SARM1 inhibitors

Abstract

This disclosure provides compounds and methods that can be used to inhibit SARM1 and / or treat and / or prevent axonal degeneration.

Description

[0001] background

[0002] Axon degeneration is a hallmark of several neurological disorders, including peripheral neuropathy, traumatic brain injury, and neurodegenerative diseases (see, for example, Gerdts et al., SARM1 activation triggers axon degeneration locally via NAD(+)destruction. Science 348 2016, pp. 453-457 and Krauss et al., (2020) Trends Pharmacol. Sci. 41, 281, each incorporated herein by reference in its entirety). Neurodegenerative diseases and injuries are devastating for both patients and caregivers. In the United States alone, the costs associated with these diseases currently exceed several hundred billion dollars annually. Because the incidence of many of these diseases and conditions increases with age, their prevalence increases rapidly with demographic changes.

[0003] Overview

[0004] This disclosure provides techniques, particularly useful for treating and / or preventing neurodegeneration (e.g., for reducing axonal degeneration). In some embodiments, the provided techniques inhibit SARM1.

[0005] In some embodiments, this disclosure provides certain compounds and / or compositions that can be used in medicine, particularly for the treatment of neurodegeneration (e.g., for reducing axonal degeneration).

[0006] All references to Equation I should also be interpreted as references to Equation II.

[0007] In some embodiments, this disclosure provides compounds having the structure shown in Formula I:

[0008]

[0009] Or its pharmaceutically acceptable salt, wherein:

[0010] Ring A is a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur;

[0011] R 1 The substituted group is selected from a 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, or a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0012] R x Each group is independently selected from halogen, cyano, OR, SR, N(R)2, or optionally substituted groups, wherein the groups are selected from C1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbon rings, 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur;

[0013] Ring B is a saturated 5- to 7-membered heterocycle with a structure And optionally includes another option selected from –NH-, -O- and –NR. 2 - group;

[0014] R is each independently hydrogen or an optionally substituted group, said group being selected from C 1-6 Aliphatic group, 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur, phenyl, and 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur; or:

[0015] The two R groups together with the nitrogen atom to which they are attached form a 3- to 7-membered monocyclic heterocycle with 0-2 additional heteroatoms independently selected from oxygen, nitrogen and sulfur;

[0016] R 2 Each independently represents a halogen, N(R)2, OR, or C. 1-3 Aliphatic groups or –(C 1-3 (Aliphatic group)R 3 ;

[0017] R 3 Each group is an optional substituted group, which is selected from C 1-6 Aliphatic group, 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl, 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, 8- to 10-membered bicyclic saturated, partially unsaturated or aryl carbon ring, 8- to 10-membered bicyclic saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, or 8- to 10-membered bicyclic heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur;

[0018] m is 0, 1, or 2; and

[0019] n is 0, 1, or 2.

[0020] In some embodiments, the provided compounds have the structures of the following formulas: Ia, Iai, Ib, Ibi, Ib-ii, Ic, Ici, Id, Idi, Ie, Iei, If, Ifi, Ig, Igi, Ih, Ihi, Ii, Iii, Ij, Iji, and Ij-ii.

[0021] In one embodiment, this disclosure provides compounds of formula II:

[0022]

[0023] Or its pharmaceutically acceptable salt, wherein:

[0024] R 1 It is a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms selected from oxygen, nitrogen, and sulfur;

[0025] G represents CH and CR. x Or N;

[0026] R x It is a C1-C3 alkyl, halogen, or cyano group;

[0027] X is CH2, NH, N (C1-C3 alkyl) or O;

[0028] Y is C(R) p )2 or NH;

[0029] Z represents a valence bond, CH2, or -CH2CH2-;

[0030] R 2a -(C1-C3 alkyl)R 3 ;

[0031] R 2b It is hydrogen, halogen, C1-C3 alkyl or -(C1-C3 alkyl)R 3 ;

[0032] R p Independently hydrogen, halogen, or NH2;

[0033] R 3 It is a benzene ring or a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms selected from oxygen, nitrogen, and sulfur, wherein the aryl ring or the 5- to 6-membered heteroaryl ring is optionally surrounded by 1 to 2 R atoms. q replace;

[0034] R q It can be halogen, cyano, or -CF3.

[0035] In another embodiment, this disclosure provides a compound that is:

[0036]

[0037] Or its pharmaceutically acceptable salt.

[0038] In another embodiment, this disclosure provides a compound that is:

[0039]

[0040] Or its pharmaceutically acceptable salt.

[0041] In another embodiment, this disclosure provides a compound that is:

[0042]

[0043] Or its pharmaceutically acceptable salt.

[0044] In another embodiment, this disclosure provides a compound that is:

[0045]

[0046] Or its pharmaceutically acceptable salt.

[0047] In some embodiments, one or more compounds of Formula I are provided and / or used in solid form (e.g., crystalline or amorphous form).

[0048] In some embodiments, this disclosure provides compositions comprising and / or delivering a Formula I compound (e.g., in the form described herein), its prodrug, or its active metabolite.

[0049] In some embodiments, this disclosure provides compositions comprising and / or delivering a compound of formula I. In some embodiments, such compositions are pharmaceutical compositions comprising at least one pharmaceutically acceptable carrier, diluent, or excipient.

[0050] In some embodiments, the provided compound reduces or inhibits the binding of SARM1 to NAD+. In some embodiments, the provided compound binds to SARM1 within a pouch containing one or more catalytic residues (e.g., a catalytic cleavage of SARM1).

[0051] In some embodiments, the provided compounds and / or compositions inhibit the activity of SARM1. Alternatively or additionally, in some embodiments, the provided compounds alleviate one or more properties of neurodegeneration. In some embodiments, this disclosure provides methods for treating neurodegenerative diseases or conditions associated with axonal degeneration.

[0052] In some embodiments, one or more compounds and / or compositions as described herein may be used, for example, in pharmaceutical practice. In some embodiments, one or more compounds and / or compositions as described herein may be used, for example, to treat, prevent, or improve axonal degeneration (e.g., one or more of its characteristics or properties). In some embodiments, one or more compounds and / or compositions as described herein may be used, for example, to inhibit axonal degeneration, including axonal degeneration caused by a reduction or depletion of NAD+. In some embodiments, one or more compounds and / or compositions as described herein may be used, for example, to prevent axonal degeneration distal to axonal injury.

[0053] In some embodiments, one or more compounds and / or compositions as described herein may be used, for example, to treat one or more neurodegenerative diseases, disorders, or conditions selected from neuropathy or axonopathy. In some embodiments, one or more compounds and / or compositions as described herein may be used, for example, to treat neuropathy or axonopathy associated with axonal degeneration. In some embodiments, the neuropathy associated with axonal degeneration is a hereditary or congenital neuropathy or axonopathy. In some embodiments, the neuropathy associated with axonal degeneration is caused by a de novo mutation or a somatic mutation. In some embodiments, the neuropathy associated with axonal degeneration is selected from the list included herein. In some implementations, neuropathy or axonopathy is associated with axonal degeneration, including but not limited to Parkinson's disease, Parkinson's syndromes, or Parkinson's plus syndromes, such as multiple system atrophy (MSA), progressive supranuclear palsy (PSP) and corticobasal degeneration, Alzheimer's disease, herpes infection, diabetes, amyotrophic lateral sclerosis (ALS), demyelinating diseases such as multiple sclerosis, ischemia or stroke, chemical injury, thermal injury, and AIDS.

[0054] In some embodiments, the subject administering the compounds or compositions described herein may be or may include a subject who has or is susceptible to a neurodegenerative disease, disorder, or condition. In some embodiments, the neurodegenerative disease, disorder, or condition may be or may include traumatic neuronal injury. In some embodiments, traumatic neuronal injury is blunt force trauma, closed head injury, open head injury, exposure to concussion and / or explosive forces, or penetrating injury to or in or to a region of the brain or its innervation. In some embodiments, traumatic neuronal injury is a force that causes axonal deformation, stretching, crushing, or sheer.

[0055] In some embodiments, the provided method includes administering the compound described herein to a patient in need. In some such embodiments, the patient is at risk of developing a condition characterized by axonal degeneration. In some embodiments, the patient already has a condition characterized by axonal degeneration. In some embodiments, the patient has been diagnosed with a condition characterized by axonal degeneration.

[0056] In some embodiments, the provided method includes applying the composition described herein to a patient population in need. In some embodiments, the population consists of individuals engaged in activities with a high likelihood of traumatic neuronal injury. In some embodiments, the population consists of athletes engaged in contact sports or other high-risk activities.

[0057] In some implementations, the patient is at risk of developing neurodegenerative diseases. In some implementations, the patient is middle-aged or elderly. In some implementations, the patient is known to have genetic risk factors for neurodegeneration.

[0058] In some embodiments, this disclosure provides compounds that can be used, for example, as analytical tools, probes in bioassays, or as therapeutic agents of this disclosure. The compounds provided in this disclosure can also be used to study SARM1 function in biological and pathological phenomena and to evaluate novel SARM1 activity inhibitors in vitro or in vivo.

[0059] In some embodiments, one or more compounds and / or compositions as described herein can be used, for example, as a method to inhibit neuronal degeneration originating from a subject. In some embodiments, one or more compounds and / or compositions as described herein can be used to inhibit degeneration of neurons or portions thereof cultured in vitro. In some embodiments, one or more compounds and / or compositions as described herein can be used as stabilizers to promote neuronal survival in vitro.

[0060] In one embodiment, this disclosure provides a method comprising administering to a subject the compound as described above or a pharmaceutically acceptable salt thereof, the subject (i) having a condition characterized by axonal degeneration; or (ii) being at risk of developing a condition characterized by axonal degeneration.

[0061] In one embodiment, this disclosure provides a method for treating or preventing axonal degeneration, comprising administering to a subject in need of the compound as described above or a pharmaceutically acceptable salt thereof.

[0062] In one embodiment, this disclosure provides a method for inhibiting SARM1, comprising exposing a biological sample to the compound as described above or a pharmaceutically acceptable salt thereof.

[0063] In one embodiment, this disclosure provides a method for treating a patient with amyotrophic lateral sclerosis (ALS), comprising administering to a patient in need of such treatment an effective amount of the compound as described above or a pharmaceutically acceptable salt thereof.

[0064] In one embodiment, this disclosure provides a method for treating a patient with multiple sclerosis, comprising administering to a patient in need of such treatment an effective amount of the compound as described above or a pharmaceutically acceptable salt thereof.

[0065] In one embodiment, this disclosure provides a method for treating a patient with progressive supranuclear palsy, comprising administering to a patient in need of such treatment an effective amount of the compound as described above or a pharmaceutically acceptable salt thereof.

[0066] In one embodiment, this disclosure provides a compound as described above or a pharmaceutically acceptable salt thereof for use in a therapy.

[0067] In one embodiment, this disclosure provides a compound as described above or a pharmaceutically acceptable salt thereof for the treatment of amyotrophic lateral sclerosis (ALS).

[0068] In one embodiment, this disclosure provides the compound as described above or a pharmaceutically acceptable salt thereof for the treatment of multiple sclerosis.

[0069] In one embodiment, this disclosure provides a compound as described above or a pharmaceutically acceptable salt thereof for the treatment of progressive supranuclear palsy. Brief description of the attached diagram

[0071] Figure 1 Example: Structure of the SARM1 protein.

[0072] definition

[0073] Aliphatic group: The term "aliphatic group" refers to a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon (also referred to herein as "carbocyclic" or "cycloaliphatic") that is fully saturated or contains one or more unsaturated units but is not aromatic. Unless otherwise specified, an aliphatic group contains 1-6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-5 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-4 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-3 aliphatic carbon atoms, and in other embodiments, an aliphatic group contains 1-2 aliphatic carbon atoms. In some embodiments, "alicyclic" (or "carbocyclic") refers to a fully saturated or bicyclic C7-C8 hydrocarbon that is fully saturated or contains one or more unsaturated units but is not aromatic. 10Hydrocarbons. Suitable aliphatic groups include, but are not limited to, straight or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl, alkylene, alkenyl, alkynyl and their hybrids.

[0074] Alkyl: The term "alkyl" used alone or as a major component refers to a saturated, optionally substituted straight-chain or branched or cyclic hydrocarbon group having 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2 carbon atoms. The term "cycloalkyl" refers to a saturated ring system with optional substitution of about 3 to about 10 ring carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0075] Alkylene: The term "alkylene" refers to a divalent alkyl group. In some embodiments, "alkylene" is a divalent straight-chain or branched alkyl group. In some embodiments, the "alkylene chain" is polymethylene, i.e., -(CH2). n- - where n is a positive integer, such as 1-6, 1-4, 1-3, 1-2, or 2-3. The optionally substituted alkylene chain is polymethylene, wherein one or more methylene hydrogen atoms are optionally replaced by substituents. Suitable substituents include those described below for substituted aliphatic groups and also those described in this specification. It should be understood that the two substituents of the alkylene chain can together form a ring system. In some embodiments, the two substituents can together form a 3- to 7-membered ring. Substituents can be on the same or different atoms.

[0076] Alkenyl: The term "alkenyl," used alone or as a major component, refers to an optionally substituted straight-chain, branched, or cyclic hydrocarbon group having at least one double bond and having 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3 carbon atoms. The term "cycloalkenyl" refers to an optionally substituted non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond and having about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include cyclopentyl, cyclohexenyl, and cycloheptenyl.

[0077] Alkynyl: The term “alkynyl” used alone or as a major component refers to an optionally substituted straight-chain or branched hydrocarbon group having at least one triple bond and having 2-12, 2-10, 2-8, 2-6, 2-4 or 2-3 carbon atoms.

[0078] Aryl: The term "aryl" refers to a monocyclic or bicyclic system having a total of 5-14 ring members, wherein at least one ring in the system is aromatic, and wherein each ring in the system contains 3-7 ring members. The term "aryl" is used interchangeably with the term "aryl ring." In some embodiments of the invention, "aryl" refers to an aromatic ring system, including but not limited to phenyl, biphenyl, naphthyl, anthracene, etc., which may have one or more substituents. As used herein, the scope of the term "aryl" also includes groups in which the aromatic ring is fused with one or more non-aromatic carbocyclic or heterocyclic rings, such as indanyl, phthalimide, naphthimide, phenanthridine, tetrahydronaphthyl, imidazoalkyl, imidazolidin-2-one, etc.

[0079] Binding: It should be understood that the term “binding” as used herein generally refers to a non-covalent binding between two or more entities. “Direct” binding involves physical contact between entities or parts; indirect binding involves physical interaction through physical contact with one or more intermediate entities. Binding between two or more entities can generally be evaluated in any of a variety of contexts—including in isolation or in the context of more complex systems (e.g., when covalently or otherwise associated with a carrier entity and / or in biological systems or cells) where interacting entities or parts are studied.

[0080] Biological Sample: As used herein, the term "biological sample" typically refers to a sample obtained or derived from a biological source of interest (e.g., tissue or organism or cell culture), as described herein. In some embodiments, the source of interest includes an organism, such as an animal or human. In some embodiments, the biological sample is or contains biological tissue or fluid. In some embodiments, the biological sample may be or contains bone marrow; blood; blood cells; ascites; tissue or fine-needle biopsy samples; body fluids containing cells; free floating nucleic acid; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph; gynecological fluids; skin swabs; vaginal swabs; oral swabs; nasal swabs; washes or lavages, such as catheter lavages or bronchoalveolar lavages; aspirates; scrapings; bone marrow samples; tissue biopsy specimens; surgical specimens; feces, other body fluids, secretions and / or excretions; and / or cells derived therefrom. In some embodiments, the biological sample is or contains cells derived from an individual. In some embodiments, the obtained cells are cells from the individual who obtained the sample or include cells from the individual who obtained the sample. In some embodiments, the sample is a “primary sample” obtained directly from the source of interest by any suitable means. For example, in some embodiments, a primary biological sample is obtained by a method selected from: biopsy (e.g., fine-needle aspiration or tissue biopsy), surgery, collection of bodily fluids (e.g., blood, lymph, feces, etc.). In some embodiments, as will be clear from the context, the term “sample” refers to an article obtained by processing (e.g., by removing one or more components and / or by adding one or more reagents) a primary sample. For example, using semi-permeable membrane filtration. Such a “processed sample” may contain, for example, nucleic acids or proteins extracted from the sample or obtained by subjecting the primary sample to techniques such as amplification or reverse transcription of mRNA, separation and / or purification of certain components.

[0081] Biomarker: The term "biomarker" is used herein to refer to an entity, event, or characteristic whose presence, level, extent, type, and / or form is associated with a particular biological event or state of interest, such that it is considered a "marker" of that event or state. To give only a few examples, in some embodiments, a biomarker may be or comprise a marker of a particular disease state, or a marker of the likelihood that a particular disease, disorder, or symptom may develop, occur, or recur. In some embodiments, a biomarker may be or comprise a marker of a particular disease or treatment outcome or its likelihood. Thus, in some embodiments, a biomarker is predictive of the relevant biological event or state of interest; in some embodiments, a biomarker is prognostic of the relevant biological event or state of interest; and in some embodiments, a biomarker is diagnostic of the relevant biological event or state of interest. A biomarker may be or comprise an entity of any chemical class and may be or comprise a combination of entities. For example, in some embodiments, a biomarker may be or comprise nucleic acids, peptides, lipids, carbohydrates, small molecules, inorganic reagents (e.g., metals or ions), or combinations thereof. In some embodiments, a biomarker is a cell surface marker. In some embodiments, a biomarker is intracellular. In some embodiments, biomarkers are detected extracellularly (e.g., secreted or otherwise produced or present outside cells, such as in bodily fluids like blood, urine, tears, saliva, cerebrospinal fluid, etc.). In some embodiments, biomarkers may be or comprise a genetic or epigenetic signature. In some embodiments, biomarkers may be or comprise a gene expression signature.

[0082] In some embodiments, the biomarker may be or include a marker of neurodegeneration, or a marker of the likelihood that a neurodegenerative disease, disorder, or condition may develop, occur, or recur. In some embodiments, the biomarker may be or include a marker of neurodegeneration, treatment outcome, or its likelihood. Therefore, in some embodiments, the biomarker is predictive of a neurodegenerative disease, disorder, or condition; in some embodiments, the biomarker is prognostic of a neurodegenerative disease, disorder, or condition; and in some embodiments, the biomarker is diagnostic of a neurodegenerative disease, disorder, or condition. In some embodiments, changes in biomarker levels can be detected via cerebrospinal fluid (CSF), plasma, and / or serum.

[0083] In some embodiments, neurodegeneration can be assessed, for example, by detecting increases and / or decreases in the concentrations of neurofilament light chain (NF-L) and / or neurofilament heavy chain (NF-H) (or their phosphorylated form (PNF-H)) in the subject's CSF or blood / plasma. In some embodiments, the incidence and / or progression of neurodegeneration can be assessed using positron emission tomography (PET) with synaptic vesicle glycoprotein 2A (SV2A) ligand. In some embodiments, detectable changes in constitutive NAD and / or cADPR levels in neurons can be used to assess neurodegeneration.

[0084] In some embodiments, detectable changes in one or more neurodegeneration-related proteins in a subject relative to a healthy reference population can be used as biomarkers of neurodegeneration. Such proteins include, but are not limited to, albumin, amyloid-β (Aβ)38, Aβ40, Aβ42, glial fibrillary acidic protein (GFAP), heart-type fatty acid-binding protein (hFABP), monocyte chemoattractant protein (MCP)-1, neurogranulin, neuron-specific enolase (NSE), soluble amyloid precursor protein (sAPP)α, sAPPβ, soluble trigger receptor (sTREM)2 expressed on bone marrow cells, phosphate-tau, and / or total-tau. In some embodiments, increases in cytokines and / or chemokines (including, but not limited to, Ccl2, Ccl7, Ccl12, Csf1, and / or Il66) can be used as biomarkers of neurodegeneration.

[0085] Carrier: As used herein, the term "carrier" refers to a diluent, adjuvant, excipient, or medium applied with the composition. In some exemplary embodiments, the carrier may include sterile liquids, such as water and oils, including petroleum, animal, plant, or synthetic oils, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. In some embodiments, the carrier is or comprises one or more solid components.

[0086] Combination therapy: As used herein, the term "combination therapy" refers to those situations in which a subject is simultaneously exposed to two or more treatment regimens (e.g., two or more therapeutic agents). In some embodiments, two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all "dose" of the first regimen are administered before any dose of the second regimen); in some embodiments, such agents are administered in an overlapping dosing regimen. In some embodiments, "administration" of combination therapy may involve administering one or more agents or methods to a subject receiving other agents or methods in the combination. For clarity, combination therapy does not require that the individual agents be administered together in a single composition (or even simultaneously), however, in some embodiments, two or more agents or their active portions may be administered together in a combination composition or even in a combination compound (e.g., as part of a single chemical complex or covalent entity).

[0087] Composition: Those skilled in the art will understand that the term "composition" can be used to refer to a discrete physical entity comprising one or more specified components. Generally, unless otherwise stated, a composition can be in any form, such as a gas, gel, liquid, solid, etc.

[0088] Domain: As used herein, the term "domain" refers to a segment or portion of an entity. In some embodiments, a "domain" is associated with a specific structural and / or functional feature of an entity such that it substantially or completely retains that specific structural and / or functional feature when physically separated from the remainder of its parent entity. Alternatively or additionally, a domain may be or comprise a portion of an entity that, when separated from the (parental) entity and linked to a different (recipient) entity, substantially retains and / or imparts one or more of its structural and / or functional features, which are characteristic of the parent entity, to the recipient entity. In some embodiments, a domain is a segment or portion of a molecule (e.g., a small molecule, carbohydrate, lipid, nucleic acid, or polypeptide). In some embodiments, a domain is a portion of a polypeptide; in some such embodiments, the domain is characterized by specific structural elements (e.g., specific amino acid sequences or sequence motifs, α-helix features, β-sheet features, coil-coil features, random coil features, etc.) and / or specific functional features (e.g., binding activity, enzymatic activity, folding activity, signal transduction activity, etc.).

[0089] Dosage Form or Unit Dosage Form: Those skilled in the art will understand that the term "dosage form" can be used to refer to a physically discrete unit of an active agent (e.g., a therapeutic or diagnostic agent) intended for administration to a subject. Typically, each such unit contains a predetermined amount of the active agent. In some embodiments, such an amount is a unit dose (or a portion thereof) suitable for administration according to a dosing regimen that has been determined to be associated with desired or beneficial outcomes when administered to the relevant population (i.e., a therapeutic dosing regimen). Those skilled in the art will understand that the total amount of a therapeutic composition or agent administered to a particular subject is determined by one or more attending physicians and may involve the administration of multiple dosage forms.

[0090] Dosing regimen or treatment regimen: Those skilled in the art will understand that the terms "dosing regimen" and "treatment regimen" can be used to refer to a set of unit doses (usually more than one) that are typically administered individually to a subject over time periods. In some embodiments, a given therapeutic agent has a recommended dosing regimen that may involve one or more doses. In some embodiments, a dosing regimen includes multiple doses, each time separate from the other doses. In some embodiments, the individual doses are separated from each other by the same length of time period; in some embodiments, a dosing regimen includes multiple doses and at least two distinct time periods separating the individual doses. In some embodiments, all doses within a dosing regimen have the same unit dose. In some embodiments, the different doses within a dosing regimen have different amounts. In some embodiments, a dosing regimen includes a first dose of a first administered amount, followed by one or more additional doses of a second administered amount different from the first administered amount. In some embodiments, a dosing regimen includes a first dose of a first administered amount, followed by one or more additional doses of a second administered amount identical to the first administered amount. In some embodiments, when administered in a relevant population, the dosing regimen is associated with a desired or beneficial outcome (i.e., a therapeutic dosing regimen).

[0091] Excipients: As used herein, refers to non-therapeutic agents that may be included in a pharmaceutical composition, for example, to provide or contribute to a desired consistency or stabilizing effect. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerin, propylene, diols, water, ethanol, etc.

[0092] Heteroaryl: The terms "heteroaryl" and "heteroaryl-" used alone or as part of a larger group such as "heteroarylalkyl" or "heteroarylalkoxy" refer to a group having 5-10 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6, 10, or 14 π-electrons shared in a cyclic array; and having one to five heteroatoms in addition to a carbon atom. The term "heteroatom" refers to nitrogen, oxygen, or sulfur and includes any oxidized form of nitrogen or sulfur, as well as any quaternized form of basic nitrogen. Heteroaryl includes, but is not limited to, thiophene, furanyl, pyrrole, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyrazinyl, indazinyl, purinyl, naphridinyl, and pteridinyl. As used herein, the terms “heteroaryl” and “heteroary-” also include groups in which a heteroaryl ring is fused with one or more aryl, alicyclic, or heterocyclic rings. Non-limiting examples include indolyl, isoindolyl, benzothiopheneyl, benzofuranyl, dibenzofuranyl, indazoleyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, cenolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinazinyl, carbazolyl, acridineyl, phenazinyl, phenthiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic or bicyclic. The term “heteroaryl” is used interchangeably with the terms “heteroaryl ring,” “heteroaryl group,” or “heteroaryl group,” any of which includes an optionally substituted ring. The term "heteroaryl" refers to an alkyl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl portions are optionally substituted independently.

[0093] Heterocycle: As used herein, the terms “heterocycle,” “heterocyclic group,” “heterocyclic residue,” and “heterocycle” are used interchangeably and refer to a specified 3- to 8-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is saturated or partially unsaturated and has one or more, for example, one to four, heteroatoms as defined above, in addition to a carbon atom. When referring to the ring atom of a heterocycle, the term “nitrogen” includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur, and nitrogen, nitrogen can be N (as in 3,4-dihydro-2H-pyrrole), NH (as in pyrrolidinyl), or NR. + (e.g., in N-substituted pyrrolidinyl groups). The heterocycle can be attached to its side group at any heteroatom or carbon atom that produces a stable structure, and any ring atom can optionally be substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, decahydroquinolinyl, oxazolyl, piperazine, dioxalyl, dioxopentyl, and diazapyridine. Basic, oxygen and nitrogen Basic, sulfur-nitrogen The heterocyclic group can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, and more preferably monocyclic or bicyclic. The term "heterocyclic alkyl" refers to an alkyl group substituted with a heterocyclic group, wherein the alkyl and heterocyclic portions are optionally substituted independently. Additionally, heterocycles also include groups in which the heterocycle is fused to one or more aryl rings (e.g., 2,3-dihydrobenzofuran, 2,3-dihydrobenzo[b][1,4]dioxin, etc.).

