NMDA ligand conjugate compounds and their use

Abstract

The present invention provides compounds comprising an N-methyl-D-aspartate (NMDA) receptor ligand, methods for delivering the compound, and methods for targeting diseases, disorders, and symptoms (e.g., diseases, disorders, and symptoms of the central nervous system) using the compound. Specific embodiments provided in the present invention relate to compounds and methods for delivering drugs to cells expressing the NMDA receptor. Specific embodiments provided in the present invention relate to compounds and methods for regulating the expression of nucleic acid targets in cells expressing the NMDA receptor.

Description

[Technical Field]

[0001] Related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 502,035, filed on 12 May 2023 pursuant to Section 119(e) of the U.S. Patent Act, the contents of which are incorporated herein by reference. [Background technology]

[0002] When using compounds for therapeutic, preventive, or diagnostic purposes, it is often desirable to deliver them to specific locations (e.g., desired cells) to enhance therapeutic or preventive effects, or to be beneficial for diagnostic purposes. This is a common scenario when attempting to deliver therapeutic compounds in vivo. Furthermore, efficient delivery of compounds to specific locations can limit or potentially eliminate unintended consequences (such as off-target effects) that may result from the administration of the compound. One strategy to facilitate the in vivo delivery of compounds, such as therapeutic, preventive, or diagnostic compounds, to desired locations is to link or bind them to a targeting ligand.

[0003] One type of compound that can be derived using targeting ligands is oligomeric compounds, such as proteins, peptides, antibodies, and oligonucleotides. Oligomer compounds containing a nucleotide sequence (e.g., oligonucleotide) at least partially complementary to a target nucleic acid have been shown to alter the function and activity of the target both in vitro and in vivo. Oligonucleotides have been shown to modulate the expression or activity of a target nucleic acid (such as mRNA or premRNA) when delivered to cells containing that target nucleic acid. In certain cases, the oligonucleotide can reduce the expression of its gene by inhibiting the translation of its nucleic acid target and / or inducing the degradation of that target nucleic acid.

[0004] When the target nucleic acid is mRNA, one mechanism by which oligonucleotides can regulate the expression of its mRNA target is through RNA interference. RNA interference is a biological process in which RNA or RNA-like molecules (such as chemically modified RNA molecules) can at least partially silence gene expression through the RNA-induced silencing complex (RISC) pathway. In addition, oligonucleotides can regulate the expression of target nucleic acids, such as target mRNA, through RNase recruitment mechanisms, microRNA mechanisms, occupation-based mechanisms, and editing mechanisms. Oligonucleotides may be single-stranded or double-stranded. Oligonucleotides may contain DNA, RNA, and RNA-like molecules, and may also contain one or more modified sugars, modified nucleic acid bases, and modified nucleosides containing modified nucleoside-to-nucleoside bonds.

[0005] Another type of compound that can be directed using targeting ligands is small molecule compounds. These small molecule compounds (e.g., organic compounds with a molecular weight of approximately 1000 daltons or less) have typically been shown to modulate or improve disease and / or disease symptoms by altering the function and / or activity of the target when localized to it, or to be useful as a diagnostic marker. More efficient delivery of compounds to specific locations can limit or potentially eliminate unintended consequences (such as off-target effects) that may result from the administration of the compound, and can also improve the localization of diagnostic compounds. [Overview of the project] [Means for solving the problem]

[0006] The embodiments provided in this invention relate to compounds (e.g., any of the compounds revealed in this invention) and methods for targeting cells expressing N-methyl-D-aspartate (NMDA) receptors. Specific embodiments provided in this invention relate to compounds and methods for delivering a drug to cells expressing NMDA receptors. In specific embodiments, the cells are located in the brain. In specific embodiments, the cells are located in the prefrontal cortex. In specific embodiments, the cells are located in the striatum. In specific embodiments, the cells are located in the cerebellum. In specific embodiments, the cells are located in the brainstem. In specific embodiments, the cells are located in the hippocampus. In specific embodiments, the cells are located in the spinal cord. In specific embodiments, the drug is a therapeutic compound. In specific embodiments, the delivery of the drug is for the treatment of diseases, disorders, and symptoms in a subject. In specific embodiments, the drug is a diagnostic compound. In specific embodiments, the compound comprises an NMDA receptor ligand and one or more linker moieties for binding to a therapeutic, prophylactic, or diagnostic agent. In specific embodiments, the compound comprises an NMDA receptor ligand, one or more linker moieties, and a therapeutic agent. In certain embodiments, the therapeutic agent is selected from small molecules or oligomeric compounds. In certain embodiments, the oligomeric compound is a protein, peptide, antibody, oligonucleotide, or a combination thereof. In certain embodiments, the NMDA receptor ligand is an NMDA receptor agonist. In certain embodiments, the NMDA receptor ligand is an NMDA receptor antagonist. In certain embodiments, the NMDA receptor ligand is a small molecule, aptamer, peptide, or antibody. In certain embodiments, the NMDA receptor ligand is any of those revealed in the present invention, or a derivative thereof, or a prodrug.

[0007] In certain embodiments, contact between cells expressing NMDA receptors, such as brain cells, and the compounds provided in this invention results in the delivery of the drug to those cells. In certain embodiments, contact between cells expressing NMDA receptors, such as brain cells, and the compounds provided in this invention results in the targeted treatment of a disease, disorder, or symptom. In certain embodiments, compounds containing an NMDA receptor ligand selectively or preferentially target cells expressing NMDA receptors compared to cells that do not express NMDA receptors. In certain embodiments, compounds containing an NMDA receptor ligand selectively or preferentially target cells expressing NMDA receptors compared to compounds that do not contain an NMDA receptor ligand.

[0008] The specific embodiments provided in this invention relate to compounds and methods for regulating the expression of nucleic acid targets in cells expressing NMDA receptors. In specific embodiments, the cells are located in the brain. In specific embodiments, the cells are located in the prefrontal cortex. In specific embodiments, the cells are located in the striatum. In specific embodiments, the cells are located in the cerebellum. In specific embodiments, the cells are located in the brainstem. In specific embodiments, the cells are located in the hippocampus. In specific embodiments, the cells are located in the spinal cord. In specific embodiments, contact between cells expressing NMDA receptors, such as brain cells, and the compounds provided in this invention results in regulation of the expression or activity of nucleic acid targets in those cells. In specific embodiments, the compound comprises an NMDA receptor ligand, one or more linker moieties, and an oligonucleotide.

[0009] It should be understood that the embodiments provided in this invention may take the form of a single entity or a combination of one or more embodiments or other preferred variables provided in this invention, as if each combination were explicitly enumerated herein.

[0010] In one embodiment, the present disclosure provides compounds of the following formula (I) and salts thereof, [ka] During the ceremony, [ka] It is an N-methyl-D-aspartate (NMDA) receptor ligand, Each of L1, L2, L3, and L4 is independently either a linker (e.g., an optionally substituted alkyl linker, an optionally substituted polyethylene glycol (PEG) linker, an optionally substituted heteroalkyl linker, or an optionally substituted heteroaryl linker), a bond (e.g., an intercarbon bond, a phosphodiester bond, or a phosphorothioate bond), or absent. Y is a bond or -C(=O)-, R 1 This is one or more oligonucleotides, protecting groups, small molecules, proteins, antibodies, and / or peptides.

[0011] In some embodiments, the NMDA receptor ligand is an NMDA receptor agonist. In some embodiments, the NMDA receptor ligand is an NMDA receptor antagonist. In some embodiments, the NMDA receptor ligand is [ka] The antibodies are selected from the group consisting of anti-NMDA receptor antibodies and their derivatives.

[0012] In some embodiments, the Disclosure provides compounds comprising the structures of the following formula (II) and its salts, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0013] In some embodiments, the compound comprises the structure of the following formula (II-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I).

Chem.

[0014] In some embodiments, the present disclosure provides compounds comprising the structure of formula (III) below and salts thereof, wherein Y, L1, L2, L3, L4, and R 1 are as defined in formula (I), and R 2 is hydrogen, halogen, -OH, or -OMe.

Chem.

[0015] In some embodiments, the present disclosure provides compounds comprising the structure of formula (III-a) below and salts thereof, wherein Y, L1, L2, L3, L4, and R 1 are as defined in formula (I), and R 2 is hydrogen, halogen, -OH, or -OMe.

Chem.

[0016] In some embodiments, the compound comprises the structure of formula (III-b) below or a salt thereof, wherein Y, L1, L2, L3, L4, and R 1 are as defined in formula (I).

Chem.

[0017] In some embodiments, the compound comprises the structure of formula (III-c) below or a salt thereof, wherein Y, L1, L2, L3, L4, and R 1 are as defined in formula (I).

Chem.

[0018] In some embodiments, the compound comprises the structure of the following formula (III-d) or a salt thereof, where Y, L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0019] In some embodiments, the compound comprises the structure of the following formula (III-e) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0020] In some embodiments, the compound comprises the structure of the following formula (III-f) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0021] In some embodiments, the compound comprises the structure of the following formula (III-g) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0022] In some embodiments, the compound comprises the structure of the following formula (III-h) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0023] In some embodiments, the compound comprises the structure of the following formula (III-i) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0024] In some embodiments, the compound comprises the structure of the following formula (III-j) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0025] In some embodiments, the compound comprises the structure of the following formula (III-k) or a salt thereof, where Y, L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0026] In some embodiments, the compound comprises the structure of the following formula (III-l) or a salt thereof, where Y, L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0027] In some embodiments, the compound comprises the structure of the following formula (III-m) or a salt thereof, where Y, L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0028] In some embodiments, the compound comprises the structure of the following formula (III-n) or a salt thereof, where Y, L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0029] In some embodiments, the compound comprises the structure of the following formula (III-o) or a salt thereof, where Y, L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0030] In some embodiments, the compound comprises the structure of the following formula (III-p) or a salt thereof, where Y, L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0031] In some embodiments, the Disclosure provides compounds comprising the structures of the following formula (IV) and its salts, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0032] In some embodiments, the compound comprises the structure of the following formula (IV-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0033] In some embodiments, the compound comprises the structure of the following formula (XIX) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0034] In some embodiments, the compound comprises the structure of the following formula (XIX-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0035] In some embodiments, the present disclosure provides compounds comprising the structures of formula (V) and its salts, wherein the formula L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0036] In some embodiments, the compound comprises the structure of the following formula (Va) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0037] In some embodiments, the present disclosure provides compounds comprising the structures of formula (VI) and its salts, wherein L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0038] In some embodiments, the compound comprises the structure of the following formula (VI-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0039] In some embodiments, the compound comprises the structure of the following formula (VI-b) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0040] In some embodiments, the present disclosure provides compounds comprising the structures of formula (VII) and its salts, wherein L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0041] In some embodiments, the compound comprises the structure of the following formula (VII-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0042] In some embodiments, the present disclosure provides compounds comprising the structures of formula (VIII) and its salts, wherein L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0043] In some embodiments, the compound comprises the structure of the following formula (VIII-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0044] In some embodiments, the compound comprises the structure of the following formula (IX) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0045] In some embodiments, the compound comprises the structure of the following formula (IX-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0046] In some embodiments, the compound comprises the structure of the following formula (X) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0047] In some embodiments, the compound comprises the structure of the following formula (Xa) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0048] In some embodiments, the compound comprises the structure of the following formula (Xb) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0049] In some embodiments, the compound comprises the structure of the following formula (XI) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0050] In some embodiments, the compound comprises the structure of the following formula (XI-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0051] In some embodiments, the compound comprises the structure of the following formula (XI-b) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0052] In some embodiments, the compound comprises the structure of the following formula (XI-c) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0053] In some embodiments, the compound comprises the structure of the following formula (XII) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0054] In some embodiments, the compound comprises the structure of the following formula (XII-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0055] In some embodiments, the compound comprises the structure of the following formula (XII-b) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0056] In some embodiments, the compound comprises the structure of the following formula (XIII) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0057] In some embodiments, the compound comprises the structure of the following formula (XIII-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0058] In some embodiments, the compound comprises the structure of the following formula (XIV) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0059] In some embodiments, the compound comprises the structure of the following formula (XIV-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0060] In some embodiments, the compound comprises the structure of the following formula (XV) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0061] In some embodiments, the compound comprises the structure of the following formula (XV-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0062] In some embodiments, the compound comprises the structure of the following formula (XVI) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0063] In some embodiments, the compound comprises the structure of the following formula (XVI-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0064] In some embodiments, the compound comprises the structure of the following formula (XVII) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0065] In some embodiments, the compound comprises the structure of the following formula (XVII-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0066] In some embodiments, the compound comprises the structure of the following formula (XVIII) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0067] In some embodiments, the compound comprises the structure of the following formula (XVIII-a) or a salt thereof, where L1, L2, L3, L4 and R 1 This is as defined in equation (I). [ka]

[0068] In some embodiments, any one of L1, L2, L3, and L4 may independently be a linker (e.g., an optionally substituted alkyl linker, an optionally substituted polyethylene glycol (PEG) linker, an optionally substituted heteroalkyl linker, or an optionally substituted heteroaryl linker). In some embodiments, any one of L1, L2, L3, and L4 may independently be a bond (e.g., a carbon-carbon bond, a phosphodiester bond, or a phosphorothioate bond). In some embodiments, any one of L1, L2, L3, and L4 may independently be absent.

[0069] In some embodiments, L1 is a bond. In some embodiments, L1 is an optionally substituted alkyl linker. In some embodiments, L1 is an optionally substituted C1-C6 alkyl linker. In some embodiments, L1 is a C1-C6 alkyl linker substituted with =O. In certain embodiments, L1 is

Chemical formula

[0070] In some embodiments, L2 is an optionally substituted alkyl linker. In some embodiments, L2 is an optionally substituted C1-C 15 alkyl linker. In some embodiments, L2 is an optionally substituted C5-C 12 alkyl linker. In certain embodiments, L2 is

Chemical formula

Chemical formula

[0071] In some embodiments, L2 is an optionally substituted PEG linker. In some embodiments, L2 is an optionally substituted PEG linker having a PEG unit length of 1, 2, 3, 4, 5, 6, 7 or 8, and the PEG unit is [Chemical formula] including the structure of. In certain embodiments, L2 is an optionally substituted PEG linker having a PEG unit length of 3. In certain embodiments, L2 is an optionally substituted PEG linker having a PEG unit length of 4. In certain embodiments, L2 is [Chemical formula] including the structure of.

[0072] In some embodiments, L2 is an optionally substituted heteroalkyl linker. In certain embodiments, L2 is [Chemical formula] including the structure of.

[0073] In some embodiments, L3 is an optionally substituted heteroaryl linker. In some embodiments, L3 is an optionally substituted partially unsaturated heterocycloalkyl linker or heteroaryl linker. In certain embodiments, L3 is [Chemical formula] including the structure of.

[0074] In some embodiments, L4 is an optionally substituted heteroalkyl linker. In some embodiments, the heteroalkyl linker is substituted with one or more =O substituents. In certain embodiments, L4 is [Chemical formula] including the structure, wherein X is O or S. In certain embodiments, L4 is [Chemical formula] including the structure, wherein X is O or S.

[0075] In some embodiments, L1, L2, L3 and L4 are integrated into [Chemical formula] including the structure, wherein X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated into [Chemical formula] including the structure, wherein X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated into [Chemical formula] including the structure, wherein X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated into [Chemical formula] including the structure, wherein X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated into [Chemical formula] including the structure, wherein X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated into [Chemical formula] <​​​​​​​​ The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes the form where X is either O or S.

[0076] In certain embodiments, this disclosure is, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present disclosure provides compounds having the structure of the salt thereof, where X is O or S. In certain embodiments, the present disclosure provides, [ka] The present invention provides compounds having the structure of a salt thereof, where X is O or S.

[0077] In some embodiments, the present disclosure provides compounds comprising the structure of the following formula (XX): [ka] During the ceremony, each [ka] It is an N-methyl-D-aspartate (NMDA) receptor ligand, L1, L2, L3, L4, L 1a , L 2a , L 3a and L 4a Each of these can independently be a linker (e.g., an optionally substituted alkyl linker, an optionally substituted polyethylene glycol (PEG) linker, an optionally substituted heteroalkyl linker, or an optionally substituted heteroaryl linker), a bond (e.g., an intercarbon bond, a phosphodiester bond, or a phosphorothioate bond), or may not be present. Y and Y a Each of them is independently a bond or -C(=O)-, [ka] It is an oligonucleotide.

[0078] In some embodiments, the present disclosure provides compounds comprising the structure of formula (XXI) below, [ka] During the ceremony, [ka] Each of these is independently an N-methyl-D-aspartate (NMDA) receptor ligand, L1, L2, L3, L4, L 1a , L 2a , L 3a and L 4a Each of these independently is either a linker (e.g., an optionally substituted alkyl linker, an optionally substituted polyethylene glycol (PEG) linker, an optionally substituted heteroalkyl linker, or an optionally substituted heteroaryl linker), a bond (e.g., an intercarbon bond, a phosphodiester bond, or a phosphorothioate bond), or is absent. Y and Y a Each of them is independently a bond or -C(=O)-, [ka] It is an oligonucleotide.

[0079] In certain embodiments, this disclosure is, [ka] [ka] [ka] The present invention provides a compound containing the structure, where each instance of X is independently either O or S. [ka] It is an oligonucleotide.

[0080] In some embodiments, X is O. In some embodiments, X is S.

[0081] In some embodiments, R 1 It contains an oligonucleotide. In some embodiments, the oligonucleotide is bonded at its 5' end. In some embodiments, the oligonucleotide is bonded at its 3' end. In some embodiments, the oligonucleotide is bonded at an internal position on the oligonucleotide. In some embodiments, the internal position is at an internucleoside bond. In some embodiments, R 1 This comprises an oligonucleotide conjugated with one or more additional NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated with two, three, four, or five or more additional NMDA receptor ligands. In certain embodiments, the additional NMDA receptor ligands are conjugated to the oligonucleotide at one or more of the 5' end, the 3' end, one or more internal positions on the oligonucleotide, or a combination thereof. In certain embodiments, the oligonucleotide is a modified oligonucleotide.

[0082] In another aspect, the present disclosure provides a composition comprising one of the compounds provided in the present invention and a pharmaceutically acceptable excipient.

[0083] In another embodiment, the Disclosure provides a method for delivering a therapeutic oligonucleotide to a target brain, comprising administering one of the compounds or compositions provided in the Invention to the target. In some embodiments, the therapeutic oligonucleotide is delivered to one or more brain regions selected from the group consisting of the striatum, cerebellum, brainstem, hippocampus, prefrontal cortex, and spinal cord. In another embodiment, the Disclosure provides a method for treating or improving a disease, disorder, or symptom in a target, comprising administering one of the compounds or compositions provided in the Invention to the target. In some embodiments, the disease, disorder, or symptom is a disease, disorder, or symptom of the central nervous system (CNS). In certain embodiments, the disease, disorder, or symptom is Alzheimer's disease or a symptom of Alzheimer's disease. In some embodiments, the compound is administered intrathecally to the target.

[0084] In another aspect, the present disclosure provides a method for producing any of the compounds provided in the present invention, comprising one or more compounds and chemical conversions described herein (including Examples 1 to 77). [Modes for carrying out the invention]

[0085] definition It should be understood that both the general description above and the detailed description below are illustrative and descriptive, and do not limit the embodiments claimed. Where the singular form is used herein, it includes the plural unless otherwise specifically indicated. Where "or" is used herein, it means "and / or" unless otherwise indicated. Furthermore, where the term "including," and other forms such as "include" and "included," are used, they are not limiting. Section headings used herein are for structural purposes only and should not be construed as limiting the subject matter described.

[0086] Unless otherwise specified, the following terms have the following meanings:

[0087] As used herein, the term “treating” a disorder includes improving, mitigating, and / or managing the disorder and / or the conditions that may cause the disorder. The terms “treating” and “treatment” refer to methods of alleviating or reducing a disease and / or its associated symptoms. According to this disclosure, “treating” includes, for example, blocking the adverse effects of a disorder, inhibiting the adverse effects of a disorder, attenuating the adverse effects of a disorder, protecting from the adverse effects of a disorder, modulating the adverse effects of a disorder, reversing the adverse effects of a disorder, and reducing the occurrence of the adverse effects of a disorder. As used herein, “inhibiting” includes preventing, reducing, and stopping the progression of a disorder.

[0088] The terms “isolated,” “purified,” or “biologically pure” refer to a substance that substantially or essentially contains no components that would normally accompany it in its natural state. Purity and homogeneity are typically determined using analytical chemical methods, such as polyacrylamide gel electrophoresis or high-performance liquid chromatography (HPLC). In particular, in certain embodiments, the compound is at least 85% pure, more preferably at least 90% pure, more preferably at least 95% pure, and most preferably at least 99% pure.

[0089] The terms "administration" or "administration" include the route through which the compound(s) are introduced to the target in order to exert their intended function. Examples of usable routes of administration include injection (subcutaneous, intravenous, parenteral, intraperitoneal, intrathecal), topical, oral, inhalation, rectal, and percutaneous.

[0090] The term "effective dose" includes the amount that is effective in terms of dosage and duration required to achieve the desired result. The effective dose of a compound may vary depending on the factors that induce the desired response in the subject, such as the subject's condition, age, and weight, as well as the compound's capacity. The dosage regimen may be adjusted to produce the optimal therapeutic response. The effective dose is also the amount in which the therapeutically beneficial effects of the compound outweigh any tolerable or harmful effects (e.g., side effects).

[0091] The terms “systemic administration,” “administer systemically,” “peripheral administration,” and “administer peripherally” mean, as used herein, compounds (plural), oligonucleotides (plural), drugs, or other substances administered so that they enter the patient’s circulatory system and are subsequently subjected to metabolic processes and other similar processes.

[0092] The term "therapeutic dose" refers to a dosage of a compound sufficient to prevent or, to some extent, alleviate the occurrence of one or more symptoms of the condition or disease being treated.

[0093] The therapeutically effective dose (i.e., effective dosage) of the compound may be in the range of about 0.005 μg / kg to about 200 mg / kg of body weight, preferably about 0.01 mg / kg to about 200 mg / kg, more preferably about 0.015 mg / kg to about 30 mg / kg. In other embodiments, the therapeutically effective dose may be in the range of about 1.0 pM to about 10 μM. It will be apparent to those skilled in the art that certain factors may influence the dosage required to effectively treat a subject, including, but not limited to, the severity of the disease or disorder, previous treatments, the subject's overall health and / or age, and other pre-existing diseases. Furthermore, treating a subject with a therapeutically effective dose of the compound may include a single treatment, or preferably a series of treatments. In one example, the subject is treated with the compound daily, weekly, monthly, four times a year, or annually, in the range of about 0.005 μg / kg to about 200 mg / kg of body weight. In another example, the subjects may be treated daily, weekly, monthly, four times a year, or annually for several years in a chronic condition or disease state. It will also be clear that the effective dose of the compound used in treatment may be increased or decreased during a particular treatment.

[0094] The term "chiral" refers to a molecule that cannot be superimposed on its mirror image partner, while the term "achiral" refers to a molecule that can be superimposed on its mirror image partner.

[0095] Certain compounds in this disclosure have an asymmetric carbon atom (optical or chiral center) or a double bond, and the scope of this disclosure includes enantiomers, racemic compounds, diastereomers, tautomers, geometric isomers, stereoisomer forms that may be defined in terms of absolute stereochemistry, such as (R) or (S) or (D) or (L) forms for amino acids, and individual isomers. The compounds in this disclosure do not include compounds that are known in the art to be unstable and unsynthesizable and / or unisolated. This disclosure is intended to include compounds in racemic and optically pure forms. Optically active (R) and (S) or (D) and (L) isomers may be prepared using chiral synthons or chiral reagents, or divided using conventional techniques. When a compound described herein contains an olefin bond or other geometrically asymmetric center, and unless otherwise specified, the compound is intended to include both E and Z geometric isomers.

[0096] As used herein, the term "tautomer" refers to one of two or more structural isomers that exist in equilibrium and are readily convertible from one isomeric form to the other.

[0097] It will be apparent to those skilled in the art that certain compounds of this disclosure may exist in tautomeristic forms, and that any such tautomeristic forms of such compounds fall within the scope of this disclosure.

[0098] Unless otherwise indicated, the structures described herein are intended to include all stereochemical forms (i.e., R and S configurations for each chiral center). Accordingly, single stereochemical isomers of the compounds of the present invention, as well as enantiomer mixtures and diastereomer mixtures, are within the scope of this disclosure.

[0099] As used herein, “chiral-enriched population” means a group of molecules having the same molecular formula, where, if a particular chiral center is stereorandom, the number or percentage of molecules in the population containing a particular stereochemical configuration at that particular chiral center is greater than the number or percentage of molecules in the population that are likely to contain the same particular stereochemical configuration at the same particular chiral center. A group of molecules in a chiral-enriched population, each having multiple chiral centers, may contain one or more stereorandom chiral centers. In certain embodiments, the molecule is a modified oligonucleotide. In certain embodiments, the molecule is a compound containing a modified oligonucleotide.

[0100] Unless otherwise indicated, the structures described herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, hydrogen may be replaced with deuterium or tritium, or carbon may be replaced with 13 C concentrated carbon or 14 Compounds having the structure of the present invention, except that they are replaced with 13C-enriched carbon, are within the scope of this disclosure.

[0101] As used herein, “stereorandom chiral center” means a chiral center having a random stereochemical configuration with respect to a group of molecules with the same molecular formula. For example, in a group of molecules containing stereorandom chiral centers, the number of molecules having stereorandom chiral centers in (S) configuration may be the same as, but not necessarily the same as, the number of molecules having stereorandom chiral centers in (R) configuration. The stereochemical configuration of a chiral center is considered random when it is the result of a synthetic method not designed to control its stereochemical configuration. In certain embodiments, a stereorandom chiral center is a stereorandom phosphorothioate nucleoside bond.

[0102] The term "diastereomer" refers to stereoisomers that have two or more asymmetric centers and whose molecules are not mirror images of each other.

[0103] The term "enantiomer" refers to two stereoisomers of a compound that cannot be superimposed as mirror images of each other. An equimolar mixture of two enantiomers is called a "racemic mixture" or "racemic compound."

[0104] The terms "isomer" or "stereoisomer" refer to compounds that have the same chemical structure but differ in the arrangement of atoms or groups in space.

[0105] The term "prodrug" is intended to refer to a compound that can be converted under physiological conditions or by solvolysis to a biologically active form of the compound described herein (e.g., a biologically active form of nucleic acid) or an analogue thereof. Therefore, the term "prodrug" refers to a pharmaceutically acceptable precursor of a biologically active compound (e.g., nucleic acid) or an analogue thereof. Prodrugs may be inactive when administered to a subject but are converted in vivo to an active compound, for example, by hydrolysis. Prodrug compounds often offer advantages in mammalian organisms such as solubility, histocompatibility, or delayed release (see, e.g., Bundgaard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)). The discussion of prodrugs is presented in Higuchi, T., et al., “Pro-drugs as Novel Delivery Systems,” ACSSymposium Series, Vol.14 and Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference. The term “prodrug” is also intended to include any covalent carrier that, when such a prodrug is administered to a mammalian subject, releases the active compound in vivo. Prodrugs of active compounds, as described herein, can be prepared by modifying the functional groups present in the active compound in such a way that they are cleaved either by conventional methods or in vivo to become the parental active compound. Prodrugs include compounds in which a hydroxyl group, an amino group, or a mercapto group is bonded to one of these groups, and when the prodrug of the active compound is administered to a mammalian subject, it is cleaved to form a free hydroxyl group, a free amino group, or a free mercapto group, respectively.Suitable examples of prodrugs include, but are not limited to, glutathione derivatives, acyloxy derivatives, thioacyloxy derivatives, 2-carbokoxyethyl derivatives, disulfide derivatives, thiamine derivatives, and enol ester derivatives of phosphorus-modified nucleic acids. The terms “prooligonucleotide,” “pronucleotide,” or “nucleic acid prodrug” refer to an oligonucleotide that has been modified to be a prodrug of that oligonucleotide. Prodrugs of phosphonates and phosphates can be found, for example, in Wiener et al., “prodrugs or phosphonates and phosphates: crossing the membrane,” Top.Curr. Chem. 2015, 360:115-160, which is incorporated herein by reference in its entirety. Prodrugs that are converted to an active form in vivo through other mechanisms are also included. In some embodiments, the compounds of this disclosure are prodrugs of any of the formulas herein.

[0106] The term "subject" refers to animals, such as mammals, including but not limited to primates (e.g., humans), cattle, sheep, goats, horses, dogs, cats, rabbits, rats, and mice. In certain embodiments, the subject is a human.

[0107] The terms "a," "an," and "the" as used in this application (including the claims) refer to "one or more." Therefore, when "sample" is mentioned, for example, it includes multiple samples unless the context clearly indicates otherwise (e.g., multiple samples).

[0108] Throughout this specification and the claims, the terms “comprise,” “comprises,” and “comprising” are used non-exclusively unless the context requires otherwise.

[0109] As used herein, the term “about” is intended to include variations of ±20% in some embodiments, ±10% in some embodiments, ±5% in some embodiments, ±1% in some embodiments, ±0.5% in some embodiments, and ±0.1% in some embodiments from the specified amount. However, such variations shall be appropriate for carrying out the disclosed method or utilizing the disclosed composition.

[0110] As used herein, the term "alkyl" means, unless otherwise specified, a straight (i.e., unbranched) or branched carbon chain (or carbon) or combination thereof, either alone or as part of another substituent, and may be a fully saturated alkyl, a monoalkyl (e.g., an alkene or alkenyl), or a polyunsaturated alkyl (e.g., an alkyne or alkynyl), and may include monoradicals, diradicals, and polyradicals with a defined number of carbon atoms. For example, C1-C 24 This refers to 1 to 24 carbon atoms. Examples of carbon atoms within this range include C1-C 20 Alkyl (having 1 to 20 carbon atoms), C1-C 12 Examples include alkyl (having 1 to 12 carbon atoms) and C1-C4 alkyl (having 1 to 4 carbon atoms).

[0111] The term "alkenyl" refers to an unsaturated hydrocarbon chain, which may be straight or branched, containing 2 to 12 carbon atoms and at least one carbon-carbon double bond. The alkenyl group may be optionally substituted with one or more substituents.

[0112] The term "alkynyl" refers to an unsaturated hydrocarbon chain, which may be straight or branched, containing 2 to 12 carbon atoms and at least one carbon-carbon triple bond. The alkynyl group may be optionally substituted with one or more substituents.

[0113] The term "lower alkyl" refers to C1-C6 alkyl chains. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, tert-butyl, and n-pentyl. Alkyl groups may be optionally substituted with one or more substituents.

[0114] The term "heteroalkyl," unless otherwise indicated, means, alone or in combination with another term, a stable linear or branched chain, or a combination thereof, comprising at least one carbon atom and at least one heteroatom (e.g., O, N, P, Si, and / or S), wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) (e.g., O, N, P, Si, and / or S) may be located at any position within the heteroalkyl group or at a position where the alkyl group is bonded to the remainder of the molecule. A heteroalkyl group is an acyclized chain. Examples include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, -O-CH3, -O-CH2-CH3, and -CN. For example, up to two or three heteroatoms may follow, such as -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. The heteroalkyl portion may contain one heteroatom (e.g., O, N, S, Si, B, or P). The heteroalkyl portion may contain two arbitrarily distinct heteroatoms (e.g., O, N, S, Si, B, and / or P). The heteroalkyl portion may contain three arbitrarily distinct heteroatoms (e.g., O, N, S, Si, B, and / or P). The heteroalkyl portion may contain four arbitrarily distinct heteroatoms (e.g., O, N, S, Si, B, and / or P). The heteroalkyl portion may contain five arbitrarily distinct heteroatoms (e.g., O, N, S, Si, B, and / or P). The heteroalkyl portion may contain up to eight or more arbitrarily distinct heteroatoms (e.g., O, N, S, Si, B, and / or P).

[0115] Similarly, the term “heteroalkylene,” unless otherwise indicated, refers to a divalent radical derived from a heteroalkyl group, either alone or as part of another substituent, with examples including, but not limited to, -CH2-CH2-S-CH2-CH2- and -CH2-S-CH2-CH2-NH-CH2-. In heteroalkylene groups, the heteroatom can occupy either or both of its chain ends (e.g., alkylene oxy, alkylenedioxy, alkylene amino, alkylenediamino, etc.). Furthermore, in alkylene and heteroalkylene linking groups, the orientation of the linking group is not indicated by the direction in which the formula of the linking group is written. For example, the formula -C(O)2R'- represents both -C(O)2R'- and -R'C(O)2-. As stated above, as used herein, a heteroalkyl group includes groups bonded to the remainder of its molecule through heteroatoms, such as -C(O)R', -C(O)NR', ​​-NR'R'', -OR', -SR', and / or -SO2R'. Where “heteroalkyl” is indicated and followed by a list of specific heteroalkyl groups, such as -NR'R'', it should be understood that the term heteroalkyl and -NR'R'' are not redundant or mutually exclusive. Rather, the specific heteroalkyl group is shown for further clarification. Therefore, the term “heteroalkyl” should not be interpreted herein as excluding specific heteroalkyl groups, such as “-NR'R''.

[0116] The term "haloalkyl" refers to an alkyl group that is substituted with one or more halo substituents. Examples of haloalkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, bromomethyl, chloromethyl, and 2,2,2-trifluoroethyl.

[0117] The term "arylalkenyl" may include a straight or branched chain, and is an unsaturated hydrocarbon chain containing 2 to 12 carbon atoms and at least one carbon-carbon double bond, wherein the alkenyl unit is sp 2This refers to an unsaturated hydrocarbon chain in which one or more hybridized carbons are bonded to the aryl moiety. The alkenyl group may be optionally substituted with one or more substituents.

[0118] The term "arylalkynyl" refers to an unsaturated hydrocarbon chain, which may include a straight chain or a branched chain, containing 2 to 12 carbon atoms and at least one carbon-carbon triple bond, wherein one or more sp hybrid carbons of the alkynyl unit are bonded to the aryl portion. The alkynyl group may be optionally substituted with one or more substituents.

[0119] sp of alkenyl group and alkynyl group 2 The hybridized carbon or sp hybridized carbon may each be a bonding site of its alkenyl group or alkynyl group.

[0120] The term "alkoxy" refers to an -O-alkyl substituent.

[0121] As used herein, the terms "halogen," "hal," or "halo" mean -F, -Cl, -Br, or -I.

[0122] The term "alkylthio" refers to an -S-alkyl substituent.

[0123] The term "alkoxyalkyl" refers to an alkyl-O-alkyl substituent.

