxy)ethoxy)ethoxy)acetate (5.60 g, 12.0 mmol) in DCM (60 mL). The mixture was stirred at ambient temperature for 2 hours and concentrated. In the void It was purified by silica gel column chromatography with sequential filtration using 0–100% EtOAc / DCM to obtain the primary compound (4.12 g, 82%). ES / MS m / z : 387 (M+H). Preparation process 8 2,5-Dioxopyrrolidine-1-yl 2-(2-(2-(4-(5-(methylsulfonyl)- 1H Tetrazol-1-yl)phenoxy)ethoxy)ethoxy)acetate EDCI (1.60 g, 10.3 mmol) was added to a solution of 2-(2-(2-(4-(5-(methylsulfonyl)-1H-tetrazol-1-yl)phenoxy)ethoxy)ethoxy)acetic acid (2.80 g, 7.25 mmol) and 1-hydroxypyrrolidine-2,5-dione (1.33 g, 11.6 mmol) in DCM (50 mL) and THF (70 mL). Another 20 mL of DCM was added to dissolve the mixture, and stirring was continued at ambient temperature for 12 hours. The solution was then concentrated. In the void It was purified by silica gel column chromatography with sequential filtration using 0–100% EtOAc / DCM to obtain the primary compound (2.61 g, 65%). ES / MS m / z : 484 (M+H). The compound listed in Table 15 was prepared in a manner essentially similar to the method used to prepare compound 8. Table 15 Preparing the compound the name Structure ES / MS Mass / charge 9 2,5-Dioxopyrrolidine-1-yl-2-(2-(2-(4-(5-(methylsulfonyl)-1,3,4-oxadiazole-2-yl)phenoxy)ethoxy)ethoxy)acetate 484 (M+H) Preparation process 10 2-((( 2R, 3R, 4R, 5R )-5-(2,4-Dioxo-3,4-Dihydroxypyrimidine-1) 2H (-yl)-3-hydroxy-4-methoxytetrahydrofuran-2-yl)methyl)isoindoline-1,3-dione A solution consisting of 1-((2R,3R,4R,5R)-4-hydroxy-5-(hydroxymethyl)-3-methoxytetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione (30 g, 120 mmol), isoindoline-1,3-dione (21 g, 140 mmol), DIAD (27 mL, 140 mmol), tert-butylphosphine (36 mL, 150 mmol), and THF (300 mL) was stirred at ambient temperature for 12 hours. The crude reaction was filtered and concentrated. In the middle of a void It was purified by silica gel flash chromatography with sequential filtration using 0-100% EtOAc / hexane to obtain the basic compound in the form of a white solid (6.0 g, 13%). Preparation process 11 2-Cyanothyl 2R, 3R, 4R, 5R )-5-(2,4-Dioxo-3,4-Dihydroxypyrimidine-1) 2H )-yl)-2-((1,3-Dioxoisoindoline-2-yl)methyl)-4-methoxytetrahydrofuran-3-yl)diisopropylphosphoramide A solution consisting of 2-(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydroxypyrimidine-1(2H)-yl)-3-hydroxy-4-methoxytetrahydrofuran-2-yl)methyl)isoinduline-1,3-dione (3.00 g, 7.74 mmol), 2-cyanoethyl-N,N-diisopropylchlorophosphoramidet (2.47 mL, 11.6 mmol), DIEA (4.05 mL, 23.2 mmol), and DCM (40 mL) was stirred at ambient temperature. After 1 hour, more 2-cyanoethyl-N,N-diisopropylchlorophosphoramidet (0.82 mL, 3.8 mmol) was added. After one hour, the crude mixture was poured into a silica gel slurry (15 g) in 30 ml of 1% TEA / DCM, and concentrated. In the middle of a void It was ground into a dry powder and purified by silica gel flash chromatography with sequential filtration using 40-100% EtOAc / hexanesate (0.5% TEA) to obtain the basic compound in the form of a white foam (3.70 g, 81%). 1 H NMR (d6-DMSO) d 11.4 (br s, 1 H), 7.96-7.78 (m, 5H), 5.83 (dd, 1H), 5.71 (dd, 1H), 4.46-3.47 (m, 9H), 3.39 (s, 1.5H), 3.35 (s, 1.5H), 2.82-2.73(m, 2H), 1.16-0.97(m, 12H). 31 P NMR (d6-DMSO) d 149.7, 149.4. Preparation process 12 Functionally derived directional braid with a tetrazol linker at the end 3’ To a solution of sodium bicarbonate (24 mg, 0.29 mmol) and SARM1-SSC6A (70 mg, 2.41 mL, 4.00 mmol in water), a solution of 2,5-dioxopyrolidine-1-yl 2-(2-(2-(4-(5-(methylsulfonyl)-1H-tetrazol-1-yl)phenoxy)ethoxy)ethoxy)acetate (1.2 mL, 400 mmol in MeCN) was added. An additional 1.2 mL of MeCN was added to adjust the water:MeCN ratio to 1:1, and the mixture was shaken at ambient temperature. After 1.5 hours, LTQ analysis confirmed the reaction was complete. The tube contents were transferred to a 3000 Dalton molecular weight limit (MWCO) membrane centrifuge filter, and the oligonucleotide was rinsed with 10 × 20 mL of RNAse-free water. The retained fraction was collected. The above procedures were repeated in two experiments using 70 and 35 mg SARM1-SSC6A, and the recovered product batches were collected. The final batch volume was 34 mL. The optical density measurement (A260) showed a value of 125.71, corresponding to a concentration of 590 µmol, or 153 mg. The ES / MS result was 7638.5. double series SARM1 Linked The MSPT-SARM1 oriented strand (33 mL, 590 µmol in water) was mixed with the SARM1 counter-oriented strand (7.047 mL, 3.45 mM in water) in a sample tube, and the mixture was shaken for 10 seconds. UPLC analysis confirmed complete annealing. The mixture was filtered using a 100 kDa molecular weight limit (MWCO) membrane centrifuge filter, and the filtrate was lyophilized. The dried powder was reconstituted in 12 mL of water. The resulting solution was analyzed by optical density measurement (A260, 587), which corresponds to a concentration of 1.592 mM, or 292.76 mg. LTQ / MS analysis was also performed at mass / charge ratios: 7686.3, 7638.3. coupling dsRNA With receptor-binding proteins TfR Amino acid residues of native or engineered cysteine ​​were used in TfR receptor-binding proteins to bind dsRNA. Cysteine ​​can be engineered into the basic amino acid sequence of TfR receptor-binding proteins. The method of introducing cysteine ​​as a conjugation medium was described in WO 2018 / 232088, which is incorporated in its entirety for reference and specifically addresses conjugation via cysteine ​​residues. For the binding of engineered cysteine, TfR receptor-binding proteins were first reduced using 40 molar equivalents of dithiothreitol (DTT) at 37°C for 2 hours, followed by desalting to remove the reducing agent using colloidal separation columns or desalting. The TfR receptor-binding protein was then re-oxidized to reconstitute the structural disulfides using 10 molar equivalents of dihydroscorbic acid (DHAA) at ambient temperature for 2 hours. A follow-up desalting process was performed to remove the oxidizing agent. The conjugation of dsRNA with TfR receptor-binding proteins was also performed using the following methods. coupling diagram The conjugation method used 3'SS tetrazole (MSPT) dsRNA, which conjugates to engineered cysteine ​​of TfR receptor-binding proteins. In this method, the TfR receptor-binding protein was prepared as described above to make the engineered thiol available for conjugation by subjecting the TfR receptor-binding proteins to reduction and oxidation. This was followed by overnight incubation of the MSPT-dsRNA with the TfR receptor-binding proteins at 1.2 to 2 molar equivalents for conjugation at ambient temperature. receptor-binding protein conjugation TfR With a link 3’ MSPT Activating the single-chain of RNA pertaining to SARM1 Using a link SMCC To a solution of sodium bicarbonate (178 mg, 2.12 mmol) and SARM1 SS-3C6A (14,000 mL, 5.032 mmol in water), a freshly prepared solution of (2,5-dioxopyrolidine-1-yl)4-[(2,5-dioxopyrol-1-yl)methyl]cyclohexanecarboxylate (14.09 mL, 50 mmol) in acetonitrile was added. The mixture was shaken at ambient temperature for 1.5 hours, and LTQ analysis confirmed the reaction was complete. The mixture was acidified with 100 mL of 0.1 M NaH₂PO₄ (pH 6), filtered through a 0.22 µm filter, and rinsed with water. Salts were removed from the mixture using tangential flow filtration (2 kDa molecular weight limiting membrane, Hydrosart), followed by washing with 2 L of RNAse-free water. The retained fraction was collected, frozen, and freeze-dried. The dried powder was re-soluble in 31 mL of RNAse-free water. The optical density was measured at 463.1, corresponding to a concentration of 2.285 mM and a total mass of 519.27 mg. The