[0094] Hydrogen: As used in this article, the term "hydrogen" is used to refer to all three isotopes of hydrogen, namely protium (…). 1 H), deuterium ( 2 H) and tritium ( 3 H).

[0095] Inhibitor: As used herein, the term "inhibitor" refers to an entity, condition, or event whose presence, level, or extent is associated with a reduction in the level or activity of a target. In some embodiments, an inhibitor may act directly (in which case it directly exerts its effect on its target, e.g., by binding to the target); in some embodiments, an inhibitor may act indirectly (in which case it exerts its effect by interacting with a modulator of the target and / or otherwise altering a modulator of the target, resulting in a reduction in the level and / or activity of the target). In some embodiments, an inhibitor is an inhibitor whose presence or level is associated with a reduced target level or activity relative to a specific reference level or activity (e.g., observed under appropriate reference conditions, such as the presence of a known inhibitor or the absence of the inhibitor in question, etc.).

[0096] Neurodegeneration: As used herein, the term "neurodegeneration" refers to a reduction in one or more features, structures, functions, or properties of neurons or neuronal tissue. In some embodiments, neurodegeneration is observed as a pathological reduction in an organism. Those skilled in the art will understand that neurodegeneration is associated with certain diseases, conditions, and illnesses, including those affecting humans. In some embodiments, neurodegeneration can be transient (e.g., sometimes associated with certain infections and / or chemical or mechanical damage); in some embodiments, neurodegeneration can be chronic and / or progressive (e.g., generally associated with certain diseases, disorders, or illnesses, such as, but not limited to, Parkinson's disease, amyotrophic lateral sclerosis, multiple sclerosis, Huntington's disease, or Alzheimer's disease). In some embodiments, neurodegeneration can be assessed, for example, by detecting an increase in a neurodegeneration-related biomarker in a subject. In some embodiments, neurodegeneration can be assessed, for example, by detecting a decrease in a neurodegeneration-related biomarker in a subject. Optionally or additionally, in some embodiments, neurodegeneration can be assessed by magnetic resonance imaging (MRI), CSF, or biomarkers contained in blood / plasma, or other biomarkers observed in the patient. In some embodiments, neurodegeneration is defined as a score below 24 on a mini-mental state examination. In some embodiments, neurodegeneration refers to the loss of synapses. In some embodiments, neurodegeneration refers to a reduction in neural tissue associated with traumatic injury (e.g., exposure to external forces that disrupt the integrity of neural tissue). In some embodiments, neurodegeneration refers to a reduction in peripheral neural tissue. In some embodiments, neurodegeneration refers to a reduction in central nervous tissue.

[0097] Oral administration: As used herein, the terms “oral administration” and “by oral administration” have the meaning as understood in the art and refer to the administration of a compound or composition through the oral cavity.

[0098] Parenteral administration: As used herein, the terms “parenteral administration” and “by means of administration” have their meaning as understood in the art, referring to administration other than enteric and local administration, usually by injection, including but not limited to intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injections and infusions.

[0099] Partially unsaturated: As used herein, the term “partially unsaturated” refers to a ring moiety comprising at least one double or triple bond between ring atoms. The term “partially unsaturated” is intended to include rings having multiple unsaturated sites, but not necessarily aromatic moieties (e.g., aryl or heteroaryl) as defined herein.

[0100] Patient: As used herein, the term "patient" means any organism to which the provided composition is applied, or may be applied, for example, experimental, diagnostic, preventative, cosmetic, and / or therapeutic purposes. Typical patients include animals (e.g., mammals, such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, the patient is a human. In some embodiments, the patient has or is susceptible to one or more diseases or conditions. In some embodiments, the patient exhibits one or more symptoms of a disease or condition. In some embodiments, the patient has been diagnosed with one or more diseases or conditions. In some embodiments, the patient is receiving or has received certain treatments for the diagnosis and / or treatment of a disease, disorder, or condition.

[0101] Pharmaceutical Composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose suitable for administration in a treatment or dosing regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition may be specifically formulated for administration in solid or liquid form, including those suitable for: oral administration, such as infusions (aqueous or non-aqueous solutions or suspensions), tablets, such as those for oral, sublingual, and systemic absorption, pills, powders, granules, pastes for application to the tongue; parenteral administration, such as by subcutaneous, intramuscular, intravenous, or epidural injection, as, for example, a sterile solution or suspension or a sustained-release formulation; topical administration, such as as a cream, ointment, controlled-release patch, or spray for application to the skin, lungs, or mouth; intravaginal or rectal administration, such as as a vaginal suppository, cream, or foam; sublingual; ocular; transdermal; or nasal, pulmonary, and other mucosal surfaces.

[0102] Pharmaceutically acceptable: As used herein, the term “pharmaceutically acceptable” means, to a reasonable extent of medical judgment, those compounds, materials, compositions and / or dosage forms that are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications, and in proportion to a reasonable benefit / risk ratio.

[0103] Pharmaceutically acceptable carriers: As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, which participates in carrying or transporting the subject compound from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of compatibility with other components of the formulation and harmlessness to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth gum; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffers, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic and compatible substances used in pharmaceutical formulations.

[0104] Pharmaceutically acceptable salts: As used herein, the term "pharmaceutically acceptable salt" means a salt of such compounds suitable for use in a pharmaceutical context, i.e., a salt suitable for contact with tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., within the bounds of reasonable medical judgment, and in proportion to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SMBerge et al. described pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66:1-19 (1977). In some embodiments, pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts, which are salts formed by reacting an amino group with an inorganic acid (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or an organic acid (e.g., acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid) or by using other methods used in the art (e.g., ion exchange). In some implementations, pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, disaccharide, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-heptahydrate, glyceryl phosphate, gluconate, hemisulfate, heptahydrate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. In some embodiments, pharmaceutically acceptable salts may, where appropriate, include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions, such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl, sulfonate, and arylsulfonate ions having 1 to 6 carbon atoms.

[0105] Prevention or avoidance: As used herein, the terms “prevention” or “avoidance” when used in connection with the occurrence of a disease, disorder, or condition refer to reducing the risk of developing a disease, disorder, or condition and / or delaying the onset of one or more features or symptoms of a disease, disorder, or condition. Prevention is considered complete when the onset of a disease, disorder, or condition has been delayed for a predefined period of time.

[0106] Specificity: When referring herein to an active reagent, the term "specificity" is understood by those skilled in the art to mean that the reagent distinguishes a potential target entity or state. For example, in some embodiments, a reagent is said to bind "specifically" to its target if it preferentially binds to that target in the presence of one or more competing alternative targets. In many embodiments, specific interactions depend on the presence of specific structural features of the target entity (e.g., epitopes, fissures, binding sites). It should be understood that specificity does not need to be absolute. In some embodiments, specificity can be evaluated relative to the specificity of the binder to one or more other potential target entities (e.g., competitors). In some embodiments, specificity is evaluated relative to the specificity of a reference specific binder. In some embodiments, specificity is evaluated relative to the specificity of a reference non-specific binder. In some embodiments, the reagent or entity binds to a competing alternative target undetectably when bound to its target entity. In some embodiments, the binder binds to its target entity with a higher binding rate, a lower dissociation rate, increased affinity, reduced dissociation, and / or increased stability compared to a competing alternative target.

[0107] Subject: As used herein, the term "subject" refers to an organism, typically a mammal (e.g., a human, including prenatal human forms in some embodiments). In some embodiments, the subject suffers from a relevant disease, disorder, or condition. In some embodiments, the subject is susceptible to a disease, disorder, or condition. In some embodiments, the subject exhibits one or more symptoms or characteristics of a disease, disorder, or condition. In some embodiments, the subject does not exhibit any symptoms or characteristics of a disease, disorder, or condition. In some embodiments, the subject is a person with one or more characteristics of susceptibility or risk to a disease, disorder, or condition. In some embodiments, the subject is a patient. In some embodiments, the subject is an individual who has received and / or has received a diagnostic and / or therapeutic treatment.

[0108] Substituted or Optionally Substituted: As described herein, compounds of the present invention may comprise an “optionally substituted” portion. Generally, the term “substituted,” regardless of whether it is preceded by the term “optionally,” means that one or more hydrogens of the specified portion are replaced by suitable substituents. “Substituted” applies to one or more hydrogens explicitly or implicitly present in the structure (e.g., It means at least and It means at least Unless otherwise indicated, "optionally substituted" groups may have stable substituents at their respective substituted positions, and when more than one position in any specified structure can be substituted by more than one substituent selected from the specified group, the substituents may be the same or different at each position. The combinations of substituents contemplated in this invention are preferably those that result in the formation of stable or chemically viable compounds. As used herein, the term "stable" means a compound that remains substantially unchanged when subjected to conditions permissible for its production, testing, and, in some embodiments, its recovery, purification, and use for one or more purposes disclosed herein.

[0109] The suitable monovalent substituent on the substituted carbon atom of the "optionally substituted" group is independently a halogen; –(CH2) 0–4 R°;–(CH2) 0–4 OR°;-O(CH2) 0-4 R°、–O–(CH2) 0–4 C(O)OR°;–(CH2) 0–4 CH(OR°)2;–(CH2) 0– 4SR°; –(CH2) 0–4 Ph, which can be replaced by R°; –(CH2) 0–4 O(CH2) 0–1 Ph, which can be substituted by R°; –CH=CHPh, which can be substituted by R°; –(CH2) 0–4 O(CH2) 0–1 -pyridyl group, which can be substituted by R°; –NO2; –CN; –N3; -(CH2) 0–4 N(R°)2;–(CH2) 0–4 N(R°)C(O)R°; –N(R°)C(S)R°; –(CH2) 0–4 N(R°)C(O)NR°2; -N(R°)C(S)NR°2; –(CH2) 0–4 N(R°)C(O)OR°; –N(R°)N(R°)C(O)R°; –N(R°)N(R°)C(O)NR°2; –N(R°)N(R°)C(O)OR°; –(CH2) 0–4 C(O)R°; –C(S)R°; –(CH2) 0–4 C(O)OR°;–(CH2) 0–4 C(O)SR°;-(CH2) 0–4 C(O)OSiR°3;–(CH2) 0–4 OC(O)R°;–OC(O)(CH2) 0–4 SR°;–(CH2) 0–4 SC(O)R°;–(CH2)0–4 C(O)NR°2; –C(S)NR°2; –C(S)SR°; –SC(S)SR°, -(CH2) 0–4 OC(O)NR°2; -C(O)N(OR°)R°; –C(O)C(O)R°; –C(O)CH2C(O)R°; –C(NOR°)R°; –(CH2) 0–4 SSR°;–(CH2) 0–4 S(O)2R°;–(CH2) 0–4 S(O)(NH)R°;–(CH2) 0–4 S(O)₂OR°;–(CH₂) 0–4 OS(O)2R°; –S(O)2NR°2; -(CH2) 0–4 S(O)R°; -N(R°)S(O)2NR°2; –N(R°)S(O)2R°; –N(OR°)R°; –C(NH)NR°2; –P(O)2R°; –P(O)R°2; –OP(O)R°2; –OP(O)(OR°)2; SiR°3; –(C 1–4 (linear or branched alkylene)O–N(R°)2; or –(C 1–4 (straight-chain or branched alkylene)C(O)O–N(R°)2, wherein each of R° can be substituted as defined below and independently be hydrogen, C 1–6 Aliphatic groups, –CH2Ph, –O(CH2) 0–1 Ph, -CH2- (5- to 6-membered heteroaryl ring), 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, or 8- to 10-membered bicyclic aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, or, although as defined above, two independently occurring R° together with their intervening atom form a 3- to 12-membered saturated, partially unsaturated or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, which may be substituted as defined below.

[0110] Suitable monovalent substituents on R° (or the ring formed by two independently occurring R° and their intervening atoms) are independently halogens, –(CH2). 0–2 R · –(halogenated R) · ), –(CH2) 0–2 OH, –(CH2) 0–2 OR · –(CH2) 0–2 CH(OR · )2;-O(halogenated R · –CN, –N3, –(CH2) 0–2C(O)R · –(CH2) 0–2 C(O)OH, –(CH2) 0–2 C(O)OR · –(CH2) 0–2 SR · –(CH2) 0– 2SH、–(CH2) 0–2 NH2、–(CH2) 0–2 NHR · –(CH2) 0–2 NR · 2. –NO2, –SiR · 3. –OSiR · 3. -C(O)SR · , – (C 1–4 (straight-chain or branched alkylene)C(O)OR · Or –SSR · , where R · Each is either not replaced or, if "halogenated" comes first, is replaced by only one or more halogens, and is independently selected from C. 1–4 Aliphatic groups, –CH2Ph, –O(CH2) 0–1 Ph or a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Suitable divalent substituents on the saturated carbon atom of R° include =O and =S.

[0111] Suitable divalent substituents on the saturated carbon atom of the "optionally substituted" group include the following: =O ("oxo"), =S, =NNR. * 2、=NNHC(O)R * =NNHC(O)OR * =NNHS(O)2R * =NR * =NOR * 、 –O(C(R) * 2)) 2–3 O – or –S(C(R) * 2)) 2–3 S–, where each independently occurring R * Selected from hydrogen, C can be substituted as defined below 1–6 An aliphatic group, or an unsubstituted, 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 independent heteroatoms selected from nitrogen, oxygen, and sulfur. Suitable divalent substituents for the ortho-substituted carbon of the "optionally substituted" group include: –O(CR * 2) 2–3 O–, where each independently occurring R* Selected from hydrogen, C can be substituted as defined below 1–6 Aliphatic group, or unsubstituted 5–6–membered saturated, partially unsaturated or aryl ring having 0–4 heteroatoms independently selected from nitrogen, oxygen and sulfur.

[0112] R * Suitable substituents on aliphatic groups include halogens, -R · -(halogenated R) · -OH, -OR · –O (halogenated R) · ), –CN, –C(O)OH, –C(O)OR · –NH2, –NHR · –NR · 2 or –NO2, where R · Each is either not replaced, or if "halogenation" precedes it, it is replaced by only one or more halogens, and is independently C. 1–4 Aliphatic groups, –CH2Ph, –O(CH2) 0–1 Ph or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0113] Suitable substituents on the substituted nitrogen of the "optionally substituted" group include or in Each of the following is an independent C that is hydrogen and can be substituted as defined below. 1–6 Aliphatic groups, unsubstituted –OPh, or unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl rings having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or, despite the above definitions, the presence of two independent amounts Together with its intercalary atoms, it forms an unsubstituted 3- to 12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring with 0 to 4 independent heteroatoms selected from nitrogen, oxygen, and sulfur.

[0114] Suitable substituents on the aliphatic group can be halogens, -R · -(halogenated R) · –OH, –OR · –O (halogenated R) · ), –CN, –C(O)OH, –C(O)OR · –NH2, –NHR · –NR · 2 or -NO2, where R ·Each is either not replaced, or if "halogenation" precedes it, it is replaced by only one or more halogens, and is independently C. 1–4 Aliphatic groups, –CH2Ph, –O(CH2) 0–1 Ph or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0115] Therapeutic agent: As used herein, the term "therapeutic agent" generally refers to any agent that, when administered to an organism, induces the desired pharmacological effect. In some embodiments, an agent is considered a therapeutic agent if it exhibits a statistically significant effect in an appropriate population. In some embodiments, an appropriate population may be a population of model organisms. In some embodiments, an appropriate population may be defined by various criteria, such as an age group, sex, genetic background, pre-existing clinical condition, etc. In some embodiments, a therapeutic agent is a substance that can be used to alleviate, improve, relieve, suppress, prevent, delay the onset of one or more symptoms or features of a disease, disorder, or condition, reduce its severity, and / or reduce its incidence. In some embodiments, a "therapeutic agent" is an agent that has been or requires approval by a government agency before it can be marketed for administration to humans. In some embodiments, a "therapeutic agent" is an agent that requires a medical prescription for administration to humans.

[0116] Treatment: As used herein, the terms “treat,” “treatment,” or “treating” refer to any method used to partially or completely relieve, improve, alleviate, suppress, prevent, delay the onset, reduce the severity, and / or decrease the incidence of one or more symptoms or features of a disease, disorder, or condition. Treatment may be administered to subjects who do not exhibit signs of a disease, disorder, or condition. In some implementations, treatment may be administered to subjects who exhibit only early signs of a disease, disorder, or condition, for example, for the purpose of reducing the risk of developing a pathological condition associated with the disease, disorder, or condition.

[0117] Detailed description of certain implementation schemes

[0118] Programmed axial mutation and SARM1

[0119] Axonal degeneration is a major pathological feature of neurological diseases, including but not limited to Alzheimer's disease, Parkinson's disease, ALS, multiple sclerosis, diabetic peripheral neuropathy, chemotherapy-induced peripheral neuropathy, hereditary neuropathy, traumatic brain injury, and / or glaucoma. Damaged or unhealthy axons are eliminated through an inherent self-destruction program that differs from conventional cell death pathways such as apoptosis known as Wallerian degeneration (Gerdts, J. et al., Neuron, 2016, 89, 449-460; Whitmore, AV et al., Cell Death Differ., 2003, 10, 260-261). In Wallerian degeneration, peripheral nerves undergo selective destruction of axonal segments distal to the injury, while proximal axonal segments and cell bodies remain intact. This type of denaturation is characterized by, first, the depletion of nicotinamide mononucleotide adenosine transferase (NMNAT), followed by the loss of nicotinamide adenine dinucleotide (NAD+), adenosine triphosphate (ATP) loss, neurofilament protein hydrolysis, and finally, axonal degradation approximately 8 to 24 hours after injury (Gerdts, J., et al., Neuron, 2016, 89, 449-460).

[0120] NAD+ is a ubiquitous metabolite that plays a crucial role in energy metabolism and cell signaling (Belenkey et al., Trends Biochem., 2007, 32, 12-19; ​​Chiarugi et al., Nat. Rev. Cancer, 2012, 12, 741-752). Homeostatic regulation of NAD+ levels is also responsible for maintaining axonal stability and integrity. Therefore, manipulations that increase NMNAT1 axonal localization confer axonal protection (Babetto et al., Cell Rep., 2010, 3, 1422-1429; Sasaki et al., J. Neurosci., 2009).

[0121] In genome-wide RNAi screening of primary mouse neurons, a sterile α- and TIR motif 1 (SARM1) was identified, with SARM1 knockout resulting in durable protection of sensory neurons against damage-induced axonal degeneration (Gerdts et al., J Neurosci, 2013, 33, 13569-13580). SARM1 belongs to the cytoplasmic adaptor protein family but is unique among its members as it is the most evolved and ancient adaptor, anomalously inhibiting TLR signaling and has been identified as a central executor of the damage-induced axonal death pathway (O'Neill, LA & Bowie, AG, Nat. Rev. Immunol., 2007, 7, 353-364; Osterloh, JM et al., Science, 2012, 337, 481-484; Gerdts, J. et al., J. Neurosci. 33, 2013, 13569-13580). SARM1 activation via axonal injury or forced dimerization of the SARM1-TIR domain promotes rapid and catastrophic depletion of nicotinamide adenine dinucleotide (NAD+), followed shortly by axonal degradation, thus highlighting the central role of NAD+ homeostasis in axonal integrity (Gerdts, J. et al., Science, 2015, 348, 453-457). SARM1 is essential for this injury-induced NAD+ depletion in vitro and in vivo, and SARM1 activation locally triggers axonal degeneration through NAD(+) disruption (Gerdts et al., Science, 2015, 348, 452-457; Sasaki et al., J. Biol. Chem. 2015, 290, 17228-17238; both references are incorporated herein by reference in their entirety).

[0122] Studies of genetic loss of function clearly demonstrate that SARML acts as a central executor of the post-traumatic axonal degeneration pathway. SARM1 gene knockout allows axonal preservation for 14 days or longer after nerve transection (Osterloh, JM et al., Science, 2012, 337, 481-484; Gerdts, J. et al., J. Neurosci., 2013, 33, 13569-13580) and also improves functional outcomes in mice after traumatic brain injury (Henninger, N. et al., Brain 139, 2016, 1094-1105). In addition to its role in direct axonal injury, SARM1 is also essential for the axonal degeneration observed in chemotherapy-induced peripheral neuropathy. Loss of SARM1 blocks chemotherapy-induced peripheral neuropathy, inhibits axonal degeneration, and enhances pain sensitivity developed after chemotherapeutic vincristine treatment (Geisler et al., Brain, 2016, 139, 3092-3108).

[0123] SARM1 contains several conserved motifs that mediate oligomerization and protein-protein interactions, including the SAM domain, the ARM / HEAT motif, and the TIR domain. Figure 1 (O'Neill, LA & Bowie, AG, Nat. Rev. Immunol., 2007, 7, 353-364; Tewari, R. et al., Trends Cell Biol., 2010, 20, 470-481; Qiao, F. & Bowie, JU, Sci. STKE 2005, Re7, 2005). TIR domains are commonly found in signaling proteins that function in the innate immune pathway, where they serve as scaffolds for protein complexes (O'Neill, LA & Bowie, AG, Nat. Rev. Immunol., 2007, 7, 353-364). Significantly, dimerization of the SARM1-TIR domain is sufficient to induce axonal degeneration and rapidly trigger NAD+ degradation by acting as an NAD+ cleaving enzyme (Milbrandt et al., WO 2018 / 057989; Gerdts, J. et al., Science, 2015, 348, 453-457). Given the central role of SARM1 in the axonal degeneration pathway and its identified NAD enzyme activity, efforts have been made to identify agents that can modulate SARM1 and potentially act as useful therapeutic agents, for example, to protect against neurodegenerative diseases, including peripheral neuropathy, traumatic brain injury, and / or neurodegenerative disorders.

[0124] This disclosure provides certain compounds and / or compositions as SARM1 inhibitors (e.g., as SARM1 inhibitors), and related technologies.

[0125] compound

[0126] In some embodiments, this disclosure provides compounds of formula I:

[0127]

[0128] Or its pharmaceutically acceptable salt, wherein:

[0129] Ring A is a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur;

[0130] R 1 The substituted group is selected from 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, or 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0131] R x Each group is selected from halogen, cyano, OR, SR, N(R)2, or optionally substituted groups, and is selected from C. 1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbon rings and 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl, and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur;

[0132] Ring B is a saturated 5- to 7-membered heterocycle with a structure And optionally includes another option selected from –NH-, -O- and –NR. 2 - group;

[0133] R is each independently hydrogen, or an optionally substituted group selected from C. 1-6 Aliphatic group, 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur, phenyl, and 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur; or:

[0134] The two R groups together with the nitrogen atom to which they are attached form a 3- to 7-membered monocyclic heterocycle with 0-2 additional heteroatoms independently selected from oxygen, nitrogen and sulfur;

[0135] R 2 Each independently represents a halogen, N(R)2, OR, or C. 1-3 Aliphatic groups or –(C1-3 (Aliphatic group)R 3 ;

[0136] R 3 Each of the groups is an optional substituted group, selected from C. 1-6 Aliphatic group, 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl, 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, 8- to 10-membered bicyclic saturated, partially unsaturated or aryl carbon ring, 8- to 10-membered bicyclic saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, or 8- to 10-membered bicyclic heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur;

[0137] m is 0, 1, or 2; and

[0138] n is 0, 1, or 2.

[0139] Generally, as defined above, ring A is a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, ring A is a 5-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, ring A is pyrroleyl, furanyl, or thiopheneyl. In some embodiments, ring A is a 5-membered heteroaryl ring having 2 to 3 heteroaryl atoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, ring A is a 5-membered heteroaryl ring having 2 heteroaryl atoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, ring A is a group selected from pyrazolyl, imidazolyl, isothiazolyl, and thiazolyl.

[0140] In some embodiments, ring A is a 5-membered heteroaryl ring having three heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some such embodiments, ring A is a group selected from triazolyl and thiadiazolyl.

[0141] In some embodiments, ring A is a 6-membered heteroaryl ring having 1-2 nitrogen atoms. In some embodiments, ring A is pyridinyl. In some embodiments, ring A is pyridin-2(1H)-keto.

[0142] In some implementations, ring A is selected from...

[0143]

[0144]

[0145] In some implementations, ring A is selected from...

[0146]

[0147] Where R x Each of the groups is an optional substituted group, selected from C. 1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbon rings, 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl groups, and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0148] In some implementations, ring A is selected from...

[0149]

[0150] In some implementations, ring A is selected from...

[0151]

[0152] in:

[0153] R on nitrogen atom x Each group is independently selected from the optionally substituted groups, which are selected from C 1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbocyclic rings, 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur, phenyl groups, and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur; and

[0154] R on carbon atom x Each group is independently selected from halogen, cyano, OR, SR, N(R)2, or optionally substituted groups, and is selected from C. 1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbon rings, 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl groups, and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0155] In some implementations, ring A is selected from...

[0156] In some particularly preferred embodiments, ring A is selected from...

[0157] Generally as defined above, R 1 The substituted group is selected from 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, or 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0158] In some implementation schemes, R 1It is a 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 1 It is a 5- to 6-membered saturated or partially unsaturated heterocycle having 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 1 The substituted group is selected from pyrrolidinyl, piperidinyl, morpholinyl, and piperazineyl.

[0159] In some implementation schemes, R 1 It is a 5- to 6-membered heteroaryl ring with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen and sulfur.

[0160] In some implementation schemes, R 1 It is a 5-membered heteroaryl ring with 1-3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 1 It is a 5-membered heteroaryl ring with 1-2 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 1 The substituted group is selected from pyrazolyl, thiazolyl, and thiophene rings.

[0161] In some implementation schemes, R 1 It is an optionally substituted 6-membered heteroaryl ring having 1-3 nitrogen atoms. In some embodiments, R 1 It is an optionally substituted 6-membered heteroaryl ring having 1-2 nitrogen atoms. In some embodiments, R 1 The substituted group is selected from pyridinyl, pyrimidinyl, and pyridazinyl.

[0162] In some implementation schemes, R 1 Selected from

[0163]

[0164] In some particularly preferred embodiments, R 1 Selected from

[0165] Generally as defined above, R x Each group is independently selected from halogen, cyano, OR, SR, N(R)2, or optionally substituted groups, and is selected from C. 1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbocyclic rings, 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur, phenyl groups, and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, R xIt is a halogen. In some such implementations, R x It is chlorine or bromine.