[0124] The term "haloalkoxy" refers to an -O-alkyl group substituted with one or more halo substituents. Examples of haloalkoxy groups include trifluoromethoxy and 2,2,2-trifluoroethoxy.

[0125] The term "haloalkoxyalkyl" refers to an alkyl-O-alkyl' group in which the alkyl' is substituted by one or more halo substituents.

[0126] The term "haloalkylaminocarbonyl" refers to a -C(O)-amino-alkyl group in which the alkyl group is substituted by one or more halo substituents.

[0127] The term "haloalkylthio" refers to an -S-alkyl group substituted with one or more halo substituents. Examples of haloalkylthio groups include trifluoromethylthio and 2,2,2-trifluoroethylthio.

[0128] The term "haloalkylcarbonyl" refers to a -C(O)-alkyl group that is substituted with one or more halo substituents. An example of a haloalkylcarbonyl group is trifluoroacetyl.

[0129] The term "cycloalkyl" refers to a 3- to 8-membered monocyclic hydrocarbon ring system or a 7- to 14-membered bicyclic hydrocarbon ring system having at least one saturated ring or at least one non-aromatic ring, the degree of unsaturation of the non-aromatic ring may be limited. Cycloalkyl groups may be optionally substituted with one or more substituents. In one embodiment, zero, one, two, three, or four atoms of each ring of the cycloalkyl group may be substituted with substituents. Typical examples of cycloalkyl groups include cyclopropyl, cyclopentyl, cyclohexyl, cyclobutyl, cycloheptyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, and cyclohexadienyl.

[0130] The term "cycloalkoxy" refers to an -O-cycloalkyl substituent.

[0131] The term "cycloalkoxyalkyl" refers to an alkyl-O-cycloalkyl substituent.

[0132] The term "cycloalkylalkoxy" refers to an -O-alkyl-cycloalkyl substituent.

[0133] The term "cycloalkylaminocarbonyl" refers to the -C(O)-NH-cycloalkyl substituent.

[0134] The term "aryl" refers to monocyclic, bicyclic, or tricyclic hydrocarbon aromatic ring systems. The aryl group may be optionally substituted with one or more substituents. In one embodiment, zero, one, two, three, four, five, or six atoms in each ring of the aryl group may be substituted with substituents. Examples of aryl groups include phenyl, naphthyl, anthracenyl, fluorenyl, indenyl, and azlenyl.

[0135] The term "aryloxy" refers to an -O-aryl substituent.

[0136] The term "arylalkoxy" refers to an -O-alkyl-aryl substituent.

[0137] The term "arylalkylthio" refers to an -S-alkyl-aryl substituent.

[0138] The term "arylthioalkyl" refers to an alkyl-S-aryl substituent.

[0139] The term "arylalkylaminocarbonyl" refers to a -C(O)-amino-alkyl-aryl substituent.

[0140] The term "arylalkylsulfonyl" refers to the -S(O)2-alkyl-aryl substituent.

[0141] The term "arylalkylsulfinyl" refers to the -S(O)-alkyl-aryl substituent.

[0142] The term "aryloxyalkyl" refers to an alkyl-O-aryl substituent.

[0143] The term "alkylaryl" refers to an aryl-alkyl substituent.

[0144] The term "arylalkyl" refers to an alkyl-aryl substituent.

[0145] The term "heteroaryl" refers to an aromatic 5- to 8-membered monocyclic, 8- to 12-membered bicyclic, or 11- to 14-membered tricyclic (having 1- to 4 ring heteroatoms in the case of monocyclics, 1- to 6 heteroatoms in the case of bicyclics, or 1- to 9 heteroatoms in the case of tricyclics), wherein the heteroatoms are selected from O, N, or S, and the remaining ring atoms are carbon (having appropriate hydrogen atoms unless otherwise indicated). The heteroaryl group may be optionally substituted with one or more substituents. In one embodiment, 0, 1, 2, 3, or 4 substituents may be substituted on each ring of the heteroaryl group. The heteroaryl group may be fully unsaturated, or it may be partially unsaturated and partially saturated. Examples of heteroaryl groups include pyridyl, furanyl, thienyl, pyrrolyl, oxazolyl, oxadiazolyl, imidazolyl, thiazolyl, isoxazolyl, quinolinyl, pyrazolyl, isothiazolyl, pyridadinyl, pyrimidinyl, pyrazinyl, triazinyl, isoquinolinyl, and indazolyl.

[0146] The term "heteroarylalkyl" refers to an alkyl-heteroaryl substituent.

[0147] The term "heteroaryloxy" refers to an -O-heteroaryl substituent.

[0148] The term "heteroarylalkoxy" refers to an -O-alkyl-heteroaryl substituent.

[0149] The term "heteroaryloxyalkyl" refers to an alkyl-O-heteroaryl substituent.

[0150] The term "nitrogen-containing heteroaryl" refers to a heteroaryl group that has 1 to 4 ring nitrogen heteroatoms in the case of a monocyclic group, 1 to 6 ring nitrogen heteroatoms in the case of a bicyclic group, or 1 to 9 ring nitrogen heteroatoms in the case of a tricyclic group.

[0151] The term "heterocycloalkyl" refers to a non-aromatic 3-8 member monocyclic system, a 7-12 member bicyclic system, or a 10-14 member tricyclic system (containing 1-3 heteroatoms in the case of a monocyclic system, 1-6 heteroatoms in the case of a bicyclic system, or 1-9 heteroatoms in the case of a tricyclic system), wherein the heteroatoms are selected from O, N, S, B, P, or Si, and the non-aromatic ring system is fully saturated. The heterocycloalkyl group may be optionally substituted with one or more substituents. In one embodiment, 0, 1, 2, 3, or 4 substituents may be substituted on each ring of the heterocycloalkyl group. Representative heterocycloalkyl groups include piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, 1,3-dioxolane, tetrahydrofuranyl, tetrahydrothienyl, and thirenyl.

[0152] The term "heterocycloalkylalkyl" refers to an alkyl-heterocycloalkyl substituent.

[0153] The term "alkylamino" refers to an amino substituent that is further substituted with one or two alkyl groups. The term "aminoalkyl" refers to an alkyl substituent that is further substituted with one or more amino groups. The term "hydroxyalkyl" or "hydroxylalkyl" refers to an alkyl substituent that is further substituted with one or more hydroxyl groups. The alkyl or aryl portion of alkylamino, aminoalkyl, mercaptoalkyl, hydroxyalkyl, mercaptoalkoxy, sulfonylalkyl, sulfonylaryl, alkylcarbonyl, and alkylcarbonylalkyl may optionally be substituted with one or more substituents.

[0154] [ka] The symbol indicates a bond point between a chemical structural part and the rest of the molecule or chemical formula.

[0155] The term "nucleic acid base" refers to nitrogen-containing biocompounds that form nucleosides. Examples of nucleic acid bases include purine bases and pyrimidine bases. The five nucleic acid bases adenine (A), cytosine (C), guanine (G), thymine (T), and uracil (U) are called the major or canonical nucleic acid bases. When nucleic acid bases are listed in a formula definition, the nucleic acid base refers to the part of the formula that is covalently bonded to it.

[0156] The term "modified nucleic acid base" refers to a derivative of a nucleic acid base. Examples of modified nucleic acid bases include, but are not limited to, xanthine, hypoxanthine, 7-methylguanine, 5,6-dihydrouracil, 5-methylcytosine, 5-hydroxymethylcytosine, purine, 2,6-diaminopurine, and 6,8-diaminopurine. When modified nucleic acid bases are listed in a formula definition, the modified nucleic acid base refers to the portion covalently bonded to the given formula.

[0157] The term “substituent” means an atom or group that replaces an atom or group of a given parent compound. For example, a substituent of a modified nucleoside is an atom or group different from the atom or group found in the natural nucleoside (for example, a modified 2'-substituent is an atom or group at the 2' position of the nucleoside that is any atom or group other than H or OH). Substituents may or may not be protected. Substituents may be further substituted with other substituents and may be bonded to the parent compound directly or via linking groups, such as alkyl or hydrocarbyl groups. Similarly, as used herein, “substituent” means, with respect to chemical functional groups, an atom or group different from the atom or group of atoms normally present in the given functional group. In certain embodiments, the substituents of any group (e.g., alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, heterocycloalkyl, etc.) can be on any of the atoms of that group, and any of the substituted groups (e.g., alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, heterocycloalkyl, etc.) can be optionally substituted with one or more substituents (which may be the same or different), each replacing a hydrogen atom.Examples of suitable substituents include alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano, nitro, alkoxy, aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl), carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl, alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy, heteroaryloxycarbonyl, thio, mercapto, mercaptoalkyl, arylsulfonyl, amino, aminoalkyl, dialkylamino, alkylcarbonylamino, aryl Examples include, but are not limited to, hydroxyaminocarbonyl, alkoxycarbonylamino, alkylamino, arylamino, diarylamino, aryl, arylalkylamino, aralkylaminocarbonyl, amide, alkylaminosulfonyl, arylaminosulfonyl, dialkylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, imino, carboxamide, carbamide, carbamyl, thioureido, thiocyanato, sulfamide, sulfonylalkyl, sulfonylaryl, mercaptoalkoxy, N-hydroxyamidinyl, N'-aryl, or N''-hydroxyamidinyl.In certain embodiments, substituents on any of the groups include alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano, nitro, alkoxy, aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl), carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl, alkylcarbonyloxy, thiocarbonyl, thio, mercapto, mercaptoalkyl, arylsulfonyl, amino, aminoalkyl, dialkylamino, alkylcarbonylamino, alkylaminocarbonyl, alkoxycarbonylamino, alkylamino, arylamino, diarylamino, aryl, arylalkylamino, aralkylaminocarbonyl, or amide substituted with alkylcarbonyl or arylamino. In certain embodiments, substituents on any of the groups include alkyl, halogen, haloalkyl, cyano, nitro, alkoxy, hydroxyl, hydroxylalkyl, carboxyl, formyl, alkylcarbonyl, alkoxycarbonyl, alkylcarbonyloxy, thio, mercapto, mercaptoalkyl, amino, aminoalkyl, dialkylamino, alkylcarbonylamino, alkylaminocarbonyl, or alkylamino.

[0158] The terms “protecting group” or “protecting moiety” refer to substituents commonly used to block or protect a particular function of other functional groups in a compound, its derivative, or its conjugate during reaction, and include nitrogen protecting groups when bonded to a nitrogen atom, or oxygen protecting groups when bonded to an oxygen atom. Nitrogen and oxygen protecting groups are well known in the art and include those described in detail in *Protecting Groups in Organic Synthesis*, TW Greene and PGMWuts, 3rd edition, John Wiley & Sons, 1999, which is incorporated herein by reference.

[0159] In certain embodiments, the substituent present on the nitrogen atom is a nitrogen protecting group (also referred to as an amino protecting group). Examples of nitrogen protecting groups include -OH, -OR aa , -N(R cc )2, -C(=O)R aa , -C(=O)N(R cc )2, -CO2R aa , -SO2R aa , -C(=NR cc )R aa , -C(=NR cc )OR aa , -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc , -SO2OR cc , -SOR aa , -C(=S)N(R cc )2, -C(=O)SR cc , -C(=S)SR cc , C 1-10 alkyl (e.g., aralkyl, heteroaralkyl), C 2-10 alkenyl, C 2-10 alkynyl, C 3-10 carbocyclic, 3- to 14-membered heterocyclic, C 6-14 aryl and 5- to 14-membered heteroaryl groups, among others, and are not limited thereto. Each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aralkyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R dd groups, and each R aa , R bb and R cc are independently alkyl, cycloalkyl, aryl, or heteroaryl, each of which may be optionally substituted with 1 to 3 independent R dd groups, and each R ddThese are independently alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano, nitro, alkoxy, aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl), carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl, alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy, heteroaryloxycarbonyl, thio, mercapto, mercaptoalkyl, arylsulfonyl, amino, aminoalkyl These include dialkylamino, alkylcarbonylamino, alkylaminocarbonyl, alkoxycarbonylamino, alkylamino, arylamino, diarylamino, aryl, arylalkylamino, aralkylaminocarbonyl, amide, alkylaminosulfonyl, arylaminosulfonyl, dialkylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, imino, carbamide, carbamyl, thioureido, thiocyanato, sulfamide, sulfonylalkyl, sulfonylaryl, or mercaptoalkoxy. Nitrogen protecting groups are well known in the art, as seen in Protecting Groups in Organic Synthesis, TW Greene and PGMWuts, 3 rd This includes the basis described in detail in edition, John Wiley & Sons, 1999, the references of which are incorporated herein by reference.

[0160] Amide nitrogen protecting group (e.g., -C(=O)R) aaExamples of these derivatives include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolineamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetacetamide, (N'-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazofenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine, o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

[0161] Carbamate nitrogen protecting group (e.g., -C(=O)OR) aaExamples include methyl carbamate, ethyl carbamate, 9-fluorenylmethylcarbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluoroenylmethylcarbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxantyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacylcarbamate (Phenoc), 2,2,2-trichloroethylcarbamate (Troc), and 2-trimethylsilyl Ethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bu meoc), 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamide)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyl Ludithiocarbamate, benzylcarbamate (Cbz), p-methoxybenzylcarbamate (Moz), p-nitobenzylcarbamate, p-bromobenzylcarbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzylcarbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethylcarbamate, diphenylmethylcarbamate, 2-methylthioethylcarbamate, 2-methylsulfonylethylcarbamate, 2-(p-toluenesulfonyl)ethylcarbamate, [2-(1,3-Dithianyl)methylcarbamate (Dmoc), 4-methylthiophenylcarbamate (Mtpc), 2,4-dimethylthiophenylcarbamate (Bmpc), 2-phosphonioethylcarbamate (Peoc), 2-triphenylphosphonioisopropylcarbamate (Ppoc), 1,1-dimethyl-2-cyanoethylcarbamate, m-chloro-p-acyloxybenzylcarbamate, p-(dihydroxyboryl)benzylcarbamate, 5-benzisoxazolylmethylcarbamate, 2-(trifluoromethyl)-6- Romonylmethylcarbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzylcarbamate, o-nitrobenzylcarbamate, 3,4-dimethoxy-6-nitrobenzylcarbamate, phenyl(o-nitrophenyl)methylcarbamate, t-amylcarbamate, S-benzylthiocarbamate, p-cyanobenzylcarbamate, cyclobutylcarbamate, cyclohexylcarbamate, cyclopentylcarbamate, cyanopropylmethylcarbamate, p-decyloxybezylcarbamate, 2, 2-Dimethoxyacyl vinylcarbamate, o-(N,N-dimethylcarboxamide)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamide)propylcarbamate, 1,1-dimethylpropynylcarbamate, di(2-pyridyl)methylcarbamate, 2-furanylmethylcarbamate, 2-iodoethylcarbamate, isobornylcarbamate, isobutylcarbamate, isonicotinylcarbamate, p-(p'-methoxyphenylazo)benzylcarbamate, 1-methylchlorobutylcarbamate, 1-Methylchlorohexylcarbamate, 1-Methyl-1-Cyanopropylmethylcarbamate, 1-Methyl-1-(3,5-Dimethoxyphenyl)ethylcarbamate, 1-Methyl-1-(p-Phenylazophenyl)ethylcarbamate, 1-Methyl-1-Phenylethylcarbamate, 1-Methyl-1-(4-Pyridyl)ethylcarbamate, Phenylcarbamate, p-(Phenylazo)benzylcarbamate, 2,4,6-Tri-t-butylphenylcarbamate, 4-(Trimethylammonium)benzylcarbamate and 2,4,Examples include, but are not limited to, 6-trimethylbenzylcarbamate.

[0162] Sulfonamide nitrogen protecting group (e.g., -S(=O)2R) aa Examples include p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6 Examples include, but are not limited to, dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

[0163] Other nitrogen protecting groups include phenothiazinyl-(10)-acyl derivatives, N'-p-toluenesulfonylaminoacyl derivatives, N'-phenylaminothioacyl derivatives, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, and N-1,1,4,4-tetramethyldisilylazacyl Clopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexane-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexane-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrroline-3-yl) Mine, quaternary ammonium salt, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthio Methyleneamine, N-benzylideneamine, Np-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N-(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, Np-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexyllideneamine, N-(5,Examples include, but are not limited to, 5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivatives, N-diphenylboric acid derivatives, N-[phenyl(pentaacylchromium or pentaacyltungsten)acyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkylphosphoramide, dibenzylphosphoramide, diphenylphosphoramide, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

[0164] In certain embodiments, the substituent present on the oxygen atom is an oxygen protecting group (also called a hydroxyl protecting group). Examples of oxygen protecting groups include -R aa , -N(R bb )2, -C(=O)SR aa -C(=O)R aa , -CO2R aa -C(=O)N(R bb )2, -C(=NR bb )R aa -C(=NR bb )OR aa -C(=NR bb )N(R bb )2, -S(=O)R aa , -SO2R aa , -Si(R aa ) 3、 -P(R cc )2, -P(R cc )3, -P(=O)2R aa -P(=O)(R aa )2, -P(=O)(OR cc )2, -P(=O)2N(R bb )2 and -P(=O)(NR bb )2 is one example, but it is not limited to these, and in the formula, Raa , R bb and R cc The oxygen protecting group is as defined herein. The oxygen protecting group is well known in the art, as seen in Protecting Groups in Organic Synthesis, TWGreene and PGMWuts, 3 rd This includes the basis described in detail in edition, John Wiley & Sons, 1999, the references of which are incorporated herein by reference.

[0165] Examples of oxygen protecting groups include methyl, methoxymethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-Trichloroethoxymethyl, Bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), Tetrahydropyranyl (THP), 3-Bromotetrahydropyranyl, Tetrahydrothiopyranyl, 1-Methoxycyclohexyl, 4-Methoxytetrahydropyranyl (MTHP), 4-Methoxytetrahydrothiopyranyl, 4-Methoxytetrahydrothiopyranyl S,S-Dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methyl Toxypiperidine-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluorooloethyl, 2,2,2-trichloroethyl Tyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxide, diphenylmethyl, p,p'-Dinitrobenzhydryl, 5-Dibenzosberyl, Triphenylmethyl, α-Naphthyldiphenylmethyl, p-Methoxyphenyldiphenylmethyl, Di(p-Methoxyphenyl)phenylmethyl, Tri(p-Methoxyphenyl)methyl, 4-(4'-Bromophenacyloxyphenyl)diphenylmethyl, 4,4',4”-Tris(4,5-Dichlorophthalimidophenyl)methyl, 4,4',4”-Tris(Lebrinoyloxyphenyl)methyl, 4,4',4”-Tris(Benzoyloxyphenyl)methyl, 3-(I Midazole-1-yl)bis(4',4"-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxide, trimethylsilyl (TMS), trimethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), Dimethyltexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, bensoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chloropheno Xyacetate, 3-phenylpropionate, 4-oxopentanoate (rebrinate), 4,4-(ethylenedithio)pentanoate (rebrinoyldithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), t-butyl carbonate (BOC), alkylmethyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkylethyl carbonate, alkyl 2,22-Trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonio)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzylthiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenes Sulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinate, (E)-2-methyl Examples include, but are not limited to, tyl-2-butenoate, o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphodiamide, alkyl N-phenylcarbamate, borate, dimethylphosphinthiole, alkyl 2,4-dinitrophenyl sulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts).

[0166] In certain embodiments, the substituent present on the sulfur atom is a sulfur protecting group (also called a thiol protecting group). Examples of sulfur protecting groups include -R aa , -N(R bb )2, -C(=O)SR aa -C(=O)R aa , -CO2R aa-C(=O)N(R bb )2, -C(=NR bb )R aa -C(=NR bb )OR aa -C(=NR bb )N(R bb )2, -S(=O)R aa , -SO2R aa , -Si(R aa ) 3、 -P(R cc )2, -P(R cc )3, -P(=O)2R aa -P(=O)(R aa )2, -P(=O)(OR cc )2, -P(=O)2N(R bb )2 and -P(=O)(NR bb )2 is one example, but it is not limited to these, and in the formula, R aa , R bb and R cc The sulfur protecting group is as defined herein. The sulfur protecting group is well known in the art, as seen in Protecting Groups in Organic Synthesis, TWGreene and PGMWuts, 3 rd This includes the basis described in detail in edition, John Wiley & Sons, 1999, the references of which are incorporated herein by reference.

[0167] The terms "antisense oligonucleotide" or "antisense chain" refer to an oligonucleotide that contains a region complementary to the target nucleic acid.

[0168] The terms “composition” or “pharmaceutical composition” mean a mixture of substances suitable for administration to a subject. For example, a composition may contain one or more compounds or their salts and sterile aqueous solutions.

[0169] The term "nucleic acid" refers to a molecule composed of linked monomeric nucleotides or nucleosides. Examples of nucleic acids include, but are not limited to, ribonucleic acid (RNA), deoxyribonucleic acid (DNA), single-stranded nucleic acids, and double-stranded nucleic acids.

[0170] The term "nucleic acid base sequence" refers to the sequence of nucleic acid bases in a nucleic acid or oligonucleotide, independently of any sugar or nucleoside bond.

[0171] The term "nucleoside" refers to a compound containing a nucleic acid base and a sugar moiety. The nucleic acid base and sugar moiety can be independently either unmodified or modified. A "modified nucleoside" refers to a nucleoside containing a modified nucleic acid base and / or a modified sugar moiety. Modified nucleosides include debasic nucleosides, which lack a nucleic acid base.

[0172] The term "oligomeric compound" refers to a polymer in which subunits are linked together. With respect to proteins, peptides, polypeptides, or antibodies, "subunit" refers to an amino acid bond or a peptide bond. With respect to oligonucleotides, "subunit" refers to a nucleotide, nucleoside, nucleic acid base, or sugar, or a modified nucleotide, modified nucleoside, modified nucleic acid base, or modified sugar, as provided in this invention.

[0173] The term “oligonucleotide” means a polymer of linked nucleosides (e.g., polynucleotides, nucleic acids, polymers of nucleosides), each of which may be modified or unmodified independently of the others. Oligonucleotides may consist of, but are not limited to, ribonucleic acids (e.g., composed of ribonucleosides), deoxyribonucleic acids (e.g., composed of deoxyribonucleosides), modified nucleic acids (e.g., composed of modified nucleic acid bases, modified sugars and / or modified phosphate groups), or combinations thereof. Examples of oligonucleotide compounds include single-stranded and double-stranded compounds, e.g., oligonucleotides, antisense oligonucleotides, interfering RNA compounds (RNAi compounds), oligonucleotide-targeting microRNAs (miRNAs), miRNA mimes, occupation-based compounds (e.g., compounds that block mRNA processing or translation, and splicing compounds), and editing compounds (e.g., ADAR recruiting molecules, ADAR targeting molecules, single-stranded guide nucleic acids, or combinations thereof). RNAi compounds include double-stranded compounds (e.g., small interfering RNA (siRNA) and double-stranded RNA (dsRNA)) and single-stranded compounds (e.g., single-stranded siRNA (ssRNA), single-stranded RNAi (ssRNAi), short hairpin RNA (shRNA), and microRNA mimetic compounds), which act at least partially via the RNA-induced silencing complex (RISC) pathway, resulting in sequence-specific degradation and / or sequestration of target nucleic acids through a process known as RNA interference (RNAi). The term “RNAi compound” is intended to be equivalent to other terms used to describe nucleic acid compounds that can mediate sequence-specific RNA interference, such as interfering RNA (iRNA), iRNA agents, RNAi agents, small interfering RNA, small interfering RNA, small interfering oligonucleotides, small interfering nucleic acids, small interfering modified oligonucleotides, and chemically modified siRNA. In addition, the term “RNAi” is intended to be equivalent to other terms used to describe sequence-specific RNA interference.

[0174] The terms “target nucleic acid,” “target RNA,” and “nucleic acid target” all refer to nucleic acids that can lead to the compounds described herein.

[0175] The term "therapeutic compound" includes any pharmaceutical agent or pharmaceutical compound that provides a therapeutic benefit. Examples of therapeutic compounds include nucleic acids, oligomeric compounds, oligonucleotides, proteins, peptides, antibodies, small molecules, and other such agents.

[0176] The "target region" refers to the portion of a target nucleic acid into which one or more compounds are directed.

[0177] The "targeting portion" refers to a conjugate group that increases the affinity for a given target, such as a molecule, cell or cell type, compartment, such as a compartment of a cell or organ, tissue, organ or region, compared to a compound in which such portion does not exist.

[0178] "Terminal group" refers to the chemical group or atomic group covalently bonded to the end of an oligonucleotide.

[0179] The term "ligand" refers to a substance that binds to or interacts with proteins, nucleic acids, or other biomolecules. In some embodiments, the ligand is a small molecule. In some embodiments, the ligand binds to a protein (e.g., a receptor). In certain embodiments, the ligand binds to an NMDA receptor.

[0180] The term "conjugated" refers to two molecules that are joined together by a covalent bond (for example, an oligonucleotide and a ligand as described herein). For example, a covalent bond to a ligand or oligonucleotide can be formed at either an atom of that ligand or oligonucleotide (i.e., a radical of that ligand or oligonucleotide).

[0181] The term "N-methyl-D-aspartate receptor" or "NMDA receptor" refers to a glutamate receptor and ion channel found, for example, in human neurons. Many ligands for NMDA receptors are known in the art and are disclosed, for example, in Neuropharmacology 2007, 53(6), 699-723, J.Med.Chem. 1990, 33(2), 789-808, Neuroscience 2001, 105(3), 663-669, J.Med.Chem. 2022, 65(13), 9063-9075, Drugs Fut. 2004, 29(10), 992, Drugs Fut. 2004, 29(10), 993, and British Journal of Pharmacology 2022, 179(6), 1146-1187, each of which is incorporated herein by reference.

[0182] The term "sense oligonucleotide" or "sense chain" refers to a chain of a double-stranded compound that contains a region substantially complementary to the antisense chain of that double-stranded compound.

[0183] The terms “microRNA” and “miRNA” may be used synonymously herein and refer to short (e.g., about 20–24 nucleosides long) non-coding ribonucleic acid (RNA) that is involved in the post-transcriptional regulation of gene expression in multicellular organisms by affecting both mRNA stability and translation. miRNA is transcribed as part of a primary transcript (pri-miRNA) that is capped and polyadenylated by RNA polymerase II and can be either protein-coding or non-coding. This primary transcript is cleaved by the enzyme Drosha ribonuclease III to produce a stem-loop precursor miRNA (pre-miRNA) about 70 nucleosides long, which is further processed in the RNAi pathway. As part of this pathway, the pre-miRNA is cleaved by cytoplasmic dicer ribonuclease to produce mature miRNA and antisense miRNA star (miRNA*) products. The mature miRNA is incorporated into the RNA-induced silencing complex (RISC), which recognizes the target mRNA through incomplete base pairing (i.e., partial complementarity) with the miRNA, most commonly resulting in the inhibition or destabilization of the translation of the target mRNA. This mechanism is most often seen through the fact that the miRNA can reduce gene expression by either binding to the 3' untranslated region (UTR) of the target mRNA and inhibiting translation (e.g., by blocking ribosomes from reaching it for translation) or by directly degrading the transcript. The term (i.e., miRNA) may be used herein for the purpose of referring to any form of target miRNA (e.g., precursor miRNA, primary miRNA, and / or mature miRNA).

[0184] The terms “small interfering RNA,” “low interfering RNA,” and “siRNA” may be used synonymously herein and refer to RNA molecules that are useful in RNA interference (RNAi) and may exist as non-coding double-stranded RNA (dsRNA) molecules with a length of about 20 to about 24 nucleosides. siRNA is often found phosphorylated at the 5' end and hydroxylated at the 3' end, with its 3' end typically having an overhang of two nucleotides beyond the 5' end of its antiparallel strand (e.g., the complementary strand of its dsRNA molecule). siRNA can prevent translation (e.g., inhibit, silence, or interfere) by binding to a complementary target sequence (e.g., a target nucleic acid sequence) and thereby inhibiting the expression of a specific gene by promoting (e.g., promoting, inducing, or initiating) the degradation of that mRNA. After integrating and separating to form a RISC complex, siRNA prevents its mRNA from being used as a translation template by forming base pairs with its target mRNA (e.g., complete complementarity) and cleaving it. As described above in this specification, the RISC complex incorporating miRNA is also part of the RNAi pathway, but it scans cytoplasmic mRNA for potential complementarity (e.g., partial complementarity).

[0185] The term “ADAR recruiting molecule” refers to a nucleic acid configured to increase the concentration of the ribonucleic acid-acting adenosine deaminase (ADAR) enzyme at a local site around that nucleic acid, as may be used herein. In some embodiments, the increase in concentration is relative to the concentration at a given local site where the ADAR recruiting molecule is not present. In some embodiments, the ADAR recruiting molecule includes double-stranded RNA.

[0186] The term “ADAR targeting molecule” refers to a nucleic acid configured to guide an ADAR molecule to a desired location (e.g., local). As used herein, the term “guide” means increasing the concentration of ADAR at the desired location compared to the concentration in the absence of the ADAR targeting molecule. In some embodiments, the ADAR targeting molecule can be configured to control the desired location by altering the sequence and / or properties of the nucleic acid (e.g., by modification of nucleic acid bases, sugars, nucleoside bonds, or other components). In some embodiments, the ADAR targeting molecule comprises an ADAR recruiting molecule and a single-stranded guide nucleic acid. In some embodiments, the ADAR targeting molecule comprises a double-stranded RNA and a single-stranded guide nucleic acid.

[0187] The terms “single-stranded guide nucleic acid” or “guide RNA” refer to a single-stranded nucleic acid containing a specific sequence that is at least partially complementary to the target sequence, as may be used herein. In some embodiments, the target sequence is located locally, adjacent to, or near the desired site for regulating ADAR concentration. In some embodiments, the level of complementarity is sufficient to facilitate the binding (e.g., annealing) of the single-stranded guide nucleic acid to the target sequence.

[0188] The compounds of this disclosure may also contain one or more atomic isotopes in non-natural proportions of the atoms constituting such compounds. For example, the compounds may contain radioactive isotopes, such as tritium. 3 H), Iodine-125( 125 I) or carbon-14 ( 14 C) may be radioactively labeled. All isotopic variants of the compounds disclosed herein, whether radioactive or not, are included within the scope of this disclosure.

[0189] The term “isotope variant” refers to a therapeutic agent (e.g., compounds and / or modified oligonucleotides disclosed herein) that contains one or more isotopes in non-natural proportions of the atoms constituting such therapeutic agents. In certain embodiments, the “isotope variant” of a therapeutic agent contains one or more isotopes in non-natural proportions, such as hydrogen (H), deuterium ( 2 H), tritium ( 3 H), carbon-11 ( 11 C), carbon-12( 12 C), carbon-13( 13 C), carbon-14( 14 C), nitrogen-13( 13 N), Nitrogen-14( 14 N), Nitrogen-15( 15 N), oxygen-14( 14 O), oxygen-15( 15 O), oxygen-16( 16 O), oxygen-17( 17 O), oxygen-18( 18 O), Fluorine-17( 17 F), Fluorine-18( 18 F), phosphorus-31( 31 P), phosphorus-32( 32 P), Lin-33( 33 P), sulfur-32( 32 S), sulfur-33( 33 S), Sulfur-34( 34 S), sulfur-35( 35 S), sulfur-36( 36 S), Chlorine-35( 35 Cl), Chlorine-36( 36 Cl), Chlorine-37( 37 Cl), bromine-79( 79 Br), Bromine-81( 81 Br), Iodine-123 ( 123 I) Iodine-125 125 I) Iodine-127( 127 I) Iodine-129( 129 I) and Iodine-131 131 I) is one example, but is not limited to these. In certain embodiments, the “isotope variant” of the therapeutic agent contains one or more isotopes in non-natural proportions, such as hydrogen (H), deuterium (2 H), tritium ( 3 H), carbon-11 ( 11 C), carbon-12( 12 C), carbon-13( 13 C), carbon-14( 14 C), nitrogen-13( 13 N), Nitrogen-14( 14 N), Nitrogen-15( 15 N), oxygen-14( 14 O), oxygen-15( 15 O), oxygen-16( 16 O), oxygen-17( 17 O), oxygen-18( 18 O), Fluorine-17( 17 F), Fluorine-18( 18 F), phosphorus-31( 31 P), phosphorus-32( 32 P), Lin-33( 33 P), sulfur-32( 32 S), sulfur-33( 33 S), Sulfur-34( 34 S), sulfur-35( 35 S), sulfur-36( 36 S), Chlorine-35( 35 Cl), Chlorine-36( 36 Cl), Chlorine-37( 37 Cl), bromine-79( 79 Br), Bromine-81( 81 Br), Iodine-123 ( 123 I) Iodine-125 125 I) Iodine-127( 127 I) Iodine-129( 129 I) and Iodine-131 131 I) is one example, but it is not limited to these.

[0190] In therapeutic agents (e.g., compounds and / or modified oligonucleotides disclosed herein), where feasible according to the judgment of those skilled in the art, for example, any hydrogen 2 It can be H, and for example, any carbon 13 It can be C, and for example, any nitrogen 15 It can be N, or for example, any oxygen18 It should be understood that it can be O. In certain embodiments, the “isotope variant” of the therapeutic agent contains a non-natural proportion of deuterium (D).

[0191] A "modified oligonucleotide" refers to an oligonucleotide in which at least one sugar, nucleic acid base, or nucleoside bond has been modified.

[0192] "Nucleic acid base sequence" refers to the sequence of consecutive nucleic acid bases in a nucleic acid or oligonucleotide, independent of any sugar or nucleoside bonds.

[0193] The term "oligomeric double-stranded" refers to a double-stranded structure formed by two oligomeric compounds having complementary nucleic acid base sequences. Each oligomeric compound in an oligomeric double-stranded structure is sometimes referred to as a "double-stranded oligomeric compound." Each oligomeric compound in an oligomeric double-stranded structure may contain a non-complementary overhang nucleoside. In some embodiments, the terms "double-stranded oligomeric compound" and "modified oligonucleotide" are used synonymously. In other embodiments, the terms "oligomeric double-stranded structure" and "compound" are used synonymously.

[0194] A "phosphorothioate bond" refers to a modified phosphate bond in which one of the non-bridged oxygen atoms is replaced by a sulfur atom.

[0195] The terms “RNA interference compound,” “RNAi compound,” and / or “iRNA agent” refer to compounds that act, at least in part, via the RNA-induced silencing complex (RISC) pathway or Ago2, rather than via RNase H, to modulate target nucleic acids and / or target proteins encoded by the target nucleic acid. RNAi compounds include, but are not limited to, double-stranded siRNA, single-stranded siRNA, and microRNA (including microRNA mimetic).