extinction coefficient was 202.68, and the LTQ / MS result was 7331.6 (mass / charge). Duplication SMCC-dsRNA Then in a conical tube containing SS-SARM1-AMINO- SMCC with SMCC-Amino-C6 (12.05 mL, 0.016 mmol, 1.328 mmol / L) was supplemented with its analog SS-SARM1-ANTISENSE containing 5'-E-phenylphosphonate (0.0165 mmol, 2.619 mmol / L). The solutions were shaken at 25°C for 30 minutes to obtain the desired SMCC functional group dsRNA (SMCC-dsRNA), then cooled to 10°C for storage. Samples of the heat-treated solutions were analyzed for purity using LTQ and undistorted ultra-performance liquid chromatography (UPLC). Undistorted UPLC analysis (operated at 10°C) showed a single main peak with 92% purity. LTQ-MS: (m / z observed uncoiler of the braid against the trend = 7768.4, and the calculated mass is 7769.04; (m / z observed uncoiler of the braid trend = 7360.4, and the calculated mass is 7361.17). coupling diagram The standard conjugation method used SMCC-mediated dsRNA was employed to conjugate to engineered cysteine ​​of TfR receptor-binding proteins. In this method, the TfR receptor-binding protein was prepared using the same procedure described above to make the engineered thiol available for conjugation by undergoing reduction and oxidation of the TfR receptor-binding proteins. This was followed by incubation of the SMCC-mediated dsRNA with a molar equivalence of 1.2 with the TfR receptor-binding proteins, allowing for overnight conjugation at room temperature. Optionally, after conjugation, a maleimide hydrolysis step can be performed to secure the linker load in the terminal stage and prevent deconjugation during circulation in the human body by adding a back-action Michael reaction. This succinimide ring hydrolysis was carried out by raising the pH of the conjugate to 9.0 using 50 mM arginine (a basic solution of 0.7 mM arginine, pH 9.0, was used) and incubating the solution at 37°C for 20 hours. The maleimide hydrolysis was confirmed by characterizing the +18Da effect by liquid chromatography-mass spectrometry, which occurs upon the addition of water to the succinimide ring. Step 1a: Conjugation of the TfR receptor-binding protein with the SMCC ligand Step 1b: Conjugation of the TfR receptor-binding protein with an SMCC linker loop opening Conjugation was monitored using analytical anion-exchange chromatography (ALCC). A ProPac™ SAX-10 high-performance liquid chromatography column, with a particle size of 10 μm, a diameter of 4 mm, and a length of 250 mm, was used as follows: flow rate 1 mL / min, buffer solution A: 20 mM TRIS at pH 7.0, buffer solution B: 20 ​​mM TRIS at pH 7.0 + 1.5 M NaCl, at 30 °C. The drug / siRNA to antibody / protein ratio (DAR) was calculated based on the peak area ratio from the analytical anion exchange chromatogram (aAEX). After conjugation of dsRNA to the TfR receptor-binding protein, excess dsRNA and unconjugated protein were removed by further purification. Either preparative size exclusion chromatography (SEC) or preparative anion-exchange chromatography was used to purify the final conjugate. Preparative size exclusion chromatography was performed using a Cytiva Superdex® 200 in 1X phosphate-buffered saline at pH 7.2 under isothermal conditions. Alternatively, anion-exchange chromatography, e.g., a ThermoFisher POROS™ XQ, was used with a starting buffer solution of 20 mM TRIS at pH 7.0 and sequential filtration using a 20-column size gradient with a buffer solution containing 20 mM TRIS at pH 7.0 and 1 M NaCl. This resulted in a purified dsRNA conjugate bound to the TfR receptor-binding protein, free of excess dsRNA and a trace amount of unconjugated protein. The properties of the resulting conjugate were analyzed by anionic exchange analytical method to obtain the final DAR quantity (Tables 16). Table 16. Ratio siRNA / medicine to TBP / antibody DAR) Average drug-to-antibody ratio (DAR) DAR0 ratio DAR1 ratio DAR2 ratio TBP5-SMCC-dsRNA No. 8 0.99 1.12 98.06 0.83 TBP5-SMCC-dsRNA No. 7 1 0 100 0 TBP5-SMCC-dsRNA No. 6 1 0 100 0 TBP5-MSPT-dsRNA No. 8 1 0.40 99.6 0 Example 4: Identifying the characteristics of factors SARM1 RNAi In the laboratory SARM1 gene inhibition was assessed In the laboratory Using the TBP5-SMCC-dsRNA 8 conjugate in SH-SY5Y cells, compared with both unbound dsRNA 8 and cholesterol-bound dsRNA 8. cell culture SH-SY5Y and treating the factor RNAi And its analysis: SH-SY5Y cells (ATCC CRL-2266) were derived from the SK-N-SH neuroblastoma cell line (Ross, RA, et al., 1983. J Natl Cancer Inst 71, 741-747). The base medium consisted of a 1:1 mixture of ATCC's Lower Eagle Base Medium (Cat No. 30-2003) and F12 medium. The complete growth medium was supplemented with 10% fetal bovine serum, 1X amino acids, 1X sodium bicarbonate, and 1X penicillin-streptomycin (Gibco). The cells were incubated at 37°C in a humidified atmosphere of 5% CO2. On day 1, SH-SY5Y cells were coated with a fibronectin-coated tissue culture plate with 96 holes and allowed to adhere overnight. On the second day, the complete media was removed and replaced with RNAi factor in serum-free medium. Cells were incubated with RNAi factor for 72 hours, then the medium was changed and RNAi factor was added again for an additional 72 hours, for a total drug incubation time of 144 hours before gene expression analysis. Changes in gene expression in RNAi-treated SH-SY5Y cells were measured using Cells-to-C kits. T Following the manufacturer's protocol (ThermoFisher A35377), pre-designed gene expression assays (available as 20X mixtures) were selected from Applied Biosystems (Foster City, CA, USA). The efficiency of these assays (ThermoFisher Hs00240906_m1 SARM1 and ThermoFisher Hs99999905_m1 GAPDH) was characterized by a cDNA dilution sequence. RT-qPCR was performed on 384-hole MicroAmp Optical reaction plates using a QuantStudio 7 Flex system. The delta-delta CT method was used for normalization to the accumulated GAPDH gene to determine the relative quantification of gene expression. GraphPad Prism v9.0 was used to determine the IC50 using a four-parameter logistic fit. The results are shown in Figure 2 and Table 17. The IC50 activity of the TBP5-SMCC-SARM1 dsRNA conjugate No. 8 was 5.79 nM, a value similar to that of the cholesterol-binding dsRNA conjugate No. 8. The maximum inhibition of the SARM1 gene achieved using the TBP5-SMCC-dsRNA conjugate No. 8 was 56%. Table 17: Factor Activity SARM1 RNAi In the laboratory factor SARM1 RNAi SH-SY5Y, 6d IC50 (nanomolar) SH-SY5Y, 6d Reduction percentage ( KD) SARM1 At a rate of 1 micromolar Unpaired dsRNA No. 8 unavailable 13% TBP5-dsRNA No. 8 5.79 56% Cholesterol-associated dsRNA No. 8 6.39 78% Example 5. In vivo characterization of conjugates TBP-dsRNA Humans in genetically modified mice with the transferrin receptor TfR human ( hTfR ) against SARM1 To determine the efficacy of human transferrin receptor-binding protein (TfR) dsRNA conjugates against the SARM1 gene, they were tested in transgenic mice expressing the human TfR receptor, in which the extracellular domain of the transferrin receptor has been "humanized." TBP5-SMCC-dsRNA conjugate No. 8 was administered intravenously to TfR-expressing transgenic mice at a dose of 10 mg / kg siRNA, either as a single dose or four weekly doses, for comparison with a control group given PBS (n = 5 per group). Twenty-eight days after the first dose, the mice were anesthetized, perfused, and then sacrificed. Hemispheres of the brain were then harvested and processed to assess changes in gene expression using RT-qPCR. Figure 3A illustrates a significant decrease in SARM1 mRNA in both hemispheres of the brain in the TBP5-SMCC-dsRNA #8 dosing groups. A single dose showed a 64% decrease, while four weekly doses showed a 73% decrease compared to the