Oligonucleotide composition and method thereof

Abstract

The present invention provides oligonucleotides, compositions, and methods for preventing and / or treating various pathological conditions, disorders, or diseases. [Solution] In some embodiments, the provided technology comprises nucleic acid base modification, sugar modification, internucleotide linkage modification and / or patterns thereof, and has improved properties, activity and / or selectivity. In some embodiments, the provided technology targets MAPT. In some embodiments, the disclosure provides MAPT oligonucleotides, compositions and methods for preventing and / or treating MAPT-related conditions, disorders or diseases, such as Alzheimer's disease (AD) or frontotemporal dementia (FTD).

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 62 / 983,742, filed on 1 March 2020, and U.S. Provisional Patent Application No. 63 / 111,071, filed on 8 November 2020, each of which is incorporated herein by reference in its entirety.

[0002] Technical field In particular, this disclosure provides oligonucleotides, compositions, and methods (e.g., preparation, use, etc.) thereof. In some embodiments, the technologies provided are useful for the prevention and / or treatment of various pathological conditions, disorders, or diseases, including various neurodegenerative disorders. [Background technology]

[0003] background Oligonucleotides are useful in a variety of applications, such as therapy, diagnosis, and / or research. For example, oligonucleotides that target various genes may be useful in treating conditions, disorders, or diseases related to such target genes. [Overview of the project] [Means for solving the problem]

[0004] overview In some embodiments, the Disclosure provides MAPT oligonucleotides and compositions thereof with significantly improved properties and / or high activity. In particular, the Disclosure provides techniques for designing, manufacturing and utilizing such oligonucleotides and compositions. In particular, in some embodiments, the Disclosure provides oligonucleotides comprising useful internucleotide binding patterns and / or glycosylation patterns that, when combined with one or more other structural elements, such as nucleotide sequences (or portions thereof), nucleic acid base modifications (and patterns thereof), additional chemical parts, etc., can provide MAPT oligonucleotides and compositions thereof with high activity and / or desirable properties, including effective and efficient reduction of the expression, level and / or activity of MAPT transcripts and the products encoded thereby. In some embodiments, MAPT oligonucleotides and compositions reduce the level of MAPT transcripts and are useful for the treatment and / or prevention of MAPT-related conditions, disorders or diseases, such as Alzheimer's disease (AD) and frontotemporal dementia (FTD).

[0005] In some embodiments, MAPT oligonucleotides have the ability to mediate a knockdown of MAPT such that the level, expression, and / or activity of MAPT or its products is reduced. In some embodiments, MAPT oligonucleotides have the ability to mediate a panspecific knockdown of MAPT such that the level, expression, and / or activity of several or all MAPT alleles is reduced. In some embodiments, MAPT oligonucleotides have a nucleotide sequence complementary to a sequence common to several or all MAPT alleles.

[0006] In some embodiments, the base sequence of the MAPT plasmid is ACGTTGCAGTGTTCCACUAU, ACTATCCTCCTTCAGCUCCU, AGTGTTCCACTATCCUCCUU, ATCCTCCTTCAGCTCCUGCA, CACGTTGCAGTGTTCCACUA, CACTATCCTCCTTCAGCUCC, CAGTGTTCCACTATCCUCCU, CCACGTTGCAGTGTTCCACU, CCACTATCCTCCTTCAGCUC, CGTTGCAGTGTTCCACUA UC, CTATCCTCCTTCAGCUCCUG, GCAGTGTTCCACTATCCUCC, GTGTTTCCACTATCCTCCUUC, GTGTTCCACTATCCTCCUUCAG, GUGUUCCACTATCCTCCTTC, TATCCTCCTTCAGCTCCUGC, TCCACTATCCTCCTTCAGCU, TGCAGTGTTCCACTAUCCUC, TGTTCCACTATCCTCCUUCA, TTCCACTATCCTCCTUCAGC, or TTGCAGTGTTCCACTAUCCU (or in the formula, each T may be independently substituted with U, and vice versa), or containing therein, or containing a span thereof (e.g., 10 to 20 adjacent bases, i.e., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20). In some embodiments, the base sequence of a MAPT oligonucleotide is such a sequence.

[0007] In particular, this disclosure demonstrates that controlling structural elements of MAPT oligonucleotides can significantly affect oligonucleotide properties and / or activity, including knockdown of MAPT (or its products) (e.g., reduction in activity, expression, and / or levels). In some embodiments, MAPT knockdown is mediated by steric hindrance affecting RNase H and / or translation, and / or interference with mRNA maturation. In some embodiments, MAPT knockdown is mediated by mechanisms involving RNA interference or splicing regulation. In some embodiments, MAPT knockdown may occur through multiple mechanisms. In some embodiments, controlled structural elements of oligonucleotides include, but are not limited to, nucleotide sequence, chemical modifications (e.g., modifications of sugars, bases, and / or internucleotide bonds) or their patterns, stereochemistry (e.g., stereochemistry of binding phosphorus in chiral internucleotide bonds) or changes in their patterns, the structure of a first or second wing or core, and / or conjugation with additional chemical moieties (e.g., carbohydrate moieties, targeting moieties, etc.). In particular, in some embodiments, this disclosure demonstrates that the properties and / or activity of MAPT oligonucleotides can be greatly improved by controlling the stereochemistry of the skeletal chiral center (stereochemistry of the bound phosphorus), in conjunction with optionally controlling other aspects of oligonucleotide design.

[0008] In some embodiments, the present disclosure relates to any MAPT oligonucleotides that function through any mechanism and include any sequence, structure, or format (or part thereof) described herein, comprising at least one non-natural modification of a base, sugar, or internucleotide bond.

[0009] In some embodiments, an oligonucleotide composition comprises multiple oligonucleotides, where each oligonucleotide comprises at least one chiral-controlled internucleotide bond, e.g., an internucleotide bond having a conjugate phosphorus that is either Rp or Sp configuration, or is highly purified, rather than a random mixture of Rp and Sp (e.g., about 70–100% (e.g., about 85–100%, 90–100%, 95–100%, or about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) of all oligonucleotides having the same base sequence in the composition share the same stereochemistry with the conjugate phosphorus). In some embodiments, such internucleotide bonds are also referred to as “stereodefined (or stereocontrolled or chiral-controlled) internucleotide bonds.” In some embodiments, such an oligonucleotide composition is referred to as “stereodefined (or stereocontrolled or chiral-controlled) oligonucleotide composition.” In some embodiments, at least one internucleotide bond is a chiral-controlled internucleotide bond and is Sp. In some embodiments, at least one internucleotide bond is a chiral-controlled internucleotide bond and is Rp. In some embodiments, at least 1 to 20 internucleotides, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 internucleotides Each rheotide bond is independently a chiral-controlled internucleotide bond. In some embodiments, each phosphorothioate internucleotide bond is independently a chiral-controlled internucleotide bond. In some embodiments, each internucleotide bond containing chiral phosphorus is independently a chiral-controlled internucleotide bond.

[0010] In some embodiments, the Disclosure provides an oligonucleotide whose nucleotide sequence comprises at least 10 adjacent nucleotides of a sequence identical or complementary to the nucleotide sequence of a MAPT gene or its transcript, the oligonucleotide comprises at least one modified internucleotide bond (an internucleotide bond other than a native phosphate bond, which may exist in various salt forms), and the oligonucleotide has the ability to reduce the level, expression, and / or activity of a MAPT target gene or its gene product.

[0011] In some embodiments, the disclosure relates to a MAPT oligonucleotide composition, the MAPT oligonucleotide comprising at least one chiral internucleotide bond that is not chiralized (for example, the internucleotide bond is sterically random). In some embodiments, the chiralized internucleotide bond is a phosphorothioate internucleotide bond. In some embodiments, the chiralized internucleotide bond is a non-negatively charged internucleotide bond, e.g., n001.

[0012] In some embodiments, the MAPT oligonucleotide includes a non-negatively charged or neutral internucleotide bond.

[0013] In some embodiments, the provided oligonucleotide includes additional chemical moieties such as a carbohydrate moiety or a targeting moiety. In some embodiments, when such moieties are incorporated into the oligonucleotide, one or more properties and / or activities may be improved.

[0014] In some embodiments, this disclosure 1) Common base sequence; 2) Common skeletal connection patterns; and 3) Common skeletal chiral center pattern The present invention provides a chiral-controlled MAPT oligonucleotide composition comprising multiple oligonucleotides sharing a common base sequence, a common skeletal bonding pattern, and a common skeletal chiral center pattern, which is substantially a pure formulation of a single oligonucleotide in that the non-random or controlled levels of oligonucleotides in the composition have a common base sequence, a common skeletal bonding pattern, and a common skeletal chiral center pattern. In some embodiments, the multiple oligonucleotides include a common sugar modification pattern, a common base modification pattern (if any), and a common additional chemical moiety (if any).

[0015] In some embodiments, the MAPT oligonucleotide composition is a chiral-controlled oligonucleotide composition comprising multiple oligonucleotides of a specific oligonucleotide type, which is chiral-controlled in that the oligonucleotides of the specific oligonucleotide type are highly purified compared to a substantially racemic formulation of oligonucleotides having the same nucleotide sequence. In some embodiments, the MAPT oligonucleotide (e.g., a MAPT having a nucleotide sequence that is, contains, or includes at least 15 adjacent nucleotide spans thereof) is sterically random (or not chiral-controlled).

[0016] In some embodiments, the provided oligonucleotide comprises one or more blocks. In some embodiments, a block comprises one or more consecutive nucleosides and / or nucleotides and / or sugars or bases and / or internucleotide bonds that share common chemistry not present in adjacent blocks (e.g., sugars, bases or at least one common modification of an internucleotide bond, or a combination or pattern thereof, or a stereochemical pattern), or vice versa. In some embodiments, a block is a wing or a core.

[0017] In some embodiments, an oligonucleotide comprises two or more components, for example, at least one wing and at least one core. In some embodiments, the wing is structurally different from the core in that it contains structures not present in the core [e.g., stereochemistry, or chemical modifications (or patterns thereof) in sugar, base, or internucleotide bonds], and vice versa. In some embodiments, the structure of an oligonucleotide includes a wing-core-wing structure. In some embodiments, the structure of an oligonucleotide includes a wing-core-wing structure, where one wing is structurally different from the other wing and core [e.g., stereochemistry, additional chemical parts, or chemical modifications (or patterns thereof) in sugar, base, or internucleotide bonds] (e.g., an asymmetric oligonucleotide). In some embodiments, the structure of an oligonucleotide has or includes a wing-core, core-wing, or wing-core-wing structure, where the block is either a wing or a core. In some embodiments, the core is referred to as a gap.

[0018] In some embodiments, the wing includes a sugar modification or pattern thereof that is not present in the core. In some embodiments, the wing includes a sugar modification that is not present in the core. In some embodiments, each sugar in the wing is the same. In some embodiments, at least one sugar in the wing is different from another sugar in the wing. In some embodiments, one or more sugar modifications and / or sugar modification patterns in the first wing of the oligonucleotide (e.g., in the 5'-wing) are different from one or more sugar modifications and / or sugar modification patterns in the second wing of the oligonucleotide (e.g., in the 3'-wing). In some embodiments, the modification is a 2'-OR modification, where R is as described herein. In some embodiments, R is optionally substituted with C 1~4It is alkyl. In some embodiments, the modification is 2'-OMe. In some embodiments, the modification is 2'-MOE. In some embodiments, the modified sugar is a high-affinity sugar, such as a bicyclic sugar (e.g., LNA sugar), 2'-MOE, etc. In some embodiments, the 5'-wing includes a 2-MOE modification. In some embodiments, each 5'-wing sugar is 2'-MOE modified. In some embodiments, the 3'-wing includes a 2-OMe modification. In some embodiments, each 3'-wing sugar is 2'-OMe modified.

[0019] In some embodiments, the internucleotide bond connecting the wing nucleoside and the core nucleoside is considered part of the core. In some embodiments, the internucleotide bond connecting the wing nucleoside and the core nucleoside is considered part of the wing.

[0020] In some embodiments, the wing comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) non-negatively charged internucleotide bonds. In some embodiments, the non-negatively charged internucleotide bonds are neutral internucleotide bonds. In some embodiments, as demonstrated herein, oligonucleotides comprising wings containing non-negatively charged internucleotide bonds can result in high activity and / or selectivity.

[0021] In some embodiments, the core sugar is a natural DNA sugar that does not contain a substitution at the 2' position (the 2'-carbon has two -H atoms). In some embodiments, each core sugar is a natural DNA sugar that does not contain a substitution at the 2' position (the 2'-carbon has two -H atoms).

[0022] In some embodiments, MAPT oligonucleotides or MAPT oligonucleotide compositions are useful for the prevention or treatment of MAPT-related conditions, disorders, or diseases in subjects requiring them. In some embodiments, the Disclosure provides a method for preventing or treating a MAPT-related condition, disorder, or disease, comprising administering to a subject suffering from the condition or a subject susceptible to the condition a therapeutically effective amount of an oligonucleotide or a pharmaceutical composition that can deliver or contain a therapeutically effective amount of an oligonucleotide. In some embodiments, the Disclosure provides a pharmaceutical composition comprising a MAPT oligonucleotide and a pharmaceutically acceptable carrier. In some embodiments, the oligonucleotide in the pharmaceutical composition is in one or more pharmaceutically acceptable salt forms, such as a sodium salt form, an ammonium salt form, etc.

[0023] In some embodiments, oligonucleotides or oligonucleotide compositions are useful in the manufacture of pharmaceuticals for the prevention or treatment of MAPT-related conditions, disorders, or diseases, such as Alzheimer's disease (AD) and frontotemporal dementia (FTD), for subjects requiring them.

[0024] The provided technologies (e.g., oligonucleotides, compositions, methods, etc.) can be used to prevent and / or treat various MAPT-related conditions, disorders, or diseases. In some embodiments, the condition, disorder, or disease is Alzheimer's disease (AD). In some embodiments, the condition, disorder, or disease is frontotemporal dementia (FTD). [Modes for carrying out the invention]

[0025] Detailed description of a specific embodiment The technology of this disclosure can be more readily understood by referring to the following detailed description of specific embodiments.

[0026] definition As used herein, unless otherwise specified, the following definitions shall apply. For the purposes of this disclosure, chemical elements are identified according to the periodic table, CAS versions, and Handbook of Chemistry and Physics, 75th Ed. In addition, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999 and “March's Advanced Organic Chemistry”, 5th Ed., Ed.: Smith, M. b. and March, J., John Wiley & Sons, New York: 2001.

[0027] When used herein, unless otherwise clearly indicated in the context, (i) the terms “a” or “an” may be understood to mean “at least one.” (ii) The term “or” may be understood to mean “and / or”; (iii) The terms “include,” “contain,” “inclusive” (whether used with “not limited to these”) and “inclusive” (whether used with “not limited to these”) may be understood to include the listed components or steps, whether presented alone or with one or more additional components or steps; (iv) The term “another” may be understood to mean at least one or more additional / secondary components; (v) The terms “about” and “approximately” may be understood to allow a standard deviation as a person skilled in the art would understand; and (vi) where a range is provided, the endpoints are included.

[0028] Unless otherwise specified, oligonucleotides and their elements (e.g., their base sequence, sugars) Descriptions of modifications, internucleotide bonds, linked phosphorus stereochemistry, patterns, etc. are given in 5' to 3'. As those skilled in the art will understand, in some embodiments, oligonucleotides may be provided and / or used in salt form, particularly in pharmaceutically acceptable salt form, e.g., sodium salt. Unless otherwise specified, oligonucleotides include various forms of oligonucleotides. As those skilled in the art will also understand, in some embodiments, individual oligonucleotides in a composition may be considered to have the same chemical composition and / or structure, even if a particular oligonucleotide in such a composition (e.g., a liquid composition) could be in one or more different salt forms at a particular moment (and even if it could be dissolved, the oligonucleotide chain may exist in anionic form when in a liquid composition, for example). For example, those skilled in the art will understand that, at a given pH, individual internucleotide bonds along an oligonucleotide chain may be in acidic (H) form or one of several possible salt forms (e.g., sodium salt, or a salt of another cation depending on which ions may be present in the formulation or composition), and that in acidic form (e.g., all cations, if present, H) + You will understand that, insofar as the replacements (by) have the same chemical composition and / or structure, such individual oligonucleotides can be appropriately considered to have the same chemical composition and / or structure.

[0029] Analogues: The term “analogue” includes any chemical moiety that is structurally different from a reference chemical moiety or reference class chemical moiety, but is capable of performing at least one function of such reference chemical moiety or reference class chemical moiety. Non-limiting examples include nucleotide analogues, which are structurally different from nucleotides but perform at least one function of nucleotides; and nucleic acid base analogues, which are structurally different from nucleic acid bases but perform at least one function of nucleic acid bases.

[0030] Antisense: As used herein, the term “antisense” refers to an oligonucleotide or other nucleic acid feature having a nucleotide sequence complementary or substantially complementary to a target nucleic acid that has hybridizing ability. In some embodiments, the target nucleic acid is the target gene mRNA. In some embodiments, hybridization is required, or causes, a decrease in the level, expression, or activity of the target nucleic acid or its gene product in a single activity, for example. The term “antisense oligonucleotide” as used herein refers to an oligonucleotide complementary to the target nucleic acid. In some embodiments, an antisense oligonucleotide has the ability to induce a decrease in the level, expression, or activity of the target nucleic acid or its product. In some embodiments, an antisense oligonucleotide has the ability to induce a decrease in the level, expression, or activity of the target nucleic acid or its product via a mechanism involving RNase H, steric hindrance, and / or RNA interference.

[0031] Chiral Control: As used herein, “chiral control” refers to the control of the stereochemical designation of the chiral bound phosphorus in a chiral internucleotide bond within an oligonucleotide. As used herein, a chiral internucleotide bond is an internucleotide bond in which the bound phosphorus is chiral. In some embodiments, control is achieved via the absence of chiral elements from the sugar and base moieties of the oligonucleotide, for example, in some embodiments, control is achieved via one or more chiral auxiliary groups during the oligonucleotide preparation process, as described herein, and these chiral auxiliary groups are often part of the chiral phosphoramidites used in the oligonucleotide preparation process. In contrast to chiral control, those skilled in the art will understand that conventional oligonucleotide synthesis without chiral auxiliary groups does not allow control of the stereochemistry at the chiral internucleotide bond when forming the chiral internucleotide bond using such conventional oligonucleotide synthesis. In some embodiments, the stereochemical designation of each chiral bound phosphorus in each chiral internucleotide bond within an oligonucleotide is controlled.

[0032] Chiral-controlled oligonucleotide composition: The terms “chiral-controlled oligonucleotide composition,” “chiral-controlled nucleic acid composition,” etc., as used herein, refer to a composition comprising multiple oligonucleotides (or nucleic acids) that share 1) a common base sequence, 2) a common skeletal linkage pattern, and 3) a common skeletal phosphorus modification pattern, wherein the multiple oligonucleotides (or nucleic acids) share the same linkage phosphorus stereochemistry in one or more chiral internucleotide bonds (chiral-controlled or sterically defined internucleotide bonds, where the chiral linkage phosphorus in the composition is Rp or Sp ("sterically defined"), and not a random mixture of Rp and Sp like a non-chiral-controlled internucleotide bond). The levels of the multiple oligonucleotides (or nucleic acids) in the chiral-controlled oligonucleotide composition are predetermined / controlled (e.g., through chiral-controlled oligonucleotide preparation for stereoselective formation of one or more chiral internucleotide bonds). In some embodiments, about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 20%, 30%) of all oligonucleotides in a chiral-controlled oligonucleotide composition. 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) are oligonucleotides among these multiple groups.In some embodiments, about 0.1% to 100% (e.g., 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%) of all oligonucleotides in a chiral controlled oligonucleotide composition sharing a common base sequence. 0%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) are multiple oligonucleotides. In some embodiments, the level is about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 8%) of all oligonucleotides in a composition sharing a common base sequence (e.g., multiple oligonucleotides or oligonucleotide types), or about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30%, 40%, 50%, 60%, 70%, 8%) of all oligonucleotides in a composition sharing a common base sequence, common skeletal linkage pattern, and common skeletal phosphorus modification pattern, or about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30%, 40%, 50%, 60%, 70%, 8%) of all oligonucleotides in a composition sharing the same chemical composition. The percentages are 0%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, multiple oligonucleotides have the same stereochemistry with approximately 1 to 50 chiral internucleotide bonds.In some embodiments, multiple oligonucleotides have the same stereochemistry in approximately 1% to 100% of the chiral internucleotide bond. In some embodiments, several oligonucleotides (or nucleic acids) have the same chemical composition (as those skilled in the art will understand, in some embodiments they may exist in one or more forms, e.g., acid form, salt form, etc.). In some embodiments, several of these oligonucleotides... The nucleotide (or nucleic acid) level is approximately 1% to 100% of all oligonucleotides (or nucleic acids) in the composition that share the same chemical structure as one of the oligonucleotides (or nucleic acids). In some embodiments, each chiral internucleotide bond is a chiral-controlled internucleotide bond, and the composition is a fully chiral-controlled oligonucleotide composition. In some embodiments, the oligonucleotides (or nucleic acids) are structurally identical. In some embodiments, the chiral-controlled internucleotide bonds have a diastereoplecy of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%, typically at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%. In some embodiments, the chiral-controlled internucleotide bonds have a diastereoplecy of at least 95%. In some embodiments, the chiral-controlled internucleotide bond has at least 96% diastereopurity. In some embodiments, the chiral-controlled internucleotide bond has at least 97% diastereopurity. In some embodiments, the chiral-controlled internucleotide bond has at least 98% diastereopurity. In some embodiments, the chiral-controlled internucleotide bond has at least 99% diastereopurity. In some embodiments, the percentage (e.g., levels described herein) is (DS) nc is or at least (DS) ncIn the formula, DS is the diastereopurity as described herein (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% or more), and nc is the number of chiral-controlled internucleotide bonds as described herein (e.g., 1-50, 1-40, 1-30, 1-25, 1-20, 5-50, 5-40, 5-30, 5-25, 5-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more). In some embodiments, the percentage (e.g., the levels described herein) is (DS) nc is or at least (DS) nc In this formula, DS is between 95% and 100%. For example, when DS is 99% and nc is 10, the percentage is 90%, or at least 90% ((99%)). 10≈0.90 = 90%). In some embodiments, the level of multiple oligonucleotides in a composition is expressed as the product of the diastereopurities of each chiral-controlled internucleotide bond in the oligonucleotide. In some embodiments, the diastereopurity of an internucleotide bond linking two nucleosides in an oligonucleotide (or nucleic acid) is expressed by the diastereopurity of an internucleotide bond of a dimer linking the same two nucleosides, where the dimer is prepared under equivalent conditions, and in some examples, the same synthetic cycle conditions (for example, for a bond between Nx and Ny in an oligonucleotide....NxNy....., the dimer is NxNy). In some embodiments, not all chiral internucleotide bonds are chiral-controlled internucleotide bonds, and the composition is a partially chiral-controlled oligonucleotide composition. In some embodiments, the uncontrolled chiral internucleotide bonds have diastereopurities of about 80%, 75%, 70%, 65%, 60%, less than 55%, or about 50%, as is typically observed in sterically random oligonucleotide compositions (e.g., conventional oligonucleotide synthesis, e.g., by phosphoramidite methods, as those skilled in the art will understand). In some embodiments, several oligonucleotides (or nucleic acids) are of the same type. In some embodiments, a chiral-controlled oligonucleotide composition contains non-random or controlled levels of individual oligonucleotide or nucleic acid types. For example, in some embodiments, a chiral-controlled oligonucleotide composition contains one or more oligonucleotide types. In some embodiments, a chiral-controlled oligonucleotide composition contains two or more oligonucleotide types. In some embodiments, a chiral-controlled oligonucleotide composition contains a large number of oligonucleotides. This includes nucleotide types. In some embodiments, a chiral-controlled oligonucleotide composition is a composition of oligonucleotides of a certain oligonucleotide type, the composition comprising multiple oligonucleotides of that oligonucleotide type in non-random or controlled levels.

[0033] Internucleotide bond: As used herein, the term “internucleotide bond” generally refers to a bond that connects nucleoside units of an oligonucleotide or nucleic acid. In some embodiments, the internucleotide bond is a phosphate diester bond (natural phosphate bond (-OP(=O)(OH)O-)), as is widely found in naturally occurring DNA and RNA molecules, which may exist in salt form as will be understood by those skilled in the art. In some embodiments, the internucleotide bond is a modified internucleotide bond (not a natural phosphate bond). In some embodiments, the internucleotide bond is a “modified internucleotide bond,” where at least one oxygen atom or -OH of the phosphate diester bond is replaced by a different organic or inorganic moiety. In some embodiments, such organic or inorganic moiety is selected from =S, =Se, =NR', -SR', -SeR', -N(R')2, B(R')3, -S-, -Se-, and -N(R')-, where each R' is independently as defined and described herein. In some embodiments, the internucleotide bond is a phosphate triester bond, a phosphorothioate bond (or a phosphorothioate diester bond, -OP(=O)(SH)O-, which may exist in salt form as understood by those skilled in the art), or a phosphorothioate triester bond. In some embodiments, the modified internucleotide bond is a phosphorothioate bond. In some embodiments, the internucleotide bond is, for example, a PNA (peptide nucleic acid) or a PMO (phosphodiamidate morpholino oligomer) bond. In some embodiments, the modified internucleotide bond is a non-negatively charged internucleotide bond. In some embodiments, the modified internucleotide bond is a neutral internucleotide bond (e.g., n001 in certain provided oligonucleotides). As those skilled in the art will understand, the internucleotide bond may exist as an anion or a cation at a given pH due to the presence of an acid or base moiety in the bond.In some embodiments, the modified internucleotide linkage is a modified internucleotide linkage designated as s, s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14, s15, s16, s17, and s18 as described in International Publication No. 2017 / 210647.

[0034] In vitro: As used herein, the term “in vitro” refers to an event that occurs not within the body of an organism (e.g., an animal, plant, and / or microorganism), but rather in an artificial environment, such as in a test tube or reactor, under cell culture conditions, etc.

[0035] In vivo: As used herein, the term “in vivo” refers to an event occurring within the body of a living organism (e.g., an animal, a plant, and / or a microorganism).

[0036] Bound phosphorus: As defined herein, the phrase “bound phosphorus” is used to indicate that the particular phosphorus atom mentioned is a phosphorus atom present in an internucleotide bond, which corresponds to the phosphorus atom in a phosphodiester internucleotide bond as present in naturally occurring DNA and RNA. In some embodiments, the bound phosphorus atom is in a modified internucleotide bond, where each oxygen atom of the phosphodiester bond is optionally and independently replaced by an organic or inorganic moiety. In some embodiments, the bound phosphorus atom is P of formula I as described herein. In some embodiments, the bound phosphorus atom is chiral. In some embodiments, the bound phosphorus atom is achiral (e.g., as present in a natural phosphate bond).

[0037] Linker: The term "linker," "binding portion," etc., refers to any chemical portion that links one chemical portion to another. As those skilled in the art will understand, a linker can be divalent, trivalent, or more, depending on the number of chemical portions it links. In some embodiments, the linker is the portion in a polymer that links one oligonucleotide to another. In some embodiments, the linker is optionally located between a terminal nucleoside and a solid support, or between a terminal nucleoside and another nucleoside, nucleotide, or nucleic acid. In some embodiments, in an oligonucleotide, the linker links a chemical portion (e.g., a targeting portion, a lipid portion, a carbohydrate portion, etc.) to an oligonucleotide chain (e.g., at its 5' end, 3' end, a nucleic acid base, a sugar, an internucleotide bond, etc.).

[0038] Modified nucleic acid bases: The terms "modified nucleic acid base" and "modified base" refer to a chemical portion that is chemically different from a nucleic acid base but has the ability to perform at least one function of a nucleic acid base. In some embodiments, a modified nucleic acid base is a nucleic acid base that includes modifications. In some embodiments, a modified nucleic acid base has the ability to perform at least one function of a nucleic acid base, for example, the ability to form a portion in a polymer that has base-pairing ability with nucleic acids containing at least complementary base sequences. In some embodiments, a modified nucleic acid base is a substituted A, T, C, G, or U or a substituted tautomer of A, T, C, G, or U. In some embodiments, a modified nucleic acid base in the context of oligonucleotides refers to a nucleic acid base that is not A, T, C, G, or U.

[0039] Modified Nucleoside: The term "modified nucleoside" refers to a portion of a nucleoside that is derived from or chemically similar to a natural nucleoside but contains chemical modifications that distinguish it from the natural nucleoside. Non-limiting examples of modified nucleosides include those with modifications to bases and / or sugars. Non-limiting examples of modified nucleosides include those with 2' modifications to sugars. Non-limiting examples of modified nucleosides include debasic nucleosides (which lack nucleic acid bases). In some embodiments, a modified nucleoside has the ability to form a portion in a polymer having the ability to perform at least one function of a nucleoside, for example, the ability to base pair with nucleic acids containing at least a complementary base sequence.

[0040] Modified nucleotides: The term "modified nucleotide" includes any chemical portion that is structurally different from a natural nucleotide but has the ability to perform at least one function of a natural nucleotide. In some embodiments, modified nucleotides include modifications to sugars, bases, and / or internucleotide bonds. In some embodiments, modified nucleotides include modified sugars, modified nucleic acid bases, and / or modified internucleotide bonds. In some embodiments, modified nucleotides have the ability to perform at least one function of a nucleotide, for example, the ability to form subunits in a polymer having the ability to base pair with nucleic acids containing at least complementary base sequences.

[0041] Modified sugar: The term “modified sugar” refers to a portion of a sugar that can be replaced. Modified sugars mimic the spatial arrangement, electronic properties, or any other physicochemical properties of a sugar. In some embodiments, as described herein, the modified sugar is a substituted ribose or deoxyribose. In some embodiments, the modified sugar includes 2'-modifications. Useful examples of 2'-modifications are widely available in the art and are described herein. In some embodiments, the 2'-modification is 2'-OR, where R is optionally substituted C. 1~10It is aliphatic. In some embodiments, the 2'-modification is 2'-OMe. In some embodiments, the 2'-modification is 2'-MOE. In some embodiments, the modified sugar is a bicyclic sugar (e.g., a sugar used in LNA, BNA, etc.). In some embodiments, in the context of oligonucleotides, the modified sugar is a sugar other than ribose or deoxyribose, as is typically found in natural RNA or DNA.

[0042] Nucleic acids: As used herein, the term “nucleic acid” includes any nucleotide and its polymers. The term “polynucleotide” as used herein refers to nucleotides in polymeric form of any length, whether ribonucleotides (RNA) or deoxyribonucleotides (DNA) or any combination thereof. These terms refer to the primary structure of molecules and thus include double-stranded and single-stranded DNA and double-stranded and single-stranded RNA. These terms include, but are not limited to, analogues of RNA or DNA containing modified nucleotides and / or modified polynucleotides, such as methylated, protected, and / or capped nucleotides or polynucleotides. These terms encompass poly- or oligo-ribonucleotides (RNA) and poly- or oligo-deoxyribonucleotides (DNA); RNA or DNA derived from N-glycosides or C-glycosides of nucleic acid bases and / or modified nucleic acid bases; nucleic acids derived from sugars and / or modified sugars; and nucleic acids derived from phosphate crosslinks and / or modified internucleotide links. This term encompasses nucleic acids containing any combination of nucleic acid bases, modified nucleic acid bases, sugars, modified sugars, phosphate crosslinks, or modified internucleotide bonds. Examples include, but are not limited to, nucleic acids containing a ribose moiety, nucleic acids containing a deoxyribose moiety, nucleic acids containing both a ribose moiety and a deoxyribose moiety, and nucleic acids containing a ribose moiety and a modified ribose moiety. Unless otherwise specified, the prefix "poly-" refers to nucleic acids containing 2 to approximately 10,000 nucleotide monomer units, and the prefix "oligo-" refers to nucleic acids containing 2 to approximately 200 nucleotide monomer units.

[0043] Nucleic acid bases: The term "nucleic acid base" refers to the parts of nucleic acids involved in hydrogen bonding, which sequence-specifically connects one nucleic acid chain to another complementary chain. The most common naturally occurring nucleic acid bases are adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, naturally occurring nucleic acid bases are modified adenine, guanine, uracil, cytosine, or thymine. In some embodiments, naturally occurring nucleic acid bases are methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, the nucleic acid bases include a heteroaryl ring in which the ring atom is nitrogen and, when in a nucleoside, that nitrogen is bonded to a sugar moiety. In some embodiments, the nucleic acid bases include a heterocyclic ring in which the ring atom is nitrogen and, when in a nucleoside, that nitrogen is bonded to a sugar moiety. In some embodiments, the nucleic acid base is a “modified nucleic acid base,” i.e., a nucleic acid base other than adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, the modified nucleic acid base is a substituted A, T, C, G, or U. In some embodiments, the modified nucleic acid base is a substituted tautomer of A, T, C, G, or U. In some embodiments, the modified nucleic acid base is a methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, the modified nucleic acid base mimics the spatial arrangement, electronic properties, or any other physicochemical properties of the nucleic acid base and retains the properties of hydrogen bonding, which sequence-specifically binds one nucleic acid chain to another. In some embodiments, the modified nucleic acid base can pair with all five naturally occurring bases (uracil, thymine, adenine, cytosine, or guanine) without substantially affecting melting behavior, recognition by intracellular enzymes, or the activity of the oligonucleotide double helix. As used herein, the term “nucleic acid base” also includes structural analogues used in place of natural or naturally occurring nucleotides, such as modified nucleic acid bases and nucleic acid base analogues. In some embodiments, the nucleic acid base is A, T, C, G, or U, or a tautomer that is optionally substituted with A, T, C, G, or U.In some embodiments, “nucleic acid base” refers to a nucleic acid base unit in an oligonucleotide or nucleic acid (for example, A, T, C, G, or U, as found in an oligonucleotide or nucleic acid).

[0044] Nucleoside: The term "nucleoside" refers to a nucleic acid base or modified nucleic acid base that is combined with a sugar or modified sugar. This refers to a covalently bonded portion. In some embodiments, the nucleoside is a natural nucleoside, e.g., adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, or deoxycytidine. In some embodiments, the nucleoside is a modified nucleoside, e.g., a substituted natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine. In some embodiments, the nucleoside is a substituted tautomer of a modified nucleoside, e.g., a natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine. In some embodiments, "nucleoside" refers to a nucleoside unit in an oligonucleotide or nucleic acid.