[0196] Specific Embodiments In certain embodiments, the compound comprises an NMDA receptor ligand and one or more linker moieties. In certain embodiments, the compound is selected from any of the formulas I, II, II-a, III, III-a, IV, IV-a, V, Va, VI, VI-a, VI-b, VII, VII-a, VIII, VIII-a, IX, IX-a, X, Xa or salts thereof, as described herein. In certain embodiments, one or more linker moieties (e.g., L1, L2, L3, L4) link the NMDA receptor ligand to a therapeutic, prophylactic, or diagnostic agent. In certain embodiments, the compound further comprises one or more therapeutic, prophylactic, or diagnostic agents. In certain embodiments, the therapeutic, prophylactic, or diagnostic agent is a small molecule or an oligomeric compound. In certain embodiments, the oligomeric compound comprises a protein, peptide, antibody, oligonucleotide, or a combination thereof.

[0197] In certain embodiments, the oligomeric compound is one of those described herein. In certain embodiments, the oligomeric compound has a subunit length of about 10 to 50. In certain embodiments, the oligomeric compound is an oligonucleotide. In certain embodiments, the oligonucleotide is one of those described herein. In certain embodiments, the oligonucleotide has a linked nucleoside length of 8 to 80, or a linked nucleoside length of 12 to 50, or a linked nucleoside length of 12 to 30, or a linked nucleoside length of 15 to 30.

[0198] In certain embodiments, the oligonucleotide is a modified oligonucleotide comprising at least one modified nucleoside bond, at least one modified sugar, or at least one modified nucleic acid base.

[0199] In certain embodiments, the oligonucleotide is single-stranded. In certain embodiments, the oligonucleotide is double-stranded. In certain embodiments, the oligonucleotide is double-stranded over a portion of its length. In certain embodiments, the oligonucleotide includes ribonucleic acid (e.g., composed of ribonucleosides), deoxyribonucleic acid (e.g., composed of deoxyribonucleosides), or a combination thereof. In certain embodiments, the oligonucleotide is small interfering RNA (siRNA), microRNA (miRNA) antagonists, miRNA mimetic molecules, ADAR recruiting molecules, ADAR targeting molecules, guide RNA, antisense oligonucleotides, short hairpin RNA (shRNA), or a combination thereof.

[0200] In some embodiments, the linker is an optionally substituted alkyl linker. In some embodiments, the linker is an optionally substituted C1-C6 alkyl linker. In some embodiments, the linker is a C1-C6 alkyl linker substituted with =O. In certain embodiments, the linker is [ka] The structure includes the following: In some embodiments, the linker is optionally replaced with C1-C 15 It is an alkyl linker. In some embodiments, the linker is optionally substituted C5-C 12 It is an alkyl linker. In certain embodiments, the linker is [ka] The structure includes, [ka] It includes the structure.

[0201] In some embodiments, the linker is a bond. In some embodiments, the linker is an optionally substituted PEG linker. In some embodiments, the linker is an optionally substituted PEG linker, wherein the length of the PEG units is 1, 2, 3, 4, 5, 6, 7, or 8, and the PEG units are [ka] The structure includes. In certain embodiments, the linker is an optionally substituted PEG linker, where the length of the PEG unit is 3. In certain embodiments, the linker is an optionally substituted PEG linker, where the length of the PEG unit is 4. In certain embodiments, the linker is [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] It includes the structure.

[0202] In some embodiments, the linker is an optionally substituted heteroalkyl linker. In certain embodiments, the linker is [ka] The structure includes, [ka] It includes the structure.

[0203] In some embodiments, the linker is an optionally substituted heteroaryl linker. In some embodiments, the linker is an optionally substituted partially unsaturated heterocycloalkyl linker or heteroaryl linker. In some embodiments, the linker is [ka] It includes the structure.

[0204] In some embodiments, the linker is an optionally substituted heteroalkyl linker. In some embodiments, the linker is substituted with one or more =O substituents. In certain embodiments, the linker is [ka] The structure includes, where X is O or S. In certain embodiments, the linker is [ka] The structure includes the form where X is either O or S.

[0205] In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes, where X is O or S. In some embodiments, the linker is [ka] The structure includes the form where X is either O or S.

[0206] In certain embodiments, the compound is [ka] [ka] [ka] [ka] [ka] [ka] Or it contains or consists of one of the structures of these salts, where X is O or S.

[0207] In some embodiments, X is O. In some embodiments, X is S.

[0208] In some embodiments, R 1It contains an oligonucleotide. In some embodiments, the oligonucleotide is bonded at its 5' end. In some embodiments, the oligonucleotide is bonded at its 3' end. In some embodiments, the oligonucleotide is bonded at an internal position on the oligonucleotide. In some embodiments, the internal position is at an internucleoside bond. In some embodiments, R 1 This comprises an oligonucleotide conjugated with one or more additional NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated with two, three, four, or five or more additional NMDA receptor ligands. In certain embodiments, the additional NMDA receptor ligands are conjugated to the oligonucleotide at one or more of the 5' end, the 3' end, one or more internal positions on the oligonucleotide, or a combination thereof. In certain embodiments, the oligonucleotide is a modified oligonucleotide.

[0209] Certain embodiments provide compositions comprising a compound of any embodiment herein and a pharmaceutically acceptable carrier or excipient.

[0210] Certain embodiments provide compositions comprising compounds of any embodiment herein for use in therapy.

[0211] In certain embodiments, a method for delivering a drug to a cell includes delivering the drug to the cell by contacting the cell with a compound of any embodiment herein. In certain embodiments, the cell expresses an NMDA receptor on its surface. In certain embodiments, the cell is a brain cell. In certain embodiments, the cell is a cell of the prefrontal cortex. In certain embodiments, the cell is a cell of the striatum. In certain embodiments, the cell is a cell of the cerebellum. In certain embodiments, the cell is a cell of the brainstem. In certain embodiments, the cell is a cell of the hippocampus. In certain embodiments, the cell is a cell of the spinal cord. In certain embodiments, the drug is a therapeutic or diagnostic agent. In certain embodiments, the cell is located in an animal.

[0212] In certain embodiments, a method for regulating the expression of a nucleic acid target in a cell includes regulating the expression of the nucleic acid target in the cell by contacting the cell with a compound of any embodiment herein. In certain embodiments, the cell expresses an NMDA receptor on its surface. In certain embodiments, the cell is a brain cell. In certain embodiments, the cell is a cell of the prefrontal cortex. In certain embodiments, the cell is a cell of the striatum. In certain embodiments, the cell is a cell of the cerebellum. In certain embodiments, the cell is a cell of the brainstem. In certain embodiments, the cell is a cell of the hippocampus. In certain embodiments, the cell is a cell of the spinal cord. In certain embodiments, the agent is a therapeutic or diagnostic agent. In certain embodiments, contact between the cell and a compound of any embodiment herein inhibits the expression of the nucleic acid target. In certain embodiments, the nucleic acid target is a premRNA, mRNA, non-coding RNA, or miRNA. In certain embodiments, the cell is located in an animal.

[0213] In certain embodiments, a method for regulating the expression of a nucleic acid target in a subject includes regulating the expression of the nucleic acid target in the subject by administering one of the compounds or compositions provided in the present invention to the subject. In certain embodiments, the expression of the nucleic acid is regulated in cells of the subject that express an NMDA receptor on the surface of the cell. In certain embodiments, the expression of the nucleic acid is regulated in brain cells. In certain embodiments, the cells that express an NMDA receptor on the surface are brain cells. In certain embodiments, the brain cells are cells of the prefrontal cortex. In certain embodiments, the brain cells are cells of the striatum. In certain embodiments, the brain cells are cells of the cerebellum. In certain embodiments, the brain cells are cells of the brainstem. In certain embodiments, the brain cells are cells of the hippocampus. In certain embodiments, the brain cells are cells of the spinal cord. In certain embodiments, the nucleic acid target is a premRNA, mRNA, non-coding RNA, or miRNA. In certain embodiments, the compound is administered intrathecally to the subject.

[0214] In certain embodiments, a method for treating or improving a disease, disorder, or symptom in a subject includes treating, preventing, or improving the disease, disorder, or symptom in the subject by administering one of the compounds or compositions provided in the present invention to the subject. In certain embodiments, the disease, disorder, or symptom is a disease, disorder, or symptom of the central nervous system (CNS). In certain embodiments, the disease, disorder, or symptom is Alzheimer's disease or a symptom of Alzheimer's disease. In certain embodiments, the compound is administered intrathecally to the subject. In certain embodiments, the compound or composition is administered to the subject in a therapeutically effective dose.

[0215] In certain embodiments, compounds containing an NMDA receptor ligand selectively or preferentially target cells expressing the NMDA receptor compared to cells that do not express the NMDA receptor.

[0216] Furthermore, the present invention provides for the use of compounds such as those described herein in the manufacture of pharmaceuticals for the treatment of diseases or disorders.

[0217] In another aspect, the present disclosure provides a method for producing any of the compounds provided in the present invention, comprising one or more compounds and chemical conversions described herein (including Examples 1 to 77).

[0218] Certain compounds containing oligonucleotides In certain embodiments, the compounds described herein include oligonucleotides. In certain embodiments, the oligonucleotide has a nucleic acid base sequence that is at least partially complementary to a target nucleic acid sequence (e.g., a target nucleic acid expressed in an intracellular space). In some embodiments, the oligonucleotide can modify the expression of the original gene when delivered to a cell expressing the target nucleic acid. In some embodiments, the oligonucleotide can inhibit the expression of the original gene when delivered to a cell expressing the target nucleic acid. The gene expression can be modified or inhibited in vitro or in vivo. In certain embodiments, the oligonucleotide includes one or more ribonucleic acids (e.g., one or more ribonucleosides), deoxyribonucleic acids (e.g., one or more deoxyribonucleosides), modified nucleic acids (e.g., one or more modified nucleic acid bases, modified sugars, and / or modified nucleotide interbonds), or combinations thereof. In some embodiments, the oligonucleotide includes ribonucleic acid (RNA). In some embodiments, the oligonucleotide includes deoxyribonucleic acid (DNA). In some embodiments, the oligonucleotide includes modifications (e.g., modified nucleic acid bases, modified sugars, or modified nucleoside interbonds).

[0219] In certain embodiments, the oligonucleotide is single-stranded. In some embodiments, the single-stranded oligonucleotide is single-stranded RNA (ssRNA), ssDNA, or an ssRNA / DNA hybrid (e.g., a single-stranded oligonucleotide composed of both a ribonucleoside (modified or unmodified) and a deoxyribonucleoside (modified or unmodified)). In some embodiments, the oligonucleotide is double-stranded (e.g., composed of two single-stranded nucleic acids). Such a double-stranded oligonucleotide comprises a first oligonucleotide having a region complementary to the target nucleic acid and a second oligonucleotide having a region complementary to the first oligonucleotide. The first and second oligonucleotides may be independently modified. In certain embodiments, the first oligonucleotide is ligated to one or more NMDA receptor ligands. In certain embodiments, the second oligonucleotide is ligated to one or more NMDA receptor ligands.

[0220] In some embodiments, the oligonucleotide has a nucleoside length of at least 2 (for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35) ,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72,73,74,75,76,77,78,79,80 ,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97,98,99,100,101,102,103,104,105,106,107,108,109,110,111,112,113,114,115,116,117,118,1 The oligonucleotides are 19, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 or more. In some embodiments, the oligonucleotide has a length of at least 5 nucleosides. In some embodiments, the oligonucleotide has a length of at least 10 nucleosides. In some embodiments, the oligonucleotide has a length of at least 15 nucleosides. In some embodiments, the oligonucleotide has a length of at least 16 nucleosides. In some embodiments, the oligonucleotide has a length of at least 17 nucleosides. In some embodiments, the oligonucleotide has a length of at least 18 nucleosides. In some embodiments, the oligonucleotide has a length of at least 19 nucleosides. In some embodiments, the oligonucleotide has a length of at least 20 nucleosides. In some embodiments, the oligonucleotide has a length of at least 21 nucleosides. In some embodiments, the oligonucleotide has a length of at least 22 nucleosides. In some embodiments, the oligonucleotide has a length of at least 23 nucleosides.In some embodiments, the oligonucleotide has a length of at least 24 nucleosides. In some embodiments, the oligonucleotide has a length of at least 25 nucleosides. In some embodiments, the oligonucleotide has a length of at least 26 nucleosides. In some embodiments, the oligonucleotide has a length of at least 27 nucleosides. In some embodiments, the oligonucleotide has a length of at least 28 nucleosides. In some embodiments, the oligonucleotide has a length of at least 29 nucleosides. In some embodiments, the oligonucleotide has a length of at least 30 nucleosides. In some embodiments, the oligonucleotide has a length of at least 40 nucleosides. In some embodiments, the oligonucleotide has a length of at least 50 nucleosides. In some embodiments, the oligonucleotide has a length of at least 60 nucleosides. In some embodiments, the oligonucleotide has a length of at least 70 nucleosides. In some embodiments, the oligonucleotide has a length of at least 80 nucleosides. In some embodiments, the oligonucleotide has a length of at least 90 nucleosides. In some embodiments, the oligonucleotide has a length of at least 100 nucleosides. In some embodiments, the oligonucleotide is at least 150 nucleosides long.

[0221] In some embodiments, oligonucleotides have a nucleoside length of 150 or less (for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 3 5, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 11 8, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150). In some embodiments, the oligonucleotide has a nucleoside length of 150 or less. In some embodiments, the oligonucleotide has a nucleoside length of 100 or less. In some embodiments, the oligonucleotide has a nucleoside length of 90 or less. In some embodiments, the oligonucleotide has a nucleoside length of 80 or less. In some embodiments, the oligonucleotide has a nucleoside length of 70 or less. In some embodiments, the oligonucleotide has a nucleoside length of 60 or less. In some embodiments, the oligonucleotide has a nucleoside length of 50 or less. In some embodiments, the oligonucleotide has a nucleoside length of 40 or less. In some embodiments, the oligonucleotide has a nucleoside length of 30 or less. In some embodiments, the oligonucleotide has a nucleoside length of 29 or less. In some embodiments, the oligonucleotide has a nucleoside length of 28 or less. In some embodiments, the oligonucleotide has a nucleoside length of 27 or less.In some embodiments, the oligonucleotide has a nucleoside length of 26 or less. In some embodiments, the oligonucleotide has a nucleoside length of 25 or less. In some embodiments, the oligonucleotide has a nucleoside length of 24 or less. In some embodiments, the oligonucleotide has a nucleoside length of 23 or less. In some embodiments, the oligonucleotide has a nucleoside length of 22 or less. In some embodiments, the oligonucleotide has a nucleoside length of 21 or less. In some embodiments, the oligonucleotide has a nucleoside length of 20 or less. In some embodiments, the oligonucleotide has a nucleoside length of 19 or less. In some embodiments, the oligonucleotide has a nucleoside length of 18 or less. In some embodiments, the oligonucleotide has a nucleoside length of 17 or less. In some embodiments, the oligonucleotide has a nucleoside length of 16 or less. In some embodiments, the oligonucleotide has a nucleoside length of 15 or less. In some embodiments, the oligonucleotide has a nucleoside length of 10 or less. In some embodiments, the oligonucleotide has a nucleoside length of 5 or less.

[0222] In some embodiments, the oligonucleotide has a length of about 5 nucleosides to about 150 nucleosides. In some embodiments, the oligonucleotide has a length of about 10 nucleosides to about 100 nucleosides. In some embodiments, the oligonucleotide has a length of about 20 nucleosides to about 90 nucleosides. In some embodiments, the oligonucleotide has a length of about 30 nucleosides to about 80 nucleosides. In some embodiments, the oligonucleotide has a length of about 40 nucleosides to about 70 nucleosides. In some embodiments, the oligonucleotide has a length of about 50 nucleosides to about 60 nucleosides.

[0223] In some embodiments, the oligonucleotide is a therapeutic oligonucleotide. Therapeutic oligonucleotides may include, for example, small interfering RNA (siRNA), microRNA (miRNA) antagonists, miRNA mimetic molecules, ADAR recruiting molecules, ADAR targeting molecules, guide RNA, antisense oligonucleotides, short hairpin RNA (shRNA), or combinations thereof.

[0224] In certain embodiments, the miRNA is a precursor miRNA, a primary miRNA, and / or a mature miRNA.

[0225] In certain embodiments, the oligonucleotide comprises or consists of an antisense oligonucleotide. In certain embodiments, the antisense oligonucleotide is complementary to mRNA. In certain embodiments, the antisense oligonucleotide is complementary to premRNA. In certain embodiments, the antisense oligonucleotide blocks the translation of its mRNA transcript and promotes its degradation. In certain embodiments, the antisense oligonucleotide recruits RNase H and promotes the degradation of its mRNA transcript. In certain embodiments, the antisense oligonucleotide targets a miRNA to inhibit the miRNA from regulating mRNA expression and promote the degradation of the miRNA.

[0226] Specific modifications In certain embodiments, this disclosure relates to compounds comprising oligonucleotides. In certain embodiments, the oligonucleotide may be unmodified RNA or DNA, or it may be modified. In certain embodiments, the oligonucleotide is a modified oligonucleotide. In certain embodiments, the modified oligonucleotide comprises at least one modified sugar, modified nucleic acid base, or modified nucleoside bond with respect to the unmodified RNA or DNA. In certain embodiments, the oligonucleotide has a modified nucleoside. The modified nucleoside may comprise a modified sugar, a modified nucleic acid base, or both a modified sugar and a modified nucleic acid base. The modified oligonucleotide may also comprise terminal modifications, such as 5'-terminal modifications and 3'-terminal modifications.

[0227] Sugar modification and motifs In certain embodiments, the modified sugar is a substituted furanosyl sugar or a non-bicyclic modified sugar. In certain embodiments, the modified sugar is a bicyclic or tricyclic modified sugar. In certain embodiments, the modified sugar is an alternative sugar. The alternative sugar may contain one or more substitutions described herein.

[0228] In certain embodiments, the modified sugar is a substituted furanosyl or a non-bicyclic modified sugar. In certain embodiments, the furanosyl sugar is a ribosyl sugar. In certain embodiments, the furanosyl sugar contains one or more substituents, including but not limited to substituents at the 2', 3', 4', and 5' positions.

[0229] In certain embodiments, the substituent at the 2' position may include, but is not limited to, F and OCH3 ("OMe", "O-methyl", or "methoxy"). In certain embodiments, the substituent at the 2' position that is suitable for non-bicyclic modified sugars may include, but is not limited to, halo, allyl, amino, azide, SH, CN, OCN, CF3, OCF3, F, Cl, Br, SCH3, SOCH3, SO2CH3, ONE2, NO2, N3, and NH2. In certain embodiments, the substituent at the 2' position may include O-(C 1-C 10 Examples include, but are not limited to, alkoxy, alkoxyalkyl, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, O-alkynyl, S-alkynyl, N-alkynyl, O-alkyl-O-alkyl, and alkynyl, and the alkyl, alkenyl and alkynyl are substituted or unsubstituted C1-C 10 Alkyl, or C2-C 10 It can be an alkenyl or alkynyl. In certain embodiments, the substituent at the 2' position may be alkaryl, aralkyl, O-alkaryl and O-aralkyl, but is not limited to these. In certain embodiments, these 2' substituents may be further independently substituted with one or more substituents selected from hydroxyl, alkoxy, carboxy, benzyl, phenyl, nitro(NO2), thiol, thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl and alkynyl. In certain embodiments, the substituent at the 2' position may be O[(CH2) n O] m CH3, O(CH2) n OCH3, O(CH2) n CH3, O(CH2) n ONH2, O(CH2) n NH2, O(CH2) n SCH3 and O(CH2) n ON[(CH2) n Examples of substituents at the 2' position include, but are not limited to, OCH2CH2OCH3 ("MOE"), O(CH2)2ON(CH3)2 ("DMAOE"), O(CH2)2O(CH2)2N(CH3)2 ("DMAEOE"), and OCH2C(=O)-N(H)CH3 ("NMA"), but are not limited to these.

[0230] In certain embodiments, suitable substituents for the 4' position of a non-bicyclic modified sugar include, but are not limited to, alkoxy (e.g., methoxy), alkyl, and groups described in Manoharan et al. WO2015 / 106128. In certain embodiments, suitable substituents for the 5' position of a non-bicyclic modified sugar include, but are not limited to, methyl ("Me") (R or S), vinyl, and methoxy. In certain embodiments, one or more sugars include 5'-vinylphosphonate modification. In certain embodiments, the substituents described herein for the 2', 4', and 5' positions may be added to other specific positions of the sugar. In certain embodiments, such substituents may be added to the 3'-terminal nucleoside at the 3' position of the sugar, or to the 5'-terminal nucleoside at the 5' position. In certain embodiments, a non-bicyclic modified sugar may contain more than one non-crosslinked sugar substituent. In certain such embodiments, substituents on the non-bicyclic modified sugar include, but are not limited to, 5'-Me-2'-F and 5'-Me-2'-OMe (including both R and S isomers). In certain embodiments, substituents on the modified sugar include the groups described in WO2008 / 101157 by Migawa et al. and US2013 / 0203836 by Rajeev et al.

[0231] In certain embodiments, the modified sugar is a bicyclic sugar. A bicyclic sugar is a modified sugar containing two rings, the second ring forming a bicyclic structure by being formed via a bridge that links two atoms in the first ring. In certain embodiments, the bicyclic sugar contains a crosslinkable substituent that crosslinks two atoms of its furanosyl ring to form the second ring. In certain embodiments, the bicyclic sugar does not contain a furanosyl moiety. A "bicyclic nucleoside" ("BNA") is a nucleoside having a bicyclic sugar. In certain embodiments, the bicyclic sugar contains a bridge between the 4' and 2' furanose ring atoms. In certain embodiments, the bicyclic sugar contains a bridge between the 5' and 3' furanose ring atoms. In certain such embodiments, the furanose ring is a ribose ring. In certain embodiments, the 4' and 2' crosslinkable substituents include 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2' ("LNA"), 4'-CH2-S-2', 4'-(CH2)2-O-2' ("ENA"), 4'-CH(CH3)-O-2' ("restricted ethyl" or "cEt" when in S configuration), 4'-CH2-O-CH2-2', 4'-CH2-N(R)-2', 4'-CH(CH2OCH3)-O-2' ("restricted ethyl"). 4'-C(CH3)(CH3)-O-2' and its analogs (e.g., U.S. Patent No. 7,399,845), 4'-CH2-N(OCH3)-2' and its analogs (e.g., U.S. Patent No. 8,278,283), 4'-CH2-ON(CH3)-2' (e.g., U.S. Patent Application Publication No. 2004 / 0171570), 4'-CH2-N(R)-O-2' (wherein R is H, C1-C 12Examples include, but are not limited to, alkyl or protecting groups (e.g., U.S. Patent No. 7,427,672), 4'-CH2-C(H)(CH3)-2' (e.g., Chattopadhyaya el al., J. Org. Chem., 2009, 74, 118-134), and 4'-CH2-C(=CH2)-2' and its analogues (e.g., U.S. Patent No. 8,278,426). The entire contents of each of the above are incorporated herein by reference. Additional representative U.S. patents and U.S. patent application publications teaching the preparation of bicyclic nucleic acid nucleotides include U.S. Patents No. 6,268,490, 6,525,191, 6,670,461, 6,770,748, 6,794,499, 6,998,484, 7,053,207, 7,034,133, 7,084,125, 7,399,845, 7,427,672, and 7 Examples include, but are not limited to, US2008 / 0039618, US2009 / 0012281, US2013 / 0190383, and WO2013 / 036868, the entire contents of which are incorporated herein by reference. Any of the above bicyclic nucleosides having one or more stereochemical sugar configurations (e.g., including α-L-ribofuranose and β-D-ribofuranose) can be prepared (see, for example, WO99 / 14226). Unless otherwise specified, the specific bicyclic nucleoside in this invention has a β-D configuration.

[0232] In certain embodiments, the modified sugar is an alternative sugar. In certain embodiments, the alternative sugar has its oxygen atom replaced by, for example, a sulfur atom, a carbon atom, or a nitrogen atom. In certain such embodiments, the alternative sugar may also include crosslinkable substituents and / or non-crosslinkable substituents as described herein. In certain embodiments, the alternative sugar includes a ring having more than five atoms. In certain such embodiments, the alternative sugar includes a cyclobutyl moiety instead of pentofuranosyl sugar. In certain embodiments, the alternative sugar includes a six-membered ring instead of pentofuranosyl sugar. In certain embodiments, the alternative sugar includes tetrahydropyran ("THP") instead of pentofuranosyl sugar. In certain embodiments, the alternative sugar includes morpholino instead of pentofuranosyl sugar. Representative U.S. patents teaching the preparation of such modified sugar structures include U.S. Patents No. 4,981,957, No. 5,118,800, No. 5,166,315, No. 5,185,444, No. 5,319,080, No. 5,359,044, No. 5,393,878, No. 5,446,137, No. 5,466,786, No. 5,514,785, No. 5,519,134, No. 5,567,811, No. 5,576,427, No. 5,591,722, and Nos. 5,597,909, 5,610,300, 5,627,053, 5,639,873, 5,646,265, 5,658,873, 5,670,633, 5,700,920, 7,875,733, 7,939,677, 8,088,904, 8,440,803, and 9,005,906 are examples of, but are not limited to, the overall contents of each of the above are incorporated herein by reference.

[0233] In some embodiments, the alternative sugar includes an acyclic moiety. In certain embodiments, the alternative sugar is an unlocked nucleic acid ("UNA"). A UNA is an acyclic nucleic acid of unlocked, in which one of the bonds of its sugar is removed to form the "sugar" residue of unlocked. In one example, a UNA also includes a monomer in which the bond between C1' and C4' (i.e., the carbon-oxygen-carbon covalent bond between the C1' and C4' carbons) is removed. In another example, the bond between C2' and C3' of the sugar (i.e., the carbon-carbon covalent bond between the C2' and C3' carbons) is removed. Representative U.S. publications teaching the preparation of UNAs include, but are not limited to, U.S. Patent No. 8,314,227, and U.S. Patent Application Publications 2013 / 0096289, 2013 / 0011922, and 2011 / 0313020, the entire contents of each of these are incorporated herein by reference. In certain embodiments, alternative sugars include peptide nucleic acids ("PNAs"), acyclic butyl nucleic acids (see, e.g., Kumar et al., Org. Biomol. Chem., 2013, 11, 5853-5865), and nucleosides and oligonucleotides described in Manoharan et al., U.S. 2013 / 130378, the entire contents of these are incorporated herein by reference. Many other bicyclic and tricyclic sugars, as well as cyclic systems of alternative sugars, that can be used with modified nucleosides are known in the art.

[0234] In certain embodiments, the disclosure relates to a compound comprising at least one oligonucleotide, the nucleoside of such oligonucleotide comprising one or more types of modified sugars and / or unmodified sugars arranged along the oligonucleotide or region thereof in a defined pattern, i.e., a “sugar motif”. In certain cases, such sugar motifs include, but are not limited to, any of the sugar modification patterns described herein.

[0235] In certain embodiments, the oligonucleotide contains a gapmer sugar motif. The gapmer oligonucleotide contains, or comprises, a region having two outer "wing" regions and a central or inner "gap" region. The gap and wing regions form a continuous sequence of nucleosides, where the majority of the nucleoside sugars in each wing differ from the majority of the nucleoside sugars in the gap. In certain embodiments, the wing regions contain mostly modified sugars, and the gap contains mostly unmodified sugars. In certain embodiments, the nucleosides in the gap are deoxynucleosides. Compounds having a gapmer sugar motif are described, for example, in U.S. Patent No. 8,790,919, the contents of which are incorporated herein by reference.

[0236] In certain embodiments, one or both oligonucleotides of a double-stranded compound contain a triple sugar motif. An oligonucleotide having a triple sugar motif contains three identical sugar modifications on three consecutive nucleosides. In certain embodiments, the triple sugar is located at or near the cleavage site of the oligonucleotide. In certain embodiments, an oligonucleotide of a double-stranded compound may contain more than one triple sugar motif. In certain embodiments, the identical sugar modifications of the triple sugar motif are 2'-F modifications. Compounds having triple sugar motifs are disclosed, for example, in U.S. Patent No. 10,668,170, the contents of which are incorporated herein by reference.

[0237] In certain embodiments, one or both oligonucleotides of a double-stranded compound contain a quadruple sugar motif. An oligonucleotide having a quadruple sugar motif contains four identical sugar modifications on four consecutive nucleosides. In certain embodiments, the quadruple sugar is located at or near its cleavage site. In certain embodiments, an oligonucleotide of a double-stranded compound may contain more than one quadruple sugar motif. In certain embodiments, the identical sugar modifications on the quadruple sugar motif are 2'-F modifications. In double-stranded compounds having a double-stranded region with nucleotide lengths of 19–23, the cleavage sites of the antisense oligonucleotide are typically located around positions 10, 11, and 12 from its 5' end. In certain embodiments, the quadruple sugar motif is located at positions 8, 9, 10, 11; 9, 10, 11, 12; 10, 11, 12; 10, 11, 12, 13; 11, 12, 13, 14, or 12, 13, 14, 15 of the sense oligonucleotide, counting from the first nucleoside at the 5' end of the sense oligonucleotide, or counting from the first paired nucleotide in the double-stranded region at the 5' end of the sense oligonucleotide. In certain embodiments, the quadruple sugar motif is located at positions 8, 9, 10, 11; 9, 10, 11, 12; 10, 11, 12; 10, 11, 12, 13; 11, 12, 13, 14, or 12, 13, 14, 15 of the antisense oligonucleotide, counting from the first nucleoside at the 5' end of the antisense oligonucleotide, or counting from the first paired nucleotide in the double-stranded region from the 5' end of the antisense oligonucleotide. The cleavage sites may vary depending on the length of the double-stranded region of the double-stranded compound, and the position of the quadruple sugar may change accordingly.

[0238] In certain embodiments, the oligonucleotide contains alternating sugar motifs. In certain embodiments, one or both oligonucleotides of a double-chain compound contain alternating sugar motifs. The oligonucleotide having alternating sugar motifs contains at least two different sugar modifications, where one or more consecutive nucleosides containing a first sugar modification are alternated with, for example, one or more consecutive nucleosides containing a second sugar modification and one or more consecutive nucleosides containing a third sugar modification, and so on. For example, if A, B, and C each represent one type of modification to the nucleoside, the alternating motifs could be "ABABABABABAB...", "AABBAABBAABB...", "AABAABAABAAB", "AAABAAABAAAB...", "AAABBBAAABBB...", or "ABCABCABCABC...". In certain embodiments, the alternating sugar motif is repeated along an oligonucleotide over at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23 consecutive nucleic acid bases. In certain embodiments, the alternating sugar motif is composed of two different sugar modifications. In certain embodiments, the alternating sugar motif includes 2'-OMe and 2'-F sugar modifications.

[0239] In certain embodiments, each nucleoside of an oligonucleotide is independently modified with one or more sugar modifications provided in this disclosure. In certain embodiments, each oligonucleotide of a double-stranded compound independently has one or more sugar motifs provided in this disclosure. In certain embodiments, an oligonucleotide containing a sugar motif is fully modified in such a way that each nucleoside other than the nucleoside containing the sugar motif contains a sugar modification.

[0240] Nucleic acid base modification and motifs In certain embodiments, the modified oligonucleotide comprises one or more nucleosides containing modified nucleic acid bases. In certain embodiments, the modified oligonucleotide comprises one or more nucleosides that do not contain nucleic acid bases (referred to as debasalized nucleosides).

[0241] In certain embodiments, the modified nucleic acid bases are selected from 5-substituted pyrimidines, 6-azapyrimidines, alkyl or alkynyl-substituted pyrimidines, alkyl-substituted purines, and N-2, N-6, and O-6-substituted purines. In certain embodiments, the modified nucleic acid bases are 2-aminopropyladenine, 5-hydroxymethylcytosine, 5-methylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine, 2-propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl(C≡C-CH3)uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azocymine, 5-ribosyluracil (pseudracil), 4-thiouracil, 8-haloprin, 8-aminopurine, 8-thiolpurine, 8-thioalkylpurine, 8-hydroxylpurine, and 8-azocytosine. The bases are selected from purines and other 8-substituted purines, 5-halo (especially 5-bromo), 5-trifluoromethyl, 5-halouracil and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl4-N-benzoylcytosine, 5-methyl4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous bases, size-extended bases, and fluorinated bases. Further modified nucleic acid bases include tricyclic pyrimidines, such as 1,3-diazaphenoxadin-2-one, 1,3-diazaphenothiazine-2-one, and 9-(2-aminoethoxy)-1,3-diazaphenoxadin-2-one (G-clamp). Other modified nucleic acid bases may include bases in which the purine or pyrimidine base is replaced by another heterocycle, such as 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine, and 2-pyridone.

[0242] Further nucleic acid bases are listed in U.S. Patent No. 3,687,808, Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed. Wiley-VCH, 2008, The Concise Encyclopedia Of Polymer Science And Engineering, pages 858-859, Kroschwitz, JL, Ed., John Wiley & Sons, 1990, 858-859, Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613, Sanghvi, YS, Chapter 15, dsRNA Research and Applications, pages 289-302, Antisense Research and Applications, Crooke, ST and Lebleu, B., Eds., CRC Press, 1993, 273-288, Antisense Drug Technology, Crooke ST, Ed., CRC The bases disclosed in Press, 2008, pp. 163-166 and pp. 442-443 (Chapters 6 and 15) are listed, each incorporated herein by reference.

[0243] Publications teaching the preparation of specific modified nucleic acid bases shown above, as well as other modified nucleic acid bases, include U.S. Patent Applications Publications 2003 / 0158403 and 2003 / 0175906, U.S. Patents 4,845,205, 5,130,302, 5,134,066, 5,175,273, 5,367,066, and 5,432,272. , No. 5,434,257, No. 5,457,187, No. 5,459,255, No. 5,484,908, No. 5,502,177, No. 5,525,711, No. 5,5 No. 52,540, No. 5,587,469, No. 5,594,121, No. 5,596,091, No. 5,614,617, No. 5,645,985, No. 5,681,941 No. 5,811,534, No. 5,750,692, No. 5,948,903, No. 5,587,470, No. 5,457,191, No. 5,763,588, No. 5, No. 830,653, No. 5,808,027, No. 6,005,096, No. 6,015,886, No. 6,147,200, No. 6,166,197, No. 6,166,19 Nos. 9, 6,222,025, 6,235,887, 6,380,368, 6,528,640, 6,639,062, 6,617,438, 7,045,610, 7,427,672, and 7,495,088 are examples of, but are not limited to, the contents of which are incorporated herein by reference.