baseline group receiving PBS. Furthermore, assessment of the calf muscle showed no decrease in SARM1 mRNA with a single dose and a 36% decrease with four weekly doses compared to the baseline group receiving PBS (Figure 3B). Taken together, these results indicate that the TBP5-dsRNA #8 conjugate exhibits higher activity in the brain compared to peripheral tissues. Example 6: Determining the properties of conjugates dsRNA receptor-binding proteins TfR human In the living body Following the demonstration of the concept of peripheral siRNA delivery to the central nervous system via the blood-brain barrier in mice, the pharmacokinetic properties of SARM1 siRNA conjugates to human TfR receptor-binding proteins in non-human primates (NHPs) were evaluated as follows. Two- to three-kilogram rhesus monkeys were injected intravenously into the saphenous vein of the thigh with either i) phosphate-buffered saline (n = 4), ii) TBP5-dsRNA #7 (n = 4) at a concentration of 10 mg / kg of active siRNA, or iii) TBP5-dsRNA #6 (n = 4) at a concentration of 10 mg / kg of active siRNA and sacrificed 29 days after the first dose. The anesthetized animals underwent deep cardiac perfusion, and brain, spinal cord, and peripheral tissues were collected. The brain was divided into small sections, and 3 mm of the identified subregions were collected and frozen. Tissues were also collected from the spinal cord, liver, and muscles to assess the levels of mRNA targeted by RT-qPCR in tissue homogenates.Total RNA was isolated from NHP tissue using the RNadvance Tissue Kit (Beckman Coulter, Indianapolis, IN) manually or on a Biomek i7 liquid processor (Beckman Coulter), following the manufacturer's procedures with some modifications. Briefly, frozen tissue sections were mixed with a 5-mm stainless steel ball, lysis buffer, and proteinase K, and homogenized for five 30-second cycles at 1200 rpm, with a 20-second interval between cycles, on a 2010 GenoGrinder (SPEX SamplePrep, Mitochine, NJ). Some tissues were shaved on dry ice prior to homogenization. The homogenates were incubated at 37°C for 1 hour and then extracted with an equal volume of phenol-chloroform. RNA was purified in the supernatant using the RNadvance Tissue Kit, which included a 30-minute digestion process with DNase. The concentration and purity (A260 / A280) of the RNA emulsion were determined by spectroscopy.RNA was normalized to 15 ng / 10 μL by polymerase chain reaction (PCR) and digested again with ezDNase (ds-DNA specific) before reverse transcription using the SSIV VILO kit (Thermo Fisher Scientific, Waltham, MA). Expression of the relevant gene targets in cDNA was determined using TaqMan qPCR assays on the QuantStudio 7 Pro platform (Thermo Fisher Scientific). SARM1 gene expression was titrated using GAPDH or ACTB with their respective probes (Thermo Fisher). The tissues analyzed and their abbreviations include: LDRG, dorsolumbar root ganglion; LSC, thoracic spinal cord; LSC, cervical spinal cord; LSC, lumbar spinal cord; LDRG, dorsolumbar root ganglion; medulla oblongata; pons; midbrain; and MCTX, motor cortex. To determine SARM1 protein levels, approximately 20 mg of frozen sections of neural tissue biopsies were mixed with a chilled RIPA buffer (Pierce, catalog no. 89901, Thermo Scientific) containing protease and phosphatase inhibitors (Halt™ Protease and Phosphatase Inhibitor Cocktail, Thermo Scientific) at a ratio of 20 mL of buffer to 1 g of tissue. The tissue-RIPA mixture was homogenized using a 5 mm stainless steel bead on a 2010 GenoGrinder (Spex SamplePrep). The homogenate was then centrifuged in a refrigerated centrifuge (Eppendorf, Hamburg, Germany), and the supernatant was transferred, divided into several single-use portions, and stored at -80°C for further analysis. The protein concentration in the protein lysate was determined using the Pierce™ BCA Protein Assay Kit (Thermo Scientific), following the manufacturer's instructions. Specifically, the diluted bovine serum albumin parameters were analyzed sequentially in duplicate; each sample of the protein lysate was diluted 10-fold or 20-fold in water and then analyzed individually, respectively. The protein concentration in the undiluted sample was then obtained by calculating the average protein concentration derived from the 10-fold and 20-fold diluted samples. The level of SARM1 protein in the proteolyte was measured using an in-house developed MSD assay. Briefly, a small-format, 96-well MSD GOLD streptavidin SECTOR plate (Meso Scale Diagnostics, Rockville, MD) was simultaneously blocked with bovine serum albumin and coated with a capture antibody (biotin-treated mouse monoclonal antibody against hSarm1, MAB7037, R&D Systems, Minneapolis, MN) and shaken at ambient temperature for 1 hour. After washing, the wells in each plate were incubated with either the proteolyte or recombinant hSARM1 protein (ab271737, Abcam, Waltham, MA) in the presence of MSD Blocker A (Meso Scale Diagnostics), and shaken at ambient temperature for 2 hours. The plates were then washed again, and subsequently incubated with the detection body, a mouse monoclonal antibody against hSARM1 bound to SULFO-TAG (W16079A, BioLegend, San Diego, CA) in the presence of MSD Blocker A with shaking at ambient temperature for 1 hour.After incubation, the plates were washed, and then a 2x MSD-T buffer solution (Meso Scale Diagnostics) was added. The electrochemical fluorescence signal was then measured using an MSD SQ120MM plate reader within 5 minutes of adding the buffer solution. To minimize variability, all biopsies from the same brain region, including those treated with the control and those treated with the test substance, were analyzed, along with a set of serially diluted recombinant hSARM1 protein standards, on the same MSD plate. All samples, including the recombinant protein standards, were analyzed in duplicate. DISCOVERY WORKBENCH (Meso Scale Diagnostics) software was used to analyze the crude electrochemical fluorescence signal and calculate the corresponding protein concentration. Briefly, the crude electrochemical fluorescence signal was first corrected for the background. A standard curve was constructed on each MSD plate by fitting the corrected signal (Y-axis) and the SARM1 protein concentration (X-axis) in each of the serially diluted SARM1 protein standards to a 4-parameter nonlinear logistic regression model. The SARM1 protein concentration in each sample was then calculated inversely from the corresponding corrected signal, based on the standard curve.The SARM1 protein level in each sample was normalized to the total protein level, and the remaining SARM1 protein expression in the treated group was calculated as a percentage of the remaining SARM1 protein expression in the treatment group, relative to the average expression of this protein in the control group treated with the control substance. Administration of the TBP5-SARM1 siRNA conjugate (dsRNA No. 7 in Table 7) via single peripheral intravenous administration at a dose of 10 mg / kg in non-human primates resulted in a significant reduction in SARM1 protein in key brain and spinal cord regions compared to the phosphate-buffered saline (PBS) control group 29 days post-dose. As shown in Figure 4A, a marked reduction in SARM1 protein was observed in the midbrain (60%) and motor cortex (62%). Administration of the TBP5-SARM1 siRNA conjugate (dsRNA No. 6 in Table 7) via single peripheral intravenous administration at a dose of 10 mg / kg in non-human primates resulted in a significant reduction in SARM1 protein in key brain and spinal cord regions compared to the phosphate-buffered saline (PBS) control group 29 days post-dose. As shown in Figure 4B, a significant decrease in SARM1 protein was observed