[0045] Nucleotide: As used herein, the term “nucleotide” refers to the monomeric unit of a polynucleotide, which consists of a nucleic acid base, a sugar, and one or more internucleotide links (e.g., a phosphate linkage in natural DNA and RNA). Naturally occurring bases [guanine, (G), adenine, (A), cytosine, (C), thymine, (T), and uracil (U)] are derivatives of purines or pyrimidines, but it should be understood that this also includes naturally occurring and non-naturally occurring base analogs. Naturally occurring sugars are pentose (five-carbon sugar) deoxyribose (which forms DNA) or ribose (which forms RNA), but it should be understood that this also includes naturally occurring and non-naturally occurring sugar analogs. Nucleotides are joined by internucleotide links to form nucleic acids or polynucleotides. Numerous internucleotide links are known in the art (but are not limited to phosphates, phosphorothioates, boranophosphates, etc.). Artificial nucleic acids include PNA (peptide nucleic acid), phosphate triesters, phosphorothionic acid, H-phosphonic acid, phosphoramidic acid, boranophosphate, methylphosphonic acid, phosphonoacetic acid, thiophosphonoacetic acid, and other variants of the phosphate skeleton of natural nucleic acids, as described herein. In some embodiments, natural nucleotides include naturally occurring bases, sugars, and internucleotide bonds. As used herein, the term “nucleotide” also includes structural analogues used in place of natural or naturally occurring nucleotides, such as modified nucleotides and nucleotide analogues. In some embodiments, “nucleotide” refers to an oligonucleotide or a nucleotide unit in a nucleic acid.

[0046] Oligonucleotides: The term "oligonucleotide" refers to polymers or oligomers of nucleotides, and may encompass any combination of natural and unnatural nucleic acid bases, sugars, and internucleotide bonds.

[0047] Oligonucleotides can be single-stranded or double-stranded. Single-stranded oligonucleotides may have a double-stranded region (formed by two parts of a single-stranded oligonucleotide), and double-stranded oligonucleotides containing two oligonucleotide strands may have a single-stranded region, for example, in a region where the two oligonucleotide strands are not complementary to each other. Exemplary oligonucleotides include, but are not limited to, structural genes, genes containing regulatory and termination regions, self-replicating systems such as viral DNA or plasmid DNA, single-stranded and double-stranded RNAi agents and other RNA interference reagents (RNAi agents or iRNA agents), shRNA, antisense oligonucleotides, ribozymes, microRNAs, microRNA mimetic compounds, supermirs, aptamers, antimirs, antagonistmirs, Ul adapters, triple-stranded oligonucleotides, G quadruple-stranded oligonucleotides, RNA activators, immunostimulatory oligonucleotides, and decoy oligonucleotides.

[0048] The oligonucleotides of this disclosure may vary in length. In detailed embodiments, the oligonucleotides may range in length from about 2 to about 200 nucleosides. Various related embodiments In some embodiments, the single-stranded, double-stranded, or triple-stranded oligonucleotide can range in length from about 4 to about 10 nucleosides, about 10 to about 50 nucleosides, about 20 to about 50 nucleosides, about 15 to about 30 nucleosides, about 20 to about 30 nucleosides. In some embodiments, the oligonucleotide is about 9 to about 39 nucleosides in length. In some embodiments, the oligonucleotide is at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nucleosides in length. In some embodiments, the oligonucleotide is 19 nucleosides in length. In some embodiments, the oligonucleotide is at least 20 nucleosides in length. In some embodiments, the oligonucleotide is at least 25 nucleosides in length. In some embodiments, the oligonucleotide is at least 30 nucleosides in length. In some embodiments, the oligonucleotide is a double-strand of a complementary strand that is at least 18 nucleosides in length. In some embodiments, the oligonucleotide is a double-strand of a complementary strand that is at least 21 nucleosides in length. In some embodiments, each nucleoside counted towards the oligonucleotide length independently comprises A, T, C, G or U, or A, T, C, G or U optionally substituted, or tautomers optionally substituted of A, T, C, G or U.

[0049] Oligonucleotide type: As used herein, the phrase "oligonucleotide type" is used to define an oligonucleotide having a particular base sequence, pattern of backbone linkages (i.e., internucleotide linkage type, e.g., pattern of phosphate, phosphorothioate, phosphorothioate triester, etc.), pattern of backbone chiral centers [i.e., pattern of linked phosphorus stereochemistry (Rp / Sp)], and pattern of backbone phosphorus modifications (e.g., pattern of "-XLR 1 " groups in Formula I as described herein). In some embodiments, oligonucleotides of a common designated "type" are structurally identical to each other.

[0050] Those skilled in the art will understand that the synthesis methods of the present disclosure provide a degree of control in oligonucleotide chain synthesis such that each nucleotide unit of the oligonucleotide chain can be pre-designed and / or selected to have a specific stereochemistry and / or specific modification of the bound phosphorus, and / or a specific base and / or a specific sugar. In some embodiments, the oligonucleotide chain is pre-designed and / or selected to have a specific combination of stereocenters in the bound phosphorus. In some embodiments, the oligonucleotide chain is designed and / or determined to have a specific combination of modifications in the bound phosphorus. In some embodiments, the oligonucleotide chain is designed and / or selected to have a specific combination of bases. In some embodiments, the oligonucleotide chain is designed and / or selected to have one or more specific combinations of the above structural properties. In some embodiments, the present disclosure provides a composition comprising or consisting of a plurality of oligonucleotide molecules (e.g., a chiral-controlled oligonucleotide composition). In some embodiments, all such molecules are of the same type (i.e., structurally identical to one another). However, in some embodiments, the composition provided comprises a plurality of oligonucleotides of different types, typically in predetermined relative amounts.

[0051] Optionally Substituted: As described herein, the compounds of this disclosure, e.g., oligonucleotides, may include optionally substituted and / or substituted moieties. Generally, the term “substituted,” whether preceded by the term “optionally,” means that one or more hydrogens of the indicating moiety are replaced with preferred substituents. Unless otherwise specified, an “optionally substituted” group may have preferred substituents at each of its substituted positions, and when two or more positions in any given structure can be replaced with two or more substituents selected from a specified group, the substituents may be the same or different at each position. In some embodiments, an optionally substituted group is substituted Not provided. The substituent combinations envisioned in this disclosure preferably result in the formation of stable or chemically feasible compounds. The term “stable,” as used herein, means a compound that remains substantially unchanged when subjected to conditions that enable its generation, detection, and, in certain embodiments, its recovery, purification, and use for one or more purposes disclosed herein. Specific substituents are described below.

[0052] Substitutable atoms, for example, preferred monovalent substituents on preferred carbon atoms, are independently halogens;-(CH2) 0~4 R 〇 ;-(CH2) 0~4 Ure 〇 ;-O(CH2) 0~4 R o -O-(CH2) 0~4 C(O)OR 〇 ;-(CH2) 0~4 CH(OR 〇 )2;R 〇 -(CH2) can be substituted with 0~4 Ph;R 〇 -(CH2) can be substituted with 0~4 O(CH2) 0~1 Ph;R 〇 -CH=CHPh;R can be substituted with 〇 -(CH2) can be substituted with 0~4 O(CH2) 0~1 -Pyridyl;-NO2;-CN;-N3;-(CH2) 0~4 N(R 〇 )2;-(CH2) 0~4 N(R 〇 )C(O)R 〇 ;-N(R 〇 )C(S)R 〇 ;-(CH2) 0~4 N(R 〇 )C(O)NR 〇 2;-N(R 〇 )C(S)NR 〇 2;-(CH2) 0~4 N(R 〇 )C(O)OR 〇 ;-N(R 〇 )N(R 〇 )C(O)R〇 ;-N(R 〇 )N(R 〇 )C(O)NR 〇 2;-N(R 〇 )N(R 〇 )C(O)OR 〇 ;-(CH2) 0~4 C(O)R 〇 ;-C(S)R 〇 ;-(CH2) 0~4 C(O)OR 〇 ;-(CH2) 0~4 C(O)SR 〇 ;-(CH2) 0~4 C(O)OSiR 〇 3;-(CH2) 0~4 OC(O)R 〇 ;-OC(O)(CH2) 0~4 SR 〇 、-SC(S)SR 〇 ;-(CH2) 0~4 SC(O)R 〇 ;-(CH2) 0~4 C(O)NR 〇 2;-C(S)NR 〇 2;-C(S)SR 〇 ;-(CH2) 0~4 OC(O)NR 〇 2;-C(O)N(OR 〇 )R 〇 ;-C(O)C(O)R 〇 ;-C(O)CH2C(O)R 〇 ;-C(NOR 〇 )R 〇 ;-(CH2) 0~4 SSR 〇 ;-(CH2) 0~4 S(O)2R 〇 ;-(CH2) 0~4 S(O)2OR 〇 ;-(CH2) 0~4 OS(O)2R 〇 ;-S(O)2NR 〇 2;-(CH2) 0~4 S(O)R 〇 ;-N(R 〇 )S(O)2NR 〇 2;-N(R 〇 )S(O)2R 〇 ;-N(OR 〇)R 〇 ;-C(NH)NR 〇 2;-Si(R 〇 )3;-OSi(R 〇 )3;-B(R 〇 )2;-OB(R 〇 )2;-OB(OR 〇 )2;-P(R 〇 )2;-P(OR 〇 )2;-P(R 〇 )(OR 〇 );-OP(R 〇 )2;-OP(OR 〇 )2;-OP(R 〇 )(OR 〇 );-P(O)(R 〇 )2;-P(O)(OR 〇 )2;-OP(O)(R 〇 )2;-OP(O)(OR 〇 )2;-OP(O)(OR 〇 )(SR 〇 );-SP(O)(R 〇 )2;-SP(O)(OR 〇 )2;-N(R 〇 )P(O)(R 〇 )2;-N(R 〇 )P(O)(OR 〇 )2;-P(R 〇 )2[B(R 〇 )3];-P(OR 〇 )2[B(R 〇 )3];-OP(R 〇 )2[B(R 〇 )3];-OP(OR 〇 )2[B(R 〇 )3];-(C 1~4 linear or branched alkylene)O-N(R 〇 )2; or -(C 1~4 linear or branched alkylene)C(O)O-N(R 〇 )2, wherein each R 〇 is optionally substituted as defined herein and is independently hydrogen, C<00(00137>aliphatic, C 1~20 heteroaliphatic having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon and phosphorus, -CH2-(C 6~14 aryl), -O(CH2)0~1 (C 6~14 aryl), -CH2- (5-14 member heteroaryl ring), a 5-20 member monocyclic, bicyclic, or polycyclic saturated, partially unsaturated or aryl ring having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus, or notwithstanding the above definition, R 〇 Two independent entities, together with one or more intervening atoms, form a 5-20 member monocyclic, bicyclic, or polycyclic saturated, partially unsaturated, or aryl ring having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus, which may be substituted as defined below.

[0053] R 〇 (or R 〇 Suitable monovalent substituents on the ring formed by the two independent entities of -(CH2) together with their intervening atoms are, independently, halogens, -(CH2) 0~2 R ● ,-(HaroR ● ), -(CH2) 0~2 OH, -(CH2) 0~2 Ure ● ,-(CH2) 0~2 CH(OR ● )2;-O(HaroR ● ), -CN, -N3, -(CH2) 0~2 C(O) R ● ,-(CH2) 0~2 C(O)OH, -(CH2) 0~2 C(O)OR ● ,-(CH2) 0~2 SR ● ,-(CH2) 0~2 SH, -(CH2) 0~2 NH2, -(CH2) 0~2 NHR ● ,-(CH2) 0~2 NR ● 2, -NO2, -SiR ● 3. -OSiR ● 3, -C(O)SR ● ,-(C 1~4 Linear or branched alkylene)C(O)OR ● , or -SSR ●And in the formula, each R ● It is either unsubstituted, or if preceded by "halo", it is substituted by only one or more halogens, independently, C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 The pH is independently selected from a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms selected from nitrogen, oxygen, and sulfur. 〇 Suitable divalent substituents on the saturated carbon atom include =O and =S.

[0054] For example, suitable divalent substituents on suitable carbon atoms are independently: =O, =S, =NNR * 2. =NNHC(O)R * ,=NNHC(O)OR * ,=NNHS(O)2R * ,=NR * 、=NOR * , -O(C(R * 2)) 2~3 O-, or -S(C(R * 2)) 2~3 S-, and in the formula, R * Each independent entity is a hydrogen atom, which can be substituted as defined below. 1~6 Selected from aliphatic and unsubstituted 5-6 member saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Suitable divalent substituents bonded to the vicinal substituted carbon of the "optionally substituted" group include -O(CR * 2) 2~3 O- is mentioned, and in the formula, R * Each independent entity is a hydrogen atom, which can be substituted as defined below. 1~6 The compounds are selected from aliphatic and unsubstituted 5-6 member saturated, partially unsaturated, and aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0055] R * Preferred substituents on the aliphatic group are, independently, halogens, -R ● ,-(HaroR ● ), -OH, -OR ● ,-O(HaroR● ), -CN, -C(O)OH, -C(O)OR ● -NH2, -NHR ● , -NR ● 2, or -NO2, where each R ● It is either unsubstituted, or if preceded by "halo", it is substituted by only one or more halogens, independently, C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 The pH is a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0056] In some embodiments, suitable substituents on the replaceable nitrogen are independently -R † , -NR † 2, -C(O)R † , -C(O)OR † ,-C(O)C(O)R † -C(O)CH2C(O)R † -S(O)2R † -S(O)2NR † 2, -C(S)NR † 2, -C(NH)NR † 2, or -N(R † )S(O)2R † And; in the formula, each R † These are independently hydrogen, and C which can be substituted as defined below. 1~6 Aliphatic, unsubstituted-OPh, or an unsubstituted 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or notwithstanding the above definition, R † Two independent entities, together with one or more intervening atoms, form an unsubstituted 3-12 member saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0057] R † Preferred substituents on the aliphatic group are, independently, halogens, -R ● ,-(HaroR ● ), -OH, -OR ●,-O(HaroR ● ), -CN, -C(O)OH, -C(O)OR ● -NH2, -NHR ● , -NR ● 2, or -NO2, where each R ● It is either unsubstituted, or if preceded by "halo", it is substituted by only one or more halogens, independently, C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 The pH is a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0058] Oral: When the terms "oral administration" and "administered orally" are used herein, In the sense understood within this technical field, it refers to the oral administration of a compound or composition.

[0059] Parenteral: When used herein, the terms “parenteral administration” and “administered parenterally” mean, as understood in the art, a mode of administration other than enteral and topical administration, usually by injection, and without limitation include intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injections and infusions.

[0060] Partially unsaturated: As used herein, the term “partially unsaturated” refers to a ring moiety containing at least one double or triple bond. The term “partially unsaturated” is intended to encompass rings having multiple unsaturated moies, but not to include aryl or heteroaryl moies as defined herein.

[0061] Pharmaceutical Composition: As used herein, the term “pharmaceutical composition” refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose quantity appropriate for administration in a therapeutic regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition may be specifically formulated for administration in solid or liquid form, including: oral administration, e.g., oral tablets (aqueous or non-aqueous solutions or suspensions), tablets, e.g., buccal, sublingual, and systemically absorbed, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., sterile solutions or suspensions, or as sustained-release formulations, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection; topical application, e.g., as creams, ointments, or controlled-release patches or sprays applied to the skin, lungs, or oral cavity; e.g., as pessaries, creams, or foams, for vaginal or rectal, sublingual, intraocular, transdermal, or adapted for the nasal cavity, lungs, and other mucosal surfaces.

[0062] Pharmacopoeia-acceptable: As used herein, the term "pharmacopoeia-acceptable" means, within reasonable medical judgment, a compound, material, composition and / or dosage form suitable for use in contact with human and animal tissues, without excessive toxicity, irritation, allergic reactions, or other problems or complications, as is appropriate for a reasonable risk-benefit ratio.

[0063] Pharmacopoeia-acceptable carrier: As used herein, the term “pharmacopoeia-acceptable carrier” means a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, or solvent, that encapsulates a material and is involved in the transport or delivery of a compound of interest from one organ or part of the body to another organ or part of the body. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the formulation and is not harmful to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; tragacanth powder; malt; gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic and suitable substances used in pharmaceutical formulations.

[0064] Pharmacopoeia-acceptable salt: The term "pharmacopoeia-acceptable salt" is used herein when This refers to salts of such compounds that are appropriate for use in a pharmaceutical context, i.e., salts that, within reasonable medical judgment, do not cause excessive toxicity, irritation, or allergic reactions, are suitable for use in contact with human and lower animal tissues, and have a reasonable risk-benefit ratio. Pharmaceutically acceptable salts are well known in the art. For example, S.M. Berge, et al. For details on pharmaceutically acceptable salts, see J. Pharmaceutical Sciences, 66: 1-19 (1977). This is described in detail. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts, which are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by other methods used in the art, such as ion exchange. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, adipine salts, alginates, ascorbic acid, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphor sulfons, citrates, cyclopentanepropionates, digluconates, dodecyl sulfates, ethanesulfonates, formates, fumarates, glucoheptonates, glycerophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxy- Examples include ethanesulfonates, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malons, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propions, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, and valersates. In some embodiments, the compounds provided include one or more acidic groups, such as oligonucleotides, and pharmaceutically acceptable salts are alkali salts, alkaline earth metal salts, or ammonium salts (e.g., ammonium salts of N(R)3 [wherein each R is independently defined and described herein]). Typical alkali salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium. In some embodiments, the pharmaceutically acceptable salt is a sodium salt. In some embodiments, the pharmaceutically acceptable salt is a potassium salt. In some embodiments, the pharmaceutically acceptable salt is a calcium salt.In some embodiments, pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls, sulfons, and arylsulfons having 1 to 6 carbon atoms. In some embodiments, the provided compounds comprise two or more acidic groups; for example, oligonucleotides may comprise two or more acidic groups (e.g., in a natural phosphate bond and / or a modified internucleotide bond). In some embodiments, a pharmaceutically acceptable salt of such a compound, or a salt in general, comprises two or more cations, which may be the same or different. In some embodiments, in a pharmaceutically acceptable salt (or a salt in general), all ionizable hydrogens in the acidic groups (e.g., in aqueous solutions with pKas of about 11, 10, 9, 8, 7, 6, 5, 4, 3 or 2 or less; in some embodiments, about 7 or less; in some embodiments, about 6 or less; in some embodiments, about 5 or less; in some embodiments, about 4 or less; in some embodiments, about 3 or less) are replaced by cations. In some embodiments, each phosphorothioate and phosphate group exists independently in its salt form (e.g., in the case of the sodium salt, -OP(O)(SNa)-O- and -OP(O)(ONa)-O-, respectively). In some embodiments, each phosphorothioate and phosphate internucleotide bond exists independently in its salt form (e.g., in the case of the sodium salt, -OP(O)(SNa)-O- and -OP(O)(ONa)-O-, respectively). In some embodiments, the pharmaceutically acceptable salt is the sodium salt of the oligonucleotide. In some embodiments, the pharmaceutically acceptable salt is the sodium salt of the oligonucleotide, where each acidic phosphate group and modified phosphate group (e.g., phosphorothioate, phosphate, etc.) exists in its respective form. It exists as a compound, or as a salt (all sodium salts).

[0065] Protecting group: When the term "protecting group" is used herein, it refers to a term well known in the art, as seen in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd For more details, see edition, John Wiley & Sons, 1999 (this entire edition is incorporated herein by reference). The following are listed in detail. Also, Current Protocols in Nucleic Acid Chemistry, edited by Serge L. Beaucage et al. 06 / 2012 (the entirety of Chapter 2 is referenced herein). Other examples include protecting groups specifically adapted to nucleoside and nucleotide chemistry as described herein (referred to herein by reference). Preferred amino protecting groups include, but are not limited to, those described herein and / or International Publication Nos. 2018 / 022473, 2018 / 098264, 2018 / 223056, 2018 / 223073, 2018 / 223081, 2018 / 237194, 2019 / 032607, 2019 / 055951, and / or International Publication Nos. 2019 / 075357, or U.S. Provisional Patent Application Nos. 62 / 825766 and U.S. Provisional Patent Application Nos. 62 / 911339 (the descriptions of each of these protecting groups are referred to herein by reference independently).

[0066] Sample: As used herein, the term “sample” typically refers to an aliquot of material obtained from or derived from the source of interest. In some embodiments, the source of interest is a biological or environmental source. In some embodiments, the source of interest may be or include cells or organisms, such as microorganisms, plants, or animals (e.g., humans). In some embodiments, the source of interest may be or include biological tissue or biological fluids. In some embodiments, biological tissue or bodily fluid may be or include amniotic fluid, aqueous humor, ascites, bile, bone marrow, blood, breast milk, cerebrospinal fluid, earwax, chyle, porridge, ejaculate, endolymph, exudate, feces, gastric acid, gastric juice, lymph, mucus, pericardial fluid, perilymph, ascites, pleural fluid, pus, catarrhal secretions, saliva, sebum, semen, serum, smegma, sputum, synovial fluid, sweat, tears, urine, vaginal secretions, vitreous fluid, vomit, and / or combinations thereof or one or more components thereof. In some embodiments, biological fluid may be or include intracellular fluid, extracellular fluid, intravascular fluid (plasma), interstitial fluid, lymph, and / or cell permeable fluid. In some embodiments, biological fluid may be or include plant exudate. In some embodiments, biological tissue or biological specimens are subjected to, for example, aspiration, biopsy (e.g., fine needle or tissue biopsy), swab (e.g., oral cavity, nasal cavity, skin, or vaginal swab), scraping, surgery, washing, or lavage (e.g., bronchoalveoli, tubules, nasal cavity, eyeballs, oral cavity, uterus, etc.) It can be obtained through vaginal or other washing or lavage. Some practices Morphologically, a biological sample is or contains cells obtained from an individual. In some embodiments, the sample is a “primary sample” obtained directly from the source of interest by any suitable means. In some embodiments, as will be apparent from the context, the term “sample” refers to a preparation obtained by processing the primary sample (e.g., by removing one or more of its components and / or by adding one or more agents to it), e.g., filtration using a semipermeable membrane. Such a “processed sample” may contain nucleic acids or proteins obtained, for example, extracted from the sample or by subjecting the primary sample to one or more techniques such as nucleic acid amplification or reverse transcription, isolation and / or purification of specific components.

[0067] Subject: As used herein, the term “subject” or “test subject” means in this disclosure any organism to which the provided compound (e.g., the provided oligonucleotide) or composition is administered, for example, for experimental, diagnostic, preventive and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans; insects; worms, etc.) and plants. In some embodiments, the subject is human. In some embodiments, the subject may have and / or be susceptible to a disease, disorder and / or pathology.

[0068] Substantially: As used herein, the term “substantially” refers to a qualitative condition that exhibits the entire or nearly entire range or degree of the desired feature or characteristic. A nucleotide sequence substantially complementary to a second sequence is not identical to the second sequence, but is approximately or nearly identical. In addition, those skilled in the art of biology and / or chemistry will understand that it is rare, if any, for biological and chemical phenomena to go to completion and / or progress to completeness or to achieve or avoid absolute results. Thus, the term “substantially” is used herein to capture the potential lack of completeness inherent in many biological and / or chemical phenomena.

[0069] Sugars: The term "sugar" refers to closed-chain and / or open-chain monosaccharides or polysaccharides. In some embodiments, the sugar is a monosaccharide. In some embodiments, the sugar is a polysaccharide. Examples of sugars, but not limited to, include ribose, deoxyribose, pentofuranose, pentopyranose, and hexopyranose moieties. As used herein, the term "sugar" also includes structural analogs used in place of conventional sugar molecules such as glycols, whose polymers form the backbone of nucleic acid analogs, glycol nucleic acids ("GNAs"), etc. As used herein, the term "sugar" also includes structural analogs used in place of natural or naturally occurring nucleotides, such as modified sugars and nucleotide sugars. In some embodiments, the sugar is an RNA or DNA sugar (ribose or deoxyribose). In some embodiments, the sugar is a modified ribose or deoxyribose sugar, e.g., 2'-modified, 5'-modified, etc. When used in oligonucleotides and / or nucleic acids as described herein, in some embodiments, modified sugars may provide one or more desired properties, activities, etc. In some embodiments, the sugar is optionally substituted ribose or deoxyribose. In some embodiments, “sugar” refers to a sugar unit in an oligonucleotide or nucleic acid.

[0070] Susceptible to: Individuals "susceptible to" a disease, disorder, and / or condition are individuals with a higher risk of developing the disease, disorder, and / or condition compared to members of the general public. In some embodiments, individuals susceptible to a disease, disorder, and / or condition have a predisposition to that disease, disorder, and / or condition. In some embodiments, individuals susceptible to a disease, disorder, and / or condition may not have been diagnosed with that disease, disorder, and / or condition. In some embodiments, individuals susceptible to a disease, disorder, and / or condition may exhibit symptoms of that disease, disorder, and / or condition. In some embodiments, individuals susceptible to a disease, disorder, and / or condition may not exhibit symptoms of that disease, disorder, and / or condition. In some embodiments, individuals susceptible to a disease, disorder, and / or condition will develop that disease, disorder, and / or condition. In some embodiments, individuals susceptible to disease, disorder, and / or pathology will not develop that disease, disorder, and / or pathology.

[0071] Therapeutic Agent: As used herein, the term “therapeutic agent” generally refers to any agent that, when administered to a subject, produces a desired effect (e.g., a desired biological, clinical, or pharmacological effect). In some embodiments, an agent is considered a therapeutic agent if it demonstrates a statistically significant effect in an appropriate population. In some embodiments, an appropriate population is a population of subjects suffering from and / or susceptible to a disease, disorder, or condition. In some embodiments, an appropriate population is a population of model organisms. In some embodiments, an appropriate population may be defined by one or more criteria, such as age group, sex, genetic background, pre-existing clinical condition, or history of exposure to a therapy. In some embodiments, a therapeutic agent is a substance that, when administered to a subject in an effective dose, alleviates, improves, reduces, inhibits, prevents, delays the onset of, reduces the severity of, and / or decreases the incidence of one or more symptoms or characteristics of the disease, disorder, and / or condition in question. In some embodiments, “therapeutic agent” is a drug that has received approval from a government agency or is required to receive such approval before it can be placed on the market for administration to humans. In some embodiments, the “therapeutic agent” is a drug that requires a physician’s prescription for administration to humans. In some embodiments, the therapeutic agent is a compound provided, for example, a provided oligonucleotide.

[0072] Therapeutic dose: As used herein, the term “therapeutic dose” means the amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that, when administered as part of a therapeutic regimen, elicits a desired biological response. In some embodiments, the therapeutic dose of a substance is the amount sufficient to treat, diagnose, prevent, and / or delay the onset of a disease, disorder, and / or condition when administered to a subject suffering from or susceptible to that disease, disorder, and / or condition. As those skilled in the art will understand, the effective dose of a substance may vary depending on factors such as the desired biological endpoint, the substance to be delivered, the target cells or tissues, etc. For example, the effective dose of a compound in a formulation for treating a disease, disorder, and / or condition is the amount that alleviates, improves, reduces, inhibits, prevents, delays the onset, reduces the severity, and / or decreases the incidence of one or more symptoms or characteristics of that disease, disorder, and / or condition. In some embodiments, the therapeutic dose is administered in a single dose; in some embodiments, multiple unit doses are required to deliver the therapeutic dose.

[0073] To treat: As used herein, the terms “to treat,” “treatment,” or “to treat” mean any method used to partially or completely alleviate, improve, reduce, inhibit, prevent, delay the onset, reduce the severity, and / or decrease the incidence of one or more symptoms or features of a disease, disorder, and / or condition. Treatment may be administered to subjects who are not exhibiting signs of a disease, disorder, and / or condition. In some embodiments, treatment may be administered to subjects exhibiting only the initial signs of a disease, disorder, and / or condition, for example, to reduce the risk of developing lesions associated with that disease, disorder, and / or condition.

[0074] Unsaturated: The term "unsaturated," as used herein, means that a part has one or more unsaturated units.

[0075] Wild-type: As used herein, the term “wild-type” has the meaning understood in the art to refer to an entity having the structure and / or activity as found in its natural and “normal” state or context (as opposed to mutant, diseased, modified, etc.). Those skilled in the art will understand that wild-type genes and polypeptides often exist in multiple different forms (e.g., alleles).

[0076] As those skilled in the art will understand, the methods and compositions described herein with respect to the provided compounds (e.g., oligonucleotides) are generally also applicable to pharmaceutically acceptable salts of such compounds.

[0077] Description of a specific embodiment Oligonucleotides provide a useful tool for a wide range of applications. For example, MAPT oligonucleotides are useful in therapeutic, diagnostic, and research applications, including, but not limited to, the treatment of various MAPT-related conditions, disorders, and diseases, including Alzheimer's disease (AD) and frontotemporal dementia (FTD). The use of naturally occurring nucleic acids (e.g., unmodified DNA or RNA) is limited by their vulnerability to, for example, endonucleases and exonucleases. Therefore, various synthetic counterparts have been developed to circumvent these drawbacks and / or improve various properties and activities. This includes synthetic oligonucleotides that involve chemical modifications, such as base modifications, sugar modifications, and skeletal modifications, to make such molecules less susceptible to degradation and to improve other properties and / or activities. From this perspective, modifications to internucleotide bonds can introduce chirality, and certain properties can be influenced by the stereochemistry of the phosphorus atoms bound to the oligonucleotide. For example, binding affinity, sequence-specific binding to complementary RNA, stability to nucleases, cleavage of target nucleic acids, delivery, and pharmacokinetics can be particularly influenced by the chirality of the phosphorus atoms bound to the backbone.

[0078] In some embodiments, the MAPT oligonucleotide contains a sequence that is completely or substantially identical to, or completely or substantially complementary to, 10 or more (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more) adjacent bases of the MAPT genome sequence or a transcript therefrom (e.g., mRNA (e.g., premRNA, post-splicing mRNA, etc.)). In some embodiments, the MAPT oligonucleotide contains a sequence that is completely complementary to 10 or more (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more) adjacent bases of the MAPT transcript. In some embodiments, the number of adjacent bases is approximately 15 to 20. In some embodiments, the number of adjacent bases is approximately 20. In some embodiments, oligonucleotides targeting MAPT can hybridize with MAPT transcripts (e.g., premRNA, RNA, etc.) and reduce the levels of MAPT transcripts and / or proteins encoded by MAPT transcripts.

[0079] In some embodiments, the Disclosure provides MAPT oligonucleotides as disclosed herein, for example, in the Table. In some embodiments, the Disclosure provides MAPT oligonucleotides having a nucleotide sequence as disclosed herein, for example, in the Table, or a portion thereof containing at least 10 (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more) adjacent nucleotides, wherein the MAPT oligonucleotide is sterically random or not chirally controlled, and in the formula, each T may be independently substituted with U, and vice versa.

[0080] In some embodiments, the internucleotide links of the oligonucleotide include or consist of 1-5, 1-10, 1-15, 1-20, 1-25, 1-30, 1-40, 1-50 or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more chiral-controlled internucleotide links. In some embodiments, the disclosure provides a MAPT oligonucleotide composition in which the MAPT oligonucleotide comprises at least one chiral-controlled internucleotide link. In some embodiments, the disclosure provides a MAPT oligonucleotide composition in which the MAPT oligonucleotide is sterically random or not chiral-controlled. In some embodiments, in the MAPT oligonucleotide, at least one internucleotide link is sterically random and at least one internucleotide link is chiral-controlled.

[0081] In some embodiments, the internucleotide bond of the oligonucleotide comprises or consists of one or more negatively charged internucleotide bonds (e.g., phosphorothioate internucleotide bonds, natural phosphate bonds, etc.). In some embodiments, the disclosure relates to MAPT oligonucleotides comprising at least one neutral or non-negatively charged internucleotide bond as described herein.

[0082] MAPT In some embodiments, MAPT refers to a gene or gene product from any species (including, but not limited to, nucleic acids, transcripts, and proteins encoded therein, including DNA or RNA; any form of MAPT, e.g., from a wild-type or mutant allele), MAPT, TAU, MSTD, PPND, D It may be known as DPAC, MAPTL, MTBT1, MTBT2, FTDP-17, or PPP1R103. In some embodiments, it refers to a gene and its product in humans. In some embodiments, it refers to a gene and its product in non-human primates. Various MAPT sequences from humans, mice, rats, monkeys, etc., including their variants, are readily available to those skilled in the art. In some embodiments, MAPT is human or mouse MAPT, which is wild-type or mutant. MAPT has been reported to have several functions. Various techniques, such as assays, cell and animal models, have also been reported and can be used to characterize and / or evaluate the techniques provided in this disclosure (e.g., oligonucleotides, compositions, methods, etc.).

[0083] In some embodiments, the MAPT gene, transcript (e.g., pre- or post-splicing mRNA), or protein variant or isoform may include mutations. In some embodiments, the MAPT gene, transcript, or protein may be a variant or isoform that has undergone alternative splicing, or its transcript or translation product.

[0084] MAPT-related conditions, disorders, or diseases Various pathological conditions, disorders, or diseases have been reported to be associated with MAPT. Generally, a disease, disorder, or pathological condition is associated with MAPT if the presence, level, activity, and / or form and / or products of MAPT (e.g., transcripts, encoded proteins, etc.) correlate with the incidence and / or susceptibility to that disease, disorder, or pathological condition (e.g., in the entire relevant population). In some embodiments, MAPT-associated pathological conditions, disorders, or diseases may be treated and / or prevented by reducing the expression, level, and / or activity of MAPT transcripts and / or proteins.

[0085] Various MAPT-related conditions, disorders, or diseases have been reported. In some embodiments, the MAPT-related condition, disorder, or disease is Alzheimer's disease (AD). In some embodiments, the MAPT-related condition, disorder, or disease is frontotemporal dementia (FTD).

[0086] In particular, the technologies provided are useful for the treatment or prevention of MAPT-related conditions, disorders, or diseases, such as Alzheimer's disease (AD) and frontotemporal dementia (FTD). In some embodiments, this disclosure relates to the use of MAPT oligonucleotides or compositions thereof in the treatment of MAPT-related disorders, diseases, or conditions, such as Alzheimer's disease (AD) and frontotemporal dementia (FTD).