[0244] In certain embodiments, the oligonucleotide comprises modified and / or unmodified nucleic acid bases arranged in a defined pattern or motif along the oligonucleotide or its region. In certain embodiments, each nucleic acid base is modified. In certain embodiments, none of the nucleic acid bases are modified. In certain embodiments, each purine or pyrimidine is modified. In certain embodiments, each adenine is modified. In certain embodiments, each guanine is modified. In certain embodiments, each thymine is modified. In certain embodiments, each uracil is modified. In certain embodiments, each cytosine is modified. In certain embodiments, some or all of the cytosine nucleic acid bases in the modified oligonucleotide are 5-methylcytosine.

[0245] In certain embodiments, the modified oligonucleotide includes a block of modified nucleic acid bases. In certain such embodiments, the block is located at the 3' end of the oligonucleotide. In certain embodiments, the block is within 3 nucleosides of the 3' end of the oligonucleotide. In certain embodiments, the block is located at the 5' end of the oligonucleotide. In certain embodiments, the block is within 3 nucleosides of the 5' end of the oligonucleotide.

[0246] Modification and motifs of internucleoside bonds The 3'→5' phosphodiester bond is a natural nucleoside bond in RNA and DNA. In certain embodiments, oligonucleotides have one or more modified nucleotide bonds, i.e., unnatural nucleotide bonds. Certain unnatural nucleoside bonds may confer desirable properties, such as enhanced cellular uptake, increased affinity for target nucleic acids, and improved stability in the presence of nucleases. Representative phosphorus-containing modified nucleoside bonds include, but are not limited to, phosphotriesters, alkylphosphonates (e.g., methylphosphonates), phosphoramidates, and phosphorothioates ("P=S") and phosphorodithioates ("HS-P=S"). Representative non-phosphorus-containing internucleoside bonds include, but are not limited to, methylenemethylimino (-CH2-N(CH3)-O-CH2), thiodiesters, thionocarbamates (-OC(=O)(NH)-S-), siloxanes (-O-SiH2-O-), and N,N'-dimethylhydrazine (-CH2-N((CH3)-N((CH3)-). Methods for preparing phosphorus-containing and non-phosphorus-containing internucleoside bonds are well known to those skilled in the art. Examples of neutral internucleoside bonds include phosphotriesters, methylphosphonates, and MMI(3'-CH2-N(C Examples of neutral internucleoside bonds include, but are not limited to, H3)-O-5'), amide-3(3'-CH2-C(=O)-N(H)-5'), amide-4(3'-CH2-N(H)-C(=O)-5'), formacetal(3'-O-CH2-O-5'), methoxypropyl, and thioformacetal(3'-S-CH2-O-5'). Further examples of neutral internucleoside bonds include nonionic bonds, such as siloxanes (dialkylsiloxanes), carboxylic acid esters, carboxamides, sulfides, sulfonic acid esters, and amides (see, for example, Carbohydrate Modifications in Antisense Research; YSSanghvi and PDCook, Eds., ACS Symposium Series 580; Chapters 3 and 4, 40-65).Further examples of neutral nucleoside bonds include nonionic bonds containing mixed N, O, S, and CH2 component parts.

[0247] In certain embodiments, the oligonucleotide includes at least one modified nucleoside bond. The modified nucleoside bond may be located at any of the positions of the oligonucleotide. In double-stranded compounds, the modified nucleoside bond may be located within the sense oligonucleotide, the antisense oligonucleotide, or both of the oligonucleotides of the double-stranded compound.

[0248] In certain embodiments, the nucleoside-to-nucleoside modification may occur on all nucleosides of the oligonucleotide. In certain embodiments, the nucleoside-to-nucleoside modification may occur in an alternating pattern along the oligonucleotide. In certain embodiments, each nucleoside-to-nucleoside bond is essentially a phosphate nucleoside bond (P=O). In certain embodiments, each nucleoside-to-nucleoside bond in the modified oligonucleotide is a phosphorothioate (P=S). In certain embodiments, each nucleoside-to-nucleoside bond in the modified oligonucleotide is independently selected from phosphorothioate and phosphate nucleoside bonds. In certain embodiments, the pattern of nucleoside-to-nucleoside modification is the same in each oligonucleotide of the double-stranded compound. In certain embodiments, the pattern of nucleoside-to-nucleoside modification is different in each oligonucleotide of the double-stranded compound. In certain embodiments, the double-stranded compound contains 6 to 8 modified nucleoside bonds. In certain embodiments, the 6 to 8 modified nucleoside bonds are phosphorothioate nucleoside bonds or alkylphosphonate nucleoside bonds. In certain embodiments, the sense oligonucleotide contains at least two modified nucleoside bonds at either or both of its 5' and 3' ends. In certain such embodiments, the modified nucleoside bonds are phosphorothioate nucleoside bonds or alkylphosphonate nucleoside bonds. In certain embodiments, the antisense oligonucleotide contains at least two modified nucleoside bonds at either or both of its 5' and 3' ends. In certain such embodiments, the modified nucleoside bonds are phosphorothioate nucleoside bonds or alkylphosphonate nucleoside bonds.

[0249] In certain embodiments, the double-stranded compound includes an overhang region. In certain embodiments, the double-stranded compound includes a phosphorothioate or alkylphosphonate nucleoside linkage modification in its overhang region. In certain embodiments, the double-stranded compound includes a phosphorothioate nucleotide linkage or alkylphosphonate nucleotide linkage that links its overhang nucleotide to the adjacent paired nucleotide. For example, there may be at least two phosphorothioate nucleoside links between three terminal nucleosides, two of which are overhang nucleosides, and the third is the adjacent paired nucleoside. These three terminal nucleosides may be at the 3' end of the antisense oligonucleotide, the 3' end of the sense oligonucleotide, the 5' end of the antisense oligonucleotide, or the 5' end of the antisense oligonucleotide.

[0250] In certain embodiments, the modified oligonucleotide contains one or more nucleoside bonds having a chiral center. Typical chiral nucleoside bonds include, but are not limited to, alkylphosphonates and phosphorothioates. Modified oligonucleotides containing nucleoside bonds having a chiral center can be prepared in a specific stereochemical configuration as a group of modified oligonucleotides containing stereorandom nucleoside bonds or as a group of modified oligonucleotides containing phosphorothioate bonds. In certain embodiments, the group of modified oligonucleotides contains phosphorothioate nucleoside bonds, all of which are stereorandom. Such modified oligonucleotides can be produced using a synthetic method that randomly selects the stereochemical configuration of each phosphorothioate bond. As will be well understood by those skilled in the art, each individual phosphorothioate of each individual oligonucleotide molecule has a defined stereochemical configuration. In certain embodiments, the population of modified oligonucleotides is enriched for modified oligonucleotides containing one or more specific phosphorothioate nucleoside bonds in a specific, independently selected stereochemical configuration. In certain embodiments, the specific configuration of the particular phosphorothioate bond is present in at least 65% of the molecules in the population. In certain embodiments, the specific configuration of the particular phosphorothioate bond is present in at least 70% of the molecules in the population. In certain embodiments, the specific configuration of the particular phosphorothioate bond is present in at least 80% of the molecules in the population. In certain embodiments, the specific configuration of the particular phosphorothioate bond is present in at least 90% of the molecules in the population. In certain embodiments, the specific configuration of the particular phosphorothioate bond is present in at least 99% of the molecules in the population. Such enriched populations of modified oligonucleotides can be produced using synthetic methods known in the art, for example, the methods described in Oka et al., JACS 125, 8307 (2003), Wan et al. Nuc. Acid. Res. 42, 13456 (2014), and WO2017 / 015555.In certain embodiments, the population of modified oligonucleotides is enriched with respect to modified oligonucleotides having at least one phosphorothioate in the (Sp) conformation. In certain embodiments, the population of modified oligonucleotides is enriched with respect to modified oligonucleotides having at least one phosphorothioate in the (Rp) conformation.

[0251] NMDA receptor ligand In some embodiments, the compounds provided in the present invention include an NMDA receptor ligand. In some embodiments, the NMDA receptor ligand is useful for inducing a therapeutic, prophylactic, or diagnostic agent. In certain embodiments, the therapeutic agent is an oligonucleotide (e.g., a therapeutic oligonucleotide). In some embodiments, the NMDA receptor ligand induces the oligonucleotide locally. In some embodiments, the NMDA receptor ligand targets a tissue. In some embodiments, that tissue is brain tissue. In some embodiments, the NMDA receptor ligand targets a cell receptor. In some embodiments, the cell receptor is an NMDA receptor. In some embodiments, the NMDA receptor is located in the brain. In some embodiments, the NMDA receptor is located in the frontal cortex. In some embodiments, the NMDA receptor is located in the striatum. In some embodiments, the NMDA receptor is located in the cerebellum. In some embodiments, the NMDA receptor is located in the brainstem. In some embodiments, the NMDA receptor is located in the hippocampus. In some embodiments, the NMDA receptor is located in the spinal cord.

[0252] This disclosure assumes the use of any of the NMDA receptor ligands in the compounds provided in the present invention. NMDA receptor ligands are known in the art, and those skilled in the art will be able to identify additional NMDA receptor ligands for use in the compounds described herein, other than those explicitly shown in this disclosure. This disclosure also assumes the use of derivatives and prodrugs of any NMDA receptor ligand provided in the present invention or known in the art in the compounds described herein, and those skilled in the art will know how to prepare such derivatives and prodrugs.

[0253] In some embodiments, the NMDA receptor ligand is an NMDA receptor agonist. In some embodiments, the NMDA receptor ligand is an NMDA receptor antagonist. In some embodiments, the NMDA receptor ligand is memantine, MK-801, huperzine (e.g., huperzine A, huperzine B), cholesterol, lacosamide, lapastinel, ketamine, or a structural analog or derivative thereof. In some embodiments, the NMDA receptor ligand is one of those described in Neuropharmacology 2007, 53(6), 699-723, J.Med.Chem. 1990, 33(2), 789-808, Neuroscience 2001, 105(3), 663-669, J.Med.Chem. 2022, 65(13), 9063-9075, Drugs Fut. 2004, 29(10), 992, Drugs Fut. 2004, 29(10), 993, and British Journal of Pharmacology 2022, 179(6), 1146-1187, each of which is incorporated herein by reference. Examples of NMDA receptor ligands for use in this disclosure include: [ka] Examples include, but are not limited to, any of the NMDA receptor ligands and their derivatives.

[0254] In some embodiments, the NMDA receptor ligand is an anti-NMDA receptor antibody. In certain embodiments, the NMDA receptor ligand is an anti-NMDA receptor antibody fragment or an anti-NMDA receptor antibody variant. "Anti-NMDA receptor antibody" refers to an immune system protein that recognizes the NMDA receptor, binds to the NMDA receptor, or interacts with the NMDA receptor in a different way.

[0255] In certain embodiments, the NMDA receptor ligand is conjugated (e.g., linked, connected, bound, or associated) with one or more drug moieties. In certain embodiments, the drug moieties are therapeutic agents, prophylactic agents, diagnostic agents, or imaging agents. In certain embodiments, the drug is a small molecule or an oligomeric compound. In certain embodiments, the drug moieties are proteins, peptides, antibodies, oligonucleotides, small molecules, macromolecules, or combinations thereof.

[0256] In some embodiments, more than one NMDA receptor ligand is conjugated to the drug moiety. In some embodiments, at least two NMDA receptor ligands (e.g., two, three, four, five, six, seven, eight, nine, or ten or more NMDA receptor ligands) are conjugated to the drug moiety. In some embodiments, two NMDA receptor ligands are conjugated to the drug moiety. In some embodiments, three NMDA receptor ligands are conjugated to the drug moiety. In some embodiments, four NMDA receptor ligands are conjugated to the drug moiety. In some embodiments, five NMDA receptor ligands are conjugated to the drug moiety. In some embodiments, more than five NMDA receptor ligands are conjugated to the drug moiety. In some embodiments, at least one to about five NMDA receptor ligands are conjugated to the drug moiety. In some embodiments, at least one to about four NMDA receptor ligands are conjugated to the drug moiety. In some embodiments, at least one to about three NMDA receptor ligands are conjugated to the drug moiety. In some embodiments, at least one to about two NMDA receptor ligands are conjugated to the drug moiety.

[0257] When the drug moiety is conjugated to multiple NMDA receptor ligands, all of the NMDA receptor ligands may be conjugated at the same position or near the same position on the drug moiety, or they may be conjugated at multiple different positions on the drug moiety.

[0258] In some embodiments, an oligonucleotide is conjugated (e.g., linked, bound, associated) to an NMDA receptor ligand either through either the 5' end and / or the 3' end of the oligonucleotide, or at an internal position within the oligonucleotide (i.e., a nucleotide of the oligonucleotide other than the 5' or 3' nucleotide). In some embodiments, the oligonucleotide is conjugated to an NMDA receptor ligand through the 5' end of the oligonucleotide. In some embodiments, the oligonucleotide is conjugated to an NMDA receptor ligand through the 3' end of the oligonucleotide. In some embodiments, the oligonucleotide is conjugated to an NMDA receptor ligand through both the 5' and 3' ends of the oligonucleotide. In some embodiments, the oligonucleotide is conjugated to an NMDA receptor ligand at an internal position within the oligonucleotide (e.g., at an "internally modified oligonucleotide").

[0259] In some embodiments, the oligonucleotide is conjugated to more than one NMDA receptor ligand. In some embodiments, the oligonucleotide is conjugated to at least two NMDA receptor ligands (e.g., two, three, four, five, six, seven, eight, nine, or ten or more NMDA receptor ligands). In some embodiments, the oligonucleotide is conjugated to two NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated to three NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated to four NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated to five NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated to more than five NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated to at least one to about five NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated to at least one to about four NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated to at least one to about three NMDA receptor ligands. In some embodiments, the oligonucleotide is conjugated to at least one to about two NMDA receptor ligands.

[0260] When an oligonucleotide is conjugated with multiple NMDA receptor ligands, all of the NMDA receptor ligands may be conjugated at or near the same position on the oligonucleotide, or the NMDA receptor ligands may be conjugated at multiple different positions on the oligonucleotide. In some embodiments, multiple NMDA receptor ligands (i.e., two, three, four, or five or more NMDA receptor ligands) are conjugated at the 5' end of the oligonucleotide. In some embodiments, multiple NMDA receptor ligands (i.e., two, three, four, or five or more NMDA receptor ligands) are conjugated at the 3' end of the oligonucleotide. In some embodiments, multiple NMDA receptor ligands (i.e., two, three, four, or five or more NMDA receptor ligands) are conjugated at one or more internal positions on the oligonucleotide. In some embodiments, an oligonucleotide is conjugated to one or more NMDA receptor ligands at its 5' end, and / or to one or more NMDA receptor ligands at its 3' end, and / or to one or more NMDA receptor ligands at an internal or multiple internal position of the oligonucleotide.

[0261] Linker In certain embodiments, a conjugate of a compound formula described herein is provided. In certain embodiments, the conjugate comprises an NMDA receptor ligand covalently bonded to a drug portion. In certain embodiments, the conjugate provided herein comprises one or more linker portions. In certain embodiments, the one or more linker portions link the NMDA receptor ligand to the drug portion. In certain embodiments, the drug portion is a protein, peptide, antibody, nucleic acid, small molecule, macromolecule, therapeutic agent, prophylactic agent, diagnostic agent, or imaging agent. In some embodiments, the compound is conjugated to an oligonucleotide. In certain embodiments, the NMDA receptor ligand is conjugated to an oligonucleotide. In certain embodiments, the compound comprises one or more NMDA receptor ligands, one or more linker portions, and one or more drug portions, wherein the NMDA receptor ligand is conjugated (e.g., linked, connected, bound, associated) to one or more drug portions through one or more linker portions.

[0262] Conjugates such as those disclosed herein can be manufactured using any available method. When associating the compound provided in the present invention with a drug moiety (e.g., an NMDA receptor ligand with an oligonucleotide), the moiety is defined as a linker moiety, i.e., the linker is covalently bonded to the oligonucleotide and the NMDA receptor ligand, respectively, and in some of the formulas of the present invention, it is defined as "-L nThe formula is "-", and the components may be linked directly or indirectly (through a portion where n is a number (e.g., L1, L2, L3, L4)). For example, the oligonucleotide and the NMDA receptor ligand may be directly linked to each other, for example, by one or more covalent bonds, or by one or more linkers. "Linker" refers to any chemical portion (e.g., a combination of atoms whose valence is appropriate according to known chemical principles) used for the purpose of conjugating two components of the compound provided in the present invention (e.g., the NMDA receptor ligand and the oligonucleotide) to each other. Each of the two components may be connected to any portion of any linker provided in the present invention. In some embodiments, one of the components of the compound provided in the present invention (e.g., the NMDA receptor ligand or the oligonucleotide) is connected by a bond to one end of a linker, and the other component is connected by a bond to the other end of that linker. In some embodiments, one or both components of the compound provided in the present invention may be connected by a bond to an internal position of any of the linkers described herein. For example, with respect to an alkyl linker, the NMDA receptor ligand may be conjugated to the carbon at one end of the alkyl linker, and the oligonucleotide may be conjugated to the carbon at the other end of the alkyl linker. In some embodiments, the linker is a bond (e.g., a phosphodiester bond and a phosphorothioate bond). In some embodiments, the linker is an optionally substituted alkyl linker (i.e., two parts are linked using an alkyl chain, where each of these parts may be conjugated to the opposite end of the alkyl linker, or one or both parts may be conjugated to a carbon inside the alkyl linker).In some embodiments, the linker is an optionally substituted polyethylene glycol (PEG) linker (i.e., two parts are joined using a PEG chain, each of which may be conjugated to an opposing end of the PEG linker, or one or both parts may be conjugated to an internal position on the PEG linker). In some embodiments, the linker is an optionally substituted heteroalkyl linker (i.e., two parts are joined using a heteroalkyl chain, each of which may be conjugated to an opposing end of the heteroalkyl linker, or one or both parts may be conjugated to an internal position on the heteroalkyl linker). In some embodiments, the linker is an optionally substituted heteroaryl linker (i.e., two parts are joined using a heteroaryl group, each of which may be conjugated to any of the positions of the heteroaryl group).

[0263] In certain embodiments, the compounds provided in the present invention include one or more linking groups. In certain embodiments, L1, L2, L3, and L4 each include a linking group. In certain embodiments, the linking group is covalently bonded to an NMDA receptor ligand. In certain embodiments, the linking group is covalently bonded to an oligonucleotide. In certain embodiments, the linking group is covalently bonded to a cleavable portion. In certain embodiments, the linking group includes a cleavable bond. In certain embodiments, the linking group does not include a cleavable portion. In certain embodiments, the linking group includes a covalent bond to a solid support. In certain embodiments, the linking group includes multiple positions for binding to an NMDA receptor ligand.

[0264] In certain embodiments, the linking group includes repeating units of a chain structure, such as a hydrocarbyl chain, or an oligomer, or a combination of such repeating units. In certain embodiments, the linking group includes 1 to 50 repeating units, 1 to 40 repeating units, 1 to 25 repeating units, 1 to 20 repeating units, 1 to 15 repeating units, 1 to 10 repeating units, or 1 to 5 repeating units. In certain embodiments, the linking group has an atomic length of 1 to 50, an atomic length of 1 to 40, an atomic length of 1 to 25, an atomic length of 1 to 20, an atomic length of 1 to 15, an atomic length of 1 to 10, or an atomic length of 1 to 5.

[0265] In certain embodiments, the linking group includes a carbon atom. In certain embodiments, the linking group includes a heteroatom (e.g., nitrogen, oxygen, sulfur, etc.). In certain embodiments, the linking group forms an amide bond, an ester bond, or a disulfide bond. In certain embodiments, the linking group forms a hydrazone bond, an oxime bond, an imine bond, a guanidine bond, a urea bond, a carbamate bond, an unsaturated alkyl bond, a sulfonamide bond, or a 4-8 membered heterocyclic bond. In certain embodiments, the linking group includes one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In certain embodiments, the linking group includes at least one phosphorus group. In certain embodiments, the linking group includes at least one phosphate group. In certain embodiments, the linking group includes at least one neutral linking group. In certain embodiments, the linking group is substituted with various substituents, including, but not limited to, hydrogen atom groups, alkyl groups, alkenyl groups, alkynyl groups, amino groups, alkylamino groups, dialkylamino groups, trialkylamino groups, hydroxyl groups, alkoxy groups, halogen groups, aryl groups, heterocyclic groups, aromatic heterocyclic groups, cyano groups, amide groups, carbamoyl groups, carboxylic acid groups, ester groups, thioether groups, alkylthioether groups, thiol groups, and ureido groups. As will be apparent to those skilled in the art, each of these groups may be further substituted.

[0266] In certain embodiments, the linking group may be a substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C2-C 10 Alkenyl, or substituted or unsubstituted C2-C 10 Examples of preferred substituents include, but are not limited to, alkynyl substituents, and include hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl, and alkynyl substituents. In certain embodiments, the linking group is aliphatic or heteroaliphatic. For example, the linking group may be a polyalkyl linking group. The linking group may be a polyether linking group. The linking group may be a polyethylene linking group, such as PEG.

[0267] In certain embodiments, the linking group is a short peptide chain. In certain embodiments, the linking group contains 1 to 40 amino acids, 1 to 25 amino acids, 1 to 20 amino acids, 1 to 15 amino acids, 1 to 10 amino acids, or 1 to 5 amino acids.

[0268] In certain embodiments, the linking group comprises linker nucleosides. In certain embodiments, the linking group comprises 1 to 40 linker nucleosides, 1 to 25 linker nucleosides, 1 to 20 linker nucleosides, 1 to 15 linker nucleosides, 1 to 10 linker nucleosides, or 1 to 5 linker nucleosides. In certain embodiments, such linker nucleosides may be modified or unmodified nucleosides. It is typically desirable that the linker nucleosides be cleaved from the compound once the compound reaches the target tissue. Thus, the linker nucleosides in the present invention can be linked to each other and to the remainder of the compound via cleavable bonds. In the present invention, the linker nucleosides are not considered to be part of their oligonucleotide payload. Therefore, in embodiments in which the compound comprises an oligonucleotide consisting of a predetermined number or range of linked nucleosides and / or an oligonucleotide having a predetermined complementarity percentage to a reference nucleic acid, and the compound also comprises an NMDA receptor ligand having a linking group containing a linker-nucleoside, these linker-nucleosides are not counted in relation to the length of the oligonucleotide and are not used when determining the complementarity percentage of the oligonucleotide to the reference nucleic acid.

[0269] In certain embodiments, the linking group includes a protein-binding group. In certain embodiments, the protein-binding group is a lipid, for example, cholesterol, cholic acid, adamantane acetate, 1-pyrene butyric acid, dihydrotestosterone, 1,3-bis-O(hexadecyl)glycerol, geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, dimethoxytrityl or Examples include, but are not limited to, phenoxazine, vitamins (e.g., folic acid, vitamin A, vitamin E, biotin, pyridoxal), peptides, glycans (e.g., monosaccharides, disaccharides, trisaccharides, tetrasaccharides, oligosaccharides, polysaccharides), endosomal soluble components, steroids (e.g., ubaol, hecogenin, diosgenin), terpenes (e.g., triterpenes, e.g., sarsasapogenin, friederin, litocholic acid derivatized by epifriederanol), or cationic lipids. In certain embodiments, the protein-binding group has a chain length of C 16 ~C 22 These include saturated or unsaturated fatty acids, cholesterol, cholic acid, vitamin E, adamantane, or 1-pentafluoropropyl.

[0270] In certain embodiments, the linking group may include, but is not limited to, pyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), and 6-aminohexanoic acid (HEC or AHA).

[0271] The total amount of the product is US5,994,517, US6,300,319, US6,6 60,720, US6,906,182, US7,262,177, US7,491,805, US8.1 06,022, US7,723,509, US9,127,276, US2006 / 0148740, US2011 / 0123520, WO2013 / 033230, WO2012 / 037254, Biessen et al.,J.Med.Chem.1995,38,1846-1852;Lee et al.,Bioorganic & Medicinal Chemistry 2011,19,2494-2500;Rensen et al.,J.Biol.Chem.2001,276,37577-37584; al.,J.Med.Chem.2004,47,5798-5808;Sliedregt et al.,J.Med.Chem.1999,42,609-618; al.,Tetrahedron,1997,53,759-770;Lee,Carbohydr.Res.1978,67,509-514;Connolly et al.,J.Biol.Chem.1982,257,939-945;Pavia et al.,Int.J.Pep.Protein Res.1983,22,539-548;Lee et al.,Biochem.1984,23,4255-4261;Lee et al.,Glycoconjugate J.1987,4,317-328;Toyokuni et al.,Tetrahedron Lett.1990,31,2673-2676、Biessen et al.,J.Med.Chem.1995,38,1538-1546; Lett.1997,38,3487-3490;Lee et al.,Bioconjug.Chem.1997,8,762-765;Kato et al.,Glycobiol.2001,11,821-829;Rensen et al.,J.Biol.Chem.2001,276,37577-37584、Lee et al.,Methods Enzymol.2003,362,38-43、Westerlind et al.,Glycoconj.J.2004,21,227-241、Lee et al. al.,Bioorg.Med.Chem.Lett.2006,16(19),5132-5135、Maierhofer et al.,Bioorg.Med.Chem.2007,15,7661-7676、Khorev et al.,Bioorg.Med.Chem.2008,16,5216-5231、Lee et al.,Bioorg.Med.Chem.2011,19,2494-2500、Kornilova et al.,Analyt.Biochem.2012,425、F al.,Angew.Chemie Int.Ed.Engl.2012,51,7445-7448、Biessen et al.,J.Med.Chem.1995,38,1846-1852、Sliedregt et al.,J.Med.90-4,699 et2,699、 al.,J.Med.Chem.2004,47,5798-5808、Rensen et al.,Arterioscler.Thromh.Vase.Biol.2006,26,169-175、van Rossenberg et al.,Gene Ther.2004,11,457、 al.,J.Am.Chem.Soc.2004,126,14013-14022、Lee et al.,J.Org.Chem.2012,77,7564-7571、Biessen et al.,FASEB J.2000 et14,1722 al.,Bioconjug.Chem.1997,8,935-940、Duff et al.,Methods Enzymol.2000,313,297-321、Maier et al.,Bioconjug.Chem.2003,14,18-29、Jayaprakash et al. al.,Org.Lett.2010,12,5410-5413、Manoharan,Antisense Nucleic Acid Drug Dev.2002,12,103-128,Merwin et al.,Bioconjug.Chem.1994,5,612-620,Tomiya et al. al., Bioorg.Med.Chem., 2013, 21, 5275-5281, International Publication No. WO1998 / 013381, Same No. WO2011 / 038356, Same No. WO1997 / 046098, Same No. WO2008 / 098788, Same No. WO20 No. 04 / 101619, same as WO2012 / 037254, same as WO2011 / 120053, same as WO2011 / 100131, same as WO2011 / 163121, same as WO2012 / 177947, same as WO2013 / 033230, same as W No. O2013 / 075035, same as No. WO2012 / 083185, same as No. WO2012 / 083046, same as No. WO2009 / 082607, same as No. WO2009 / 134487, same as No. WO2010 / 144740, same as No. WO2010 / 148013, same as No. WO1997 / 020563, same as No. WO2010 / 088537, same as No. WO2002 / 043771, same as No. WO2010 / 129709, same as No. WO2012 / 068187, same as No. WO2009 / 126933, same as No. WO2004 / 024757 No., same as WO2010 / 054406, same as WO2012 / 089352, same as WO2012 / 089602, same as WO2013 / 166121, same as WO2013 / 165816, US Patent No. 4,751,219, same as 7,582,744, Same as No. 8,552,163, same as No. 8,137,695, same as No. 6,908,903, same as No. 6,383,812, same as No. 7,26 No. 2,177, same as No. 6,525,031, same as No. 5,994,517, same as No. 6,660,720, same as No. 6,300,319 , Same as No. 7,723,509, Same as No. 8,106,022, Same as No. 7,491,805, Same as No. 7,491,805, Same as No. 8,5 No. 41,548, same as No. 8,344,125, same as No. 8,313,772, same as No. 8,349,308, same as No. 8,450,467 No. 8,501,930, 8,158,601, 7,262,177, 6,906,182, 6,620,916, 8,435,491, 8,404,862, 7,851,615, U.S. Patent Application No. U.S.2011 / 0097264, US2011 / 0097265, US2013 / 0004427, US2003 / 0119724, US2011 / 0207799, US2012 / 0035115, U S2012 / 0230938, US2005 / 0164235, US2006 / 0183886, US2012 / 0136042, US2012 / 0095075, US2013 / 0109817, US Examples of linking groups described in references US2006 / 0148740, US2008 / 0206869, US2012 / 0165393, US2012 / 0101148, US2013 / 0121954, US2011 / 0123520, US2003 / 0077829, US2008 / 0108801, and US2009 / 0203132 are, but are not limited to, all of which are incorporated herein by reference.

[0272] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] The formula includes or collectively includes a structure selected from, where each n is an integer between 1 and 20 (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20), and p is an integer between 1 and 6 (i.e., 1, 2, 3, 4, 5, or 6).

[0273] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] The expression includes a structure selected from or includes such a structure as a whole, where each n is an integer independently between 1 and 20 (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20).

[0274] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] The expression includes a structure selected from or includes such a structure as a whole, where each n is an integer independently between 1 and 20 (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20).

[0275] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] The expression includes a structure selected from or includes such a structure as a whole, where each n is an integer independently between 1 and 20 (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20).

[0276] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] The formula includes or collectively includes a structure selected from the following, where each L is independently a phosphorus linking group, and each n is independently an integer between 1 and 20 (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20).

[0277] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] [ka] It includes a structure selected from, or includes that structure as an integral part of.

[0278] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] It includes a structure selected from, or includes that structure as an integral part of.

[0279] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] It includes a structure selected from, or includes that structure as an integral part of.

[0280] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] The formula includes or incorporates a structure selected from the following, where n is 1 to 20 (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20).

[0281] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] It includes a structure selected from, or includes that structure as an integral part of.

[0282] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] It includes a structure selected from, or includes that structure as an integral part of.

[0283] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] It includes a structure selected from, or includes that structure as an integral part of.

[0284] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] It includes or integrally has the structure of

[0285] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] It includes or integrally has the structure of

[0286] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] It includes a structure selected from, or includes that structure as an integral part of.

[0287] In certain embodiments, L1, L2, L3, and L4 are independent of each other. [ka] The formula includes a structure selected from or includes such a structure as a whole, where each n is independently 0, 1, 2, 3, 4, 5, 6, or 7.

[0288] In some embodiments, L1, L2, L3, and L4 may independently be a linker (e.g., an optionally substituted alkyl linker, an optionally substituted polyethylene glycol (PEG) linker, an optionally substituted heteroalkyl linker, or an optionally substituted heteroaryl linker). In some embodiments, L1, L2, L3, and L4 may independently be a bond (e.g., an intercarbon bond, a phosphodiester bond, or a phosphorothioate bond). In some embodiments, L1, L2, L3, and L4 may independently be absent.

[0289] In some embodiments, L1 is a bond. In some embodiments, L1 is an optionally substituted alkyl linker. In some embodiments, L1 is an optionally substituted C1-C6 alkyl linker. In some embodiments, L1 is a C1-C6 alkyl linker substituted with =O. In certain embodiments, L1 is [ka] It includes the structure.

[0290] In some embodiments, L2 is an optionally substituted alkyl linker. In some embodiments, L2 is an optionally substituted C1-C 15 It is an alkyl linker. In some embodiments, L2 is optionally substituted C5-C 12 It is an alkyl linker. In certain embodiments, L2 is [ka] The structure includes, [ka] It includes the structure.

[0291] In some embodiments, L2 is an optionally substituted PEG linker. In some embodiments, L2 is an optionally substituted PEG linker whose PEG unit length is 1, 2, 3, 4, 5, 6, 7, or 8, and whose PEG unit is [ka] The structure includes: In certain embodiments, L2 is an optionally substituted PEG linker with a PEG unit length of 3. In certain embodiments, L2 is an optionally substituted PEG linker with a PEG unit length of 4. In certain embodiments, L2 is [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] The structure includes, [ka] It includes the structure.

[0292] In some embodiments, L2 is an optionally substituted heteroalkyl linker. In certain embodiments, L2 is [ka] The structure includes, [ka] It includes the structure.

[0293] In some embodiments, L3 is an optionally substituted heteroaryl linker. In some embodiments, L3 is an optionally substituted partially unsaturated heterocycloalkyl linker or heteroaryl linker. In certain embodiments, L3 is [ka] It includes the structure.

[0294] In some embodiments, L4 is an optionally substituted heteroalkyl linker. In some embodiments, the heteroalkyl linker is substituted with one or more =O substituents. In certain embodiments, L4 is [ka] The structure includes, where X is O or S. In a particular embodiment, L4 is [ka] The structure includes the form where X is either O or S.

[0295] In some embodiments, L1, L2, L3, and L4 are integrated as a single unit. [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes, where X is O or S. In some embodiments, L1, L2, L3 and L4 are integrated as follows: [ka] The structure includes L1, L2, L3 and L4 together. [ka] The structure includes the form where X is either O or S.

[0296] Method for preparing compounds In some embodiments, this disclosure relates to methods for preparing compounds and compositions containing NMDA receptor ligands as disclosed herein.

[0297] The compounds of this disclosure can be prepared by organic synthesis methods known in the art. Where necessary, methods for optimizing reaction conditions and minimizing competing byproducts are known in the art. Conveniently, reaction optimization and scale-up may be performed using high-speed parallel synthesis apparatuses and computer-controlled microreactors (e.g., Design and Optimization in Organic Synthesis, 2nd Edition, Carlson R, Ed, 2005; Elsevier Science Ltd., Jahnisch, K et al., Angew. Chem. Int. Ed. Engl. 2004 43:406 and references herein). Additional reaction schemes and reaction protocols may be determined by those skilled in the art using commercially available structure-searchable database software, e.g., SciFinder® (CAS Division of the American Chemical Society) and Reaxys® (Elsevier), or by using an internet search engine, e.g., Google®, or by appropriate keyword searches using keyword databases, e.g., the U.S. Patent and Trademark Office text database.

[0298] As will be apparent to those skilled in the art, the method for synthesizing the compounds of the formulas herein, including the schemes and examples herein, will be clear to them. In addition, various synthesis steps may be carried out in different orders or sequences to obtain the desired compounds. Furthermore, the solvents, temperatures, reaction periods, etc., disclosed herein are for illustrative purposes only, and those skilled in the art will recognize that variations in these reaction conditions can produce the desired compounds of this disclosure.