[0166] In some implementation schemes, R x It is a cyano group.

[0167] In some implementation schemes, R x For OR. In some implementations, R x OR, where R is selected from hydrogen and optionally substituted C. 1-6 Aliphatic group. In some embodiments, R x OR, where R is selected from hydrogen and optionally substituted C. 1-4 Aliphatic group. In some embodiments, R x Selected from OH, OCH3 and OCH2CH3.

[0168] In some implementation schemes, R x For SR. In some implementations, R x SR, where R is selected from hydrogen and optionally substituted C. 1-6 Aliphatic group. In some embodiments, R x SR, where R is selected from hydrogen and optionally substituted C. 1-4 Aliphatic group. In some embodiments, R x Selected from SH, SCH3 and SCH2CH3.

[0169] In some implementation schemes, R x For N(R)². In some implementations, R x The form is N(R)₂, where R is selected from hydrogen and optionally substituted C. 1-6 Aliphatic group. In some embodiments, R x The form is N(R)₂, where R is selected from hydrogen and optionally substituted C. 1-4 Aliphatic group. In some embodiments, R x Selected from NH2, NHCH3, NHCH2CH3, N(CH3)2 and N(CH2CH3)2.

[0170] In some implementation schemes, R x C is an optional replacement 1-4 Aliphatic group. In some embodiments, R x C is an optional replacement 3-4 Aliphatic groups. In some such embodiments, R x Selected from tert-butyl,

[0171] In some implementation schemes, R x C 1-4Aliphatic groups, optionally substituted with groups selected from: halogens, –(CH2). 0–4 R°、–(CH2) 0–4 OR°、-(CH2) 0–4 N(R°)2、–(CH2) 0–4 C(O)OR° and –(CH2) 0–4 C(O)NR°2. In some such embodiments, R° is selected from hydrogen, C 1–6 Aliphatic groups, –CH2Ph, –O(CH2) 0–1 Ph, -CH2- (5- to 6-membered heteroaryl ring), 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or 8- to 10-membered bicyclic aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or: two independently occurring R° together with their intermediary atoms to form 3- to 12-membered saturated, partially unsaturated or aryl mono- or bicyclic rings independently selected from nitrogen, oxygen or sulfur.

[0172] In some implementation schemes, R x C 1-4 The aliphatic group is optionally substituted with a group selected from: halogen, –R°, –OR°, -N(R°)2, –C(O)OR°, and –C(O)NR°2. In some embodiments, R… x C that is optionally substituted with halogen 1-4 Aliphatic groups. In some such embodiments, R x Selected from –CH3, –CF3, -CHF2 and CH2F.

[0173] In some implementation schemes, R x Selected from –CH2R°, –CH2OR°, –CH2N(R°)2, –CH2C(O)OR°, and –CH2C(O)N(R°)2. In some such embodiments, R x Selected from –CH2OH, -CH2OCH3, -CH2C(O)NH2, -CH2C(O)NHCH3 and -CH2C(O)N(CH3)2.

[0174] In some implementation schemes, R x The substituted 3- to 7-membered saturated or partially unsaturated carbon rings are used. In some embodiments, R x The substituted 5- to 7-membered saturated or partially unsaturated carbon rings are used. In some embodiments, R x The substituted 5- to 7-membered saturated carbon ring is used. In some such embodiments, R x Selected from substituted cyclopentyl or cyclohexyl groups.

[0175] In some implementation schemes, R x It is a 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R x It is a 3- to 4-membered saturated heterocycle with one optionally substituted heteroatom selected from oxygen, nitrogen, and sulfur. In some embodiments, R x It is a 5- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R x It is a 5- to 7-membered saturated heterocycle with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some such embodiments, R x It is selected from optionally substituted pyrrolidinyl, piperidinyl, piperazineyl and morpholinyl.

[0176] In some implementation schemes, R x The phenyl group is optionally substituted.

[0177] In some implementation schemes, R x It is a 5- to 6-membered heteroaryl ring with 1 to 3 optionally substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R x It is a 5-membered heteroaryl ring with 1-3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R x It is a 5-membered heteroaryl ring with 1-2 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some such embodiments, R x It is selected from pyrrole, pyrazol, imidazole, oxazol, and thiazolyl groups with optional substitution.

[0178] In some implementation schemes, R x It is an optionally substituted 6-membered heteroaryl ring having 1-3 nitrogen atoms. In some embodiments, R x It is an optionally substituted 6-membered heteroaryl ring having 1-2 nitrogen atoms. In some such embodiments, R x Selected from optionally substituted pyridyl, pyridinyl, pyrimidinyl and pyrazinyl groups.

[0179] Generally, as defined above, ring B is a saturated 5- to 7-membered heterocycle with a structure And optionally includes another option selected from –NH-, -O- and –NR. 2 -A group. In some embodiments, ring B is a saturated 5- to 7-membered heterocycle with a structure Ring B also contains another element selected from –NH-, –O-, and –NR. 2 - group.

[0180] In some implementations, ring B is a saturated 5-membered heterocycle with a structure And optionally includes another option selected from –NH-, -O- and –NR. 2 - group.

[0181] In some implementations, ring B is a saturated 6-membered heterocycle with a structure And optionally includes another option selected from –NH-, -O- and –NR. 2 -A group. In some embodiments, ring B is a group having a structure A saturated 6-membered heterocycle, wherein ring B further comprises another element selected from –NH-, -O-, and –NR-. 2 - group.

[0182] In some implementations, ring B is a saturated 7-membered heterocycle with a structure And optionally includes another option selected from –NH-, -O- and –NR. 2 -A group. In some embodiments, ring B is a group having a structure A saturated 7-membered heterocycle, wherein ring B further comprises another element selected from –NH-, -O-, and –NR-. 2 - group.

[0183] In some implementation schemes, ring B is selected from...

[0184]

[0185]

[0186]

[0187]

[0188] in:

[0189] R on nitrogen atom 2 Each for –(C 1-3 (Aliphatic group)R 3 ;and

[0190] R on carbon atom 2 Each is independently selected from halogens, N(R)2, OR, or –(C 1-3 (Aliphatic group)R 3 .

[0191] In some implementation schemes, ring B is selected from...

[0192]

[0193] Generally, as defined above, R is each independently a hydrogen or optionally substituted group, selected from C.1-6 An aliphatic group; a 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur; a phenyl group and a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur; or two R groups together with the nitrogen atom to which they are attached to form an optionally substituted 3- to 7-membered monocyclic heterocycle having 0 to 2 additional heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, R is hydrogen. In some embodiments, R is an optionally substituted group selected from C. 1-6 Aliphatic group, 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl and 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur; or two R groups together with the nitrogen atom to which they are attached to form an optionally substituted 3- to 7-membered monocyclic heterocycle having 0 to 2 additional heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0194] In some implementations, R is an optional substitute for C. 1-6 Aliphatic group. In some embodiments, R is a C group optionally substituted with oxo and OR°. 1-6 Aliphatic groups, where R° is C 1-6 Aliphatic group. In some such embodiments, R is –C(O)OtBu.

[0195] In some implementations, R is C 1-6 Aliphatic group. In some such embodiments, R is methyl or ethyl.

[0196] In some embodiments, R is selected from hydrogen and optionally substituted C. 1-6 Aliphatic group. In some such embodiments, R is selected from hydrogen, methyl, or ethyl.

[0197] Generally as defined above, R 2 Each can be independently a halogen, N(R)2, OR, or –(C 1-3 (Aliphatic group)R 3 In some implementations, R 2 It is a halogen. In some implementations, R 2 For N(R)². In some such implementations, R 2 For NH2. In some implementations, R 2 For OR. In some such implementations, R 2 It is OH.

[0198] In some implementation schemes, R 2 For –(C 1-3 (Aliphatic group)R 3 In some implementations, R2 For –CH2R 3 In some implementations, R 2 –CH(CH3)R 3 In some implementations, R 2 For –CH2CH2R 3 .

[0199] Generally as defined above, R 3 Each of the groups is an optional substituted group, selected from C. 1-6 Aliphatic group, 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl, 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, 8- to 10-membered bicyclic saturated, partially unsaturated or aryl carbon ring, 8- to 10-membered bicyclic saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, or 8- to 10-membered bicyclic heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0200] In some implementation schemes, R 3 C is an optional replacement 1-6 Aliphatic groups. In some such embodiments, R 3 The substituted group is optionally selected from cyclopentyl or cyclohexyl. In some embodiments, R 3 C 1-6 Aliphatic group. In some embodiments, R 3 It is methyl. In some embodiments, R 3 It is ethyl. In some embodiments, R 3 It is a cyclohexyl group.

[0201] In some implementation schemes, R 3 The phenyl group is optionally substituted.

[0202] In some implementation schemes, R 3 It is a 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3 It is a 3-membered saturated heterocycle with one optionally substituted heteroatom selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3 It is a 4-membered saturated heterocycle with one independently substituted heteroatom selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3 It is a 5-membered saturated or partially unsaturated heterocycle with 1-2 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3It is a 6-membered saturated or partially unsaturated heterocycle with 1-3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3 The substituted group is selected from pyrrolidinyl, piperidinyl, morpholinyl, and piperazineyl.

[0203] In some implementation schemes, R 3 It is a 5- to 6-membered heteroaryl ring with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3 It is a 5-membered heteroaryl ring with 1-3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3 It is a 5-membered heteroaryl ring with 1-2 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some such embodiments, R 3 The substituted group is selected from thienyl, pyrazolyl and pyrazolyl groups.

[0204] In some implementation schemes, R 3 It is an optionally substituted 6-membered heteroaryl ring having 1-3 nitrogen atoms. In some embodiments, R 3 It is an optionally substituted 6-membered heteroaryl ring having 1-2 nitrogen atoms. In some such embodiments, R 3 The substituted group is selected from pyridinyl or pyrimidinyl.

[0205] In some implementation schemes, R 3 The substituted 8- to 10-membered bicyclic saturated, partially unsaturated, or aryl carbocyclic rings are used. In some embodiments, R 3 The substituted 9-membered bicyclic saturated, partially unsaturated, or aryl carbocyclic ring is used. In some such embodiments, R 3 The substituted 2,3-dihydro-1H-indenyl group is used. In some embodiments, R 3 The substituted 10-membered bicyclic saturated, partially unsaturated, or aryl carbocyclic ring is used. In some such embodiments, R 3 The substituted group is selected from 1,2,3,4-tetrahydronaphthyl and naphthyl.

[0206] In some implementation schemes, R 3 It is an 8- to 10-membered bicyclic saturated or partially unsaturated heterocycle having 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3 It is a 9-membered bicyclic saturated or partially unsaturated heterocycle with 1-3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3It is a 10-membered bicyclic saturated or partially unsaturated heterocycle having 1-3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some such embodiments, R 3 The substituted group is selected from chromium, isochromium, 1,2,3,4-tetrahydroquinolinyl, 3,4-dihydro-2H-benzo[b][1,4]oxazinyl and 2H-benzo[b][1,4]oxazin-3(4H)-keto.

[0207] In some implementation schemes, R 3 It is an 8- to 10-membered bicyclic heteroaryl ring with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, R 3 It is a 9-membered bicyclic heteroaryl ring with 1-3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur. In some such embodiments, R 3 The substituted group is selected from indolyl, benzopyrazolyl, benzoimidazolyl and imidazo[1,2-a]pyridyl.

[0208] In some implementation schemes, R 3 Selected from

[0209]

[0210]

[0211] In some particularly preferred embodiments, R 3 Selected from

[0212] Therefore, in some implementation schemes, R 2 Selected from

[0213]

[0214] In some particularly preferred embodiments, R 3 Selected from

[0215]

[0216] Therefore, in some implementation schemes, R 2 Selected from

[0217]

[0218] In some implementations of Equation I, ring B is... Therefore, in some embodiments, this disclosure provides compounds of formula Ia:

[0219]

[0220] Or a pharmaceutically acceptable salt thereof, wherein rings A and R x R 1 R 2 Each of n is as defined above and described in this article.

[0221] In some implementations of Equation I, ring B is... Therefore, in some embodiments, this disclosure provides compounds of formula Ib:

[0222]

[0223] Or a pharmaceutically acceptable salt thereof, wherein rings A and R x R 1 R 2 Each of n is as defined above and described in this article.

[0224] In some implementations of Equation I, ring B is... Therefore, in some embodiments, this disclosure provides compounds of formula Ic:

[0225]

[0226] Or a pharmaceutically acceptable salt thereof, wherein rings A and R x R 1 R 2 Each of n is as defined above and described in this article.

[0227] In some implementations of Equation I, ring B is... Therefore, in some embodiments, this disclosure provides compounds of formula Id:

[0228]

[0229] Or a pharmaceutically acceptable salt thereof, wherein rings A and R x R 1 R 2 Each of n is as defined above and described in this article.

[0230] In some implementations of Equation I, ring B is... Therefore, in some embodiments, this disclosure provides compounds of formula Ie:

[0231]

[0232] Or a pharmaceutically acceptable salt thereof, wherein rings A and R x R 1 R 2 Each of n is as defined above and described in this article.

[0233] In some embodiments of Formula I, ring A is a 5-membered heteroaryl ring having 2-3 nitrogen atoms. Therefore, in some embodiments, this disclosure provides compounds of formula If, Ig, Ih, or Ii:

[0234]

[0235] Or a pharmaceutically acceptable salt thereof, wherein rings B and R 1 Each of them is as defined above and described herein.

[0236] In some implementation schemes, R 1 for Therefore, in some embodiments, this disclosure provides compounds of the formulas Iai, Ibi, Ici, Idi, Iei, Ifi, and Igi:

[0237]

[0238] Or a pharmaceutically acceptable salt thereof, wherein ring A, ring B, or ring R x R 2 Each of n is as defined above and described in this article.

[0239] In some implementations of equation Ib, ring A is... Therefore, in some embodiments, this disclosure provides compounds of formula Ib-ii:

[0240]

[0241] Or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 Each of them is as defined above and described herein.

[0242] In some implementations of formula Ib-ii, R 2 For –CH2R 3 Therefore, in some embodiments, this disclosure provides compounds of formulas Ij, Iji, and Ij-ii:

[0243]

[0244] Or a pharmaceutically acceptable salt thereof, wherein R 1 and R 3 Each of them is as defined above and described herein.

[0245] In one embodiment, this disclosure provides compounds of formula II:

[0246]

[0247] Or its pharmaceutically acceptable salt, wherein:

[0248] R 1 It is a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms selected from oxygen, nitrogen, and sulfur;

[0249] G represents CH and CR. x Or N;

[0250] R x It is a C1-C3 alkyl, halogen, or cyano group;

[0251] X is CH2, NH, N (C1-C3 alkyl) or O;

[0252] Y is C(R) p )2 or NH;

[0253] Z represents a valence bond, CH2, or -CH2CH2-;

[0254] R 2a -(C1-C3 alkyl)R 3 ;

[0255] R 2b It is hydrogen, halogen, C1-C3 alkyl or -(C1-C3 alkyl)R 3 ;

[0256] R p Independently hydrogen, halogen, or NH2;

[0257] R 3 It is a benzene ring or a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms selected from oxygen, nitrogen, and sulfur, wherein the aryl ring or the 5- to 6-membered heteroaryl ring is optionally surrounded by 1 to 2 R atoms. q replace;

[0258] R q It can be halogen, cyano, or -CF3.

[0259] In another embodiment, this disclosure provides a compound of formula II as described above, wherein R 2b It is hydrogen.

[0260] In another embodiment, this disclosure provides a compound of formula II as described above, wherein R 1 Selected from

[0261] In another embodiment, this disclosure provides compounds of formula II as described above, wherein X is CH2, Y is CH2 and Z is CH2.

[0262] In another embodiment, this disclosure provides a compound of formula II as described above, wherein R 2a -CH2-R 3 .

[0263] In another embodiment, this disclosure provides a compound of formula II as described above, wherein R 3 Selected from

[0264]

[0265]

[0266] In another embodiment, this disclosure provides a compound of formula II as described above, wherein R 3 Selected from

[0267] In another embodiment, this disclosure provides a compound of formula II as described above, which is

[0268]

[0269] Or its pharmaceutically acceptable salt.

[0270] In another embodiment, this disclosure provides a compound of formula II as described above, which is

[0271]

[0272] Or its pharmaceutically acceptable salt.

[0273] In another embodiment, this disclosure provides a compound of formula II as described above, which is

[0274]

[0275] Or its pharmaceutically acceptable salt.

[0276] In another embodiment, this disclosure provides a compound of formula II as described above, which is

[0277]

[0278] Or its pharmaceutically acceptable salt.

[0279] In some embodiments, this disclosure provides compounds selected from the following:

[0280]

[0281]

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300] Or its pharmaceutically acceptable salt.

[0301] In some aspects, this disclosure provides compounds with the following embodiments:

[0302] Implementation Scheme 1: Compound of Formula I:

[0303]

[0304] Or its pharmaceutically acceptable salt, wherein:

[0305] Ring A is a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur;

[0306] R 1The substituted group is selected from a 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, or a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0307] R x Each group is independently selected from halogen, cyano, OR, SR, N(R)2, or optionally substituted groups, and is selected from C. 1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbon rings, 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur;

[0308] Ring B is a saturated 5- to 7-membered heterocycle with a structure And optionally includes another option selected from –NH-, -O- and –NR. 2 - group;

[0309] R is each independently hydrogen or an optionally substituted group, selected from C 1-6 Aliphatic group, 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur, phenyl, and 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur; or:

[0310] The two R groups together with the nitrogen atom to which they are attached form a 3- to 7-membered monocyclic heterocycle with 0-2 additional heteroatoms independently selected from oxygen, nitrogen and sulfur;

[0311] R 2 Each can be independently a halogen, N(R)2, OR, or -(C) 1-3 (Aliphatic group)R 3 ;

[0312] R 3 Each of the groups is an optional substituted group, selected from C. 1-6 Aliphatic group, 3- to 7-membered saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl, 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, 8- to 10-membered bicyclic saturated, partially unsaturated or aryl carbon ring, 8- to 10-membered bicyclic saturated or partially unsaturated heterocycle having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, or 8- to 10-membered bicyclic heteroaryl ring having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur;

[0313] m is 0, 1, or 2; and

[0314] n is 0, 1, or 2.

[0315] Implementation Scheme 2: The compound of Implementation Scheme 1, wherein ring A is a 6-membered heteroaryl ring having 1-2 nitrogen atoms.

[0316] Implementation Scheme 3: The compound of Implementation Scheme 1, wherein ring A is a 5-membered heteroaryl ring having 1-3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0317] Implementation Scheme 4: The compound of Implementation Scheme 3, wherein ring A is a 5-membered heteroaryl ring having two heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0318] Implementation Scheme 5: The compound of Implementation Scheme 3, wherein ring A is a 5-membered heteroaryl ring having three heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0319] Implementation Scheme 6: The compound of Implementation Scheme 1, wherein ring A is selected from...

[0320]

[0321] Implementation Scheme 7: The compound of Implementation Scheme 6, wherein ring A is selected from...

[0322] Implementation Scheme 8: The compound of Implementation Scheme 1, wherein ring A is selected from...

[0323]

[0324]

[0325] Implementation Scheme 9: The compound of Implementation Scheme 1, wherein ring A is selected from...

[0326]

[0327] Where R x The substituted group is selected from C. 1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbon rings, 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl groups, and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0328] Implementation Scheme 10: The compound of Implementation Scheme 1, wherein ring A is selected from...

[0329]

[0330] Implementation Scheme 11: The compound of Implementation Scheme 1, wherein ring A is selected from...

[0331]

[0332] in:

[0333] R on nitrogen atom x The groups selected from the optionally substituted groups are selected from C. 1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbocyclic rings, 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur, phenyl groups, and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen, and sulfur; and

[0334] R on carbon atom x Selected from halogen, cyano, OR, SR, N(R)2, or optionally substituted groups, wherein the group is selected from C 1-4 Aliphatic groups, 3- to 7-membered saturated or partially unsaturated carbon rings, 3- to 7-membered saturated or partially unsaturated heterocycles having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, phenyl groups, and 5- to 6-membered heteroaryl rings having 1 to 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0335] Implementation Scheme 12: A compound of any one of Implementation Schemes 1-11, wherein R x It is a halogen.

[0336] Implementation Scheme 13: A compound of any one of Implementation Schemes 1-11, wherein R x It is a cyano group.

[0337] Implementation Scheme 14: A compound of any one of Implementation Schemes 1-11, wherein R x OR.

[0338] Implementation Scheme 15: A compound of any one of Implementation Schemes 1-11, wherein R x It is SR.

[0339] Implementation Scheme 16: A compound of any one of Implementation Schemes 1-11, wherein R x It is N(R)2.

[0340] Implementation Scheme 17: Compounds 14-16 of any one of the implementation schemes, wherein R is selected from hydrogen and optionally substituted C. 1-6 Aliphatic groups.

[0341] Implementation Scheme 18: The compound of Implementation Scheme 17, wherein R is selected from hydrogen and optionally substituted C. 1-4 Aliphatic groups.

[0342] Implementation Scheme 19: A compound of any one of Implementation Schemes 14, 17 and 18, wherein R x The values ​​are OH, OCH3, and OCH2CH3.

[0343] Implementation Scheme 20: A compound of any one of Implementation Schemes 15, 17 and 18, wherein R x For SH, SCH3, and SCH2CH3.

[0344] Implementation Scheme 21: A compound of any one of Implementation Schemes 16-18, wherein R x Selected from NH2, NHCH3, NHCH2CH3, N(CH3)2 and N(CH2CH3)2.

[0345] Implementation Scheme 22: A compound of any one of Implementation Schemes 1-11, wherein R x C is an optional replacement 1-4 Aliphatic groups.

[0346] Implementation Scheme 23: The compound of Implementation Scheme 22, wherein R x C is an optional replacement 3-4 Aliphatic groups.

[0347] Implementation Scheme 24: The compound of Implementation Scheme 23, wherein R x Selected from tert-butyl,

[0348] Implementation Scheme 25: The compound of Implementation Scheme 22, wherein R x C 1-4 Aliphatic groups, optionally substituted with groups selected from: halogens, –(CH2). 0–4 R°、–(CH2) 0–4 OR°、-(CH2) 0–4 N(R°)2、–(CH2) 0–4 C(O)OR° and –(CH2) 0– 4C(O)NR°2.

[0349] Implementation Scheme 26: The compound of Implementation Scheme 25, wherein R° is selected from hydrogen, C 1–6 Aliphatic groups, –CH2Ph, –O(CH2) 0–1 Ph, -CH2- (5- to 6-membered heteroaryl ring), 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or 8- to 10-membered bicyclic aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or: two independently occurring R° together with their intervening atoms forming a 3- to 12-membered saturated, partially unsaturated or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0350] Implementation Scheme 27: The compound of Implementation Scheme 22, wherein R x C 1-4Aliphatic groups, which are optionally substituted with groups selected from the following: halogen, –R°, –OR°, -N(R°)2, –C(O)OR° and –C(O)NR°2.

[0351] Implementation Scheme 28: The compound of Implementation Scheme 22, wherein R x C that is optionally substituted with halogen 1-4 Aliphatic groups.

[0352] Implementation Scheme 29: The compound of Implementation Scheme 28, wherein R x Selected from –CH3, –CF3, -CHF2 and CH2F.

[0353] Implementation Scheme 30: The compound of Implementation Scheme 22, wherein R x Selected from –CH2R°, –CH2OR°, –CH2N(R°)2, –CH2C(O)OR° and –CH2C(O)N(R°)2.

[0354] Implementation Scheme 31: The compound of Implementation Scheme 30, wherein R x Selected from –CH2OH, -CH2OCH3, -CH2C(O)NH2, -CH2C(O)NHCH3 and -CH2C(O)N(CH3)2.

[0355] Implementation Scheme 32: A compound of any one of Implementation Schemes 1-31, wherein R 1 It is a 5- to 6-membered heteroaryl ring with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen and sulfur.

[0356] Implementation Scheme 33: The compound of Implementation Scheme 32, wherein R 1 It is a 5-membered heteroaryl ring with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen and sulfur.

[0357] Implementation Scheme 34: The compound of Implementation Scheme 33, wherein R 1 It is a 5-membered heteroaryl ring with 1-2 independently substituted heteroatoms selected from oxygen, nitrogen and sulfur.

[0358] Implementation Scheme 35: The compound of Implementation Scheme 32, wherein R 1 It is a 6-membered heteroaryl ring with 1-3 nitrogen atoms that are optionally substituted.

[0359] Implementation Scheme 36: The compound of Implementation Scheme 35, wherein R 1 It is a 6-membered heteroaryl ring with 1-2 nitrogen atoms that are optionally substituted.

[0360] Implementation Scheme 37: The compound of Implementation Scheme 32, wherein R 1 Selected from

[0361]

[0362] Implementation Scheme 38: The compound of Implementation Scheme 37, wherein R 1 Selected from

[0363] Implementation Scheme 39: A compound of any one of Implementation Schemes 1-38, wherein ring B is a saturated 5-membered heterocycle, having the structure And optionally includes another option selected from –NH-, -O- and –NR. 2 - group.

[0364] Implementation Scheme 40: A compound of any one of Implementation Schemes 1-38, wherein ring B is a saturated 6-membered heterocycle, having the structure And optionally includes another option selected from –NH-, -O- and –NR. 2 - group.

[0365] Implementation Scheme 41: The compound of Implementation Scheme 40, wherein ring B is a compound having the structure A saturated 6-membered heterocycle, wherein ring B further comprises another element selected from –NH-, -O-, and –NR-. 2 - group.

[0366] Implementation Scheme 42: A compound of any one of Implementation Schemes 1-38, wherein ring B is a saturated 7-membered heterocycle, having the structure And optionally includes another option selected from –NH-, -O- and –NR. 2 - group.

[0367] Implementation Scheme 43: The compound of Implementation Scheme 42, wherein ring B is a compound having the structure A saturated 7-membered heterocycle, wherein ring B further comprises another element selected from –NH-, -O-, and –NR-. 2 - group.

[0368] Implementation Scheme 44: A compound of any one of Implementation Schemes 1-38, wherein ring B is selected from...

[0369]

[0370]

[0371]

[0372]

[0373] Implementation Scheme 45: The compound of Implementation Scheme 44, wherein ring B is selected from...