in the thoracic spinal cord (32%), pons (34%), midbrain (60%), and motor cortex (62%). Example 7: Identifying the characteristics of factors SARM1 RNAi In the laboratory SARM1 protein reduction was assessed In the laboratory Using TBP5-SMCC-dsRNA conjugate number 8 or TBP5-MSPT-dsRNA conjugate number 8, in SH-SH5Y cells, the results were compared. cell culture SH-SY5Y and treating the factor RNAi And its analysis: SH-SY5Y cells (ATCC CRL-2266) were derived from the SK-N-SH neuroblastoma cell line (Ross, RA, et al., 1983. J Natl Cancer Inst 71, 741-747). The base medium consisted of a 1:1 mixture of ATCC's Lower Eagle Base Medium (Cat No. 30-2003) and F12 medium. The complete growth medium was supplemented with 10% fetal bovine serum, 1X amino acids, 1X sodium bicarbonate, and 1X penicillin-streptomycin (Gibco). The cells were incubated at 37°C in a humidified atmosphere of 5% CO2. On day 1, SH-SY5Y cells were coated with a fibronectin-coated tissue culture plate with 96 holes and allowed to adhere overnight. On the second day, the complete media was removed and replaced with RNAi factor in serum-free medium. Cells were incubated with RNAi factor for 72 hours, then the medium was changed and RNAi factor was added again for an additional 72 hours, for a total drug incubation time of 144 hours before gene expression analysis. Changes in gene expression in RNAi-treated SH-SY5Y cells were measured using Cells-to-C kits. T Following the manufacturer's protocol (ThermoFisher A35377), pre-designed gene expression assays (available as 20X mixtures) were selected from Applied Biosystems (Foster City, CA, USA). The efficacy of these assays (ThermoFisher Hs00240906_m1 SARM1 and ThermoFisher Hs99999905_m1 GAPDH) was highlighted. Using a cDNA dilution sequence, RT-qPCR was performed on 384-hole MicroAmp Optical reaction plates using a QuantStudio 7 Flex system. The delta-delta CT method was used for normalization to the accumulated GAPDH gene to determine the relative quantification of gene expression. GraphPad Prism v9.0 was used to determine the IC50 using a four-parameter logistic fit. Table 18 presents the results. The IC50 activity of the TBP5-SMCC-SARM1 dsRNA conjugate #8 and the TBP5-MSPT-SARM1 dsRNA conjugate #8 was 0.09 nM and 2.78 nM, respectively. The maximum SARM1 protein inhibition was 41% for the TBP5-SMCC-dsRNA conjugate #8 and 59% for the TBP5-MSPT-SARM1 dsRNA conjugate #8 in SH-SY5Y cells. Table 18: Factor Activity SARM1 RNAi In the laboratory factor SARM1 RNAi SH-SY5Y, 6d IC50 (nanomolar) SH-SY5Y, 6d Reduction percentage ( KD) SARM1 At a rate of 1 micromolar TBP5-SMCC-dsRNA No. 8 0.09 41.03% TBP5-MSPT-dsRNA No. 8 2.78 58.97% Example 8. In vivo characterization of conjugates TBP-SARM1 dsRNA Humanity using a link MSPT In mice genetically modified for the transferrin receptor TfR human ( hTfR) To determine the efficacy of SARM1-specific dsRNA-binding proteins (hTfR) conjugates using an MSPT linker, these were tested in transgenic mice expressing the human TfR receptor, in which the extracellular domain of the transferrin receptor has been "humanized." The TBP5-SMCC-dsRNA conjugate #8 and the TBP5-MSPT-dsRNA conjugate #8 were administered intravenously to hTfR-expressing transgenic mice with a single 10 mg / kg dose of siRNA, for comparison with a PBS-treated control group (n = 3 per group). Twenty-eight days after the first dose, the mice were anesthetized, perfused, and then sacrificed. Hemispheres of the brain were then harvested and processed to assess changes in gene expression using RT-qPCR. Figure 6A illustrates a significant decrease in SARM1 mRNA in both hemispheres of the brain in the TBP5-SMCC-dsRNA #8 and TBP5-MSPT-dsRNA #8 dose groups, with a single dose showing a 55% reduction compared to the baseline PBS-administered group. Furthermore, assessment of the lumbar spinal cord revealed a 52% and 53% decrease in SARM1 mRNA after a single dose compared to baseline in the PBS group, using TBP5-SMCC-dsRNA #8 and TBP5-MSPT-dsRNA #8, respectively (Figure 6B). Overall, these data demonstrate that the TBP5-MSPT-dsRNA #8 conjugate exhibits similar activity in both hemispheres and the lumbar spinal cord compared to TBP5-SMCC-dsRNA #8. List of sequences Serial identification number Sequence 1 SYSMN 2 SISSSSSYIYYADSVKG 3 RHGYSNSDAFDN 4 RASQGISHYLV 5 AASSLQS 6 LQHNSYPWT 7 EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSS 8 DIQMTQSPSAMSASVGDRVTITCRASQGISHYLVWFQQKPGKVPKRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPWTFGQGTKVEIK 9 EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKC 10 DIQMTQSPSAMSASVGDRVTITCRASQGISHYLVWFQQKPGKVPKRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 11 EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSSASTKGPCVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKCDKTHTGGGGQGGGGQGGGGQGGGGQGGGGQEVQLLESGGGLVQPGGSLRLSCAASGRYIDETAVAWFRQAPGKGREFVAGIGGGVDITYYADSVKGRFTISRDNSKNTLYLQMNSLRPEDTAVYYCGARPGRPLITSKVADLYPYWGQGTLVTVSSPP 12 DIQMTQSPSAMSASVGDRVTITCRASQGISHYLVWFQQKPGKVPKRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQCGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 13 EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSSASTKGPXVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG، حيث تكون X عبارة عن S أو C. 14 EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPEVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVSTLPPSQEEMTKNQVSLMCLVYGFYPSDIXVEWESNGQPENNYKTTPPVLDSDGSFFLYSVLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG, where X is A or C. 15 ESKYGPPCPPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQGDMTKNQVQLTCLVKGFYPSDIXVEWESNGQPENNYKTTPPVLDSDGSFFLASRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG, where X is A or C. 16 EVQLVESGGGLVKPGGSLRLSCVASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRHGYSNSDAFDNWGQGTLVTVSSASTKGPCVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVSTLPPSQEEMTKNQVSLMCLVYGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSVLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 17 ESKYGPPCPPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQGDMTKNQVQLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLASRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 18 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSYAIEWVRQAPGQGLEWMGGILPGSGTINYNEKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARMSSNSDQGFDLWGQGTLVTVSSASTKGPXVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG، حيث تكون X عبارة عن S، أو C. 19 DIQMTQSPSSLSASVGDRVTITCKASQGISRFLSWFQQKPGKAPKSLIYAVSSLVDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCVQYNSYPYGFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 20 ETAVA 21 GIGGGVDITYYADSVKG 22 RPGRPLITSKVADLYPY 23 EVQLLESGGGLVQPGGSLRLSCAASGRYIDETAVAWFRQAPGKGREFVAGIGGGVDITYYADSVKGRFTISRDNSKNTLYLQMNSLRPEDTAVYYCGARPGRPLITSKVADLYPYWGQGTLVTVSSPP 24 GGGGQGGGGQGGGGQGGGGQ 25 GSYWIC 26 CIYSTSGGRTYYASWVKG 27 GDDSISDAYFDL 28 QSSQSVYNNNRLA 29 DASTLAS 30 QGTYFSSGWSWA 31 QSLEESGGDLVKPEGSLTLTCTASGFSFSGSYWICWVRQAPGKGLEWIGCIYSTSGGRTYYASWVKGRFTISKTSSTTVTLQMTSLTAADTATYFCARGDDSISDAYFDLWGPGTLVTVSS 32 ALDMTQTASPVSAAVGGTVTINCQSSQSVYNNNRLAWYQQKPGQPPKLLIYDASTLASGVPSRFKGSGSGTQFTLTISGVQSDDSATYYCQGTYFSSGWSWAFGGGTEVVVK 33 QSLEESGGDLVKPEGSLTLTCTASGFSFSGSYWICWVRQAPGKGLEWIGCIYSTSGGRTYYASWVKGRFTISKTSSTTVTLQMTSLTAADTATYFCARGDDSISDAYFDLWGPGTLVTVSSASTKGPCVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 34 ALDMTQTASPVSAAVGGTVTINCQSSQSVYNNNRLAWYQQKPGQPPKLLIYDASTLASGVPSRFKGSGSGTQFTLTISGVQSDDSATYYCQGTYFSSGWSWAFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 35 GUUGCUCGACUCUAACCGCUA 36 UAGCGGUUAGAGUCGAGCAACGG 37 UUCGCCAACUAUUCUACGUGA 38 UCACGUAGAAUAGUUGGCGAAGG 39 ACCUCGCCAACUAUUCUACA 40 UGUAGAAUAGUUGGCGAAGGUCU 41 CCGCAAGAGGUUCUUUAGGGA 42 UCCCUAAAGAACCUCUUGCGGGU 43 mG*mU*mUmGmCmUmCmGfAfCfUmCmUmAmAmCmCmGmC*mU*mA 