[0299] The compounds of the present invention may include bonds whose rotation is restricted with respect to a particular bond (e.g., carbon-carbon bonds), for example, due to the presence of a ring or double bond. Therefore, it is clear that all cis / trans and E / Z isomers are included in this disclosure. The compounds of the present invention may also be represented in numerous tautomers, in which case it is clear that all tautomers of the compounds described herein are included, even if only one tautomer is represented. It is clear that all such isomers of such compounds of the present invention are included in this disclosure. It is clear that all crystalline forms and crystalline polymorphs of the compounds described herein are included in this disclosure. Extracts and fractions containing the compounds of this disclosure are also embodied. The term “isomer” is intended to include diastereomers, enantiomers, positional isomers, structural isomers, rotational isomers, tautomers, etc. For compounds containing one or more stereocenters, such as chiral compounds, the methods of this disclosure may be carried out with enantiomerically enriched compounds, racemic compounds, or diastereomer mixtures. It is clear that all isomers of the compounds revealed herein are included in this disclosure.

[0300] A preferred enantiomerically enriched compound has an enantiomer excess of 50% or more. More preferably, the compound has an enantiomer excess of 60%, 70%, 80%, 90%, 95%, 98%, or 99% or more. In a preferred embodiment, only one of the enantiomers or diastereomers of the chiral compound of the Disclosure is administered to cells or subjects.

[0301] Treatment method In one embodiment, the present invention provides a method for treating a subject who is suffering from or susceptible to a disorder or disease, comprising administering to the subject an effective amount of a compound or pharmaceutical composition described herein.

[0302] In other embodiments, the present invention provides a method for treating a subject that is suffering from or susceptible to a disorder or disease, wherein the subject is identified as requiring modification of protein function, and the method comprises administering an effective amount of a compound or pharmaceutical composition described herein to the subject requiring such modification to treat the disorder in the subject.

[0303] In one embodiment, the provided method for delivering a therapeutic oligonucleotide to a target brain comprises contacting the target with a compound or pharmaceutical composition described herein in an amount and under conditions that target the brain. In some embodiments, the therapeutic oligonucleotide is delivered to one or more brain regions selected from the group consisting of the striatum, cerebellum, brainstem, hippocampus, prefrontal cortex, and spinal cord.

[0304] In certain embodiments, the invention provides a method for treating a disease, disorder, or symptom thereof, wherein the disease is a disease, disorder, or symptom thereof of the central nervous system (CNS). In some embodiments, the disease is a neurodegenerative disease, disorder, or symptom thereof. In some embodiments, the disease is Alzheimer's disease or a symptom thereof.

[0305] Examples of CNS disorders include, but are not limited to, neurotoxicity and / or neurotrauma, stroke, multiple sclerosis, spinal cord injury, epilepsy, psychiatric disorders, sleep disorders, motor disorders, nausea and / or vomiting, amyotrophic lateral sclerosis, Alzheimer's disease, and substance abuse or substance use disorder (SUD).

[0306] In certain embodiments, the CNS impairment is due to neurotoxicity and / or neurotrauma, for example, acute neuronal injury (e.g., traumatic brain injury (TBI), stroke, epilepsy), or chronic neurodegenerative disorders (e.g., multiple sclerosis, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, Alzheimer's disease). In certain embodiments, the compounds of this disclosure provide, for example, neuroprotective effects against acute neuronal injury or chronic neurodegenerative disorders.

[0307] In certain embodiments, the CNS disorder is a stroke (e.g., ischemic stroke).

[0308] In certain embodiments, the CNS disorder is multiple sclerosis.

[0309] In certain embodiments, the CNS impairment is spinal cord injury.

[0310] In certain embodiments, the CNS disorder is epilepsy.

[0311] In certain embodiments, the CNS disorder is a mental disorder, such as depression, anxiety or anxiety-related conditions, learning disabilities, somatic symptom disorders, schizophrenia, or schizoaffective disorder.

[0312] In certain embodiments, the CNS disorder is depression. "Depression" includes, but is not limited to, depressive disorders or conditions, such as major depressive disorder (e.g., unipolar depression), treatment-resistant depression, dysthymic disorder (e.g., chronic mild depression), bipolar disorder (e.g., manic-depressive illness), seasonal affective disorder, and / or depression associated with substance abuse or substance abuse disorder (e.g., withdrawal symptoms). The depression may be clinical or subclinical. The depression may be associated with premenstrual syndrome and / or premenstrual dysphoric disorder, or may be premenstrual syndrome and / or premenstrual dysphoric disorder.

[0313] In certain embodiments, the CNS disorder is anxiety. "Anxiety" includes, but is not limited to, anxiety and anxiety-related conditions, such as clinical anxiety, panic disorder, agoraphobia, generalized anxiety disorder (GAD), specific phobias, social phobias, obsessive-compulsive disorder, acute stress disorder, post-traumatic stress disorder, adjustment disorder with anxiety features, anxiety disorders associated with depression, anxiety disorders due to systemic medical conditions, substance-induced anxiety disorders, anxiety associated with substance abuse or substance use disorder (e.g., withdrawal symptoms, dependence, recovery), and anxiety associated with nausea and / or vomiting. The treatment may also be aimed at inducing or promoting sleep in the subject (e.g., the anxious subject).

[0314] In certain embodiments, the CNS disorder is a learning disability (e.g., attention deficit disorder (ADD)).

[0315] In certain embodiments, the CNS disorder is schizophrenia or schizoaffective disorder.

[0316] In certain embodiments, the CNS disorder is a sleep disorder. "Sleep disorders" include, but are not limited to, insomnia, narcolepsy, sleep apnea, restless legs syndrome (RLS), delayed sleep phase syndrome (DSPS), periodic limb movement disorder (PLMD), hypopnea syndrome, rapid eye movement disorder (RBD), shift work sleep disorder (SWSD), and sleep problems (e.g., abnormal sleep behavior), such as nightmares, night terrors, sleep talking, head thuds, snoring, and teeth clenching and / or grinding (bruxism).

[0317] In certain embodiments, the CNS disorder is a motor disorder, such as a basal ganglia disorder, such as Parkinson's disease, levodopa-induced dyskinesia, Huntington's disease, Tourette syndrome, tardive dyskinesia, and dystonia.

[0318] In certain embodiments, the CNS disorder is Alzheimer's disease.

[0319] In certain embodiments, the CNS disorder is amyotrophic lateral sclerosis (ALS).

[0320] In certain embodiments, the CNS disorder is nausea and / or vomiting.

[0321] In certain embodiments, the CNS disorder is a substance abuse or substance use disorder (SUD) (e.g., addiction to opioids, nicotine, cocaine, psychostimulants, or alcohol).

[0322] The term “neurological disorder” (including, for example, “neurodegenerative disorder”) refers to any disorder of the nervous system, including disorders involving the central nervous system (brain, brainstem, and cerebellum), the peripheral nervous system (including cranial nerves), and the autonomic nervous system (some of which are present in both the central and peripheral nervous systems). Neurodegenerative disorder refers to a type of neurological disorder characterized by the loss of nerve cells, and includes, but is not limited to, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, tauopathies (including frontotemporal dementia), and Huntington’s disease. Examples of neurological disorders include, but are not limited to, headaches, stupor and coma, dementia, seizures, sleep disorders, trauma, infections, neoplasms, neuro-ocular disorders, motor disorders, demyelinating disorders, spinal cord disorders, and disorders of the peripheral nerves, muscles, and neuromuscular junctions. Substance abuse or use disorders (SUDs) and mental disorders (including, but not limited to, bipolar disorder, schizophrenia, and schizoaffective disorder) are also included in the definition of neurological disorders. Further examples of neurological disorders include acquired epileptic aphasia, acute disseminated encephalomyelitis, adrenoleukodystrophy, corpus callosum agenesis, agnosia, Ecardi syndrome, Alexander disease, Alpers disease, alternating hemiplegia, Alzheimer's disease, amyotrophic lateral sclerosis, anencephaly, Angelman syndrome, hemangioma, oxygen deficiency, aphasia, apraxia, arachnoid cyst, arachnoiditis, Arnold-Chiari malformation, arteriovenous malformation, Asperger's syndrome, telangiectasia ataxia, attention deficit hyperactivity disorder, autism, autonomic dysfunction, back pain, Batten's disease, Behçet's disease, Bell's palsy, benign essential blepharospasm, benign focal muscular atrophy, benign intracranial hypertension, and Binswanger's syndrome. Diseases, blepharospasm, Bloch-Salzberger syndrome, brachial plexus injury, brain abscess, brain injury, brain tumor (including glioblastoma multiforme), spinal cord tumor, Brown-Séquard syndrome, Canavan disease, carpal tunnel syndrome (CTS), causalgia, central pain syndrome, central pontine myelinlysis, head injury, cerebral aneurysm, cerebral arteriosclerosis, cerebral atrophy, cerebral gigantism, cerebral palsy, Charcot-Marie-Tooth disease, chemotherapy-induced neuropathy and neuropathic pain, Chiari malformation, chorea, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic pain, chronic focal pain syndrome, Coffin-Lowry syndrome, coma (including persistent vegetative state), congenital bilateral facial nerve palsy, corticobasal degeneration,Cerebral arteritis, craniosynostosis, Creutzfeldt-Jakob disease, accumulative trauma, Cushing's syndrome, giant cell inclusion body disease (CIBD), cytomegalovirus infection, Dancing Eyes-Dancing Feet syndrome, Dandy-Walker syndrome, Dawson's disease, Demorsia syndrome, Krampke palsy, dementia, dermatomyositis, diabetic neuropathy, pervasive sclerosis, autonomic neuropathy, dysgraphia, dyslexia, dystonia, early infantile epileptic encephalopathy, sella turcica syndrome, encephalitis, brain herniation, trigeminal nerve hemangioma, epilepsy, Erb's palsy, essential tremor, Fabry disease, Fahl Diseases, syncope, familial spastic paralysis, febrile seizures, Fisher syndrome, Friedreich's ataxia, frontotemporal dementia and other "tauopathies", Gaucher disease, Gerstmann syndrome, giant cell arteritis, giant cell inclusion disease, globular cell leukodystrophy, Guillain-Barré syndrome, HTLV-1-associated myelopathy, Harerforden-Spats disease, head injury, headache, hemifacial spasm, hereditary spastic paraplegia, hereditary polyneurotic ataxia, herpes zoster, herpes zoster, Hirayama syndrome, HIV-related dementia and neurological disorders (see also neurological signs of AIDS), holoprosencephalopathy, Huntington's disease and others Polyglutamine repeat disease, anencephaly, hydrocephalus, hyperadrenocorticism, hypoxia, immune-mediated encephalomyelitis, inclusion body myositis, incontinentia pigmenti, infantile phytanate storage disease, infantile Refsum disease, infantile seizures, inflammatory muscle disease, intracranial cysts, increased intracranial pressure, Joubert syndrome, Keens-Sayer syndrome, Kennedy disease, Kinsborne syndrome, Klippel-Feil syndrome, Krabbe disease, Kugelberg-Wellander disease, Kuru, Lafora disease, Lambert-Eaton myasthenic syndrome, Landau-Klefner syndrome, lateral medullary (Wallenberg) syndrome, learning disability, Leigh disease, Leigh Knox-Gastaut syndrome, Lesch-Nyhan syndrome, leukodystrophy, Lewy body dementia, lissencephaly, locked-in syndrome, Lou Gehrig's disease (also known as motor neuron disease or amyotrophic lateral sclerosis), lumbar disc disease, Lyme disease (neurological sequelae), Machado-Joseph disease, cerebral encephalopathy, megacephaly, Melkerson-Rosenthal syndrome, Meniere's disease, meningitis, Menkes disease, metachromatic leukodystrophy, microcephaly, migraine, Miller-Fischer syndrome, ministroke, mitochondrial myopathy, Möbius syndrome, unilateral upper limb muscular atrophy,Motor neuron disease, Moyamoya disease, Mucopolysaccharidosis, Multiple infarct dementia, Multifocal motor neuropathy, Multiple sclerosis and other demyelinating disorders, Multiple system atrophy with orthostatic hypotension, Muscular dystrophy, Myasthenia gravis, Myelin-disintegrating generalized sclerosis, Myoclonic encephalopathy in infants, Myoclonus, Myopathy, Congenital myotonia, Narcolepsy, Neurofibromatosis, Neuroleptic malignant syndrome, Neurological signs of AIDS, Neurological sequelae of lupus, Neuromyotonia, Neuronal ceroid lipofuscinosis, Neuronal cell migration disorders, Niemann-Pick disease, O'Sullivan-McLeod syndrome, Occipital nerve Pain, secondary occult spinal insufficiency, Ohtahara syndrome, olivopontocerebellar atrophy, opsoclonus-myoclonus, optic neuritis, orthostatic hypotension, overuse syndrome, paresthesia, Parkinson's disease, congenital paramyotonia, paraneoplastic cerebellar disease, paroxysmal seizures, Parry-Romberg syndrome, Pelizaeus-Merzbacher disease, periodic paralysis, peripheral neuropathy, painful neuropathy and neuropathic pain, persistent vegetative state, pervasive developmental disorder, photic sneeze reflex, phytanic acid storage, Pick's disease, nerve compression, pituitary tumor, polymyositis, porencephaly, post-polio syndrome, postherpetic neuralgia (PH N) Post-infectious encephalomyelitis, orthostatic hypotension, Prader-Willi syndrome, primary lateral sclerosis, prion disease, progressive hemifacial atrophy, progressive multifocal leukoencephalopathy, progressive sclerosing poliodystrophy, progressive supranuclear palsy, pseudotumor, Ramsay Hunt syndrome (types I and II), Rasmussen encephalitis, reflex sympathetic dystrophy, Refsum disease, repetitive movement disorder, repetitive hyperinjury, restless legs syndrome, retrovirus-associated myelopathy, Rett syndrome, Reye's syndrome, St. Vitus' Dance, Sandhoff disease, Schilder's disease, schizencephaly, septal-optic nerve malformation, shaken baby Child syndrome, herpes zoster, Shy-Drager syndrome, Sjögren's syndrome, sleep apnea, Sotos syndrome, spasticity, spina bifida, spinal cord injury, spinal cord tumor, spinal muscular atrophy, stiff person syndrome, stroke, Sturge-Weber syndrome, subacute sclerosing panencephalitis, subarachnoid hemorrhage, subcortical arteriosclerotic encephalopathy, Sydenham's chorea, syncope, syringomyelia, tardive dyskinesia, Tay-Sachs disease, temporal arteritis, tethered cord syndrome, Thomsen's disease, thoracic outlet syndrome, painful tics, Todd's palsy, Tourette's syndrome, transient ischemic attack, transmissible spongiform encephalopathy, transverse myelitis, traumatic brain injury,These include tremor, trigeminal neuralgia, tropical spastic paraplegia, tuberous sclerosis, vascular dementia (multiple infarct dementia), vasculitis (including temporal arteritis), von Hippel-Lindau disease (VHL), Wallenberg syndrome, Werdnig-Hoffmann disease, West syndrome, whiplash, Williams syndrome, Wilson's disease, and Zellweger syndrome.

[0323] In certain embodiments, the subject is a mammal, preferably a primate or a human.

[0324] In another embodiment, the method provided is as described above, and the effective amount of the compound provided in the present invention is as described above.

[0325] In another embodiment, the method provided is as described above, and the compound provided in the present invention is administered intrathecal, intravenously, intramuscularly, subcutaneously, intracerebroventricularly, orally, or topically. In a particular embodiment, the compound is administered intrathecally.

[0326] In other embodiments, the method provided is as described above, in which a compound of any of the formulas shown herein is administered alone or in combination with one or more other pharmaceuticals. In further embodiments, the additional therapeutic agent is a central nervous system (CNS) disease agent.

[0327] Another purpose of this disclosure is to use compounds such as those described herein in the manufacture of pharmaceuticals for use in the treatment of disorders or diseases.

[0328] Pharmaceutical composition In one embodiment, the provided material is a pharmaceutical composition comprising any of the compounds described herein and a pharmaceutically acceptable carrier or a pharmaceutically acceptable excipient.

[0329] Compounds or compositions as described herein can be administered in combination with one or more additional therapeutic agents (e.g., therapeutically and / or prophylactically active agents). The compound or composition can be administered in combination with additional therapeutic agents that improve the activity (e.g., activity (e.g., potency and / or efficacy), improve bioavailability, improve safety, reduce drug resistance, reduce and / or modify metabolism), inhibit excretion, and / or modify distribution in a subject or cell, in order to treat a disease in a subject requiring treatment, prevent a disease in a subject requiring prevention, and / or reduce the risk of disease development in a subject requiring reduction of that risk. It will also be apparent that the therapies used may produce the desired effect on the same disorder, and / or produce different effects. In certain embodiments, a pharmaceutical composition as described herein, comprising a compound and an additional therapeutic agent, exhibits synergistic effects not present in pharmaceutical compositions comprising either the compound or one of the additional therapeutic agents described herein, but not both.

[0330] The compound or composition may be administered concurrently with one or more additional therapeutic agents, either before or after them, the therapeutic agents which may be useful, for example, as combination therapy agents. Examples of therapeutic agents include therapeutic activators. Examples of therapeutic agents include prophylactic activators. Examples of therapeutic agents include small organic molecules, such as drug compounds (e.g., compounds approved by the U.S. Food and Drug Administration for use in humans or livestock, as shown in the Code of Federal Regulations (CFR)), peptides, proteins, glycans, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or synthetic proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNA, RNA, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells. In certain embodiments, the additional therapeutic agent is a therapeutic agent useful for treating and / or preventing a disease (e.g., CNS disorders). Each additional therapeutic agent may be administered in doses and / or time schedules determined for that therapeutic agent. The additional therapeutic agents may be administered together with each other and / or with the compounds or compositions described herein in a single dose, or separately in different doses. In any particular combination used in a regimen, the compatibility of the compounds described herein with the additional therapeutic agent(s), and / or the desired therapeutic and / or preventive effects to be achieved should be considered. Generally, the additional therapeutic agent(s) in a combination are expected to be used at levels no higher than those used individually. In some embodiments, the levels used in combination are lower than those used individually.

[0331] In one embodiment, the provided kit contains an effective amount of the compound provided in the present invention in unit dosage form, along with instructions for administering the compound to a person suffering from or susceptible to a disease or disorder.

[0332] The terms “pharmaceutically acceptable salt” or “pharmaceutically acceptable carrier” are intended to include salts of the active compound that, depending on the specific substituents found in the compounds described herein, are prepared with relatively non-toxic acids or bases. When the compounds of this disclosure contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of the compound with a sufficient amount of the desired base, either in its original form or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include salts of sodium, potassium, calcium, ammonium, organic amino, or magnesium, or similar salts. When the compounds of this disclosure contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of the compound with a sufficient amount of the desired acid, either in its original form or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts derived from inorganic acids (such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monocarbonate, phosphoric acid, monohydrogen phosphoric acid, dihydrogen phosphoric acid, sulfuric acid, monohydrosulfuric acid, hydroiodic acid, or phosphorous acid), and salts derived from relatively non-toxic organic acids (such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid). Salts of amino acids, such as alginates, and salts of organic acids (such as glucuronic acid or galactunolonic acid) are also included (see, for example, Berge et al., Journal of Pharmaceutical Science 66:1-19 (1977)). The specific compounds of this disclosure contain both basic and acidic functional groups that enable the compound to be converted into either a base addition salt or an acid addition salt. Other pharmaceutically acceptable carriers known to those skilled in the art are suitable for this disclosure.

[0333] The neutral form of the compound may be regenerated by contacting its salt with a base or acid to isolate the parent compound in its conventional form. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but apart from these salts, for the purposes of this disclosure, they are equivalent to the parent form of the compound.

[0334] This disclosure provides compounds in prodrug form in addition to salt form. The prodrugs of the compounds described herein are compounds that readily undergo chemical transformation under physiological conditions to yield the compounds of this disclosure. In addition, the prodrugs can be converted to the compounds of this disclosure in an ex vivo environment by chemical or biochemical methods. For example, the prodrug can be slowly converted to the compounds of this disclosure when placed in a reservoir-type transdermal patch with appropriate enzymes or chemical reagents.

[0335] Certain compounds in this disclosure may exist in non-solvated and solvated (including hydrated) forms. Generally, their solvated forms are equivalent to and intended to be within the scope of this disclosure. Certain compounds in this disclosure may exist in numerous crystalline or amorphous forms. Generally, all physical forms are equivalent for the applications envisioned by this disclosure and are intended to be within the scope of this disclosure.

[0336] This disclosure also provides pharmaceutical compositions comprising an effective amount of the compounds described herein and pharmaceutically acceptable excipients. In one embodiment, any compound of any of the formulas shown herein is administered to a subject using a pharmaceutically acceptable formulation, for example, a pharmaceutically acceptable formulation that delivers the compound continuously for at least 12 hours, 24 hours, 36 hours, 48 ​​hours, 1 week, 2 weeks, 3 weeks, or 4 weeks after the pharmaceutically acceptable formulation is administered to the subject.

[0337] The actual dose level and administration time course of the active ingredient in the pharmaceutical compositions of this disclosure may be varied to obtain an amount of the active ingredient that is effective in achieving a desired therapeutic response in a particular subject, composition, and administration method, while also being tolerable for that subject.

[0338] When used, at least one compound of the present disclosure is administered in a pharmaceutically effective amount to the subject requiring administration, in a pharmaceutical carrier, by intravenous injection, intrathecal injection, intramuscular injection, subcutaneous injection or intracerebroventricular injection, or by oral administration or topical application. The compounds of the present disclosure may be administered alone or in combination with a second different pharmaceutical product, where “in combination with” means together, substantially simultaneously or sequentially. In one embodiment, the compounds of the present disclosure are administered over a short period; that is, the compounds of the present disclosure may be administered for a short treatment period, e.g., about one day to about one week. In another embodiment, the compounds of the present disclosure may be administered over a long period, e.g., about one week to several months, depending on the condition being treated, to improve a chronic disorder.

[0339] "Pharmacologically effective dose," as used herein, means a dose of the compound disclosed that, within reasonable medical judgment, is sufficiently high to significantly and positively modify the condition being treated, but sufficiently low to avoid serious side effects (in a reasonable benefit / risk ratio). The pharmaceutically effective dose of the compound disclosed will vary depending on the specific treatment objective, the age and physical condition of the patient being treated, the severity of the underlying disease, the duration of treatment, the nature of the concomitant therapy, and the specific compound used. For example, the therapeutically effective dose of the compound disclosed for administration to children or neonates will decrease proportionally according to reasonable medical judgment. That is, the effective dose of the compound disclosed will be the minimum dose that produces the desired effect.

[0340] A key practical advantage of the present disclosure is that the compounds may be administered in conventional forms, for example, by intrathecal, intravenous, intramuscular, subcutaneous, oral, or intracerebroventricular injection routes, or by topical application such as creams or gels. Depending on the route of administration, the active ingredients containing the compounds of the present disclosure may need to be coated with a substance to protect the compound from the action of enzymes, acids, and other natural conditions that may inactivate it. For administration of the compounds of the present disclosure by means other than parenteral administration, the compounds may be coated with a substance to prevent inactivation, or the compounds may be administered together with such a substance.

[0341] The compound may be administered parenterally or intraperitoneally. Dispersions can also be prepared, for example, with glycerol, liquid polyethylene glycol, and mixtures thereof, as well as oils.

[0342] Some examples of substances that can function as pharmaceutical excipients or pharmaceutical carriers (these terms are used synonymously herein) include, for example, 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; powdered tragacanth; malt; gelatin; talc; stearic acid; magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and theobroma oil; polyols such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; agar; alginic acid; pyrogen-free water; isotonic saline and phosphate buffer; skim milk powder; and other non-toxic, suitable substances for use in pharmaceutical formulations, such as vitamin C, estrogen, and echinacea. Wetting agents and lubricants, such as sodium lauryl sulfate, as well as colorants, flavoring agents, lubricants, excipients, tableting agents, stabilizers, antioxidants, and preservatives may also be present. Solubilizers (including, for example, cremophor and β-cyclodextrin) can also be used in the pharmaceutical compositions of the present invention.

[0343] Pharmaceutical compositions comprising the active compounds of this disclosure (or their derivatives or prodrugs) can be prepared by conventional mixing, dissolution, granulation, sugar-coated tablet preparation, polishing, emulsification, encapsulation, encapsulation, or lyophilization processes. The compositions can be prepared conventionally using one or more physiologically acceptable carriers, diluents, excipients, or auxiliaries that facilitate the processing of the active compounds into pharmaceutically usable preparations. The compositions of the present invention can be prepared (for example, for pharmaceutical, agricultural, or livestock use) by combining the compounds disclosed herein with one or more suitable carriers, diluents, excipients, or auxiliaries (including those described herein) (for example, by contacting, mixing, dissolving, granulating, sugar-coated tablet preparation, polishing, emulsifying, encapsulating, encapsulating, or lyophilizing).

[0344] The pharmaceutical compositions of this disclosure may take a form substantially suitable for any method of administration (including, for example, intrathecal administration, topical administration, ophthalmic administration, oral administration, systemic administration, nasal administration, injection administration, transdermal administration, rectal administration, vaginal administration, etc.), or a form suitable for administration by inhalation or inhalation.

[0345] Systemic formulations include formulations designed for administration by injection, such as subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection, as well as formulations designed for transdermal, transmucosal, oral, or pulmonary administration.

[0346] Useful injectable preparations include sterile suspensions, solutions, or emulsions of the active compound(s) of the present invention in an aqueous or oily vehicle. The composition may also include compounding agents, such as suspending agents, stabilizers, and / or dispersants. Injectable formulations may be provided in unit dosage forms (e.g., ampoules or multi-dose containers) and may include added preservatives.

[0347] Alternatively, the injectable formulation may be provided in powder form for reconstitution with a suitable vehicle before use, such as, but not limited to, sterile pyrogen-free water, buffers, or dextrose solutions. For this purpose, the active compound(s) of the present invention may be dried by any technique known in the art, for example, lyophilization, and reconstituted before use.

[0348] For sustained delivery, the active compound(s) or prodrug(s) can be formulated as a depot preparation for implantation or intramuscular injection. The active component can be formulated with a suitable polymer or hydrophobic substance (e.g., as an emulsion in an acceptable oil), or with an ion exchange resin, or as a sparingly soluble derivative, such as a sparingly soluble salt.

[0349] Alternatively, other drug delivery systems can be used. Liposomes and emulsions are well-known examples of delivery vehicles that can be used to deliver active compounds, oligonucleotides, or prodrugs. Certain organic solvents, such as dimethyl sulfoxide (DMSO), can also be used.

[0350] The pharmaceutical composition of the present invention may be provided in a pack or dispenser device that may contain, if desired, one or more unit dosage forms comprising the active compound. The pack may include, for example, metal or plastic foil, such as a blister pack. Instructions for administration may be attached to the pack or dispenser device.

[0351] The active compounds(s) or prodrugs(s) or compositions thereof disclosed herein are generally to be used in amounts effective to achieve the intended effect, for example, in amounts effective to treat or prevent the specific disease being treated. The compounds(s) and oligonucleotides(s) may be administered therapeutically to achieve a therapeutic benefit or prophylactically to achieve a preventive benefit. A therapeutic benefit means eliminating or improving the underlying disorder being treated and / or eliminating or improving one or more symptoms associated with the underlying disorder so that the patient reports an improvement in sensation or condition, even if the patient still suffers from the underlying disorder. A therapeutic benefit also includes stopping or slowing the progression of the disease, whether or not improvement is achieved.

[0352] In prophylactic administration, the compound can be administered to patients at risk of developing one of the aforementioned diseases. Patients at risk of developing a disease may be those who possess characteristics that define them as belonging to a group identified as at risk, as defined by the appropriate healthcare professional or healthcare group. Patients at risk may also be those who are generally or typically exposed to environments in which the underlying disease is likely to manifest. In other words, patients at risk are those who are generally or typically exposed to the disease or illness causing the condition, or who may be exposed for a limited period of time or for a short duration. Alternatively, prophylactic administration may be applied to patients diagnosed with the underlying disorder to prevent the onset of symptoms.

[0353] The dosage of a compound depends on various factors, including, for example, the specific indication being treated, the method of administration, whether the desired benefit is preventive or therapeutic, the severity of the indication being treated, the patient's age and weight, and the bioavailability of the specific active compound. Determining the effective dosage is well within the capabilities of those skilled in the art.

[0354] The effective dose can be initially estimated from in vitro assays. For example, the initial dose for use in animals is the concentration of the active compound in circulating blood or serum, as measured by in vitro assays, e.g., the MIC or MFC of fungi in vitro, and other in vitro assays, and the IC of that particular compound. 50 The formulation can be adjusted to achieve concentrations such as those listed above. Calculating the dosage to achieve such circulating blood or serum concentrations, while considering the bioavailability of the particular compound, is well within the capabilities of those skilled in the art. For guidance, please refer to “General Principles,” In: Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, Chapter 1, pp. 1-112, 13th ed., McGraw-Hill, and the references thereto, which are incorporated herein by reference.

[0355] Initial doses can also be estimated from in vivo data, such as animal models. Animal models useful for testing the efficacy of compounds to treat or prevent the various diseases mentioned above are well known in the art.

[0356] Dosages typically range from approximately 0.0001, 0.001, or 0.01 mg / kg / day to approximately 100 mg / kg / day, but can be higher or lower depending on several factors, particularly the compound's activity, bioavailability, administration method, and the various factors mentioned above. Dosage and intervals can be individually adjusted to achieve plasma levels of the compound(s) sufficient to maintain a therapeutic or prophylactic effect. In the case of local or selective administration, for example, local topical administration, the effective local concentration of the active compound(s) is not limited to plasma concentrations. Those skilled in the art will be able to optimize the effective local dose without excessive experimentation.

[0357] Preferably, the compound(s) of the present invention will provide therapeutic or preventive benefits and will be tolerably accepted. The tolerability of the compound(s) and oligonucleotide(s) can be determined using standard pharmaceutical procedures. The dose ratio between the unacceptable effect and the therapeutic (or preventive) effect is the therapeutic index. Compound(s) with a high therapeutic index are preferred.

[0358] Where a list of chemical groups is enumerated in any definition of a variable in this specification, the definition of that variable is included as any one of the enumerated groups or in combination thereof. Where embodiments of a variable are enumerated in this specification, the embodiments are included as any one of the individual embodiments or in combination with any other embodiment or part thereof. Where embodiments are enumerated in this specification, the embodiments are included as any one of the individual embodiments or in combination with any other embodiment or part thereof. [Examples]

[0359] The following examples are provided to allow for a deeper understanding of the embodiments described herein. The examples described herein are provided to illustrate the compounds, compositions and methods described herein and should not be construed as limiting their scope in any way.

[0360] Common synthesis routes The compound of formula (I) can be prepared according to the following general synthetic route. [ka]

[0361] I. Preparation of the ligand synthesis intermediate AY-L1-L2-N3, in which Y is -C(=O)- The ligand-linker-azide synthetic intermediate of general formula AY-L1-L2-N3 can be prepared according to one of the general procedures outlined below.

[0362] In some cases, NMDA receptor ligand A contains an amino group coupled to an acidic group of a linker, the linker further containing an azide. In some cases, the amine is a primary amine. In other cases, the amine is a secondary amine forming part of the ring. In some cases, the coupling is achieved using an activating group, such as a carbodiimide-mediated modification of a carboxylic acid (e.g., N-hydroxysuccinimide).

[0363] In other cases, NMDA receptor ligand A contains an acidic group coupled to an amine group of a linker, and the linker further contains an azide. [ka]

[0364] II. Preparation of the ligand synthesis intermediate AY-L1-L2-N3, in which Y is a bond. In some cases, NMDA receptor ligand A comprises an amino group coupled to a linker containing a leaving group (e.g., Br, Cl, I, OMs, etc.), the linker further comprising an azide. In other cases, the linker comprises an amino group coupled to NMDA receptor ligand A containing a leaving group (e.g., Br, Cl, I, OMs, etc.), the linker further comprising an azide. In other cases, NMDA receptor ligand A comprises an alcohol group coupled to a linker containing a leaving group (e.g., Br, Cl, OMs, etc.), the linker further comprising an azide. [ka]

[0365] III Synthetic intermediate L3'-L4-R 1 Preparation of an intermediate in which L3' is a synthetic precursor of L3 in the formula. General formula L3'-L4-R 1 R 1- A linker synthesis intermediate, in which L3' is a synthesis precursor of L3, can be prepared by any of the general procedures outlined below.

[0366] In some cases, R 1 The group is coupled to the L4' linker, which is a precursor of L4, and contains an amino group, H2N-L4'-R 1 This intermediate is coupled with an L3' group containing an acidic group, where L3' is a precursor of L3. In some cases, this coupling is achieved using an activating group, such as a carbodiimide-mediated modification of a carboxylic acid (e.g., N-hydroxysuccinimide). [ka]

[0367] Preparation of the compound of formula (I) IV. The ligand-linker-azide synthesis intermediate of the general formula AY-L1-L2-N3 prepared as described above is L3'-L4-R 1 It couples with the intermediate to form the compound of formula (I). In some cases, L3' contains a triple bond. In some cases, L3' contains a cyclic alkyne. In some cases, the AZID of AY-L1-L2-N3 is L3'-L4-R 1 It reacts with the triple bond to form a triazole. [ka]

[0368] The effort of NMDA receptor ligands Ligands 1-23 were prepared according to the following procedure.

[0369] Example 1: Synthesis of Ligand 1 (1r,3R,5S,7r)-N-(12-azidododecyl)-3,5-dimethyladamantan-1-amine [ka] A mixture of memantine HCl (0.25 g, 1.2 mmol), K2CO3 (0.4 g, 2.9 mmol), and 1-azido-12-bromododecane (0.40 g, 1.4 mmol) was stirred in a sealed vial at 80°C in DMF (3 mL) for 48 hours. The reaction mixture was separated with DCM (50 mL) and water (50 mL). The organic phase was separated, and the aqueous phase was extracted with DCM (2 × 50 mL). The combined organic phase was washed with brine (20 mL), dried over Na2SO4, filtered, concentrated, and purified by silica gel column chromatography (MeOH in DCM, gradient: 0 → 10%) to obtain the title compound (144 mg, 32%) as a yellow oil.