[0374]

[0375] Implementation Scheme 46: A compound of any one of Implementation Schemes 1-45, wherein R 2 For –(C 1-2 (Aliphatic group)R 3 .

[0376] Implementation Scheme 47: The compound of Implementation Scheme 46, wherein R 2 For –CH2R 3 .

[0377] Implementation Scheme 48: The compound of Implementation Scheme 46, wherein R 2 –CH(CH3)R 3 .

[0378] Implementation Scheme 49: The compound of Implementation Scheme 46, wherein R 2 For –CH2CH2R 3 .

[0379] Implementation Scheme 50: A compound of any one of Implementation Schemes 1-49, wherein R 3 The phenyl group is optionally substituted.

[0380] Implementation Scheme 51: A compound of any one of Implementation Schemes 1-49, wherein R 3 It is a 5- to 6-membered heteroaryl ring with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen and sulfur.

[0381] Implementation Scheme 52: The compound of Implementation Scheme 51, wherein R 3 It is a 5-membered heteroaryl ring with 1-2 independently substituted heteroatoms selected from oxygen, nitrogen and sulfur.

[0382] Implementation Scheme 53: The compound of Implementation Scheme 51 or 52, wherein R 3 The substituted group is selected from thienyl, pyrazolyl, and imidazolyl.

[0383] Implementation Scheme 54: The compound of Implementation Scheme 51, wherein R 3 It is a 6-membered heteroaryl ring with 1-3 nitrogen atoms that are optionally substituted.

[0384] Implementation Scheme 55: The compound of Implementation Scheme 54, wherein R 3 It is a 6-membered heteroaryl ring with 1-2 nitrogen atoms that are optionally substituted.

[0385] Implementation Scheme 56: The compound of Implementation Scheme 55, wherein R 3 The substituted group is selected from pyridinyl or pyrimidinyl.

[0386] Implementation Scheme 57: A compound of any one of Implementation Schemes 1-49, wherein R 3 It is an substituted 8- to 10-membered bicyclic saturated, partially unsaturated, or aryl carbocyclic ring.

[0387] Implementation Scheme 58: The compound of Implementation Scheme 57, wherein R 3 It is a 9-membered bicyclic saturated, partially unsaturated, or aryl carbide ring with optional substitution.

[0388] Implementation Scheme 59: The compound of Implementation Scheme 58, wherein R 3 It is an optionally substituted 2,3-dihydro-1H-indenyl group.

[0389] Implementation Scheme 60: The compound of Implementation Scheme 57, wherein R 3 It is an optionally substituted 10-membered bicyclic saturated, partially unsaturated, or aryl carbocyclic ring.

[0390] Implementation Scheme 61: The compound of Implementation Scheme 60, wherein R 3 The substituted group is selected from 1,2,3,4-tetrahydronaphthyl or naphthyl.

[0391] Implementation Scheme 62: A compound of any one of Implementation Schemes 1-49, wherein R 3 It is an 8- to 10-membered bicyclic saturated or partially unsaturated heterocycle with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur.

[0392] Implementation Scheme 63: The compound of Implementation Scheme 62, wherein R 3 It is a 9-membered bicyclic saturated or partially unsaturated heterocycle with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen and sulfur.

[0393] Implementation Scheme 64: The compound of Implementation Scheme 62, wherein R 3 It is a 10-membered bicyclic saturated or partially unsaturated heterocycle with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen and sulfur.

[0394] Implementation Scheme 65: The compound of Implementation Scheme 64, wherein R 3 The substituted group is selected from chromium, isochromium, 1,2,3,4-tetrahydroquinolinyl, 3,4-dihydro-2H-benzo[b][1,4]oxazinyl and 2H-benzo[b][1,4]oxazin-3(4H)-keto.

[0395] Implementation Scheme 66: A compound of any one of Implementation Schemes 1-49, wherein R 3 It is an 8- to 10-membered bicyclic heteroaryl ring with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen, and sulfur.

[0396] Implementation Scheme 67: The compound of Implementation Scheme 66, wherein R 3 It is a 9-membered bicyclic heteroaryl ring with 1 to 3 independently substituted heteroatoms selected from oxygen, nitrogen and sulfur.

[0397] Implementation Scheme 68: The compound of Implementation Scheme 66 or 67, wherein R 3 The substituted group is selected from indolyl, benzopyrazolyl, benzoimidazolyl and imidazo[1,2-a]pyridyl.

[0398] Implementation Scheme 69: A compound of any one of Implementation Schemes 1-49, wherein R 3 Selected from

[0399]

[0400]

[0401]

[0402] Implementation Scheme 70: The compound of Implementation Scheme 69, wherein R 3 Selected from

[0403]

[0404] Implementation Scheme 70a: The compound of Implementation Scheme 69, wherein R 3 Selected from

[0405]

[0406] Implementation Scheme 71: The compound of Implementation Scheme 1, wherein the compound is:

[0407]

[0408] Or its pharmaceutically acceptable salt.

[0409] Implementation Scheme 72: The compound of Implementation Scheme 1, wherein the compound is:

[0410]

[0411] Or its pharmaceutically acceptable salt.

[0412] Implementation Scheme 73: The compound of Implementation Scheme 1, wherein the compound is:

[0413]

[0414] Or its pharmaceutically acceptable salt.

[0415] Implementation Scheme 74: The compound of Implementation Scheme 1, wherein the compound is:

[0416]

[0417] Or its pharmaceutically acceptable salt.

[0418] Implementation Scheme 75: The compound of Implementation Scheme 1, wherein the compound is:

[0419]

[0420] Or its pharmaceutically acceptable salt.

[0421] Implementation Scheme 76: The compound of Implementation Scheme 1, wherein the compound is selected from:

[0422]

[0423] Or its pharmaceutically acceptable salt.

[0424] Implementation Scheme 77: The compound of Implementation Scheme 1, wherein the compound is selected from:

[0425]

[0426]

[0427] Or its pharmaceutically acceptable salt.

[0428] Implementation Scheme 77a: A compound of any one of Implementation Schemes 1, 3, 5, 6, 7, 8, 32, 35, 36, 37, 38, 40, 44, 45, 46, 47, 51, 54, 55, 56, 69, 70a, and 72, wherein ring A is

[0429] Implementation Scheme 77b: The compound of Implementation Scheme 77a, wherein the compound is

[0430]

[0431] Or its pharmaceutically acceptable salt.

[0432] Implementation Scheme 77c: The compound of Implementation Scheme 77b, wherein the compound is selected from:

[0433]

[0434] Or its pharmaceutically acceptable salt.

[0435] Implementation Scheme 77d: The compound of Implementation Scheme 77c, wherein R 1 for

[0436] Implementation Scheme 77e: The compound of Implementation Scheme 77d, wherein R 3 The pyridinyl group is optionally substituted.

[0437] Implementation Scheme 77f: The compound of Implementation Scheme 77e, wherein R 3 Selected from

[0438] Implementation Scheme 77g: A compound of any one of Implementation Schemes 1, 77a, 77b, 77c, 77d, 77e, or 77f, wherein the compound is

[0439]

[0440] Or its pharmaceutically acceptable salt.

[0441] Implementation scheme 77h: A compound of any one of implementation schemes 1, 77a, 77b, 77c, 77d, 77e, or 77f, wherein the compound is

[0442]

[0443] Or its pharmaceutically acceptable salt.

[0444] Implementation Scheme 78: A pharmaceutical composition comprising a compound of any one of Implementation Schemes 1-77h and a pharmaceutically acceptable carrier.

[0445] Implementation Scheme 79: A method comprising the following steps: administering a compound of any one of Implementation Schemes 1-77h to a subject, wherein the subject (i) has a condition characterized by axonal degeneration or (ii) is at risk of developing a condition characterized by axonal degeneration.

[0446] Implementation Plan 80: A method for treating or preventing axonal degeneration, including administering a compound of any one of Implementation Plans 1-77h to a subject in need of such treatment.

[0447] Implementation Scheme 81: A method for inhibiting SARM1, comprising exposing a biological sample to the compound of any one of Implementation Schemes 1-77h.

[0448] Composition

[0449] In some embodiments, the compound of formula I may be provided in the composition, for example, in combination with one or more other ingredients (e.g., a mixture).

[0450] In some embodiments, this disclosure provides compositions comprising and / or delivering a compound of formula I or its active metabolite, for example, when in contact with or otherwise applied to a system or environment, such as a system or environment that may include SARM1 NAD enzyme activity; in some embodiments, application of such compositions to a system or environment achieves inhibition of SARM1 activity as described herein.

[0451] In some embodiments, the compositions provided herein may be pharmaceutical compositions because they contain an active agent and one or more pharmaceutically acceptable excipients; in some such embodiments, the pharmaceutical compositions provided contain and / or deliver a compound of formula I or its active metabolite to the relevant system or environment as described herein (e.g., to a subject in need of it).

[0452] In some embodiments, one or more compounds of formula I are provided and / or utilized in the form of a pharmaceutically acceptable salt.

[0453] This disclosure provides compositions comprising a compound of formula I or a pharmaceutically acceptable salt or derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or mediator. The amount of the compound in the provided compositions is such that axonal degeneration is effectively and measurably inhibited in biological samples or patients. In some embodiments, the provided compounds or compositions are formulated for administration to a patient requiring such a composition. The compounds and compositions of the methods of this disclosure can be administered in any amount and via any route of administration for the effective treatment or reduction of the severity of any disease or condition described herein. The provided compounds are preferably formulated in dosing unit form for ease of administration and uniform dosing. As used herein, “dosing unit form” refers to a physically discrete unit of the agent suitable for the patient to be treated. However, it should be understood that the total daily dose of the provided compounds and compositions will be determined by the attending physician within reasonable medical judgment. The specific effective dose level for any particular patient or organism will vary from subject to subject to a variety of factors, including the condition being treated and its severity; the activity of the specific compound used; the specific composition used and its route of administration; the patient's species, age, weight, sex, and diet; the subject's general condition; the time of administration; the excretion rate of the specific compound used; the duration of treatment; and the drugs used in combination with or concurrently with the specific compound used.

[0454] The provided composition can be administered orally, parenterally, by inhalation or nasal spray, topically (e.g., by powder, ointment, or drops), rectally, orally, intravaginally, intraperitoneally, intracisionally, or via an implanted reservoir, depending on the severity of the condition being treated. Preferably, the composition is administered orally, intraperitoneally, or intravenously. In some embodiments, the provided compound is administered orally or parenterally at a dose level of about 0.01 mg / kg to about 50 mg / kg of the subject's body weight once or more daily to achieve the desired therapeutic effect.

[0455] As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrasheath, intrahepatic, intralesional, and intracranial injection or infusion techniques. The sterile injectable form of the provided compositions may be an aqueous or oily suspension. These suspensions may be formulated using techniques known in the art, employing suitable dispersants or wetting agents and suspending agents. Sterile injectable formulations may also be sterile injectable solutions or suspensions in non-toxic, parenteral-acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable media and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. Furthermore, sterile, non-volatile oils are commonly used as solvents or suspension media.

[0456] For this purpose, any mild, non-volatile oil can be used, including synthetic mono- or diglycerides of glycerol. Fatty acids such as oleic acid and its glycerol derivatives can be used to prepare injectable formulations, as can natural, pharmaceutically acceptable oils such as olive oil or castor oil, especially their polyoxyethylated forms. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as carboxymethyl cellulose or similar dispersants commonly used in the formulation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tween, Span, and other emulsifiers or bioavailability enhancers commonly used in the preparation of pharmaceutically acceptable solid, liquid, or other dosage forms, may also be used for formulation purposes.

[0457] Injectable formulations can be sterilized, for example, by filtering through a bacterial trap or by incorporating a sterilizing agent in the form of a sterile solid composition, which can be dissolved or dispersed in sterile water or other sterile injectable media prior to use.

[0458] To prolong the effect of the provided compound, it is often necessary to slow its absorption from subcutaneous or intramuscular injection. This can be achieved by using liquid suspensions of poorly water-soluble crystalline or amorphous materials. The absorption rate of the compound then depends on its dissolution rate, which in turn depends on the crystal size and crystal form. Alternatively, delayed absorption of the parenteral administration of the compound can be achieved by dissolving or suspending the compound in an oil medium. Injectable reservoir forms are prepared by forming microcapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolic acid. The rate of compound release can be controlled depending on the ratio of compound to polymer and the properties of the specific polymer used. Other examples of biodegradable polymers include poly(orthoester) and poly(anhydride). Injectable reservoir formulations are also prepared by encapsulating the compound in liposomes or microemulsions that are compatible with body tissues.

[0459] The pharmaceutically acceptable compositions described herein can be administered orally in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions, or solutions. In such solid dosage forms, the active compound can be mixed with at least one inert diluent such as sucrose, lactose, or starch. As is normal practice, such dosage forms may also contain substances other than inert diluents, such as lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. When an aqueous suspension is required for oral administration, the active ingredient is combined with emulsifiers and suspending agents. If desired, certain sweeteners, flavoring agents, or coloring agents may also be added.

[0460] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier (such as sodium citrate or dicalcium phosphate) and / or the following components: a) fillers or enrichments, such as starch, lactose, sucrose, glucose, mannitol, and silica; b) binders, such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; c) humectants, such as glycerin; d) disintegrants, such as agar, calcium carbonate, potato or cassava starch, alginate, certain silicates, and sodium carbonate; e) solution blockers, such as paraffin; f) absorption enhancers, such as quaternary ammonium compounds; g) wetting agents, such as cetyl alcohol and glyceryl monostearate; h) absorbents, such as kaolin and bentonite; and / or i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain a buffer. The active compound may also be in a microencapsulated form having one or more excipients as described above.

[0461] Similar types of solid compositions can also be used as fillers in soft and hard-filled gelatin capsules, which use excipients such as lactose or milk sugar and high molecular weight polyethylene glycol. Solid dosage forms of tablets, sugar-coated pills, capsules, pellets, and granules can be prepared using coatings and shells, such as enteric coatings (i.e., buffers) and other coatings well known in the pharmaceutical formulation field. They may optionally contain light-blocking agents and may also be compositions that release the active ingredient only or preferentially in a portion of the intestine, optionally in a delayed manner. Examples of encapsulation compositions that can be used include polymeric substances and waxes.

[0462] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, liquid dosage forms may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, dimethylformamide, oils (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitol and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and aromatizers.

[0463] Alternatively, the pharmaceutically acceptable compositions described herein can be administered in the form of suppositories for rectal or vaginal application. These can be prepared by mixing the compounds of this disclosure with a suitable non-irritating excipient or carrier that is solid at room temperature but liquid at body (e.g., rectal or vaginal) temperature, and thus melts in the rectal or vaginal cavity to release the active compound. Such materials include cocoa butter, suppository waxes (e.g., beeswax), and polyethylene glycol.

[0464] The pharmaceutically acceptable compositions described herein can also be applied topically, particularly when the treatment target includes areas or organs easily accessible for topical application, including eye, skin, or lower bowel diseases. Topical application to the lower bowel can be achieved in the form of rectal suppositories (see above) or in suitable enema formulations.

[0465] Dosage forms of compounds intended for topical or transdermal application include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalers, or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any desired preservatives or buffers that may be required. Ophthalmic preparations, ear drops, and eye drops are also considered within the scope of this disclosure. Additionally, this disclosure contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of the compound to the body. Such dosage forms can be prepared by dissolving or dispensing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by providing a rate-controlled membrane or by dispersing the compound in a polymer matrix or gel.

[0466] For topical application, the pharmaceutically acceptable compositions provided can be formulated into suitable ointments containing an active ingredient suspended or dissolved in one or more carriers. Carriers for topical application of the compounds disclosed herein include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsified waxes, and water. Alternatively, the pharmaceutically acceptable compositions provided can be formulated into suitable lotions or creams containing an active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl wax, cetearyl alcohol, 2-octyldodecanool, benzyl alcohol, and water.

[0467] For ophthalmic use, the pharmaceutically acceptable composition provided may be formulated as a micronized suspension in isotonic, pH-adjusted sterile saline, or preferably as a solution in isotonic, pH-adjusted sterile saline, with or without preservatives, such as benzalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutically acceptable composition may be formulated in an ointment such as petrolatum.

[0468] The pharmaceutically acceptable compositions disclosed herein can also be administered via nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the field of pharmaceutical formulation and can be prepared as solutions in saline, using benzyl alcohol or other suitable preservatives, bioavailability enhancers, fluorocarbons, and / or other conventional solubilizers or dispersants.

[0469] Most preferably, the pharmaceutically acceptable composition of this disclosure is formulated for oral administration.

[0470] Identification and / or characterization of compounds and / or compositions

[0471] This disclosure provides various techniques for identifying and / or characterizing the compounds and / or compositions described herein. For example, this disclosure provides various assays for evaluating SARM1 inhibitory activity, and in particular for evaluating SARM1 inhibitory activity.

[0472] In some embodiments, the performance of one or more compounds or compositions of interest in the assays described herein is compared to the performance of a suitable reference. For example, in some embodiments, the reference may not be available for the relevant compound or composition. Or, additionally, in some embodiments, the reference may be available for alternative compounds or compositions, for example, those that have known performance in the relevant assays (e.g., as a positive or negative control, as understood in the art). In some embodiments, the reference may be an alternative but equivalent set of conditions (e.g., temperature, pH, salt concentration, etc.). In some embodiments, the reference may be the performance of the compound or composition with respect to the SARM1 variant.

[0473] Furthermore, alternatively or additionally, in some embodiments, the performance of one or more compounds or compositions of interest in the determinations described herein can be evaluated in the presence of a suitable reference compound or composition, for example, to determine the ability of a compound or composition to compete with a reference.

[0474] In some embodiments, multiple compounds or compositions of interest can be analyzed in a specific assay and / or compared with the same reference. In some embodiments, such multiple compounds or compositions can be or include a group of compounds or compositions considered a “library” because multiple members share one or more characteristics (e.g., structural units, source identity, synthetic similarity, etc.).

[0475] The following examples illustrate certain exemplary assays that can be used to implement this disclosure. Those skilled in the art will recognize that useful or relevant systems for identifying and / or characterizing compounds and / or compositions according to this disclosure are not limited to those included in the examples or otherwise discussed below.

[0476] In some embodiments, the compound and / or composition may be identified and / or characterized based on one or more activities or characteristics, such as promoting axonal integrity, cytoskeleton stability, and / or neuronal survival. In some embodiments, the provided SARM1 inhibitor inhibits the catabolism of NAD+ by SARM1. In some embodiments, the provided SARM1 inhibitor slows the rate of NAD+ catabolism.

[0477] In some embodiments, the provided SARM1 inhibitor reduces or inhibits the binding of NAD+ to SARM1. In some embodiments, the provided SARM1 inhibitor binds to SARM1 within a pouch containing one or more catalytic residues (e.g., a catalytic crack of SARM1). Examples of such catalytic residues include glutamate at position 642 (E642).

[0478] In some embodiments, the provided SARM1 inhibitor disrupts and / or prevents the polymerization of the TIR1 domain of SARM1. In some embodiments, the provided SARM1 inhibitor disrupts the polymerization of the SAM domain. In some embodiments, the disruption of the provided SARM1 inhibitor results in a NAD+-depleted axonal signaling cascade.

[0479] In some embodiments, this disclosure provides assays that can be used to identify and / or characterize one or more activities and / or features of compounds and / or compositions of interest. For example, in some embodiments, this disclosure provides in vitro, cellular, and / or in vivo systems for evaluating one or more such activities and / or features.

[0480] SARM1 activity assay

[0481] In some embodiments, the method for identifying SARM1 inhibitors includes: a) providing a mixture comprising i) a mutant or fragment of SARM1, ii) NAD+ and iii) a candidate inhibitor, wherein the mutant or fragment has constitutive activity; b) incubating the mixture; c) quantifying NAD+ in the mixture after the incubation; and d) identifying the candidate inhibitor compound as an inhibitor if the amount of NAD+ is greater than the amount of NAD+ in a control mixture that does not contain the candidate inhibitor.

[0482] In some embodiments, a method for identifying SARM1 inhibitors is provided, comprising: a) providing a mixture containing i) full-length SARM1, ii) NAD+ and iii) a candidate inhibitor, wherein the full-length SARM1 has constitutive activity; b) incubating the mixture; c) quantifying NAD+ and ADPR (or cADPR) in the mixture after incubation; d) determining the molar ratio of NAD+:ADPR (or cADPR); and e) identifying the candidate inhibitor compound as an inhibitor if the molar ratio is greater than that of a control mixture containing the candidate inhibitor.

[0483] In some embodiments, a method for identifying SARM1 inhibitors is provided, the method comprising: a) providing a mixture comprising a solid support incorporating i) full-length SARM1 and at least one tag, ii) NAD+, and iii) a candidate inhibitor; b) incubating the mixture; c) quantifying the NAD+ after the incubation; and d) identifying the candidate inhibitor compound as a SARM1 inhibitor if the concentration of NAD+ is greater than that of a control.

[0484] SARM1 binding assay

[0485] In some embodiments, the efficacy of the provided SARM1 inhibitor can be determined according to, for example, the assay described in WO 2018 / 057989, published March 29, 2018, which is incorporated herein by reference in its entirety. In some embodiments, the provided SARM1 inhibitor can be administered to a solution containing SARM1 or a fragment thereof. In some embodiments, the provided SARM1 inhibitor can be used in an in vitro system. In some embodiments, the provided SARM1 inhibitor can be used in vivo. In some embodiments, the provided SARM1 inhibitor can be administered to a patient. In some embodiments, the SARM1 inhibitor can be mixed with SARM1 or a fragment thereof that has been labeled with an epitope tag. In some embodiments, the affinity for the SARM1 inhibitor can be obtained by comparing the amount of bound SARM1 inhibitor with the amount of unbound SARM1 inhibitor.

[0486] In some embodiments, mutants or fragments of SARM1 are constitutively active SAM-TIR fragments. Constitutively active SARM1 fragments include, for example, but not limited to, SARM1 lacking an autorepressor domain; at least one point mutation in SARM1 inactivating said autorepressor domain; SARM1 fragments containing a TIR domain; or SARM1 fragments consisting of SAM and TIR domains. In some embodiments, the SARML peptide may include one or more additional amino acid sequences that can act as tags, such as His tags, streptavidin tags, or combinations thereof. In some embodiments, the SARML peptide may include tags at the N-terminus, C-terminus, or combinations thereof. In some embodiments, SARML or fragments thereof labeled with epitope tags can be used to measure the binding efficacy of the provided SARML inhibitor. Purification of the SARM1-TIR domain

[0487] In some embodiments, the SARM1-TIR domain can be modified with various proteins or epitope tags that can be used, for example, for purification. In some embodiments, this disclosure also provides an NRK1-HEK293T cell line comprising HEK293T cells transformed with nicotinamide nucleoside kinase 1 (NRK1). In some embodiments, HEK293T cells are transformed or transfected with a DNA sequence encoding nicotinamide nucleoside kinase 1 (Nrk1). In some embodiments, the DNA encoding NRK1 can be genomic DNA or cDNA. In some embodiments, HEK293T cells are stably or transiently transfected with DNA encoding NRK1 from an exogenous source derived from the host cell. In some embodiments, HEK293T cells are stably or transiently transfected with DNA encoding NRK1 such that the cells express NRK1 at an elevated level compared to control cells. In some embodiments, the DNA encoding NRK1 is under the control of one or more exogenous regulatory DNA sequences, such as promoters, enhancers, or combinations thereof. In some embodiments, the combination of the DNA sequence encoding NRK1 and the regulatory sequence is a non-naturally occurring combination. In some embodiments, the DNA (genomic or cDNA) encoding NRK1 comprises an expression vector, such as an FCIV expression vector. In some embodiments, the DNA encoding NRK1 is derived from cDNA or genomic DNA from vertebrate or invertebrate species (e.g., but not limited to humans, mice, zebrafish, or fruit flies). In some configurations, the NRK1 DNA is human NRK1 DNA.

[0488] Applications and uses

[0489] This disclosure provides for a variety of uses and applications of the compounds and / or compositions described herein, for example, based on their activities and / or characteristics as described herein. In some embodiments, such uses may include therapeutic and / or diagnostic uses. Alternatively, in some embodiments, such uses may include research, manufacturing, and / or other technical uses.

[0490] In one aspect, the present invention provides a method comprising administering one or more compounds of Formula I to a subject, for example, to treat, prevent, or reduce the risk of developing one or more conditions characterized by axonal degeneration. In some such embodiments, the compound of Formula I is a SARM1 inhibitor.

[0491] Another embodiment of this disclosure relates to a method for inhibiting SARM1 activity in a patient, comprising the step of administering the provided compound or a composition containing the compound to the patient.

[0492] Inhibiting enzymes in biological samples can be used for a variety of purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, biological assays, gene expression studies, and biological target identification.

[0493] In some embodiments, this disclosure relates to a method for treating axonal degeneration in a biological sample, comprising the step of contacting the biological sample with a compound or composition of Formula I. In some embodiments, one or more compounds and / or compositions as described herein can be used, for example, as a method for inhibiting neuronal degradation originating from a subject. In some embodiments, one or more compounds and / or compositions as described herein can be used to inhibit degeneration of neurons or portions thereof cultured in vitro. In some embodiments, one or more compounds and / or compositions as described herein can be used as stabilizers to promote neuronal survival in vitro.

[0494] In some embodiments, the provided compounds and / or compositions inhibit the NAD enzyme activity of SARM1. Optionally or additionally, in some embodiments, the compounds are provided to alleviate one or more properties of neurodegeneration. In some embodiments, this disclosure provides methods for treating neurodegenerative diseases or conditions associated with axonal degeneration.

[0495] In some embodiments, one or more compounds and / or compositions described herein may be used, for example, in medical practice. In some embodiments, one or more compounds and / or compositions described herein may be used, for example, to treat, prevent, or improve axonal degeneration (e.g., one or more of its characteristics or properties). In some embodiments, one or more compounds and / or compositions described herein may be used, for example, to inhibit axonal degeneration, including axonal degeneration caused by a reduction or depletion of NAD+. In some embodiments, one or more compounds and / or compositions described herein may be used, for example, to prevent axonal degeneration distal to axonal injury.

[0496] In some embodiments, one or more compounds and / or compositions as described herein can be used, for example, as a method to inhibit the degradation of neurons or portions thereof in the peripheral nervous system. In some embodiments, one or more compounds and / or compositions as described herein can be used, for example, as a method to inhibit or prevent degeneration of the central nervous system (neurons) or portions thereof. In some embodiments, one or more compounds or compositions as described herein are characterized by alleviating one or more symptoms or features of neurodegeneration when administered to a subject population. For example, in some embodiments, the associated symptoms or features may be selected from the degree, rate, and / or duration of neuronal damage.