[0087] The Alzheimer's Association estimates that one in ten people aged 65 and older have Alzheimer's disease (AD), and that nearly six million people in the United States may be affected. According to the report, AD leads to progressive cognitive decline, loss of independence, and eventual death. AD is reported to be characterized by extracellular amyloid plaques and intracellular accumulation of tau aggregates. In addition to AD, tau has also been reported to be involved in the pathophysiology of frontotemporal dementia (FTD). FTD is reported to be a hybrid group of disorders with a prevalence of approximately 20 per 100,000 people. FTD is characterized by degeneration of the frontal, temporal, and other cortical regions, as well as the basal ganglia, thalamus, and other areas, and nearly half of cases are reported to be associated with tau lesions. Subtypes of FTD associated with tau lesions include behavioral FTD (bvFTD), characterized by neuropsychiatric symptoms, and non-fluent primary progressive aphasia (nfvPPA), characterized by speech impairment and word-finding difficulties. Sometimes, motor syndromes, including corticobasal degeneration (CBD) and primary progressive aphasia (PSP), are also reported to be included. Furthermore, in contrast to Alzheimer's disease (AD), which primarily affects the elderly, FTD typically occurs at a younger age, and is therefore more likely to affect people who are still working, and like AD, it can ultimately be fatal. It is being done.

[0088] While we do not wish to be constrained by any theory, it should be noted that several reports have provided genetic and histological evidence (in hereditary and sporadic disorders) regarding the importance of tau in the development of AD and FTD lesions, physical disability, and mortality. In some cases, MAPT has been reported to be genome-wide meaningful for AD itself, but meaningful for shared AD / PD risk (Desikan et al., 2015). Based primarily on family-based genetic studies, MAPT has been clearly demonstrated to be causative in morphologies of FTD with underlying tau lesions (Greaves and Rohrer, 2019). In some cases, tau lesions have been reported to be very well established.

[0089] Tau is reported to be a neural scaffold protein that aggregates within cells during disease, forming neurofibrillary tangles (NFTs), which are key, and reportedly defining, lesions in Alzheimer's disease (AD). Tau lesions can spread from one neuron to the next via prion-like mechanisms, and NFTs are typically found in pyramidal neurons of layers III and V of the hippocampus, entorhinal cortex, and isocortex, while interneurons remain largely unaffected (Braak et al., 2016). Furthermore, tau isolated from AD brains has been reported to be pathogenic when injected into rodent brains (Goedert et al., 2017).

[0090] In some embodiments, the provided techniques (e.g., oligonucleotides, compositions, methods, etc.) reduce the expression, level, function, and / or activity of tau transcripts and the proteins encoded by them. In some embodiments, such reduction addresses intracellular aggregation. In some embodiments, such reduction addresses tau spreading. In some embodiments, such reduction addresses both intracellular aggregation and tau spreading.

[0091] In some embodiments, the Disclosure provides a method for reducing the level, function, and / or activity of tau protein, comprising contacting the protein with an oligonucleotide or composition of the Disclosure. In some embodiments, the Disclosure provides a method for reducing the intracellular aggregation of tau, comprising contacting the protein with an oligonucleotide or composition of the Disclosure. In some embodiments, the Disclosure provides a method for reducing the spreading of tau, comprising contacting the protein with an oligonucleotide or composition of the Disclosure. In some embodiments, the Disclosure provides a method for reducing the intracellular aggregation and spreading of tau, comprising contacting the protein with an oligonucleotide or composition of the Disclosure. In some embodiments, the Disclosure provides a method for reducing the level, function, and / or activity of tau protein in a system, comprising administering an effective amount of an oligonucleotide or composition of the Disclosure to the system. In some embodiments, the Disclosure provides a method for reducing the intracellular aggregation of tau in a system, comprising administering an effective amount of an oligonucleotide or composition of the Disclosure to the system. In some embodiments, the Disclosure provides a method for reducing the spread of tau within a system, comprising administering an effective amount of the oligonucleotide or composition of the Disclosure to the system. In some embodiments, the Disclosure provides a method for reducing intracellular aggregation and spread of tau within a system, comprising administering an effective amount of the oligonucleotide or composition of the Disclosure to the system. In some embodiments, the system is in vitro. In some embodiments, the system is in vivo. In some embodiments, the system is cells or includes cells. In some embodiments, the system is tissue or includes tissue. In some embodiments, the system is an organ or includes organ. In some embodiments, the system is a sample or includes a sample. In some embodiments, the system is a subject or includes a subject. In some embodiments, the system is a mouse or includes a mouse. In some embodiments The system is a non-human primate or includes one. In some embodiments, the system is a human or includes one.

[0092] In some embodiments, treatment or prevention by the provided technology reduces the rate of tau production, thereby reducing, stopping, or reversing the accumulation and further spread of aggregates. In some embodiments, treatment or prevention by the provided technology reduces the rate of clinical decline, or delays or prevents the onset of pathology, impairment, or disease.

[0093] As those skilled in the art will understand, the present disclosure may utilize such pathological conditions, disorders or disease mechanisms, genotypes, symptoms, biomarkers, etc., to characterize / evaluate the technologies provided.

[0094] oligonucleotides In particular, this disclosure provides oligonucleotides of various designs, including various nucleic acid bases and their patterns, sugars and their patterns, internucleotide bonds and their patterns, and / or additional chemical parts and their patterns, as described herein. In some embodiments, the MAPT oligonucleotides provided can lead to a decrease in the expression, level, and / or activity of one or more MAPT genes and / or their products (e.g., transcripts, mRNA, proteins, etc.). In some embodiments, the MAPT oligonucleotides provided can lead to a decrease in the expression, level, and / or activity of one or more MAPT genes and / or their products in cells of a subject or patient. In some embodiments, cells typically express MAPT or produce MAPT proteins. In some embodiments, the provided MAPT oligonucleotides can cause the expression, level, and / or activity of a MAPT target gene or gene product to decrease, and consist of, include, or have a nucleotide sequence comprising, a portion thereof (e.g., a span of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more adjacent nucleotides) of the MAPT oligonucleotides disclosed herein, and the oligonucleotides include at least one modification of a nucleotide, sugar, and / or internucleotide bond that does not exist in nature.

[0095] In some embodiments, MAPT can lead to a decrease in the expression, level, and / or activity of a target gene, such as a MAPT target gene, or its product. In some embodiments, MAPT oligonucleotides can lead to a decrease in the expression, level, and / or activity of a MAPT target gene or its product by RNase H-mediated knockdown. In some embodiments, MAPT oligonucleotides can lead to a decrease in the expression, level, and / or activity of a MAPT target gene or its product by sterically blocking translation after binding to MAPT target gene mRNA, and / or by altering or interfering with mRNA splicing. However, despite this, the disclosure is not limited to any particular mechanism. In some embodiments, the disclosure provides oligonucleotides, compositions, methods, etc., having the ability to function by double-stranded RNA interference, single-stranded RNA interference, RNase H-mediated knockdown, steric hindrance of translation, or a combination of two or more such mechanisms.

[0096] In some embodiments, MAPT oligonucleotides have the ability to mediate a decrease in MAPT expression, level, and / or activity. In some embodiments, MAPT oligonucleotides have the ability to mediate a decrease in tau protein levels. In some embodiments, MAPT oligonucleotides have the ability to mediate a decrease in tau protein levels. In some embodiments, MAPT oligonucleotides have the ability to mediate a decrease in tau aggregation levels, for example, in neurons. In some embodiments, MAPT oligonucleotides have the ability to mediate a decrease in tau spreading levels, for example, from one neuron to another.

[0097] In some embodiments, MAPT oligonucleotides have the ability to mediate a decrease in MAPT expression, level, and / or activity through mechanisms involving mRNA degradation and / or steric hindrance to the translation of MAPT mRNA.

[0098] In some embodiments, the MAPT oligonucleotide has the ability to mediate a decrease in the expression, level, and / or activity of two or more MAPT alleles.

[0099] In some embodiments, the present disclosure relates to a method of treating a MAPT-related disease, disorder, or condition in which MAPT is overexpressed, the method comprising administering a therapeutically effective amount of a MAPT oligonucleotide having the ability to mediate a decrease in the expression, level, and / or activity of MAPT. In some embodiments, there may be multiple forms, e.g., alleles, of MAPT, and the technology provided can reduce the expression, level, and / or activity of two or more or all of such forms and their products.

[0100] In some embodiments, the present disclosure relates to a method of treating a MAPT-related disease, disorder, or condition, the method comprising administering a therapeutic amount of a MAPT oligonucleotide having the ability to mediate a decrease in the expression, level, and / or activity of MAPT.

[0101] In some embodiments, the MAPT oligonucleotide has the ability to mediate a decrease in the expression, level, and / or activity of MAPT through a mechanism involving splicing regulation, such as exon skipping.

[0102] In some embodiments, the MAPT oligonucleotide comprises a structural element or a portion thereof described, for example, in a table herein. In some embodiments, the MAPT oligonucleotide comprises a base sequence (or a portion thereof) described herein (where each T can be independently replaced by U, and vice versa), a chemical modification or chemical modification pattern (or a portion thereof), and / or a format or a portion thereof described herein. In some embodiments, the MAPT oligonucleotide has a base sequence comprising a base sequence of an oligonucleotide disclosed, for example, in a table herein or otherwise disclosed herein (where each T can be independently replaced by U), a chemical modification pattern (or a portion thereof), and / or a format. In some embodiments, such an oligonucleotide, for example, a MAPT oligonucleotide, reduces the expression, level and / or activity of a gene, for example, the MAPT gene, or its gene product.

[0103] In particular, MAPT oligonucleotides can hybridize to their target nucleic acids (e.g., pre-mRNA, mature mRNA, etc.). For example, in some embodiments, MAPT oligonucleotides can hybridize to MAPT nucleic acids derived from a DNA strand (either strand of the MAPT gene). In some embodiments, MAPT oligonucleotides can hybridize to MAPT transcripts. In some embodiments, MAPT oligonucleotides can hybridize to MAPT nucleic acids at any stage of RNA processing, including, but not limited to, pre-mRNA or mature mRNA. In some embodiments, MAPT oligonucleotides can hybridize to any element of MAPT nucleic acids or their complements, including, but not limited to, promoter regions, enhancer regions, transcription termination regions, translation start signals, translation termination codons, coding regions, non-coding regions, exons, introns, intron / exon or exon / intron junctions, 5'UTR, or 3'UTR. In some embodiments, MAPT oligonucleotides can hybridize to their targets as long as there are no more than two mismatches. In some embodiments, MAPT oligonucleotides can hybridize to their target if there is no more than one mismatch. In some embodiments, MAPT oligonucleotides can hybridize to their target if there are no mismatches (for example, when all are CG and / or AT / U base paired).

[0104] In some embodiments, an oligonucleotide can hybridize to two or more variants of the transcript. In some embodiments, a MAPT oligonucleotide can hybridize to two or more or all variants of the MAPT transcript. In some embodiments, a MAPT oligonucleotide can hybridize to two or more or all variants of the MAPT transcript derived from the sense strand.

[0105] In some embodiments, the MAPT target of the MAPT oligonucleotide is a MAPT RNA that is not mRNA.

[0106] In some embodiments, oligonucleotides, e.g., MAPT oligonucleotides, contain increased levels of one or more isotopes. In some embodiments, oligonucleotides, e.g., MAPT oligonucleotides, are labeled with one or more isotopes of one or more elements, e.g., hydrogen, carbon, nitrogen. In some embodiments, oligonucleotides in the provided composition, e.g., MAPT oligonucleotides, e.g., oligonucleotides of multiple compositions, include base modifications, sugar modifications and / or internucleotide bond modifications, where the oligonucleotide contains enhanced levels of deuterium. In some embodiments, oligonucleotides, e.g., MAPT oligonucleotides, contain deuterium at one or more positions (- 1 H 2 Labeled (by replacing H). In some embodiments, one or more of the oligonucleotide chain or any portion conjugated to the oligonucleotide chain (e.g., targeting portion). 1 H 2 It is substituted with H. Such oligonucleotides can be used in any of the compositions and methods described herein.

[0107] In some embodiments, this disclosure 1) Having a common nucleotide sequence complementary to the target sequence in the transcript (e.g., MAPT target sequence); and 2) To provide an oligonucleotide composition comprising multiple oligonucleotides, each containing one or more modified sugar moieties and / or modified internucleotide bonds.

[0108] In some embodiments, MAPT oligonucleotides having a common base sequence may have the same pattern of nucleoside modifications, such as sugar modifications or base modifications. In some embodiments, the nucleoside modification pattern may be represented by a combination of position and modification. In some embodiments, the skeletal bonding pattern includes the position and type of each internucleotide bond (e.g., phosphate, phosphorothioate, substituted phosphorothioate, etc.).

[0109] In some embodiments, for example, in a provided composition, the oligonucleotides in multiple oligonucleotides are of the same oligonucleotide type. In some embodiments, oligonucleotides of a certain oligonucleotide type have a common sugar modification pattern. In some embodiments, oligonucleotides of a certain oligonucleotide type have a common base modification pattern. In some embodiments, oligonucleotides of a certain oligonucleotide type have a common nucleoside modification pattern. In some embodiments, oligonucleotides of a certain oligonucleotide type have the same chemical composition. In some embodiments, oligonucleotides of a certain oligonucleotide type are identical. In some embodiments, oligonucleotides in multiple oligonucleotides are identical. In some embodiments, oligonucleotides in multiple oligonucleotides share the same chemical composition.

[0110] In some embodiments, as illustrated herein, the MAPT oligonucleotide is chiral-controlled by including one or more chiral-controlled internucleotide bonds. In some embodiments, the MAPT oligonucleotide is stereochemically pure. In some embodiments, the MAPT oligonucleotide is substantially separated from other stereoisomers.

[0111] In some embodiments, the MAPT oligonucleotide comprises one or more modified nucleic acid bases, one or more modified sugars, and / or one or more modified internucleotide bonds.

[0112] In some embodiments, the MAPT oligonucleotide comprises one or more modified sugars. In some embodiments, the oligonucleotide of the Disclosure comprises one or more modified nucleic acid bases. Various modifications can be introduced to the sugars and / or nucleic acid bases of the Disclosure. For example, in some embodiments, the modifications are those described in U.S. Patent No. 9006198. In some embodiments, the modifications are U.S. Patent No. 9394333, U.S. Patent No. 9744183, U.S. Patent No. 9605019, U.S. Patent No. 9598458, U.S. Patent No. 9982257, U.S. Patent Application Publication No. 10160969, U.S. Patent Application Publication No. 10479995, U.S. Patent Application Publication No. 2020 / 0056173, U.S. Patent Application Publication No. 2018 / 0216107, U.S. Patent Application Publication No. 2019 / 0127733, U.S. Patent Application Publication No. 10450568, U.S. Patent Application Publication No. 2019 / 0077817, U.S. Patent Application Publication No. 2019 / 0249173, U.S. Patent Application Publication No. 2019 / The modifications are those described in International Publication No. 0375774, International Publication No. 2018 / 223056, International Publication No. 2018 / 223073, International Publication No. 2018 / 223081, International Publication No. 2018 / 237194, International Publication No. 2019 / 032607, International Publication No. 2019 / 055951, International Publication No. 2019 / 075357, International Publication No. 2019 / 200185, International Publication No. 2019 / 217784, International Publication No. 2019 / 032612 and / or International Publication No. 2020 / 191252 (each of these sugar, base and internucleotide linkage modifications is independently incorporated herein by reference).

[0113] When used in this disclosure, in some embodiments, “one or more” means 1 to 200, 1 to 150, 1 to 100, 1 to 90, 1 to 80, 1 to 70, 1 to 60, 1 to 50, 1 to 40, 1 to 30 or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25. In some embodiments, “one or more” is 1. In some embodiments, “one or more” is 2. In some embodiments, “one or more” is 3. In some embodiments, “one or more” is 4. In some embodiments, “one or more” is 5. In some embodiments, “one or more” is 6. In some embodiments, “one or more” is 7. In some embodiments, “one or more” is 8. In some embodiments, “one or more” is 9. In some embodiments, “one or more” is 10. In some embodiments, “one or more” is at least 1. In some embodiments, "1 or more" means at least 2. In some embodiments, "1 or more" means at least 3. In some embodiments, "1 or more" means at least 4. In some embodiments, "1 or more" means at least 5. In some embodiments, "1 or more" means at least 6. In some embodiments, "1 or more" means at least 7. In some embodiments, "1 or more" means at least 8. In some embodiments, "1 or more" means at least 9. In some embodiments, "1 or more" means at least 10.

[0114] When used in this disclosure, in some embodiments, “at least 1” means 1-200, 1-150, 1-100, 1-90, 1-80, 1-70, 1-60, 1-50, 1-40, 1-30 or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 1 3, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, "at least 1" is 1. In some embodiments, "at least 1" is 2. In some embodiments, "at least 1" is 3. In some embodiments, "at least 1" is 4. In some embodiments, "at least 1" is 5. In some embodiments, "at least 1" is 6. In some embodiments, "at least 1" is 7. In some embodiments, "at least 1" is 8. In some embodiments, "at least 1" is 9. In some embodiments, "at least 1" is 10.

[0115] In some embodiments, the MAPT oligonucleotide is or includes one of the MAPT oligonucleotides listed in the table.

[0116] As demonstrated in this disclosure, in some embodiments, the provided oligonucleotide (e.g., MAPT oligonucleotide) is characterized by knocking down its target (e.g., a MAPT transcript relative to a MAPT oligonucleotide) when it comes into contact with the transcript in a knockdown system.

[0117] In some embodiments, the oligonucleotide is provided in salt form. In some embodiments, the oligonucleotide is provided as a salt containing negatively charged internucleotide bonds (e.g., phosphorothioate internucleotide bonds, native phosphate bonds, etc.) present in its salt form. In some embodiments, the oligonucleotide is provided as a pharmaceutically acceptable salt. In some embodiments, the oligonucleotide is provided as a metal salt. In some embodiments, the oligonucleotide is provided as a sodium salt. In some embodiments, the oligonucleotide is provided as a metal salt, for example, a sodium salt, where each negatively charged internucleotide bond is independently in salt form (e.g., for the sodium salt, -OP(O)(SNa)-O- for the phosphorothioate internucleotide bond, -OP(O)(ONa)-O- for the native phosphate bond, etc.).

[0118] Base sequence In some embodiments, the MAPT oligonucleotide comprises the nucleotide sequence described herein or a portion thereof having 0 to 5 (e.g., 0, 1, 2, 3, 4, or 5) mismatches (e.g., 5 to 50, 5 to 40, 5 to 30, 5 to 20, or 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or at least 10 or at least 15 adjacent nucleic acid base spans) (wherein each T may be independently substituted with U, and vice versa). In some embodiments, the MAPT oligonucleotide comprises the nucleotide sequence described herein or a portion thereof, wherein the portion is at least 10 adjacent nucleic acid base spans, or at least 15 adjacent nucleic acid base spans having 1 to 5 mismatches. In some embodiments, the MAPT oligonucleotide comprises a nucleotide sequence or a portion thereof as described herein, wherein the portion is a span of at least 10 adjacent nucleic acid bases, or a span of at least 10 adjacent nucleic acid bases having 1 to 5 mismatches (wherein each T may be independently substituted with U, and vice versa). In some embodiments, the oligonucleotide sequence is 10 to 50 (e.g., about or at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45; in some embodiments, at least 15; in some embodiments, at least 16; in some embodiments, at least 17; in some embodiments, at least 18; in some embodiments, at least 19; in some embodiments, at least 20; in some embodiments In its morphology, it contains or comprises at least 21 adjacent bases; in some embodiments, at least 22; in some embodiments, at least 23; in some embodiments, at least 24; and in some embodiments, at least 25.

[0119] As those skilled in the art will understand, the nucleotide sequences of MAPT oligonucleotides are typically of sufficient length to mediate target-specific knockdown and complementary to their targets, such as RNA transcripts (e.g., pre-mRNA, mature mRNA, etc.). In some embodiments, the nucleotide sequences of MAPT oligonucleotides are of sufficient length to mediate target-specific knockdown and identical to MAPT transcript targets. In some embodiments, MAPT oligonucleotides are complementary to a portion of a MAPT transcript (MAPT transcript target sequence). In some embodiments, the nucleotide sequences of MAPT oligonucleotides are 90% or more identical to the nucleotide sequences of oligonucleotides disclosed in the table (wherein each T can be independently substituted with U, and vice versa). In some embodiments, the nucleotide sequences of MAPT oligonucleotides are 95% or more identical to the nucleotide sequences of oligonucleotides disclosed in the table (wherein each T can be independently substituted with U, and vice versa). In some embodiments, the base sequence of the MAPT oligonucleotide includes 15 or more adjacent spans of the oligonucleotides disclosed in the table (wherein each T can be independently substituted with U, and vice versa), except in cases where one or more bases within a span are absent (e.g., a nucleic acid base is not present in the nucleotide). In some embodiments, the base sequence of the MAPT oligonucleotide includes 19 or more adjacent spans of the MAPT oligonucleotides disclosed herein, except in cases where one or more bases within a span are absent (e.g., a nucleic acid base is not present in the nucleotide). In some embodiments, the base sequence of the MAPT oligonucleotide includes 19 or more adjacent spans of the oligonucleotides disclosed herein (wherein each T can be independently substituted with U, and vice versa), except for a difference of one or two bases at the 5' and / or 3' ends of the base sequence.

[0120] In some embodiments, the nucleotide sequence of the oligonucleotide is ACGTTGCAGTGTTCCACUAU, ACTATCCTCCTTCAGCUCCU, AGTGTTCCACTATCCUCCUU, ATCCTCCTTCAGCTCCUGCA, CACGTTGCAGTGTTCCACUA, CACTATCCTCCTTCAGCUCC, CAGTGTTCCACTATCCUCCU, CCACGTTGCAGTGTTCCACU, CCACTATCCTCCTTCAGCUC, CGTTGCAGTGTTCCACUAUC, CTATCCTCCTTCAGCUCCUG, GCAGTGTTCCACTATCCUCC, GTGTCC ACTATCCTCCUUC, GTTCCACTATCCTCCUUCAG, GUGUUCCACTATCCTCCTTC, TATCCTCCTTCAGCTCCUGC, TCCACTATCCTCCTTCAGCU, TGCAGTGTTCCACTAUCCUC, TGTTCCACTATCCTCCUUCA, TTCCACTATCCTCCTUCAGC, or TTGCAGTGTTCCACTAUCCU (wherein each T can be independently replaced by U, and vice versa), or containing the same, or containing 10 to 20 thereof, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 adjacent bases. In some embodiments, the nucleotide sequence of the oligonucleotide is such a sequence (wherein each T can be independently replaced by U, and vice versa) or contains the same. In some embodiments, the nucleotide sequence of the oligonucleotide is such a sequence (wherein each T can be independently replaced by U, and vice versa). In some embodiments, the base sequence of the polypeptide is ACGTTGCAGTGTTCCACUAU, ACTATCCTCCTTCAGCUCCU, AGTGTTCCACTATCCUCCUU, ATCCTCCTTCAGCTCCUGCA, CACGTTGCAGTGTTCCACUA, CACTATCCTCCTTCAGCUCC, CAGTGTTCCACTATCCUCCUCU, CCACGTTGCAGTGTTCCA Is CU, CCACTATCCTCCTTCAGCUC, CGTTGCAGTGTTCCACUAUC, CTATCCTCCTTCAGCUCCUG, GCAGTGTTCCACTATCCUCC, GTGTTCCACTATCCTCCUUC, GTTCCACTATCCTCCUUCAG, GUGUUCCACTATCCTCCTTC, TATCCTCCTTCAGCTCCUGC, TCCACTATCCTCCTTCAGCU, TGCAGTGTTCCACTAUCCUC, TGTTCCACTATCCTCCUUCA, TTCCACTATCCTCCTUCAGC or TTGCAGTGTTCCACTAUCCU, or contains it, or contains 10 to 20, for example 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 adjacent bases thereof. In some embodiments, the nucleotide sequence of the oligonucleotide is such a sequence or contains it. In some embodiments, the nucleotide sequence of the oligonucleotide is such a sequence.

[0121] In some embodiments, the nucleotide sequence of the oligonucleotide is GTGTTCCACTATCCTCCUUC (wherein each T can be independently replaced by U, and vice versa), contains the same, or contains 10 to 20 of the same, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 adjacent nucleotides. In some embodiments, the nucleotide sequence of the oligonucleotide is GTGTTCCACTATCCTCCUUC (wherein each T can be independently replaced by U, and vice versa), or contains the same. In some embodiments, the nucleotide sequence of the oligonucleotide is GTGTTCCACTATCCTCCUUC (wherein each T can be independently replaced by U, and vice versa). In some embodiments, the nucleotide sequence of the oligonucleotide is GTGTTCCACTATCCTCCUUC, or contains thereto, or contains 10 to 20 adjacent nucleotides, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In some embodiments, the nucleotide sequence of the oligonucleotide is GTGTTCCACTATCCTCCUUC, or contains thereto. In some embodiments, the nucleotide sequence of the oligonucleotide is GTGTTCCACTATCCTCCUUC.

[0122] In some embodiments, the present disclosure relates to oligonucleotides having a nucleotide sequence comprising any oligonucleotide disclosed herein (wherein each T can be independently replaced by U, and vice versa).

[0123] In some embodiments, the present disclosure relates to oligonucleotides having a nucleotide sequence comprising at least 15 adjacent nucleotides of any oligonucleotide sequence disclosed herein (wherein each T may be independently replaced by U, and vice versa).

[0124] In some embodiments, the present disclosure relates to oligonucleotides having a nucleotide sequence that is at least 90% identical to the nucleotide sequence of any oligonucleotide disclosed herein (wherein each T can be independently replaced by U, and vice versa).

[0125] In some embodiments, the present disclosure relates to an oligonucleotide having a nucleotide sequence that is at least 95% identical to the nucleotide sequence of any oligonucleotide disclosed herein (wherein each T can be independently replaced by U, and vice versa).

[0126] In some embodiments, the nucleotide sequence of the oligonucleotide is the nucleotide sequence of any oligonucleotide described herein (wherein each T can be independently replaced by U, and vice versa), contains the same, or contains 10 to 20 of the same, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 adjacent nucleotides.

[0127] In some embodiments, the MAPT oligonucleotide is selected from Table 1.

[0128] In some embodiments, the nucleotide sequence of the MAPT oligonucleotide is complementary to the MAPT transcript or a portion thereof.

[0129] In some embodiments, the base sequence of a MAPT oligonucleotide is complementary to a portion of a MAPT nucleic acid sequence, such as a MAPT gene sequence, MAPT transcript, or MAPT mRNA sequence. In some embodiments, the MAPT oligonucleotide is identical to a portion of a MAPT nucleic acid sequence, such as a MAPT gene sequence, MAPT transcript, or MAPT mRNA sequence. In some embodiments, the portion consists of or contains 10 or more adjacent nucleic acid bases, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more. In some embodiments, it is 15 or more. In some embodiments, it is 16 or more. In some embodiments, it is 17 or more. In some embodiments, it is 18 or more. In some embodiments, it is 19 or more. In some embodiments, it is 20 or more. In some embodiments, the base sequence of such a portion is characteristic of MAPT in that no other genome sequence or transcript sequence containing the same sequence as that portion exists in the system. In some embodiments, there are no other genome sequences or transcript sequences in the system that contain a portion differing from only one or fewer nucleic acid bases. In some embodiments, there are no other genome sequences or transcript sequences in the system that contain a portion differing from only two or fewer nucleic acid bases. In some embodiments, the portion of a gene complementary to an oligonucleotide is referred to as the target sequence of the oligonucleotide. In some embodiments, the system is or includes a cell, sample, tissue, organ, or species. For example, for an oligonucleotide targeting human MAPT, the relevant species in many embodiments is human. In some embodiments, when, for example, interspecies activity and / or properties are characterized and / or evaluated, the system may be or include multiple species. In some embodiments, such portion is in an exon. In some embodiments, such portion is in an intron. In some embodiments, such portion is in intron 11. In some embodiments, such portion spans both an intron and an exon. In some embodiments, such portion spans two exons.In some embodiments, such portion is located in the 5'-UTR region. In some embodiments, such portion is located in the 3'-UTR region.

[0130] In some embodiments, MAPT oligonucleotides target two or more or all alleles of MAPT (when multiple alleles are present in the relevant system). In some embodiments, the oligonucleotides reduce the expression, level, and / or activity of both wild-type MAPT and mutant MAPT, and / or their transcripts and / or products.

[0131] In some embodiments, the nucleotide sequences of the provided oligonucleotides are fully complementary to MAPT target sequences in both human and non-human primates (NHPs). In some embodiments, such sequences may be particularly useful because they can be readily evaluated in both human and non-human primates.

[0132] In some embodiments, the MAPT oligonucleotide comprises a nucleotide sequence or portion thereof as listed in the table (wherein each T can be independently replaced by U, and vice versa), and / or a sugar, nucleic acid base and / or internucleotide bond modification and / or pattern thereof as listed in the table, and / or additional chemical portions as listed in the table (added to the oligonucleotide chain, e.g., a target portion, a lipid portion, a carbohydrate portion, etc.).

[0133] In some embodiments, the terms “complementary,” “fully complementary,” and “substantially complementary” mean oligonucleotides (for example) as those skilled in the art would understand from the context of their use. For example, MAPT oligonucleotides can be used in relation to the degree of base match between a MAPT oligonucleotide sequence and a target sequence (e.g., a MAPT target sequence). As a non-limiting example, if the target sequence has the base sequence 5'-GCAUAGCGAGCGAGGGAAAAC-3', then an oligonucleotide with the base sequence 5'GUUUUCCCUCGCUCGCUAUGC-3' is complementary (perfectly complementary) to such a target sequence. It should be noted that substitution of U with T, or vice versa, generally does not change the magnitude of complementarity. When used herein, an oligonucleotide that is "substantially complementary" to a target sequence is mostly or almost complementary, but not 100% complementary. In some embodiments, a substantially complementary sequence (e.g., a MAPT oligonucleotide) has 1, 2, 3, 4, or 5 mismatches when aligned with its target sequence. In some embodiments, a MAPT oligonucleotide has a base sequence that is substantially complementary to a MAPT target sequence. In some embodiments, the MAPT oligonucleotide has a nucleotide sequence substantially complementary to the complementary sequence of the MAPT oligonucleotide disclosed herein. As those skilled in the art will understand, in some embodiments, the oligonucleotide sequence does not need to be 100% complementary to its target for the oligonucleotide to perform its function (e.g., knockdown of the target nucleic acid). Typically, when determining complementarity, A and T (or U) are complementary nucleic acid bases, and C and G are complementary nucleic acid bases.

[0134] In some embodiments, the disclosure provides MAPT oligonucleotides comprising sequences found in the oligonucleotides listed in the table. In some embodiments, the disclosure provides MAPT oligonucleotides comprising sequences found in the oligonucleotides listed in the table (wherein one or more Us are independently and optionally substituted with T, and vice versa). In some embodiments, the MAPT oligonucleotide may comprise at least one T and / or at least one U. In some embodiments, the disclosure provides MAPT oligonucleotides comprising sequences found in the oligonucleotides listed in the table, wherein the sequences have more than 50% identity with the sequences of the oligonucleotides listed in the table. In some embodiments, the disclosure provides MAPT oligonucleotides comprising sequences of the oligonucleotides disclosed in the table. In some embodiments, the disclosure provides MAPT oligonucleotides of a nucleotide sequence which is the sequence of the oligonucleotides disclosed in the table (wherein each T may be independently substituted with U, and vice versa). In some embodiments, the Disclosure provides MAPT oligonucleotides comprising sequences found in the oligonucleotides in the Table, wherein the oligonucleotide has a skeletal bonding pattern, a skeletal chiral center pattern, and / or skeletal phosphorus modification pattern of the same or a different oligonucleotide in the Table herein.

[0135] In particular, this disclosure presents various oligonucleotides having the nucleotide sequences defined herein in Table 1A and elsewhere. In some embodiments, this disclosure provides oligonucleotides having the nucleotide sequences of, or including, the oligonucleotides disclosed herein, e.g., in Table 1, e.g. (wherein each T can be independently replaced by U, and vice versa), or a nucleotide sequence containing a portion thereof. In some embodiments, this disclosure provides oligonucleotides having the nucleotide sequences of, or including, the oligonucleotides disclosed herein, e.g., in Table 1, e.g. (wherein each T can be independently replaced by U, and vice versa), where the oligonucleotide further includes chemical modifications, stereochemistry, format, additional chemical parts described herein (e.g., targeting parts, lipid parts, carbohydrate parts, etc.), and / or other structural features.

[0136] In some embodiments, “a portion” (for example, a portion of a nucleotide sequence or modification pattern) is at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 monomer unit lengths (for example, for a nucleotide sequence, at least 5, 6, (7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 bases long). In some embodiments, a "part" of the base sequence is at least 5 bases long. In some embodiments, a "part" of the base sequence is at least 10 bases long. In some embodiments, a "part" of the base sequence is at least 15 bases long. In some embodiments, a "part" of the base sequence is at least 16, 17, 18, 19, or 20 bases long. In some embodiments, a "part" of the base sequence is at least 20 bases long. In some embodiments, a part of the base sequence consists of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or more adjacent (consecutive) bases. In some embodiments, a part of the base sequence consists of 15 or more adjacent (consecutive) bases. In some embodiments, a portion of the nucleotide sequence consists of 16, 17, 18, 19, or 20 or more adjacent (consecutive) nucleotides.

[0137] In some embodiments, the Disclosure provides oligonucleotides (e.g., MAPT oligonucleotides) whose nucleotide sequence is the nucleotide sequence or a portion thereof of the oligonucleotides in the Table, where each T may be independently replaced by U, and vice versa. In some embodiments, the Disclosure provides MAPT oligonucleotides of the sequences of the oligonucleotides in the Table, where the oligonucleotide has the ability to lead to a decrease in the expression, level, and / or activity of the MAPT gene or its gene product. As those skilled in the art will understand, in the provided sequences, each U may be optionally and independently replaced by T, and vice versa, and sequences may include mixtures of U and T. In some embodiments, C may be optionally and independently replaced by 5mC.

[0138] In some embodiments, a portion is a span of at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 total nucleotides containing 0 to 3 mismatches. In some embodiments, a portion is a span of at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 total nucleotides containing 0 to 3 mismatches, where a span with 0 mismatches is complementary, and a span with 1 or more mismatches is an unrestricted example of substantial complementarity. In some embodiments, the bases include a portion that is characteristic of nucleic acids (e.g., genes) in that a portion of it is identical or complementary to a portion of the nucleic acid or its transcript, but is not identical or complementary to any other nucleic acid (e.g., genes) or its transcript in the same genome. In some embodiments, a portion of it is characteristic of human MAPT.

[0139] In some embodiments, the oligonucleotides provided, e.g., MAPT oligonucleotides, have a total nucleotide length of about 49, 45, 40, 30, 35, 25, or 23 or fewer, as described herein. In some embodiments, if the 5' end of the sequence described herein begins with U or T, U may be deleted and / or replaced with another base. In some embodiments, the oligonucleotide has a nucleotide sequence that is or contains, or contains, a nucleotide sequence of the oligonucleotide in the table, in the format or a portion of the format disclosed herein, where each T may be independently replaced with U, and vice versa.