[0370] MS(ESI), m / z 389.3[M+H] +

[0371] 1 H NMR(500MHz,DMSO-d6)d8.39(s,1H),3.31(t,J=5Hz,2H),2.71-2.74(m,2H),2.1 5(s,1H),1.40-1.60(m,10H),1.30-1.45(m,20H),1.10-1.20(m,2H),0.60(s,6H)

[0372] Example 2: Synthesis of Ligand 2 (1-Azido-15-((5S,10R)-5-methyl-10,11-dihydro-5H-5,10-epiminodibenzo[a,d][7]anulen-12-yl)-3,6,9,12-tetraoxapentadecane-15-one) [ka]

[0373] HATU (1.28 g, 3.18 mmol) was added to a solution of (+)MK-801 maleate (0.80 g, 2.37 mmol), DIPEA (2 mL, 10.83 mmol), and 1-azido-3,6,9,12-tetraoxapentadecane-15-euic acid (0.73 g, 2.51 mmol) in DCM (20 mL). The mixture was stirred for 30 minutes, then poured into a saturated NaHCO3 solution (aqueous solution, 100 mL) and extracted with DCM (300 mL x 2). The combined organic extract was dried over Na2SO4, filtered, concentrated, and purified by silica gel column chromatography (ethyl acetate in hexane, gradient: 0 → 80%) to obtain the title compound (0.49 g, 42%) as a yellow oil.

[0374] MS(ESI): m / z = 495.5[M+H] + ;517.5[M+Na] +

[0375] 1 H NMR(500MHz,CDCl3)d7.31-7.34(m,2H),7.15-7.20(m,2H),7.07-7.12(m,3H),6.89-6.90(m,1H),5.41-5. 42(m,1H),3.75-3.79(m,2H),3.56-3.66(m,15H),3.35-3.37(m,2H),2.76-2.81(m,3H),2.30-2.40(m,3H)

[0376] Example 3: Synthesis of Ligand 3 (1-Azide-N-((5R,9R,E)-11-Ethylidene-7-methyl-2-oxo-2,6,9,10-tetrahydro-5,9-methanocycloocta[b]pyridine-5(1H)-yl)-3,6,9,12-tetraoxapentadecane-15-amide) [ka] HATU (600 mg, 1.6 mmol) was added to a suspension of 1-azido-3,6,9,12-tetraoxapentadecane-15-euic acid (300 mg, 1.0 mmol), huperzine A (200 mg, 0.8 mmol), and DIPEA (1 mL) in DCM (5 mL). The mixture was stirred for 3 hours, diluted with DCM (100 mL), and washed with water (50 mL). The organic layer was separated, dried over Na2SO4, filtered, concentrated, and purified over a gradient of EtOH / siRNA (V / V, 1:3, 0 → 100%) in hexane to obtain the title compound (128 mg, 30%) as a yellow oil.

[0377] MS(ESI): m / z = 516.6[M+H] + ;538.5[M+Na] +

[0378] 1 H NMR(500MHz,CDCl3)d8.25(d,J=10Hz,1H),7.01(d,J=10Hz,1H),5.54(q,J=10Hz,1H),5.44-5.45(m,1H),3.82-3.83(m,2H),3.67-3.41( m,12H),3.10-3.41(m,2H),3.00-3.13(m,4H),2.19-2.25(m,2H),1.73(d,J=5Hz,3H),1.28(s,3H),1.30-1.40(m,1H),1.12-1.22(m,2H)

[0379] Example 4: Synthesis of Ligand 4 ((S)-2-(3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanamide)-N-benzyl-3-methoxypropanamide) [ka] (2S)-2-amino-N-benzyl-3-methoxypropanamide (168 mg, 0.81 mmol), 3-[(8-azido-3,6-dioxaocto-1-yl)oxy]propanoic acid (199 mg, 0.81 mmol), DIEA (208 mg, 1.61 mmol), and HATU (459 mg, 1.21 mmol) were added to DMF (4 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The residue was purified by reverse-phase chromatography (C18 silica gel column, mobile phase: CH3CN in water, 0% to 100% gradient over 20 minutes, detector: 254 nm UV) to obtain the title compound (170 mg, 45%) as a grayish-white solid.

[0380] MS(ESI): m / z = 460.1 [M + Na] +

[0381] 1 H NMR(CDCl3,500MHz)d7.28-7.32(m,2H),7.23-7.26(m,3H),6.90-7.01(m,2H),4.59-4.62(m,1H),4.44-4 .49(m,2H),3.89-3.92(m,1H),3.74-3.76(m,1H),3.57-3.69(m,9H),3.45-3.52(m,5H),3.30-3.40(m,5H)

[0382] Example 5: Synthesis of Ligand 5 ((S)-2-acetamide-N-(4-(2-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)benzyl)-3-methoxypropanamide) [ka]

[0383] Step 1: (S)-2-acetamide-N-(4-hydroxybenzyl)-3-methoxypropanamide To a solution of (2S)-2-amino-N-[(4-hydroxyphenyl)methyl]-3-methoxypropanamide (300 mg, 1.33 mmol) in DCM (5 mL), triethylamine (0.30 mL, 2.7 mmol) and Ac2O (68 mg, 0.67 mmol) were added dropwise at 0°C. The reaction mixture was stirred at room temperature for 2 hours, then concentrated and purified by C18 column chromatography eluting with CH3CN / H2O to obtain the title compound (100 mg, 28%) as a white semi-solid.

[0384] MS(ESI)m / z=267.1[M+H] +

[0385] Step 2: (S)-2-acetamide-N-(4-(2-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)benzyl)-3-methoxypropanamide To a solution of N-[(4S)-7-(4-hydroxyphenyl)-5-oxo-6-aza-2-oxahept-4-yl]acetamide (100 mg, 0.376 mmol) in acetonitrile (1 mL), 11-azido-3,6,9-trioxaundeca-1-ylmethanesulfonate (106 mg, 0.357 mmol) and Cs2CO3 (244 mg, 0.751 mmol) were added. The mixture was stirred at 60°C for 2 hours, then quenched by adding water (5 mL) at room temperature, and extracted with ethyl acetate (100 mL). The organic layer was washed with water (10 mL) and brine, dried over Na2SO4, filtered, concentrated, and purified by silica gel column chromatography eluting with a gradient of MeOH (0 → 5%) in DCM to obtain the title compound (84.3 mg, 47%) as a white solid.

[0386] MS(ESI): m / z = 468.2[M+H] +

[0387] 1H NMR(CDCl3,500MHz)d7.10-7.20(m,2H),6.80-6.90(m,2H),6.59-6.68(m,1H),6.38-6.42(m,1H),4.49-4.53(m,1H),4.39-4.40(m,2 H),4.10(t,J=5Hz,2H),3.85(t,J=5Hz,2H),3.78-3.82(m,1H),3.70-3.74(m,1H),3.65-3.70(m,9H),3.32-3.42(m,6H),3.03(s,3H)

[0388] Example 6: Synthesis of Ligand 6 ((S)-N-((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)-1-(3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanoyl)pyrrolidine-2-carboxamide) [ka]

[0389] Step 1: (9H-Fluoren-9-yl)methyl(S)-2-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)carbamoyl)pyrrolidine-1-carboxylate To a stirred solution of N-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-proline (2.0 g, 5.9 mmol) in DCM (20 mL), (2S,3R)-2-amino-3-hydroxybutanamide (0.70 g, 5.9 mmol) and HATU (2.7 g, 7.1 mmol) were added dropwise at room temperature. To the above mixture, DIEA (1.15 g, 8.89 mmol) was added in fractions at room temperature. The resulting mixture was stirred overnight and then extracted with ethyl acetate (3 × 250 mL). The combined organic layers were washed with saturated NaHCO3 solution (3 × 250 mL) and brine (100 mL), dried over Na2SO4, filtered, concentrated, and purified by silica gel column chromatography eluting with a gradient of MeOH (0 → 10%) in DCM to obtain the title compound (1.57 g, 60%) as a pale yellow solid.

[0390] MS(ESI)m / z=438.0[M+H] + .

[0391] Step 2: (S)-N-((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)pyrrolidine-2-carboxamide To a stirred solution of 9H-fluoren-9-ylmethyl(2S)-2-({[(1S,2R)-1-carbamoyl-2-hydroxypropyl]amino}carbonyl)tetrahydropyrrole-1-carboxylate (400 mg, 0.91 mmol) in DCM (4 mL), piperidine (0.8 mL) was added in fractions at room temperature. The resulting mixture was stirred for 2 hours. After filtration, the filtrate was concentrated under reduced pressure to obtain the crude product as a pale yellow solid, which was used directly in the next step without further purification.

[0392] Step 3: (S)-N-((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)-1-(3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanoyl)pyrrolidine-2-carboxamide To a stirred solution of (2S,3R)-3-hydroxy-2-({[(2S)-tetrahydro-1H-pyrrole-2-yl]carbonyl}amino)butanamide (220 mg, 1.02 mmol) in DCM (4 mL), 3-[(8-azido-3,6-dioxaocto-1-yl)oxy]propanoic acid (252 mg, 1.02 mmol) and HATU (583 mg, 1.53 mmol) were added dropwise at room temperature. DIEA (396 mg, 3.07 mmol) was added in fractions to the mixture. The resulting mixture was stirred for a further 1 hour. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by reverse-phase chromatography (C18 silica gel column, mobile phase: ACN in water, 10% to 90% gradient over 20 minutes, detector: 254 nm UV) to obtain the title compound (247 mg, 51%) as a yellow oil.

[0393] MS(ESI): m / z = 445.4[M+H] + ,467.4[M+Na] +

[0394] 1 H NMR(CDCl3,500MHz)d4.50-4.55(m,2H),4.40-4.45(m,1H),3.86-3.91(m,2H),3.50-3.60(m,12H),3.30-3.40(m,2H) ),3.25(s,2H,br),2.80-2.90(m,1H),2.48-2.51(m,1H),2.20-2.30(m,2H),2.00-2.10(m,2H),1.16(d,J=5.0Hz,3H)

[0395] Example 7: Ligand 7 (2-((2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)amino)-2-(2-chlorophenyl)cyclohexane-1-one (second eluate) and Ligand 8 Synthesis of N-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-2-(2-chlorophenyl)cyclopentane-1-carboxamide (first eluate) [ka]

[0396] Step 1: 1-(2-chlorophenyl)-2-oxocyclohexylmethanesulfonate To a stirred solution of 2-(2-chlorophenyl)-2-hydroxycyclohexane-1-one (0.80 g, 3.5 mmol, prepared as described in J.Org.Chem.2020, 85(13), 8656-8664) in THF (0.4 mL), triethylamine (1.7 mL, 12 mmol) and methanesulfonyl chloride (1.22 g, 10.7 mmol) were added in fractions at 0°C. The resulting mixture was stirred for 1 hour at 0°C under a nitrogen atmosphere. The reaction was quenched by adding water (25 mL) at room temperature and extracted with ethyl acetate (2 × 250 mL). The combined organic layer was washed with water (2 × 25 mL) and brine (25 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was used directly in the next step without further purification. MS(ESI): m / z = 301.0 [MH] - .

[0397] Step 2: 2-((2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)amino)-2-(2-chlorophenyl)cyclohexane-1-one and N-(2-(2-(2-(azidomethoxy)ethoxy)ethoxy)ethyl)-2-(2-chlorophenyl)cyclopentan-1-carboxamide To a stirred solution of 1-(2-chlorophenyl)-2-oxocyclohexylmethanesulfonate (0.80 g, 2.64 mmol) in THF (1 mL), triethylamine (1.84 mL, 13.2 mmol) was added dropwise at 0°C. 2-[(8-azido-3,6-dioxaoccto-1-yl)oxy]ethane-1-amine (1.73 g, 7.93 mmol) was added dropwise at 0°C, and the resulting mixture was stirred overnight at room temperature. The reaction was quenched by adding water (10 mL), and the mixture was extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with water (100 mL) and brine (100 mL), dried over Na2SO4, filtered, concentrated, and purified by reverse-phase flash chromatography (C18 silica gel column, mobile phase: ACN in water, 10% → 90% gradient over 20 minutes, detector: 254 nm UV) to obtain the following.

[0398] Compound of the first eluate N-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-2-(2-chlorophenyl)cyclopentan-1-carboxamide (ligand 8, 181.9 mg, 15%) as a pale yellow solid.

[0399] MS(ESI): m / z = 425.2[M+H] +

[0400] 1 H NMR(DMSO-d6,400MHz)d7.80-7.90(m,1H),7.15-7.45(m,4H),3.20-3.70(m,15H),3.05-3.15(m,2 H),2.80-2.95(m,1H),2.10-2.20(m,1H),1.90-2.05(m,1H),1.70-1.80(m,3H),1.35-1.50(m,1H)

[0401] Compounds of the second eluate : 2-[(11-azido-3,6,9-trioxaundecay-1-yl)amino]-2-(2-chlorophenyl)cyclohexane-1-one (ligand 7, 199.9 mg, 16%) as a pale yellow solid

[0402] MS(ESI): m / z = 425.2[M+H] +

[0403] 1 H NMR(DMSO-d6,400MHz)d7.65-7.70(m,1H),7.25-7.40(m,3H),3.42-3.62(m,11H),3.34-3.42(m,2H),2.65- 2.75(m,1H),2.50-2.60(m,1H),2.25-2.45(m,3H),2.05-2.15(m,1H),1.80-1.90(m,3H),1.60-1.75(m,2H)

[0404] Example 8: Synthesis of Ligand 9 (1-Azido-N-((1r,3R,5S,7r)-3,5-Dimethyladamantan-1-yl)-3,6,9,12-Tetraoxapentadecane-15-amide) [ka] A suspension of azide-PEG4-NHS ester (541 mg, 1.395 mmol), memantine hydrochloride (300 mg, 1.395 mmol), and DIPEA (0.43 ml, 3.1 mmol) in THF (10 mL) was stirred at 60°C for 3 hours. The reaction mixture was concentrated, and the residue was purified by flash chromatography using hexane and ethyl acetate (0 → 100%) on a 10 g, 20 micrometer Biotage column to obtain the title compound (293 mg, 46%) as a beige solid.

[0405] MS(ESI): m / z = 453[M+H] +

[0406] 1 H NMR(499MHz,DMSO-d6)δ7.29(s,1H),3.63-3.56(m,2H),3.56-3.52(m,6H),3.52-3.43(m,8H),3.39(dd,J=5.6,4.3Hz,2H),2.24(t,J=6.6Hz,2H) ,2.05(p,J=3.1Hz,1H),1.73(d,J=3.2Hz,2H),1.60-1.50(m,4H),1.30( dd,J=12.3,3.1Hz,2H),1.23(d,J=12.1Hz,2H),1.09(s,2H),0.80(s,6H)

[0407] Example 9: Synthesis of Ligand 10 (1-Azido-15-((5R,10S)-5-methyl-10,11-dihydro-5H-5,10-epiminodibenzo[a,d][7]anulen-12-yl)-3,6,9,12-tetraoxapentadecane-15-one) [ka] DIPEA (3.0 mL, 17 mmol) was added to a suspension of (-)MK801 maleate (0.86 g, 2.31 mmol) in DCM (20 mL) to obtain a clear solution. N3-PEG4-COOH (0.9 g, 3.09 mmol) and HATU (1.2 g, 3.16 mmol) were added simultaneously. The initial suspension turned into a yellow solution. The reaction mixture was stirred for 1 hour, then poured into saturated NaHCO3 aqueous solution (200 mL) and diluted with DCM (500 mL). The organic layer was separated, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (80 g silica gel column, eluted with 30% EtOH in toluene, gradient: 0 → 80% in hexane) to obtain the title compound (836 mg, 58%) as a pale yellow oil.

[0408] MS(ESI): m / z = 495.3[M+H] + ,517.5[M+Na] +

[0409] 1 H NMR(499MHz,CDCl3)δ7.37-7.29(m,2H),7.21-7.14(m,2H),7.13-7.03(m,3H),6.93-6.84(m,1H),5. 43(s,1H),3.85-3.72(m,2H),3.70-3.52(m,16H),3.40-3.33(m,2H),2.89-2.70(m,2H),2.37(s,3H)

[0410] Example 10: Synthesis of Ligand 11 (1-Azido-15-(3-Chloro-5-methyl-10,11-dihydro-5H-5,10-epiminodibenzo[a,d][7]anulen-12-yl)-3,6,9,12-tetraoxapentadecane-15-one) [ka] Using 3-chloro-5-methyl-10,11-dihydro-5H-5,10-epiminodibenzo[a,d][7]annulene as a starting material (prepared as described in J.Med.Chem. 1996, 39, 5257-5266), compound 13 was prepared according to the procedure described for compound 12 to obtain the title compound as a yellow oil.

[0411] MS(ESI): m / z = 529.2[M+H] + ,551.2[M+Na] +

[0412] 1 H NMR(400MHz,DMSO-d6)δ7.48-7.32(m,2H),7.26-7.14(m,4H),6.96(d,J=8.2Hz,1H),5.6 5(d,J=5.4Hz,1H),3.66-3.43(m,17H),3.39-3.36(m,2H),2.79-2.61(m,3H),2.24(s,3H)

[0413] Example 11: Synthesis of Ligand 12 ((4aR,5R,10bR)-1-(1-azido-3,6,9,12-tetraoxapentadecane-15-oil)-12-methyl-2,3,4,4a,5,6-hexahydro-1H-5,10b-propaeno-1,7-phenanthroline-8(7H)-one) [ka] Huperzine B (486 mg, 1.90 mmol) was dissolved in DCM (20 mL), and DIPEA (1 mL) was added. Azido-PEG-4 acid (694 mg, 2.38 mmol) and HATU (786 mg, 2.07 mmol) were added, and the solution was stirred for 2 hours. Subsequently, the reaction mixture was poured into saturated sodium bicarbonate solution (100 mL) and diluted with DCM (500 mL). The organic layer was separated, dried over anhydrous sodium sulfate, filtered, concentrated, and purified using a silica gel RediSepGold column (80 g, eluted with 0 → 80% hexane and Green's solvent) to obtain the title compound (466 mg, 48%) as a pale yellow oil.

[0414] MS(ESI): m / z = 530.2[M+H] + ,552.6[M+Na] +

[0415] 1 H NMR(499MHz,CDCl3)δ8.07(d,J=8.4Hz,1H),6.99(d,J=8.3Hz,1H),5.50(d,J=5.6Hz,1H),3.86(t,J =6.3Hz,2H),3.68-3.62(m,13H),3.60(t,J=5.1Hz,2H),3.37-3.23(m,2H),3.02(dd,J=18.1,5.7Hz ,1H),2.90(t,J=6.3Hz,2H),2.85-2.75(m,1H),2.70-2.56(m,1H),2.51-2.38(m,1H),2.34-2.14(m ,2H),1.90(d,J=16.9Hz,1H),1.84-1.77(m,1H),1.67-1.57(m,2H),1.57-1.51(m,1H),1.51(s,3H)

[0416] Example 12: Synthesis of Ligand 13 ((S)-1-((2S,4R)-1-(L-Threonyl)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-2-carbonyl)-N-((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)pyrrolidine-2-carboxamide) [ka]

[0417] Step 1: (2S,4R)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid 1-tert-butyl 2-methyl(2S,4R)-4-hydroxypyrrolidine-1,2-dicarboxylate (2.0 g, 8.2 mmol) in DMF (20 mL) was mixed with NaH (0.65 g, 16.3 mmol, 60% in oil) in fractional amounts at 0°C. The resulting mixture was stirred for 30 minutes at 0°C under a nitrogen atmosphere. 11-azido-1-iodo-3,6,9-trioxaundecane (2.68 g, 8.15 mmol) was added dropwise at 0°C, and the resulting mixture was stirred at room temperature for 2 hours. The reaction was quenched by adding ice water, extracted with ethyl acetate (500 mL), and the layer was separated. The organic layer was concentrated to obtain methyl ester (3.4 g, 10% purity) as a yellow oil.

[0418] MS(ESI)m / z=447.3[M+H] + .

[0419] The aqueous layer was acidified with HCl aqueous solution (1N), extracted with ethyl acetate (500 mL), the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound containing DMF (4.6 g, crude product), which was used in the next step without further purification.

[0420] MS (ESI, negative mode) m / z = 431.0 [MH] - .

[0421] Step 2: tert-butyl(2S,4R)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)-2-((S)-2-((benzyloxy)carbonyl)pyrrolidine-1-carbonyl)pyrrolidine-1-carboxylate To a stirred solution of (2S,4R)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid (4.6 g, crude product obtained from step 1) and benzyl L-proline hydrochloride (1.54 g, 6.38 mmol) in DCM (46 mL), HATU (2.43 g, 6.38 mmol) was added in portions at room temperature. DIEA (3.30 g, 25.5 mmol) was added dropwise at room temperature, and the resulting mixture was stirred overnight at room temperature. The reaction product was quenched by adding water (10 mL) and diluted with ethyl acetate (100 mL). The combined organic layers were washed with water and brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by reverse-phase flash chromatography (C18 silica gel column, mobile phase: ACN in water, 10% to 90% gradient over 20 minutes, detector: 254 nm UV) to obtain the title compound (540 mg, purity 50%, 26.6%) as a brown oil. MS(ESI): m / z = 620.5[M+H] +

[0422] Step 3: ((2S,4R)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)-1-(tert-butoxycarbonyl)pyrrolidine-2-carbonyl)-L-proline To a stirred solution of tert-butyl(2S,4R)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)-2-((S)-2-((benzyloxy)carbonyl)pyrrolidine-1-carbonyl)pyrrolidine-1-carboxylate (500 mg, 0.807 mmol) in THF (5 mL), NaOH (1 N, 4.03 mL, 4.03 mmol) was added dropwise at room temperature. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere, and then acidified (pH 2) with HCl solution (1 N). The resulting mixture was extracted with ethyl acetate, the combined organic layers were washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound (500 mg, crude) as a yellow oil, which was used directly in the next step without further purification.

[0423] MS(ESI)m / z=530.4[M+H] +

[0424] Step 4: tert-butyl(2S,4R)-2-((S)-2-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)carbamoyl)pyrrolidine-1-carbonyl)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-1-carboxylate To a stirred solution of ((2S,4R)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)-1-(tert-butoxycarbonyl)pyrrolidine-2-carbonyl)-L-proline (500 mg, 0.944 mmol) in DCM (5 mL), (2S,3R)-2-amino-3-hydroxybutanamide hydrochloride (291.91 mg, 1.888 mmol) was added in fractions at room temperature. DIEA (366 mg, 2.83 mmol) was added dropwise at room temperature, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated and purified by reverse-phase flash chromatography (C18 silica gel column, mobile phase: ACN in water, 10% → 90% gradient over 20 minutes, detector: 254 nm UV) to obtain the title compound (550 mg, 93%) as a pale yellow oil.

[0425] MS(ESI)m / z=630.5[M+H] +

[0426] Step 5: (S)-N-((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)-1-((2S,4R)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-2-carbonyl)pyrrolidine-2-carboxamide To a solution of tert-butyl(2S,4R)-2-((S)-2-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)carbamoyl)pyrrolidine-1-carbonyl)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-1-carboxylate (550 mg, 0.873 mmol) in DCM (5 mL), TFA (5 mL, 65 mmol) was added dropwise at room temperature. The resulting mixture was stirred for 1 hour at room temperature under a nitrogen atmosphere and concentrated to obtain the title compound (1.00 g, crude) as a yellow oil, which was used directly in the next step without further purification. MS(ESI)m / z = 530.3[M+H] +

[0427] Step 6: (9H-Fluoren-9-yl)methyl((2S,3R)-1-((2S,4R)-2-((S)-2-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)carbamoyl)pyrrolidine-1-carbonyl)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-1-yl)-3-hydroxy-1-oxobutan-2-yl)carbamate To a stirred solution of (S)-N-((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)-1-((2S,4R)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-2-carbonyl)pyrrolidine-2-carboxamide (500 mg, 0.944 mmol) in DCM (5 mL), N-{[(9H-fluoren-9-ylmethyl)oxy]carbonyl}-L-threonine (322 mg, 0.944 mmol) was added in fractions at room temperature. HATU (538 mg, 1.42 mmol) and DIEA (732 mg, 5.67 mmol) were added in fractions, and the resulting mixture was stirred overnight at room temperature. The resulting solution was concentrated and purified by C18 chromatography (ACN / H2O) to obtain the title compound (300 mg, 37%) as a yellow solid.

[0428] MS(ESI)m / z=853.4[M+H] +

[0429] Step 7: (S)-1-((2S,4R)-1-(L-Threonyl)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-2-carbonyl)-N-((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)pyrrolidine-2-carboxamide To a stirred solution of (9H-fluoren-9-yl)methyl((2S,3R)-1-((2S,4R)-2-((S)-2-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)carbamoyl)pyrrolidine-1-carbonyl)-4-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)pyrrolidine-1-yl)-3-hydroxy-1-oxobutan-2-yl)carbamate (260 mg, 0.305 mmol) in DCM (3 mL), Et2NH (0.8 mL) was added dropwise at room temperature. The resulting mixture was stirred for 2 hours at room temperature under a nitrogen atmosphere, then concentrated and extracted with water. The aqueous layer was washed with ethyl acetate, and the combined aqueous layer was freeze-dried overnight. Then, it was evaporated simultaneously with ACN and freeze-dried again to obtain the title compound (150.6 mg, 73.8%) as a pale yellow solid.

[0430] MS(ESI)m / z=631.5[M+H] +

[0431] 1H NMR(499MHz,DMSO-d6)δ7.42(d,J=8.4Hz,1H),7.10-7.04(m,2H),4.86(s,1H),4.58(t,J=8.0Hz,1H) ,4.43-4.34(m,1H),4.22-4.15(m,1H),4.10-3.98(m,2H),3.85-3.76(m,1H),3.75-3.65(m,1H),3.6 7-3.61(m,1H),3.64-3.57(m,3H),3.60-3.48(m,12H),3.50-3.42(m,1H),3.42-3.36(m,2H),3.29-3 .23(m,1H),2.34-2.26(m,1H),2.07-1.97(m,1H),1.97-1.87(m,3H),1.90(s,1H),1.15-0.97(m,6H)

[0432] Example 13: Ligand 14 (N-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-2-(1-((1R,2S)-1-hydroxy-1-(4-hydroxyphenyl)propan-2-yl)piperidine-4-yl)acetamide and Ligand 15 Synthesis of N-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-2-(1-((1R,2R)-1-hydroxy-1-(4-hydroxyphenyl)propan-2-yl)piperidine-4-yl)acetamide) [ka]

[0433] Step 1: Ethyl 2-(1-(1-(4-((tert-butyldimethylsilyl)oxy)phenyl)-1-oxopropan-2-yl)piperidine-4-yl)acetate To a stirred solution of 2-bromo-1-(4-((tert-butyldimethylsilyl)oxy)phenyl)propan-1-one (1.2 g, 3.5 mmol, prepared as described in Bioorg. Med. Chem. Lett. 2007, 5558) in ethanol (20 mL), TEA (0.53 g, 5.24 mmol) and ethyl xahydropyridine-4-yl acetate (0.90 g, 5.2 mmol) were added in fractions at room temperature. The resulting mixture was heated at 80 °C under nitrogen for 3 hours, and then quenched with water (5 mL) at room temperature. The mixture was extracted with DCM (200 mL), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography eluting with PE:EA (2:1 (v / v)) to obtain the title compound (1.0 g, 66%) as a colorless oil.

[0434] MS(ESI)m / z=434.2[M+H] +

[0435] 1 H NMR(300MHz,CDCl3)δ8.02(d,2H),6.85(d,J=8.3Hz,2H),4.18-4.05(m,2H),4.02(m,1H),2.89(m,1 H),2.79(m,1H),2.39(m,1H),2.20(m,3H),1.68(m,5H),1.31-1.19(m,6H),0.99(s,9H),0.24(s,6H)

[0436] Step 2: Ethyl 2-(1-(1-(4-((tert-butyldimethylsilyl)oxy)phenyl)-1-hydroxypropan-2-yl)piperidine-4-yl)acetate To a solution of ethyl 2-(1-(1-(4-((tert-butyldimethylsilyl)oxy)phenyl)-1-oxopropan-2-yl)piperidine-4-yl)acetate (900 mg, 2.075 mmol) in methanol (10 mL), NaBH4 (157 mg, 4.15 mmol) was added in batch mode at 0°C. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere, then quenched by adding water and extracted with ethyl acetate (200 mL). The combined organic layers were washed with water and brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound (1.0 g, crude) as a yellow oil, which was used directly without further purification.

[0437] MS(ESI)m / z=436.3[M+H] +

[0438] Step 3: 2-(1-(1-hydroxy-1-(4-hydroxyphenyl)propan-2-yl)piperidine-4-yl)acetic acid To a stirred solution of ethyl 2-(1-(1-(4-((tert-butyldimethylsilyl)oxy)phenyl)-1-hydroxypropan-2-yl)piperidine-4-yl) acetate (1.0 g, 2.3 mmol) in THF (10 mL), aqueous NaOH solution (1 N, 13.8 mL, 13.8 mmol) was added dropwise at 25°C. The resulting mixture was stirred overnight at room temperature and then quenched by dropwise addition of acetic acid (0.92 mL, 16 mmol) at room temperature. The mixture was concentrated and purified by reverse-phase column chromatography (Combi-Flash® C18 column, ACN / H2O) to obtain a trans / cis mixture of the title compound (450 mg, 67%) as a white solid.

[0439] MS(ESI): m / z = 294.2[M+H] +

[0440] 1¹H NMR (300MHz, DMSO-d6) δ 7.14-7.02 (m, 2H), 6.73-6.61 (m, 2H), 4.57 (d, J=4.8Hz, 1H, corresponding to cis isomer), 4.12 (d, J=9.4Hz, 1H, corresponding to trans isomer), 2.80 (t, J=12.7Hz, 1H), 2.65 (d, J=13.5Hz, 1H), 2.34 (d, NMR spectrum showing a mixture of cis isomers and trans isomers (5:7): J=11.5Hz, 0H), 2.10(m,J=24.1,6.4Hz,2H), 1.70(d,J=12.4Hz,2H), 1.56(d,J=11.5Hz,1H), 1.36-1.25(m,0H), 0.88(d,J=6.7Hz,1H), 0.63(d,J=6.6Hz,2H)

[0441] Step 4: N-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-2-(1-((1R,2S)-1-hydroxy-1-(4-hydroxyphenyl)propan-2-yl)piperidine-4-yl)acetamide (compound 14) and N-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-2-(1-((1R,2R)-1-hydroxy-1-(4-hydroxyphenyl)propan-2-yl)piperidine-4-yl)acetamide (compound 15) To a solution of 2-(1-(1-hydroxy-1-(4-hydroxyphenyl)propan-2-yl)piperidine-4-yl)acetic acid (270 mg, 0.920 mmol) in DMF (3 mL), HOBt (187 mg, 1.38 mmol), EDCI (265 mg, 1.38 mmol), and DIEA (1070 mg, 8.283 mmol) were added, and the mixture was stirred for 5 minutes at room temperature under a nitrogen atmosphere. 2-[(8-azido-3,6-dioxaocto-1-yl)oxy]ethane-1-amine (301 mg, 1.38 mmol) was added dropwise at room temperature, and the resulting mixture was stirred at 30°C for 36 hours. The mixture was concentrated and purified by reverse-phase flash chromatography (C18 silica gel column, mobile phase: ACN in water, 10% → 90% gradient over 20 minutes, detector: 254 nm UV), and further purified by preparative HPLC (column: XBridge Prep OBD C18 Column (19 × 150 mm, 5 m), mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN, flow rate: 25 mL / min, gradient: isocratic (18 → 28), wavelength: 254 nm / 210 nm) to obtain the following.

[0442] Ligand 14 (Rt=4.9 min, 80.3 mg, 17%, racemic (cis)), as a yellow oil

[0443] MS(ESI): m / z = 494.4[M+H] +

[0444] 1H NMR(499MHz, DMSO-d6)δ9.13(s,1H),7.80(t,J=5.7Hz,1H),7.09-7.00(m,2H),6.68-6.59(m,2H),4.79(d,J=4. 5Hz,1H),4.52(t,J=4.7Hz,1H),3.60(dd,J=5.6,4.3Hz,2H),3.57-3.47(m,8H),3.41-3.35(m,4H),3.16(q,J=5 .8Hz,2H),2.77(d,J=11.0Hz,1H),2.65(d,J=11.3Hz,1H),2.53(t,J=6.7Hz,1H),2.28(td,J=11.5,2.3Hz,1H), 2.12(td,J=11.5,2.3Hz,1H),1.92(d,J=7.0Hz,2H),1.40-1.60(m,3H),0.90-1.00(m,2H),0.88(d,J=6.7Hz,3H)

[0445] Ligand 15 :(Rt=7.3 points, 119.6mg, 25%, ラセミ(トランス)), yellow oil としてのもの

[0446] MS(ESI): m / z=494.45[M+H] +

[0447] 1 H NMR(499MHz,DMSO-d6)δ9.27(s,1H),7.86(t,J=5.7Hz,1H),7.13-7.07(m,2H),6.72-6.66(m,2H),4.81 (s,1H),4.11(d,J=9.4Hz,1H),3.63-3.55(m,2H),3.59-3.47(m,8H),3.43-3.36(m,3H),3.31(s,1H),3. 19(q,J=5.8Hz,2H),2.76(d,J=11.3Hz,1H),2.63(d,J=11.5Hz,1H),2.51-2.41(m,2H),2.11-2.03(m,1H ),2.01(d,J=6.7Hz,2H),1.69-1.61(m,3H),1.32-1.21(m,1H),1.18-1.07(m,1H),0.63(d,J=6.6Hz,3H)

[0448] Example 14: Synthesis of リガンド16 1-Azido-15-(3-chloro-5-methyl-10,11-dihydro-5H-5,10-epiminodibenzo[a,d][7]-anulen-12-yl)-3,6,9,12-tetraoxapentadecane-15-one [ka]

[0449] DIPEA (1.0 mL, 5.74 mmol) was added to a suspension of 3-chloro-5-methyl-10,11-dihydro-5H-5,10-epiminodibenzo[a,d][7]anulene (prepared as described in J.Med.Chem. 1990, 33, 789-808, 90 mg, 0.35 mmol) in DCM (2 mL). Subsequently, HATU (201 mg, 0.53 mmol) and N3PEG4COOH (0.9 g, 3.09 mmol) were added all at once, and the mixture was stirred for 1 hour. The mixture was poured into saturated NaHCO3 aqueous solution (20 mL) and diluted with DCM (50 mL). The organic layer was separated, dried over Na2SO4, filtered, concentrated, and purified by column chromatography (elution using a 25g silica gel column with 30% EtOH in toluene and a 0-80% gradient in hexane) to obtain the title compound (90mg, 49%) as a pale yellow oil.