44 mU*fA*mGmCfGmGmUfUmAmGmAmGmUfCmGfAmGmCmAmAmC*mG*mG 45 mU*mU*mCmGmCmCmAmAfCfUfAmUmUmCmUmAmCmGmU*mG*mA 46 mU*fC*mAmCfGmUmAfGmAmAmUmAmGfUmUfGmGmCmGmAmA*mG*mG 47 mA*mC*mCmUmUmCmGmCfCfAfAmCmUmAmUmUmCmUmA*mC*mA 48 mU*fG*mUmAfGmAmAfUmAmGmUmUmGfGmCfGmAmAmGmGmU*mC*mU 49 mC*mC*mGmCmAmAmGmAfGfGfUmUmCmUmUmUmAmGmG*mG*mA 50 mU*fC*mCmCfUmAmAfAmGmAmAmCmCfUmCfUmUmGmCmGmG*mG*mU 51 mU*fC*mAmCfGmUfAmGmAmAmUmAmGfUmUfGmGmCmGmAmA*mG*mG 52 mU*fC*fAmCmGmUfAmGmAmAmUmAmGfUmUfGmGmCmGmAmA*mG*mG 53 mU*fC*mAmCmGmUmAmGmAmAmUmAmGfUmUmGmGmCmGmAmA*mG*mG 54 mU*fA*mGmCfGmGfUmUmAmGmAmGmUfCmGfAmGmCmAmAmC*mG*mG 55 mU*fA*fGmCmGmGfUmUmAmGmAmGmUfCmGfAmGmCmAmAmC*mG*mG 56 mU*fA*mGmCmGmGmUmUmAmGmAmGmUfCmGmAmGmCmAmAmC*mG*mG 57 mU*fC*mCmCfUmAfAmAmGmAmAmCmCfUmCfUmUmGmCmGmG*mG*mU 58 mU*fC*fCmCmUmAfAmAmGmAmAmCmCfUmCfUmUmGmCmGmG*mG*mU 59 mU*fC*mCmUmAmAmAmGmAmAmCmCfUmCmUmUmGmCmGmG*mG*mU 60 mU*fA*mGmCmGfGmUmUmAmGmAmGmUfCmGfAmGmCmAmAmC*mG*mG 61 mU*fC*mAmCmGfUmAmGmAmAmUmAmGfUmUfGmGmCmGmAmA*mG*mG 62 mU*fG*mUmAmGfAmAmUmGmUmGfGmCfGmAmGmGmU*mC*mU 63 mU*fC*mCmCmUfAmAmAmGmAmCmCfUmCfUmGmCmGmG*mG*mU 64 1 ATCTCCCAGC TCAGCCGAGC CCGTGCCCAG GCCACGCTTT GTTCCAGCCG CCGCCTCCTC 61 TACCCTACGG CGTCCGGAGC CATCCCTCGC CTGCTCGCTC TCTCCTTTCG CCCACTCCCT 121 GCATCTGGGC CTGCATCACC TTTGCCAACC GCTCCCCCGA TCCTGCCGAC ACTCCTCCCC 181 CAAACTTCTG ACCGGCACCC TTGCCTGGTA CCCTTCTCTC CATTCCTCCC CCTCCATCTT 241 CTTTCCCCGA CCCCTCTCGG GTCCCTCTTT TCCCAAAACC CGGGTCTCTC CGCGTGGCCC 301 CGCCTCCAGG CCGGGGATGT CCCCCGCGGC CCCGCGCCCA TGGTCCTGAC GCTGCTTCTC 361 TCCGCCTACA AGCTGTGTCG CTTCTTCGCC ATGTCGGGCC CACGGCCGGG CGCCGAGCGG 421 CTGGCGGTGC CTGGGCCAGA TGGGGGCGGT GGCACGGGCC CATGGTGGGC TGCGGGTGGC 481 CGCGGGCCCC GCGAAGTGTC GCCGGGGGCA GGCACCGAGG TGCAGGACGC CCTGGAGCGC 541 GCGCTGCCGG AGCTGCAGCA GGCCTTGTCC GCGCTGAAGC AGGCGGGCGG CGCGCGGGCC 601 GTGGGCGCCG GCCTGGCCGA GGTCTTCCAA CTGGTGGAGG AGGCCTGGCT GCTGCCGGCC 661 GTGGGCCGCG AGGTAGCCCA GGGTCTGTGC GACGCCATCC GCCTCGATGG CGGCCTCGAC 721 CTGCTGTTGC GGCTGCTGCA GGCGCCGGAG TTGGAGACGC GTGTGCAGGC CGCGCGCCTG 781 CTGGAGCAGA TCCTGGTGGC TGAGAACCGA GACCGCGTGG CGCGCATTGG GCTGGGCGTG 841 ATCCTGAACC TGGCGAAGGA ACGCGAACCC GTAGAGCTGG CGCGGAGCGT GGCAGGCATC 901 TTGGAGCACA TGTTCAAGCA TTCGGAGGAG ACATGCCAGA GGCTGGTGGC GGCCGGCGGC 961 CTGGACGCGG TGCTGTATTG GTGCCGCCGC ACGGACCCCG CGCTGCTGCG CCACTGCGCG 1021 CTGGCGCTGG GCAACTGCGC GCTGCACGGG GGCCAGGCGG TGCAGCGACG CATGGTAGAG 1081 AAGGCGCGCAG CCGAGTGGCT CTTCCCGCTC GCCTTCTCCA GCTGCTTCGG 1141 CTGCACGCCT GCCTCGCAGT AGCGGTGTTG GCGACTAACA AGGAGTGGA GCGCGAGGTG 1201 GAGCGCTCGG GCACGCTGGC GCTCGTGGAG CCGCTTGTGG CCTCGCTGGA CCCTGGCCGC 1261 TTCGCCCGCT GTCTGGTGGA CGCCAGCGAC ACAAGCCAGG GCCGCGGGCC CGACGACCTG 1321 CAGCGCCTCG TGCCGTTGCT CGACTCTAAC CGCTTGGAGG CGCAGTGCAT CGGGGCTTTC 1381 TACCTCTGCG CCGAGGCTGC CATCAAGAGC CTGCAAGGCA AGACCAAGGT GTTCAGCGAC 1441 ATCGGCGCCA TCCAGAGCCT GAAACGCCTG GTTTCCTACT CTACCAATGG CACTAAGTCG 1501 GCGCTGGCCA AGCGCGCGCT GCGCCTGCTG GGCGAGGAGG TGCCACGGCC CATCCTGCCC 1561 TCCGTGCCCA GCTGGAAGGA GGCCGAGGTT CAGACGTGGC TGCAGCAGAT CGGTTTCTCC 1621 AAGTACTGCG AGAGCTTCCG GGAGCAGCCAG GTGGATGGCG ACCTGCTTCT GCGGCTCACG 1681 GAGGAAC TCCAGACCGA CCTGGGCATG AAATCGGGCA TCACCCGCAA GAGGTTCTTT 1741 AGGGAGCTCA CGGAGCTCAA GACCTTCGCC AACTATTCTA CGTGCGACCG CAGCAACCTG 1801 GCGGACTGGC TGGGCAGCCT GGACCCGCGC TTCCGCCAGT ACACCTACGG CCTGGTCAGC 1861 TGCGGCCTGG ACCGCTCCCT GCTGCACCGC GTGTCTGAGC AGCAGCTGCT GGAAGACTGC 1921 GGCATCCACC TGGGCGTGCA CCGCGCCCGC ATCCTCACGG CGGCCAGAGA AATGCTACAC 1981 TCCCCGCTGC CCTGTACTGG TGGCAAACCC AGTGGGGACA CTCCAGATGT CTTCATCAGC 2041 TACCGCCGGA ACTCAGGTTC CCAGCTGGCC AGTCTCCTGA AGGTGCACCT GCAGCTGCAT 2101 GGCTTCAGTG TCTTCATTGA TGTGGAGAAG CTGGAAGCAG GCAAGTTCGA GGACAAACTC 2161 ATCCAGAGTG TCATGGGTGC CCGCAACTTT GTGTTGGTGC TATCACCTGG AGCACTGGAC 2221 AAGTGCATGC AAGACCATGA CTGCAAGGAT TGGGTGCATA AGGATTGT GACTGCTTTA 2281 AGCTGCGGCA AGAACATTGT GCCCATCATT GATGGCTTCG AGTGGCCTGA GCCCCAGGTC 2341 CTGCCTGAGG ACATGCAGGC TGTGCTTACT TTCAACGGTA TCAAGTGGTC CCACGAATAC 2401 CAGGAGGCCA CCATTGAGAA GATCATCCGC TTCCTGCAGG GCCGCTCCTC CCGGGACTCA 2461 TCTGCAGGCT CTGACACCAG TTTGGAGGGT GCTGCACCCA TGGGTCCAAC CTAACCAGTC 2521 CCCAGTTCCC CAGCCCTGCT GTGACTTCCA TTTCCATCGT CCTTTCTGAA GGAACAGCTC 2581 CTGAAACCAG TCTCCCTGGG CTGAGACAAC CTGGGCTCTT CTTAGGAAAT GGCTCTCCCT 2641 CCCCCTGTCC CCCACCCTCA TGGCCCACCT CCAACCCACT TTCCTCAGTA TCTGGAGAGG 2701 GAAGGGAAGT CAGGCTTGGG CACGGGAGGT TAGAACTCCC CCAGGCCCTG CCATTGGGTT 2761 GTCTGTCTCC GTCATGGGGA GGGTCCCTGC TCAGTTCTGG AGACACTGGA GTTGGGGTGG 2821 GGGTGGTTCT GCATTCCCTT CTCCTGCTGA TAGCAGTCAG CTTGAGGAGG ATGACGGAAG 2881 GCAGCCTCAG ACAGGAATTA AGGCAATGCC CAGGCGGGCC TGGGCACTGT ATTCTGAGCA 2941 AGGGCCTGGG CCCAGGAGCC AGCCAGGGAT GAGTGCCATC ATGGCTCTCC ACTCAGACTG 3001 TGCCTGGCCC CTGCACTTAC AACTTCCTGC CGCTCTGTGG CCTTGCCCTG TAATCACTCA 3061 GTGCCCTTAG CTAGCCTGAC TAAGTCCCAG ATCCCCTACA GCTTCCTTCG GTGTGGTATC 3121 TTTTGCCACA TCCAGGGCGA GGGTTGAGGC AAACCAGCCC TCCCTCTGAC TTCCTTGTCA 3181 CTGCAGCCAG CTTTGCTGCA CTTGCTGGTG CACAGGAGCC TCCTGTTTGG GCCTGGGTCT 3241 GGGCATGGGG AGGCCGTGCC TCAAAGCCCA CCCTACCCCA TGCCTTGGTG CTGTGCCTCA 3301 GGCTCCTTCC TGGTCTGGCC CAGCTGGCTT CCCCAGCCCC TCAGCCATCC AGGGCTACCC 3361 ACTGCTTACT CAGGGACCAG GCAGCCCCCA TGGCAGTAAA AGCAGCCTAG ACAGAACCTG 3421 CAGCTCTGTG GAAAGAGGCA AAGTCCTGAA AAGGCAAAGG GTTGTCACTT AGGGCAGCTT 3481 CTCCAACTTT AACATGCATC CAAGTCACCT GGGAATGTTG TTAAAATCAG GAGATCTGGG 3541 GTGGGGCCTA GGACTCTGCA TTTCTTACAG ATTCCCAGGT GAGCTGATGC TGGTGGTTAA 3601 GGGTAGCAAA TCTCTAAAGC ACGAAGCCCT CACAAATCTT TGCCATTTCC CAAACACTCC 3661 GCTCCATGGT CTCCAGTCAT CAGAGCAACT CTACCTGGTA TTATCATCCC CATTTTACAG 3721 ATAATGACAC TGAGGCTCAG AAAGGTTGAG GATAAGCCCA CTTTCCTGTC ATTAGTGGCA 3781 GCCCCAGATC CAGACCTAGG CCTCCTGGCA CCCAGTCCAC TGGCAGTGGA ATTGCTTTCC 3841 TGAGAATCAT TCTGAGGCTG GGCTATTGCT TCTCCCTTGC TTCAAAGAAT CTAGCAGCGG 3901 GGGATAGGAT TTTGCAACAA AAAGCTGACC CAGAGGCCAT ACAGAGCAGG AATATCCCAT 3961 TGCCCCCTCC TCCACTGGGT TCAGAGGGTA AGAAAGCACC CTCCAATAAA CCCAGGCTCC 4021 AGGCCGTGGG GGCTGCTGAA GGCTCTTTCC CCGCAAGGGC CAGGTGTTGA CACCTTAAAG 4081 CTGGCTGCGC CCCCAGCCCC ACTCTTGGCT GTGCTGGCCA GGTGACTCCT AGTTCTTGGC 4141 CACATCATCA GAAAGTCAAA GGTCTCACTC CAGGTTTGGG GCTCCTTCCT TCCACTCCCC 4201 TCCCTGCCAG AGTCTGTCTT GGCCAGTGCC AGCCTCGATG CTTTGGTTTT GACCCCACCT 4261 GATCCTCCTT TCCTCATGCA GCACAAGTGC TCACCGGGGC CAGAGCCAGG GCATGGATAT 4321 GACAAGCAGG GCAGCCTGGA CACTGCCCTC ACAGGACAGC GCCAATAACA ATACAGTGTC 4381 TGAGTATCTC CAGGGGATGA TTTCTGGCTC TTTGTCTCCA ATCAGTCCCA CTCCCTCCTG 4441 AGGTCCCCAA GGGCAGTATT CAGAGAGGTT TCCTGCGTTT TATTTCTATT TGGTATACCC 4501 TCCACTGTTG TCCACTGCCC TGTGTGGCCT TCTGGTTGAC CTCTGCCCGA TCTTCTGTCT 4561 CTCTGAGGGA ATCAGAGTCC AGCATCCAGC CCCAGCTGGA ACAGCTGAAG TCACAAGCCT 4621 CCTCTAAGCC AAGGCCAGTG TGTTCAGAGG TGACTGCCAC CCATACTAGG ACAAACACAG 4681 CTCAGATCAC CAGGTCAAGC ACCTAGGCCT GGCTTCTCCT GAGACAGAGG ACTCAGAAGT 4741 GGCCTTTCCT CCAAAGCCTG CTCAGACACA GGTCTGTAGG GCCAGGGTGT TCTGCTTGGC 4801 TGGGCTGCAG CTGCTACCCC TCGGTTGGGG CTGAGTCAGC CAGATCCTCC CCCTACTTCT 4861 CCCCAAGGGC CAAGAACTGC TCAGGGACAT TAAAGGTCAA AAGTCCAGCC ACACTCATTC 4921 ATCCTTTCCC CAGGCCCATG AAGAGAGGCA TCTCATTGTA GAATGTATGA GGAAGTGGGGA 4981 AGTATCTCAG AGAATCAGCT AAGTTTCCTA ACTTGTCCAT CCAAATGTGA TCACCACGAT 5041 TCAACAATTT GGGGCATTGC TGATCTAGCC GTTCCTAGTG GGGCTTGCTC AAGGTTGCAC 5101 AGCGAGTCAG TAGAAGCCCT GGCTGGCCCC ACTTGGTACC AATCCACCAG GCAGCTCAGG 5161 GCTCCTGCCC AGCCCAGCAG CTTCTGTTGT CTAACGTATG GCAGGCAGAC TGGGAGCAGG 5221 AAAACAGAGG GCCCCAAAGC CCAAGGCACC AGAAGGTTTG TTTCAGTTTG CTGAAGCTGA 5281 TTTGTAATGA TTGGCACTCT TCAGCCAGGG GAGTGGGTAG GCCATAGCCA AGGATCGATT 5341 CCCCAACCAC AGCAAAGGCA ACACTCTTCC TCCAGAGATC ACCAAGCCCC TCTTACCTCC 5401 CTCCCTCCTT CCCAAGGCTG GCACTAACCA GGTACCACAT TCATTGTTAA GGAATGGCTG 5461 ATGACTGCTA CACGTGTTGG GAACCTGGTT GGGGCTGTGC AGTTTGGGCT GGAAGGAGAG 5521 ATGCCAGCCC TCGTGCTGCC TCTGGTCCCT GAAGTGTCAC CTCTCTCAGG ACCTCTCCTC 5581 TGGCCTGTGGG GGTTATAAGT GATGGATAGC AGAAAGGGAG AACTGACTCC TGTCCCAAAT 5641 AGCTCCTCTG CCACCTGTCC TGCAGTGGGC CTGTGTGGGT TATGATTCTA GATCCTAGAC 5701 AGAGGCTGGG