[0497] In some embodiments, this disclosure provides compounds that can be used, for example, as analytical tools, probes in bioassays, or therapeutic agents according to this disclosure. The compounds provided in this disclosure can also be used to study SARM1 activity in biological and pathological phenomena, and to evaluate novel SARM1 activity inhibitors in vitro or in vivo. In some embodiments, this disclosure provides assays for identifying and / or characterizing the compounds and / or compositions provided herein. In some embodiments, the provided assays utilize specific reagents and / or systems (e.g., certain carrier constructs and / or peptides) that can be used to determine SARM1 activity. For example, in some embodiments, the provided assays may utilize: for example, SAM-TIR, wherein the SARM1 N-terminal self-repressive domain is missing; and / or one and / or multiple labeled forms of TIR domains.

[0498] In some embodiments, one or more compounds and / or compositions as described herein can be used, for example, as a method to inhibit neuronal degradation originating from a subject. In some embodiments, one or more compounds and / or compositions as described herein can be used to inhibit degeneration of neurons or portions thereof cultured in vitro. In some embodiments, one or more compounds and / or compositions as described herein can be used as stabilizers to promote neuronal survival in vitro.

[0499] In some embodiments, one or more compounds and / or compositions as described herein may be used, for example, to affect biomarkers associated with neurodegeneration. In some embodiments, changes in biomarkers may be detected systemically or using samples from a subject's CSF, plasma, serum, and / or tissues. In some embodiments, one or more compounds and / or compositions may be used to affect changes in the concentrations of NF-L and / or NF-H contained in a subject's CSF. In some embodiments, one or more compounds and / or compositions as described herein may affect constitutive NAD and / or cADPR levels in neurons and / or axons.

[0500] In some implementations, one or more biomarkers of neurodegeneration include: NF-L concentration from one or more of the following: obtained from CSF, blood, and plasma samples of the subject; NF-H concentration from one or more of the following: obtained from CSF, blood, and plasma samples of the subject; ubiquitin C-terminal hydrolase L1 (UCH-L1) concentration from one or more of the following: obtained from CSF, blood, and plasma samples of the subject; α-synuclein concentration from one or more of the following: obtained from CSF, blood, and plasma samples of the subject; constitutive NAD+ levels in neurons and / or axons of the subject; constitutive cADPR levels in neurons and / or axons of the subject; and albumin, amyloid-β (Aβ)38, Aβ40, Aβ42, GFAP, hFABP, MCP-1, neurogranulin, NSE, sAPPα, sAPPβ, sTREM, etc. 2. Phosphorus-tau or total-tau levels: obtained from CSF samples, blood samples, plasma samples, skin biopsy samples, nerve biopsy samples, and brain biopsy samples from the subject; and levels of one or more of the following: CCL2, CCL7, CCL12, colony-stimulating factor (CSF)1, or interleukin (IL)6: obtained from cerebrospinal fluid (CSF) samples, blood samples, plasma samples, skin biopsy samples, nerve biopsy samples, and brain biopsy samples from the subject.

[0501] In some embodiments, one or more compounds and / or compositions as described herein can affect the levels of one or more neurodegeneration-related proteins in a subject. These proteins include, but are not limited to, albumin, amyloid-β (Aβ)38, Aβ40, Aβ42, GFAP, hFABP, MCP-1, neurogranulin, NSE, sAPPα, sAPPβ, sTREM 2, phosphorus-tau, and / or total-tau. In some embodiments, one or more compounds and / or compositions as described herein can affect alterations in cytokines and / or chemokines, including but not limited to Ccl2, Ccl7, Ccl12, Csf1, and / or Il6.

[0502] Diseases, Disorders and Illnesses

[0503] In some embodiments, the compounds and / or compositions described herein may be administered to a subject suffering from one or more diseases, disorders, or conditions. In some embodiments, the one or more diseases, disorders, or conditions are mediated by SARM1.

[0504] In some implementations, neurodegenerative diseases or conditions include acute or chronic diseases or conditions of the peripheral nervous system (PNS), acute or chronic diseases or conditions of the central nervous system (CNS), or diseases associated with neurodegeneration.

[0505] In some embodiments, the neurodegenerative disease or condition includes an acute illness or condition of PNS. In some embodiments, the acute illness or condition of PNS is the result of mechanical injury, thermal injury, or injury from a chemical agent or chemotherapy. In some embodiments, mechanical injury includes compression or nerve entrapment injury or pressure injury. In some embodiments, compression or nerve entrapment injury includes carpal tunnel syndrome, direct trauma, penetrating injury, contusion, fracture, or dislocation of bone. In some embodiments, pressure injury includes pressure involving superficial nerves, pressure from tumors, or increased intraocular pressure. In some embodiments, the chemical agent or chemotherapy includes a cytotoxic anticancer agent, thalidomide, epochyam, taxane, vinca alkaloids, proteasome inhibitors, platinum-based drugs, or olistatin. In some embodiments, epochyam is ixabepilone. In some embodiments, taxane is paclitaxel or docetaxel. In some embodiments, the vinca alkaloid is vinblastine, vinorelbine, vincristine, or vindesine. In some embodiments, the proteasome inhibitor is bortezomib. In some embodiments, the platinum-based drug is cisplatin, oxaliplatin, or carboplatin. In some embodiments, olistatin is conjugated monomethyl olistatin E.

[0506] In some implementations, neurodegenerative diseases or conditions include chronic diseases or conditions of PNS. In some implementations, chronic diseases or conditions of PNS include systemic conditions, pain disorders, or metabolic diseases or conditions.

[0507] In some implementations, chronic diseases or conditions of PNS include hereditary neuropathy, Charcot-Marie-Tooth disease, hereditary sensory and autonomic neuropathy (HSAN), chronic inflammatory demyelinating polyneuropathy (CIDP), idiopathic neuropathy, or other peripheral neuropathy.

[0508] In some implementation schemes, systemic diseases include diabetes, uremia, AIDS, leprosy, nutritional deficiencies, atherosclerosis, enteric neuropathy, axonopathy, G. Barth syndrome, severe acute motor axononeuropathy (AMAN), systemic lupus erythematosus, scleroderma, sarcoidosis, rheumatoid arthritis, or polyarteritis nodosa.

[0509] In some implementations, pain disorders include chronic pain, fibromyalgia, spinal pain, carpal tunnel syndrome, cancer pain, arthritis, sciatica, headache, surgical pain, muscle spasm, back pain, visceral pain, injury pain, toothache, neurogenic pain, neuropathic pain, neuritis, nerve damage, herpes zoster, herniated disc, ligament tear, or diabetes.

[0510] In some implementations, metabolic diseases or conditions include diabetes, hypoglycemia, uremia, hypothyroidism, liver failure, polycythemia, amyloidosis, acromegaly, porphyria, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), lipid / glycolipid metabolism disorders, nutritional deficiencies, vitamin deficiencies, or mitochondrial diseases.

[0511] In some implementations, neurodegenerative diseases or conditions include acute diseases or conditions of the CNS. In some implementations, acute diseases or conditions of the CNS include ischemia, traumatic CNS injury, injury from chemical agents, thermal injury, or viral encephalitis.

[0512] In some implementations, ischemia includes cerebral ischemia, hypoxic demyelination, ischemic demyelination, ischemic optic neuropathy, or non-arteritis anterior ischemic optic neuropathy.

[0513] In some implementations, traumatic CNS injury includes spinal cord injury, TBI, mechanical injury to the head and / or spine, traumatic injury to the head and / or spine, blunt trauma, closed head injury, open head injury, exposure to shock and / or explosive forces, penetrating injury to the CNS, increased intraocular pressure, or injury from forces that cause axonal deformation, stretching, crushing, or sudden rotation.

[0514] In some implementations, viral encephalitis includes enterovirus encephalitis, arbovirus encephalitis, herpes simplex virus (HSV) encephalitis, West Nile virus encephalitis, La Crosse encephalitis, Bunyavirus encephalitis, pediatric viral encephalitis, or HIV encephalopathy (HIV-related dementia).

[0515] In some implementations, neurodegenerative diseases or conditions include chronic diseases or conditions of the CNS.

[0516] In some implementation schemes, chronic diseases or conditions of the CNS include Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS, Lou Griggs disease), multiple sclerosis (MS), Huntington's disease (HD), senile dementia, Pick's disease, Gaucher's disease, Hurler syndrome, progressive multifocal leukoencephalopathy, Alexander's disease, congenital hypomyelination, encephalomyelitis, acute disseminated encephalomyelitis, central pontine myelolysis, and osmotic hyponatremia. Hyponatremia, Tay-Sachs disease, motor neuron disease, ataxia, spinal muscular atrophy (SMA), Niemann-Pick disease, acute hemorrhagic leukoencephalitis, trigeminal neuralgia, Bell's palsy, cerebral ischemia, multiple system atrophy, Pelizaeus Merzbacher disease, periventricular leukomalacia, hereditary ataxia, noise-induced hearing loss, congenital hearing loss, age-related hearing loss, Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, Lewy body dementia, frontotemporal dementia Dementia, amyloidosis, diabetic neuropathy, globoid cell leukodystrophy (Krabbe's disease), Bassen-Kornzweig syndrome, transverse myelitis, motor neuron disease, spinocerebellar ataxia, pre-eclampsia, hereditary spastic paraplegia, spastic mild paraplegia.Paraparesis, familial spastic paraplegia, French settlement disease, Strumpell-Lorrain disease, non-alcoholic steatohepatitis (NASH), adrenomyeloneuropathy, progressive supranuclear palsy (PSP), Friedrich's ataxia, or spinal cord injury.

[0517] In some implementations, chronic diseases or conditions of the CNS include optic nerve disorders, traumatic CNS damage, or metabolic diseases or conditions.

[0518] In some implementations, optic nerve diseases include acute optic neuropathy, hereditary or idiopathic retinal diseases, Leber congenital amaurosis (LCA), Leber hereditary optic neuropathy (LHON), primary open-angle glaucoma (POAG), acute angle-closure glaucoma (AACG), autosomal dominant optic atrophy, retinal ganglion degeneration, retinitis pigmentosa, outer retinal neuropathy, optic nerve neuritis, optic nerve degeneration associated with multiple sclerosis, Kjer's optic neuropathy, ischemic optic neuropathy, vitamin B12 deficiency, folic acid (vitamin B9) deficiency, isolated vitamin E deficiency syndrome, non-arteritis anterior ischemic optic neuropathy, exposure to ethambutol, or exposure to cyanide.

[0519] In some implementations, traumatic CNS injury includes traumatic brain injury, spinal cord injury, traumatic axonal injury, or chronic traumatic encephalopathy (CTE).

[0520] In some implementations, metabolic diseases or conditions include diabetes, hypoglycemia, Barco syndrome, uremia, hypothyroidism, liver failure, polycythemia, amyloidosis, acromegaly, porphyria, lipid / glycolipid metabolism disorders, nutritional / vitamin deficiencies, and mitochondrial diseases.

[0521] In some implementations, neurodegenerative diseases or conditions include those related to neurodegeneration. In some implementations, neurodegenerative diseases or conditions are caused by clotting problems, inflammation, obesity, aging, stress, cancer, or diabetes.

[0522] In some implementations, the condition is acute peripheral neuropathy. Chemotherapy-induced peripheral neuropathy (CIPN) is an example of acute peripheral neuropathy. CIPN can be associated with a variety of drugs, such as, but not limited to, thalidomide, ephemeralmycin (e.g., ixaprazole), taxanes (e.g., paclitaxel and docetaxel), vinblastine alkaloids (e.g., vinblastine, vinorelbine, vincristine, and vindesine), proteasome inhibitors (e.g., bortezomib), and platinum-based drugs (e.g., cisplatin, oxaliplatin, and carboplatin).

[0523] In some embodiments, one or more compounds and / or compositions as described herein are used, for example, to treat one or more neurodegenerative diseases, disorders, or conditions selected from neuropathy or axonopathy. In some embodiments, one or more compounds and / or compositions as described herein can be used, for example, to treat neuropathy or axonopathy associated with axonodegeneration. In some embodiments, the neuropathy associated with axonodegeneration is a hereditary or congenital neuropathy or axonopathy. In some embodiments, the neuropathy associated with axonodegeneration is caused by a de novo mutation or somatic mutation. In some embodiments, the neuropathy associated with axonodegeneration is selected from the list included herein. In some embodiments, the neuropathy or axonopathy is associated with axonodegeneration, including but not limited to Parkinson's disease, non-Parkinson's disease, Alzheimer's disease, herpes infection, diabetes, amyotrophic lateral sclerosis, demyelinating diseases, ischemic or stroke, chemical injury, thermal injury, and AIDS.

[0524] In some embodiments, one or more compounds or compositions as described herein are characterized by alleviating one or more symptoms or features of neurodegeneration when administered to a subject population. For example, in some embodiments, the associated symptoms or features may be selected from the degree, rate, and / or timing of neuronal damage. In some embodiments, neuronal damage may be or include axonal degradation, synaptic loss, dendritic loss, synaptic density loss, dendritic arborization loss, axonal branching loss, neuronal density loss, myelination loss, neuronal cell body loss, synaptic enhancement loss, action potential enhancement loss, cytoskeleton stability loss, axonal transport loss, ion channel synthesis and turnover loss, neurotransmitter synthesis loss, neurotransmitter release and reuptake capacity loss, axonal potential propagation loss, neuronal hyperexcitability, and / or neuronal hypoexcitability. In some embodiments, neuronal damage is characterized by the inability to maintain an appropriate resting neuronal membrane potential. In some embodiments, neuronal damage is characterized by the presence of inclusion bodies, plaques, and / or neurofibrillary tangles. In some embodiments, neuronal damage is characterized by the presence of stress granules. In some embodiments, neuronal destruction is characterized by intracellular activation of one or more members of the cysteine-aspartic protease (cystase) family. In some embodiments, neuronal destruction is characterized by neurons undergoing programmed cell death (e.g., apoptosis, pyroptosis, ferroptosis, and / or necrosis) and / or inflammation.

[0525] In some implementations, neurodegeneration or neurological diseases or conditions are associated with axonal degeneration, axonal injury, axonopathy, demyelinating diseases, central pontine myelination, neurological injury diseases or conditions, metabolic diseases, mitochondrial diseases, metabolic axonal degeneration, and axonal injury caused by leukoencephalopathy or leukodystrophy. In some implementations, neurodegeneration or neurological disease or condition is selected from spinal cord injury, stroke, multiple sclerosis, progressive multifocal leukoencephalopathy, congenital hypomyelination, encephalomyelitis, acute disseminated encephalomyelitis, central pontine myelination, osmotic hyponatremia, hypoxic demyelination, ischemic demyelination, adrenoleukodystrophy, Alexander disease, Niemann-Pick II disease, Peyreke-Merck II disease, periventricular leukomalacia, glomerular leukodystrophy (Clapper's disease), Wallerian degeneration, optic neuritis, transverse myelitis, amyotrophic lateral sclerosis (ALS, Lou Gehrig's disease). Gehrig's disease, Huntington's disease, Alzheimer's disease, Parkinson's disease, Tay-Sachs disease, Gaucher disease, Heller syndrome, traumatic brain injury, radiation injury, neurological complications of chemotherapy (chemotherapy-induced neuropathy; CIPN), neuropathy, acute ischemic optic neuropathy, vitamin B12 12 Vitamin E deficiency, isolated vitamin E deficiency syndrome, Barco syndrome, glaucoma, Leber hereditary optic atrophy (neuropathy), Leber congenital amaurosis, neuromyelitis optica, metachromatic leukodystrophy, acute hemorrhagic leukoencephalitis, trigeminal neuralgia, Bell's palsy, cerebral ischemia, multiple system atrophy, traumatic glaucoma, tropical spastic paraplegia. Paraparesis), human T-lymphotropic virus 1 (HTLV-1)-associated myelopathy, West Nile virus encephalopathy, earth virus encephalitis, Bunyavirus encephalitis, pediatric viral encephalitis, essential tremor, Sharma-Tussan disease, motor neuron disease, SMA, HSAN, adrenocortical neuropathy, PSP, Friedrich's ataxia, hereditary ataxia, noise-induced hearing loss, congenital hearing loss, Lewy body dementia, frontotemporal dementia, amyloidosis, diabetic neuropathy, HIV neuropathy, enteric neuropathy and axonopathy, Göttingen-Bartholin's syndrome, AMAN, Creutzfeldt-Jakob disease, infectious spongiform encephalopathy, spinocerebellar ataxia, preeclampsia, hereditary spastic paraplegia, spastic mild paraplegia, familial spastic paraplegia, French residency disease, Strumpell-Lorrain disease, and NASH.

[0526] In some embodiments, this disclosure provides SARM1 activity inhibitors for treating neurodegenerative or neurological diseases or conditions involving axonal degeneration or axonopathy. This disclosure also provides methods for treating, preventing, or improving axonal degeneration, axonopathy, and neurodegenerative or neurological diseases or conditions involving axonal degeneration using SARM1 activity inhibitors.

[0527] In some embodiments, this disclosure provides methods for treating neurodegeneration or neurological diseases or conditions related to axonal degeneration, axonal injury, axonopathy, demyelinating diseases, central pontine myelination, nerve injury diseases or conditions, metabolic diseases, mitochondrial diseases, metabolic axonal degeneration, and axonal injury caused by leukodystrophy or leukodystrophy.

[0528] In some implementations, neuropathy and axonopathy include any disease or condition involving neurons and / or supporting cells, such as glial cells, myocytes, or fibroblasts, particularly those involving axonal injury. Axonal injury can be caused by traumatic injury or non-mechanical injury resulting from disease, condition, or exposure to toxic molecules or drugs. The result of such injury can be axonal degeneration or dysfunction, as well as loss of functional neuronal activity. A wide range of neurological diseases and conditions can produce or be associated with such axonal injury. Such neuropathy can include peripheral neuropathy, central neuropathy, and combinations thereof. Furthermore, peripheral neurological manifestations can arise from diseases primarily affecting the central nervous system, and central nervous system manifestations can arise from diseases that are primarily peripheral or systemic.

[0529] In some implementations, peripheral neuropathy may involve damage to the peripheral nerves and / or may be caused by neurological disorders or be a result of systemic diseases. Some such diseases may include diabetes, uremia, infectious diseases such as AIDS or leprosy, nutritional deficiencies, vascular or collagen disorders (such as atherosclerosis), and autoimmune diseases (such as systemic lupus erythematosus, scleroderma, sarcoidosis, rheumatoid arthritis, and polyarteritis nodosa). In some implementations, peripheral nerve degeneration is caused by traumatic (mechanical) injury to the nerves as well as chemical or thermal injury. These conditions of peripheral nerve injury include compression or contusion injuries, such as glaucoma, carpal tunnel syndrome, direct trauma, penetrating injuries, contusions, fractures, or dislocations; compression involving superficial nerves (ulnar, radial, or peroneal nerves), which may be caused by prolonged use of crutches or staying in one position for too long or by tumors; intraneural hemorrhage; ischemia; exposure to cold or radiation or certain drugs or toxic substances such as herbicides or pesticides. Specifically, nerve damage can be caused by chemical damage resulting from cytotoxic anticancer agents (such as paclitaxel, cisplatin, proteasome inhibitors, or vinca alkaloids like vincristine). Typical symptoms of this type of peripheral neuropathy include weakness, numbness, paresthesia (abnormal sensations such as burning, itching, tingling, or numbness), and pain in the arms, hands, legs, and / or feet. In some implementations, the neuropathy is associated with mitochondrial dysfunction. This type of neuropathy can manifest as reduced energy levels, specifically decreased NAD and ATP levels.

[0530] In some implementations, peripheral neuropathy is a metabolic and endocrine neuropathy, encompassing a broad spectrum of peripheral neuropathy associated with systemic diseases of metabolic origin. These diseases include, for example, diabetes mellitus, hypoglycemia, uremia, hypothyroidism, liver failure, polycythemia vera, amyloidosis, acromegaly, porphyria, lipid / glycolipid metabolism disorders, nutritional / vitamin deficiencies, and mitochondrial disorders. A common hallmark of these diseases is involvement of the peripheral nerves through alterations in the structure or function of myelin and axons due to metabolic pathway dysregulation.

[0531] In some implementations, neuropathy includes optic neuropathy such as glaucoma; retinal ganglion degeneration, such as those associated with retinitis pigmentosa and external retinal neuropathy; optic neuritis and / or degeneration, including those associated with multiple sclerosis; traumatic injury to the optic nerve, which may include, for example, injury during tumor resection; hereditary optic neuropathy, such as Kjer's disease and Leber's hereditary optic neuropathy; ischemic optic neuropathy, such as those secondary to giant cell arteritis; metabolic optic neuropathy, such as neurodegenerative diseases, including the aforementioned Leber neuropathy, nutritional deficiencies such as vitamin B12 or folic acid deficiency, and toxicities such as ethambutol or cyanide toxicity; neuropathy caused by adverse drug reactions; and neuropathy caused by vitamin deficiencies. Ischemic optic neuropathy also includes non-arteritis anterior ischemic optic neuropathy.

[0532] In some implementations, neurodegenerative diseases associated with neuropathy or axonopathy in the central nervous system include a variety of conditions. These include those involving progressive dementia, such as Alzheimer's disease, senile dementia, Pick's disease, and Huntington's disease; central nervous system diseases affecting muscle function, such as Parkinson's disease, motor neuron disease, and progressive ataxia, such as amyotrophic lateral sclerosis; demyelinating diseases, such as multiple sclerosis; viral encephalitis, such as viral encephalitis caused by enteroviruses, arboviruses, and herpes simplex virus; and prion diseases. Mechanical injuries such as glaucoma or traumatic injuries to the head and spine can also cause nerve damage and degeneration in the brain and spinal cord. In addition, local ischemia and stroke, as well as conditions such as nutritional deficiencies and chemical toxicity (e.g., chemotherapeutic agents), can cause central nervous system neuropathy.

[0533] In some embodiments, this disclosure provides methods for treating neuropathies or axonopathy associated with axonal degeneration. In some such embodiments, the neuropathy or axonopathy associated with axonal degeneration can be any of many neuropathies or axonopathy, such as those that are hereditary or congenital, or associated with Parkinson's disease, Alzheimer's disease, herpes infections, diabetes, amyotrophic lateral sclerosis, demyelinating diseases, ischemic or stroke, chemical injuries, thermal injuries, and AIDS. Furthermore, neurodegenerative diseases not mentioned above, as well as subsets of the aforementioned diseases, can also be treated using the methods of this disclosure. Such subsets of diseases can include Parkinson's disease or non-Parkinson's disease or Alzheimer's disease.

[0534] Subjects

[0535] In some embodiments, the compounds and / or compositions described herein are administered to a subject who has or is susceptible to the diseases, disorders, or conditions described herein; in some embodiments, such diseases, disorders, or conditions are characterized by axonal degeneration, such as one of the symptoms mentioned herein.

[0536] In some embodiments, subjects who have been given the compound or composition as described herein exhibit one or more signs or symptoms associated with axonal degeneration; in other embodiments, subjects do not exhibit any signs or symptoms of neurodegeneration.

[0537] In some embodiments, the provided method includes administering a compound of formula I to a patient in need. In some such embodiments, the patient is at risk of developing a condition characterized by axonal degeneration. In some embodiments, the patient already has a condition characterized by axonal degeneration. In some embodiments, the patient has been diagnosed with a condition characterized by axonal degeneration.

[0538] In some embodiments, the provided method includes administering the composition described herein to a patient population in need. In some embodiments, the population consists of individuals engaged in activities with a high likelihood of traumatic neuronal injury. In some embodiments, the population consists of athletes engaged in contact sports or other high-risk activities.

[0539] In some implementations, the subject is at risk of developing a condition characterized by axonal degeneration. In some implementations, the subject is identified as being at risk of axonal degeneration, for example, based on the subject's genotype, diagnosis of a condition associated with axonal degeneration, and / or exposure to agents and / or conditions that induce axonal degeneration.

[0540] In some implementations, the patient is at risk of developing a neurodegenerative disease. In some implementations, the patient is middle-aged or older. In some implementations, the patient is known to have genetic risk factors for neurodegeneration. In some implementations, the patient has a family history of neurodegenerative disease. In some implementations, the patient expresses one or more copies of a known genetic risk factor for neurodegeneration. In some implementations, the patient was selected from a population with a high incidence of neurodegeneration. In some implementations, the patient has a hexanucleotide repeat amplification in open reading frame 72 of chromosome 9. In some implementations, the patient has one or more copies of the ApoE4 allele.

[0541] In some embodiments, the subject administering the compounds or compositions described herein may be or include subjects who have or are predisposed to neurodegenerative diseases, disorders, or conditions. In some embodiments, neurodegenerative diseases may be or include traumatic neuronal injury. In some embodiments, traumatic neuronal injury is blunt force trauma, closed head injury, open head injury, exposure to concussive and / or explosive forces, or penetrating injury to the brain cavity or nerve-innervated areas of the body. In some embodiments, traumatic neuronal injury is a force that causes axonal deformation, stretching, crushing, or sudden rotation.

[0542] In some implementations, subjects engage in activities identified as risk factors for neuronal degradation; for example, subjects engaged in contact sports or occupations have a high chance of traumatic neuronal injury.

[0543] For example, subjects may be patients who are receiving or have been prescribed chemotherapy for peripheral neuropathy. Examples of chemotherapeutic agents include, but are not limited to, thalidomide, epokines (e.g., ixaprazole), taxanes (e.g., paclitaxel and docetaxel), vinblastine alkaloids (e.g., vinblastine, vinorelbine, vincristine, and vindesine), proteasome inhibitors (e.g., bortezomib), and platinum-based drugs (e.g., cisplatin, oxaliplatin, and carboplatin).

[0544] In some embodiments, the provided method includes administering the composition described herein to a patient or patient population based on the presence or absence of one or more biomarkers. In some embodiments, the provided method further includes monitoring the levels of biomarkers in the patient or patient population and adjusting the dosing regimen accordingly.