[0140] In some embodiments, the oligonucleotide, for example, MAPT oligonucleotide, is sterically random. In some embodiments, the MAPT oligonucleotide is chiral controlled. In some embodiments, the MAPT oligonucleotide is Chiral-pure (or "stereo-pure," "stereochemically pure"), where the oligonucleotide exists as a single stereoisomer (often as a single diastereoisomer, since multiple chiral centers can exist in the oligonucleotide, e.g., at the bound phosphorus, sugar carbon). As those skilled in the art will understand, chiral-pure oligonucleotides are separated from other stereoisomers (some impurities may exist, as chemical and biological processes, selectivity, and / or purification rarely, if not never, reach absolute perfection). In chiral-pure oligonucleotides, each chiral center is independently defined with respect to its configuration (for chiral-pure oligonucleotides, each internucleotide bond is independently sterically defined or chiral-controlled). In contrast to chiral-controlled and chiral-pure oligonucleotides containing sterically defined bound phosphorus, a racemic (or "sterically random," "uncontrolled") oligonucleotide containing chiral bound phosphorus from conventional phosphoramidite oligonucleotide synthesis without stereochemical control during the coupling step, for example, combined with conventional sulfurization (which creates sterically random phosphorothioate internucleotide bonds), typically refers to a random mixture of diastereoisomers (or "diastereomers") because there are multiple chiral centers in the oligonucleotide (for example, from conventional oligonucleotide preparations using reagents that do not contain chiral elements other than those in the nucleoside and bound phosphorus). For example, A * A * A [in the formula, * Regarding the phosphorothioate internucleotide bond (which contains chiral phosphorus), the racemic oligonucleotide formulation consists of four diastereomers [2 2 =4, Consider that there are two chiral phosphorus molecules, each of which can exist in one of two configurations (Sp or Rp):A * SA * SA, A * SA * RA, A* RA * SA and A * R A * RA [in the formula, * S represents an Sp phosphorothioate internucleotide bond. * [R represents the Rp phosphorothioate internucleotide bond]. Chiralally pure oligonucleotides, e.g., A * SA * Regarding SA, it exists in a single stereoisomer form, and other stereoisomers (e.g., diastereomer A) * SA * R A keyA * RA * SA and A * RA * It is separated from RA.

[0141] In some embodiments, the MAPT oligonucleotide contains one, two, three, four, five, six, seven, eight, nine, ten or more sterically random internucleotide links (e.g., a mixture of Rp and Sp-linked phosphorus in the internucleotide links from conventional non-chiralized oligonucleotide synthesis). In some embodiments, the MAPT oligonucleotide contains one or more chiralized internucleotide links (e.g., 1-50, 1-40, 1-30, 1-25, 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more) chiralized internucleotide links (e.g., Rp or Sp-linked phosphorus in the internucleotide links from chiralized oligonucleotide synthesis). In some embodiments, the internucleotide links are phosphorothioate internucleotide links. In some embodiments, the internucleotide bond is a sterically random phosphorothioate internucleotide bond. In some embodiments, the internucleotide bond is a chiralally controlled phosphorothioate internucleotide bond.

[0142] In particular, this disclosure provides techniques for preparing chiral-controlled (and in some embodiments, stereochemically pure) oligonucleotides. In some embodiments, the oligonucleotides are stereochemically pure. In some embodiments, the oligonucleotides of this disclosure are available in concentrations of about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 15%, 20%. The purity is %, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%. In some embodiments, the internucleotide bonds of oligonucleotides are one or more (e.g., 1-50, 1-40, 1-30, 1-25, 1-20, 5-50, 5-40, 5-30, 5-25, 5-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more). ) comprises or consists of chiral internucleotide bonds, each independently having a diastereoplecy of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%, typically at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%. In some embodiments, the oligonucleotides of the present disclosure, e.g., MAPT oligonucleotides, are (DS) CILThe diastereopurity is as described herein (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% or more), and CIL is the number of chiral-controlled internucleotide bonds (e.g., 1-50, 1-40, 1-30, 1-25, 1-20, 5-50, 5-40, 5-30, 5-25, 5-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more). In some embodiments, DS is 95%-100%. In some embodiments, each internucleotide bond is independently chiralized, and CIL is the number of chiralized internucleotide bonds.

[0143] As an example, specific MAPT oligonucleotides containing particular exemplary base sequences, nucleic acid base modifications and their patterns, sugar modifications and their patterns, internucleotide bonds and their patterns, linked phosphorus stereochemistry and its patterns, linkers, and / or additional chemical moieties are presented in Tables 1A and 1B below. In particular, by targeting MAPT transcripts using oligonucleotides, for example, those listed in Table 1A, the levels of MAPT transcripts and / or their products may be reduced, for example.

[0144] [Table 1]

[0145] [Table 2]

[0146] [Table 3]

[0147] [Table 4]

[0148] Table 5

[0149] Table 6

[0150] Table 7

[0151] Table 8

[0152] Table 9

[0153] Table 10

[0154] Table 11

[0155] Table 12

[0156] Table 13

[0157] Table 14

[0158] [Table 15]

[0159] [Table 16]

[0160] Note: In Tables 1A and 1B, the descriptions, base sequences, and stereochemistry / bonding are determined by their length. It may be divided into multiple rows. Unless otherwise specified, all oligonucleotides in Tables 1A and 1B are single-stranded. As those skilled in the art will understand, the nucleoside units are unmodified and, unless otherwise indicated (e.g., by r, m, m5, eo, etc.), contain unmodified nucleic acid bases and 2'-deoxy sugars; the bonds are natural phosphate bonds unless otherwise indicated; and acidic / basic groups may exist independently in their salt forms. Parts and modifications in oligonucleotides (or other compounds, e.g., those useful for preparing the oligonucleotides provided, including those parts or modifications): m:2'-OMe; m5: Methyl methyl group at position 5 of C (the nucleic acid base is 5-methylcytosine); m5Ceo:5-methyl 2'-O-methoxyethyl C; eo:2'-MOE(2'-O-methoxyethyl,2'-OCH2CH2OCH3); O,PO: Phosphate diester (phosphate). This can be a terminal group (or a component thereof), or a bond, such as a bond between a linker and an oligonucleotide chain, or an internucleotide bond (natural phosphate bond). Phosphate diesters are typically indicated by "O" in the stereochemistry / bonding column and typically not marked in the description column (if it is a terminal group, e.g., a 5'-terminal group, it is indicated in the description column and typically not indicated in the stereochemistry / bonding column); if a bond is not indicated in the description column, it is typically a phosphate diester unless otherwise specified. Note that phosphate bonds between linkers (e.g., L001) and oligonucleotide chains may not be marked in the description column and may not be indicated by "O" in the stereochemistry / bonding column; *,PS: phosphorothioate. This can be a terminal group (if it is a terminal group, e.g., a 5'-terminal group, it will be indicated in the description and not typically indicated in stereochemistry / bonding), or a bond, e.g., a bond between a linker (e.g., L001) and an oligonucleotide chain, an internucleotide bond (phosphorothioate internucleotide bond), etc. R,Rp:Rp configuration phosphorothioate. Note that *R (or *R) indicates a single phosphorothioate bond in the Rp configuration; S,Sp:Sp configuration phosphorothioate. Note that *S (or *S) in the description indicates a single phosphorothioate bond in the Sp conformation; X: Stereomorphically random phosphorothioates; n001: [ka] ; nX is a three-dimensionally random n001; n001R, nR: n001 in Rp configuration; n001S,nS:n001 in Sp configuration.

[0161] length As those skilled in the art will understand, oligonucleotides can be of various lengths to provide desirable properties and / or activities for various uses. Many techniques for evaluating, selecting and / or optimizing oligonucleotide lengths are available in the art and can be utilized in this disclosure. As demonstrated herein, in many embodiments, MAPT oligonucleotides are of a length suitable for hybridizing with their target to reduce the level of their target and / or encoded product. In some embodiments, oligonucleotides The nucleotide is long enough to recognize the target nucleic acid (e.g., MAPT mRNA). In some embodiments, the oligonucleotide is long enough to distinguish the target nucleic acid from other nucleic acids (e.g., nucleic acids with non-MAPT base sequences) to reduce off-target effects. In some embodiments, the MAPT oligonucleotide is short enough to reduce manufacturing or production complexity and lower product costs.

[0162] In some embodiments, the base sequence of the oligonucleotide is approximately 10 to 500 nucleic acid bases long. In some embodiments, the base sequence is approximately 10 to 500 nucleic acid bases long. In some embodiments, the base sequence is approximately 10 to 50 nucleic acid bases long. In some embodiments, the base sequence is approximately 15 to 50 nucleic acid bases long. In some embodiments, the base sequence is approximately 15 to 30 nucleic acid bases long. In some embodiments, the base sequence is approximately 10 to 25 nucleic acid bases long. In some embodiments, the base sequence is approximately 15 to 22 nucleic acid bases long. In some embodiments, the base sequence is approximately 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleic acid bases long. In some embodiments, the base sequence is approximately 18 nucleic acid bases long. In some embodiments, the base sequence is approximately 19 nucleic acid bases long. In some embodiments, the base sequence is approximately 20 nucleic acid bases long. In some embodiments, the base sequence is approximately 21 nucleic acid bases long. In some embodiments, the nucleotide sequence is approximately 22 nucleic acid bases long. In some embodiments, the nucleotide sequence is approximately 23 nucleic acid bases long. In some embodiments, the nucleotide sequence is approximately 24 nucleic acid bases long. In some embodiments, the nucleotide sequence is approximately 25 nucleic acid bases long. In some embodiments, each nucleic acid base is optionally substituted with A, T, C, G, U, or an optionally substituted tautomer of A, T, C, G, or U.

[0163] Area, wing, and core In some embodiments, oligonucleotides, such as MAPT oligonucleotides, comprise several regions, each independently containing one or more sequential nucleosides and optionally one or more internucleotide bonds. In some embodiments, a region differs from one or more adjacent regions in that it possesses one or more structural features that are different from the corresponding structural features of one or more adjacent regions. Exemplary structural features include nucleic acid base modifications and their patterns, sugar modifications and their patterns, internucleotide bonds and their patterns (which may be internucleotide bond types (e.g., phosphate, phosphorothioate, phosphorothioate triester, neutral internucleotide bond, etc.) and their patterns), binding phosphate modifications (skeletal phosphate modifications) and their patterns, skeletal chiral center (binding phosphate) stereochemistry and its patterns [e.g., combinations of Rp and / or Sp of chiral-controlled internucleotide bonds (in order from 5' to 3'), which, if present, optionally accompany unchiral-controlled internucleotide bonds and / or native phosphate bonds (e.g., OSOOO RSSRS SSSRS SOOOS)]. In some embodiments, a region contains chemical modifications (e.g., sugar modifications, base modifications, internucleotide bonds, or stereochemistry of internucleotide bonds) that are not present in one or more adjacent regions. In some embodiments, a region lacks chemical modifications that are present in one or more adjacent regions.

[0164] In some embodiments, the oligonucleotide comprises or consists of two or more regions. In some embodiments, the oligonucleotide comprises or consists of three or more regions. In some embodiments, the oligonucleotide comprises or consists of two adjacent regions, where one region is referred to as the wing region and the other as the core region. The structure of such an oligonucleotide comprises or consists of a wing-core or core-wing structure. In some embodiments, the oligonucleotide comprises or consists of three adjacent regions, where one region contains two adjacent regions laterally located. In some embodiments, the central region is the core region, and each of the laterally located regions is This is called a wing region (5'-wing if it is bound to the 5' end of the core, or 3'-wing if it is bound to the 3' end of the core). The structure of such oligonucleotides includes or consists of a wing-core-wing structure.

[0165] In some embodiments, the first region (e.g., wing) differs from the second region (e.g., core) in that it contains one or more sugar modifications or patterns thereof that are not present in the second region. In some embodiments, the first (e.g., wing) region contains sugar modifications or patterns thereof that are not present in the second (e.g., core) region. In some embodiments, the sugar modification is a 2'-modification. In some embodiments, the 2'-modification is a 2'-OR, where R is optionally substituted with C. 1~6 It is aliphatic. In some embodiments, the 2'-modification is 2'-OR, where R is optionally substituted with C. 1~6It is alkyl. In some embodiments, the 2'-modification is 2'-MOE. In some embodiments, the 2'-modification is 2'-OMe. In some embodiments, the modified sugar is a bicyclic sugar, e.g., an LNA sugar. In some embodiments, each sugar in a region is independently modified. In some embodiments, each sugar in a region (e.g., a wing) independently includes modifications that may be the same as or different from each other. In some embodiments, each sugar in a region (e.g., a wing) includes the same modification, e.g., a 2'-modification as described herein. In some embodiments, the sugar in a region (e.g., a core) is unmodified. In some embodiments, each sugar in a region (e.g., a core) is an unmodified DNA sugar (having two -H at the 2' position). In some embodiments, the oligonucleotide structure provided includes or comprises a wing-core, core-wing, or wing-core-wing structure, where each wing independently includes one or more sugar modifications, and each sugar in the core is a native DNA sugar (having two -H at the 2' position).

[0166] In addition to or instead of the above, a first region (e.g., a wing) may contain one or more internucleotide bonds or patterns thereof that differ from another region (e.g., a core or another wing). In some embodiments, a region (e.g., a wing) contains two or more consecutive native phosphate bonds. In some embodiments, a region (e.g., a core) does not contain consecutive native phosphate bonds. In some embodiments, the oligonucleotide structure provided comprises or consists of a wing-core, core-wing, or wing-core-wing structure, where at least one wing independently contains two or more consecutive native phosphate bonds, and the core does not contain consecutive native phosphate bonds. In some embodiments, in a wing-core-wing structure, each wing independently contains two or more consecutive internucleotide bonds. Unless otherwise noted, for the purposes of stereochemistry of the wing-core-wing structure, the internucleotide bonds linking the core to the wings are included in the core (see, e.g., above).

[0167] In some embodiments, the region is a 5'-wing, a 3'-wing, or a core. In some embodiments, the 5'-wing is at the 5' end of the oligonucleotide, the 3'-wing is at the 3' end, and the core is between the 5'-wing and the 3'-wing, and the oligonucleotide comprises or comprises a wing-core-wing structure or format. In some embodiments, the core comprises a span of adjacent native DNA sugars (2'-deoxyribose). In some embodiments, the core comprises at least 5 spans of adjacent native DNA sugars (2'-deoxyribose). In some embodiments, the core comprises at least 10 spans of adjacent native DNA sugars (2'-deoxyribose). In some embodiments, the core is referred to as a gap. In some embodiments, an oligonucleotide comprising or comprising a wing-core-wing structure is described as a gapmer. In some embodiments, the structure of the provided oligonucleotide comprises or comprises a wing-core structure. In some embodiments, the structure of the provided oligonucleotide comprises a core-wing structure In some embodiments, the oligonucleotide structure comprises or comprises an oligonucleotide chain comprising or comprising a wing-core-wing, wing-core, or wing-core structure, where the oligonucleotide chain is optionally conjugated to an additional chemical portion via a linker as described herein. In some embodiments, the disclosure provides oligonucleotides that target MAPT and have a structure comprising one or two wings and a core, and oligonucleotides comprising or comprising a wing-core-wing, core-wing, or wing-core structure.

[0168] Ribonuclease H (RNase H, e.g., RNase H1, RNase H2, etc.) is reported to recognize structures containing RNA-DNA hybrids (e.g., heteroduplexes) and cleave the RNA. In some embodiments, oligonucleotides containing spans of adjacent native DNA sugars (e.g., 2'-deoxyribose in the core region) have the ability to anneal to RNA such as mRNA to form a heteroduplex; and this heteroduplex structure has the ability to be recognized by RNase H, and this RNA is cleaved by RNase H. In some embodiments, the core of the oligonucleotide provided contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more adjacent native DNA sugars, and the core has the ability to specifically anneal to a target transcript [e.g., MAPT transcripts (e.g., pre-mRNA, mature mRNA, etc.)]; and the formed structure has the ability to be recognized by RNase H, and this transcript is cleaved by RNase H. In some embodiments, the core of the oligonucleotide provided contains 5 or more adjacent DNA sugars.

[0169] The regions, e.g., wings, cores, etc., can be of various suitable lengths. In some embodiments, the regions (e.g., wings, cores, etc.) contain 1 to 30, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more nucleic acid bases. As described in this disclosure, in some embodiments, each nucleic acid base independently comprises an optionally substituted monocyclic, bicyclic, or polycyclic ring having at least one nitrogen ring atom; in some embodiments, each nucleic acid base independently comprises an optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, the wings, e.g., the first wing, the second wing, the 5'-wing, the 3'-wing, etc., are about 1 to 10 nucleic acid base lengths, e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10. In some embodiments, the wing is 1 nucleic acid base length. In some embodiments, the wing is about 2 nucleic acid base lengths. In some embodiments, the wing is about 3 nucleic acid base lengths. In some embodiments, the wing is about 4 nucleic acid base lengths. In some embodiments, the wing is about 5 nucleic acid base lengths. In some embodiments, the wing is about 6 nucleic acid base lengths. In some embodiments, the wing is about 7 nucleic acid base lengths. In some embodiments, each wing of the wing-core-wing structure independently has the lengths described herein. In some embodiments, two wings are the same length. In some embodiments, two wings are different lengths. In some embodiments, the core is about 5 to 25 nucleic acid base lengths, for example, about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleic acid base lengths. In some embodiments, the core is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or longer. In some embodiments, the core is about 5 nucleic acid base lengths. In some embodiments, the core is about 6 nucleic acid base lengths. In some embodiments, the core is about 7 nucleic acid base lengths. In some embodiments, the core is about 8 nucleic acid base lengths.In some embodiments, the core is approximately 9 nucleic acid bases long. In some embodiments, the core is approximately 10 nucleic acid bases long. In some embodiments, the core is approximately 11 nucleic acid bases long. In some embodiments, the core is about 12 nucleic acid bases long. In some embodiments, the core is about 13 nucleic acid bases long. In some embodiments, the core is about 14 nucleic acid bases long. In some embodiments, the core is about 15 nucleic acid bases long. For example, in some embodiments, the wing-core-wing is 2-9-6, 3-9-3, 3-9-4, 3-9-5, 4-7-4, 4-9-4, 4-9-5, 4-10-5, 4-11-4, 4-11-5, 5-7-5, 5-8-6, 5-9-3, 5-9-5, 5-10-4, 5-10-5, 6-7-6, 6-8-5, or 6-9-2. In some embodiments, it is 5-10-5.

[0170] In some embodiments, the wings include one or more glycosylation modifications. In some embodiments, the two wings of a wing-core-wing structure include different glycosylation patterns or different glycosylation modifications (and the oligonucleotides have or include an "asymmetrical" format). In some embodiments, the glycosylation provides improved stability and / or annealing properties compared to the absence of glycosylation.

[0171] In some embodiments, the first wing (e.g., the 5'-wing) has one or more 2'-OR modifications (wherein R is optionally replaced by C). 1~4 It includes (which is aliphatic). In some embodiments, each sugar in the first wing contains a 2'-OR modification. In some embodiments, the 2'-OR is a 2'-MOE. In some embodiments, each sugar in the first wing contains a 2'-MOE.

[0172] In some embodiments, the second wing (e.g., the 3'-wing) is one or more 2'-OR modifications (wherein R is optionally replaced by C). 1~4It includes (being aliphatic). In some embodiments, each sugar of the second wing contains a 2'-OR modification. In some embodiments, the 2'-OR is 2'-OMe. In some embodiments, each sugar of the second wing contains 2'-OMe. In some embodiments, the second wing, e.g., the 3'-wing, does not share the same sugar modification pattern as the first wing, e.g., the 5'-wing. In some embodiments, the second wing, e.g., the 3'-wing, does not contain the sugar modifications of the first wing, e.g., the 5'-wing. As those skilled in the art will understand, in some embodiments, the first wing may be the 3'-wing and the second wing may be the 5'-wing.

[0173] In some embodiments, the core comprises sugars 1 to 25, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or sugars that do not contain a 2'-OR group, or sugars that are not bicyclic or polycyclic sugars. In some embodiments, the core comprises sugars 1 to 25, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or sugars that do not contain a 2'-OR group. In some embodiments, the core comprises sugars 1 to 25, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or two 2'-H groups. In many embodiments, the core does not contain a 2'-OR group. In many embodiments, the sugars in the core region have two 2'-H groups.

[0174] In some embodiments, a specific sugar modification, e.g., 2'-MOE, provides higher stability under certain conditions compared to other sugar modifications, e.g., 2'-OMe. In some embodiments, the wing includes a 2'-MOE modification. In some embodiments, each nucleoside unit of the wing containing a pyrimidine base (e.g., C, U, T, etc.) includes a 2'-MOE modification. In some embodiments, each sugar unit of the wing includes a 2'-MOE modification. In some embodiments, each nucleoside unit of the wing containing a purine base (e.g., A, G, etc.) The nucleoside units are free from 2'-MOE modifications (for example, each such nucleoside unit contains 2'-OMe or does not contain 2'-modification). In some embodiments, each nucleoside unit of the purine base-containing wing contains a 2'-OMe modification. In some embodiments, each internucleotide bond at the 3' position of the sugar unit containing a 2'-MOE modification is a native phosphate bond.

[0175] In some embodiments, the wing does not contain a 2'-MOE modification. In some embodiments, the wing contains a 2'-OMe modification. In some embodiments, each nucleoside unit of the wing independently contains a 2'-OMe modification.

[0176] In some embodiments, the wings contain a bicyclic sugar. In some embodiments, each wing independently contains one or more bicyclic sugars.

[0177] In some embodiments, the bicyclic sugar is LNA, cEt, or BNA sugar.

[0178] In some embodiments, the MAPT oligonucleotide has a wing-core-wing structure. In some embodiments, the core contains one or more native DNA sugars. In some embodiments, the core contains five or more consecutive native DNA sugars. In some embodiments, the core contains 5-10, 5-15, 5-20, 5-25, 5-30, or 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more consecutive native DNA sugars. In some embodiments, the core contains 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more consecutive native DNA sugars. In some embodiments, the core contains 10 or more consecutive native DNA sugars. In some embodiments, the core can hybridize to a target mRNA, thereby forming a double-stranded structure that can be recognized by RNase H, allowing RNase H to cleave this mRNA.

[0179] In some embodiments, MAPT oligonucleotides have a wing-core-wing structure and an asymmetric format.

[0180] In some embodiments, in oligonucleotides having an asymmetric format, one wing differs from the other in terms of glycosylation or its pattern, or in terms of skeletal internucleotide linkage or its pattern, or in terms of skeletal chiral centers or its pattern. In some embodiments, MAPT oligonucleotides have an asymmetric format in that one wing contains a different glycosylation than the other wing. In some embodiments, MAPT oligonucleotides have an asymmetric format in that one wing contains a different glycosylation pattern than the other wing.

[0181] In some embodiments, MAPT oligonucleotides (or wings, cores, regions, blocks or any part thereof) are U.S. Patent Nos. 9,394,333, 9,744,183, 9,605,019, 9,598,458, 9,982,257, 1,016,0969, 1,047,999, 2020 / 0056173, 2018 / 0216107, 2019 / 0127733, and 1,045,0568. , U.S. Patent Application Publication No. 2019 / 0077817, U.S. Patent Application Publication No. 2019 / 0249173, U.S. Patent Application Publication No. 2019 / 0375774, International Publication No. 2018 / 223056, International Publication No. 2018 / 223073, International Publication No. 2018 / 223081, International Publication No. 2018 / 237194, International Publication No. 2019 / 032607, International Publication No. 2019 / 055951, International Publication No. 2019 / 075357, International Publication No. 2019 / 200185, International Publication No. 2019 / 217784, International Publication No. 201 Modifications, modification patterns, internucleotide links, internucleotide link patterns, chiral center patterns, wings, cores, blocks, regions, and / or formats (including, but not limited to, asymmetric formats) as described in 9 / 032612 and / or International Publication No. 2020 / 191252 (each of which describes modifications, modification patterns, internucleotide links, internucleotide link patterns, chiral center patterns, wings, cores, blocks, regions, and formats are incorporated herein by reference independently).

[0182] In some embodiments, the structure of the MAPT oligonucleotide is a wing-core structure or includes one.

[0183] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each independently comprise at least one 2'-MOE.

[0184] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each independently comprise at least one 2'-OMe.

[0185] In some embodiments, the structure of the MAPT oligonucleotide includes or consists of an asymmetric format. In some embodiments, the structure of the MAPT oligonucleotide includes or consists of a symmetric format.

[0186] In some embodiments, the structure of the MAPT oligonucleotide is or includes an asymmetric format, wherein the oligonucleotide structure is a wing-core-wing structure, where the format of the first wing differs from the format of the second wing. In some embodiments, the structure of the MAPT oligonucleotide is or includes an asymmetric format, wherein the oligonucleotide structure is a wing-core-wing structure, where the first and second wings differ in sugar modification (or combination or pattern thereof) and / or internucleotide bonding (or combination or pattern thereof). In some embodiments, the structure of the MAPT oligonucleotide is or includes an asymmetric format, wherein the oligonucleotide structure is a wing-core-wing structure, where the first and second wings differ in sugar modification (or combination or pattern thereof).

[0187] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where one wing comprises one type of sugar and the other comprises that type and a second type.

[0188] In some embodiments, the structure of a MAPT oligonucleotide comprises a core and at least one wing, the core comprising 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more consecutive 2'-deoxyribose sugars.

[0189] In some embodiments, the structure of a MAPT oligonucleotide comprises a core and at least one wing, the core comprising at least five consecutive 2'-deoxyribose sugars.

[0190] In some embodiments, the structure of a MAPT oligonucleotide comprises a core and at least one wing, the core comprising at least six consecutive 2'-deoxyribose sugars include.

[0191] In some embodiments, the structure of a MAPT oligonucleotide comprises a core and at least one wing, the core comprising at least seven consecutive 2'-deoxyribose sugars.

[0192] In some embodiments, the structure of a MAPT oligonucleotide comprises a core and at least one wing, where the core comprises at least eight consecutive 2'-deoxyribose sugars.

[0193] In some embodiments, the structure of a MAPT oligonucleotide comprises a core and at least one wing, wherein the core comprises at least nine consecutive 2'-deoxyribose sugars.

[0194] In some embodiments, the structure of a MAPT oligonucleotide comprises a core and at least one wing, where the core comprises at least 10 consecutive 2'-deoxyribose sugars.

[0195] In some embodiments, the MAPT oligonucleotide comprises at least three different types of internucleotide bonds.

[0196] In some embodiments, the MAPT oligonucleotide comprises at least one native phosphate internucleotide bond; at least one phosphorothioate; and at least one neutral or non-negatively charged internucleotide bond.

[0197] In some embodiments, the MAPT oligonucleotide comprises at least one native phosphate internucleotide bond; at least one chiral-controlled phosphorothioate; and at least one neutral or non-negatively charged internucleotide bond.

[0198] In some embodiments, the MAPT oligonucleotide comprises at least one native phosphate internucleotide bond; at least one phosphorothioate; and at least one chiral-controlled neutral or non-negatively charged internucleotide bond.

[0199] In some embodiments, the MAPT oligonucleotide comprises at least one native phosphate internucleotide bond; at least one chiral-controlled phosphorothioate; and at least one chiral-controlled neutral or non-negatively charged internucleotide bond.

[0200] In some embodiments, the structure of a MAPT oligonucleotide comprises a core and at least one wing, where the core comprises at least 12 consecutive 2'-deoxyribose sugars.

[0201] In some embodiments, the structure of a MAPT oligonucleotide comprises a core and at least one wing, where the core comprises at least 14 consecutive 2'-deoxyribose sugars.

[0202] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each comprise at least two different types of sugars.

[0203] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each comprise a 2'-DNA sugar (natural 2'-deoxyribose) and a 2'-modified sugar.

[0204] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each comprise a 2'-DNA sugar (natural 2'-deoxyribose) and a 2'-OMe sugar.

[0205] In some embodiments, the MAPT oligonucleotide comprises at least one natural 2'-deoxyribose sugar (unmodified DNA sugar), at least one LNA sugar, and at least one 2'-MOE sugar.

[0206] In some embodiments, the structure of the MAPT oligonucleotide is a wing-core-wing structure or comprises the same, where the first and second wings each comprise a natural 2'-deoxyribose (unmodified DNA sugar), an LNA sugar, and a 2'-MOE sugar.

[0207] In some embodiments, the MAPT oligonucleotide comprises at least one natural 2'-deoxyribose (unmodified DNA sugar), at least one LNA sugar, and at least one 2'-OMe sugar.

[0208] In some embodiments, the structure of the MAPT oligonucleotide is a wing-core-wing structure or comprises the same, where the first and second wings each comprise a natural 2'-deoxyribose (unmodified DNA sugar), an LNA sugar, and a 2'-OMe sugar.

[0209] In some embodiments, the structure of the MAPT oligonucleotide is a wing-core-wing structure or comprises the same, where the first and second wings each comprise at least three different types of sugars (e.g., selected from unmodified sugars and modified sugars having various modifications).

[0210] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where at least one wing comprises one or more (1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) LNA sugars.

[0211] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each comprise one or more 2'-MOE sugars and one or more LNA sugars.

[0212] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each comprise one or more LNA sugars.

[0213] In some embodiments, the structure of the MAPT oligonucleotide is a wing-core-wing structure or comprises one wing comprising one or more LNA sugars and one or more 2'-MOE sugars, and the other wing comprising one or more LNA sugars and one or more 2'-OMe sugars.

[0214] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where one wing comprises natural 2'-deoxyribose (unmodified DNA sugar), LNA sugar, and 2'-MOE sugar.

[0215] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where one wing comprises at least three different types of sugars.

[0216] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each comprise a natural 2'-deoxyribose (unmodified DNA sugar) and at least one modified sugar (compared to 2'-deoxyribose (unmodified DNA sugar)).

[0217] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each comprise native 2'-deoxyribose (unmodified DNA sugar) and at least two sugar modifications.

[0218] In some embodiments, the structure of the MAPT oligonucleotide is a wing, where the wing comprises at least three different types of sugars.

[0219] In some embodiments, the structure of the MAPT oligonucleotide includes a wing, where the wing includes at least one base.

[0220] In some embodiments, the structure of the MAPT oligonucleotide includes a wing, where the wing includes at least two bases.

[0221] In some embodiments, the structure of the MAPT oligonucleotide includes a wing, where the wing includes at least three bases.

[0222] In some embodiments, the structure of the MAPT oligonucleotide includes a wing, where the wing includes at least four bases.

[0223] In some embodiments, the structure of MAPT oligonucleotides includes wings, where the wings consist of 5 bases.

[0224] In some embodiments, the structure of the MAPT oligonucleotide includes a wing, where the wing contains at least 6 bases.

[0225] In some embodiments, the structure of the MAPT oligonucleotide includes a wing, where the wing contains at least seven bases.

[0226] In some embodiments, the structure of MAPT oligonucleotides includes wings, where the wings consist of eight bases.

[0227] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where the first and second wings each contain two different types of sugars.

[0228] In some embodiments, the structure of the MAPT oligonucleotide is a wing-core-wing structure or comprises such a structure, where one wing comprises at least one 2'-MOE sugar and the other wing comprises at least one 2'-OMe sugar.

[0229] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where one wing is a natural 2'-deoxyribonucleotide. It contains a sugar (unmodified DNA sugar) and at least one modified sugar.

[0230] In some embodiments, the structure of the MAPT oligonucleotide is or comprises a wing-core-wing structure, where one wing comprises natural 2'-deoxyribose (unmodified DNA sugar) and at least two modified sugars.

[0231] In some embodiments, the MAPT oligonucleotide may include any first wing, core and / or second wing as described herein or known in the Art.

[0232] In some embodiments, an oligonucleotide having a base sequence that is, contains, or includes a MAPT oligonucleotide sequence disclosed herein, or a base sequence that includes a span thereof, may include a first wing, a core, and / or a second wing as described herein or known in the Art.

[0233] Internucleotide bonding In some embodiments, oligonucleotides include base modifications, sugar modifications, and / or internucleotide linkage modifications. In this disclosure, various internucleotide links may be used to link units containing nucleic acid bases, such as nucleosides. In some embodiments, MAPT oligonucleotides include both one or more modified internucleotide links and one or more native phosphate links. As is widely known to those skilled in the art, native phosphate links are widely found in native DNA and RNA molecules; they have the structure -OP(O)(OH)O-, link sugars in DNA and RNA nucleosides, and can be in various salt forms at physiological pH (about 7.4), and native phosphate links are mainly -OP(O)(OH)O- - It exists in the form of a salt with the anion -OP(O)(SH)O-. A modified internucleotide bond, or unnatural phosphate bond, is an internucleotide bond that is not a natural phosphate bond or its salt form. Modified internucleotide bonds can also be in the form of a salt, depending on their structure. For example, as those skilled in the art will understand, a phosphorothioate internucleotide bond having the structure -OP(O)(SH)O- exists, for example, at physiological pH (about 7.4), as -OP(O)(SH)O-. - It can take on various salt forms with the O- anion.

[0234] In some embodiments, the oligonucleotide includes an internucleotide bond which is a modified internucleotide bond, such as a phosphorothioate, phosphorodithioate, methylphosphonic acid, phosphoramidic acid, thiophosphate, 3'-thiophosphate, or 5'-thiophosphate.

[0235] In some embodiments, the modified internucleotide bond is a chiral internucleotide bond containing chiral-bound phosphorus. In some embodiments, the chiral internucleotide bond is a phosphorothioate bond. In some embodiments, the chiral internucleotide bond is a non-negatively charged internucleotide bond. In some embodiments, the chiral internucleotide bond is a neutral internucleotide bond. In some embodiments, the chiral internucleotide bond is chiral-controlled with respect to its chiral-bound phosphorus. In some embodiments, the chiral internucleotide bond is stereochemically pure with respect to its chiral-bound phosphorus. In some embodiments, the chiral internucleotide bond is not chiral-controlled. In some embodiments, the pattern of the skeletal chiral centers includes or consists of the positions and bound phosphorus configurations (Rp or Sp) of the chiral-controlled internucleotide bond and the positions of the achiral internucleotide bond (e.g., the native phosphate bond).