TCAGCTGTGG ATGGGGTGGT GCCTTGGTCT CTCTTGACTA CCTCGTCCAA 5761 AGAGAGCACT GCCCTTAGAC AAGAGTTGCT TGTCCTGCTG TGGGCTGGGC TTCCAGCTGC 5821 AGACCTCCAG TTGCTTGGTG TTCACTTTGC TCCTCTTGCC CTCTGTCTTC TGGTCCAGGC 5881 AGATCAGGGG CTCTGGGGAA ACTGCTGGAA CTCGAGGTGA GGATCAGCCT TTTCCAGCAT 5941 CCTGTGAGAG ACCAGAGAGA GAGTTTGGAT TTCATGTGGG GAACCCTCAA GGCCTGTCTG 6001 GAGAAGTGAC ACAGGATTTA CTGGGGTGGG CTGGTCCAGG TAGCTCTCCT GAACCTCCTC 6061 CTTCCCCAAG CTGAGAAGCT GAGAGCTGGA GGACAATATC CAGGGACATG GCTCTGGAAA 6121 ATAACTTTTT TTTTTTTAAG AGACAGGGTC TTGCTCTGTT GTCCAGGCTG GAGGGCAGTG 6181 ACATAATCAT AGCTCACTGT ACCCTTGAAC TCCTGGGCTC AAGTGATCCT CCTGCCTCAG 6241 CCTCCTTAGT AGCTGGGACT ACCAGTGCAT ACCACCATGC CTGGGTGATT TTTTAAATTT 6301 TTTATACAGA CAAGGTCTTG CTATGTTGCC CAGGCTGATC TTGAATTCCC GGGCTCAAGT 6361 GGTCCTCCTG CCTCAGCCTC CCACAGGATC GGGATTACAG GCAAGAGCCT CCACGCCCGG 6421 CCATGAAATA TAATTCTTAA TATCATACAG GAAAAAGTCA GCGGGTCAAG CTAGCCTGTG 6481 GCCCAGCCAC AACTAGCTGA CAAAGCTTCC TGGCCTTCCC TTTAACACAG TTCTGCTGCC 6541 ATAGTTCCAT CTATAAAATG GGAATGGAGG GAAATAGGGG AACTGGGAGA GAGAACACAG 6601 CCTTGCCAAG CAGCAATGTT AGCCTGATCC TTCCTCCACC TAGCTCGCCA TCTCGCCCTT 6661 GGAAAATGGC TCCTGGAGGA TTAGGCAGCC ATCTGCAAGG AGAGGGGCAA CCTGGGACAA 6721 GACACCCAGA GGGTAAGGAT TCCAGGAATG AAGCTGCCAT TTCTGGTTGG GAGGAGAGA 6781 GGAAACTTTT AAGAAAGG GCTCCATTAT GAGCATGGGT TCAGGGCCCT GCATTACCCA 6841 ATCAGAACAG CCGGGATGAG CAGGAGGCCA GCTCCCAGGA GGAAGGGGAA CCCTTCATA 6901 AAGTTCAGAG TGGCTGGGTA GAGTGAGTTG AAGATGCCGG AGGCCGTCAG CATGGCCAGG 6961 CTATTCACAC AGGCCACAGC AGAAAAAGAGA GCACCTGTGA AGAAATAAAT ACCATACTCT 7021 GGAGTCCGAA AGGGCCATAT TCCAACTCTG GCACCACCAC CTCACAGCTG TGTGACCGGG 7081 AGTAGTCACT TAACCTATGT CTCCCCTTCC TCACCAGTAA ATCCTGCTAC ATCATGTACT 7141 GTGACAAGGA TTCAGTAAGG TCATATGTGG ACAGTAGCTG GCACAGAGGG GCTACTAAAC 7201 AAATGGCTGC TATTAAATCC ACATTAAAAG TACATGTGAT CTHEADAC CCHEAD 7261 AAAGAAAAAAA AAAGTACATG TGATATTGTC TGATGAAAGC TTGATGGAAA TGGCTTTTTT 7321 CTGGTTTATC CTCTTTGGAA TCATTCCTG TTTGGGATTA ACTGCTGGTC TGATCAGTTC 7381 CAATATTCAT AGCGGTGTCA CCACTGAATA GCTTCTTATC CTTTGGGTTC CTGTTCCTCC 7441 TTCTGCTAAA TAAGGATAAT ACCTATTTCC TAGATTGTGA GCAAATTAA GTTCACATGG 7501 AAATCACCCA TCACTGGGCC TGGTCCCCTG GAAGTAGCTA GTTAGTAAGG GCTGTTCTTT 7561 TCTCCTGTTT CTCTTGACAT CTCTGGGCAC AGAGAAAGTG CTGGGAAAAA AAGTTTAGGT 7621 GAATGAATGA AGACACATGG ATTCTGGGGA CACCAGAACC CACAGTGGGC TCTGTATGGC 7681 ACCAGAGTCT CTGTCATCAT CAGATCCTCA TTCCAGGACA GATGGAAAAA GATGAATGTT 7741 TCCAGACTGG GGCATAAAGA CCCAGAGGCT GGAGAAGCTG TTCTTTATAG ATATACCCAGG 7801 AGAACCCACA GTTTACAAAA TGTGCAACAA CCCAACAGAA GTTGAGATTA AATTCTGTCA 7861 CATCTAGAGG GGTCTGTGAT GTCATCAAAA GCAAACCACC CACATCACAG ATGAAGAAAC 7921 AGGCCTGTGG CAGGGCTCGG ACTAAAACCC AGATCCTGAG ACCAGCTGCT TTTAAACCA 7981 GACGTAGGTT TGCATCCTAG CTCCACCATT TACTGAGTAA CCTTGGGTGA GCCAATGTAA 8041 CCCCCTGGGT CTCTGTTTCT TTATCTGTCA ACTGTGGAAA ATGAAACCCA TGTCACAAGG 8101 TTGTTCACTT CTGGGCTTGT ACACGCTGAC CCCAGAGAAA CAGGGAACTC TGGCATCACC 8161 ACACCCATCT TACAGACGGA AAAGCTGAG TCTGCAGAGA GTAAATCCTC TGCTCTGGTT 8221 ATCTAGAAAG AACATAATTG TGCTCTGCTG ACTGCAAATC CCAACTCTGC GGTTTGAAAA 8281 TCCAAGGTGG CATGATCCTC TGCCCATTGT GGGCAATTTC ACAGAAATGT GTTTGTTTTG 8341 GCCACTTACT TCTCCAGGGT GAGAGGGGGG AAGGCAAGCT GTTCCCCCAG CCATGGCTGC 8401 CCATCAGCCC GTTTCGGGCA GCACTGGACA TGAGGAACCA GACACAGGTG GGTTCTGACA 8461 CTCACCCTGC TCTGTCTCTC TCACCAGCTT GGAGAGTTTA GCCCGGATGA CAGGTGTGAT 8521 ERROR RESPONSE ERROR ACCCATATCC TGTGERROR RESPONSE TCRESPONSE 8581 GGGGCTGGGG AGAGGGGCGT CCAAGGGAAA GGCAGCAGAG CTCCTATCCA TACCCCACGT 8641 GGGGCTTAGG TTAGACCCAG GAAGAACTTC CTTGATGGTG AGGGTGGGAA GACAGTAGTC 8701 AAGGAGGAAT GGAGACTGCC CTTGTCTGGG CTTGGCCACC TGCTAGCTCT CATGAATGAA 8761 TGCTAATTTCC CATTGATTGC TTTCTTGTCT GAACCTCTTG TGGTCACAGC AGGCATCACC 8821 CACCACTTG GCACTTAGTA GGGATATGGC AGGGCACAGA AAACAAGCAT GGGCTTTGGA 8881 GTCAGCCCTG AGTTCAAAAC CTGATGCCAT TACATATTAT CTGTGTGGCC TGGGGTACTT 8941 ACCCTCTCTG ATCCTGACTC CCTGTATGAG GAAGATAATA AGGCCTTCAT CACAGGATGG 9001 TTCTGAGGCA TAGGAGGCTG AATAATGGTG CCCAATGGCA TCAGATTCAT AGCCCTGGAA 9061 CCTGTAAATA CTACCTTATT TGGAAAATGA GTCTATGCAG GTGTGCAGTT AAGCCTCCTG 9121 CONFUSION GTTATCCTGG ATTAGGTTGG GCCCTAAATG CCGTCACACA TATCTTTATA 9181 AGAGGAAAGC AGACGGAGAT TTGGCACCGA AGATTGAG AAGGCACAAA GAGGAGGAGA 9241 GTCAATGTGA GCACAGAGGC AGAGACTGGT GATGGCCGCC CCAAGCCAAG GAATGCCAGC 9301 AGCCCCAGAA GCTGGAAAA ATGAAACA CGTTCTCTCC TGGAGGCTTG CAUGHT 9361 CTGCCTGCTG ACTGCTTCCA TTCAGCCCGG TGGTACTGAC TTTGGACTTC TGGCCTCCAG 9421 AACTGTGAGA GAATATGTTT CTGTTGTGTT AAGCCCCAA GTTTGTGGTA TGTCATTACA 9481 GCAATCTCAG GGAACCATA ASSISTANCE AAAGGTAACA TCTATGAAGA GCATGGCATA 9541 GGGACACAGC AAATGGGAGT TCCTTTTCCC TTTGCATTCA GTTACTTACA GGCTTCCTGT 9601 TTTCTTCATA ACCATTTCTC TCCCTGTGCG ACTGCTGACT CCTCAGCAAA ACTGCAAACT 9661 CCTACAGGAC AGTGGATCCT CCAAAGAAGG TATACGATGA GGCATCCAGG GACCCTAGCA 9721 GTGTCAGGCC CCTCAAATCC CACTCTGTTG AGACCTCCCC CCGACCCAGA GCAATGACAG 9781 CATCTTTATC ATCTCTGCAT CCCCCAGGGC CATCAGCAGG AGGGAAAGGT TCCCTTCTGC 9841 TTAATTGTCA GACAAGCAGT TGAGTTAAG AATCTGTGAT TATTGTATTG TTGACTATAC 9901 CHALLENGE TTAGGGCTCT ATCAAAATAA ATCTGTCCCT TTAAAAAAG TTAACTAAG 9961 CCGGGCACGG TGGCTCATGC CTGTAATCCC AACACTTTGG GAGGCTGAGG CAGGCGGATC 10021 CTTGAGCTCA GGAGTTAGAG ACCTGGACTG GGCAAAATGG TGAGGACCCC ATCTCTATAA 10081 AAAATACAAA AATTAGCAAG GTGTGGTAAT GTGCACCAGT GGTCCCAGCT ACTAGAGAGG 10141 CCAAGGTGGG AGGATCATCT GGGCCCGGGG GATGAGGCTG CAGTGAGCCA TGATCGTGCC 10201 ACTGCACTCT AGCCTGGGTA ACAAAGCGAG ACCCTGTCTC TAAATACATC AATCAAATAA 10261 AAATTTAAA AAGTTAA 65 1 MVLTLLLSAY KLCRFFAMSG PRPGAERLAV PGPDGGGGTG PWWAAGGRGP REVSPGAGTE 61 VQDALERALP ELQQALSALK QAGGARAVGA GLAEVFQLVE EAWLLPAVGR EVAQGLCDAI 121 RLDGGLDLLL RLLQAPELET RVQAARLLEQ ILVAENRDRV ARIGLGVILN LAKEREPVEL 181 ARSVAGILEH MFKHSEETCQ RLVAAGGLDA VLYWCRRTDP ALLRHCALAL GNCALHGGQA 241 VQRRMVEKRA AEWLFPLAFS KEDELLRLHA CLAVAVLATN KEVEREVERS GTLALVEPLV 301 ASLDPGRFAR CLVDASDTSQ GRGPDDLQRL VPLLDSNRLE AQCIGAFYLC AEAAIKSLQG 361 KTKVFSDIGA IQSLKRLVSY STNGTKSALA KRALRLLGEE VPRPILPSVP SWKEAEVQTW 421 LQQIGFSKYC ESFREQQVDG DLLLRLTEEE LQTDLGMKSG ITRKRFFREL TELKTFANYS 481 TCDRSNLADW LGSLDPRFRQ YTYGLVSCGL DRSLLHRVSE QQLLEDCGIH LGVHRARILT 541 AAREMLHSPL PCTGGKPSGD TPDVFISYRR NSGSQLASLL KVHLQLHGFS VFIDVEKLEA 601 GKFEDKLIQS VMGARNFVLV LSPGALDKCM QDHDCKDWVH KEIVTALSCG KNIVPIIDGF 661 EWPEPQVLPE DMQAVLTFNG IKWSHEYQEA TIEKIIRFLQ GRSSRDSSAG SDTSLEGAAP 721 MGPT 66 HHHHHHCKRVEQKEECVKLAETEETDKSETMETEDVPTSSRLYWADLKTLLSEKLNSIEFADTIKQLSQNTYTPREAGSQKDESLAYYIENQFHEFKFSKVWRDEHYVKIQVKSSIGQNMVTIVQSNGNLDPVESPEGYVAFSKPTEVSGKLVHANFGTKKDFEELSYSVNGSLVIVRAGEITFAEKVANAQSFNAIGVLIYMDKNKFPVVEADLALFGHAHLGTGDPYTPGFPSFNHTQFPPSQSSGLPNIPVQTISRAAAEKLFGKMEGSCPARWNIDSSCKLELSQNQNVKLIVKNVLKERRILNIFGVIKGYEEPDRYVVVGAQRDALGAGVAAKSSVGTGLLLKLAQVFSDMISKDGFRPSRSIIFASWTAGDFGAVGATEWLEGYLSSLHLKAFTYINLDKVVLGTSNFKVSASPLLYTLMGKIMQDVKHPVDGKSLYRDSNWISKVEKLSFDNAAYPFLAYSGIPAVSFCFCEDADYPYLGTRLDTYEALTQKVPQLNQMVRTAAEVAGQLIIKLTHDVELNLDYEMYNSKLLSFMKDLNQFKTDIRDMGLSLQWLYSARGDYFRATSRLTTDFHNAEKTNRFVMREINDRIMKVEYHFLSPYVSPRESPFRHIFWGSGSHTLSALVENLKLRQKNITAFNETLFRNQLALATWTIQGVANALSGDIWNIDNEF 67 HHHHHHCKGVEPKTECERLAGTESPVREEPGEDFPAARRLYWDDLKRKLSEKLDSTDFTGTIKLLNENSYVPREAGSQKDENLALYVENQFREFKLSKVWRDQHFVKIQVKDSAQNSVIIVDKNGRLVYLVENPGGYVAYSKAATVTGKLVHANFGTKKDFEDLYTPVNGSIVIVRAGKITFAEKVANAESLNAIGVLIYMDQTKFPIVNAELSFFGHAHLGTGDPYTPGFPSFNHTQFPPSRSSGLPNIPVQTISRAAAEKLFGNMEGDCPSDWKTDSTCRMVTSESKNVKLTVSNVLKEIKILNIFGVIKGFVEPDHYVVVGAQRDAWGPGAAKSGVGTALLLKLAQMFSDMVLKDGFQPSRSIIFASWSAGDFGSVGATEWLEGYLSSLHLKAFTYINLDKAVLGTSNFKVSASPLLYTLIEKTMQNVKHPVTGQFLYQDSNWASKVEKLTLDNAAFPFLAYSGIPAVSFCFCEDTDYPYLGTTMDTYKELIERIPELNKVARAAAEVAGQFVIKLTHDVELNLDYERYNSQLLSFVRDLNQYRADIKEMGLSLQWLYSARGDFFRATSRLTTDFGNAEKTDRFVMKKLNDRVMRVEYHFLSPYVSPKESPFRHVFWGSGSHTLPALLENLKLRKQNNGAFNETLFRNQLALATWTIQGAANALSGDVWDIDNEF 68 HHHHHHHHGKPIPNPLLGLDSTGGGGSDSAQNSVIIVDKNGRLVYLVENPGGYVAYSKAATVTGKLVHANFGTKKDFEDLYTPVNGSIVIVRAGKITFAEKVANAESLNAIGVLIYMDQTKFPIVNAELSFFGHAHLGGGGGGLPNIPVQTISRAAAEKLFGNMEGDCPSDWKTDSTCRMVTSESKNVKLTVS