[0545] Dosage

[0546] Those skilled in the art will understand that, in some embodiments, the exact amount of a particular compound included in and / or delivered by administration of the pharmaceutical compositions or regimens described herein may be chosen by a medical practitioner and may vary for different subjects, for example, when considering one or more of the subject's species, age and general condition and / or the characteristics of the particular compound or composition, its administration method, etc. Alternatively, in some embodiments, the amount of a particular compound included in and / or delivered by administration of the pharmaceutical compositions or regimens described herein may be standardized across the relevant patient population (e.g., all patients, a specific age or disease stage, or all patients expressing a specific biomarker, etc.).

[0547] The compounds or compositions provided in this disclosure are preferably formulated in unit dosage form for ease of administration and uniform dosage. As used herein, "unit dosage form" refers to a physically discrete unit of the agent suitable for the patient to be treated. However, it should be understood that the total daily dosage of the compounds or compositions provided in this disclosure will be determined by the attending physician within reasonable medical judgment. The specific effective dose level for any particular patient or organism will depend on a variety of factors, including the condition being treated and its severity; the individual patient's clinical condition; the cause of the disease; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health condition, sex, and diet; the time of administration, the site of delivery, the route of administration, and the rate of excretion of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical field. The effective amount of the compound to be administered will be determined by these considerations and is the minimum amount required to inhibit SARM1 activity as needed to prevent or treat undesirable diseases or conditions (e.g., neurodegeneration or traumatic neurological injury).

[0548] The pharmaceutically acceptable compositions of this disclosure can be administered to humans and other animals orally, rectally, intravenously, parenterally, intracerebrospinally, vaginally, intraperitoneally, topically (e.g., by powder, ointment, or drops), sublingually, as oral or nasal sprays, etc., depending on the severity of the disease, condition, or infection being treated. In some embodiments, the daily dose is administered as a single daily dose or in divided doses two to six times daily, or in a sustained-release form. This dosing regimen can be adjusted to provide the optimal therapeutic response. The compounds can be administered 1 to 4 times daily, preferably once or twice daily.

[0549] In some embodiments, the compositions of this disclosure can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or through an implanted reservoir. As used herein, the term "parenterical" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intradermal, intraocular, intralesional, and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally, or intravenously.

[0550] In some embodiments, the pharmaceutically acceptable compositions of this disclosure can also be applied topically, particularly when the treatment target includes areas or organs easily accessible for topical application, including eye, skin, or lower intestinal diseases. Suitable topical formulations are readily prepared for each of these areas or organs.

[0551] Most preferably, the pharmaceutically acceptable compositions of this disclosure are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions of this disclosure are administered without food. In other embodiments, the pharmaceutically acceptable compositions of this disclosure are administered with food.

[0552] As part of a multiple-dosing regimen, the additional agents may be administered separately from the provided compound or its composition. Alternatively, the agents may be part of a single dosage form, mixed together with the provided compound in a single composition. If administered as part of a multiple-dosing regimen, the two active agents may be administered simultaneously, sequentially, or over a period of time, typically within 5 hours of each other.

[0553] It should also be understood that the specific dosage and treatment regimen for any particular patient can depend on a variety of factors, including the activity of the specific compound used, age, weight, general health condition, sex, diet, timing of administration, excretion rate, drug combination, and the judgment of the treating physician and the severity of the specific disease being treated. In some embodiments, the amount of the disclosed compound in the composition will also depend on the specific compound in the composition.

[0554] In some embodiments, the SARM1 inhibitor described herein may be used in combination with one or more other therapies to treat an associated disease, disorder, or condition. In some embodiments, the dose of the SARM1 inhibitor is changed when used in combination therapy compared to when administered as a monotherapy; optionally or additionally, in some embodiments, the therapy administered in combination with the SARM1 inhibitor as described herein is administered according to a different regimen than when administered alone or in combination with one or more therapies other than SARM1 inhibitor. In some embodiments, the composition comprises an additional therapeutic agent that may act synergistically with the provided compound. In some embodiments, one or both therapies used in the combination regimen are administered at a lower level or at a lower frequency than when used as monotherapy.

[0555] In some embodiments, the compounds and / or compositions described herein are administered in combination with chemotherapeutic agents, including but not limited to alkylating agents, anthracyclines, taxanes, epoch-forming agents, histone deacetylase inhibitors, topoisomerase inhibitors, kinase inhibitors, nucleotide analogs, peptide antibiotics, platinum-based agents, retinoids, vinca alkaloids, and derivatives. In some embodiments, the compounds and / or compositions described herein are administered in combination with PARP inhibitors. Example

[0556] This teaching, including the descriptions provided in the embodiments, is not intended to limit the scope of any claim. Unless specifically presented in the past tense, inclusion in the embodiments is not intended to imply that an experiment was actually performed. The following non-limiting embodiments are provided to further illustrate this teaching. Based on this disclosure, those skilled in the art will understand that many changes can be made to the specific embodiments disclosed without departing from the spirit and scope of this teaching, and still obtaining the same or similar results.

[0557] method

[0558] Some of the methods and compositions described herein utilize laboratory techniques well known to those skilled in the art and can be found in laboratory manuals, such as Sambrook, J. et al., Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2001; Methods In Molecular Biology, ed. Richard, Humana Press, NJ, 1995; Spector, DL et al., Cells: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1998; and Harlow, E., Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1999. The administration method and dosage regimen of a drug can be determined based on standard pharmacological principles and using methods provided in standard references, such as Remington: The Science and Practice of Pharmacy (Alfonso R. Gennaro ed. 19th ed. 1995); Hardman, JG et al., Goodman & Gilman's The Pharmacological Basis of Therapeutics, 9th ed., McGraw-Hill, 1996; and Rowe, RC et al., Handbook of Pharmaceutical Excipients, 4th ed., Pharmaceutical Press, 2003.

[0559] Example 1: Synthesis of the compound

[0560] General Synthesis Method

[0561] The compounds and intermediates according to the invention can be obtained using synthetic methods known to those skilled in the art and described in the organic synthesis literature. Preferably, the compounds are obtained in a manner similar to the preparation methods explained more fully below, particularly as described in the experimental section. In some cases, the order of the reaction steps can be varied. Variations of reaction methods known to those skilled in the art but not described in detail herein can also be used.

[0562] The general methods for preparing the compounds of the present invention will become apparent to those skilled in the art who study the following schemes. The starting materials can be prepared by methods described in the literature or herein, or by similar or analogous methods. Any functional groups in the starting materials or intermediates can be protected with conventional protecting groups. These protecting groups can be cleaved again at appropriate stages within the reaction sequence using methods well known to those skilled in the art.

[0563] Optimal reaction conditions and reaction times can vary depending on the specific reactants used. Unless otherwise specified, those skilled in the art can readily select solvents, temperatures, pressures, and other reaction conditions. Specific methods are provided in the Synthesis Examples section. Intermediates and products can be purified by silica gel chromatography, recrystallization, and / or reversed-phase HPLC (RHPLC). Discrete enantiomers can be obtained by resolving racemic products using chiral HPLC. RHPLC purification methods in any case use a 0-100% aqueous solution of acetonitrile containing 0.1% formic acid, 0.1-0.01% TFA, 10 mM ammonium bicarbonate, or 0.2% ammonium hydroxide, and employ one of the following columns:

[0564] Waters Xbridge C18 10μm 30x100 mm column

[0565] Waters Sunfire C18 10μm 30x100 mm column

[0566] Waters Xbridge C18 3.5μm 50x4.6 mm column

[0567] HALO C18 2.7μm 30x4.6 mm column

[0568] Waters Sunfire C18 3.5μm 50x4.6 mm column

[0569] Synthesis of exemplary compounds

[0570] Method A: Synthesis of compound I-1-a

[0571]

[0572] To a solution of R-2 (1.87 g, 6.24 mmol), R-1 (1.00 g, 6.24 mmol), and DIPEA (2.2 mL, 12.5 mmol) in anhydrous DMF (10 mL), a solution of propionic anhydride in EtOAc (50%, 5.6 mL, 9.36 mmol) was added. The reaction mixture was purged with N2 (g), sealed, and stirred at room temperature for 1 hour. A saturated aqueous solution of NaHCO3 (10 mL) and water (10 mL) were added, and the mixture was extracted with CH2Cl2 (3 x 20 mL). The combined organic layers (MgSO4) were dried, filtered, and concentrated under vacuum. The product was ground in CH2Cl2 (10 mL), and the solid was collected by vacuum filtration and washed with CH2Cl2 (2 x 5 mL) to give Int-1 (490 mg, 17%).

[0573] Int-1 (1.14 g, 1.55 mmol) was dissolved in CH2Cl2 (5 mL) at room temperature. Trifluoroacetic acid (2.0 mL, 26.1 mmol) was added, and the reaction mixture was stirred for 16 hours. The reaction mixture was concentrated under vacuum, and the crude product was dissolved in CH2Cl2 (20 mL), followed by the addition of saturated NaHCO3 aqueous solution (10 mL) and water (10 mL). The organic layer was separated, and the aqueous layer was extracted with CH2Cl2 (2 x 20 mL). The organic layers were combined, dried (MgSO4), filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by preparative HPLC to obtain the free amine, which was then suspended in MeOH (2 mL). Chloroacetaldehyde aqueous solution (50% wt, 47 μL, 0.37 mmol) was added to the suspension, and the mixture was stirred for 4 hours, followed by treatment with NaCNBH3 (23 mg, 0.37 mmol). The mixture was stirred for 16 h, then treated with water (2 mL), extracted with CH2Cl2 (3 x 2 mL), dried with MgSO4, filtered, and concentrated under vacuum. The crude product (SiO2, 0-5% MeOH in CH2Cl2 solution) was purified by rapid chromatography to give Int-2 (40 mg, 29%).

[0574] Sodium hydride (60% dispersion in mineral oil, 5.8 mg, 0.14 mmol) was added to a solution of Int-2 (73%, 40 mg, 0.072 mmol) in anhydrous DMF (2 mL) at room temperature. The reaction mixture was stirred for 1 hour under a N2 (g) atmosphere. The reaction was quenched with water (2 mL), and the mixture was extracted with CH2Cl2 (3 x 2 mL). The combined organic layers (MgSO4) were dried, filtered, and concentrated under vacuum. The crude product was purified by preparative HPLC to give compound I-1-a (17 mg, 63%).

[0575] The following compounds, I-1-b, I-3-a, and I-38, were prepared in a similar manner from suitable amines and acid reagents.

[0576] Method B: Synthesis of compound I-2-a

[0577]

[0578] Sodium cyanoborohydride (19 mg, 0.31 mmol) was added to a solution of compound I-1-a (88%, 86 mg, 0.21 mmol) and formaldehyde (12 mg, 0.41 mmol) in methanol (4 mL) at room temperature, and the reaction mixture was stirred for 22 hours. The reaction was quenched with water (4 mL), and the mixture was extracted with v (4 x 10 mL). The combined organic layers (MgSO4) were dried, filtered, and concentrated under vacuum. The crude product was purified by preparative HPLC to give compound I-2-a (43 mg, 54%).

[0579] Method C: Synthesis of compound I-4

[0580]

[0581] LiHMDS (1 mol / L, 81 mL, 81 mmol) was slowly added to a mixture of R-3 (15 g, 81 mmol) and THF (100 mL) at -60 °C over 1 hour. 1-(bromomethyl)-4-chlorobenzene (16 g, 81 mmol) was added to THF (100 mL), and the mixture was stirred at -60 °C for 1 hour. The mixture was poured into water (100 mL) and extracted with ethyl acetate (100 mL x 2). The combined organic layers were dried, concentrated under vacuum to give a crude product, which was purified by column chromatography (SiO2, CH2Cl2:MeOH = 100:1) to give pyrrolidone (8.0 g, 26 mmol), which was dissolved in THF / H2O (4 / 1, 80 mL). H2O2 (10 mL) was slowly added to this solution at 0 °C. LiOH-H2O (3.3 g, 78 mmol) was added, and the mixture was stirred at room temperature for 2 hours. It was then treated with saturated NaHSO3 (15 mL), extracted with ethyl acetate (100 mL x 2), dried, and concentrated under vacuum to give an acid (7.8 g, 24.0 mmol). This acid was treated with TFA (2 mL) in CH2Cl2 (5 mL) and stirred at room temperature for 2 hours. The reaction mixture was then concentrated under vacuum to give Int-3 (5.5 g, quantified).

[0582] A mixture of Int-3 (5.5 g, 24 mmol), 3,5-dibromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-1,2,4-triazole (12.8 g, 36 mmol), and K₂CO₃ (9.9 g, 72 mmol) in H₂O / 1,4-dioxane (1 / 10, 50 mL) was stirred at 110 °C for 32 h. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (200 mL x 2). The combined organic layers were dried and concentrated under vacuum. The residue was purified by rapid chromatography (SiO₂, DCM:MeOH = 20:1) to give an amine (7.0 g, 14 mmol), which was dissolved in DMA (20 mL). Propionic anhydride (13.4 g, 42 mmol) and DIPEA (3.6 g, 28 mmol) were added to the mixture, and the mixture was stirred at 40 °C for 2 h. Add water (60 mL), extract the mixture with ethyl acetate (200 mL x 2), dry, concentrate under vacuum to obtain crude product, and purify by rapid chromatography (SiO2, DCM:MeOH = 30:1) to obtain Int-4 (5.5 g, 81%).

[0583] A mixture of Int-4 (5.5 g, 11 mmol), pyridin-4-ylboronic acid (2.1 g, 17 mmol), Pd(dppf)Cl2 (804 mg, 1.1 mmol), and K2CO3 (4.6 g, 33 mmol) in H2O / 1,4-dioxane (1 / 10, 50 mL) was stirred at 110 °C for 16 h. The reaction mixture was treated with water (50 mL) and extracted with ethyl acetate (100 mL x 2). The combined organic layers were dried, concentrated under vacuum, and the crude product was purified by column chromatography (SiO2, DCM:MeOH = 20:1) to give pyridine (4.6 g, 9.5 mmol), which was treated with CH2Cl2 (5 mL) and TFA (5 mL). The mixture was stirred for 2 h and then concentrated under vacuum to give a crude product, which was purified by rapid chromatography (SiO2, DCM:MeOH = 20:1) to give the title compound (3.2 g, 95%).

[0584] The following compounds, I-5 and I-6, are prepared in a similar manner from suitable amine and acid reagents.

[0585] Method D: Synthesis of Compound I-7

[0586]

[0587] A suspension of R-4 (5.0 g, 14.0 mmol), pyrrolidone-2-one (3.2 mL, 42.0 mmol), tripotassium phosphate (5.94 g, 28.0 mmol), cuprous iodide (1+) (0.27 g, 1.40 mmol), and 1,10-phenanthroline (0.76 g, 4.20 mmol) in anhydrous DMF (100 mL) was degassed for 5 minutes with a stream of N2 (g). The reaction mixture was stirred at 100 °C under an N2 (g) atmosphere for 5 hours. The reaction mixture was then concentrated under vacuum. The product was partitioned between EtOAc and brine, and the layers were separated. The aqueous layer was re-extracted with EtOAc. The combined organic layers (MgSO4) were dried and concentrated under vacuum. The crude product (SiO2, 0%-100% EtOAc in heptane solution) was purified by flash column chromatography to give pyrrolidone (1.81 g, 5.01 mmol), which was dissolved in 1,4-dioxane (15 mL). 4-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)pyridine (1.34 g, 6.51 mmol) was added, followed by 2M potassium carbonate aqueous solution (7.5 mL, 15.0 mmol). The reaction mixture was degassed with N2(g) for 5 min. 1,1'-bis(diphenylphosphino)ferrocene]palladium(II) chloride (367 mg, 0.500 mmol) was added, and the reaction mixture was stirred at 100 °C under N2(g) atmosphere for 5 h. The reaction mixture was diluted with EtOAc and brine. The layers were separated, and the aqueous layer was extracted with EtOAc. The combined organic layers were concentrated under vacuum. The crude product was purified by flash column chromatography using a gradient of 0%–100% EtOAc in heptane solution. The residue was suspended in 75 mL of hot water at 40 °C and extracted with 100 mL of EtOAc. The organic layer was washed with 75 mL of hot water and then with 50 mL of brine. The organic layer (MgSO4) was dried, filtered, and concentrated under vacuum in a water bath set at 60 °C to give Int-5 (1430 mg, 71%).

[0588] Int-5 (90%, 200 mg, 0.501 mmol) was dissolved in anhydrous THF (3 mL). The reaction mixture was cooled to -78 °C under a N2 (g) atmosphere, and a THF solution of LiHMDS (1 M, 28 μL, 0.25 mmol) was added dropwise. The reaction mixture was stirred at -78 °C for 15 min, and then 1-(bromomethyl)-4-fluorobenzene (69 μL, 0.55 mmol) was added dropwise to anhydrous THF (1 mL). The reaction mixture was stirred at -78 °C for 1 h, then quenched with water (5 mL), diluted with brine (30 mL), and extracted with EtOAc (3 x 40 mL). The combined organic layers (MgSO4) were dried, filtered, and concentrated under vacuum. The crude product was purified by flash column chromatography using a gradient of 0%–100% EtOAc in heptane to give a benzylated compound (150 mg, 0.23 mmol), which was dissolved in DCM (4 mL). Add TFA (0.5 mL) to the mixture and allow it to stand at room temperature for 20 hours. Quench the reaction mixture with a saturated aqueous solution of NaHCO3, and collect the solid by vacuum filtration. Wash the solid with water and then grind it in methanol (2 mL) to give the title compound (14 mg, 17%).

[0589] The following compounds were prepared in a similar manner from suitable lactams and alkylating agents: I-8, I-9, I-10, I-11, I-12, I-13, I-18, I-26, I-39, I-40, I-42-45, I-54, I-61, I-63, I-95, I-97, I-99, I-101-103, I-111, and I-115.

[0590] Method E: Synthesis of Compound I-15

[0591]

[0592] R-5 (7.10 g, 31.4 mmol) was suspended in anhydrous THF (100 mL) and cooled to 0 °C. Sodium hydride (60% mineral oil solution, 1.26 g, 31.4 mmol) was added stepwise to the reaction mixture. The reaction mixture was stirred at room temperature under a N2(g) atmosphere for 10 min. [2-(chloromethoxy)ethyl](trimethyl)silane (6.1 mL, 34.6 mmol) was added, and the reaction mixture was stirred under a N2(g) atmosphere for 18 h. The reaction mixture was quenched with water and extracted with EtOAc (20 mL). The organic layer was washed with a saturated NaHCO3 aqueous solution (20 mL) and then concentrated under vacuum. The crude product was purified by flash column chromatography with a gradient of 0-10% EtOAc in heptane to give SEM-protected pyrazole (9.10 g, 80%).

[0593] SEM-protected pyrazole (2.00 g, 5.45 mmol), potassium carbonate (2.26 g, 16.3 mmol), bis(diphenylphosphino)ferrocene]dichloropalladium(II) (400 mg, 0.55 mmol), and 4-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)pyridine (1.12 g, 5.45 mmol) were placed in a N2(g) atmosphere. Water (5 mL) and 1,4-dioxane (15 mL) were added to the reaction mixture, and the mixture was purged with N2(g). The reaction mixture was stirred at 100 °C for 45 min. The reaction mixture was cooled to room temperature, and EtOAc (150 mL) and brine (100 mL) were partitioned. The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 100 mL). The combined organic layers (MgSO4) were dried, filtered, and concentrated under vacuum. The crude product was purified by a flash column chromatography with a gradient of 0-100% EtOAc in heptane solution to obtain Int-6 (3.07 g, 39%).

[0594] Int-6 (3.00 g, 8.30 mmol), pyrrolidone-2-one (1.3 mL, 16.6 mmol), tripotassium phosphate (3.57 g, 16.6 mmol), BrettPhos Pd G1 methyl tert-butyl ether adduct (128 mg, 0.14 mmol), Brettphos (445 mg, 0.830 mmol), and Pd2(dba)3 (380 mg, 0.42 mmol) were combined and dissolved in anhydrous 1,4-dioxane (60 mL) under a nitrogen atmosphere. The reaction mixture was stirred at 110 °C for 18 hours. The reaction mixture was cooled to room temperature and concentrated under vacuum. The crude product was purified by flash column chromatography with a gradient of 0-100% EtOAc in heptane to give Int-7 (2.31 g, 76%).

[0595] Int-7 (400 mg, 1.12 mmol) was dissolved in anhydrous THF (7 mL), cooled to 78 °C, and then treated with a THF solution of LiHMDS (1 M, 1.3 mL, 1.34 mmol) with stirring for 10 min. 1-(bromomethyl)-4-chlorobenzene (229 mg, 1.12 mmol) was added dropwise to anhydrous THF (5 mL), and the reaction mixture was stirred at -78 °C for 2 h. The reaction mixture was quenched with water (30 mL) and brine (30 mL), and then extracted with EtOAc (3 x 50 mL). The combined organic layers (Na₂SO₄) were dried, filtered, and concentrated under vacuum. The crude product was purified by flash column chromatography using a gradient of 0%–100% EtOAc in heptane to give a benzylated compound (417 mg, 0.85 mmol), which was dissolved in DCM (6 mL). Trifluoroacetic acid (3.0 mL, 40.4 mmol) was added to the mixture, and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then concentrated under vacuum. The crude product was dissolved in an acetonitrile / water mixture (1:1, 2 mL). A saturated aqueous solution of ammonium hydroxide (1.0 mL) was added, and the mixture was sonicated for 2 minutes to obtain a white precipitate. The volatile substances were concentrated under vacuum, and the solid collected was vacuum filtered and washed with water. The solid was suspended in boiling acetonitrile, and the suspension was cooled to room temperature. The solid collected was then vacuum filtered to give compound I-15 (250 mg, 84%).

[0596] The following compounds were prepared in a similar manner from the appropriate lactam and alkylating reagent: I-14, I-16, I-17, I-19, I-20, I-21, I-22, I-37, I-47, I-5 7. I-58, I-60, I-62, I-65-67, I-70, I-94, I-96, I-98, I-100, I-112, I-114, I-116 and I-118..

[0597] Method F: Chiral separation of compound I-7 yielded compounds I-7-a and I-7-b.

[0598]

[0599] Compound I-7 (63 mg, 0.18 mmol) was dissolved in a mixture of methanol / ethanol and acetonitrile, and then purified by supercritical fluid chromatography using a Chiralpak AS-H, 20 x 250 mm, 5 μm column, eluting with 30% ethanol in CO2 at a flow rate of 50 mL / min and a pressure of 100 bar to give the title compound.

[0600] Peak 1-Chiral LC: retention time 3.21 min, Chiralpak AS, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min at 100 bar, and detected at 210 nm.

[0601] Peak 2-Chiral LC: retention time 7.34 min, Chiralpak AS, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min at 100 bar, and detected at 210 nm.

[0602] Method G: Chiral separation of compound I-8 yielded compounds I-8-a and I-8-b.

[0603]

[0604] Compound I-8 (96 mg, 0.26 mmol) was dissolved in a mixture of methanol, ethanol, DCM and acetonitrile, and then purified by supercritical fluid chromatography using a Chiralpak AS-H, 20 x 250 mm, 5 μm column, eluting with 30% ethanol in CO2 at a flow rate of 50 mL / min and a pressure of 100 bar to give the title compound.

[0605] Peak 1-Chiral LC: retention time 4.06 min, Chiralpak AS, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min at 100 bar, and detected at 210 nm.

[0606] Peak 2-Chiral LC: retention time 6.39 min, Chiralpak AS, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min at 100 bar, and detected at 210 nm.

[0607] Method H: Chiral separation of compound I-9 yielded compounds I-9-a and I-9-b.

[0608]

[0609] Compound I-9 (24 mg, 0.067 mmol) was dissolved in a mixture of ethanol and acetonitrile and then purified by supercritical fluid chromatography using a Chiralpak AS-H, 20 x 250 mm, 5 μm column, eluting with 30% ethanol in CO2 at a flow rate of 50 mL / min and a pressure of 100 bar to give the title compound.

[0610] Peak 1-Chiral LC: retention time 5.72 min, Chiralpak AS, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min at 100 bar, and detected at 210 nm.

[0611] Peak 2-Chiral LC: retention time 11.03 min, Chiralpak AS, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min at 100 bar, and detected at 210 nm.

[0612] Method I: Chiral separation of compound I-11 yielded compounds I-11-a and I-11-b.

[0613]

[0614] Compound I-11 (30 mg, 0.0844 mmol) was dissolved in a mixture of methanol, acetonitrile, IPA and formic acid, and then purified by supercritical fluid chromatography using a Chiralpak AD-H, 10 x 250 mm, 5 μm column, eluting with 40% methanol in CO2 at a flow rate of 15 mL / min to give the title compound.

[0615] Peak 1 - Chiral LC: retention time 11.38 min, Chiralpak AD-H, 4.6 x 250 mm, 5 μm, eluted with 40% methanol in CO2 at 4 mL / min at 100 bar, and detected at 210 nm.

[0616] Peak 2-Chiral LC: retention time 19.13 min, Chiralpak AD-H, 4.6 x 250 mm, 5 μm, eluted with 40% methanol in CO2 at 4 mL / min at 100 bar, and detected at 210 nm.

[0617] Method J: Chiral separation of compound I-20 yielded compounds I-20-a and I-20-b.

[0618]

[0619] Compound I-20 (124 mg, 0.322 mmol) was dissolved in a mixture of methanol (5 mL), ethanol (15 mL), and acetonitrile (10 mL), and then purified by supercritical fluid chromatography using a Chiralpak AS-H, 20 x 250 mm, 5 μm column, eluting with 30% ethanol in CO2 at a flow rate of 50 mL / min and a pressure of 100 bar to give the title compound.

[0620] Peak 1-Chiral LC: retention time 5.69 min, Chiralpak AS-H, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min and 100 bar, and detected at 210 nm.

[0621] Peak 2-Chiral LC: retention time 8.88 min, Chiralpak AS-H, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min and 100 bar, and detected at 210 nm.

[0622] Method K: Chiral separation of compound I-19 yielded compounds I-19-a and I-19-b.

[0623]

[0624] Compound I-19 (20 mg, 0.0545 mmol) was dissolved in a mixture of methanol, acetonitrile, and IPA, and then purified by flash column chromatography using a Chiralcel OJ-H, 20 x 250 mm, 5 μm column, eluting with a heptane solution of 15% ethanol at a flow rate of 18 mL / min to give the title compound.

[0625] Peak 1 - Chiral LC: retention time 34.09 min, Chiralcel OJ-H, 4.6 x 250 mm, 5 μm, eluted with 15% ethanol in heptane at 1 mL / min, and detected at 210 nm.