[0236] In some embodiments, the oligonucleotide is U.S. Patent No. 9,394,333, U.S. Japanese Patent No. 9744183, U.S. Patent No. 9605019, U.S. Patent No. 9598458, U.S. Patent No. 9982257, U.S. Patent No. 10160969, U.S. Patent No. 10479995, U.S. Patent Application Publication No. 2020 / 0056173, U.S. Patent Application Publication No. 2018 / 0216107, U.S. Patent Application Publication No. 2019 / 0127733, U.S. Patent No. 10450568, U.S. Patent Application Publication No. 2019 / 0077817, U.S. National Patent Application Publication No. 2019 / 0249173, US Patent Application Publication No. 2019 / 0375774, International Publication No. 2018 / 223056, International Publication No. 2018 / 223073, International Publication No. 2018 / 223081, International Publication No. 2018 / 237194, International Publication No. 2019 / 032607, International Publication No. 2019 / 055951, International Publication No. 2019 / 075357, International Publication No. 2019 / 200185, International Publication International Publication No. 2019 / 217784, International Publication No. 2019 / 032612, and / or International Publication No. 2020 / 191252 (each of these internucleotide links (e.g., formula I, formula Ia, formula Ib, or formula Ic, formula In-1, formula In-2, formula In-3, formula In-4, formula II, formula II-a-1, formula II-a-2, formula II-b-1, formula II-b-2, formula II-c-1, formula II-c-2, formula II-d-1, formula II-d-2, etc.) The modified internucleotide bond includes, independently, a modified internucleotide bond as described herein (independently, as incorporated herein by reference) (for example, a modified internucleotide bond having the structure of formula I, formula Ia, formula Ib, or formula Ic, formula In-1, formula In-2, formula In-3, formula In-4, formula II, formula II-a-1, formula II-a-2, formula II-b-1, formula II-b-2, formula II-c-1, formula II-c-2, formula II-d-1, formula II-d-2, etc., or a salt form thereof). In some embodiments, the modified internucleotide bond is a non-negatively charged internucleotide bond. In some embodiments, the oligonucleotide provided includes one or more non-negatively charged internucleotide bonds. In some embodiments, the non-negatively charged internucleotide bond is a positively charged internucleotide bond. In some embodiments, the non-negatively charged internucleotide bond is a neutral internucleotide bond.In some embodiments, the Disclosure provides oligonucleotides comprising one or more neutral internucleotide bonds. In some embodiments, non-negatively charged internucleotide bonds or neutral internucleotide bonds (e.g., formulas In-1, In-2, In-3, In-4, formula II, formula II-a-1, formula II-a-2, formula II-b-1, formula II-b-2, formula II-c-1, formula II-c-2, formula II-d-1, formula II-d-2, etc.) are U.S. Patent Nos. 9,394,333, 9,744,183, 9,605,019, 9,598,458, 9,982,257, 1,016,0969, 1,047,999,5, U.S. Patent Publication No. 2020 / 0056173, U.S. Patent Publication No. 2018 / 0216107, U.S. Patent Publication No. 2019 / Patent No. 0127733, U.S. Patent No. 10450568, U.S. Patent Application Publication No. 2019 / 0077817, U.S. Patent Application Publication No. 2019 / 0249173, U.S. Patent Application Publication No. 2019 / 0375774, International Publication No. 2018 / 223056, International Publication No. 2018 / 223073, International Publication No. 2018 / 223081, International Publication No. 20 As described in International Publication No. 18 / 237194, International Publication No. 2019 / 032607, International Publication No. 2019 / 055951, International Publication No. 2019 / 075357, International Publication No. 2019 / 200185, International Publication No. 2019 / 217784, International Publication No. 2019 / 032612, and / or International Publication No. 2020 / 191252.In some embodiments, non-negatively charged internucleotide bonds or neutral internucleotide bonds are described in International Publication Nos. 2018 / 223056, 2019 / 032607, 2019 / 075357, 2019 / 032607, 2019 / 075357, 2019 / 200185, and 2019 / 217784. Formulas In-1, In-2, In-3, In-4, II, II-a-1, II-a-2, II-b-1, II-b-2, II-c-1, II-c, as described in International Publication No. 2019 / 032612 and / or International Publication No. 2020 / 191252 (each of these internucleotide links is independently incorporated herein by reference). These include equations -2, equation II-d-1, equation II-d-2, etc.

[0237] In some embodiments, non-negatively charged internucleotide bonds can enhance the delivery and / or activity of oligonucleotides (e.g., the ability to reduce the level, activity, and / or expression of a target gene or its gene product).

[0238] In some embodiments, the modified internucleotide bond (e.g., a non-negatively charged internucleotide bond) comprises an optionally substituted triazolyl. In some embodiments, the modified internucleotide bond (e.g., a non-negatively charged internucleotide bond) comprises an optionally substituted alkynyl. In some embodiments, the modified internucleotide bond comprises a triazole or alkyne moiety. In some embodiments, the triazole moiety, e.g., a triazolyl group, is optionally substituted. In some embodiments, the triazole moiety, e.g., a triazolyl group, is substituted. In some embodiments, the triazole moiety is unsubstituted. In some embodiments, the modified internucleotide bond comprises an optionally substituted cyclic guanidine moiety. In some embodiments, the modified internucleotide bond comprises an optionally substituted cyclic guanidine moiety, [ka] It has the structure (wherein W is O or S). In some embodiments, W is O. In some embodiments, W is S. In some embodiments, the non-negative charge internucleotide bond is stereochemically controlled.

[0239] In some embodiments, the non-negatively charged internucleotide bond or neutral internucleotide bond is an internucleotide bond containing a triazole moiety. In some embodiments, the non-negatively charged internucleotide bond or non-negatively charged internucleotide bond contains an optionally substituted triazolyl group. In some embodiments, the internucleotide bond containing a triazole moiety (e.g., an optionally substituted triazolyl group) [ka] It has the structure. In some embodiments, the internucleotide bond containing the triazole moiety is [ka] It has the following structure. In some embodiments, the internucleotide bond, for example, the non-negatively charged internucleotide bond, the neutral internucleotide bond, includes a cyclic guanidine moiety. In some embodiments, the internucleotide bond including the cyclic guanidine moiety is [ka] It has the following structure. In some embodiments, the non-negatively charged internucleotide bond or the neutral internucleotide bond is [ka] The structure is selected from (wherein W is O or S) or includes such a structure.

[0240] In some embodiments, internucleotide binding is performed by Tmg groups [ka] Includes. In some embodiments, the internucleotide bond includes a Tmg group. [ka] It has the structure of a ("Tmg internucleotide bond"). In some embodiments, the neutral internucleotide bond includes internucleotide bonds of PNA and PMO and a Tmg internucleotide bond.

[0241] In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 3-20 membered heterocyclyl or heteroaryl group having 1-10 heteroatoms. In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 3-20 membered heterocyclyl or heteroaryl group having 1-10 heteroatoms such that at least one heteroatom is nitrogen. In some embodiments, such heterocyclyl or heteroaryl group is a 5-membered ring. In some embodiments, such heterocyclyl or heteroaryl group is a 6-membered ring.

[0242] In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 5-20 membered ring heteroaryl group having 1-10 heteroatoms. In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 5-20 membered ring heteroaryl group having 1-10 heteroatoms such that at least one heteroatom is nitrogen. In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 5-6 membered ring heteroaryl group having 1-4 heteroatoms such that at least one heteroatom is nitrogen. In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 5 membered ring heteroaryl group having 1-4 heteroatoms such that at least one heteroatom is nitrogen. In some embodiments, the heteroaryl group is directly bonded to the phosphate group. In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted triazolyl group. In some embodiments, the non-negatively charged internucleotide bond comprises an unsubstituted triazolyl group, for example, [ka] Includes. In some embodiments, the non-negatively charged internucleotide bond is a substituted triazolyl group, for example, [ka] Includes.

[0243] In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 5-20 member heterocyclyl group having 1-10 heteroatoms. In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 5-20 member heterocyclyl group having 1-10 heteroatoms, where at least one heteroatom is nitrogen. In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 5-6 member heterocyclyl group having 1-4 heteroatoms, where at least one heteroatom is nitrogen. In some embodiments, the non-negatively charged internucleotide bond comprises an optionally substituted 5 member heterocyclyl group having 1-4 heteroatoms, where at least one heteroatom is nitrogen. In some embodiments, at least two heteroatoms are nitrogen. In some embodiments, the heterocyclyl group is directly bonded to the conjugated phosphorus. In some embodiments, the heterocyclyl group is bound to the bound phosphorus via a linker, for example, when the heterocyclyl group is part of a guanidine moiety that is directly bound to the bound phosphorus at its =N-. In some embodiments, the non-negatively charged internucleotide bond is optionally substituted. [ka] It contains a group. In some embodiments, the non-negatively charged internucleotide bond is substituted. There are [ka] It contains a group. In some embodiments, the non-negatively charged internucleotide bond is [ka] Includes a group. In some embodiments, each R 1 These are C which are replaced independently and by choice. 1~6 It is alkyl. In some embodiments, each R 1These are independently methyl.

[0244] In some embodiments, modified internucleotide bonds, such as non-negatively charged internucleotide bonds, each comprises a triazole or alkyne moiety that is optionally substituted. In some embodiments, the modified internucleotide bond comprises a triazole moiety. In some embodiments, the modified internucleotide bond comprises an unsubstituted triazole moiety. In some embodiments, the modified internucleotide bond comprises a substituted triazole moiety. In some embodiments, the modified internucleotide bond comprises an alkyl moiety. In some embodiments, the modified internucleotide bond comprises an optionally substituted alkynyl group. In some embodiments, the modified internucleotide bond comprises an unsubstituted alkynyl group. In some embodiments, the modified internucleotide bond comprises a substituted alkynyl group. In some embodiments, the alkynyl group is directly bonded to the phosphate group.

[0245] In some embodiments, the oligonucleotide comprises different types of internucleotide phosphate bonds. In some embodiments, the chiral-controlled oligonucleotide comprises at least one native phosphate bond and at least one modified (unnatural) internucleotide bond. In some embodiments, the oligonucleotide comprises at least one native phosphate bond and at least one phosphorothioate. In some embodiments, the oligonucleotide comprises at least one non-negatively charged internucleotide bond. In some embodiments, the oligonucleotide comprises at least one native phosphate bond and at least one non-negatively charged internucleotide bond.

[0246] While not wishing to be constrained by any particular theory, this disclosure notes that neutral internucleotide bonds may be more hydrophobic than phosphorothioate internucleotide bonds (PS), and PS may be more hydrophobic than native phosphate bonds (PO). Typically, unlike PS or PO, neutral internucleotide bonds have a lower supported charge. While not wishing to be constrained by any particular theory, this disclosure notes that incorporating one or more neutral internucleotide bonds into an oligonucleotide may increase the oligonucleotide's ability to be taken up by cells and / or escape from endosomes. While not wishing to be constrained by any particular theory, this disclosure notes that the incorporation of one or more neutral internucleotide bonds may be used to modulate the melting temperature of the double helix formed between the oligonucleotide and its target nucleic acid.

[0247] While not intending to be bound by any particular theory, this disclosure notes that incorporating one or more non-negatively charged internucleotide bonds, such as neutral internucleotide bonds, into an oligonucleotide may increase the oligonucleotide's ability to mediate functions such as gene knockdown. In some embodiments, an oligonucleotide, such as a MAPT oligonucleotide having the ability to mediate knockdown at a level of nucleic acid or the product encoded therein, comprises one or more non-negatively charged internucleotide bonds.

[0248] As has been well demonstrated in many embodiments, the oligonucleotides of this disclosure comprise two or more distinct internucleotide bonds. In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide bond and a non-negatively charged internucleotide bond. In some embodiments, the oligonucleotide comprises a phosphorothioate internucleotide bond, a non-negatively charged internucleotide bond, and a native phosphate bond. In some embodiments, the non-negatively charged internucleotide bond is a neutral internucleotide bond. In some embodiments, the non-negatively charged internucleotide bond is n001. In some embodiments, each phosphorothioate internucleotide bond is independently chiralized. In some embodiments, each chiralized internucleotide bond is independently chiralized.

[0249] Typical bonds, such as those found in natural DNA and RNA, involve an internucleotide bond forming a bond with two sugars (which may be unmodified or modified as described herein). In many embodiments, as illustrated herein, the internucleotide bond forms a bond at its 5' carbon to one optionally modified ribose or deoxyribose with its oxygen atom or heteroatom, and a bond at its 3' carbon to the other optionally modified ribose or deoxyribose. In some embodiments, each nucleoside unit linked by the internucleotide bond independently contains an optionally substituted nucleic acid base of A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U.

[0250] In some embodiments, MAPT oligonucleotides include internucleotide bonds in which the negatively charged, non-crosslinked oxygen of a canonical phosphate diester bond is replaced by an uncharged alkyl substituent such as a methyl (Met) or ethyl (Et) group, as is the case with p-alkylphosphonic acid nucleic acids (phNA), such as p-methyl or p-ethyl phNA. See, for example, Micklefield et al. 2001 Curr. Med. Chem. 8, 1157-1179; and Arangundy-Franklin et al. 2019 Nat. Chem. 11, 533-542.

[0251] In some embodiments, the MAPT oligonucleotide is a phosphonomethyl-threosyl nucleic acid (tPhoNA) and / or contains a phosphonomethyl-threosyl internucleotide bond. Liu et al. 2018 J. Am. Chem. Soc. 140, 6690-6699.

[0252] As those skilled in the art will understand, this disclosure does not cover many other types of internucleotide linkages, e.g., U.S. Patent Nos. 3,687,808; 4,469,863; 4,476,301; 5,177,195; 5,023,243; 5,034,506; 5,166,315; 5,185,444; 5,188,897; 5,214,134; 5,216,141; 5,235,033; 5,264,423; 5,264,564; 5,276,01 No. 9; No. 5,278,302; No. 5,286,717; No. 5,321,131; No. 5,399,676; No. 5,405,938; No. 5,405,939; No. 5,434,257; Same No. 5,453,496; Same No. 5,455,233; Same No. 5,466,677; Same No. 5,466,677; Same No. 5,470,967; Same No. 5,476,925; Same No. 5,489,677; Same No. 5, 519,126; 5,536,821; 5,541,307; 5,541,316; 5,550,111; 5,561,225; 5,563,253; 5,571 ,799; 5,587,361; 5,596,086; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070 No. 5,625,050; No. 5,633,360; No. 5,64,562; No. 5,663,312; No. 5,677,437; No. 5,677,439; No. 6,160,109; 6,239,265; 6,028,188; 6,124,445; 6,169,170; 6,172,209; 6,277,603; 6,326,199; 6,34 You may use the documents listed in No. 6,614; No. 6,444,423; No. 6,531,590; No. 6,534,639; No. 6,608,035; No. 6,683,167; No. 6,858,715; No. 6,867,294; No. 6,878,805; No. 7,015,315; No. 7,041,816; No. 7,273,933; No. 7,321,029; or No. RE39464.In some embodiments, modified internucleotide bondage is used in U.S. Patent Publications 9394333, 9744183, 9605019, 9598458, 9982257, U.S. Patent Application Publication 10160969, U.S. Patent Application Publication 10479995, U.S. Patent Application Publication 2020 / 0056173, U.S. Patent Application Publication 2018 / 0216107, U.S. Patent Application Publication 2019 / 0127733, U.S. Patent Application Publication 10450568, U.S. Patent Application Publication 2019 / 0077817, U.S. Patent Application Publication 2019 / 0249173, U.S. Patent Application Publication 2019 / 0375774, and International Publication. These are described in International Publication No. 2018 / 223056, International Publication No. 2018 / 223073, International Publication No. 2018 / 223081, International Publication No. 2018 / 237194, International Publication No. 2019 / 032607, International Publication No. 2019 / 055951, International Publication No. 2019 / 075357, International Publication No. 2019 / 200185, International Publication No. 2019 / 217784, International Publication No. 2019 / 032612 and / or International Publication No. 2020 / 191252 (each of these nucleic acid bases, sugars, internucleotide bonds, chiral auxiliary groups / reagents, and oligonucleotide synthesis techniques (reagents, conditions, cycles, etc.) are independently incorporated herein by reference).

[0253] In some embodiments, the oligonucleotide comprises one or more nucleotides independently containing phosphorus modifications that tend to “auto-release” under certain conditions. That is, under certain conditions, a particular phosphorus modification is designed to autocleave from the oligonucleotide to provide, for example, a native phosphate bond. For specific examples of such phosphorus modification groups, see U.S. Patent No. 9,982,257. In some embodiments, the auto-releasing group includes a morpholino group. In some embodiments, the auto-releasing group is characterized by its ability to deliver a drug to an internucleotide phosphate linker, which promotes further modification of the phosphorus atom, such as desulfurization. In some embodiments, the drug is water, and the further modification is the formation of a native phosphate bond by hydrolysis.

[0254] In some embodiments, the oligonucleotide comprises one or more internucleotide bonds that enhance one or more of the pharmaceutically active properties and / or activity of the oligonucleotide. In the art, it is well known that certain oligonucleotides are readily degraded by nucleases and exhibit low cellular uptake across the cytoplasmic membrane (Poijarvi-Virta). et al., Curr. Med. Chem. (2006), 13(28);3441-65;Wagner et al., Med. Res. Rev. (2000), 20(6):417-51; Peyrottes et al., Mini Rev. Med. Chem. (2004), 4(4):395-408; Gosselin et al., (1996), 43(1):196-208; Bologna et al., (2002), Antisense & Nucleic Acid Drug Development 12:33-41). Vives et al. (Nucleic Acids Research (1999), 27(20):4071-76) reported that tert-butylSATE prooligonucleotides showed a significant increase in cell permeability compared to parent oligonucleotides under certain conditions.

[0255] Various types of internucleotide bonds can be used in combination with other structural elements, such as sugars, to achieve desired oligonucleotide properties and / or activity. For example, the Disclosure conventionally and optionally utilizes modified internucleotide bonds and modified sugars together with natural phosphate bonds and natural sugars when designing oligonucleotides. In some embodiments, the Disclosure provides oligonucleotides comprising one or more modified sugars. In some embodiments, the Disclosure provides oligonucleotides comprising one or more modified sugars and one or more modified internucleotide bonds (one or more of which are natural phosphate bonds).

[0256] Oligonucleotide composition In particular, this disclosure provides various oligonucleotide compositions. In some embodiments, this disclosure provides oligonucleotide compositions of the oligonucleotides described herein. In some embodiments, the oligonucleotide composition, for example, the oligonucleotide composition, comprises a plurality of oligonucleotides described herein. In some embodiments, the oligonucleotide composition, for example, the MAPT oligonucleotide composition, is chiral-controlled. In some embodiments, the oligonucleotide composition, for example, the MAPT oligonucleotide composition, is not chiral-controlled (sterically random).

[0257] The bound phosphorus in natural phosphate bonds is achiral. The bound phosphorus in many modified internucleotide bonds, such as phosphorothioate internucleotide bonds, is chiral. In some embodiments, during the preparation of oligonucleotide compositions (e.g., in conventional phosphoramidite oligonucleotide synthesis), the arrangement of chiral bound phosphorus is not intentionally designed or controlled, resulting in a chiralally uncontrolled (stereorandom) oligonucleotide composition (a substantially racemic formulation) which is a random complex mixture of various stereoisomers (diastereoisomers) - typically for oligonucleotides having n chiral internucleotide bonds (where the bound phosphorus is chiral), 2 n It becomes 2 stereoisomers (for example, when n is 10, 2 10 = 1,032; when n is 20, 2 20 (=1,048,576). These stereoisomers have the same chemical structure, but differ in the stereochemical pattern of their phosphorus bonds.

[0258] In some embodiments, sterically random oligonucleotide compositions possess properties and / or activities sufficient for specific purposes and / or applications. In some embodiments, sterically random oligonucleotide compositions may be less expensive, easier, and / or simpler to produce than chiral-controlled oligonucleotide compositions. However, because stereoisomers in sterically random compositions can have various properties, activities, and / or toxicity, inconsistent therapeutic effects and / or unintended side effects may occur with sterically random compositions, particularly when compared to certain chiral-controlled oligonucleotide compositions of the same chemical composition.

[0259] Chiral-controlled oligonucleotide composition In some embodiments, the disclosure includes techniques for designing and preparing chiral-controlled oligonucleotide compositions. The rheotide composition comprises a plurality of oligonucleotides at a controlled / predetermined (not random, as in the case of a sterically random composition) level, wherein these oligonucleotides share the same binding phosphorus stereochemistry at one or more chiral internucleotide bonds (chiral controlled internucleotide bonds). In some embodiments, the plurality of oligonucleotides share the same skeletal chiral center pattern (binding phosphorus stereochemistry). In some embodiments, the skeletal chiral center pattern is as described in this disclosure. In some embodiments, the plurality of oligonucleotides share a common chemical configuration. In some embodiments, they are structurally identical.

[0260] For example, in some embodiments, the present disclosure provides an oligonucleotide composition comprising a plurality of oligonucleotides, wherein one of the plurality of oligonucleotides is 1) Common base sequence, and 2) One or more (for example, approximately 1-50, 1-40, 1-30, 1-25, 1-20, 1-15, 1-10, 5-50, 5-40, 5-30, 5-25, 5-20, 5-15, 5-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 or more) chiral internucleotide bonds with independently identical bonding phosphorus stereochemistry ("chiral-controlled internucleotide bonds") share; Here, the levels of multiple oligonucleotides in the composition are not random (for example, they are controlled / predetermined as described herein).

[0261] In some embodiments, the present disclosure provides an oligonucleotide composition comprising a plurality of oligonucleotides, wherein the plurality of oligonucleotides are 1) Common base sequence, and 2) One or more (for example, approximately 1-50, 1-40, 1-30, 1-25, 1-20, 1-15, 1-10, 5-50, 5-40, 5-30, 5-25, 5-20, 5-15, 5-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 or more) chiral internucleotide bonds with independently identical bonding phosphorus stereochemistry ("chiral-controlled internucleotide bonds") share; The composition features higher purity for some of the oligonucleotides compared to a substantially racemic formulation of oligonucleotides sharing a common base sequence.

[0262] In some embodiments, the present disclosure provides an oligonucleotide composition comprising a plurality of oligonucleotides, wherein the plurality of oligonucleotides are 1) Common base sequence, and 2) One or more (for example, approximately 1-50, 1-40, 1-30, 1-25, 1-20, 1-15, 1-10, 5-50, 5-40, 5-30, 5-25, 5-20, 5-15, 5-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 or more) chiral internucleotide bonds with independently identical bonding phosphorus stereochemistry ("chiral-controlled internucleotide bonds") share; Here, approximately 1% to 100% of all oligonucleotides in a composition sharing a common base sequence (e.g., approximately 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or approximately 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%). , 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) are oligonucleotides among multiple others.

[0263] In some embodiments, the percentage / level of one of the oligonucleotides is (DS) nc is or at least (DS) ncIn the formula, DS is 90% to 100%, and nc is the number of chiral-controlled internucleotide bonds. In some embodiments, nc is 5, 6, 7, 8, 9, 10 or more. In some embodiments, the percentage / level is at least 10%. In some embodiments, the percentage / level is at least 20%. In some embodiments, the percentage / level is at least 30%. In some embodiments, the percentage / level is at least 40%. In some embodiments, the percentage / level is at least 50%. In some embodiments, the percentage / level is at least 60%. In some embodiments, the percentage / level is at least 65%. In some embodiments, the percentage / level is at least 70%. In some embodiments, the percentage / level is at least 75%. In some embodiments, the percentage / level is at least 80%. In some embodiments, the percentage / level is at least 85%. In some embodiments, the percentage / level is at least 90%. In some embodiments, the percentage / level is at least 95%.

[0264] In some embodiments, the oligonucleotides share a common skeletal bonding pattern. In some embodiments, each of the oligonucleotides independently has an internucleotide bond of a specific chemical configuration (e.g., -OP(O)(SH)-O-) or a salt form thereof (e.g., -OP(O)(SNa)-O-) at each internucleotide binding site. In some embodiments, the internucleotide bonds at each internucleotide binding site are of the same configuration. In some embodiments, the internucleotide bonds at each internucleotide binding site are of different configurations.

[0265] In some embodiments, several oligonucleotides share a common chemical structure. In some embodiments, several oligonucleotides have the same common chemical structure. In some embodiments, several oligonucleotides have two or more common chemical structures. In some embodiments, several oligonucleotides are independently a specific oligonucleotide or a pharmaceutically acceptable salt thereof, or an oligonucleotide having the same chemical structure as that specific oligonucleotide or a pharmaceutically acceptable salt thereof. In some embodiments, about 1% to 100% of all oligonucleotides in a composition sharing a common chemical structure (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 20%, 30%, 4%). 0%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) are oligonucleotides among several. In some embodiments, the percentage of the level is (DS) nc is or at least (DS) nc In the formula, DS is 90% to 100%, and nc is the number of chiral-controlled internucleotide bonds. In some embodiments, nc is 5, 6, 7, 8, 9, 10 or more. In some embodiments, the level is at least 10%. In some embodiments And the level is at least 20%. In some embodiments, the level is at least 30%. In some embodiments, the level is at least 40%. In some embodiments, the level is at least 50%. In some embodiments, the level is at least 60%. In some embodiments, the level is at least 65%. In some embodiments, the level is at least 70%. In some embodiments, the level is at least 75%. In some embodiments, the level is at least 80%. In some embodiments, the level is at least 85%. In some embodiments, the level is at least 90%. In some embodiments, the level is at least 95%.

[0266] In some embodiments, each phosphorothioate internucleotide bond is independently a chiralally controlled internucleotide bond.

[0267] In some embodiments, this disclosure a) Common base sequence; b) Common skeletal connection patterns; c) Common skeletal chiral center pattern We provide a chiral controlled oligonucleotide composition containing multiple oligonucleotides of a specific oligonucleotide type characterized by; The composition is purified for specific oligonucleotide types compared to substantially racemic formulations of oligonucleotides having the same common base sequence.

[0268] In some embodiments, this disclosure a) Common base sequence; b) Common skeletal connection patterns; c) Common skeletal chiral center pattern We provide a chiral controlled oligonucleotide composition containing multiple oligonucleotides of a specific oligonucleotide type characterized by; Here, each of the oligonucleotides contains at least one internucleotide bond containing a common binding phosphorus in an Sp configuration; The composition is purified for specific oligonucleotide types compared to substantially racemic formulations of oligonucleotides having the same common base sequence.

[0269] A common skeletal chiral center pattern, as those skilled in the art will understand, includes at least one Rp or at least one Sp. Specific skeletal chiral center patterns are exemplified, for example, in Table 1A.

[0270] In some embodiments, chiral-controlled oligonucleotide compositions are purified for specific oligonucleotide types compared to substantially racemic formulations of oligonucleotides that share the same common base sequence and common skeletal bonding pattern.

[0271] In some embodiments, among a plurality of oligonucleotides, for example, of a particular oligonucleotide type, have a common skeletal phosphorus modification pattern and a common nucleoside modification pattern. In some embodiments, among a plurality of oligonucleotides, have a common sugar modification pattern. In some embodiments, among a plurality of oligonucleotides, have a common base modification pattern. In some embodiments, among a plurality of oligonucleotides, have a common nucleoside modification pattern. In some embodiments, a plurality Our oligonucleotides have the same chemical composition. In many embodiments, the oligonucleotides among several are identical. In some embodiments, the oligonucleotides among several are the same oligonucleotide (as those skilled in the art will understand, such oligonucleotides may each exist independently as one of various forms of the oligonucleotide and may be the same or different forms of the oligonucleotide). In some embodiments, the oligonucleotides among several are each independently the same oligonucleotide or a pharmaceutically acceptable salt thereof.

[0272] In some embodiments, the disclosure provides chiral-controlled oligonucleotide compositions of many oligonucleotides, for example, those listed in Table 1, whose "stereochemistry / bonding" includes S and / or R. In some embodiments, each of the plurality of oligonucleotides is independently, optionally in various forms, of a specific oligonucleotide listed in Table 1, whose "stereochemistry / bonding" includes S and / or R. In some embodiments, each of the plurality of oligonucleotides is independently, of a specific oligonucleotide listed in Table 1, whose "stereochemistry / bonding" includes S and / or R, or a pharmaceutically acceptable salt thereof.

[0273] In some embodiments, the levels of multiple oligonucleotides in a composition can be determined as the product of the diastereopurities of each chiral-controlled internucleotide bond in the oligonucleotide. In some embodiments, the diastereopurity of an internucleotide bond linking two nucleosides in an oligonucleotide (or nucleic acid) is expressed by the diastereopurity of an internucleotide bond of a dimer linking the same two nucleosides, where this dimer is prepared under equivalent conditions, and in some examples, under the same synthetic cycle conditions.

[0274] In some embodiments, the chiral internucleotide bonds are independently and entirely chiral-controlled, resulting in a fully chiral-controlled oligonucleotide composition. In other embodiments, not all chiral internucleotide bonds are chiral-controlled, resulting in a partially chiral-controlled oligonucleotide composition.

[0275] Oligonucleotides may contain, or consist of, various skeletal chiral center patterns (stereochemical patterns of chiral-bound phosphorus). Certain useful skeletal chiral center patterns are described in this disclosure. In some embodiments, multiple oligonucleotides share a common skeletal chiral center pattern, which is or includes the patterns described in this disclosure (e.g., those in "Stereochemistry and Patterns of Skeletal Chiral Centers," the skeletal chiral center patterns of chiral-controlled oligonucleotides in Table 1, etc.).

[0276] In some embodiments, a chiral-controlled oligonucleotide composition is a chiralally pure (or sterically pure, stereochemically pure) oligonucleotide composition comprising a plurality of oligonucleotides, wherein these oligonucleotides are independently the same stereoisomer [including that each chiral element of the oligonucleotide, including each chiral-linked phosphorus, is independently defined (sterically defined)]. A chirally pure (or sterically pure, stereochemically pure) oligonucleotide composition of oligonucleotide stereoisomers is free from other stereoisomers (as those skilled in the art will understand, one or more unintended stereoisomers may be present as impurities, for example, from preparation, storage, etc.).

[0277] Chiral-controlled oligonucleotide compositions can demonstrate several advantages over sterically random oligonucleotide compositions. In particular, chiral-controlled oligonucleotides Gonucleotide compositions exhibit greater uniformity in terms of oligonucleotide structure compared to corresponding sterically random oligonucleotide compositions. By controlling stereochemistry, compositions of individual stereoisomers can be prepared and evaluated, thus enabling the development of chiral-controlled oligonucleotide compositions with desired properties and / or activities. In some embodiments, chiral-controlled oligonucleotide compositions offer, for example, better delivery, stability, clearance, activity, selectivity, and / or toxicity profiles compared to corresponding sterically random oligonucleotide compositions. In some embodiments, chiral-controlled oligonucleotide compositions offer better efficacy, fewer side effects, and / or more convenient and effective dosage regimens. In particular, by utilizing the skeletal chiral center patterns as described herein, controlled cleavage of oligonucleotide targets can be achieved (e.g., transcripts such as pre-mRNA and mature mRNA; including control of cleavage sites, rate and / or degree of cleavage at cleavage sites, and / or overall rate and degree of cleavage).

[0278] In some embodiments, the oligonucleotides in the provided composition, for example, the chiral-controlled oligonucleotide composition, are MAPT oligonucleotides as described herein.

[0279] In some embodiments, the Disclosure provides sterically random oligonucleotide compositions, such as sterically random MAPT oligonucleotide compositions. In some embodiments, the Disclosure provides sterically random MAPT oligonucleotide compositions having the ability to reduce the level, activity, or expression of a MAPT gene or its gene product. In some embodiments, the Disclosure provides sterically random MAPT oligonucleotide compositions having the ability to reduce the level, activity, or expression of a MAPT gene or its gene product, wherein the nucleotide sequence of the MAPT oligonucleotide is, or includes, the nucleotide sequences disclosed herein (e.g., the nucleotide sequences in Table 1, where each T can be independently replaced by a U, and vice versa), or includes its span (e.g., at least 10 or 15 adjacent nucleotides).

[0280] In some embodiments, the oligonucleotide composition comprises one or more sterically controlled (chiral controlled; in some embodiments, sterically pure) internucleotide bonds and one or more sterically random internucleotide bonds. In some embodiments, the MAPT oligonucleotide composition comprises one or more sterically controlled (chiral controlled; in some embodiments, sterically pure) internucleotide bonds and one or more sterically random internucleotide bonds.

[0281] In some embodiments, the oligonucleotide composition comprises one or more sterically controlled (e.g., chiral controlled or sterically pure) internucleotide bonds and one or more sterically random internucleotide bonds. Such oligonucleotides can target a variety of targets, have a variety of base sequences, and have the ability to function by one or more of a variety of modalities (e.g., RNase H mechanisms, steric hindrance, double-stranded or single-stranded RNA interference, exon skipping regulation, CRISPR, aptamers, etc.).

[0282] In some embodiments, the disclosure provides chiral-controlled oligonucleotide compositions, such as chiral-controlled MAPT oligonucleotide compositions. In some embodiments, the provided chiral-controlled oligonucleotide compositions comprise a plurality of oligonucleotides having the same chemical structure, such as MAPT oligonucleotides, and having one or more internucleotide bonds. In some embodiments, for example, the plurality of oligonucleotides in the chiral-controlled oligonucleotide composition are a plurality of oligonucleotides selected from Table 1. A nucleotide, where the oligonucleotide contains at least one Rp or Sp-linked phosphorus in a chiral-controlled internucleotide bond. In some embodiments, for example, in a chiral-controlled oligonucleotide composition, the plurality of oligonucleotides are plurality of oligonucleotides selected from Table 1, where each phosphorothioate internucleotide bond in the oligonucleotide is independently chiral-controlled (each phosphorothioate internucleotide bond is independently Rp or Sp). In some embodiments, the oligonucleotide composition, for example, a MAPT oligonucleotide composition, is a substantially pure formulation of a single oligonucleotide, so to speak, in that any oligonucleotides in the composition that are not that single oligonucleotide are impurities from the preparation process of the single oligonucleotide, possibly after certain purification steps. In some embodiments, the single oligonucleotide is an oligonucleotide from Table 1, where each chiral internucleotide bond in the oligonucleotide is chiral-controlled (e.g., indicated as S or R rather than X in "Stereochemistry / Bonding").

[0283] In some embodiments, chiral-controlled oligonucleotide compositions may exhibit increased activity and / or stability, increased delivery, and / or reduced ability to induce adverse effects such as complement and TLR9 activation compared to the corresponding sterically random oligonucleotide compositions. In some embodiments, sterically random (non-chiral-controlled) oligonucleotide compositions differ from chiral-controlled oligonucleotide compositions in that their corresponding oligonucleotides do not contain any chiral-controlled internucleotide bonds, but sterically random oligonucleotide compositions are otherwise identical to chiral-controlled oligonucleotide compositions.