Claims

1. A SARM1 RNAi agent comprising Formula (I): (R-L)n-P,wherein R is a double stranded RNA (dsRNA) comprising a sense stand and an antisense strand, wherein the antisense strand is complementary to SARM1 mRNA; wherein L is a linker, or optionally absent; andwherein P is a protein comprising one monovalent human TfR binding domain, wherein the human TfR binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein HCDR1 comprises SEQ ID NO: 1, HCDR2 comprises SEQ ID NO: 2, HCDR3 comprises SEQ ID NO: 3, LCDR1 comprises SEQ ID NO: 4, LCDR2 comprises SEQ ID NO: 5, and LCDR3 comprises SEQ ID NO: 6; and wherein n is an integer of 1 to 3.

2. The SARM1 RNAi agent of claim 1, wherein:(a) n is 1; or(b) n is 2.

3. The SARM1 RNAi agent of claim 1 or 2, wherein VH comprises SEQ ID NO: 7 and VL comprises SEQ ID NO:

8.

4. The SARM1 RNAi agent of any one of claims 1-3, wherein the human TfR binding domain is a Fab, scFv, Fv, or scFab.

5. The SARM1 RNAi agent of any one of claims 1-4, wherein the human TfR binding domain further comprises a heavy chain constant region comprising cysteine at residue 124 (according to the EU Index numbering).

6. The SARM1 RNAi agent of any one of claims 1-5, wherein P further comprises a half-life extender. 7. The SARM1 RNAi agent of claim 6, wherein the half-life extender is an immunoglobulin Fc region, optionally wherein the immunoglobulin Fc region is a modified human IgG4 Fc region.

8. The SARM1 RNAi agent of claim 7, wherein the modified human IgG4 Fc region comprises proline at residue 228, and alanine at residues 234 and 235 (all residues are numbered according to the EU Index numbering).

9. The SARM1 RNAi agent of claim 7 or 8, wherein P comprises an immunoglobulin Fc region comprising cysteine at residue 378 (according to the EU Index numbering).

10. The SARM1 RNAi agent of any one of claims 7-9, wherein the immunoglobulin Fc region comprises:(a) a first Fc CH3 domain comprising a serine at position 349, a methionine at position 366, a tyrosine at position 370, and a valine at position 409; and a second Fc CH3 domain comprising a glycine at position 356, an aspartic acid at position 357, a glutamine at position 364, and an alanine at position 407 (all residues are numbered according to the EU Index numbering); or (b) a first Fc CH3 domain comprising leucine at residue 405, and a second Fc CH3 domain comprising arginine at residue 409 (all residues are numbered according to the EU Index numbering).

11. The SARM1 RNAi agent of any one of claims 1-6, wherein P comprises one heavy chain (HC) and one light chain (LC), wherein HC comprises SEQ ID NO: 9 and LC comprises SEQ ID NO:

10.