[0450] MS(ESI)m / z=529.4[M+H] + ,551.4[M+Na] +

[0451] 1 H NMR(499MHz,CDCl3)δ7.36-7.28(m,2H),7.24-7.16(m,2H),7.11-6.99(m,2H),6.83(d,J=8.1Hz,1H),5.43(s ,1H),3.79(dt,J=9.5,6.9Hz,2H),3.71-3.46(m,15H),3.37(t,JJ=5.0Hz,2H),2.90-2.62(m,3H),2.35(s,3H)

[0452] Example 15: Synthesis of Ligand 17 (5S,10R)-12-(12-azidododecyl)-5-methyl-10,11-dihydro-5H-5,10-epiminodibenzo-[a,d][7]annulene [ka] K2CO3 (1.3 g, 9.4 mmol) was added to (+)-MK801 maleate (0.51 g, 1.5 mmol) and 1-azido-12-bromododecane (0.6 g, 1.37 mmol) in DMF (5 mL), and the mixture was heated to 80°C. The second portion of 1-azido-12-bromododecane (0.3 g, 0.69 mmol) was added, and the mixture was heated to 90°C for 5 hours. After cooling, the mixture was extracted with ethyl acetate (300 mL), washed with lithium chloride solution (aqueous solution, 10%) and brine, dried over Na2SO4, filtered, concentrated, and purified by flash column chromatography to obtain the title compound (340 mg, 53%) as a yellow oil.

[0453] MS(ESI): m / z = 431.4[M+H] +

[0454] 1 H NMR(499MHz,CDCl3)δ7.31-7.28(m,1H),7.24-7.20(m,1H),7.13-7.01(m,5H),6.93-6.87(m,1H),4.56(d,J=5.2Hz,1H),3.3 7-3.19(m,3H),2.63-2.52(m,1H),2.50-2.45(m,1H),2.40-2.29(m,1H),1.86(s,3H),1.73-1.54(m,4H),1.41-1.18(m,16H)

[0455] Example 16: Synthesis of Ligand 18 (S)-N-(((1R,3S,5S,7S)-adamantan-2-yl)methyl)-2-(3-(2-(2-(2-azidoethoxy)ethoxy)-ethoxy)propanamide)-3-methoxypropanamide [ka] To a stirred solution of (2S)-2-[(1-azido-12-oxo-3,6,9-trioxadodeca-12-yl)amino]-3-methoxypropanoic acid (200 mg, 0.574 mmol) in DMF (3 mL), adamantane-2-ylmethaneamine (95 mg, 0.57 mmol), HOBt (116 mg, 0.861 mmol), and EDCI (165 mg, 0.861 mmol) were added in batches at room temperature. DIEA (371 mg, 2.87 mmol) was added dropwise, and the mixture was stirred overnight at room temperature under a nitrogen atmosphere. The reaction was quenched with water and extracted with ethyl acetate. The combined organic extracts were washed with saturated NaHCO3 aqueous solution and brine, dried over Na2SO4, filtered, concentrated, and purified by reverse-phase flash chromatography (SunFire C18 OBD Prep Column (19×150mm, 5m), mobile phase A: water (0.1% FA), mobile phase B: CH3CN, flow rate: 25 mL / min, gradient: 52% B → 62% B over 8 minutes, wavelength: 254 nm / 220 nm, RT1 (min) 4.37) to obtain the title compound (123 mg, 42.8%) as a yellow oil.

[0456] MS(ESI)m / z=496.3[M+H] +

[0457] 1 H NMR(400MHz,CDCl3)δ7.01(d,J=7.2Hz,1H),6.51(s,1H),4.56-4.47(m,1H),3.87-3.68(m,3H),3.72- 3.59(m,10H),3.46-3.30(m,8H),2.62-2.43(m,2H),1.94-1.79(m,7H),1.69(s,6H),1.60-1.50(m,2H)

[0458] Example 17: Synthesis of Ligand 19 (S)-2-(3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanamide)-N-(4-fluorobenzyl)-3-methoxypropanamide [ka]

[0459] Step 1: (2S)-2-amino-3-methoxypropanoate hydrochloride To a stirred solution of (2S)-2-amino-3-methoxypropano hydrochloride (5.0 g, 32 mmol) in methanol (50 mL), thionyl chloride (7.65 g, 64.3 mmol) was added dropwise at 0°C. The resulting mixture was stirred for 2 hours at 60°C under a nitrogen atmosphere. After cooling the mixture to room temperature, it was concentrated to obtain the title compound (6.0 g, 100%) as a grayish-white solid, which was used directly without further purification.

[0460] MS(ESI)m / z=134.1[M+H] +

[0461] 1 H NMR(300MHz,DMSO-d6)δ8.83(s,3H),4.70(s,2H),4.27(s,1H),3.81(d,J=3.5Hz,2H),3.75(s,3H),3.29(s,3H)

[0462] Step 2: (2S)-2-[(1-azido-12-oxo-3,6,9-trioxadodeca-12-yl)amino]-3-methoxypropanoate To a stirred solution of methyl(2S)-2-amino-3-methoxypropanoate hydrochloride (1.17 g, 6.88 mmol) in DMF (17 mL), 3-[(8-azido-3,6-dioxaocto-1-yl)oxy]propanoic acid (1.70 g, 6.88 mmol), HOBt (1.39 g, 10.3 mmol), and EDCI (1.98 g, 10.3 mmol) were added in batches at room temperature. Subsequently, DIEA (2.67 g, 20.627 mmol) was added dropwise, and the mixture was stirred overnight under a nitrogen atmosphere. The reaction was quenched by adding water, and then extracted with ethyl acetate. The combined organic extracts were washed with water, saturated NaHCO3 solution, and brine, dried over Na2SO4, filtered, and concentrated to obtain the title compound (1.6 g, 64%) as a yellow oil, which was used directly in the next step without further purification.

[0463] MS(ESI)m / z=349.2[M+H] +

[0464] 1 H NMR(400MHz,CDCl3)δ7.04(d,J=8.2Hz,1H),4.78-4.70(m,1H),3.86-3.73(m,3H),3.76(s,3H) ,3.77-3.61(m,11H),3.64-3.57(m,1H),3.39(t,J=5.1Hz,2H),3.34(s,3H),2.66-2.50(m,2H)

[0465] Step 3: N-(3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanoyl)-O-methyl-L-serine To a stirred solution of methyl(2S)-2-[(1-azido-12-oxo-3,6,9-trioxadodeca-12-yl)amino]-3-methoxypropanoate (1.6 g, 4.4 mmol) in THF (16 mL), sodium hydroxide solution (17.7 mL, 17.7 mmol, 1 mol / L) was added dropwise at room temperature, and the resulting mixture was stirred for 2 hours. Subsequently, the mixture was acidified to pH 3 using Dowex resin (Beijing Innochem Science & Technology Co. Ltd.), washed with acetonitrile, filtered, and the filtrate was concentrated to obtain the title compound (1.2 g, 78%) as a yellow oil, which was used directly in the next step without further purification.

[0466] MS(ESI)m / z=349.2[M+H] +

[0467] 1 H NMR(300MHz,CDCl3)δ7.23(d,J=7.6Hz,1H),5.37(s,3H),4.76-4.65(m,1H),3.91-3.81(m,1 H),3.82-3.70(m,3H),3.70-3.55(m,13H),3.44-3.39(m,2H),3.38(s,3H),2.69-2.40(m,2H)

[0468] Step 4: (S)-2-(3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanamide)-N-(4-fluorobenzyl)-3-methoxypropanamide To a stirred solution of N-(3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanoyl)-O-methyl-L-serine (200 mg, 0.574 mmol) in DMF (3 mL), (4-fluorophenyl)methanamine (71 mg, 0.57 mmol), HOBt (116 mg, 0.861 mmol), and EDCI (165 mg, 0.861 mmol) were added in batches at room temperature. Subsequently, DIEA (371 mg, 2.87 mmol) was added dropwise, and the mixture was stirred under a nitrogen atmosphere for 3 hours. The reaction was quenched with water and extracted with ethyl acetate. The combined organic extracts were washed with saturated NaHCO3 solution and brine, dried over Na2SO, filtered, concentrated, and purified by reverse-phase flash chromatography (SunFire C18 OBD Prep Column (19×150mm, 5m), mobile phase A: water (0.1% FA), mobile phase B: CH3CN, flow rate: 25 mL / min, gradient: 8 min from 33% B to 43% B, wavelength: 254 nm / 220 nm, Rt(min): 5.18) to obtain the title compound (99.5 mg, 38%) as a white solid.

[0469] MS(ESI)m / z=456.1[M+H] +

[0470] 1 H NMR(400MHz,CDCl3)δ7.27-7.18(m,2H),7.05-6.95(m,3H),4.64-4.56(m,1H),4.52-4.43(m,1H),4.43-4.35(m, 1H),3.96-3.88(m,1H),3.82-3.72(m,2H),3.71-3.53(m,6H),3.57-3.43(m,5H),3.38(s,5H),2.61-2.38(m,2H)

[0471] Example 18: Synthesis of Ligand 20 (S)-2-(3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanamide)-3-methoxy-N-(4-methylbenzyl)propanamide [ka] (S)-2-(3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanamide)-N-(4-fluorobenzyl)-3-methoxypropanamide was prepared using p-tolylmethaneamine instead of (4-fluorophenyl)methaneamine, and the title compound was obtained as a white solid (95% purity).

[0472] MS(ESI)m / z = 452.10[M+H] + ,474.10[M+Na] +

[0473] Example 19: Synthesis of Ligand 21 N-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-1-(2-chlorophenyl)pyrrolidine-2-carboxamide [ka]

[0474] Step 1: (2-chlorophenyl)proline To a stirred solution of 1-chloro-2-iodobenzene (1.0 g, 4.194 mmol) in isopropanol (10 mL), DL-proline (0.58 g, 5.033 mmol), copper(I) iodide (0.16 g, 0.839 mmol), tripotassium phosphate (1.78 g, 8.388 mmol), and ethylene glycol (0.468 mL, 8.388 mmol) were added at room temperature. The resulting mixture was stirred for 20 hours at 85°C under a nitrogen atmosphere. The reaction was quenched with water, the pH was adjusted to 3 with HCl (6 M), extracted with ethyl acetate, washed with brine, dried over sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (MeOH / DCM) to obtain the title compound (200 mg, 21%) as a yellow oil.

[0475] MS(ESI)m / z=226.1,[M+H] +

[0476] Step 2: N-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-1-(2-chlorophenyl)pyrrolidine-2-carboxamide To a stirred solution of (2-chlorophenyl)proline (0.22 g, 0.975 mmol) in N,N-dimethylmethaneamide (2.2 mL), HOBT (0.20 g, 1.462 mmol), EDCI (0.28 g, 1.462 mmol), 2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethane-1-amine (0.21 g, 0.975 mmol) and DIEA (0.50 g, 3.900 mmol) were added at room temperature. The resulting mixture was stirred for 2 hours at room temperature under a nitrogen atmosphere. The reaction was quenched with water, extracted with ethyl acetate, washed with brine, dried over sodium sulfate, filtered, concentrated, and purified by preparative HPLC to obtain the title compound (234.2 mg, 54%) as a yellow oil.

[0477] MS(ESI)m / z = 426.3[M+H] +

[0478] 1H NMR (300MHz, chloroform-d) δ7.40-7.27(m,2H),7.24-7.12(m,1H),7.12-7.02(m,1H),7.01-6.90(m,1H),4.32-4.21(m,1H),4.01-3.88(m,1H),3.71- 3.59(m,6H),3.59-3.52(m,2H),3.53-3.44(m,2H),3.44-3.34(m,4H),3. 35-3.25(m,2H),3.06-2.92(m,1H),2.51-2.34(m,1H),2.21-1.62(m,4H)

[0479] Example 20: Synthesis of Ligand 22 3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)-1-(2-(2-chlorophenyl)pyrrolidine-1-yl)propan-1-one [ka] To a stirred solution of 3-[(8-azido-3,6-dioxaocto-1-yl)oxy]propanoic acid (300 mg, 1.213 mmol) in DMF (3 mL), 2-(2-chlorophenyl)tetrahydropyrrole (220.42 mg, 1.213 mmol), HOBt (245.94 mg, 1.820 mmol), and EDCI (348.90 mg, 1.820 mmol) were added in batches at room temperature. DIEA (784.13 mg, 6.067 mmol) was added dropwise, and the resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The reaction was quenched by adding saturated NaHCO3 aqueous solution, and then extracted with ethyl acetate. The combined organic extracts were washed with saturated NaHCO3 solution and brine, dried over Na2SO4, filtered, concentrated, and purified by reverse-phase flash chromatography (SunFire C18 OBD Prep Column (19×150 mm, 5 m), mobile phase A: water (0.1% FA), mobile phase B: CH3CN, flow rate: 25 mL / min, gradient: isocratic (38→61), wavelength: 254 nm / 220 nm, RT1 (min): 6.83) to obtain the title compound (0.2491 g, 49%) as a pale yellow oil.

[0480] MS(ESI)m / z=411.1[M+H] +

[0481] 1 ¹H NMR (300MHz, chloroform-d) δ 7.43-7.29 (m,1H), 7.28-7.14 (m,2H), 7.11-6.99 (m,1H), 5.35 (m,1H), 3.83-3.63 (m,13H), 3.55-3.53 (m,1H), 3.39-3.34 (m,2H), 2.72-2.36 (m,2H), 2.18-2.10 (m,1H), 2.01-1.72 (m,3H)

[0482] Example 21: Synthesis of Ligand 23 N-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-2-(2-chlorophenyl)pyrrolidine-1-carboxamide [ka] To a stirred solution of 2-[(8-azido-3,6-dioxaocto-1-yl)oxy]ethane-1-amine (300 mg, 1.375 mmol) in DCM (6 mL), disuccinimidyl carbonate (422.53 mg, 1.649 mmol) was added in fractions at 0°C, and the mixture was stirred at room temperature under a nitrogen atmosphere for 4 hours. Subsequently, DIEA (44.06 mg, 0.341 mmol) and 2-(2-chlorophenyl)tetrahydropyrrole (247.71 mg, 1.364 mmol) were added dropwise at 0°C, and the resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The reaction was quenched by adding a saturated NaHCO3 solution at 0°C, and the mixture was extracted with ethyl acetate. The combined organic extracts were washed with water and brine, dried over Na2SO4, filtered, concentrated, and purified by preparative HPLC (SunFire C18 OBD Prep Column (19×150mm, 5m), mobile phase A: water (0.1% FA), mobile phase B: ACN, flow rate: 25 mL / min, gradient: isocratic (37→47), wavelength: 254 nm / 220 nm, RT1 (min): 8.26) to obtain the title compound (180 mg, 31%) as a colorless oil.

[0483] MS(ESI)m / z=426.1[M+H] +

[0484] 1 ¹H NMR (300MHz, chloroform-d) δ 7.36-7.24 (m,1H), 7.20-7.15 (m,1H), 7.15-7.07 (m,2H), 5.16-5.00 (m,1H), 4.01 (s,1H), 3.71-3.51 (m,8H), 3.50-3.38 (m,4H), 3.37-3.23 (m,5H), 2.43-2.26 (m,1H), 1.89-1.71 (m,3H)

[0485] In summary, the following NMDA ligands were prepared. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0486] Preparation of ligand-conjugate oligonucleotides Exemplary ligand-conjugate oligonucleotides that fall within the scope of this disclosure may be synthesized according to the general procedure described below.

[0487] In some cases, one ligand (ligand A in the general procedure below) is conjugated to the 5' end of the oligonucleotide.

[0488] In some cases, two identical ligands (ligand A and ligand A) are conjugated to the 5' and 3' ends of the oligonucleotide.

[0489] In some cases, two different ligands (ligand A and ligand B) are conjugated to the 5' and 3' ends of the oligonucleotide.

[0490] In some cases, one ligand (ligand A) is conjugated to the 3' end of the oligonucleotide.

[0491] Example 22: Ligand conjugated to the 5' end of the sense chain [ka]

[0492] Step 1: 5'-DBCO functionalized sense chain Sodium phosphate buffer (10% (V / V), 1M, pH 7) and acetonitrile (20%-50% (V / V)) were added to an aqueous solution of 5'-amine-functionalized sense chains. Subsequently, a solution of DBCO-NHS (1.5-3 equivalents) in DMSO or acetonitrile was added, and the reaction was monitored by LC-MS and HPLC. Once complete, either precipitate was removed by centrifugation, the aqueous solution was purified by reverse-phase HPLC, dried by lyophilization, and the dried 5'-DBCO-functionalized sense chains were reconstituted with RNase-free water.

[0493] Step 2: 5'-ligand conjugate sense chain A solution of ligand A-N3 (2 equivalents) in DMSO or THF was added to a solution of a 5'-DBCO modified sense chain (1 equivalent), and the reaction was monitored by HPLC and LC-MS. Once complete, the 5'-conjugate sense chain was purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter.

[0494] Example 23: Ligand conjugated to the 5' end of the sense chain [ka]

[0495] Step 1: 5'-DBCO functionalized sense chain Add sodium phosphate buffer (10% (V / V), 1M, pH 7) to an aqueous solution of 5'-(C6-SS-C6)-mC functionalized sense chain. Add tris(2-carboxyethyl)phosphine hydrochloride (TCEP) (25 equivalents) in water (pH 7), and monitor the reaction by HPLC and LC-MS. Once complete, remove excess TCEP with sodium phosphate buffer (100 mM, pH 7, 3×) by molecular weight cutoff. Add a solution of DBCO-MAL (3 equivalents) in DMSO, and monitor the reaction by LC-MS and HPLC. Once complete, remove any solids by centrifugation, purify the solution by reverse-phase HPLC, dry by lyophilization, and reconstitute the dried product with RNase-free water.

[0496] Step 2: 5'-ligand conjugate sense chain A solution of ligand A-N3 (3 equivalents) in DMSO or THF is added to a solution of a 5'-DBCO-functionalized sense chain (1 equivalent), and the reaction is monitored by HPLC and LC-MS. Once complete, the 5'-conjugate sense chain is purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter.

[0497] Example 24: Ligand conjugated to the 5' end of the sense chain [ka]

[0498] Step 1: 5'-DBCO functionalized sense chain 10-(6-oxo-6-(dibenzo[b,f]azacycloocto-4-yn-1-yl)-capramido-N-ethyl)-O-triethylene glycol-1-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite (5'-DBCO-TEG phosphoramidite, purchased from Glen Research, 10-1941-02E) was coupled to the 5' end of the sense chain under standard phosphoramidite coupling conditions.

[0499] Step 2: 5'-ligand conjugate sense chain A solution of ligand A-N3 (2 equivalents) in DMSO is added to a solution of a 5'-DBCO-modified sense chain (1 equivalent), and the reaction is monitored by HPLC and LC-MS. Once complete, the 5'-conjugate sense chain is purified by reverse-phase HPLC or molecular weight cutoff using an Amicon® Ultra-15 centrifugal filter.

[0500] Example 25: Bis-homo-3',5'-ligand conjugated to a sense chain [ka]

[0501] Step 1: 3',5'-bis-DBCO modified sense chain Sodium phosphate buffer (10% (V / V), 1M, pH 7) and acetonitrile (20%-50% (V / V)) were added to an aqueous solution of 3',5' amine-functionalized sense chains. Subsequently, a solution of DBCO-NHS (3 equivalents) in DMSO or CH3CN was added, and the reaction was monitored by LC-MS and HPLC. Upon completion, the product was purified by reverse-phase HPLC, dried by lyophilization, and reconstituted with RNase-free water.

[0502] Step 2: 3',5'-bis-conjugate sense chain A solution of 3',5'-bis-DBCO modified sense chain (1 equivalent) was added to a solution of ligand A-N3 (3 equivalents) in DMSO or CH3CN, and the reaction was monitored by HPLC and LC-MS. Once complete, the 3',5'-bis conjugate sense chain was purified by reverse-phase HPLC, dried by lyophilization, reconstituted with RNase-free water, and desalted using an Amicon® Ultra-15 centrifugal filter (3K, 5 passes).

[0503] Example 26: Bis-homo-3',5'-ligand conjugated to a sense chain [ka]

[0504] Step 1: 3',5'-bis-DBCO modified sense chain Add sodium phosphate buffer (10% (V / V), 1M, pH 7) to an aqueous solution of 5',3'-bis(C6-SS-C6)-mC functionalized sense chain. Add tris(2-carboxyethyl)phosphine hydrochloride (TCEP) (25 equivalents) in water (pH 7), and monitor the reaction by HPLC and LC-MS. Once complete, remove excess TCEP with sodium phosphate buffer (100 mM, pH 7, 3×) by MWCO. Add a solution of DBCO-MAL (3 equivalents) in DMSO, and monitor the reaction by LC-MS and HPLC. Once complete, remove any solids by centrifugation, purify the solution by reverse-phase HPLC, dry by lyophilization, and reconstitute the dried bis-DBCO modified sense chain with RNase-free water.

[0505] Step 2: 3',5'-bis-conjugate sense chain A solution of ligand A-N3 (3 equivalents) in DMSO or THF is added to a solution of a 5',3'-bis-DBCO functionalized sense chain (1 equivalent), and the reaction is monitored by HPLC and LC-MS. Once complete, the bis-homo-5'-,3' conjugate sense chain is purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter.

[0506] Example 27: Bis-homo-3',5'-ligand conjugate sense chain [ka]

[0507] Step 1: 5'-DBCO / 3'-(C6-SS-C6)-mC functionalized sense strand Sodium phosphate buffer (10% (V / V), 1M, pH 7) and acetonitrile (20%-50% (V / V)) were added to an aqueous solution of 5'-amine-functionalized sense chains. Subsequently, a solution of DBCO-NHS (1.5-3 equivalents) in DMSO or acetonitrile was added, and the reaction was monitored by LC-MS and HPLC. Once complete, either precipitate was removed by centrifugation, and the aqueous solution was purified by reverse-phase HPLC. The product fractions were combined and dried by lyophilization, and the dried N-DBCO-modified sense chains were reconstituted with RNase-free water for step 2.

[0508] Step 2: 5',3'-BisDBCO functionalized sense chain Sodium phosphate buffer (10% (V / V), 1M, pH 7) was added to an aqueous solution of 5'-DBCO / 3'-(C6-SS-C6)-mC functionalized sense chains. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) (25 equivalents) in water (pH 7) was added, and the reaction was monitored by HPLC and LC-MS. Once complete, excess TCEP was removed by MWCO with sodium phosphate buffer (100 mM, pH 7, 3×). A solution of DBCO-MAL (3 equivalents) in DMSO was added, and the reaction was monitored by LC-MS and HPLC. Once complete, any solids were removed by centrifugation, the solution was purified by reverse-phase HPLC, dried by lyophilization, and the dried bis-DBCO modified sense chains were reconstituted with RNase-free water for step 3.

[0509] Step 3: Bis-homo-5',3'-ligand conjugate sense chain A solution (1 equivalent) of a 5'-,3'-bis-DBCO-functionalized sense chain was added to a solution (3 equivalents) of ligand A-N3 in DMSO or THF, and the reaction was monitored by HPLC and LC-MS. Once complete, the bis-homo-5'-,3' conjugate sense chain was purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter. The product was confirmed by HPLC and LC-MS.

[0510] Example 28: Bis-homo-3',5'-ligand conjugate sense chain [ka]

[0511] Step 1: 5'-(C6-SS-C6)-mC / 3'-DBCO functionalized sense strand Sodium phosphate buffer (10% (V / V), 1M, pH 7) and acetonitrile (20%-50% (V / V)) were added to an aqueous solution of 5'-amine-functionalized sense chains. Subsequently, a solution of DBCO-NHS (1.5-3 equivalents) in DMSO or acetonitrile was added, and the reaction was monitored by LC-MS and HPLC. Once complete, either precipitate was removed by centrifugation, and the aqueous solution was purified by reverse-phase HPLC. The product fractions were combined and dried by lyophilization, and the dried N-DBCO-modified sense chains were reconstituted with RNase-free water for step 2.

[0512] Step 2: 5',3'-BisDBCO functionalized sense chain Sodium phosphate buffer (10% (V / V), 1M, pH 7) was added to an aqueous solution of 5'-DBCO / 3'-(C6-SS-C6)-mC functionalized sense chains. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) (25 equivalents) in water (pH 7) was added, and the reaction was monitored by HPLC and LC-MS. Once complete, excess TCEP was removed by MWCO with sodium phosphate buffer (100 mM, pH 7, 3×). A solution of DBCO-MAL (3 equivalents) in DMSO was added, and the reaction was monitored by LC-MS and HPLC. Once complete, any solids were removed by centrifugation, the solution was purified by reverse-phase HPLC, dried by lyophilization, and the dried bis-DBCO modified sense chains were reconstituted with RNase-free water for step 3.

[0513] Step 3: Bis-homo-5',3'-ligand conjugate sense chain A solution (1 equivalent) of a 5'-,3'-bis-DBCO-functionalized sense chain was added to a solution (3 equivalents) of ligand A-N3 in DMSO or THF, and the reaction was monitored by HPLC and LC-MS. Once complete, the bis-homo-5'-,3' conjugate sense chain was purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter. The product was confirmed by HPLC and LC-MS.

[0514] Example 29: Bis-hetero-3',5'-ligand conjugate sense chain [ka]

[0515] Step 1: 5'-Conjugate 3'-(C6-SS-C6)-mC Functionalized Sense Chain An aqueous solution of ligand A-N3 (2 equivalents) in DMSO was added to an aqueous solution of a 5'-DBCO-modified sense chain (1 equivalent; see above for preparation), and the reaction was monitored by HPLC and LC-MS. Once complete, the 5'-conjugate sense chain was purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter.

[0516] Step 2: 5'-Conjugate 3'-DBCO Modified Sense Chain To a solution of 5'-conjugate 3'(C6-SS-C6)-mC functionalized sense chain (1 equivalent) in water, sodium phosphate buffer (10% (V / V), 1M, pH 7) was added. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP, 25 equivalents) in water (pH 7) was added, and the reaction was monitored by HPLC and LC-MS. Once complete, excess TCEP was removed by MWCO with sodium phosphate buffer (100 mM, pH 7, 3×). A solution of DBCO-MAL (3 equivalents) in DMSO was added, and the reaction was monitored by HPLC and LC-MS. Once complete, the aqueous solution was purified by reverse-phase HPLC, dried by lyophilization, and the dried 5'-conjugate 3'-DBCO modified sense chain was reconstituted with sodium phosphate buffer (100 mM) for step 3.

[0517] Step 3: Bis-hetero-3',5'-ligand conjugate sense chain An aqueous solution of ligand B-N3 (2 equivalents) in DMSO was added to an aqueous solution of a 5'-conjugate 3'-DBCO-functionalized sense chain (1 equivalent), and the reaction was monitored by HPLC and LC-MS. After completion, the 5'-,3'-conjugate sense chain was purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter.

[0518] Example 30: Bis-hetero-3',5'-ligand conjugate sense chain [ka]

[0519] Step 1: 5'-(C6-SS-C6)-mC, 3'-conjugate sense chain An aqueous solution of ligand A-N3 (2 equivalents) in DMSO was added to an aqueous solution of a 3'-DBCO-modified sense chain (1 equivalent; see above for preparation), and the reaction was monitored by HPLC and LC-MS. Once complete, the 3'-conjugate sense chain was purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter.

[0520] Step 2: 5'-DBCO, 3'-Conjugate sense chain To a solution of 5'-conjugate 3'-(C6-SS-C6)-mC functionalized sense chain (1 equivalent) in water, sodium phosphate buffer (10% (V / V), 1M, pH 7) was added. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP, 25 equivalents) in water (pH 7) was added, and the reaction was monitored by HPLC and LC-MS. Once complete, excess TCEP was removed by MWCO with sodium phosphate buffer (100 mM, pH 7, 3×). A solution of DBCO-MAL (3 equivalents) in DMSO was added, and the reaction was monitored by HPLC and LC-MS. Once complete, the aqueous solution was purified by reverse-phase HPLC, dried by lyophilization, and the dried 5'-conjugate 3'-DBCO modified sense chain was reconstituted with sodium phosphate buffer (100 mM) for step 3.

[0521] Step 3: Bis-hetero-3',5'-ligand conjugate sense chain An aqueous solution of ligand B-N3 (2 equivalents) in DMSO was added to an aqueous solution of a 5'-conjugate 3'-DBCO-functionalized sense chain (1 equivalent), and the reaction was monitored by HPLC and LC-MS. After completion, the 5'-,3'-conjugate sense chain was purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter.

[0522] Example 31: Ligand conjugated to the 3' end of the sense chain [ka]

[0523] Step 1: 3'-DBCO modified sense chain To an aqueous solution of 3'-(C6-SS-C6)-mC functionalized sense chains, sodium phosphate buffer (10% (V / V), 1M, pH 7) was added. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP, 25 equivalents) in water (pH 7) was added, and the reaction was monitored by HPLC and LC-MS. Upon completion, excess TCEP was removed by MWCO with sodium phosphate buffer (100 mM, pH 7, 3×). A solution of DBCO-MAL (3 equivalents) in DMSO was added, and the reaction was monitored by HPLC and LC-MS. Upon completion, any solids were removed by centrifugation, the solution was purified by reverse-phase HPLC, dried by lyophilization, and the dried 3'-DBCO modified sense chains were reconstituted with 100 mM sodium phosphate buffer for step 2.

[0524] Step 2: 3'-ligand conjugate sense chain An aqueous solution of 3'-DBCO-functionalized sense chain (1 equivalent) was mixed with a solution of ligand A-N3 (3 equivalents) in DMSO, and the reaction was monitored by HPLC and LC-MS. Once complete, the 3'-conjugate sense chain was purified by reverse-phase HPLC or molecular weight cutoff (3K, 5×) using an Amicon® Ultra-15 centrifugal filter.

[0525] Example 32: Ligand conjugated to the 3' end of the sense chain [ka]

[0526] Step 1: 3'-DBCO-functionalized sense chain Add sodium phosphate buffer (10% (V / V), 1M, pH 7) and acetonitrile (20%-50% (V / V)) to an aqueous solution of 3'-amine-functionalized sense chains. Then, add a solution of DBCO-NHS (1.5-3 equivalents) in DMSO or acetonitrile, and monitor the reaction by LC-MS and HPLC. Once complete, remove any precipitate by centrifugation, purify the aqueous solution by reverse-phase HPLC, dry by lyophilization, and reconstitute the dried DBCO-modified sense chains with RNase-free water.

[0527] Step 2: 3'-ligand conjugate sense chain A solution of ligand A-N3 (2 equivalents) in DMSO or THF was added to a solution of 3'-DBCO-modified sense strand (1 equivalent), and the reaction was monitored by HPLC and LC-MS. Once complete, the 3'-conjugate sense strand was purified by reverse-phase HPLC or molecular weight cutoff (3K, 5 times) using an Amicon® Ultra-15 centrifugal filter.

[0528] General procedure for chain preparation Example 33: Chain synthesis Oligonucleotide chains were synthesized on a solid phase using the Oligopilot100 oligonucleotide synthesizer (Cytiva Life Sciences). Solid supports (CPG, 80-90 μmol / g, 500A, LGC-Biosearch Technologies (Petaluma, CA)) were packed in 150-300 μmol scales. RNA and 2'-modified RNA phosphoramidites were purchased from Hongene Biotech (Union City, CA).

[0529] The 2'-O-methylphosphoramidites used were as follows: 5'-O-(4,4'-dimethoxytrityl)-N 6 -Benzoyl-2'-O-methyl-adenosine-3'-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite 5'-O-(4,4'-dimethoxytrityl)-N4- Acetyl-2'-O-methylcytidine-3'-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite 5'-O-(4,4'-dimethoxytrityl)-N 2 -Isobutyl-2'-O-methyl-guanosine-3'-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite 5'-O-(4,4'-dimethoxytrityl)-2'-O-methyluridine-3'-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite

[0530] The 2'-fluorophosphoramidites used were as follows: 5'-O-(4,4'-dimethoxytrityl)-N 6 -Benzoyl-2'-fluoroadenosine-3'-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite 5'-O-(4,4'-dimethoxytrityl)-N 4- Acetyl-2'-fluorocytidine-3'-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite 5'-O-(4,4'-dimethoxytrityl)-N 2 -Isobutyl-2'-fluoroguanosine-3'-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite 5'-O-(4,4'-dimethoxytrityl)-2'-fluorouridine-3'-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite

[0531] To form phosphorothioate bonds, 3-((dimethylamino-methylidene)amino)-3H-1,2,4-dithiazol-3-thione (DDTT, 0.1 M solution, Chemgenes (Wilmington, MA)) was used for 4-6 minutes. To form phosphodiester bonds, a solution of pyridine / I2O in water (0.05 M, Sigma Aldrich (St Louis, MO)) was used. After oxidation / sulfurization, any unreacted chains bound to the CPG were acetylated using 20% ​​n-methylimidazole in acetonitrile and a mixture of 40% acetic anhydride in 60% lutidine in acetonitrile (Sigma Aldrich (St Louis, MO)).

[0532] Phosphoamidite was dissolved in anhydrous acetonitrile (0.2 M), molecular sieves (4A) were added, and the mixture was set overnight (Sigma Aldrich (St. Louis, MO)). 5-(ethylthio)-1H-tetrazole (ETT, 0.6 M in acetonitrile, Sigma Aldrich) was used as the activator solution for the oligonucleotide chain. Coupling times were 6 minutes and 3.0 equivalents for each step. Prior to coupling, the oligonucleotides bound to the support were treated with a solution of dichloroacetic acid in dichloromethane (3% Deblock, Sigma Aldrich) and washed with anhydrous acetonitrile.

[0533] Example 34: Cleavage and deprotection of support-bound oligomers After the completion of solid-phase synthesis, the support was treated with AMA solution (a 1:1 volume solution of NH4OH and CH3NH2 (Fisher Scientific, Spectrum Chemicals)) for 20 minutes at 65°C. Subsequently, the solution was evaporated. Before purification, in-process analysis was performed using analytical HPLC and LC-MS to determine crude purity, identify the target mass, and monitor the completion of deprotection.

[0534] Example 35: LC-MS method A Waters XBridge Oligonucleotide BEH C18 column (130 Å, 2.5 μm, 2.1 mm × 50 mm (P / N 186003952)) was used with a buffer solution (400 mM HFIP + 15 mM TEA (buffer A) and 100% methanol (buffer B)) in a gradient of 15 → 40% or 50 → 75% of buffer B, at 15 CV, 70°C, and a flow rate of 1 mL / min.

[0535] Example 36: Concentration by tangential flow filtration (TFF) The crude oligo is concentrated using the Pall Minimate EVO System (product ID: OAPMPUNV), which uses Pall Minimate TFF cassette capsules with a 3k 0mega membrane.

[0536] Example 37: Purification Purification was performed using reverse-phase HPLC (Waters XBridge Prep C18, OBD 5μm, 250×19mm (P / N:186004021)). The buffer solution mixture consisted of 100 mM TEAA, 5% ACN (pH 7.0) (Buffer A) and 1:1 acetonitrile and methanol (Buffer B). The gradient was 5→30% or 30→60% of Buffer B over 60 minutes at 60°C and a flow rate of 20 mL / min.