[0284] In some embodiments, the disclosure relates to chiral-controlled MAPT oligonucleotide compositions having the ability to reduce the level, activity, or expression of MAPT genes or their gene products.

[0285] In some embodiments, the Disclosure provides chiral-controlled MAPT oligonucleotide compositions comprising a plurality of oligonucleotides that have the ability to reduce the level, activity, or expression of the MAPT gene or its gene product and that share a common nucleotide sequence that is or contains the nucleotide sequence disclosed herein (e.g., in Table 1, where each T can be independently replaced by U, and vice versa).

[0286] In some embodiments, the chiral-controlled oligonucleotide composition provided is a chiral-controlled MAPT oligonucleotide composition comprising multiple MAPT oligonucleotides. In some embodiments, the chiral-controlled oligonucleotide composition is a chiralally pure (or "stereochemically pure") oligonucleotide composition. In some embodiments, the disclosure provides chirally pure oligonucleotide compositions of the oligonucleotides listed in Table 1 (e.g., 1A), where each chiral internucleotide bond of this oligonucleotide is independently chiral-controlled (Rp or Sp, e.g., R or S instead of X in "stereochemistry / bond"). As those skilled in the art will understand, achieving complete (absolute 100%) chemical selectivity is rare, if not impossible. In some embodiments, a chiralally pure oligonucleotide composition comprises multiple oligonucleotides, where the oligonucleotides in this plurality are structurally identical and all have the same structure (same stereoisomer morphology; in the context of oligonucleotides, typically the same diastereomer morphology, as oligonucleotides typically have multiple chiral centers), and the chiralally pure oligonucleotide composition is not a diastereomer of any other stereoisomer (in the context of oligonucleotides, typically the same diastereomer, as oligonucleotides typically have multiple chiral centers; This does not include, for example, anything achievable by stereoselective preparation. As those skilled in the art will understand, sterically random (or "racemic," "uncontrolled chiral") oligonucleotide compositions have many stereoisomers (e.g., 2 n It is a random mixture of n diastereoisomers [wherein n is the number of chiral-binding phosphorus cells for oligonucleotides in which other chiral centers (e.g., carbon chiral centers of sugars) are chiral-controlled and exist independently in one configuration, and only the chiral-binding phosphorus center is not chiral-controlled].

[0287] Specific data demonstrating that chiral-controlled oligonucleotide compositions, such as chiral-controlled MAPT oligonucleotide compositions, are characterized and / or active in reducing the level, activity and / or expression of MAPT target genes or their gene products are shown, for example, in the Examples section of this document.

[0288] In some embodiments, the disclosure provides an oligonucleotide composition comprising an oligonucleotide containing at least one chiral-bound phosphorus. In some embodiments, the disclosure provides a MAPT oligonucleotide composition comprising a MAPT oligonucleotide containing at least one chiral-bound phosphorus. In some embodiments, the disclosure provides a MAPT oligonucleotide composition comprising a chiral-controlled phosphorothioate internucleotide bond, wherein the bound phosphorus has an Rp configuration. In some embodiments, the disclosure provides a MAPT oligonucleotide composition comprising a chiral-controlled phosphorothioate internucleotide bond, wherein the bound phosphorus has an Sp configuration.

[0289] In some embodiments, the provided chiral-controlled oligonucleotide composition (e.g., a chiral-controlled MAPT oligonucleotide composition) is surprisingly effective compared to a reference oligonucleotide composition. In some embodiments, the desired biological effect (e.g., when measured by the level of reduction of mRNA, protein, etc., which is the target of the reduction) can be enhanced by more than 5, 10, 15, 20, 25, 30, 40, 50, or 100 times (e.g., when measured by the level of residual mRNA, protein, etc.). In some embodiments, the change is measured by a decrease in undesirable mRNA levels compared to a reference condition. In some embodiments, the change is measured by an increase in desirable mRNA levels compared to a reference condition. In some embodiments, the change is measured by a decrease in undesirable mRNA levels compared to a reference condition. In some embodiments, the reference condition is, for example, the absence of treatment with the chiral-controlled oligonucleotide composition. In some embodiments, the reference condition is a corresponding sterically randomized composition of oligonucleotides having the same chemical composition.

[0290] In some embodiments, the disclosure provides chiral-controlled oligonucleotide compositions, for example, chiral-controlled MAPT oligonucleotide compositions, where at least one chiral-controlled internucleotide bond-binding phosphorus is Sp. In some embodiments, the disclosure provides chiral-controlled oligonucleotide compositions, for example, chiral-controlled MAPT oligonucleotide compositions, where the majority of chiral-controlled internucleotide bond-binding phosphorus is Sp. In some embodiments, Sp represents approximately 50%-100%, 55%-100%, 60%-100%, 65%-100%, 70%-100%, 75%-100%, 80%-100%, 85%-100%, 90%-100%, 55%-95%, 60%-95%, 65%-95%, or approximately 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or more of all chiral controlled phosphorothioate internucleotide bonds. In some embodiments, Sp represents approximately 50%-100%, 55%-100%, 60%-100%, 65%-100%, 70%-10% of all phosphorothioate internucleotide bonds. Chiral control is observed in 0%, 75%-100%, 80%-100%, 85%-100%, 90%-100%, 55%-95%, 60%-95%, 65%-95%, or approximately 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or above, and these are classified as Sp. In some embodiments, about 50% to 100%, 55% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 100%, 85% to 100%, 90% to 100%, 55% to 95%, 60% to 95%, 65% to 95%, or about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or more of all chiral-controlled internucleotide bonds (or all chiral internucleotide bonds, or all internucleotide bonds) are Sp. In some embodiments, the Disclosure provides chiral-controlled oligonucleotide compositions, for example, chiral-controlled MAPT oligonucleotide compositions, where the majority of chiral internucleotide bonds are chiral-controlled and their binding phosphorus is Sp. In some embodiments, the disclosure provides chiral-controlled oligonucleotide compositions, for example, chiral-controlled MAPT oligonucleotide compositions, where each chiral internucleotide bond is chiral-controlled and each chiral-linked phosphorus is Sp. In some embodiments, the disclosure provides chiral-controlled oligonucleotide compositions, for example, chiral-controlled MAPT oligonucleotide compositions, where at least one chiral-controlled internucleotide bond has Rp-linked phosphorus. In some embodiments, the disclosure provides chiral-controlled oligonucleotide compositions, for example, chiral-controlled MAPT oligonucleotide compositions, where at least one chiral-controlled internucleotide bond contains Rp-linked phosphorus and at least one chiral-controlled internucleotide bond contains Sp-linked phosphorus. In some embodiments, at least one phosphorothioate internucleotide bond is chiral-controlled and is Rp.In some embodiments, about 1 to 5, for example, about 1, 2, 3, 4, or 5 phosphorothioate internucleotide bonds are chiralized and are Rp. In some embodiments, about 50% to 100%, 55% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 100%, 85% to 100%, 90% to 100%, 55% to 95%, 60% to 95%, 65% to 95%, or about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or more of all chiralized non-negatively charged internucleotide bonds (e.g., n001) are Rp. In some embodiments, each chiralized n001 is Rp.

[0291] Stereochemistry and patterns of chiral centers in the skeleton In contrast to natural phosphate bonds, the phosphate groups in chiral-modified internucleotide bonds, such as phosphorothioate internucleotide bonds, are chiral. In particular, this disclosure provides techniques (e.g., oligonucleotides, compositions, methods, etc.) that involve controlling the stereochemistry of chiral phosphate groups in chiral internucleotide bonds. In some embodiments, as demonstrated herein, stereochemical control can result in improved properties and / or activity, including desired stability, reduced toxicity, and improved target nucleic acid reduction. In some embodiments, this disclosure provides a useful skeletal chiral center pattern for oligonucleotides and / or regions thereof, which is a combination of the stereochemistry (Rp or Sp) of each chiral phosphate group in the chiral phosphate group, an indication (Op, if present), etc., from 5' to 3'. In some embodiments, the skeletal chiral center pattern can control the cleavage pattern of a target nucleic acid when it comes into contact with the provided oligonucleotide or composition thereof in a cleavage system (e.g., in vitro assays, cells, tissues, organs, organisms, subjects, etc.). In some embodiments, the pattern of skeletal chiral centers improves the cleavage efficiency and / or selectivity of target nucleic acids when in contact with oligonucleotides or compositions thereof provided in the cleavage system.

[0292] In some embodiments, the chiral center pattern of the oligonucleotide or region thereof includes or is any (Np)n(Op)m (wherein Np is Rp or Sp, Op is achiral (as in the case of the bound phosphorus of a natural phosphate bond), and each of n and m is independently as defined and described herein). In some embodiments, the chiral center pattern of the oligonucleotide or region thereof includes or is (Sp)n(Op)m (wherein each variable is independently as defined and described herein). In some embodiments, the chiral center pattern of the oligonucleotide or region thereof includes or is (Rp)n(Op)m (wherein each variable is independently as defined and described herein). In some embodiments, n is 1. In some embodiments, the chiral center pattern of the oligonucleotide or region thereof comprises or includes (Sp)(Op)m (wherein m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, the chiral center pattern of the oligonucleotide or region thereof comprises or includes (Rp)(Op)m (wherein m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, the chiral center pattern of the 5'-wing skeleton is (Np)n(Op)m or includes it. In some embodiments, the chiral center pattern of the 5'-wing skeleton is (Sp)n(Op)m or includes it. In some embodiments, the chiral center pattern of the 5'-wing skeleton is (Rp)n(Op)m or includes it. In some embodiments, the chiral center pattern of the 5'-wing skeleton is (Sp)(Op)m or includes it. In some embodiments, the skeletal chiral center pattern of the 5'-wing is (Rp)(Op)m or includes it. In some embodiments, the skeletal chiral center pattern of the 5'-wing is (Sp)(Op)m. In some embodiments, the skeletal chiral center pattern of the 5'-wing is (Rp)(Op)m.In some embodiments, the 5'-wing skeletal chiral center pattern is (Sp)(Op)m, where Sp is the binding phosphorus configuration of the first internucleotide bond from the 5' end in the oligonucleotide. In some embodiments, the 5'-wing skeletal chiral center pattern is (Rp)(Op)m, where Rp is the binding phosphorus configuration of the first internucleotide bond from the 5' end in the oligonucleotide. In some embodiments, as described in this disclosure, m is 2; in some embodiments, m is 3; in some embodiments, m is 4; in some embodiments, m is 5; in some embodiments, m is 6.

[0293] In some embodiments, the chiral center pattern of the oligonucleotide or region thereof comprises or is (Op)m(Np)n (wherein Np is Rp or Sp, Op indicates that the bound phosphorus is achiral (for example, as in the case of bound phosphorus of a natural phosphate bond), and each of n and m is independently as defined and described herein). In some embodiments, the chiral center pattern of the oligonucleotide or region thereof comprises or is (Op)m(Sp)n (wherein each variable is independently as defined and described herein). In some embodiments, the chiral center pattern of the oligonucleotide or region thereof comprises or is (Op)m(Rp)n (wherein each variable is independently as defined and described herein). In some embodiments, n is 1. In some embodiments, the chiral center pattern of the oligonucleotide or region thereof comprises or is (Op)m(Sp) (wherein m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, the chiral center pattern of the oligonucleotide or region thereof comprises or is (Op)m(Rp) (wherein m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, the chiral center pattern of the 3'-wing skeleton is (Op)m(Np)n or It includes it. In some embodiments, the skeletal chiral center pattern of the 3'-wing is (Op)m(Rp)n or includes it. In some embodiments, the skeletal chiral center pattern of the 3'-wing is (Op)m(Sp) or includes it. In some embodiments, the skeletal chiral center pattern of the 3'-wing is (Op)m(Rp) or includes it. In some embodiments, the skeletal chiral center pattern of the 3'-wing is (Op)m(Sp). In some embodiments, the skeletal chiral center pattern of the 3'-wing is (Op)m(Rp). In some embodiments, the skeletal chiral center pattern of the 3'-wing is (Op)m(Sp), where Sp is the binding phosphorus configuration of the 5' end to the last internucleotide bond in the oligonucleotide. In some embodiments, the skeletal chiral center pattern of the 3'-wing is (Op)m(Rp), where Rp is the binding phosphorus configuration of the 5' end to the last internucleotide bond in the oligonucleotide. In some embodiments, m is 2 as described in this disclosure; in some embodiments, m is 3; in some embodiments, m is 4; in some embodiments, m is 5; in some embodiments, m is 6.

[0294] In some embodiments, the skeletal chiral center pattern of an oligonucleotide or its region (e.g., the core) includes or is (Sp)m(Rp / Op)n or (Rp / Op)n(Sp)m (wherein each variable is independently as described in this disclosure). In some embodiments, the skeletal chiral center pattern of an oligonucleotide or its region (e.g., the core) includes or is (Sp)m(Rp)n or (Rp)n(Sp)m (wherein each variable is independently as described in this disclosure). In some embodiments, the skeletal chiral center pattern of an oligonucleotide or its region (e.g., the core) includes or is (Sp)m(Op)n or (Op)n(Sp)m (wherein each variable is independently as described in this disclosure). In some embodiments, the skeletal chiral center pattern of an oligonucleotide or region thereof (e.g., the core) includes or is (Np)t[(Rp / Op)n(Sp)m]y or [(Rp / Op)n(Sp)m]y(Np)t (wherein y is 1 to 50, and each of the other variables is independently as described in this disclosure). In some embodiments, the chiral center pattern of the oligonucleotide or its region (e.g., core) includes or is [(Rp / Op)n(Sp)m]y(Rp)k, [(Rp / Op)n(Sp)m]y, (Sp)t[(Rp / Op)n(Sp)m]y, (Sp)t[(Rp / Op)n(Sp)m]y(Rp)k (wherein k is 1 to 50, and each of the other variables is independently as described herein). In some embodiments, the chiral center pattern of the oligonucleotide or its region (e.g., core) comprises or is [(Op)n(Sp)m]y(Rp)k, [(Op)n(Sp)m]y, (Sp)t[(Op)n(Sp)m]y, (Sp)t[(Op)n(Sp)m]y(Rp)k (wherein each variable is independently as described herein).In some embodiments, the chiral center pattern of the oligonucleotide or its region (e.g., core) includes or is [(Rp)n(Sp)m]y(Rp)k, [(Rp)n(Sp)m]y, (Sp)t[(Rp)n(Sp)m]y, (Sp)t[(Rp)n(Sp)m]y(Rp)k (wherein each variable is independently as described herein). In some embodiments, the oligonucleotide includes a core region. In some embodiments, the oligonucleotide includes a core region where each sugar in the core region is 2'-OR. 1 (In the formula, R 1 (as described in this disclosure) is not included. In some embodiments, the oligonucleotide comprises a core region, where each sugar in the core region is independently a native DNA sugar. In some embodiments, the core's skeletal chiral center pattern comprises or is (Rp)(Sp)m. In some embodiments, the core's skeletal chiral center pattern comprises (Op)(Sp)m. In some embodiments, the core's skeletal chiral center pattern includes or is (Np)t[(Rp / Op)n(Sp)m]y or [(Rp / Op)n(Sp)m]y(Np)t. In some embodiments, the core's skeletal chiral center pattern includes or is (Np)t[(Rp / Op)n(Sp)m]y or [(Rp / Op)n(Sp)m]y(Np)t. In some embodiments, the core's skeletal chiral center pattern includes or is (Np)t[(Rp)n(Sp)m]y or [(Rp)n(Sp)m]y(Np)t. In some embodiments, the core's skeletal chiral center pattern includes or is [(Rp / Op)n(Sp)m]y(Rp)k, [(Rp / Op)n(Sp)m]y, (Sp)t[(Rp / Op)n(Sp)m]y, (Sp)t[(Rp / Op)n(Sp)m]y(Rp)k. In some embodiments, the core's skeletal chiral center pattern includes or is [(Op)n(Sp)m]y(Rp)k, [(Op)n(Sp)m]y, (Sp)t[(Op)n(Sp)m]y, (Sp)t[(Op)n(Sp)m]y(Rp)k. In some embodiments, the core's skeletal chiral center pattern includes or is [(Rp)n(Sp)m]y(Rp)k, [(Rp)n(Sp)m]y, (Sp)t[(Rp)n(Sp)m]y, or (Sp)t[(Rp)n(Sp)m]y(Rp)k. In some embodiments, the core's skeletal chiral center pattern includes [(Rp)n(Sp)m]y(Rp)k. In some embodiments, the core's skeletal chiral center pattern includes [(Rp)n(Sp)m]y(Rp). In some embodiments, the core's skeletal chiral center pattern includes [(Rp)n(Sp)m]y. In some embodiments, the core's skeletal chiral center pattern includes (Sp)t[(Rp)n(Sp)m]y. In some embodiments, the core's skeletal chiral center pattern includes (Sp)t[(Rp)n(Sp)m]y(Rp)k. In some embodiments, the core's skeletal chiral center pattern includes (Sp)t[(Rp)n(Sp)m]y(Rp). In some embodiments, the core's skeletal chiral center pattern is [(Rp)n(Sp)m]y(Rp)k.In some embodiments, the core's skeletal chiral center pattern is [(Rp)n(Sp)m]y(Rp). In some embodiments, the core's skeletal chiral center pattern is [(Rp)n(Sp)m]y. In some embodiments, the core's skeletal chiral center pattern is (Sp)t[(Rp)n(Sp)m]y. In some embodiments, the core's skeletal chiral center pattern is (Sp)t[(Rp)n(Sp)m]y(Rp)k. In some embodiments, the core's skeletal chiral center pattern is (Sp)t[(Rp)n(Sp)m]y(Rp). In some embodiments, each n is 1. In some embodiments, each t is 1. In some embodiments, t is 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, each of t and n is 1. In some embodiments, each m is 2 or greater. In some embodiments, k is 1. In some embodiments, k is 2 to 10.

[0295] In some embodiments, the skeletal chiral center pattern includes or is (Sp)m(Rp)n, (Rp)n(Sp)m, (Np)t(Rp)n(Sp)m, (Sp)t(Rp)n(Sp)m, (Np)t[(Rp)n(Sp)m]2, (Sp)t[(Rp)n(Sp)m]2, (Np)t(Op)n(Sp)m, (Sp)t(Op)n(Sp)m, (Np)t[(Op)n(Sp)m]2, or (Sp)t[(Op)n(Sp)m]2. In some embodiments, the pattern is (Np)t(Op / Rp)n(Sp)m(Op / Rp)n(Sp)m. In some embodiments, the pattern is (Np)t(Op / Rp)n(Sp)1~5(Op / Rp)n(Sp)m. In some embodiments, the pattern is (Np)t(Op / Rp)n(Sp)2~5(Op / Rp)n(Sp)m. In some embodiments, the pattern is (Np)t(Op / Rp)n(Sp)2(Op / Rp)n(Sp)m. In some embodiments, the pattern is (Np)t(Op / Rp)n(Sp)3(Op / Rp)n(Sp)m. In some embodiments, the pattern is (Np)t(Op / Rp)n(Sp)4(Op / Rp)n(Sp)m. In some embodiments, the pattern is (Np)t(Op / Rp)n(Sp)5 (Op / Rp)n(Sp)m

[0296] In some embodiments, Np is Sp. In some embodiments, (Op / Rp) is Op. In some embodiments, (Op / Rp) is Rp. In some embodiments, Np is Sp and (Op / Rp) is Rp. In some embodiments, Np is Sp and (Op / Rp) is Op. In some embodiments, Np is Sp and at least one (Op / Rp) is Rp and at least one (Op / Rp) is Op. In some embodiments, the skeletal chiral center pattern includes or is (Rp)n(Sp)m, (Np)t(Rp)n(Sp)m, or (Sp)t(Rp)n(Sp)m (wherein m>2). In some embodiments, the skeletal chiral center pattern includes or is (Rp)n(Sp)m, (Np)t(Rp)n(Sp)m, or (Sp)t(Rp)n(Sp)m (wherein n is 1, at least one t>1, and at least one m>2).

[0297] In some embodiments, oligonucleotides containing a core region whose skeletal chiral center pattern begins with Rp can provide high activity and / or improved properties. In some embodiments, oligonucleotides containing a core region whose skeletal chiral center pattern ends with Rp can provide high activity and / or improved properties. In some embodiments, oligonucleotides containing a core region whose skeletal chiral center pattern begins with Rp can provide high activity (e.g., targeted cleavage) without significantly affecting their properties, such as stability. In some embodiments, oligonucleotides containing a core region whose skeletal chiral center pattern ends with Rp can provide high activity (e.g., targeted cleavage) without significantly affecting their properties, such as stability. In some embodiments, the skeletal chiral center pattern begins with Rp and ends with Sp. In some embodiments, the skeletal chiral center pattern begins with Rp and ends with Rp. In some embodiments, the skeletal chiral center pattern begins with Sp and ends with Rp. Typically, with respect to the skeletal chiral center pattern, internucleotide bonds linking core nucleosides and wing nucleosides are included in the core region pattern. In many embodiments as described in this disclosure (e.g., the various oligonucleotides in Table 1), the wing sugar linked by such internucleotide linkages has a different structure from the core sugar linked by the same internucleotide linkage (e.g., in some embodiments, the wing sugar includes a 2'-modification, while the core sugar does not include the same 2'-modification or has two -H at the 2' position). In some embodiments, the wing sugar includes a sugar modification that the core sugar does not. In some embodiments, the wing sugar is a modified sugar, while the core sugar is a natural DNA sugar. In some embodiments, the wing sugar includes a sugar modification at the 2' position (less than two -H at the 2' position), while the core sugar does not have a modification at the 2' position (two -H at the 2' position).

[0298] In some embodiments, as demonstrated herein, an additional Rp internucleotide bond connects a sugar without a 2'-substituent (e.g., core sugar) to a sugar with a 2'-modification (e.g., 2'-OR', where R' is optionally substituted C). 1~6 It is an aliphatic sugar (e.g., 2'-OMe, 2'-MOE, etc.), which may be a wing sugar) that is linked to the other sugar. In some embodiments, an internucleotide linkage in which a sugar without a 2'-substituent is linked to the 5' end (e.g., to the 3'-carbon of the sugar) and a sugar containing a 2'-modification is linked to the 3' end (e.g., to the 5'-carbon of the sugar) is an Rp internucleotide linkage. In some embodiments, an internucleotide linkage in which a sugar without a 2'-substituent is linked to the 3' end (e.g., to the 5'-carbon of the sugar) and a sugar containing a 2'-modification is linked to the 5' end (e.g., to the 3'-carbon of the sugar) is an Rp internucleotide linkage. In some embodiments, each internucleotide linkage linking a sugar without a 2'-substituent and a sugar containing a 2'-modification is independently an Rp internucleotide linkage. In some embodiments, the Rp internucleotide linkage is an Rp phosphorothioate internucleotide linkage.

[0299] In some embodiments, the skeletal chiral center pattern of a MAPT oligonucleotide or its region (e.g., core) includes or is (Op)[(Rp / Op)n(Sp)m]y(Rp)k(Op), (Op)[(Rp / Op)n(Sp)m]y(Op), (Op)(Sp)t[(Rp / Op)n(Sp)m]y(Op), or (Op)(Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op) (wherein k is 1 to 50, and each of the other variables is independently as described herein). In some embodiments, the chiral center pattern of the MAPT oligonucleotide skeleton is (Op)[(Rp / Op)n(Sp)m]y(Rp)k(Op), (Op)[(Rp / Op)n(Sp)m]y(Op), (Op)(Sp)t[(Rp / Op)n(Sp)m]y(Op), or (Op)(Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op) (wherein f, g, h and j are independently 1 to 50, and Each of the other variables is independently as described in this disclosure, and the oligonucleotide comprises a core region whose skeletal chiral center pattern comprises or includes [(Rp / Op)n(Sp)m]y(Rp)k, [(Rp / Op)n(Sp)m]y, (Sp)t[(Rp / Op)n(Sp)m]y, or (Sp)t[(Rp / Op)n(Sp)m]y(Rp)k as described in this disclosure. In some embodiments, the skeletal chiral center pattern is or includes (Op)[(Rp / Op)n(Sp)m]y(Rp)k(Op). In some embodiments, the skeletal chiral center pattern is or includes (Op)[(Rp / Op)n(Sp)m]y(Rp)(Op). In some embodiments, the skeletal chiral center pattern is (Op)[(Rp / Op)n(Sp)m]y(Op) or includes the same. In some embodiments, the skeletal chiral center pattern is (Op)(Sp)t[(Rp / Op)n(Sp)m]y(Op) or includes the same. In some embodiments, the skeletal chiral center pattern is (Op)(Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op) or includes the same.In some embodiments, the skeletal chiral center pattern is (Op)(Sp)t[(Rp / Op)n(Sp)m]y(Rp)(Op) or includes the same. In some embodiments, the skeletal chiral center pattern is (Op)[(Rp)n(Sp)m]y(Rp)k(Op) or includes the same. In some embodiments, the skeletal chiral center pattern is (Op)[(Rp)n(Sp)m]y(Rp)(Op) or includes the same. In some embodiments, the skeletal chiral center pattern is (Op)[(Rp)n(Sp)m]y(Op) or includes the same. In some embodiments, the skeletal chiral center pattern is (Op)(Sp)t[(Rp)n(Sp)m]y(Op) or includes the same. In some embodiments, the skeletal chiral center pattern is (Op)(Sp)t[(Rp)n(Sp)m]y(Rp)k(Op) or includes the same. In some embodiments, the skeletal chiral center pattern is (Op)(Sp)t[(Rp)n(Sp)m]y(Rp)(Op) or includes the same. In some embodiments, each n is 1. In some embodiments, k is 1. In some embodiments, k is 2 to 10.

[0300] In some embodiments, the skeletal chiral center pattern of a MAPT oligonucleotide or its region (e.g., core) includes or is (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j, (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Op)h(Np)j, (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Op)h(Np)j, or (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j (wherein each of f, g, h, and j is independently 1 to 50, and each of the other variables is independently as described herein). In some embodiments, the chiral center pattern of the MAPT oligonucleotide skeleton is (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j, (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Op)h(Np)j, (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp) The oligonucleotide comprises or includes m]y(Op)h(Np)j, or (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j, and the oligonucleotide comprises a core region having a skeletal chiral center pattern as described herein, comprising or including [(Rp / Op)n(Sp)m]y(Rp)k, [(Rp / Op)n(Sp)m]y, (Sp)t[(Rp / Op)n(Sp)m]y, or (Sp)t[(Rp / Op)n(Sp)m]y(Rp)k. In some embodiments, the chiral center pattern of the MAPT oligonucleotide skeleton is (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j, (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Op)h(Np)j, (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Op)h(Np)j, or (Np)f(Op)g(Sp)t[(R The oligonucleotide is p / Op)n(Sp)m]y(Rp)k(Op)h(Np)j, and this oligonucleotide includes a core region having a skeletal chiral center pattern of [(Rp / Op)n(Sp)m]y(Rp)k, [(Rp / Op)n(Sp)m]y, (Sp)t[(Rp / Op)n(Sp)m]y, or (Sp)t[(Rp / Op)n(Sp)m]y(Rp)k as described herein. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Rp)(Op)h(Np)j or includes it. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Op)h(Np)j or includes it. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Op)h(Np)j or includes it. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j or includes it.In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Rp)(Op)h(Np)j or includes the same. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g[(Rp)n(Sp)m]y(Rp)k(Op)h(Np)j or includes the same. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g[(Rp)n(Sp)m]y(Rp)(Op)h(Np)j or includes the same. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g[(Rp)n(Sp)m]y(Op)h(Np)j or includes the same. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g(Sp)t[(Rp)n(Sp)m]y(Op)h(Np)j or includes the same. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g(Sp)t[(Rp)n(Sp)m]y(Rp)k(Op)h(Np)j or includes the same. In some embodiments, the skeletal chiral center pattern is (Np)f(Op)g(Sp)t[(Rp)n(Sp)m]y(Rp)(Op)h(Np)j or includes the same. In some embodiments, at least one Np is Sp. In some embodiments, at least one Np is Rp. In some embodiments, the Np furthest 5' is Sp. In some embodiments, the Np furthest 3' is Sp. In some embodiments, each Np is Sp. In some embodiments, (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j is (Sp)(Op)g[(Rp)n(Sp)m]y(Rp)k(Op)h(Sp). In some embodiments, (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j is (Sp)(Op)g[(Rp)n(Sp)m]y(Rp)(Op)h(Sp). In some embodiments, the chiral center pattern of the oligonucleotide skeleton is (Sp)(Op)g[(Rp)n(Sp)m]y(Rp)(Op)h(Sp) or includes it.In some embodiments, the chiral center pattern of the oligonucleotide skeleton is (Sp)(Op)g[(Rp)n(Sp)m]y(Rp)(Op)h(Sp). In some embodiments, (Np)f(Op)g[(Rp / Op)n(Sp)m]y(Op)h(Np)j is (Sp)(Op)g[(Rp)n(Sp)m]y(Op)h(Sp). In some embodiments, the chiral center pattern of the oligonucleotide skeleton is (Sp)(Op)g[(Rp)n(Sp)m]y(Op)h(Sp) or includes it. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is (Sp)(Op)g[(Rp)n(Sp)m]y(Op)h(Sp). In some embodiments, (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Op)h(Np)j is (Sp)(Op)g(Sp)t[(Rp)n(Sp)m]y(Op)h(Sp). In some embodiments, the chiral center pattern of the oligonucleotide skeleton is (Sp)(Op)g(Sp)t[(Rp)n(Sp)m]y(Op)h(Sp) or includes it. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is (Sp)(Op)g(Sp)t[(Rp)n(Sp)m]y(Op)h(Sp). In some embodiments, (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j is (Sp)(Op)g(Sp)t[(Rp)n(Sp)m]y(Rp)k(Op)h(Sp). In some embodiments, (Np)f(Op)g(Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j is (Sp)(Op)g(Sp)t[(Rp)n(Sp)m]y(Rp)(Op)h(Sp). In some embodiments, the chiral center pattern of the oligonucleotide skeleton is (Sp)(Op)g(Sp)t[(Rp)n(Sp)m]y(Rp)(Op)h(Sp). In some embodiments, the chiral center pattern of the oligonucleotide skeleton is (Sp)(Op)g(Sp)t[(Rp)n(Sp)m]y(Rp)(Op)h(Sp). In some embodiments, each n is 1. In some embodiments, f is 1. In some embodiments, g is 1. In some embodiments, g is greater than 1. In some embodiments, g is 2. In some embodiments, g is 3. In some embodiments, g is 4.In some embodiments, g is 5. In some embodiments, g is 6. In some embodiments, g is 7. In some embodiments, g is 8. In some embodiments, g is 9. In some embodiments, g is 10. In some embodiments, h is 1. In some embodiments, h is greater than 1. In some embodiments, h is 2. In some embodiments, h is 3. In some embodiments, h is 4. In some embodiments, h is 5. In some embodiments, h is 6. In some embodiments, h is 7. In some embodiments, h is 8. In some embodiments, h is 9. In some embodiments, h is 10. In some embodiments, j is 1. In some embodiments, k is 1. In some embodiments, k is between 2 and 10.

[0301] In some embodiments, the chiral center pattern of the MAPT oligonucleotide or its region (e.g., core) includes or is comprised of [(Rp / Op)n(Sp)m]y, (Sp)t[(Rp / Op)n(Sp)m]y, (Sp)t[(Rp / Op)n(Sp)m]yRp, [(Rp / Op)n(Sp)m]y(Rp)k, (Sp)t[(Rp / Op)n(Sp)m]y(Rp)k, (Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op)h, and (Sp)t[(Rp / Op)n(Sp)m]y(Rp)k(Op)h(Np)j (wherein each variable is independently as described herein).

[0302] In some embodiments, in the provided skeletal chiral center pattern, at least one (Rp / Op) is Rp. In some embodiments, at least one (Rp / Op) is Op. In some embodiments, each (Rp / Op) is Rp. In some embodiments, each (Rp / Op) is Op. In some embodiments, at least one of the [(Rp)n(Sp)m]y or [(Rp / Op)n(Sp)m]y of the pattern is RpSp. In some embodiments, the [(Rp)n(Sp)m]y or [ At least one of (Rp / Op)n(Sp)m]y is RpSpSp or includes it. In some embodiments, at least one of [(Rp)n(Sp)m]y or [(Rp / Op)n(Sp)m]y in the pattern is RpSp, and at least one of [(Rp)n(Sp)m]y or [(Rp / Op)n(Sp)m]y in the pattern is RpSpSp or includes it. For example, in some embodiments, [(Rp)n(Sp)m]y in the pattern is (RpSp)[(Rp)n(Sp)m] (y-1) And; in some embodiments, [(Rp)n(Sp)m]y in the pattern is (RpSp)[RpSpSp(Sp) (m-2) ][(Rp)n(Sp)m] (y-2) In some embodiments, (Sp)t[(Rp)n(Sp)m]y(Rp) is (Sp)t(RpSp)[(Rp)n(Sp)m] (y-1) (Rp) is. In some embodiments, (Sp)t[(Rp)n(Sp)m]y(Rp) is (Sp)t(RpSp)[RpSpSp(Sp) (m-2) ][(Rp)n(Sp)m] (y-2) (Rp) is the case. In some embodiments, each [(Rp / Op)n(Sp)m] is independently [Rp(Sp)m]. In some embodiments, the first Sp of (Sp)t represents the binding phosphorus stereochemistry of the first internucleotide bond of the oligonucleotide from 5' to 3'. In some embodiments, the first Sp of (Sp)t represents the binding phosphorus stereochemistry of the first internucleotide bond of a region from 5' to 3', e.g., the core region. In some embodiments, the last Np of (Np)j represents the binding phosphorus stereochemistry of the last internucleotide bond of the oligonucleotide from 5' to 3'. In some embodiments, the last Np is Sp.