12. The SARM1 RNAi agent of any one of claims 1-11, wherein P comprises two heavy chains HC1 and HC2 and one light chain LC1, wherein :(a) HC1 comprises SEQ ID NO: 14, LC1 comprises SEQ ID NO: 10, HC2 comprises SEQ ID NO: 15; or(b) HC1 comprises SEQ ID NO: 16, LC1 comprises SEQ ID NO: 10, HC2 comprises SEQ ID NO:

17.

13. The SARM1 RNAi agent of claim 6, wherein the half-life extender is a VHH that binds HSA.

14. The SARM1 RNAi agent of claim 13, wherein the VHH comprises CDR1 comprising SEQ ID NO: 20, CDR2 comprising SEQ ID NO: 21, and CDR3 comprising SEQ ID NO:

22.

15. The SARM1 RNAi agent of claim 13 or 14, wherein the VHH comprises SEQ ID NO:

23.

16. The SARM1 RNAi agent of any one of claims 13-15, wherein P comprises one heavy chain (HC) and one light chain (LC), and wherein the HC comprises SEQ ID NO: 11 and the LC comprises SEQ ID NO: 12 or 10.

17. The SARM1 RNAi agent of any one of claims 1-10, wherein P is a heterodimeric antibody that comprises a first arm comprising one monovalent human TfR binding domain and a second arm that is a null arm.

18. The SARM1 RNAi agent of claim 17, wherein the second arm comprises one heavy chain (HC) and one light chain (LC), and wherein the HC comprises SEQ ID NO: 18 and the LC comprises SEQ ID NO:

19.

19. The SARM1 RNAi agent of claim 17 or 18, wherein P comprises two heavy chains HC1 and HC2 and two light chains LC1 and LC2, wherein HC1 comprises SEQ ID NO: 13, LC1 comprises SEQ ID NO: 10, HC2 comprises SEQ ID NO: 18, and LC2 comprises SEQ ID NO: 19.

20. The SARM1 RNAi agent of any one of claims 1-19, wherein L is a Mal-Tet-TCO linker, SMCC linker, GDM linker, MSPT linker or OD linker.

21. The SARM1 RNAi agent of any one of claims 1-20, wherein L is a SMCC or MSPT linker.

22. The SARM1 RNAi agent of any one of claims 1-21, wherein P is linked to the 3’ end of the sense strand of dsRNA, optionally via the linker.

23. The SARM1 RNAi agent of any one of claims 1-22, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from: (a) the sense strand comprises SEQ ID NO: 35, and the antisense strand comprises SEQ ID NO: 36;(b) the sense strand comprises SEQ ID NO: 37, and the antisense strand comprises SEQ ID NO: 38;(c) the sense strand comprises SEQ ID NO: 39, and the antisense strand comprises SEQ ID NO: 40; and(d) the sense strand comprises SEQ ID NO: 41, and the antisense strand comprises SEQ ID NO: 42,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.

24. The SARM1 RNAi agent of any one of claims 1-23, wherein one or more nucleotides of the sense strand or the antisense strand are modified nucleotides.

25. The SARM1 RNAi agent of claim 24, wherein each nucleotide of the sense strand is a modified nucleotide.

26. The SARM1 RNAi agent of claim 24, wherein each nucleotide of the antisense strand is a modified nucleotide.

27. The SARM1 RNAi agent of any one of claims 24-26, wherein the modified nucleotide is a 2'-fluoro modified nucleotide, 2'-O-methyl modified nucleotide, 2’ deoxy nucleotide (DNA), or 2'-O-C16 alkyl modified nucleotide.

28. The SARM1 RNAi agent of any one of claims 24-27, wherein the sense strand has: (a) four 2'-fluoro modified nucleotides at positions 7, 9, 10, and 11 from the 5’ end of the sense strand; or (b) three 2'-fluoro modified nucleotides at positions 9, 10, and 11 from the 5’ end of the sense strand.

29. The SARM1 RNAi agent of any one of claims 24-28, wherein the antisense strand has: (a) four 2'-fluoro modified nucleotides at positions 2, 6, 14, and 16 from the 5’ end of the antisense strand; (b) five 2'-fluoro modified nucleotides at positions 2, 5, 7, 14, and 16 from the 5’ end of the antisense strand; (c) five 2'-fluoro modified nucleotides at positions 2, 5, 8, 14, and 16 from the 5’ end of the antisense strand; (d) five 2'-fluoro modified nucleotides at positions 2, 3, 7, 14, and 16 from the 5’ end of the antisense strand; or (e) three 2'-fluoro modified nucleotides at positions 2, 14, and 16 from the 5’ end of the antisense strand.

30. The SARM1 RNAi agent of claim 28 or 29, wherein nucleotides at positions other than the listed positions are 2'-O-methyl modified nucleotides.

31. The SARM1 RNAi agent of any one of claims 1-30, wherein the sense strand and the antisense strand have one or more modified internucleotide linkages, optionally wherein the modified internucleotide linkage is phosphorothioate linkage.

32. The SARM1 RNAi agent of claim 31, wherein:(a) the sense strand has four or five phosphorothioate linkages; and / or(b) the antisense strand has four or five phosphorothioate linkages.

33. The SARM1 RNAi agent of any one of claims 1-32, wherein the antisense strand has a phosphate analog at the 5’ end, optionally wherein the phosphate analog is 5’-vinylphosphonate.

34. The SARM1 RNAi agent of any one of claims 1-33, wherein the sense strand comprises an abasic moiety or inverted abasic moiety.

35. The SARM1 RNAi agent of any one of claims 1-34, wherein the sense strand and the antisense strand comprise a pair of nucleic acid sequences selected from: (a) the sense strand comprises SEQ ID NO: 43, and the antisense strand comprises SEQ ID NO: 44, 54, 55, 56, or 60;(b) the sense strand comprises SEQ ID NO: 45, and the antisense strand comprises SEQ ID NO: 46, 51, 52, 53, or 61;(c) the sense strand comprises SEQ ID NO: 47, and the antisense strand comprises SEQ ID NO: 48 or 62; and(d) the sense strand comprises SEQ ID NO: 49, and the antisense strand comprises SEQ ID NO: 50, 57, 58, 59, or 63.

36. The SARM1 RNAi agent of any one of claims 1-35, wherein the sense strand and the antisense strand have a pair of nucleic acid sequences selected from: (a) the sense strand consists of SEQ ID NO: 43, and the antisense strand consists of SEQ ID NO: 44, 54, 55, 56, or 60;(b) the sense strand consists of SEQ ID NO: 45, and the antisense strand consists of SEQ ID NO: 46, 51, 52, 53, or 61;(c) the sense strand consists of SEQ ID NO: 47, and the antisense strand consists of SEQ ID NO: 48 or 62; and(d) the sense strand consists of SEQ ID NO: 49, and the antisense strand consists of SEQ ID NO: 50, 57, 58, 59, or 63.

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

38. The SARM1 RNAi agent of any one of claims 1-36, or the pharmaceutical composition of claim 37, for use in a therapy.

39. The SARM1 RNAi agent of any one of claims 1-36, or the pharmaceutical composition of claim 37, for use in the treatment of a SARM1-mediated neurological disease.

40. The SARM1 RNAi agent or pharmaceutical composition for use of claim 39, wherein the SARM1-mediated neurological 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’s 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, periventricular leukomalacia, a hereditary ataxia, noise-induced hearing loss, congenital hearing loss, age-related hearing loss, Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, Lewy Body Dementia, frontotemporal dementia, tauopathy, synucleinopathy, amyloidosis, diabetic neuropathy, globoid cell leukodystrophy (Krabbe’s disease), Bassen-Komzweig syndrome, transverse myelitis, motor neuron disease, spinocerebellar ataxia, pre-eclampsia, hereditary spastic paraplegias, spastic paraparesis, familial spastic paraplegia, French settlement disease, Strumpell-Lorrain disease, non-alcoholic steatohepatitis (NASH), adrenomyeloneuropathy, progressive supra nuclear palsy (PSP), Friedrich’s ataxia, spinal cord injury, acute optic neuropathy (AON), a genetic or idiopathic retinal condition, 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, an outer retinal neuropathy, optic nerve neuritis, optic nerve degeneration associated with multiple sclerosis, Kjer’s optic neuropathy, ischemic optic neuropathy, chemotherapy-induced peripheral neuropathy, neuromyelitis optica, Charcot Marie Tooth disease, deficiency in vitamin B12, deficiency in folic acid (vitamin B9), isolated vitamin E deficiency syndrome, non-arteritic 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).

41. Use of the SARM1 RNAi agent of any one of claims 1-36 in the manufacture of a medicament for treating a SARM1-mediated neurological disease.

42. The use of claim 41, wherein the SARM1-mediated neurological 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’s 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, periventricular leukomalacia, a hereditary ataxia, noise-induced hearing loss, congenital hearing loss, age-related hearing loss, Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, Lewy Body Dementia, frontotemporal dementia, tauopathy, synucleinopathy, amyloidosis, diabetic neuropathy, globoid cell leukodystrophy (Krabbe’s disease), Bassen-Komzweig syndrome, transverse myelitis, motor neuron disease, spinocerebellar ataxia, pre-eclampsia, hereditary spastic paraplegias, spastic paraparesis, familial spastic paraplegia, French settlement disease, Strumpell-Lorrain disease, non-alcoholic steatohepatitis (NASH), adrenomyeloneuropathy, progressive supra nuclear palsy (PSP), Friedrich’s ataxia, spinal cord injury, acute optic neuropathy (AON), a genetic or idiopathic retinal condition, 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, an outer retinal neuropathy, optic nerve neuritis, optic nerve degeneration associated with multiple sclerosis, Kjer’s optic neuropathy, ischemic optic neuropathy, chemotherapy-induced peripheral neuropathy, neuromyelitis optica, Charcot Marie Tooth disease, deficiency in vitamin B12, deficiency in folic acid (vitamin B9), isolated vitamin E deficiency syndrome, non-arteritic 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).  

Images