[0537] After purification, the fractions were analyzed by reverse-phase UPLC. The column used was Waters ACQUITY UPLC oligonucleotide BEH C18 (1.7 μm, 2.1 × 50 mm) (P / N: 186003949). The buffer solution mixture consisted of 100 mM TEAA, 5% ACN (pH 7.0) (Buffer A) and 1:1 acetonitrile and methanol (Buffer B). The gradient was set at 70°C and a flow rate of 1.0 mL / min over 5 minutes, transitioning from 5% to 30% or 30% to 60% of Buffer B. The minimum specification of the purification pool was 85%.

[0538] Example 38: Desalting Once the pool is established, the oligosaccharides are desalted using the Pall Minimate EVO System (Product ID: OAPMPUNV). The cassette used is a 3k Omega membrane (Product ID: OA003C12) along with Pall Minimate TFF capsules. Unpermeated material is collected for direct freeze-drying or annealing.

[0539] Example 39: General procedure for annealing The concentrations of both the sense and antisense strands were determined using Nanodrop. Double-stranded siRNA was prepared by mixing equimolar amounts of sense and antisense strands. The annealing process was monitored using RP-HPLC, a non-denaturing method. After annealing, less than 5% of the antisense strand was present in the double-stranded mixture. The double-stranded concentration was determined by measuring the solution absorbance using Nanodrop.

[0540] Preparation of NMDA ligand-sense chain conjugates Following the process described above, the sense strands targeting target A (regions I and II), target B (regions I, II, and III), and target C were conjugated with ligands 1-23 as described above.

[0541] Example 40: Preparation of Conjugate 1 Following the general procedure described above, ligand 1 was conjugated to the 5' end of an oligosense chain targeting target A-region I.

[0542] The product was prepared with 99% purity and confirmed by HPLC.

[0543] LCMS: m / z: 8098.6 (calculated value 8100.0 g / mol) [ka] In the equation, X is either S or O.

[0544] Example 41: Preparation of Conjugate 2 Following the general procedure described above, ligand 2 was conjugated to the 5' end of an oligosense chain targeting target A-region I.

[0545] The product was prepared with a purity of 98% and confirmed by HPLC.

[0546] LCMS: m / z: 8204.4 (calculated value 8206.0 g / mol) [ka] In the equation, X is either S or O.

[0547] Example 42: Preparation of Conjugate 3 Following the general procedure described above, ligand 4 was conjugated to the 5' end of an oligosense chain targeting target A-region I.

[0548] The product was prepared with a purity of 92% and confirmed by HPLC.

[0549] LCMS:m / z:8147.2 (calculated value 8148.9g / mol) [ka] In the equation, X is either S or O.

[0550] Example 43: Preparation of Conjugate 4 Following the general procedure described above, ligand 5 was conjugated to the 5' end of an oligosense chain targeting target A-region I.

[0551] The product was prepared with a purity of 98% and confirmed by HPLC.

[0552] LCMS:m / z:8177.3 (calculated value 8178.9g / mol) [ka] In the equation, X is either S or O.

[0553] Example 44: Preparation of Conjugate 5 Following the general procedure described above, ligand 6 was conjugated to the 5' end of an oligosense chain targeting target A-region I.

[0554] The product was prepared with a purity of 98% and confirmed by HPLC.

[0555] LCMS:m / z:8154.2 (calculated value 8155.9g / mol) [ka] In the equation, X is either S or O.

[0556] Example 45: Preparation of Conjugate 6 Following the general procedure described above, ligand 9 was conjugated to the 5' end of an oligosense chain targeting target A-region I.

[0557] The product was prepared with a purity of 97% and confirmed by HPLC.

[0558] LCMS: m / z: 8162.3 (calculated value 8164.0 g / mol) [ka] In the equation, X is either S or O.

[0559] Example 46: Preparation of Conjugate 7 Following the general procedure described above, ligand 1 was conjugated to the 5' end of an oligosense chain targeting target A-region II.

[0560] The product was prepared with a purity of 95% and confirmed by HPLC.

[0561] LCMS: m / z: 8555.7 (calculated value 8557.4 g / mol)

[0562] Example 47: Preparation of Conjugate 8 Following the general procedure described above, ligand 2 was conjugated to the 5' end of an oligosense chain targeting target A-region II.

[0563] The product was prepared with a purity of 98% and confirmed by HPLC.

[0564] LCMS: m / z: 8661.6 (calculated value 8663.4 g / mol)

[0565] Example 48: Preparation of Conjugate 9 Following the general procedure described above, ligand 2 was conjugated to the 5' end of an oligosense chain targeting target B-region I.

[0566] The product was prepared with a purity of 92% and confirmed by HPLC.

[0567] LCMS: m / z: 8406.6 (calculated value 8408.1 g / mol)

[0568] Example 49: Preparation of Conjugate 10 Following the general procedure described above, ligand 2 was conjugated to the 5' end of an oligosense chain targeting target B-region II.

[0569] The product was prepared with a purity of 86% and confirmed by HPLC.

[0570] LCMS: m / z: 8440.4 (calculated value 8442.5 g / mol)

[0571] Example 50: Preparation of Conjugate 11 Following the general procedure described above, ligand 2 was conjugated to the 5' end of an oligosense chain targeting the B-region III.

[0572] The product was prepared with a purity of 94% and confirmed by HPLC. LCMS: m / z: 8356.4 (calculated value 8358.1 g / mol)

[0573] Example 51: Preparation of Conjugate 12 Following the general procedure described above, ligand 1 was conjugated to the 5' end of an oligosense chain targeting the target C-region I.

[0574] The product was prepared with a purity of 95% and confirmed by HPLC.

[0575] LCMS: m / z: 7687.6 (calculated value 7688.7 g / mol)

[0576] Example 52: Preparation of Conjugate 13 Following the general procedure described above, ligand 13 was conjugated to the 5' end of an oligosense chain targeting target A-region I.

[0577] The product was prepared with a purity of 89% and confirmed by HPLC.

[0578] LCMS: m / z: 8797.7 (calculated value 8799.5 g / mol) [ka] In the equation, X is either S or O.

[0579] Example 53: Preparation of Conjugate 14 Following the general procedure described above, ligand 14 was conjugated to the 5' end of an oligosense chain targeting target A-region II.

[0580] The product was prepared with a purity of 90% and confirmed by HPLC.

[0581] LCMS: m / z: 8660.8 (calculated value 8662.4 g / mol) [ka] In the equation, X is either S or O.

[0582] Example 54: Preparation of Conjugate 15 Following the general procedure described above, ligand 15 was conjugated to the 5' end of an oligosense chain targeting target A-region II.

[0583] The product was prepared with a purity of 94% and confirmed by HPLC.

[0584] LCMS: m / z: 8660.9 (calculated value 8662.4 g / mol) [ka] In the equation, X is either S or O.

[0585] Example 55: Preparation of Conjugate 16 Following the general procedure described above, ligand 10 was conjugated to the 5' end of an oligosense chain targeting target A-region II.

[0586] The product was prepared with a purity of 90% and confirmed by HPLC.

[0587] LCMS: m / z: 8661.6 (calculated value 8663.1 g / mol) [ka] In the equation, X is either S or O.

[0588] Example 56: Preparation of Conjugate 17 Following the general procedure described above, ligand 7 was conjugated to the 5' end of an oligosense chain targeting target A-region II.

[0589] The product was prepared with a purity of 89% and confirmed by HPLC.

[0590] LCMS: m / z: 8592.0 (calculated value 8593.7 g / mol) [ka] In the equation, X is either S or O.

[0591] Example 57: Preparation of Conjugate 18 Following the general procedure described above, ligand 8 was conjugated to the 5' end of an oligosense chain targeting target A-region II.

[0592] The product was prepared with a purity of 93% and confirmed by HPLC.

[0593] LCMS: m / z: 8592.0 (calculated value 8593.7 g / mol) [ka] In the equation, X is either S or O.

[0594] Example 58: Preparation of Conjugate 19 Following the general procedure described above, ligand 1 was conjugated to the 5' end of an oligosense chain targeting target A-region II.

[0595] The product was prepared with a purity of 98% and confirmed by HPLC.

[0596] LCMS: m / z: 8696.2 (calculated value 8697.8 g / mol) [ka] In the equation, X is either S or O.

[0597] Example 59: Preparation of Conjugate 20 Following the general procedure described above, ligand 2 was conjugated to the 3' end of an oligosense chain targeting target A-region I.

[0598] The product was prepared with a purity of 97% and confirmed by HPLC.

[0599] LCMS: m / z: 8560.1 (calculated value 8561.8 g / mol) [ka] In the equation, X is either S or O.

[0600] Example 60: Preparation of Conjugate 21 Following the general procedure described above, ligand 2 was conjugated to the 5' and 3' ends of an oligosense chain targeting target A-region II.

[0601] The product was prepared with a purity of 85% and confirmed by HPLC.

[0602] LCMS: m / z: 9132.2 (calculated value 9134.0 g / mol) [ka] In the equation, each X is independently either S or O.

[0603] Example 61: Preparation of Conjugate 22 Following the general procedure described above, ligand 19 was conjugated to the 5' and 3' ends of an oligosense chain targeting target A-region I.

[0604] The product was prepared with a purity of 85% and confirmed by HPLC. LCMS:m / z:9087.5 (calculated value 9088.9g / mol) [ka] In the equation, each X is independently either S or O.

[0605] Example 62: Preparation of Conjugate 23 Following the general procedure described above, ligand 20 was conjugated to the 5' and 3' ends of an oligosense chain targeting target A-region I.

[0606] The product was prepared with a purity of 95% and confirmed by HPLC.

[0607] LCMS: m / z: 9079.4 (calculated value 9081.0 g / mol) [ka] In the equation, each X is independently either S or O.

[0608] Example 63: Preparation of Conjugate 24 Following the general procedure described above, ligand 18 was conjugated to the 5' and 3' ends of an oligosense chain targeting target A-region I.

[0609] The product was prepared with a purity of 85% and confirmed by HPLC.

[0610] LCMS: m / z: 9167.6 (calculated value 9169.2 g / mol) [ka] In the equation, each X is independently either S or O.

[0611] Example 64: Preparation of Conjugate 25 Following the general procedure described above, ligand 17 was conjugated to the 5' and 3' ends of an oligosense chain targeting target A-region I.

[0612] The product was prepared with a purity of 93% and confirmed by HPLC.

[0613] LCMS: m / z: 9037.4 (calculated value 9039.2 g / mol) [ka] In the equation, each X is independently either S or O.

[0614] Example 65: Preparation of Conjugate 26 Following the general procedure described above, ligand 21 was conjugated to the 5' and 3' ends of an oligosense chain targeting target A-region I.

[0615] The product was prepared with a purity of 96% and confirmed by HPLC.

[0616] LCMS: m / z: 9028.4 (calculated value 9029.4 g / mol) [ka] In the equation, each X is independently either S or O.

[0617] Example 66: Preparation of Conjugate 27 Following the general procedure described above, ligand 22 was conjugated to the 5' and 3' ends of an oligosense chain targeting target A-region I.

[0618] The product was prepared with a purity of 93% and confirmed by HPLC.

[0619] LCMS: m / z: 8998.7 (calculated value 8999.7 g / mol) [ka] In the equation, each X is independently either S or O.

[0620] Example 67: Preparation of Conjugate 28 Following the general procedure described above, ligand 23 was conjugated to the 5' and 3' ends of an oligosense chain targeting target A-region I.

[0621] The product was prepared with a purity of 93% and confirmed by HPLC.

[0622] LCMS: m / z: 9028.5 (calculated value 9029.8 g / mol) [ka] In the equation, each X is independently either S or O.

[0623] Example 68: Preparation of Conjugate 29 Following the general procedure described above, ligand 2 was conjugated to the 5' and 3' ends of an oligosense chain targeting the target C-region II.

[0624] The product was prepared with 99% purity and confirmed by HPLC.

[0625] LCMS:m / z:9405.2 (calculated value 9406.9g / mol)

[0626] Example 69: Preparation of Conjugate 30 Following the general procedure described above, ligand 2 was conjugated to the 5' and 3' ends of an oligosense chain targeting the C-region III.

[0627] The product was prepared with 99% purity and confirmed by HPLC.

[0628] LCMS:m / z:9444.3 (calculated value 9445.9g / mol)

[0629] Example 70: Preparation of Conjugate 31 Following the general procedure described above, ligand 2 was conjugated to the 5' and 3' ends of an oligosense chain targeting the target C-region IV.

[0630] The product was prepared with a purity of 97% and confirmed by HPLC.

[0631] LCMS: m / z: 9459.3 (calculated value 9460.8 g / mol)

[0632] Example 71: Preparation of Conjugate 32 Following the general procedure described above, ligand 2 was conjugated to the 5' and 3' ends of an oligosense chain targeting the target C-region V.

[0633] The product was prepared with a purity of 98% and confirmed by HPLC.

[0634] LCMS: m / z: 9451.3 (calculated value 9453.0 g / mol) [ka] In the equation, each X is independently either S or O.

[0635] Example 72: Preparation of Conjugate 33 Following the general procedure described above, ligand 2 was conjugated to the 5' and 3' ends of the oligosense chain targeting target B-region I.

[0636] The product was prepared with a purity of 76% and confirmed by HPLC.

[0637] LCMS:m / z:9524.4 (calculated value 9526.3g / mol)

[0638] Example 73: Preparation of Conjugate 34 Following the general procedure described above, ligand 2 was conjugated to the 5' and 3' ends of an oligosense chain targeting target B-region II.

[0639] The product was prepared with a purity of 83% and confirmed by HPLC.

[0640] LCMS: m / z: 9558.4 (calculated value 9560.2 g / mol)

[0641] Example 74: Preparation of Conjugate 35 Following the general procedure described above, ligand 2 was conjugated to the 5' and 3' ends of an oligosense chain targeting the target B-region VI.

[0642] The product was prepared with a purity of 88% and confirmed by HPLC.

[0643] LCMS:m / z:9620.6 (calculated value 9622.3g / mol) [ka] In the equation, each X is independently either S or O.

[0644] In summary, the following conjugates were prepared. [Table 2]

[0645] in vivo animal experiments NMDA conjugate compounds were evaluated in vivo. RNA was recovered from various brain regions as shown below, and the target inhibition rate was measured. [Table 3]

[0646] Example 75: PD study in vivo in human target A transgenic mice NMDA conjugate compounds were evaluated in vivo in a PD study in human target A transgenic mice. These mice received a single vehicle dose or 0.2 mg (10 mg / kg) via intracerebroventricular injection on day 1 (n=3 / group). The mice were observed daily for changes in behavior. Brain regions were collected on day 15, immediately placed in homogenization tubes, rapidly frozen, and then kept at -80°C for gene expression analysis.

[0647] RNA isolation was performed according to the instructions for the RNeasy Micro Kit (Qiagen catalog number 74004). After RNA isolation, the 96-well plate was placed on ice while preparing the qRT-PCR reaction mixture. 2 μl of RNA was added to the reaction mixture, which contained 5 μl of TaqMan Fast Virus 1-Step Master Mix (Thermo Fisher #44444432), 1 μl of Target A TaqMan Gene Expression Assay (Thermo Fisher), 1 μl of mouse GAPDH (VIC) TaqMan Gene Expression Assay (Thermo Fisher: Mm99999915_g1, VIC), and 11 μl of RT-PCR grade nuclease-free water in a 96-well MicroAmp Optical (0.2 mL) plate. qPCR was performed using a QuantStudio3 qPCR instrument with cycles of 50°C for 1 minute, 95°C for 20 seconds, 95°C for 15 seconds for 40 cycles, and 60°C for 1 minute. The results are shown in the table below as target A inhibition rate (percentage) compared to the vehicle control. [Table 4]

[0648] Example 76: PD study in vivo in human target B transgenic mice NMDA conjugate compounds were evaluated in vivo in a PD (Patient Degeneration) study in human target B transgenic mice. These mice received a single vehicle dose or 0.2 mg (10 mg / kg) via intracerebroventricular injection on day 1 (n=3 / group). The mice were observed daily for changes in behavior. Brain regions were collected on day 15, immediately placed in homogenization tubes, rapidly frozen, and then kept at -80°C for gene expression analysis.

[0649] RNA isolation was performed according to the instructions for the RNeasy Micro Kit (Qiagen catalog number 74004). After RNA isolation, the 96-well plate was placed on ice while preparing the qRT-PCR reaction mixture. 2 μl of RNA was added to the reaction mixture, which contained 5 μl of TaqMan Fast Virus 1-Step Master Mix (Thermo Fisher #44444432), 1 μl of Target B TaqMan Gene Expression Assay (Thermo Fisher), 1 μl of mouse GAPDH (VIC) TaqMan Gene Expression Assay (Thermo Fisher: Mm99999915_g1, VIC), and 11 μl of RT-PCR grade nuclease-free water in a 96-well MicroAmp Optical (0.2 mL) plate. qPCR was performed using a QuantStudio3 qPCR instrument with cycles of 50°C for 1 minute, 95°C for 20 seconds, 95°C for 15 seconds for 40 cycles, and 60°C for 1 minute. The results are shown in the table below as target B inhibition rate (percentage) compared to the vehicle control. [Table 5]

[0650] Example 77: In vivo PD study in rats NMDA conjugate compounds were evaluated in vivo in rat PD studies. Mice were administered a single vehicle dose or 0.9 mg (3 mg / kg) by intracisional cisterna magna (ICM) injection on day 1 (n=3 / group). Mice were observed daily for changes in behavior. Brain regions were collected on day 15, immediately placed in homogenization tubes, rapidly frozen, and kept at -80°C for gene expression analysis.

[0651] RNA isolation was performed according to the instructions for the RNeasy Micro Kit (Qiagen catalog number 74004). After RNA isolation, the 96-well plate was placed on ice while preparing the qRT-PCR reaction mixture. 2 μl of RNA was added to the reaction mixture, which contained 5 μl of TaqMan Fast Virus 1-Step Master Mix (Thermo Fisher #44444432), 1 μl of Rad Target C TaqMan Gene Expression Assay (Thermo Fisher), 1 μl of ACTB (VIC) TaqMan Gene Expression Assay (Thermo Fisher: Rn00667869_m1, VIC), and 11 μl of RT-PCR grade nuclease-free water in a 96-well MicroAmp Optical (0.2 mL) plate. qPCR was performed using a QuantStudio3 qPCR instrument with cycles of 50°C for 1 minute, 95°C for 20 seconds, 95°C for 15 seconds for 40 cycles, and 60°C for 1 minute. The results are shown in the table below as the target C inhibition rate (percentage) compared to the vehicle control. [Table 6]

[0652] Embedding by reference The contents of all references cited throughout this application (including reference documents, issued patents, published patent applications and concurrently pending patent applications) are expressly incorporated herein by reference in their entirety.

[0653] Equal parts Those skilled in the art can recognize or determine, by conventional experimentation, many of the equivalents of the specific embodiments described herein. Such equivalents are intended to be included in the following claims.

[0654] Embodiment Additional embodiments include the following:

[0655] Embodiment P1. A compound comprising the structure of the following formula (I) or a salt thereof, [ka] During the ceremony, [ka] However, it is an N-methyl-D-aspartate (NMDA) receptor ligand, Each of L1, L2, L3, and L4 is either an independent linker, a link, or absent. Y is a bond or -C(=O)-, R 1 The compound is one or more oligonucleotides, protecting groups, small molecules, proteins, antibodies, peptides, or combinations thereof.

[0656] Embodiment P2. The compound of Embodiment P1 o...

Claims

1. A compound comprising the structure of the following formula (I) or a salt thereof, 【Chemistry 375】 During the ceremony, 【Transformation 376】 However, it is an N-methyl-D-aspartate (NMDA) receptor ligand, L １ , L ２ , L ３ and L ４ Each of them is independent, a linker, a connector, or does not exist. Y is a bond or -C (=O)-, R １ The compound is one or more oligonucleotides, protecting groups, small molecules, proteins, antibodies, peptides, or combinations thereof.

2. The compound or a salt thereof according to claim 1, wherein the NMDA receptor ligand is an NMDA receptor agonist.

3. The compound or a salt thereof according to claim 1, wherein the NMDA receptor ligand is an NMDA receptor antagonist.

4. The NMDA receptor ligand is 【Chemical 377】 A compound or salt thereof according to claim 1, selected from the group consisting of anti-NMDA receptor antibodies and derivatives thereof.

5. The aforementioned compound is given by the following formula (II) 【Chemistry 378】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

6. The aforementioned compound is given by the following formula (II-a) 【Chemistry 379】 The compound or a salt thereof according to claim 5, comprising the structure of a salt thereof.

7. The compound is of the following formula (III), 【Chemical 380】 In the formula, R ２ However, the above formula is hydrogen, halogen, -OH or -OMe, The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

8. The aforementioned compound is given by the following formula (III-e) 【Chemistry 381】 The compound or a salt thereof according to claim 7, comprising the structure of a salt thereof.

9. The aforementioned compound is given by the following formula (III-f) 【Chem.382】 The compound or a salt thereof according to claim 7, comprising the structure of a salt thereof.

10. The aforementioned compound is given by the following formula (III-g) 【Chemistry 383】 The compound or a salt thereof according to claim 7, comprising the structure of a salt thereof.

11. The aforementioned compound is given by the following formula (III-h) 【Chemical 384】 A compound or salt thereof according to claim 7, comprising the structure of the compound or salt thereof.

12. The aforementioned compound is given by the following formula (III-i) 【Chem.385】 A compound or salt thereof according to claim 7, comprising the structure of the compound or salt thereof.

13. The aforementioned compound is given by the following formula (III-j) 【Chemical 386】 A compound or salt thereof according to claim 7, comprising the structure of the compound or salt thereof.

14. The aforementioned compound is given by the following formula (III-k) 【Chemistry 387】 A compound or salt thereof according to claim 7, comprising the structure of the compound or salt thereof.

15. The aforementioned compound is given by the following formula (III-l) 【Chemical 388】 A compound or salt thereof according to claim 7, comprising the structure of the compound or salt thereof.

16. The aforementioned compound is given by the following formula (III-m) 【Chem.389】 A compound or salt thereof according to claim 7, comprising the structure of the compound or salt thereof.

17. The aforementioned compound is given by the following formula (III-n) 【Chemical 390】 A compound or salt thereof according to claim 7, comprising the structure of the compound or salt thereof.

18. The aforementioned compound is given by the following formula (III-o) 【Chemistry 391】 A compound or salt thereof according to claim 7, comprising the structure of the compound or salt thereof.

19. The aforementioned compound is given by the following formula (III-p) 【Chemistry 392】 A compound or salt thereof according to claim 7, comprising the structure of the compound or salt thereof.

20. The aforementioned compound is given by the following formula (IV) 【Chemistry 393】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

21. The aforementioned compound is given by the following formula (IV-a) 【Chem. 394】 The compound or a salt thereof according to claim 20, comprising the structure of a salt thereof.

22. The aforementioned compound is given by the following formula (XIX) 【Chemical 395】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

23. The aforementioned compound is given by the following formula (XIX-a) 【Chemistry 396】 The compound or a salt thereof according to claim 22, comprising the structure of a salt thereof.

24. The aforementioned compound is given by the following formula (V) 【Chemistry 397】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

25. The aforementioned compound is given by the following formula (V-a) 【Chem.398】 The compound or a salt thereof according to claim 24, comprising the structure of a salt thereof.

26. The aforementioned compound is given by the following formula (VI) 【Chem.399】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

27. The aforementioned compound is given by the following formula (VI-a) 【Chemical 400】 The compound or a salt thereof according to claim 26, comprising the structure of the salt thereof.

28. The aforementioned compound is given by the following formula (VI-b) 【Chemical 401】 The compound or a salt thereof according to claim 26, comprising the structure of the salt thereof.

29. The aforementioned compound is given by the following formula (VII) 【Chemical 402】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

30. The aforementioned compound is given by the following formula (VII-a) 【Chemical 403】 The compound or a salt thereof according to claim 29, comprising the structure of a salt thereof.

31. The aforementioned compound is given by the following formula (VIII) 【Chemical 404】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

32. The aforementioned compound is given by the following formula (VIII-a) 【Chemical 405】 The compound or a salt thereof according to claim 31, comprising the structure of a salt thereof.

33. The aforementioned compound is given by the following formula (IX) 【Chemical 406】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

34. The aforementioned compound is given by the following formula (IX-a) 【Chemical 407】 The compound or a salt thereof according to claim 33, comprising the structure of a salt thereof.

35. The aforementioned compound is given by the following formula (X) 【Chemical 408】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

36. The aforementioned compound is given by the following formula (X-a) 【Chemical 409】 The compound or a salt thereof according to claim 35, comprising the structure of a salt thereof.

37. The aforementioned compound is given by the following formula (X-b) 【Chemical 410】 The compound or a salt thereof according to claim 35, comprising the structure of a salt thereof.

38. The aforementioned compound is given by the following formula (XI) 【Chemical 411】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

39. The aforementioned compound is given by the following formula (XI-a) 【Chemical 412】 The compound or a salt thereof according to claim 38, comprising the structure of a salt thereof.

40. The aforementioned compound is given by the following formula (XI-b) 【Chemical 413】 The compound or a salt thereof according to claim 38, comprising the structure of a salt thereof.

41. The aforementioned compound is given by the following formula (XI-c) 【Chemical 414】 The compound or a salt thereof according to claim 38, comprising the structure of a salt thereof.

42. The aforementioned compound is given by the following formula (XII) 【Chemical 415】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

43. The aforementioned compound is given by the following formula (XII-a) 【Chemical 416】 The compound or a salt thereof according to claim 42, comprising the structure of a salt thereof.

44. The aforementioned compound is given by the following formula (XII-b) 【Chemical 417】 The compound or a salt thereof according to claim 42, comprising the structure of a salt thereof.

45. The aforementioned compound is given by the following formula (XIII) 【Chemical 418】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

46. The aforementioned compound is given by the following formula (XIII-a) 【Chemical 419】 The compound or a salt thereof according to claim 45, comprising the structure of a salt thereof.

47. The aforementioned compound is given by the following formula (XIV) 【Chemical 420】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

48. The aforementioned compound is given by the following formula (XIV-a) 【Chemistry 421】 The compound or a salt thereof according to claim 47, comprising the structure of a salt thereof.

49. The aforementioned compound is given by the following formula (XV) 【Chemistry 422】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

50. The aforementioned compound is given by the following formula (XV-a) 【Chemistry 423】 The compound or a salt thereof according to claim 49, comprising the structure of a salt thereof.

51. The aforementioned compound is given by the following formula (XVI) 【Chemistry 424】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

52. The aforementioned compound is given by the following formula (XVI-a) 【Chemical 425】 The compound or a salt thereof according to claim 51, comprising the structure of a salt thereof.

53. The aforementioned compound is given by the following formula (XVII) 【Chemistry 426】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

54. The aforementioned compound is given by the following formula (XVII-a) 【Chemistry 427】 The compound or a salt thereof according to claim 53, comprising the structure of a salt thereof.

55. The aforementioned compound is given by the following formula (XVIII) 【Chemistry 428】 The compound or a salt thereof according to claim 1, comprising the structure of a salt thereof.

56. The aforementioned compound is given by the following formula (XVIII-a) 【Chemistry 429】 The compound or a salt thereof according to claim 55, comprising the structure of a salt thereof.

57. wherein L １ , L ２ , L ３ and L ４ each independently is absent, or is a bond, an optionally substituted alkyl linker, an optionally substituted polyethylene glycol (PEG) linker, an optionally substituted heteroalkyl linker, an optionally substituted heteroaryl linker, a phosphodiester bond or a phosphorothioate bond, a compound or a salt thereof according to any one of claims 1 to 56.

58. In the ceremony, L １ A compound or salt thereof according to any one of claims 1 to 57, wherein the bond is a linkage.

59. In the ceremony, L １ The compound or salt thereof according to any one of claims 1 to 57, wherein the alkyl linker is optionally substituted.

60. In the ceremony, L １ However, C is arbitrarily substituted. １ -C ６ A compound or salt thereof according to any one of claims 1 to 57, which is an alkyl linker.

61. In the ceremony, L １ The compound or salt thereof according to any one of claims 1 to 57, wherein the compound is a C1-C6 alkyl linker substituted with =O.

62. In the ceremony, L １ but, 【Chemistry 430】 A compound or salt thereof according to any one of claims 1 to 57, comprising the structure of [the compound].

63. In the ceremony, L ２ The compound or salt thereof according to any one of claims 1 to 62, wherein the alkyl linker is optionally substituted.

64. In the ceremony, L ２ However, C is arbitrarily substituted. １ -C １５ A compound or salt thereof according to any one of claims 1 to 62, which is an alkyl linker.

65. In the ceremony, L ２ However, C is arbitrarily substituted. ５ -C １２ A compound or salt thereof according to any one of claims 1 to 62, which is an alkyl linker.

66. In the ceremony, L ２ but, 【Chemistry 431】 A compound or salt thereof according to any one of claims 1 to 62, comprising the structure of [the compound].

67. In the ceremony, L ２ The compound or salt thereof according to any one of claims 1 to 62, wherein the PEG linker is optionally substituted.

68. In the ceremony, L ２ However, it is an arbitrarily substituted PEG linker, and the length of the PEG unit is 1, 2, 3, 4, 5, 6, 7 or 8, and the PEG unit is 【Chemistry 432】 A compound or salt thereof according to any one of claims 1 to 62, comprising the structure of [the compound].

69. In the ceremony, L ２ However, it is an arbitrarily substituted PEG linker, and the length of the PEG unit is 3, and the PEG unit is 【Chemistry 433】 A compound or salt thereof according to any one of claims 1 to 62, comprising the structure of [the compound].

70. In the ceremony, L ２ However, it is an arbitrarily substituted PEG linker, and the length of the PEG unit is 4, and the PEG unit is 【Chemistry 434】 A compound or salt thereof according to any one of claims 1 to 62, comprising the structure of [the compound].

71. In the ceremony, L ２ but, 【Chemical 435】 A compound or salt thereof according to any one of claims 1 to 62, comprising the structure of [the compound].

72. In the ceremony, L ２ The compound or salt thereof according to any one of claims 1 to 62, wherein the compound is an optionally substituted heteroalkyl linker.

73. In the ceremony, L ２ but, 【Chemistry 436】 A compound or salt thereof according to claim 72, comprising the structure.

74. In the ceremony, L ３ The compound or salt thereof according to any one of claims 1 to 73, wherein the compound is an optionally substituted heteroaryl linker.

75. In the ceremony, L ３ The compound or salt thereof according to any one of claims 1 to 73, wherein the compound is optionally substituted, partially unsaturated, heterocycloalkyl linker, or optionally unsubstituted, heteroaryl linker.

76. In the ceremony, L ３ but, 【Chemistry 437】 A compound or salt thereof according to claim 74 or 75, comprising the structure of the compound or salt thereof.

77. In the ceremony, L ４ The compound or salt thereof according to any one of claims 1 to 76, wherein the compound is an optionally substituted heteroalkyl linker.

78. The compound or a salt thereof according to claim 77, wherein the heteroalkyl linker is substituted with one or more =O substituents.

79. In the ceremony, L ４ but, 【Chemistry 438】 It includes the structure, The compound or salt thereof according to claim 77 or 78, wherein X is O or S in the formula.

80. In the ceremony, L ４ but, 【Chemistry 439】 It includes the structure, The compound or salt thereof according to claim 77 or 78, wherein X is S or O.

81. In the ceremony, L １ , L ２ , L ３ and L ４ But, together, 【Chemical 440-1】 【Chemistry 440-2】 【Chemistry 440-3】 It includes the structure, A compound or salt thereof according to any one of claims 1 to 80, wherein X is O or S.

82. The aforementioned compound, 【Chemistry 441-1】 【Chemistry 441-2】 【Chemistry 441-3】 【Chemistry 441-4】 【Chemistry 441-5】 A compound or salt thereof according to any one of claims 1 to 81, comprising the structure, wherein X is O or S in the formula.

83. A compound or salt thereof according to any one of claims 79 to 82, wherein X is O in the formula.

84. A compound or salt thereof according to any one of claims 79 to 82, wherein X is S in the formula.

85. In the formula, R １ A compound or salt thereof according to any one of claims 1 to 84, wherein the compound comprises an oligonucleotide.

86. The compound or salt thereof according to claim 85, wherein the oligonucleotide is bonded at its 5' end.

87. The compound or salt thereof according to claim 85, wherein the oligonucleotide is bonded at its 3' end.

88. The compound or salt thereof according to claim 85, wherein the oligonucleotide is bonded at an internal position on the oligonucleotide.

89. The compound or salt thereof according to claim 88, wherein the internal position is a nucleoside bond.

90. In the formula, R １ The compound or salt thereof according to any one of claims 1 to 89, comprising an oligonucleotide conjugated to one or more additional NMDA receptor ligands.

91. The compound or salt thereof according to claim 90, wherein the oligonucleotide is conjugated to two, three, four, or five or more additional NMDA receptor ligands.

92. The compound or salt thereof according to claim 90 or 91, wherein the additional NMDA receptor ligand is conjugated to the oligonucleotide at any of the 5' end of the oligonucleotide, the 3' end of the oligonucleotide, one or more internal positions on the oligonucleotide, or a combination thereof.

93. The compound or a salt thereof according to any one of claims 85 to 92, wherein the oligonucleotide is a modified oligonucleotide.

94. A composition comprising a compound or salt thereof according to any one of claims 1 to 93 and a pharmaceutically acceptable excipient.

95. A method for delivering a therapeutic oligonucleotide to a target brain, comprising administering to the target a compound or salt thereof according to any one of claims 1 to 93, or a composition according to claim 94.

96. The method according to claim 95, wherein the therapeutic oligonucleotide is delivered to one or more brain regions selected from the group consisting of the striatum, cerebellum, brainstem, hippocampus, frontal cortex, and spinal cord.

97. A method for treating or improving a disease, disorder or its symptoms, comprising administering to the subject a compound or salt thereof according to any one of claims 1 to 93, or a composition according to claim 94.

98. The method according to claim 97, wherein the disease, disorder or symptoms thereof are a disease, disorder or symptoms thereof of the central nervous system (CNS).

99. The method according to claim 97 or 98, wherein the disease, disorder or its symptoms are Alzheimer's disease or its symptoms.

100. The method according to any one of claims 97 to 99, wherein the compound or a salt thereof is administered intrathecally to the subject.

101. A method for producing a compound or salt thereof according to any one of claims 1 to 93, comprising one or more compounds and chemical conversions described herein (including Examples 1 to 77).

Images