[0303] In some embodiments, the skeletal chiral center pattern of the oligonucleotide or region (e.g., the 5'-wing region) is or includes Sp(Op)3. In some embodiments, the skeletal chiral center pattern of the oligonucleotide or region (e.g., the 5'-wing region) is or includes Rp(Op)3. In some embodiments, the skeletal chiral center pattern of the oligonucleotide or region (e.g., the 3'-wing region) is (Op)3Sp. In some embodiments, the skeletal chiral center pattern of the oligonucleotide or region (e.g., the 3'-wing region) is (Op)3Rp. In some embodiments, the skeletal chiral center pattern of the oligonucleotide or region (e.g., the 3'-wing region) is (Op)3Rp. In some embodiments, the skeletal chiral center pattern of the oligonucleotide or region (e.g., the core region) is Rp(Sp)4Rp(Sp)4Rp. In some embodiments, the skeletal chiral center pattern of the oligonucleotide or region (e.g., the core region) is (Sp)5Rp(Sp)4Rp. In some embodiments, the skeletal chiral center pattern of the oligonucleotide or region (e.g., the core region) is (Sp)5Rp(Sp)5 or includes it. In some embodiments, the skeletal chiral center pattern of the oligonucleotide or region (e.g., the core region) is Rp(Sp)4Rp(Sp)5 or includes it. In some embodiments, the skeletal chiral center pattern of the oligonucleotide is Np(Op)3Rp(Sp)4Rp(Sp)4Rp(Op)3Np or includes it. In some embodiments, the skeletal chiral center pattern of the oligonucleotide is Np(Op)3(Sp)5Rp(Sp)4Rp(Op)3Np or includes it. In some embodiments, the skeletal chiral center pattern of the oligonucleotide is Np(Op)3(Sp)5Rp(Sp)5(Op)3Np or includes it. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is or includes Np(Op)3Rp(Sp)4Rp(Sp)5(Op)3Np. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is or includes Sp(Op)3Rp(Sp)4Rp(Sp)4Rp(Op)3Sp.In some embodiments, the chiral center pattern of the oligonucleotide skeleton is or includes Sp(Op)3(Sp)5Rp(Sp)4Rp(Op)3Sp. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is or includes Sp(Op)3(Sp)5Rp(Sp)5(Op)3Sp. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is. , Sp(Op)3Rp(Sp)4Rp(Sp)5(Op)3Sp or includes the same. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is Rp(Op)3Rp(Sp)4Rp(Sp)4Rp(Op)3Rp or includes the same. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is Rp(Op)3(Sp)5Rp(Sp)4Rp(Op)3Rp or includes the same. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is Rp(Op)3(Sp)5Rp(Sp)5(Op)3Rp or includes the same. In some embodiments, the chiral center pattern of the oligonucleotide skeleton is Rp(Op)3Rp(Sp)4Rp(Sp)5(Op)3Rp or includes the same.

[0304] In some embodiments, each of m, y, t, n, k, f, g, h, and j is independently 1 to 25.

[0305] In some embodiments, m is 1 to 25. In some embodiments, m is 1 to 20. In some embodiments, m is 1 to 15. In some embodiments, m is 1 to 10. In some embodiments, m is 1 to 5. In some embodiments, m is 2 to 20. In some embodiments, m is 2 to 15. In some embodiments, m is 2 to 10. In some embodiments, m is 2 to 5. In some embodiments, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, in the skeletal chiral center pattern, each m is independently 2 or more. In some embodiments, each m is independently 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, each m is independently 2-3, 2-5, 2-6, or 2-10. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments, m is 5. In some embodiments, m is 6. In some embodiments, m is 7. In some embodiments, m is 8. In some embodiments, m is 9. In some embodiments, m is 10. In some embodiments, if m appears more than once, they may be the same or different, and each of them independently is as described in this disclosure.

[0306] In some embodiments, y is 1 to 25. In some embodiments, y is 1 to 20. In some embodiments, y is 1 to 15. In some embodiments, y is 1 to 10. In some embodiments, y is 1 to 5. In some embodiments, y is 2 to 20. In some embodiments, y is 2 to 15. In some embodiments, y is 2 to 10. In some embodiments, y is 2 to 5. In some embodiments, y is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, y is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, y is 1. In some embodiments, y is 2. In some embodiments, y is 3. In some embodiments, y is 4. In some embodiments, y is 5. In some embodiments, y is 6. In some embodiments, y is 7. In some embodiments, y is 8. In some embodiments, y is 9. In some embodiments, y is 10.

[0307] In some embodiments, t is 1 to 25. In some embodiments, t is 1 to 20. In some embodiments, t is 1 to 15. In some embodiments, t is 1 to 10. In some embodiments, t is 1 to 5. In some embodiments, t is 2 to 20. In some embodiments, t is 2 to 15. In some embodiments, t is 2 to 10. In some embodiments, t is 2 to 5. In some embodiments, t is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, each t is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, t is 2 or more. In some embodiments, t is 1. In some embodiments, t is 2. In some embodiments, t is 3. In some embodiments, t is 4. In some embodiments, t is 5. In some embodiments, t is 6. In some embodiments, t is 7. In some embodiments, t is 8. In some embodiments, t is 9. In some embodiments, t is 10. In some embodiments, if t appears more than once, they may be the same or different, and each of them independently is as described in this disclosure.

[0308] In some embodiments, n is 1 to 25. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. In some embodiments, if n appears more than once, they may be the same or different, and each of them may be independently as described in this disclosure. In many embodiments, in the skeletal chiral center pattern, at least one appearance of n is 1; in some cases, each n is 1.

[0309] In some embodiments, k is 1 to 25. In some embodiments, k is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, k is 1. In some embodiments, k is 2. In some embodiments, k is 3. In some embodiments, k is 4. In some embodiments, k is 5. In some embodiments, k is 6. In some embodiments, k is 7. In some embodiments, k is 8. In some embodiments, k is 9. In some embodiments, k is 10.

[0310] In some embodiments, f is 1 to 25. In some embodiments, f is 1 to 20. In some embodiments, f is 1 to 10. In some embodiments, f is 1 to 5. In some embodiments, f is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, f is 1. In some embodiments, f is 2. In some embodiments, f is 3. In some embodiments, f is 4. In some embodiments, f is 5. In some embodiments, f is 6. In some embodiments, f is 7. In some embodiments, f is 8. In some embodiments, f is 9. In some embodiments, f is 10.

[0311] In some embodiments, g is 1 to 25. In some embodiments, g is 1 to 20. In some embodiments, g is 1 to 10. In some embodiments, g is 1 to 5. In some embodiments, g is 2 to 10. In some embodiments, g is 2 to 5. In some embodiments, g is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, g is 1. In some embodiments In some embodiments, g is 2. In some embodiments, g is 3. In some embodiments, g is 4. In some embodiments, g is 5. In some embodiments, g is 6. In some embodiments, g is 7. In some embodiments, g is 8. In some embodiments, g is 9. In some embodiments, g is 10.

[0312] In some embodiments, h is 1 to 25. In some embodiments, h is 1 to 10. In some embodiments, h is 1 to 5. In some embodiments, h is 2 to 10. In some embodiments, h is 2 to 5. In some embodiments, h is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, h is 1. In some embodiments, h is 2. In some embodiments, h is 3. In some embodiments, h is 4. In some embodiments, h is 5. In some embodiments, h is 6. In some embodiments, h is 7. In some embodiments, h is 8. In some embodiments, h is 9. In some embodiments, h is 10.

[0313] In some embodiments, j is 1 to 25. In some embodiments, j is 1 to 10. In some embodiments, j is 1 to 5. In some embodiments, j is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25. In some embodiments, j is 1. In some embodiments, j is 2. In some embodiments, j is 3. In some embodiments, j is 4. In some embodiments, j is 5. In some embodiments, j is 6. In some embodiments, j is 7. In some embodiments, j is 8. In some embodiments, j is 9. In some embodiments, j is 10.

[0314] In some embodiments, at least one n is 1 and at least one m is 2 or greater. In some embodiments, at least one n is 1, at least one t is 2 or greater, and at least one m is 3 or greater. In some embodiments, each n is 1. In some embodiments, t is 1. In some embodiments, at least one t>1. In some embodiments, at least one t>2. In some embodiments, at least one t>3. In some embodiments, at least one t>4. In some embodiments, at least one m>1. In some embodiments, at least one m>2. In some embodiments, at least one m>3. In some embodiments, at least one m>4. In some embodiments, the skeletal chiral center pattern contains one or more achiral native phosphate bonds. In some embodiments, the sum of m, t, and n (or m and n if t is not present in the pattern) is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or greater. In some embodiments, the sum is 5. In some embodiments, the sum is 6. In some embodiments, the sum is 7. In some embodiments, the sum is 8. In some embodiments, the sum is 9. In some embodiments, the sum is 10. In some embodiments, the sum is 11. In some embodiments, the sum is 12. In some embodiments, the sum is 13. In some embodiments, the sum is 14. In some embodiments, the sum is 15.

[0315] In some embodiments, several bound phosphate groups in the chiral-controlled internucleotide bond are Sp. In some embodiments, at least 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the chiral-controlled internucleotide bond are Sp. It contains phosphorus. In some embodiments, at least 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of all chiral internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of all internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, the percentage is at least 20%. In some embodiments, the percentage is at least 30%. In some embodiments, the percentage is at least 40%. In some embodiments, the percentage is at least 50%. In some embodiments, the percentage is at least 60%. In some embodiments, the percentage is at least 65%. In some embodiments, the percentage is at least 70%. In some embodiments, the percentage is at least 75%. In some embodiments, the percentage is at least 80%. In some embodiments, the percentage is at least 90%. In some embodiments, the percentage is at least 95%. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least 5 internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least six internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least seven internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus.In some embodiments, at least eight internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least nine internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least ten internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least eleven internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least twelve internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least thirteen internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least fourteen internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least fifteen internucleotide bonds are chiral-controlled internucleotide bonds having Sp-linked phosphorus. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 internucleotide bonds are chiral-controlled internucleotide bonds having Rp-binding phosphorus. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 or fewer internucleotide bonds are chiral-controlled internucleotide bonds having Rp-binding phosphorus. In some embodiments, one and not more than one internucleotide bonds in an oligonucleotide are chiral-controlled internucleotide bonds having Rp-binding phosphorus. In some embodiments, two and not more than two internucleotide bonds in an oligonucleotide are chiral-controlled internucleotide bonds having Rp-binding phosphorus.In some embodiments, internucleotide bonds of three and not more than three in an oligonucleotide. However, these are chiral-controlled internucleotide bonds having Rp-binding phosphorus. In some embodiments, four and not more than four internucleotide bonds in an oligonucleotide are chiral-controlled internucleotide bonds having Rp-binding phosphorus. In some embodiments, five and not more than five internucleotide bonds in an oligonucleotide are chiral-controlled internucleotide bonds having Rp-binding phosphorus.

[0316] In some embodiments, all, essentially all, or most of the internucleotide bonds in the oligonucleotide are in the Sp configuration, except that one or a few internucleotide bonds (e.g., 1, 2, 3, 4, or 5 of all chiral-controlled internucleotide bonds, or all chiral internucleotide bonds, or all internucleotide bonds less than 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5%) are in the Rp configuration. For example, all chiral-controlled internucleotide bonds in an oligonucleotide, or all chiral internucleotide bonds, or about 50% to 100%, 55% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 100%, 85% to 100%, 90% to 100%, 55% to 95%, 60% to 95%, 65% to 95%, or about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or more. In some embodiments, all, essentially all, or most of the internucleotide bonds in the core are in the Sp configuration, except for one or a few internucleotide bonds (e.g., 1, 2, 3, 4, or 5 of all chiral-controlled internucleotide bonds in the core, or all chiral internucleotide bonds, or all internucleotide bonds less than 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5%) which are in the Rp configuration (e.g., (For example, all chiral-controlled internucleotide bonds in the core, or all chiral internucleotide bonds, or about 50% to 100%, 55% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 100%, 85% to 100%, 90% to 100%, 55% to 95%, 60% to 95%, 65% to 95%, or about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or more).In some embodiments, all, essentially all, or most of the internucleotide bonds in the core are Sp-configured phosphorothioates, except that one or a few internucleotide bonds (e.g., 1, 2, 3, 4, or 5 of all chiral-controlled internucleotide bonds in the core, or all chiral internucleotide bonds, or all internucleotide bonds less than 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, or 5%) are Rp-configured phosphorothioates. (For example, all chiral controlled internucleotide bonds in the core, or all chiral internucleotide bonds, or about 50% to 100%, 55% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 100%, 85% to 100%, 90% to 100%, 55% to 95%, 60% to 95%, 65% to 95%, or about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, or more). In some embodiments, each internucleotide bond in the core is a phosphorothioate in the Sp configuration, with the exception of one phosphorothioate in the Rp configuration. In some embodiments, each internucleotide bond in the core is a phosphorothioate in the Sp configuration, with the exception of one phosphorothioate in the Rp configuration.

[0317] In some embodiments, the oligonucleotide contains one or more Rp internucleotide bonds. In some embodiments, the oligonucleotide contains one and more than one. Contains no Rp internucleotide bonds. In some embodiments, the oligonucleotide contains two or more Rp internucleotide bonds. In some embodiments, the oligonucleotide contains three or more Rp internucleotide bonds. In some embodiments, the oligonucleotide contains four or more Rp internucleotide bonds. In some embodiments, the oligonucleotide contains five or more Rp internucleotide bonds. In some embodiments, about 5% to 50% of all chiral-controlled internucleotide bonds in the oligonucleotide are Rp. In some embodiments, about 5% to 40% of all chiral-controlled internucleotide bonds in the oligonucleotide are Rp. In some embodiments, about 10% to 40% of all chiral-controlled internucleotide bonds in the oligonucleotide are Rp. In some embodiments, about 15% to 40% of all chiral-controlled internucleotide bonds in the oligonucleotide are Rp. In some embodiments, about 20% to 40% of all chiral-controlled internucleotide bonds in the oligonucleotide are Rp. In some embodiments, about 25% to 40% of all chiral-controlled internucleotide bonds in the oligonucleotide are Rp. In some embodiments, approximately 30% to 40% of all chiral-controlled internucleotide bonds in the oligonucleotide are Rp. In some embodiments, approximately 35% to 40% of all chiral-controlled internucleotide bonds in the oligonucleotide are Rp.

[0318] In some embodiments, the natural phosphate bond is optionally modified, for example, sugar-modified (e.g., R as described herein) instead of the Rp internucleotide bond. 5s It can be used similarly with 5'-modifications such as those mentioned above. In some embodiments, the modification improves the stability of the native phosphate bond.

[0319] In some embodiments, the disclosure provides oligonucleotides having a skeletal chiral center pattern as described herein. In some embodiments, oligonucleotides in a chiral-controlled oligonucleotide composition share a common skeletal chiral center pattern as described herein.

[0320] In some embodiments, at least about 25% of the internucleotide bonds of the MAPT oligonucleotide are chiral-controlled and have Sp-bound phosphorus. In some embodiments, at least about 30% of the internucleotide bonds of the oligonucleotide are chiral-controlled and have Sp-bound phosphorus. In some embodiments, at least about 40% of the internucleotide bonds of the provided oligonucleotide are chiral-controlled and have Sp-bound phosphorus. In some embodiments, at least about 50% of the internucleotide bonds of the provided oligonucleotide are chiral-controlled and have Sp-bound phosphorus. In some embodiments, at least about 60% of the internucleotide bonds of the provided oligonucleotide are chiral-controlled and have Sp-bound phosphorus. In some embodiments, at least about 65% of the internucleotide bonds of the provided oligonucleotide are chiral-controlled and have Sp-bound phosphorus. In some embodiments, at least about 70% of the internucleotide bonds of the provided oligonucleotide are chiral-controlled and have Sp-bound phosphorus. In some embodiments, at least about 75% of the internucleotide bonds of the provided oligonucleotide are chiral-controlled and have Sp-bound phosphorus. In some embodiments, at least about 80% of the internucleotide bonds of the provided oligonucleotide are chiral-controlled and have Sp-bound phosphorus. In some embodiments, at least about 85% of the internucleotide bonds of the provided oligonucleotide are chiral-controlled and have Sp-linked phosphorus. In some embodiments, at least about 90% of the internucleotide bonds of the provided oligonucleotide are chiral-controlled and have Sp-linked phosphorus. In some embodiments, at least about 95% of the internucleotide bonds of the provided oligonucleotide are chiral It is controlled and contains Sp-bound phosphorus.

[0321] In some embodiments, the Disclosure provides chiral-controlled oligonucleotide compositions, for example, chiral-controlled MAPT oligonucleotide compositions, wherein the composition comprises a plurality of oligonucleotides at non-random or controlled levels, wherein the plurality of oligonucleotides share a common base sequence and independently share the same binding phosphorus configuration at 1-50, 1-40, 1-30, 1-25, 1-20, 1-15, 1-10, 5-50, 5-40, 5-30, 5-25, 5-20, 5-15, 5-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 or more chiral internucleotide bonds.

[0322] In some embodiments, the MAPT oligonucleotide contains 2 to 30 chiral-controlled internucleotide bonds. In some embodiments, the provided oligonucleotide composition contains 5 to 30 chiral-controlled internucleotide bonds. In some embodiments, the provided oligonucleotide composition contains 10 to 30 chiral-controlled internucleotide bonds.

[0323] In some embodiments, the percentage is about 5% to 100%. In some embodiments, the percentage is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 965%, 96%, 98%, or 99%. In some embodiments, the percentage is about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 965%, 96%, 98%, or 99%.

[0324] In some embodiments, the chiral center pattern of the skeleton in MAPT oligonucleotides is i o -i s -i o -i s -i o i o-i s -i s -i s -i o 、i o -i s -i s -i s -i o -i s 、i s -i o -i s -i o 、i s -i o -i s -i o 、i s -i o -i s -i o -i s 、i s -i o -i s -i o -i s -i o 、i s -i o -i s -i o -i s -i o -i s -i o 、i s -i o -i s -i s -i s -i o 、i s -i s -i o -i s -i s -i s -i o -i s -i s 、i s -i s -i s -i o -i s -i o -i s -i s -i s 、i s -i s -i s -i s -i o -is -i o -i s -i s -i s -i s i s -i s -i s -i s -i s i s -i s -i s -i s -i s -i s i s -i s -i s -i s -i s -i s -i s i s -i s -i s -i s -i s -i s -i s -i s i s -i s -i s -i s -i s -i s -i s -i s -i s , or i r -i r -i r The pattern (where i s represents an internucleotide bond in the Sp configuration; i o represents an achiral internucleotide bond; and i r This includes an internucleotide bond in the Rp configuration.

[0325] In some embodiments, the Sp-configured (having Sp-bound phosphorus) internucleotide bond is a phosphorothioate internucleotide bond. In some embodiments, the achiral internucleotide bond is a native phosphate bond. In some embodiments, the Rp-configured (having Rp-bound phosphorus) internucleotide bond is a phosphorothioate internucleotide bond. In some embodiments, each Sp-configured internucleotide bond is a phosphorothioate internucleotide bond. In some embodiments, each achiral internucleotide bond is a native phosphate bond. In some embodiments, each Rp-configured internucleotide bond is a phosphorothioate internucleotide bond. In some embodiments, each Sp-configured internucleotide bond is a phosphorothioate internucleotide bond, each achiral internucleotide bond is a native phosphate bond, and each Rp-configured internucleotide bond is a phosphorothioate internucleotide bond.

[0326] In some embodiments, the skeletal chiral center pattern (e.g., in an oligonucleotide, e.g., in a MAPT oligonucleotide or in an oligonucleotide, e.g., in the core or wings of a MAPT oligonucleotide or in two wings) is OpSpOpSpOp, OpSpSpSpOp, OpSpSpSpOpSp, SpOpSpOp, SpOpSpOp, SpOpSpOpSpSp, SpOpSpOpSpOpSpOp, SpOpSpSpSpOpSpOpSp, SpSpSpOpSpOpSpSp The patterns include SpSpSpSpOpSpOpSpSpSpSp, SpSpSpSpSp, SpSpSpSpSpSp, SpSpSpSpSpSpSp, SpSpSpSpSpSpSpSp, SpSpSpSpSpSpSpSpSp, or RpRpRp (where each Rp and Sp is independently a chiral-controlled internucleotide bond binding phosphorus arrangement (in some embodiments, each Rp and Sp is independently a chiral-controlled phosphorothioate internucleotide bond binding phosphorus arrangement), and each Op is independently an achiral binding phosphorus in the native phosphate bond).

[0327] In some embodiments, the skeletal chiral center pattern (e.g., an oligonucleotide, e.g., a MAPT oligonucleotide, or a portion thereof) is or includes the skeletal chiral center patterns listed in Table 1A.

[0328] In some embodiments, an internucleotide bond attached to a wing nucleoside and a core nucleoside is considered one of the core internucleotide bonds, for example, when describing the type, modification, number, and / or pattern of core internucleotide bonds. In some embodiments, each internucleotide bond attached to a wing nucleoside and a core nucleoside is considered one of the core internucleotide bonds, for example, when describing the type, modification, number, and / or pattern of core internucleotide bonds. In some embodiments, a core internucleotide bond is attached to two core nucleosides. In some embodiments, a core internucleotide bond is attached to a core nucleoside and a wing nucleoside. In some embodiments, each core internucleotide bond is independently attached to two core nucleosides, or to a core nucleoside and a wing nucleoside. In some embodiments, each wing internucleotide bond is independently attached to two wing nucleosides.

[0329] In some embodiments, each MAPT oligonucleotide in a chiral-controlled oligonucleotide composition contains a different type of internucleotide bond. In some embodiments, a MAPT oligonucleotide contains at least one native phosphate bond and at least one modified internucleotide bond. In some embodiments, a MAPT oligonucleotide contains at least one native phosphate bond and at least two modified internucleotide bonds. In some embodiments, a MAPT oligonucleotide contains at least one native phosphate bond and at least three modified internucleotide bonds. In some embodiments, a MAPT oligonucleotide contains at least one native phosphate bond and at least four modified internucleotide bonds. In some embodiments, a MAPT oligonucleotide contains at least one native phosphate bond and at least five modified internucleotide bonds. In some embodiments, a MAPT oligonucleotide contains at least one native phosphate bond and 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 modified internucleotide bonds. In some embodiments, the modified internucleotide bond is a phosphorothioate internucleotide bond. In some embodiments, each modified internucleotide The phosphate bond is a phosphorothioate internucleotide bond. In some embodiments, the modified internucleotide bond is a phosphorothioate triester internucleotide bond. In some embodiments, each modified internucleotide bond is a phosphorothioate triester internucleotide bond. In some embodiments, the MAPT oligonucleotide comprises at least one native phosphate bond and at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 consecutive modified internucleotide bonds. In some embodiments, the MAPT oligonucleotide comprises at least one native phosphate bond and at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 consecutive phosphorothioate internucleotide bonds. In some embodiments, the MAPT oligonucleotide comprises at least one native phosphate bond and at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 consecutive phosphorothioate triester internucleotide bonds.

[0330] In some embodiments, the oligonucleotides in the chiral-controlled oligonucleotide composition each comprise at least two internucleotide bonds having different stereochemistry and / or different phosphorus modifications. In some embodiments, the at least two internucleotide bonds have different stereochemistry, and each of these oligonucleotides comprises a skeletal chiral center pattern containing alternating linked phosphorus stereochemistry.

[0331] In some embodiments, the bond includes a chiral auxiliary, which is used, for example, to control the stereoselectivity of reactions in the oligonucleotide synthesis cycle, such as coupling reactions. In some embodiments, the phosphorothioate triester bond does not include a chiral auxiliary. In some embodiments, the phosphorothioate triester bond is intentionally maintained until the oligonucleotide composition is administered to the target and / or during administration.

[0332] In some embodiments, all chiral centers in the oligonucleotide except the chiral-binding phosphorus center are sterically defined (e.g., carbon chiral centers in sugars, which are defined, for example, in phosphoramidites for oligonucleotide synthesis). The purity, in particular stereochemical purity, and especially diastereomer purity, of many oligonucleotides and their compositions can be controlled by the stereoselectivity at the chiral-binding phosphorus (diastereoselectivity in many oligonucleotide synthesis, as those skilled in the art will understand, in oligonucleotides containing two or more chiral centers) in the coupling step when forming the chiral internucleotide bond. In some embodiments, the coupling step has 60% stereoselectivity at the binding phosphorus (diastereoselectivity if other chiral centers are present). The novel internucleotide bond formed after such a coupling step can be said to have 60% stereochemical purity (for oligonucleotides, typically diastereomer purity considering the presence of other chiral centers). In some embodiments, each coupling step independently has at least 60% stereoselectivity. In some embodiments, each coupling step independently has at least 70% stereoselectivity. In some embodiments, each coupling step independently has at least 80% stereoselectivity. In some embodiments, each coupling step independently has at least 85% stereoselectivity. In some embodiments, each coupling step independently has at least 90% stereoselectivity. In some embodiments, each coupling step independently has at least 91% stereoselectivity. In some embodiments, each coupling step independently has at least 92% stereoselectivity. In some embodiments, each coupling step independently has at least It has 93% stereoselectivity. In some embodiments, each coupling step independently has at least 94% stereoselectivity. In some embodiments, each coupling step independently has at least 95% stereoselectivity. In some embodiments, each coupling step independently has at least 96% stereoselectivity. In some embodiments, each coupling step independently has at least 97% stereoselectivity. In some embodiments, each coupling step independently has at least 98% stereoselectivity. In some embodiments, each coupling step independently has at least 99% stereoselectivity. In some embodiments, each coupling step independently has at least 99.5% stereoselectivity. In some embodiments, each coupling step independently has virtually 100% stereoselectivity. In some embodiments, the coupling steps have virtually 100% stereoselectivity in that each detectable product from the coupling steps, as analyzed by analytical methods (e.g., NMR, HPLC, etc.), has the intended stereoselectivity. In some embodiments, chiral-controlled internucleotide bonds are typically formed with stereoselectivity of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99.5%, or virtually 100% (at least 90% in some embodiments; at least 95% in some embodiments; at least 96% in some embodiments; at least 97% in some embodiments; at least 98% in some embodiments; and at least 99% in some embodiments).In some embodiments, the chiral-controlled internucleotide bond has a stereochemical purity of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99.5%, or virtually 100% (at least 90% in some embodiments; at least 95% in some embodiments; at least 96% in some embodiments; at least 97% in some embodiments; at least 98% in some embodiments; and at least 99% in some embodiments) of the chiral-bound phosphorus (typically diastereomer purity for oligonucleotides having multiple chiral centers). In some embodiments, each chiral-controlled internucleotide bond independently has a stereochemical purity of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99.5%, or virtually 100% (at least 90% in some embodiments; at least 95% in some embodiments; at least 96% in some embodiments; at least 97% in some embodiments; at least 98% in some embodiments; and at least 99% in some embodiments) to its chiral-bound phosphorus (typically diastereomer purity for oligonucleotides having multiple chiral centers). In some embodiments, non-chiral-controlled internucleotide bonds are typically formed with stereoselectivity of 60%, 70%, 80%, 85%, or less than 90% (less than 60% in some embodiments; less than 70% in some embodiments; less than 80% in some embodiments; less than 85% in some embodiments; and less than 90% in some embodiments). In some embodiments, each chiral-uncontrolled internucleotide bond is formed independently with stereoselectivity of 60%, 70%, 80%, 85%, or less than 90% (less than 60% in some embodiments; less than 70% in some embodiments; less than 80% in some embodiments; less than 85% in some embodiments; less than 90% in some embodiments).In some embodiments, the chiral-uncontrolled internucleotide bond has a stereochemical purity of less than 60%, 70%, 80%, 85%, or 90% (in some embodiments, less than 60%; in some embodiments, less than 70%; in some embodiments, less than 80%; in some embodiments, less than 85%; in some embodiments, less than 90%) of its chiral-bound phosphorus (typically diastereomer purity for oligonucleotides having multiple chiral centers). In some embodiments, each chiral-uncontrolled internucleotide bond independently has a stereochemical purity of less than 60%, 70%, 80%, 85%, or 90% (in some embodiments, less than 60%; in some embodiments, less than 70%; in some embodiments, less than 90%) of its chiral-bound phosphorus. In its state, it has a stereochemical purity of less than 80%; in some embodiments, less than 85%; and in some embodiments, less than 90% (typically diastereomer purity for oligonucleotides having multiple chiral centers).

[0333] In some embodiments, at least one, two, three, four, five, six, seven, eight, nine, or ten couplings of a monomer (phosphoamidite for oligonucleotide synthesis in many embodiments, as those skilled in the art will understand) independently have stereoselectivity of about 60%, 70%, 80%, 85%, or less than 90% [for oligonucleotide synthesis, typically diastereoselectivity with respect to one or more binding line chiral centers formed]. In some embodiments, at least one coupling has stereoselectivity of about 60%, 70%, 80%, 85%, or less than 90%. In some embodiments, at least two couplings independently have stereoselectivity of about 60%, 70%, 80%, 85%, or less than 90%. In some embodiments, at least three couplings independently have stereoselectivity of about 60%, 70%, 80%, 85%, or less than 90%. In some embodiments, at least four couplings independently have stereoselectivity of about 60%, 70%, 80%, 85%, or less than 90%. In some embodiments, at least five couplings independently have stereoselectivity of about 60%, 70%, 80%, 85%, or less than 90%. In some embodiments, each coupling independently has stereoselectivity of about 60%, 70%, 80%, 85%, or less than 90%. In some embodiments, each uncontrolled chiral internucleotide bond is formed with stereoselectivity of about 60%, 70%, 80%, 85%, or less than 90%. In some embodiments, the stereoselectivity is less than about 60%. In some embodiments, the stereoselectivity is less than about 70%. In some embodiments, the stereoselectivity is less than about 80%. In some embodiments, the stereoselectivity is less than about 90%. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 couplings independently have stereoselectivity of less than about 90%. In some embodiments, at least 1 coupling has stereoselectivity of less than about 90%. In some embodiments, at least 2 couplings have stereoselectivity of less than about 90%.In some embodiments, at least three couplings have stereoselectivity of less than about 90%. In some embodiments, at least four couplings have stereoselectivity of less than about 90%. In some embodiments, at least five couplings have stereoselectivity of less than about 90%. In some embodiments, each coupling independently has stereoselectivity of less than about 90%. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 couplings independently have stereoselectivity of less than about 85%. In some embodiments, each coupling independently has stereoselectivity of less than about 85%. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 couplings independently have stereoselectivity of less than about 80%. In some embodiments, each coupling independently has stereoselectivity of less than about 80%. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 couplings independently have stereoselectivity of less than about 70%. In some embodiments, each coupling independently has stereoselectivity of less than about 70%.

[0334] In some embodiments, the oligonucleotides and compositions of this disclosure have high purity. In some embodiments, the oligonucleotides and compositions of this disclosure have high stereochemical purity. In some embodiments, the stereochemical purity, for example, the diastereomer purity, is about 60% to 100%. In some embodiments, the diastereomer purity is It is approximately 60% to 100%. In some embodiments, the percentage is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 93%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the percentage is at least 80%, 85%, 90%, 91%, 92%, 93%, 93%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the percentage is at least 90%, 91%, 92%, 93%, 93%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the diastereomer purity is at least 60%. In some embodiments, the diastereomer purity is at least 70%. In some embodiments, the diastereomer purity is at least 80%. In some embodiments, the diastereomer purity is at least 85%. In some embodiments, the diastereomer purity is at least 90%. In some embodiments, the diastereomer purity is at least 91%. In some embodiments, the diastereomer purity is at least 92%. In some embodiments, the diastereomer purity is at least 93%. In some embodiments, the diastereomer purity is at least 94%. In some embodiments, the diastereomer purity is at least 95%. In some embodiments, the diastereomer purity is at least 96%. In some embodiments, the diastereomer purity is at least 97%. In some embodiments, the diastereomer purity is at least 98%. In some embodiments, the diastereomer purity is at least 99%. In some embodiments, the diastereomer purity is at least 99.5%.

[0335] In some embodiments, the compounds of the Disclosure (e.g., oligonucleotides, chiral auxiliary groups, etc.) contain multiple chiral elements (e.g., multiple carbon and / or phosphorus (e.g., binding phosphorus of chiral internucleotide bonds) chiral centers). In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or more chiral elements of the provided compound (e.g., oligonucleotides) each independently have the diastereomer purity as described herein. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or more chiral carbon centers of the provided compound each independently have the diastereomer purity as described herein. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or more chiral phosphorus centers of the provided compound each independently have the diastereomer purity as described herein. In some embodiments, each chiral element independently has the diastereomer purity as described herein. In some embodiments, each chiral center independently has the diastereomer purity as described herein. In some embodiments, each chiral carbon center independently has the diastereomer purity as described herein. In some embodiments, each chiral phosphorus center independently has the diastereomer purity as described herein. In some embodiments, each chiral phosphorus center independently has a diastereomer purity of at least 90%, 91%, 92%, 93%, 93%, 95%, 96%, 97%, 98%, or 99% or more.

[0336] As those skilled in the art will understand, in some embodiments, the diastereoselectivity of a coupling or the diastereomer purity of a chiral-linked phosphorus center can be evaluated by the diastereoselectivity of dimer formation or the diastereomer purity of a dimer prepared under the same or equivalent conditions, where the dimer has the same 5'- and 3'-nucleosides and internucleotide bonds.

[0337] Identification or confirmation of the stereochemistry of chiral elements (e.g., arrangement of chiral-bonded phosphorus) and / or the chiral center pattern of the skeleton, and / or stereoselectivity (e.g., diastereoselectivity of the coupling step in oligonucleotide synthesis) and / or stereochemical purity (e.g., i Various techniques can be used to evaluate the internucleotide bond and diastereomer purity of compounds (e.g., oligonucleotides). Specific techniques include NMR [e.g., 1D (one-dimensional) and / or 2D (two-dimensional)]. 1 H- 31 Methods such as P HETCOR (heteronuclear correlation spectroscopy), HPLC, RP-HPLC, mass spectrometry, LC-MS, and cleavage of internucleotide bonds by stereospecific nucleases can be used individually or in combination. Useful exemplary nucleases include benzoases, micrococcus nucleases, and svPDE (snake venom phosphodiesterases) that are specific to certain internucleotide bonds with Rp-binding phosphorus (e.g., Rp phosphorothioate bonds); and nucleases P1, mangu bean nucleases, and S1 that are specific to internucleotide bonds with Sp-binding phosphorus (e.g., Sp phosphorothioate bonds). While we do not wish to be bound by any particular theory, this disclosure notes that, at least in some cases, cleavage of oligonucleotides by certain nucleases may be influenced by structural elements, such as chemical modifications (e.g., 2'-modification of sugars), nucleotide sequence, or stereochemical context. For example, in some cases, benzoases and micrococcus nucleases specific to internucleotide bonds containing Rp-binding phosphorus were observed to be unable to cleave isolated Rp-phosphorothioate internucleotide bonds where the Sp-phosphorothioate internucleotide bond was located laterally.

[0338] In some embodiments, oligonucleotides sharing a common base sequence, a common skeletal bonding pattern, and a common skeletal chiral center pattern share a common skeletal phosphorus modification pattern and a common base modification pattern. In some embodiments, oligonucleotide compositions sharing a common base sequence, a common skeletal bonding pattern, and a common skeletal chiral center pattern share a common skeletal phosphorus modification pattern and a common nucleoside modification pattern. In some embodiments, oligonucleotides sharing a common base sequence, a common skeletal bonding pattern, and a common skeletal chiral center pattern have the same structure.