[0626] Peak 2-Chiral LC: retention time 43.34 min, Chiralcel OJ-H, 4.6 x 250 mm, 5 μm, eluted with 15% ethanol in heptane at 1 mL / min, and detected at 210 nm.

[0627] Method L: Chiral separation of compound I-21 yielded compounds I-21-a and I-21-b.

[0628]

[0629] Compound I-21 (112 mg, 0.305 mmol) was dissolved in a mixture of methanol (8 mL) and acetonitrile (2 mL), and then purified by supercritical fluid chromatography using a Chiralpak IB, 20 x 250 mm, 5 μm column, eluting with 20% ethanol in CO2 at a flow rate of 50 mL / min and a pressure of 100 bar to give the title compound.

[0630] Peak 1-Chiral LC: retention time 7.55 min, Chiralpak IB, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min and 100 bar, and detected at 210 nm.

[0631] Peak 2-Chiral LC: retention time 8.53 min, Chiralpak IB, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 2.4 mL / min and 100 bar, and detected at 210 nm.

[0632] Method M: Chiral separation of compound I-16 yielded compounds I-16-a and I-16-b.

[0633]

[0634] Compound I-16 (172 mg, 0.49 mmol) was dissolved in MeOH:DCM (1:1, 6 mg / mL) and purified by supercritical fluid chromatography using a Lux C3 column (20 mm x 250 mm x 5 μm). The elution was performed with 15% MeOH in CO2 at a flow rate of 50 mL / min and a pressure of 125 bar to give the title compound.

[0635] Peak 1 - Chiral LC: retention time 1.95 min, on Lux C3 (4.6 mm x 250 mm x 5 μm), eluted with 20% MeOH in CO2 with 0.2% NH3 modifier.

[0636] Peak 2-Chiral LC: retention time 2.24 min, on Lux C3 (4.6 mm x 250 mm x 5 μm), eluted with 20% MeOH in CO2 with 0.2% NH3 modifier.

[0637] Method N: Chiral separation of compound I-4 yielded compounds I-4-a and I-4-b.

[0638]

[0639] Compound I-4 (2.5 g, 7.0 mmol) was separated by SFC to obtain compounds I-4-a and I-4-b. Peak 1 (762 mg) and peak 2 (670 mg) were observed.

[0640] Chiral preparation conditions

[0641] Instrument: SFC-80 (Thar, Waters)

[0642] Column: AS 20*250mm, 10um (Daicel)

[0643] Columnarity: 40℃

[0644] Mobile phase: CO2 / MeOH (0.2% methanol ammonia) = 80 / 65

[0645] Flow rate: 80g / min

[0646] Back pressure: 100 bar

[0647] Detection wavelength: 220nm

[0648] Cycle time: 9.5 min

[0649] Sample solution: 2500 mg dissolved in 250 mL of methanol

[0650] Injection volume: 3mL

[0651] Method O: Chiral separation of compound I-5 yielded compounds I-5-a and I-5-b.

[0652] Compound I-5 (1.8 g) was separated by SFC to obtain compounds I-5-a and I-5-b. Peak 1 (486 mg) and peak 2 (290 mg) were observed.

[0653] Chiral preparation conditions

[0654] Instrument: SFC-80 (Thar, Waters)

[0655] Column: OJ 20*250mm, 10um (Daicel)

[0656] Column temperature: 35℃

[0657] Mobile phase: CO2 / MeOH (0.2% methanol ammonia) = 60 / 40

[0658] Flow rate: 80g / min

[0659] Back pressure: 100 bar

[0660] Detection wavelength: 214nm

[0661] Cycle time: 5.0 min

[0662] Sample solution: 1800 mg dissolved in 80 mL of methanol

[0663] Injection volume: 1.9 mL

[0664] Method P: Chiral separation of compound I-6 yielded compounds I-6-a and I-6-b.

[0665] Chiral preparation conditions

[0666] Instrument: SFC-80 (Thar, Waters)

[0667] Column: (R,R)Whelk-O1 20*250mm, 10um (Daicel)

[0668] Column temperature: 40℃

[0669] Mobile phase: CO2 / MEOH (1.0% methanol ammonia) = 60 / 40

[0670] Flow rate: 80g / min

[0671] Back pressure: 100 bar

[0672] Detection wavelength: 219nm

[0673] Cycle time: 7min

[0674] Sample solution: 75 mg dissolved in 10 mL of methanol

[0675] Injection volume: 1 mL

[0676] Method Q: Synthesis of intermediate Int-8

[0677]

[0678] DIPEA (2.4 g, 18.6 mmol) and propionic anhydride (5.9 g, 9.3 mmol) in 50% EtOAc solution (1.34 g, 7.4 mmol) were added to a solution of R-7 (1.34 g, 7.4 mmol) in DMA (10 mL) at 0 °C. The mixture was stirred for 10 min, and then R-6 (1.0 g, 6.2 mmol) was added. The mixture was stirred for 1 h, then quenched with an aqueous solution of NaHCO3, and the precipitate was filtered to give 5-bromo-N-(5-(pyridin-4-yl)-2H-1,2,4-triazol-3-yl)pentanamide (0.8 g, 40%).

[0679] NaH (60%, 136 mg, 3.4 mmol) was added to a solution of 5-bromo-N-(5-(pyridin-4-yl)-2H-1,2,4-triazol-3-yl)pentanamide (500 mg, 1.55 mmol) in DMF (5 mL) at 0 °C. The mixture was stirred for 30 min, then SEMCl (258 mg, 1.55 mmol) was added dropwise, and the mixture was stirred for another 30 min. The mixture was quenched with water and extracted with ethyl acetate (100 mL x 2). The combined extracts were concentrated under vacuum and purified by reversed-phase chromatography (eluting with 0–45% MeCN aqueous solution) to give Int-8 (220 mg, 38%).

[0680] Method R: Synthesis of compound I-23

[0681]

[0682] A solution of LDA in THF (2 mol / L, 6.0 mL, 12 mmol) was slowly added to a mixture of Int-8 (3.0 g, 8.0 mmol) in THF (30 mL) at -60 °C over 1.0 h, followed by the addition of R-8 (1.5 g, 8.0 mmol) in THF (30 mL). The reaction mixture was stirred at -60 °C for 2 h. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (100 mL x 2). The combined organic layers were dried, concentrated under vacuum, and the crude product (SiO2, DCM:MeOH = 20:1) was purified by rapid chromatography. The protected product (2.2 g, 57%) was collected, dissolved in DCM (15 mL), and treated with TFA (15 mL) at room temperature for 1 h. The reaction mixture was concentrated under vacuum to give the crude product, which was then treated with water (15 mL) and saturated NaHCO3 (80 mL). The solid precipitate was filtered, and the filter cake was concentrated under vacuum to obtain compound I-23 (1.5 g, 91%).

[0683] The following compounds were prepared in a similar manner using suitable intermediates prepared by method Q: I-25, I-27-34, I-36, I-41, I-48-53, I-55, I-56 and I-68.

[0684] Method S: Synthesis of compound I-24

[0685]

[0686] A solution of LDA in THF (2.0 M, 5.0 mL, 10 mmol) was added dropwise to a solution of Int-8 (2.5 g, 6.7 mmol) in dry THF (30 mL) at -78 °C over 30 min. The mixture was stirred at -78 °C for 1 h, and then a solution of R-9 (2.23 g, 10 mmol) in THF (5 mL) was added dropwise over 30 min. After stirring at -78 °C for 2 h, the reaction mixture was quenched with water and extracted with ethyl acetate (100 mL x 3). The combined extracts were washed with water and brine and concentrated under vacuum. The residue was purified by reversed-phase chromatography (eluting with 0–60% MeCN in NH4HCO3 solution) to give the alkylated product (2.3 g, 67%), which was dissolved in DCM (20 mL) and slowly treated with TFA (10 mL) at 0 °C. The mixture was then stirred at rt for 2 h, concentrated under vacuum, and then partitioned between a NaHCO3 aqueous solution (20 ml) and ethyl acetate (5 ml). The resulting precipitate was filtered, and the cake was washed with cold ethyl acetate to give compound I-24 (1.2 g, 70%).

[0687] Method T: Chiral separation of compound I-23 yielded compounds I-23-a and I-23-b.

[0688]

[0689] Compound I-23 (2.0 g) was dissolved in 140 mL of MeOH and separated by SFC using a Daicel OZ, 20 x 250 mm, 10 μm column. The title compound was obtained by elution with 45% MeOH (1% ammonia) in CO2 at a flow rate of 80 g / min, a pressure of 100 bar, and a temperature of 40 °C.

[0690] Peak 1–(650 mg) chiral LC: retention time 1.93 min, on Chiralcel OZ-H, 4.6 x 100 mm, 5 μm, eluted with 45% MeOH (0.2% ammonia) in CO2 at 4 mL / min.

[0691] Peak 2–(680 mg) chiral LC: retention time 2.5 min, eluted with 45% MeOH (0.2% ammonia) in CO2 at 4 mL / min on Chiralcel OZ-H, 4.6 x 100 mm, 5 μm.

[0692] Method U: Chiral separation of compound I-24 yielded compounds I-24-a and I-24-b.

[0693]

[0694] Compound I-24 (1.2 g) was dissolved in 160 mL of MeOH and separated by SFC using a Daicel AS, 20 x 250 mm, 10 μm column. The title compound was obtained by elution with 50% MeOH (1% ammonia) in CO2 at a flow rate of 80 g / min, a pressure of 100 bar, and a temperature of 40 °C.

[0695] Peak 1–(262 mg) chiral LC: retention time 1.54 min, eluted with 25% MeOH (0.2% ammonia) in CO2 at 4 mL / min on Chiralcel AS-H, 4.6 x 100 mm, 5 μm.

[0696] Peak 2–(255mg) chiral LC: retention time 2.1 min, eluted with 25% MeOH (0.2% ammonia) in CO2 at 4 mL / min on Chiralcel AS-H, 4.6 x 100 mm, 5 μm.

[0697] Method V: Synthesis of Compound I-59

[0698]

[0699] Int-1 (1.14 g, 1.55 mmol) was dissolved in CH₂Cl₂ (5 mL) at room temperature. Trifluoroacetic acid (2.0 mL, 26.1 mmol) was added, and the reaction mixture was stirred for 16 hours. The reaction mixture was concentrated under vacuum, and the crude product was dissolved in CH₂Cl₂ (20 mL), followed by the addition of saturated NaHCO₃ aqueous solution (10 mL) and water (10 mL). The organic layer was separated, and the aqueous layer was extracted with CH₂Cl₂ (2 x 20 mL). The organic layers were combined, dried (MgSO₄), filtered, and concentrated under vacuum to obtain the crude product. Formaldehyde (37%, 89 μL, 1.10 mmol) was added to the crude product (250 mg, 0.73 mmol) in anhydrous ethanol (3 mL), and the reaction mixture was stirred at 37 °C for 20 minutes. The crude product was purified by preparative HPLC to give the title compound (78 mg, 30%).

[0700] The following compounds, I-35 and I-64, were prepared in a similar manner.

[0701] Method W: Synthesis of compound I-76-a

[0702]

[0703] A solution of NaHMDS in THF (2M, 4.2mL, 8.44mmol) was added dropwise to a solution of Int-9 (3g, 7.67mmol) cooled to -78°C in 50mL dry THF under a N2 atmosphere, while maintaining the temperature below -65°C. The mixture was stirred at -78°C for 2 hours, and then a solution of 4-(bromomethyl)-1-chloro-2-fluorobenzene (4.26g, 19.18mmol) in 30mL dry THF was added. The mixture was stirred at -78°C until room temperature was reached over 12 hours, then cooled to -78°C and quenched with 80mL of saturated NaHCO3 aqueous solution. The aqueous phase was extracted with EtOAc (100mL x 5), and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by column chromatography to obtain the alkylated product (2.5 g, 4.69 mmol), which was dissolved in THF (30 mL) and treated with dropwise addition of HF-Pyr (2.32 g, 23.45 mmol). The mixture was stirred for 2 h, then treated with EtOAc (50 mL), concentrated under vacuum, and purified by column chromatography to obtain an alcohol (1.3 g, 3.1 mmol). The alcohol was dissolved in CH2Cl2 (15 mL), and Dess-Martin reagent (1.97 g, 4.65 mmol) and NaHCO3 (0.52 g, 6.21 mmol) were added. The mixture was stirred for 1 h, then treated with saturated NaS2O4 aqueous solution (30 mL) and saturated NaHCO3 aqueous solution (20 mL). The aqueous layer was extracted with EtOAc (5 x 50 mL), dried over Na2SO4, and concentrated under vacuum. The residue was purified by column chromatography to obtain Int-10 (1.16 g, 90%).

[0704] To a solution of Int-10 (260 mg, 0.62 mmol) in MeOH (5 mL), 3-(pyridazin-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-5-amine (183 mg, 0.62 mmol) and AcOH (0.5 mL) were added. After 2 h, NaBH3CN (79 mg, 1.25 mmol) was added, and the mixture was stirred at room temperature for 12 h. The mixture was quenched with ice water (10 mL) and extracted with EtOAc (5 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by column chromatography to give an amine (250 mg, 0.36 mmol), which was dissolved in THF / H2O (5 mL) and treated with LiOH (25 mg, 1.08 mmol) and H2O2 (75 mg, 2.17 mmol). The mixture was stirred at room temperature for 30 min, then quenched with a saturated aqueous solution of NH4Cl (20 mL) and extracted with EtOAc (5 x 40 mL). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum to give Int-11 (170 mg, 91%).

[0705] DIPEA (82 mg, 0.64 mmol) and propionic anhydride (203 mg, 0.64 mmol) were added to a solution of Int-11 (170 mg, 0.32 mmol) in DMA (4 mL). The mixture was stirred at room temperature for 1 h, treated with water (15 mL), and extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄, concentrated under vacuum, and the residue was purified by column chromatography to give a lactam (140 mg, 85%). The lactam (30 mg, 0.06 mmol) was dissolved in CH₂Cl₂ (2 mL) and treated with TFA (0.5 mL). The mixture was stirred at room temperature for 1 h and then concentrated under vacuum. The residue was purified by preparative-HPLC to give I-76-a (19 mg, 85%).

[0706] Chiral LC: retention time 3.73 min, eluted with 20% methanol (0.2% methanol ammonia) in CO2 at 3.00 mL / min at 140.1 bar on a Chiralpak Cellulose-SJ, 4.6*100 mm, 5 μm, and detected at 214 nm.

[0707] The following compounds were prepared in a similar manner: I-77-a, I-84-a, and I-85-a.

[0708] Method X: Chiral separation of compound I-75 yielded compounds I75-a and I-75-b.

[0709]

[0710] Compound I-75 was dissolved in CH2Cl2 and methanol and purified by supercritical fluid chromatography using a chiral ART SA, 21.2 x 250 mm, 5 μm column. The elution was performed with 40% methanol in CO2 at a flow rate of 50 mL / min and a pressure of 125 bar to give the title compound.

[0711] Peak 1 - Chiral LC: retention time 3.90 min, on chiral ART SA, 4.6 x 250 mm, 5 μm, eluted with 40% methanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 125 bar.

[0712] Peak 2 - Chiral LC: retention time 6.05 min, on chiral ART SA, 4.6 x 250 mm, 5 μm, eluted with 40% methanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 125 bar.

[0713] Method Y: Chiral separation of compound I-74 yielded compounds I74-a and I-74-b.

[0714]

[0715] Compound I-74 (300 mg, 0.79 mmol) was dissolved in methanol and then purified by supercritical fluid chromatography using a Chiralpak AD 20*250 mm, 10 μm (Daicel) column. The elution was performed with 0.2% methanol-ammonia in CO2 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0716] Peak 1 - Chiral LC: retention time 3.24 min, on a Chiralpak AD-H, 4.6*100 mm, 5 μm column, eluted with CO2 / MeOH (0.2% methanol ammonia) = 65 / 35, at 156 bar, 4 mL / min, and eluted at 214 nm.

[0717] Peak 2-Chiral LC: retention time 4.04 min, eluted with CO2 / MeOH (0.2% methanol ammonia) = 65 / 35 at 159 bar at 4 mL / min on a Chiralpak AD-H, 4.6*100 mm, 5 μm column, and detected at 214 nm.

[0718] Method Z: Chiral separation of compound I-82 yielded compounds I82-a and I-82-b.

[0719]

[0720] Compound I-82 (250 mg, 0.71 mmol) was dissolved in a mixture of methanol, acetonitrile, ethanol, and isopropylamine, and purified by supercritical fluid chromatography (SCLC) at Chiralpak AS-H, 20 x 250 mm, 5 μM. The compound was eluted with 20% ethanol in CO2 at a flow rate of 50 mL / min and a pressure of 99.5 bar to give the title compound. Peak 1 - Chiral LC: retention time 14.89 mins, Chiralpak AS, 4.6 x 250 mm, 5 μm, eluted with 20% ethanol in CO2 at 2.4 mL / min and 100 bar, and detected at 254 nm. Peak 2 - Chiral LC: retention time 15.14 mins, Chiralpak AS, 4.6 x 250 mm, 5 μm, eluted with 20% ethanol in CO2 at 2.4 mL / min and 100 bar, and detected at 254 nm.

[0721] Method AA: Chiral separation of compound I-54 yielded compounds I-54-a and I-54-b.

[0722]

[0723] Compound I-54 (90 mg, 0.236 mmol) was dissolved in methanol and then purified by supercritical fluid chromatography at OD 20*250 mm, 10 μm (Daicel), CO2 / MEOH (0.2% methanol ammonia) = 65 / 35, flow rate of 80 g / min and pressure of 100 bar to obtain the title compound.

[0724] Peak 1 - Chiral LC: retention time 2.33 min, at OD-H 4.6*100 mm 5 μm, CO2 / MEOH (0.2% methanol ammonia) = 65 / 35, 4 mL / min, at 120 bar, and detected at 214 nm.

[0725] Peak 1 - Chiral LC: retention time 2.62 min, at OD-H 4.6*100 mm 5 μm, CO2 / MEOH (0.2% methanol ammonia) = 65 / 35, 4 mL / min, at 120 bar, and detected at 214 nm.

[0726] Methods A and B: Chiral separation of compound I-46 yielded compounds I-46-a and I-46-b.

[0727]

[0728] Compound I-46 (100 mg, 0.272 mmol) was dissolved in methanol and then purified by supercritical fluid chromatography using a Chiralpak AS 20*250 mm, 10 μm column. The elution was performed with 50% methanol (1.0% methanol ammonia) in CO2 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0729] Peak 1 - Chiral LC: retention time 1.14 min, eluted with 50% methanol (1.0% methanol ammonia) in CO2 at 3 ml / min at 100 bar on a Chiralpak AS-3, 4.6*100 mm, 3 μm column, and detected at 214 nm.

[0730] Peak 2-Chiral LC: retention time 2.48 min, on a Chiralpak AS-3, 4.6*100 mm, 3 μm column, with 50% methanol (1.0% methanol ammonia) in CO2 at 3 ml / min at 100 bar, and detected at 214 nm. Method AC: Compound I-69 Chiral separation yielded compounds I-69-a and I-69-b.

[0731]

[0732] Compound I-69 (134 mg, 0.38 mmol) was dissolved in ethanol and purified using a Chiralpak AS-V, 76.5 x 300 mm, 20 μm microscope. The compound was then eluted with a heptane solution of 30% ethanol (with 0.5% isopropylamine as a modifier) ​​at a flow rate of 275 mL / min to give the title compound.

[0733] Peak 1-Chiral LC: retention time – 3.02 min, on Chirapak AS-H, 4.6 x 50 mm, 5 μm, eluted with 30% ethanol in CO2 (0.5% isopropylamine as modifier) ​​at 0.8 mL / min and detected at 254 nm.

[0734] Peak 2-Chiral LC: retention time – 4.44 min, on Chirapak AS-H, 4.6 x 50 mm, 5 μm, eluted with 30% ethanol in CO2 (0.5% isopropylamine as modifier) ​​at 0.8 mL / min and detected at 254 nm.

[0735] Method AD: Chiral separation of compound I-69 yielded compounds I-69-a and I-69-b.

[0736]

[0737] The compound (270 mg, 0.72 mmol) was dissolved in methanol (30 mL) and then purified by supercritical fluid chromatography. The compound was obtained by elution on a Chiralpak OJ 20*250 mm, 10 μm (Daicel) column with CO2 / EtOH (1.0% methanol ammonia) = 65 / 35 at a flow rate of 100 g / min and a pressure of 100 bar.

[0738] Peak 1 - Chiral LC: retention time 4.058 min, eluted with CO2 / EtOH [1% NH3 (7M MeOH solution)] = 80 / 20 on a Chiralpak OJ-3, 4.6*100 mm, 3 μm column at a flow rate of 3 mL / min at 2000 psi and detected at 214 nm.

[0739] Peak 2-Chiral LC: retention time 3.447 min, eluted with CO2 / EtOH [1% NH3 (7M MeOH solution)] = 80 / 20 at a flow rate of 3 mL / min at 2000 psi on a Chiralpak OJ-3, 4.6*100 mm, 3 μm column, and detected at 214 nm.

[0740] Method AE: Chiral separation of compound I-55 yielded compounds I-55-a and I-55-b.

[0741]

[0742] Compound I-55 (240 mg, 0.65 mmol) was dissolved in a mixture of methanol, acetonitrile, and formic acid, and then purified by supercritical fluid chromatography using Chiralpak AD-H, 10 x 250 mm, 5 μm, eluting with 30% isopropanol in CO2 at a flow rate of 15 mL / min to give the title compound.

[0743] Peak 1 - Chiral LC: retention time 7.05 min, Chiralpak AD-H, 4.6 x 250 mm, 5 μm, eluted with 35% isopropanol in CO2 at 4 mL / min.

[0744] Peak 2-Chiral LC: retention time 8.36 min, Chiralpak AD-H, 4.6 x 250 mm, 5 μm, eluted with 35% isopropanol in CO2 at 4 mL / min.

[0745] Method AF: Chiral separation of compound I-49 yielded compounds I-49-a and I-49-b.

[0746]

[0747] Compound I-49 (134 mg, 0.36 mmol) was dissolved in ethanol and purified by supercritical fluid chromatography using Lux A2, 21.2 mm x 250 mm, 5 μm. The elution was performed with 40% ethanol in CO2 (0.2% NH3 as a modifier) ​​at a pressure of 125 bar and a flow rate of 50 mL / min to give the title compound.

[0748] Peak 1-Chiral LC: retention time 1.43 min, Lux A2, 4.6 mm x 250 mm, 5 μm, eluted with 40% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 125 bar, and detected at 210-400 nm.

[0749] Peak 2-Chiral LC: retention time 1.69, on Lux A2, 4.6 mm x 250 mm, 5 μm, eluted with 40% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 125 bar, and detected at 210–400 nm.

[0750] Method AG: Chiral separation of compound I-48 yielded compounds I-48-a and I-48-b.

[0751]

[0752] Compound I-48 (100%, 165 mg, 0.449 mmol) was dissolved in ethanol and then purified by supercritical fluid chromatography using Lux A2, 21.2 mm x 250 mm, 5 μm. The elution was performed with 30% ethanol in CO2 (0.2% NH3 as a modifier) ​​at a pressure of 125 bar and a flow rate of 50 mL / min to give the title compound.

[0753] Peak 1 - Chiral LC: retention time 3.25, on Lux A2, 4.6 mm x 250 mm, 5 μm, eluted with 40% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min, and detected at 210-400 nm.

[0754] Peak 2-Chiral LC: retention time 3.78, on Lux A2, 4.6 mm x 250 mm, 5 μm, eluted with 40% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min, and detected in the range of 210–400 nm.

[0755] Method AH: Chiral separation of compound I-71 yielded compounds I-71-a and I-71-b.

[0756]

[0757] Compound I-71 (256 mg, 0.69 mmol) was dissolved in a mixture of CH2Cl2, ethanol, heptane, and isopropylamine, purified using a Chiralcel AS-V, 76.5 x 500 mm, 20 μm microscope, and eluted with a heptane solution of 15% ethanol (0.5% isopropylamine as a modifier) ​​at a flow rate of 275 mL / min to give the title compound.

[0758] Peak 1-Chiral LC: retention time 3.91 min, Chirapak AS-H, 4.6 x 50 mm, 5 μm, eluted with 15% ethanol in CO2 (0.5% isopropylamine as modifier) ​​at 2.4 mL / min and 100 bar, and detected at 254 nm.

[0759] Peak 2-Chiral LC: retention time 5.11 min, Chirapak AS-H, 4.6 x 50 mm, 5 μm, eluted with 15% ethanol in CO2 (0.5% isopropylamine as modifier) ​​at 2.4 mL / min and 100 bar, and detected at 254 nm.

[0760] Method AI: Chiral separation of compound I-41 yielded compounds I-41-a and I-41-b.

[0761]

[0762] Compound I-41 (181 mg, 0.46 mmol) was dissolved in ethanol and purified by chiral supercritical fluid chromatography using a Lux A2 column (21.2 mm x 250 mm, 5 μm). The compound was eluted with 45% ethanol in CO2 (0.2% NH3 as a modifier) ​​at a pressure of 100 bar and a flow rate of 50 mL / min to give the title compound.

[0763] Peak 1-Chiral LC: retention time 1.80 min, on Lux A2, 4.6 mm x 250 mm, 5 μm, eluted with 45% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min, and detected at 210-400 nm.

[0764] Peak 2-Chiral LC: retention time 2.14 min, Lux A2, 4.6 mm x 250 mm, 5 μm, eluted with 45% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min, and detected at 210-400 nm.

[0765] Method AJ: Chiral separation of compound I-72 yielded compounds I-72-a and I-72-b.

[0766]

[0767] Compound I-72 (152 mg, 0.41 mmol) was dissolved in methanol, acetonitrile, and formic acid, and then purified by supercritical fluid chromatography using Chiralpak AD-H, 10 x 250 mm, 5 μm, eluting with 40% methanol in CO2 at a flow rate of 15 mL / min to give the title compound.

[0768] Peak 1 - Chiral LC: retention time 8.47 min, Chiralpak AD-H, 4.6 x 250 mm, 5 μm, eluted with 40% methanol in CO2 at 4 mL / min.

[0769] Peak 2-Chiral LC: retention time 17.87 min, Chiralpak AD-H, 4.6 x 250 mm, 5 μm, eluted with 40% methanol in CO2 at 4 mL / min.