[0339] In some embodiments, the Disclosure provides an oligonucleotide composition comprising a plurality of oligonucleotides having the ability to induce MAPT knockdown, wherein the oligonucleotides are of a specific oligonucleotide type, and the composition is chiral-controlled in that the oligonucleotides of the specific oligonucleotide type are highly purified compared to a substantially racemic formulation of oligonucleotides having the same base sequence.

[0340] In some embodiments, oligonucleotides having a common base sequence, a common skeletal bonding pattern, and a common skeletal chiral center pattern have a common skeletal phosphorus modification pattern and a common base modification pattern. In some embodiments, oligonucleotides having a common base sequence, a common skeletal bonding pattern, and a common skeletal chiral center pattern have a common skeletal phosphorus modification pattern and a common nucleoside modification pattern. In some embodiments, oligonucleotides having a common base sequence, a common skeletal bonding pattern, and a common skeletal chiral center pattern have the same structure.

[0341] In some embodiments, the Disclosure provides MAPT oligonucleotide compositions comprising a plurality of oligonucleotides. In some embodiments, the Disclosure provides chiral-controlled oligonucleotide compositions of MAPT oligonucleotides. In some embodiments, the Disclosure provides MAPT oligonucleotides which are or are complementary to the MAPT sequences disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa). In some embodiments, the Disclosure provides MAPT oligonucleotides which are or are complementary to the MAPT sequences disclosed herein (e.g., various nucleotide sequences in Table 1). The disclosure provides MAPT oligonucleotides having nucleotide sequences containing a sequence. In some embodiments, the disclosure provides MAPT oligonucleotides having a sequence containing 15 adjacent nucleotides of a MAPT sequence or a portion thereof disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa) or a nucleotide sequence complementary thereto. In some embodiments, the disclosure provides MAPT oligonucleotides having a sequence containing 15 adjacent nucleotides having 0 to 3 mismatches of a MAPT sequence or a portion thereof disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa) or a nucleotide sequence complementary thereto. In some embodiments, the disclosure provides MAPT oligonucleotide compositions in which the MAPT oligonucleotide contains at least one chiral internucleotide bond that is not chiral controlled. In some embodiments, the Disclosure provides MAPT oligonucleotides comprising an uncontrolled chiral internucleotide bond, wherein the nucleotide sequence of the MAPT oligonucleotide is either a MAPT sequence or a portion thereof as disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa) or a nucleotide sequence complementary thereto. In some embodiments, the Disclosure provides MAPT oligonucleotide compositions comprising an uncontrolled chiral internucleotide bond, wherein the nucleotide sequence of the MAPT oligonucleotide is either a MAPT sequence or a portion thereof as disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa) or a nucleotide sequence complementary thereto.In some embodiments, the disclosure provides MAPT oligonucleotides comprising an uncontrolled chiral internucleotide bond, wherein the nucleotide sequence of the MAPT oligonucleotide comprises 15 adjacent nucleotides of a MAPT sequence or a portion thereof disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa) or a nucleotide sequence complementary thereto. In some embodiments, the Disclosure provides MAPT oligonucleotides comprising chiral-controlled chiral internucleotide links, wherein the nucleotide sequence of the MAPT oligonucleotide comprises a MAPT sequence or a portion thereof as disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa) or a nucleotide sequence complementary thereto. In some embodiments, the Disclosure provides MAPT oligonucleotide compositions comprising chiral-controlled chiral internucleotide links, wherein the nucleotide sequence of the MAPT oligonucleotide comprises a MAPT sequence or a portion thereof as disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa) or a nucleotide sequence complementary thereto. In some embodiments, the Disclosure provides MAPT oligonucleotides comprising chiral-controlled chiral internucleotide links, wherein the nucleotide sequence of the MAPT oligonucleotide comprises 15 adjacent nucleotides of a MAPT sequence or a portion thereof disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa) or a nucleotide sequence complementary thereto.In some embodiments, the Disclosure provides MAPT oligonucleotides comprising chiral-controlled chiral internucleotide bonds, wherein the nucleotide sequence of the MAPT oligonucleotide comprises 15 adjacent nucleotides having 0 to 3 mismatches of MAPT sequences or portions thereof disclosed herein (e.g., various nucleotide sequences in Table 1, where each T can be independently replaced by U, and vice versa) or a complementary nucleotide sequence.

[0342] In some embodiments, oligonucleotides of the same oligonucleotide type have a common skeletal phosphate modification pattern and a common nucleoside modification pattern. In some embodiments, oligonucleotides of the same oligonucleotide type have a common sugar modification pattern. In some embodiments, oligonucleotides of the same oligonucleotide type have a common base modification pattern. In some embodiments, oligonucleotides of the same oligonucleotide type have a common nucleoside modification pattern. In some embodiments, oligonucleotides of the same oligonucleotide type have the same chemical composition. In many embodiments, oligonucleotides of the same oligonucleotide type are identical. In some embodiments, oligonucleotides of the same oligonucleotide type are of the same oligonucleotide (as those skilled in the art will understand, such oligonucleotides may exist independently as one of various forms of the oligonucleotide and may be the same or different forms of the oligonucleotide). In some embodiments, oligonucleotides of the same oligonucleotide type are independently of the same oligonucleotide or a pharmaceutically acceptable salt thereof.

[0343] In some embodiments, multiple oligonucleotides or specific oligonucleotide types in the provided oligonucleotide composition are MAPT oligonucleotides. In some embodiments, the disclosure provides a chiral-controlled MAPT oligonucleotide composition comprising multiple MAPT oligonucleotides, wherein these oligonucleotides are 1) Common base sequence; 2) Common skeletal connection patterns; and 3) The same phosphorus stereochemistry in one or more chiral internucleotide bonds (chiral-controlled internucleotide bonds) Share, The composition features higher purity for some of the oligonucleotides compared to a substantially racemic formulation of oligonucleotides that share a common base sequence and skeletal bonding pattern.

[0344] In some embodiments, as used herein, "one or more" or "at least one" means 1 to 50, 1 to 40, 1 to 30, 1 to 25, 1 to 20, 1 to 15, 1 to 10, 5 to 50, 5 to 40, 5 to 30, 5 to 25, 5 to 20, 5 to 15, 5 to 10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more.

[0345] In some embodiments, the oligonucleotide type is further defined by an additional chemical part, if present.

[0346] In some embodiments, the percentage is at least about 10%. In some embodiments, the percentage is at least about 20%. In some embodiments, the percentage is at least about 30%. In some embodiments, the percentage is at least about 40%. In some embodiments, the percentage is at least about 50%. In some embodiments, the percentage is at least about 60%. In some embodiments, the percentage is at least about 70%. In some embodiments, the percentage is at least about 75%. In some embodiments, the percentage is at least about 80%. In some embodiments, the percentage is at least about 85%. In some embodiments, the percentage is at least about 90%. In some embodiments, the percentage is at least about 91%. In some embodiments, the percentage is at least about 92%. Yes. In some embodiments, the percentage is at least about 93%. In some embodiments, the percentage is at least about 94%. In some embodiments, the percentage is at least about 95%. In some embodiments, the percentage is at least about 96%. In some embodiments, the percentage is at least about 97%. In some embodiments, the percentage is at least about 98%. In some embodiments, the percentage is at least about 99%. In some embodiments, the percentage is (DS) nc (wherein DS and nc are, independently, as described herein) or greater than.

[0347] In some embodiments, multiple oligonucleotides, for example, MAPT oligonucleotides, share the same chemical structure. In some embodiments, multiple oligonucleotides, for example, MAPT oligonucleotides, are identical (the same stereoisomer). In some embodiments, a chiral-controlled oligonucleotide composition, for example, a chiral-controlled MAPT oligonucleotide composition, is a sterically pure oligonucleotide composition in which multiple oligonucleotides are identical (the same stereoisomer), and this composition does not contain any other stereoisomers. Those skilled in the art will understand that processing, selection, purification, etc., may not achieve perfection, and therefore one or more other stereoisomers may be present as impurities.

[0348] In some embodiments, the provided composition is characterized by a decrease in the level of the target nucleic acid and / or the product encoded therein when it comes into contact with the target nucleic acid [e.g., MAPT transcript (e.g., pre-mRNA, mature mRNA, other types of RNA, etc. that hybridize with the oligonucleotide of the composition)] compared to that observed under reference conditions. In some embodiments, the reference conditions are selected from the group consisting of the absence of the composition, the presence of a reference composition, and combinations thereof. In some embodiments, the reference condition is the absence of the composition. In some embodiments, the reference condition is the presence of a reference composition. In some embodiments, the reference composition is a composition in which the oligonucleotide does not hybridize with the target nucleic acid. In some embodiments, the reference composition is a composition in which the oligonucleotide does not contain a sequence sufficiently complementary to the target nucleic acid. In some embodiments, the provided composition is a chiral-controlled oligonucleotide composition, and the reference composition is a non-chiral-controlled oligonucleotide composition, which is otherwise identical but not chiral-controlled (e.g., a racemic formulation of oligonucleotides having the same chemical composition as the oligonucleotide(s) in the chiral-controlled oligonucleotide composition).

[0349] In some embodiments, the present disclosure provides a chiral-controlled MAPT oligonucleotide composition comprising a plurality of MAPT oligonucleotides having the ability to induce MAPT knockdown, whe...

Claims

1. An oligonucleotide wherein the base sequence of the oligonucleotide includes at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 adjacent bases that are identical to or complementary to the base sequence of the MAPT gene or its transcript, and the oligonucleotide includes one or more modified sugars, one or more modified nucleic acid bases, and / or one or more modified internucleotide bonds.

2. The oligonucleotide according to claim 1, wherein the base sequence of the oligonucleotide includes at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 adjacent bases of a base sequence complementary to the MAPT transcript.

3. The oligonucleotide according to claim 1, wherein the base sequence of the oligonucleotide includes at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 adjacent bases of a base sequence complementary to MAPT mRNA.

4. The oligonucleotide according to claim 1, wherein the base sequence of the oligonucleotide includes at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 adjacent bases of a base sequence complementary to the intron 11 of MAPT RNA.

5. The base sequence of the aforementioned PLC is ACGTTGCAGTGTTCCACUAU, ACTATCCTCTCTCCAGCUCCU, AGTGTTCCAACTATCCCUUCCUU, ATCCTCTCTCCAGCCUGCCA, CACGTTGCAGTGTTCCAACCUA, CACTATCCTCTCTCCAGCCU CC, CAGTGTTCCCACTATCCUCCU, CCACGTTGCAGTGTTCCACU, CCACTATCCTCCTTCAGCUC, CGTT GCAGTGTTCCACUAUC, CTATCCTCCTTCAGCUCCUG, GCAGTGTTCCACTATCCUCC, GTGTTCCACTAT The iSCSI according to any one of claims 1 to 4, comprising at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 adjacent bases of CCTCCUUC, GTTCCACATCCTCCUUCAG, GUGUUCCACATCCTCTCTCTC, TATCCTCTCTCCAGCTCCUGC, TCCACATCCTCTCTCCAGC, TGCAGTGTTCCCACTAUCCUC, TGTTCCCATCCTCCUUCA, TTCCACATCCTCTCTUCAGC, or TTGCAGTGTTCCCACTAUCCCU (wherein each T can be independently substituted with U, and vice versa).

6. An oligonucleotide according to any one of claims 1 to 5, comprising about 20 nucleic acid bases independently selected from A, T, C, G, U and their tautomers, which are optionally substituted.

7. The oligonucleotide according to any one of claims 1 to 6, comprising a 5'-wing-core-wing-3' portion, wherein the 5'-wing and the 3'-wing each independently contain two or more modified sugars.

8. The aforementioned 5'-wing is 2'-OR (wherein R is optionally substituted C). １～４ The oligonucleotide according to claim 7, comprising two or more sugars, each independently containing an aliphatic sugar.

9. The oligonucleotide according to claim 8, wherein the 5'-wing comprises one or more 2'-MOE modified sugars.

10. The oligonucleotide according to any one of claims 7 to 9, wherein each sugar in the 5'-wing includes a 2'-modified sugar.

11. Each sugar in the 5'-wing comprises the same modification as the oligonucleotide according to any one of claims 7 to 10.

12. The oligonucleotide according to any one of claims 7 to 11, wherein the 5'-wing comprises a 4, 5, or 7 nucleoside.

13. The oligonucleotide according to any one of claims 7 to 12, wherein the 5'-wing comprises one or more phosphorothioate internucleotide bonds.

14. The oligonucleotide according to any one of claims 7 to 13, wherein the 5'-wing comprises one or more non-negatively charged internucleotide bonds.

15. The oligonucleotide according to any one of claims 7 to 14, wherein the 5'-wing comprises one or more natural phosphate bonds.

16. The aforementioned 3'-wing is 2'-OR (wherein R is optionally replaced by C). １～４ An oligonucleotide according to any one of claims 7 to 15, comprising two or more sugars, each independently containing an aliphatic sugar.

17. The oligonucleotide according to any one of claims 7 to 16, wherein the sugar modification pattern of the 5'-wing is different from that of the 3'-wing.

18. The oligonucleotide according to any one of claims 7 to 17, wherein the 3'-wing includes a sugar modification not present in the 5'-wing.

19. The oligonucleotide according to any one of claims 7 to 18, wherein the 3'-wing comprises one or more 2'-OMe-modified sugars.

20. The oligonucleotide according to any one of claims 7 to 19, wherein each sugar in the 3'-wing includes a 2'-modified sugar.

21. Each sugar in the 3'-wing comprises the same modification as the oligonucleotide according to any one of claims 7 to 20.

22. The oligonucleotide according to any one of claims 7 to 21, wherein the 3'-wing comprises a 4, 5, or 7 nucleoside.

23. The oligonucleotide according to any one of claims 7 to 22, wherein the 3'-wing comprises one or more phosphorothioate internucleotide bonds.

24. The oligonucleotide according to any one of claims 7 to 23, wherein the 3'-wing comprises one or more non-negatively charged internucleotide bonds.

25. The oligonucleotide according to any one of claims 7 to 24, wherein the 3'-wing comprises one or more natural phosphate bonds.

26. The oligonucleotide according to any one of claims 7 to 25, wherein each phosphorothioate internucleotide bond in the 5'-wing is Sp.

27. The oligonucleotide according to any one of claims 7 to 26, wherein each phosphorothioate internucleotide bond in the 3'-wing is Sp.

28. The oligonucleotide according to any one of claims 7 to 27, wherein each non-negatively charged internucleotide bond in the 5'-wing is n001.

29. The oligonucleotide according to any one of claims 7 to 28, wherein each non-negatively charged internucleotide bond in the 3'-wing is n001.

30. The oligonucleotide according to claim 28 or 29, wherein n001 is Rp.

31. The aforementioned core is 2'-OR sugar modified (wherein R is optionally substituted C). １～４ An oligonucleotide according to any one of claims 7 to 30, which does not contain (aliphatic).

32. The oligonucleotide according to any one of claims 7 to 31, wherein each sugar in the core is a natural DNA sugar (containing two 2'-H atoms).

33. The oligonucleotide according to any one of claims 7 to 32, wherein the core comprises about 7, 8, 9, 10, or 11 nucleosides.

34. The oligonucleotide according to any one of claims 7 to 33, wherein each internucleotide bond bound to the nucleoside in the core is independently a phosphorothioate internucleotide bond.

35. The oligonucleotide according to any one of claims 1 to 34, wherein the chiral center pattern of the oligonucleotide skeleton is [(Rp)n(Sp)m]y (wherein each of n, m, and y is independently 1 to 25) or includes the same.

36. The oligonucleotide according to any one of claims 1 to 35, wherein the chiral center pattern of the oligonucleotide skeleton is (Sp)t[(Rp)n(Sp)m]y (wherein each of t, n, m and y is independently 1 to 25).

37. The oligonucleotide according to any one of claims 7 to 36, wherein the chiral center pattern of the core skeleton is [(Rp)n(Sp)m]y (wherein each of n, m, and y is independently 1 to 25).

38. The oligonucleotide according to any one of claims 7 to 37, wherein the chiral center pattern of the core skeleton is (Sp)t[(Rp)n(Sp)m]y (wherein each of t, n, m and y is independently 1 to 25).

39. The oligonucleotide according to any one of claims 35 to 38, wherein at least one n is 1.

40. The oligonucleotide according to any one of claims 35 to 39, wherein each n is 1.

41. The oligonucleotide according to any one of claims 35 to 40, wherein each m is independently 2 to 25.

42. The oligonucleotide according to any one of claims 35 to 41, wherein y is 1.

43. The oligonucleotide according to any one of claims 35 to 42, wherein y is 2 to 5.

44. The oligonucleotide according to any one of claims 1 to 43, wherein the base sequence of the oligonucleotide is approximately 90% to 100% complementary to the base sequence in the MAPT transcript.

45. The base sequence of PLC is ACGTTGCAGTGTTCCACUAU, ACTATCCTCTCTCCAGCUCCU, AGTGTTCCAACTATCCCUUCCUU, ATCCTCTCTCCAGCTCUGCA, CACGTTGCAGTGTTCCACUA, CACTATCCTCTCTCCAGCUCC, CAGTGTTCCAACTATCCCUUCCU, CCACGTTGCAGTGTTCCACU, CCACTATCCTCTCTCCAGCUC, CGTTTGCAGTGTTCCACUAUUC, CTATCCTCTCTCCAGCUCUCUG, GCAGTGTTCCA The CiNii relation according to any one of claims 1 to 44, wherein the relation is CTATCCUCC, GTGTTCCACACTATCCTCCUUC, GTTCCACACTATCCTCCUUCAG, GUGUUCCACTATCCTCTCTCTC, TATCCTCTCTCCAGCTCCUGC, TCCACTATCCTCTCTCCAGCCU, TGCAGTGTTCCCACTAUCCUC, TGTTCCCACACTATCCTCCUUCA, TTCCACACTATCCTCTCTUCAGC, or TTGCAGTGTTCCCACTAUCCCU (wherein each T can be independently substituted with U, and vice versa).

46. The oligonucleotide according to any one of claims 1 to 45, wherein the base sequence of the oligonucleotide is GTGTTCCACTATCCTCCUUC (wherein each T may be independently substituted with U, and vice versa).

47. The oligonucleotide according to any one of claims 1 to 46, wherein the base sequence of the oligonucleotide is GTGTTCCACTATCCTCCUUC.

48. The oligonucleotide is WV-26758, WV-26759, WV-29875, WV-29876, WV-29877, WV-29878, WV-29879, WV-29880, WV-29881, WV-29882, WV-29883, WV-29884, WV-29885, WV-29886, WV-29887, WV-29888, WV-30672, WV-30673, WV-30674, WV-30970, WV-30 971, WV-30972, WV-30973, WV-30974, WV-30975, WV-30976, WV-30977, WV-30978, WV-30979, WV-30980, WV-30981, WV-309 82, WV-30983, WV-30984, WV-30985, WV-30986, WV-30987, WV-30988, WV-30989, WV-30990, WV-30991, WV-30992, WV-3099 3. WV-30994, WV-30995, WV-30996, WV-30997, WV-30998, WV-30999, WV-31000, WV-31001, WV-31002, WV-31003, WV-31004 , WV-31005, WV-31006, WV-31007, WV-31008, WV-31009, WV-31010, WV-31011, WV-31012, WV-31013, WV-31014, WV-31015, WV-31016, WV-31017, WV-31018, WV-31019, WV-31020, WV-31021, WV-31022, WV-31023, WV-31024, WV-31025, WV-31026, W V-31027, WV-31028, WV-31029, WV-31030, WV-31031, WV-31032, WV-31033, WV-31034, WV-31035, WV-31036, WV-31037, WV -3110338、WV-311039、WV-311040、WV-311041、WV-311042、WV-311043、WV-311044、W V-311045、WV-311046、WV-311048、WV-311049、WV-311050、WV-311051、WV-311052、 WV-3322808、WV-3322809、WV-33228110、WV-33228111、WV-33228112、WV-33228113、WV-33228114 、WV-322815、WV-322816、WV-323228117、WV-323228118、WV-32322819、WV-323228220、WV-3228221 、WV-328222、WV-3282223、WV-3328224、WV-3328225、WV-3328225、WV-3328225、WV-3328226、WV-3328227、WV-333300 8、WV-333009、WV-33330110、WV-33330111、WV-33330112、WV-33330113、WV-3333、WV-3330 15、WV-3333016、WV-33330117、WV-33330118、WV-333330119、WV-333330220、WV-3333 022、WV-33330203、WV-33330204、WV-33330205、WV-333330206、WV-333330207、WV-3333 0229、WV-3333030、WV-3333031、WV-3333032、WV-3333034、WV-3333035、WV-3 3036、WV-333037、WV-333038、WV-333039、WV-333040、WV-333041、WV-333042、WV- 333043、WV-3333044、WV-3333045、WV-3333046、WV-3333047、WV-3333048、WV-3333049、WV -3333050、WV-3333051、WV-3333052、WV-33333053、WV-3333054、WV-3333055、WV-3333056、WV -333057、WV-3333058、WV-3333059、WV-3333060、WV-3333061、WV-3333063、W V-333064、WV-333065、WV-3333066、WV-3333067、WV-3333068、WV-3333069、WV-3333070、W V-3333071、WV-3333072、WV-3333073、WV-3333074、WV-3333075、WV-3333076、WV-333077、 WV-33078、WV-33079、WV-33080、WV-33081、WV-33082、WV-33083、WV-33084、oligonucleotides that are WV-33085, WV-33086, WV-33087, WV-33088, WV-33089, WV-33090, WV-36875, WV-36876, WV-36877, WV-37299, WV-37300, WV-37301, WV-37302, WV-37303, WV-37304, or WV-37305.

49. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-29883.

50. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-32823.

51. The oligonucleotide according to claim 48, which is WV-29876.

52. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-32824.

53. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-32826.

54. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-29884.

55. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-29886.

56. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-29877.

57. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-32816.

58. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-32817.

59. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-29878.

60. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-29887.

61. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-32827.

62. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-32825.

63. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-29885.

64. The oligonucleotide according to claim 48, wherein the oligonucleotide is WV-29882.

65. An oligonucleotide according to any one of claims 48 to 64, which is in salt form.

66. An oligonucleotide according to any one of claims 48 to 64, which is a pharmaceutically acceptable salt form.

67. The oligonucleotide according to claim 66, which is in sodium salt form.

68. The oligonucleotide according to any one of claims 1 to 67, having a diastereomer purity of approximately 60% to 100%.

69. The oligonucleotide according to any one of claims 1 to 68, having a diastereomer purity of at least about 60%.

70. An oligonucleotide according to any one of claims 1 to 69, having the ability to reduce the level, expression, and / or activity of a MAPT transcript when administered to a system containing the MAPT transcript.

71. A pharmaceutical composition comprising delivering or comprising an oligonucleotide according to any one of claims 1 to 70 and a pharmaceutically acceptable carrier.

72. The composition according to claim 71, comprising delivering or comprising a pharmaceutically acceptable salt of the oligonucleotide.

73. The composition according to claim 71 or 72, comprising two or more forms of the oligonucleotide.

74. The composition according to any one of claims 71 to 73, which is an aqueous composition comprising one or more dissolved pharmaceutically acceptable salt forms of the oligonucleotide.

75. An oligonucleotide composition comprising a plurality of oligonucleotides, wherein one of the plurality of oligonucleotides is 1) Common base sequence, and 2) One or more (for example, about 1-50, 1-40, 1-30, 1-25, 1-20, 1-15, 1-10, 5-50, 5-40, 5-30, 5-25, 5-20, 5-15, 5-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 or more) chiral internucleotide bonds with independently identical binding phosphorus stereochemistry ("chiral-controlled internucleotide bonds") share; The oligonucleotide composition comprises several oligonucleotides, each independently targeting MAPT.

76. An oligonucleotide composition comprising a plurality of oligonucleotides, wherein one of the plurality of oligonucleotides is 1) Common base sequence, and 2) One or more (for example, about 1-50, 1-40, 1-30, 1-25, 1-20, 1-15, 1-10, 5-50, 5-40, 5-30, 5-25, 5-20, 5-15, 5-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 or more) chiral internucleotide bonds with independently identical binding phosphorus stereochemistry ("chiral-controlled internucleotide bonds") share; The oligonucleotide composition is characterized by having a higher purity of one of the oligonucleotides compared to a substantially racemic formulation of oligonucleotides sharing the common base sequence.

77. a) Common base sequence; b) Common skeletal connection patterns; c) Common skeletal chiral center pattern A composition comprising an oligonucleotide of a specific oligonucleotide type characterized by, The oligonucleotides among the multiple oligonucleotides include at least one internucleotide bond containing a common binding phosphorus in an Sp configuration; The composition is purified with respect to the specific oligonucleotide type compared to a substantially racemic formulation of oligonucleotides having the same common base sequence; The composition comprises several oligonucleotides, each independently targeting MAPT.

78. The composition according to claim 75 or 76, wherein the oligonucleotide has the ability to reduce the level, expression, and / or activity of the MAPT transcript when administered to a system containing the MAPT transcript.

79. The composition according to any one of claims 75 to 78, wherein the oligonucleotide hybridizes to a site in the MAPT transcript.

80. The composition according to any one of claims 75 to 79, wherein the oligonucleotide comprises a first wing, a core, and a second wing.

81. The composition according to any one of claims 75 to 80, wherein the sugar modification pattern of the first wing is different from the sugar modification pattern of the second wing.

82. The composition according to any one of claims 75 to 81, wherein the nucleoside unit of the core is not sugar-modified.

83. The composition according to any one of claims 75 to 82, wherein the oligonucleotide targets the MAPT intron 11.

84. The PLC targets the MAPT intron 11, and the PLC is ACGTTGCAGTGTTCCACUAU, ACTATCCTCTCTCCAGCUCCU, AGTGTTCCAACTATCCUCUCCUU, ATCCTCTCTCCAGCTCUGCA, CACGTTGCAGTGTTCCACUA, CACTATCCTCTCTCCAGCUC, CAGTGTTCCAACTATCCUCUCCU, CCACGTTGCAGTGTTCCACU, CCACTATCCTCTCTCCAGCUC, CGTTTGCAGTGTTCCACUAUUC, CTATCCTCTCTCCAGCUCUCUG, GCAGTGTTC A composition according to any one of claims 75 to 82, having a base sequence comprising at least 10 adjacent bases of the base sequence CACTATCCUCC, GTGTTCCCACTATCCTCCUUC, GTTCCCACTATCCTCCUUCAG, GUGUUCCCACTATCCTCTCTC, TATCTCTCTCCAGCTCCUGC, TCCAACTATCCTCTCTCCAGCCU, TGCAGTGTTCCCACTAUCCUC, TGTTCCACACTATCCTCCUUCA, TTCCACACTATCCTCTCTUCAGC, or TTGCCAGTGTTCCCACTAUCCCU (wherein each T can be independently substituted with U, and vice versa).

85. An oligonucleotide composition comprising a plurality of oligonucleotides, wherein one of the plurality of oligonucleotides is 1) Common base sequence, and 2) One or more (for example, about 1-50, 1-40, 1-30, 1-25, 1-20, 1-15, 1-10, 5-50, 5-40, 5-30, 5-25, 5-20, 5-15, 5-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 or more) chiral internucleotide bonds with independently identical binding phosphorus stereochemistry ("chiral-controlled internucleotide bonds") share; An oligonucleotide composition in which each of the plurality of oligonucleotides is independently an oligonucleotide according to any one of claims 1 to 67.

86. a) Common base sequence; b) Common skeletal connection patterns; c) Common skeletal chiral center pattern A composition comprising an oligonucleotide of a specific oligonucleotide type characterized by, The oligonucleotides among the multiple oligonucleotides include at least one internucleotide bond containing a common binding phosphorus in an Sp configuration; The composition is purified with respect to the specific oligonucleotide type compared to a substantially racemic formulation of oligonucleotides having the same common base sequence; A composition in which each of the plurality of oligonucleotides is independently an oligonucleotide according to any one of claims 1 to 67.

87. The composition according to any one of claims 75 to 86, wherein the plurality of oligonucleotides independently share the same binding phosphorus stereochemistry in 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 or more internucleotide bonds.

88. The composition according to claim 75 or 76, wherein the oligonucleotides among the plurality are oligonucleotides having the same chemical structure in one or more salt forms, which can be optionally selected.

89. The composition according to any one of claims 75 to 88, wherein each of the plurality of oligonucleotides is independently the same oligonucleotide or a pharmaceutically acceptable salt thereof.

90. The composition according to any one of claims 75 to 89, wherein each of the plurality of oligonucleotides is independently the same oligonucleotide or a pharmaceutically acceptable salt thereof.

91. The composition according to any one of claims 75 to 90, wherein the oligonucleotides among the plurality are one or more pharmaceutically acceptable salts of the same acid-form oligonucleotide.

92. The composition according to any one of claims 75 to 91, wherein the oligonucleotides among the plurality have the same structure.

93. The composition according to any one of claims 75 to 92, wherein the oligonucleotide among the plurality is a sodium salt.

94. The composition according to any one of claims 75 to 93, wherein the oligonucleotides among the plurality independently share the same bonding phosphorus stereochemistry at each phosphorothioate internucleotide bond.

95. The composition according to any one of claims 75 to 93, wherein the oligonucleotides among the plurality independently share the same bonded phosphorus stereochemistry at each chiral-bonded phosphorus.

96. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 48.

97. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 49.

98. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 50.

99. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 51.

100. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 52.

101. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 53.

102. Claims 75-86, each of the plurality of oligonucleotides is, independently and at will, an oligonucleotide according to claim 54 in a pharmaceutically acceptable salt form. A composition according to any one of the items.

103. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 55.

104. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 56.

105. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 57.

106. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 58.

107. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 59.

108. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 60.

109. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 61.

110. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 62.

111. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 63.

112. The composition according to any one of claims 75 to 86, wherein each of the plurality of oligonucleotides is independently and optionally in a pharmaceutically acceptable salt form, according to claim 64.

113. The composition according to any one of claims 75 to 112, wherein all non-random levels of oligonucleotides in the composition that share the common base sequence are oligonucleotides among the plurality.

114. The composition according to any one of claims 75 to 113, wherein all non-random levels of oligonucleotides in the composition that share the same chemical structure are oligonucleotides among the plurality.

115. The composition according to claim 113 or 114, wherein the non-random levels are approximately 40% to 100% or approximately 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.

116. A pharmaceutical composition comprising or delivering an oligonucleotide of the present disclosure or a composition according to any one of claims 75 to 115 and a pharmaceutically acceptable carrier.

117. A method for preparing an oligonucleotide or composition according to any one of claims 1 to 116, comprising the use of a chiral auxiliary group.

118. The method according to claim 117, comprising using a phosphoramidite containing a chiral auxiliary group.

119. The method according to claim 117 or 118, wherein the chiral auxiliary group is DPSE.

120. The method according to any one of claims 117 to 119, wherein the chiral auxiliary group is PSM.

121. The method according to any one of claims 117 to 120, wherein each chiral-controlled internucleotide bond, in which a bound phosphorus is linked to a nitrogen, is independently prepared using PSM.

122. The method according to any one of claims 117 to 121, wherein each chiral-controlled n001 is prepared independently using PSM.

123. The method according to any one of claims 117 to 122, wherein each chiral-controlled phosphorothioate internucleotide bond is prepared independently using DPSE.

124. The method according to claim 117 or 118, wherein each chiral-controlled internucleotide bond is prepared independently using PSM.

125. A method for reducing the level, function, and / or activity of a tau protein, comprising contacting the protein with an oligonucleotide or composition according to any one of claims 1 to 124.

126. A method for reducing intracellular aggregation of tau, comprising contacting the protein with an oligonucleotide or composition according to any one of claims 1 to 125.

127. A method for reducing the spread of tau, comprising contacting the protein with an oligonucleotide or composition according to any one of claims 1 to 126.

128. A method for reducing intracellular aggregation and spreading of tau, comprising contacting the protein with an oligonucleotide or composition according to any one of claims 1 to 127.

129. A method for reducing the level, function, and / or activity of tau protein in a system, comprising administering an effective amount of an oligonucleotide or composition according to any one of claims 1 to 128 to the system.

130. A method for reducing intracellular aggregation of tau in a system, comprising administering an effective amount of an oligonucleotide or composition according to any one of claims 1 to 129 to the system.

131. A method for reducing the spread of tau within a system, comprising administering an effective amount of an oligonucleotide or composition according to any one of claims 1 to 130 to the system.

132. A method for reducing intracellular aggregation and spreading of tau within a system, comprising administering an effective amount of an oligonucleotide or composition according to any one of claims 1 to 131 to the system.

133. The method according to any one of claims 129 to 132, wherein the system is human.

134. A method for treating or preventing a MAPT-related pathological condition, disorder, or disease or its symptoms in a subject who is suffering from or susceptible to such condition, comprising administering a therapeutically effective amount of the oligonucleotide or composition described in any one of claims 1 to 133 to the subject.

135. A method for treating or preventing a neurodegenerative disease in a subject who is suffering from or susceptible to such disease, comprising administering a therapeutically effective amount of the oligonucleotide or composition described in any one of claims 1 to 134 to the subject.

136. A method for treating or preventing tauopathy in a subject who is suffering from or susceptible to tauopathy, comprising administering a therapeutically effective amount of the oligonucleotide or composition described in any one of claims 1 to 135 to the subject.

137. The method according to any one of claims 134 to 136, wherein the aforementioned pathological condition, disorder, or disease is Alzheimer's disease (AD).

138. The method according to any one of claims 134 to 136, wherein the aforementioned pathological condition, disorder, or disease is frontotemporal dementia (FTD).

139. The method according to any one of claims 134 to 136, wherein the aforementioned pathological condition, disorder, or disease is behavioral FTD (bvFTD).

140. The method according to any one of claims 134 to 136, wherein the aforementioned pathological condition, disorder, or disease is non-fluent primary progressive aphasia (nfvPPA).

141. The method according to any one of claims 134 to 136, wherein the aforementioned pathological condition, disorder, or disease is corticobasal degeneration (CBD).

142. The method according to any one of claims 134 to 136, wherein the aforementioned pathological condition, disorder, or disease is progressive supranuclear palsy (PSP).

143. The method according to any one of claims 134 to 136, wherein the aforementioned pathological condition, disorder, or disease is epilepsy.

144. The method according to any one of claims 134 to 136, wherein the aforementioned pathological condition, disorder, or disease is Dravet syndrome.

145. The method according to any one of claims 134 to 136, wherein the aforementioned pathological condition, disorder, or disease is chronic traumatic encephalopathy (CTE).

146. Compounds, oligonucleotides, compositions, and methods described herein or in embodiments 1 to 145.

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