[0770] Method AK: Chiral separation of compound I-47 yielded compounds I-47-a and I-47-b.

[0771]

[0772] Compound I-47 (101 mg, 0.21 mmol) was dissolved in CH2Cl2 and methanol and purified by supercritical fluid chromatography using a Lux C3 column (21.2 x 250 mm, 5 μm). The compound was eluted with 30% ethanol in CO2 at a flow rate of 50 mL / min and a pressure of 100 bar to give the title compound.

[0773] Peak 1 - Chiral LC: retention time 2.20 min, Lux C3, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 4 mL / min and 125 bar.

[0774] Peak 2-Chiral LC: retention time 2.79 min, Lux C3, 4.6 x 250 mm, 5 μm, eluted with 30% ethanol in CO2 at 4 mL / min and 125 bar.

[0775] Method: Chiral separation and deprotection of AL:Int-12 yielded compounds I-66-a and I-66-b.

[0776]

[0777] Int-12 (220 mg, 0.43 mmol) was dissolved in acetonitrile and purified by HPLC using Chiralpak IG, 20 mm x 250 mm, 5 μm, eluting with an acetonitrile solution of 10% isopropanol containing 0.2% NH3 at a flow rate of 21 mL / min to give the title compound. Peak 1 was further purified by supercritical fluid chromatography using Lux A1, 21.2 x 250 mm, 5 μm, eluting with 50% methanol in CO2 (0.2% NH3 as a modifier) ​​at a flow rate of 50 mL / min at 125 bar to give the title compound.

[0778] Peak 1-Chiral LC: retention time 1.53 min, Amy-C, 4.6 mm x 250 mm, 5 μm, eluted with 60% methanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 125 bar, and detected at 254 nm.

[0779] Peak 2-Chiral LC: retention time 2.69 min, on Amy-C, 4.6 mm x 250 mm, 5 μm, eluted with 60% methanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 125 bar, and detected at 254 nm.

[0780] The isolated enantiomers (30 mg, 0.06 mmol) were dissolved in CH₂Cl₂ (2 mL) and treated with TFA (0.5 mL). The mixture was stirred at room temperature for 1 h and then concentrated under vacuum to give the title compound.

[0781] Method AM: Chiral separation of compound I-51 yielded compounds I-51-a and I-51-b.

[0782]

[0783] Compound I-51 (200 mg, 0.53 mmol) was dissolved in 30 mL of methanol and then purified by supercritical fluid chromatography using a Chiralpak AY 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / MeOH (0.2% methanol ammonia) = 40 / 60 at a flow rate of 100 g / min and a pressure of 100 bar to obtain the title compound.

[0784] Peak 1 - Chiral LC: retention time 2.87 min, eluted with 50% ethanol (1% methanol ammonia) in CO2 at 4.00 mL / min at 153.1 bar on a Chiralpak OD-H, 4.6*100 mm, 5 μm, and detected at 214 nm.

[0785] Peak 2-Chiral LC: retention time 3.64 min, eluted with 50% ethanol (1% methanol ammonia) in CO2 at 4.00 mL / min at 151.1 bar on a Chiralpak OD-H, 4.6*100 mm, 5 μm, and detected at 214 nm.

[0786] Method AN: Chiral separation of compound I-80 yielded compounds I-80-a and I-80-b.

[0787]

[0788] Compound I-80 (2.3 g, 6.5 mmol) was dissolved in methanol and then purified by supercritical fluid chromatography using a chiralpak OZ 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / ETOH (1.0% methanol ammonia) = 50 / 50 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0789] Peak 1 - Chiral LC: retention time 2.02 min, eluted with CO2 / ETOH (1.0% methanol ammonia) = 60 / 40 at a flow rate of 4 mL / min and 155 bar on a chiralpak OZ 4.6*100 mm, 5 μm column, and detected at 260 nm.

[0790] Peak 2-Chiral LC: retention time 2.61 min, eluted with CO2 / ETOH (1.0% methanol ammonia) = 60 / 40 at a flow rate of 4 mL / min and 154.7 bar on a chiralpak OZ 4.6*100 mm, 5 μm column, and detected at 260 nm.

[0791] Method: Chiral separation and deprotection of AO:Int-13 yielded compounds I-81-a and I-81-b.

[0792]

[0793] Int-13 (642 mg, 1.27 mmol) was dissolved in ethanol and CH2Cl2 (3:1) and then purified by supercritical fluid chromatography using a Chiralpak IG column (20 x 250 mm, 5 μm). The elution was performed with 30% ethanol in CO2 (0.2% NH3 as a modifier) ​​at a flow rate of 50 mL / min and a pressure of 100 bar to give the title compound.

[0794] Peak 1 - Chiral LC: retention time 2.6 min, eluted with 35% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 125 bar pressure on Chiralpak IG, 4.6 x 250 mm, 5 μm.

[0795] Peak 2-Chiral LC: retention time 3.02 min, eluted with 35% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 125 bar on Chiralpak IG, 4.6 x 250 mm, 5 μm.

[0796] The isolated enantiomers (30 mg, 0.06 mmol) were dissolved in CH₂Cl₂ (2 mL) and treated with TFA (0.5 mL). The mixture was stirred at room temperature for 1 h and then concentrated under vacuum to give the title compound.

[0797] Method AP: Chiral separation of compound I-37 yielded compounds I-37-a and I-37-b.

[0798]

[0799] Compound I-37 (285 mg, 0.75 mmol) was dissolved in methanol, acetonitrile, and formic acid, and purified by supercritical fluid chromatography at Chiralpak IC, 10 x 250 mm, 5 μm. The compound was eluted with 25% ethanol in CO2 at a flow rate of 15 mL / min and a pressure of 100 bar to give the title compound.

[0800] Peak 1 - Chiral LC: retention time 10.99 min, on Chiralpak IG, 4.6 x 250 mm, 5 μm, eluted with 25% ethanol in CO2 at 4 mL / min.

[0801] Peak 2-Chiral LC: retention time 13.35 min, on Chiralpak IG, 4.6 x 250 mm, 5 μm, eluted with 25% ethanol in CO2 at 4 mL / min.

[0802] Method AQ: Chiral separation of compound I-65 yielded compounds I-65-a and I-65-b.

[0803]

[0804] Compound I-65 (1.22 g, 3.28 mmol) was dissolved in methanol (90 mL) and then purified by supercritical fluid chromatography using a Chiralpak AS-H, 20*250 mm, 10 μm column. The elution was performed with CO2 / EtOH (1.0% methanol ammonia) = 65 / 35 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0805] Peak 1-Chiral LC: retention time 3.27 min, eluted with 20% ethanol in CO2 at a flow rate of 4 mL / min at 147.3 bar on a Chiralpak AS-H, 4.6 x 250 mm, 5 μm column, and detected at 260 nm.

[0806] Peak 2-Chiral LC: retention time 4.44 min, eluted with 20% ethanol at 149 bar with CO2 at a flow rate of 4 mL / min on a Chiralpak AS-H, 4.6 x 250 mm, 5 μm column, and detected at 260 nm. Method AR: Chiral analysis of compound I-67 Compounds I-67-a and I-67-b were obtained.

[0807]

[0808] Compound I-67 (1.50 g, 4.45 mmol) was dissolved in methanol (160 mL) and then purified by supercritical fluid chromatography using a Chiralpak AS-H, 20*250 mm, 10 μm column. The elution was performed with CO2 / EtOH (1.0% methanol ammonia) = 60 / 40 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0809] Peak 1-Chiral LC: retention time 1.86 min, eluted with 25% ethanol in CO2 at a flow rate of 4 mL / min at 149.4 bar on a Chiralpak AS-H, 4.6 x 100 mm, 5 μm column, and detected at 265 nm.

[0810] Peak 2-Chiral LC: retention time 2.32 min, eluted with 25% ethanol in CO2 at a flow rate of 4 mL / min at 149 bar on a Chiralpak AS-H, 4.6 x 100 mm, 5 μm column, and detected at 265 nm. Method AS: Chirality of compound I-39 Compounds I-39-a and I-39-b were isolated.

[0811]

[0812] Compound I-39 (690 mg, 2.04 mmol) was dissolved in methanol (45 mL) and purified by supercritical fluid chromatography using a Chiralpak AS-H, 20*250 mm, 10 μm column. The elution was performed with CO2 / ETOH (1.0% methanol ammonia) = 80 / 20 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0813] Peak 1 - Chiral LC: retention time 1.487 min, eluted with CO2 / MeOH [0.2% NH3 (7M MeOH solution)] = 80 / 20 at 3 mL / min at 2000 psi on a Chiralpak AS-H, 4.6*100 mm, 3 μm column, and detected at 214 nm.

[0814] Peak 2-Chiral LC: retention time 1.951 min, on a Chiralpak AS-H, 4.6*100 mm, 3 μm column, eluted with CO2 / MeOH [0.2% NH3 (7 M MeOH solution)] = 80 / 20 at 3 mL / min at 2000 psi, and detected at 214 nm.

[0815] Method AT: Chiral separation of compound I-60 yielded compounds I-60-a and I-60-b.

[0816]

[0817] Compound I-60 (40 mg, 0.1 mmol) was dissolved in methanol (3 mL) and then purified by supercritical fluid chromatography at 20*250 mm, 10 μm (Daicel) using CO2 / MEOH (1.0% methanol ammonia) = 60 / 40 at a flow rate of 80 g / min and a pressure of 100 bar to obtain the title compound.

[0818] Peak 1 - Chiral LC: retention time 1.71 min, eluted with CO2 / MeOH [0.2% NH3 (7M MeOH solution)] = 75 / 25 at a flow rate of 3 mL / min at 2000 psi on an AS 4.6 x 100 mm, 3 μm column, and detected at 214 nm.

[0819] Peak 2-Chiral LC: retention time 2.4 min, eluted with CO2 / MeOH [0.2% NH3 (7M MeOH solution)] = 75 / 25 at a flow rate of 4 mL / min at 152.9 bar on an AS 4.6 x 100 mm, 5 μm column, and detected at 214 nm.

[0820] Method AU: Chiral separation of compound I-38 yielded compounds I-38-a and I-38-b.

[0821]

[0822] Compound I-38 (115 mg, 0.30 mmol) was dissolved in a mixture of methanol and diethylamine and then purified by supercritical fluid chromatography using a Chiralcel OJ-H, 10 x 250 mm, 5 μm column. The elution was performed with 30% methanol in CO2 at a flow rate of 15 mL / min (0.2% diethylamine as a modifier) ​​to give the title compound.

[0823] Peak 1 - Chiral LC: retention time 3.09 min, eluted with 35% methanol in CO2 (0.2% diethylamine) at 4 mL / min on Chiralcel OJ-H, 4.6 x 250 mm, 5 μm, and detected at 220 nm.

[0824] Peak 2-Chiral LC: retention time 4.74 min, Chiralcel OJ-H, 4.6 x 250 mm, 5 μm, eluted with 35% methanol in CO2 (0.2% diethylamine) at 4 mL / min, and detected at 220 nm.

[0825] Method AV: Chiral separation of compound I-73 yielded compounds I-73-a and I-73-b.

[0826]

[0827] Compound I-73 (1.1 g, 3.13 mmol) was dissolved in a mixture of methanol and dichloromethane and then purified by supercritical fluid chromatography using a chiralpak OZ-H, 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / MEOH (0.2% methanol ammonia) = 45 / 55 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0828] Peak 1 - Chiral LC: retention time 3.93 min, eluted with 35% methanol in CO2 at 4.0 mL / min at 152.9 bar on Chiralpak OZ-H, 4.6 × 100 mm, 5 μm, and detected at 265 nm.

[0829] Peak 2-Chiral LC: retention time 5.49 min, eluted with 35% methanol in CO2 at 4.0 mL / min at 157.3 bar on Chiralpak OZ-H, 4.6*100 mm, 5 μm, and detected at 265 nm. Method AW: Chiral analysis of compound I-61 Compounds I-61-a and I-61-b were obtained.

[0830]

[0831] Compound I-61 (30 mg, 0.08 mmol) was dissolved in methanol (2 mL) and then purified by supercritical fluid chromatography using an AS 20*250 mm, 10 μm (Daicel) lens. The elution was performed with CO2 / MEOH (0.2% methanol ammonia) = 60 / 40 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0832] Peak 1 - Chiral LC: retention time 1.61 min, eluted with CO2 / MeOH (0.2% MeOH ammonia) = 75 / 25 at a flow rate of 4 mL / min at 153.2 bar on an AS 4.6 x 100 mm, 5 μm column, and detected at 214 nm.

[0833] Method AX: Chiral separation of compound I-40 yielded compounds I-40-a and I-40-b.

[0834]

[0835] Compound I-40 (25 mg, 0.07 mmol) was dissolved in methanol and then purified by supercritical fluid chromatography using a Chiralcel OD-H, 10 x 250 mm, 5 μm column. The compound was eluted with 15% methanol in CO2 at a flow rate of 15 mL / min and a pressure of 100 bar to give the title compound.

[0836] Peak 1-Chiral LC: retention time 8.13 min, eluted with 20% methanol in CO2 at 4 mL / min at 100 bar on a Chiralcel OD-H, 4.6 x 250 mm, 5 μm, and detected at 254 nm.

[0837] Peak 2-Chiral LC: retention time 10.60 min, Chiralcel OD-H, 4.6 x 250 mm, 5 μm, eluted with 20% methanol in CO2 at 4 mL / min at 100 bar, and detected at 254 nm.

[0838] Method AY: Chiral separation of compound I-34 yielded compounds I-34-a and I-34-b.

[0839]

[0840] Compound I-34 (1.70 g, 4.38 mmol) was dissolved in methanol (140 mL) and then purified by supercritical fluid chromatography using a Chiralpak OX, 20 x 250 mm, 10 μm column. The elution was performed with CO2 / MEOH (0.5% methanol ammonia) = 40 / 60 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0841] Peak 1 - Chiral LC: retention time 2.48 min, eluted with CO2 / MEOH (0.2% methanol ammonia) = 55 / 45 at a flow rate of 4 mL / min at 160.5 bar on a Chiralpak OX-H, 4.6 x 100 mm, 5 μm column, and detected at 265 nm.

[0842] Peak 2-Chiral LC: retention time 3.35 min, eluted with CO2 / MEOH (0.2% methanol ammonia) = 55 / 45 at a flow rate of 4 mL / min at 162.1 bar on a Chiralpak OX-H, 4.6 x 100 mm, 5 μm column, and detected at 265 nm.

[0843] Method AZ: Chiral separation and deprotection of compound Int-14 yielded compounds I-78-a and I-78-b.

[0844]

[0845] Compound Int-14 (1 g, 1.99 mmol) was dissolved in methanol (25 mL) and then purified by supercritical fluid chromatography using a Chiralpak OX, 20*250 mm, 10 μm column. The elution was performed with CO2 / MEOH (1.0% methanol ammonia) = 45 / 55 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0846] Peak 1-Chiral LC: retention time 4.2 min, eluted with 60% methanol in CO2 at a flow rate of 4 mL / min on a Chiralpak IG, 4.6 x 100 mm, 5 μm column, and detected at 260 nm.

[0847] Peak 2-Chiral LC: retention time 2.77 min, eluted with 60% methanol in CO2 at a flow rate of 4 mL / min on a Chiralpak IG, 4.6 x 100 mm, 5 μm column, and detected at 260 nm.

[0848] The isolated enantiomers were dissolved in CH2Cl2 and treated with TFA. The mixture was stirred at room temperature for 1 h, and then concentrated under vacuum to obtain the title compound.

[0849] Method BA: Chiral separation of compound I-79 yielded compounds I-79-a and I-79-b.

[0850]

[0851] Compound I-79 (700 mg, 1.9 mmol) was dissolved in a methanol mixture and then purified by supercritical fluid chromatography using a chiralpak AS 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / MEOH (0.4% methanol ammonia) = 50 / 50 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0852] Peak 1 - Chiral LC: retention time 2.17 min, eluted with CO2 / MEOH (0.2% methanol ammonia) = 65 / 35 at a flow rate of 4 mL / min and 145.5 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 260 nm.

[0853] Peak 2-Chiral LC: retention time 3.61 min, eluted with CO2 / MEOH (0.2% methanol ammonia) = 65 / 35 at a flow rate of 4 mL / min and 150.5 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 260 nm.

[0854] Method BB: Chiral separation of compound I-32 yielded compounds I-32-a and I-32-b.

[0855]

[0856] Compound I-32 (1500 mg, 4.04 mmol) was dissolved in methanol (30 mL) and then purified by supercritical fluid chromatography using an OX, 20*250 mm, 10 μm column. The elution was performed with CO2 / MEOH (0.2% methanol ammonia) = 45 / 55 at a flow rate of 80 g / min and a pressure of 100 bar to obtain the title compound.

[0857] Peak 1 - Chiral LC: retention time 2.22 min, eluted with 0.2% methanol in CO2 at a flow rate of 4 mL / min at 157.3 bar on a Chiralpak OX-H, 4.6 x 100 mm, 5 μm column, and detected at 260 nm.

[0858] Peak 2-Chiral LC: retention time 2.99 min, eluted with 0.2% methanol in CO2 at a flow rate of 4 mL / min at 158.1 bar on a Chiralpak OX-H, 4.6 x 250 mm, 5 μm column, and detected at 260 nm.

[0859] Method BC: Chiral separation of compound I-36 yielded compounds I-36-a and I-36-b.

[0860]

[0861] Compound I-36 (580 mg, 1.56 mmol) was dissolved in a mixture of 80 mL of methanol and dichloromethane, and then purified by supercritical fluid chromatography using a Chiralpak OX 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / (MeOH / ACN (0.2% methanol ammonia) = 1:1) = 55 / 45 at a flow rate of 110 g / min and a pressure of 100 bar to give the title compound.

[0862] Peak 1 - Chiral LC: retention time 2.29 min, eluted with 45% methanol (0.2% methanol ammonia) in CO2 at 4.00 mL / min at 163.8 bar on a Chiralpak OX-H, 4.6 × 100 mm, 5 μm, and detected at 214 nm.

[0863] Peak 2-Chiral LC: retention time 3.02 min, eluted with 45% methanol (0.2% methanol ammonia) at 164.7 bar with CO2 at 4.00 mL / min on a Chiralpak OX-H, 4.6 × 100 mm, 5 μm, and detected at 214 nm.

[0864] Method BD: Chiral separation of compound I-89 yielded compounds I-89-a and I-89-b.

[0865]

[0866] Compound I-89 (800 mg, 2.1 mmol) was dissolved in methanol and then eluted on a Chiralpak AS 20*250 mm, 10 μm (Daicel) column with CO2 / MeOH:acetonitrile (3:2) / (0.2% methanol ammonia) = 50 / 50 at a flow rate of 80 g / min and a pressure of 100 bar to obtain the title compound.

[0867] Peak 1 - Chiral LC: retention time 1.98 min, eluted with CO2 / MeOH:acetonitrile:MeOHammonia (3:2:0.2%) = 65 / 35 at a flow rate of 4 mL / min and 146.5 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 255 nm.

[0868] Peak 2-Chiral LC: retention time 3.26 min, eluted with CO2 / MeOH:acetonitrile:MeOHammonia (3:2:0.2%) = 65 / 35 at a flow rate of 4 mL / min and 145.5 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 255 nm.

[0869] Method BE: Chiral separation of compound I-33 yielded compounds I-33-a and I-33-b.

[0870]

[0871] Compound I-33 (800 mg, 2.25 mmol) was dissolved in methanol (25 mL) and then purified by supercritical fluid chromatography using an OX, 20*250 mm, 10 μm column. The elution was performed with CO2 / MEOH (0.2% methanol ammonia) = 50 / 50 at a flow rate of 80 g / min and a pressure of 100 bar to obtain the title compound.

[0872] Peak 1 - Chiral LC: retention time 1.64 min, eluted with CO2 / MEOH (0.2% methanol ammonia) = 55 / 45 at a flow rate of 4 mL / min at 163.2 bar on a Chiralpak OX-H, 4.6 x 100 mm, 5 μm column, and detected at 260 nm.

[0873] Peak 2-Chiral LC: retention time 2.17 min, eluted with CO2 / MEOH (0.2% methanol ammonia) = 55 / 45 at a flow rate of 4 mL / min at 156.6 bar on a Chiralpak OX-H, 4.6 x 250 mm, 5 μm column, and detected at 260 nm.

[0874] Method BF: Chiral separation of compound I-31 yielded compounds I-31-a and I-31-b.

[0875]

[0876] Compound I-31 (480 mg, 1.25 mmol) was dissolved in a mixture of 25 mL of methanol and dichloromethane and then purified by supercritical fluid chromatography using a Chiralpak OX 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / (MeOH (0.2% methanol ammonia) = 50 / 50) at a flow rate of 100 g / min and a pressure of 100 bar to give the title compound.

[0877] Peak 1 - Chiral LC: retention time 1.92 min, eluted with 45% methanol (0.2% methanol ammonia) in CO2 at 3.00 mL / min at 154.4 bar on a Chiralpak OX-H, 4.6*100 mm, 5 μm, and detected at 214 nm.

[0878] Peak 2-Chiral LC: retention time 2.64 min, eluted with 45% methanol (0.2% methanol ammonia) in CO2 at 3.00 mL / min at 148.7 bar on a Chiralpak OX-H, 4.6*100 mm, 5 μm, and detected at 214 nm.

[0879] Method BG: Chiral separation of compound I-29 yielded compounds I-29-a and I-29-b.

[0880]

[0881] Compound I-29 (1900 mg, 4.74 mmol) was dissolved in methanol and dichloromethane and then purified by supercritical fluid chromatography using a Chiralpak OZ 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / (MeOH / ACN (0.2% methanol ammonia) = 1:1) = 45 / 55, at a flow rate of 120 g / min and a pressure of 100 bar to give the title compound.

[0882] Peak 1 - Chiral LC: retention time 2.76 min, eluted with 55% methanol (0.2% methanol ammonia) in CO2 at 3.0 mL / min at 155.6 bar on Chiralpak OZ-H, 4.6*100 mm, 5 μm, and detected at 214 nm.

[0883] Peak 2-Chiral LC: retention time 4.53 min, eluted with 55% methanol (0.2% methanol ammonia) in CO2 at 3.0 mL / min at 152.9 bar on Chiralpak OZ-H, 4.6*100 mm, 5 μm, and detected at 214 nm.

[0884] Method BH: Chiral separation of compound I-28 yielded compounds I-28-a and I-28-b.

[0885]

[0886] Compound I-28 (1300 mg, 3.38 mmol) was dissolved in methanol and then purified by supercritical fluid chromatography using a Chiralpak OX 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / MeOH (0.2% methanol ammonia) = 45 / 55 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0887] Peak 1 - Chiral LC: retention time 1.92 min, eluted with 45% methanol (0.2% methanol ammonia) in CO2 at 3.00 mL / min at 154.4 bar on a Chiralpak OX-H, 4.6 × 100 mm, 5 μm, and detected at 265 nm.

[0888] Peak 2-Chiral LC: retention time 2.64 min, eluted with 45% methanol (0.2% methanol ammonia) in CO2 at 3.00 mL / min at 148.7 bar on a Chiralpak OX-H, 4.6*100 mm, 5 μm, and detected at 265 nm.

[0889] Method BI: Chiral separation of compound I-90 yielded compounds I-90-a and I-90-b.

[0890]

[0891] Compound I-90 (940 mg, 2.6 mmol) was dissolved in methanol and then purified by supercritical fluid chromatography using a chiralpak AS 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / MEOH (0.2% methanol ammonia) = 50 / 50 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0892] Peak 1 - Chiral LC: retention time 1.36 min, eluted with CO2 / MEOH (0.2% methanol ammonia) = 60 / 40 at a flow rate of 4 mL / min and 147.8 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 260 nm.

[0893] Peak 2-Chiral LC: retention time 1.89 min, eluted with CO2 / MEOH (0.2% methanol ammonia) = 60 / 40 at a flow rate of 4 mL / min and 147.8 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 260 nm.

[0894] Method BJ: Chiral separation of compound I-91 yielded compounds I-91-a and I-91-b.

[0895]

[0896] Compound I-91 (1.2 g, 3.1 mmol) was dissolved in methanol and then purified by supercritical fluid chromatography using a chiralpak AS 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / ETOH (1.0% methanol ammonia) = 50 / 50 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0897] Peak 1 - Chiral LC: retention time 3.05 min, eluted with CO2 / ETOH (1.0% methanol ammonia) = 65 / 35 at a flow rate of 3 mL / min and 138.9 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 265 nm.

[0898] Peak 2-Chiral LC: retention time 3.99 min, eluted with CO2 / ETOH (1.0% methanol ammonia) = 65 / 35 at a flow rate of 3 mL / min and 140.6 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 265 nm.

[0899] Method BK: Chiral separation of compound Int-15, followed by deprotection, yielded compounds I-93-a and I-93-b.

[0900]

[0901] Int-15 (220 mg, 0.34 mmol) was dissolved in methanol and isopropanol and purified by supercritical fluid chromatography using a Chiralpak AD-H, 10 x 250 mm, 5 μm column, eluting with 30% isopropanol in CO2 at a flow rate of 15 mL / min to give the title compound.

[0902] Peak 1 - Chiral LC: retention time 1.78, Chiralpak AD-H, 10 x 250 mm, 5 μm, eluted with 30% isopropanol in CO2 at 4 mL / min, and detected at 254 nm.

[0903] Peak 2-Chiral LC: retention time 4.10, Chiralpak AD-H, 10 x 250 mm, 5 μm, eluted with 30% isopropanol in CO2 at 4 mL / min, detected at 254 nm.

[0904] The isolated enantiomers were dissolved in CH2Cl2, treated with TFA, and then concentrated under vacuum to obtain the title compound.

[0905] Method BL: Chiral separation of compound Int-16, followed by deprotection, yielded compounds I-92-a and I-92-b.

[0906]

[0907] Int-16 (290 mg, 0.54 mmol) was dissolved in ethanol and purified by supercritical fluid chromatography using a Lux A1 column (21.2 mm x 250 mm, 5 μm). The elution was performed with 50% ethanol in CO2 at a flow rate of 50 mL / min and a pressure of 100 bar to give the title compound.

[0908] Peak 1-Chiral LC: retention time 1.20, on Amy-C, 4.6 mm x 250 mm, 5 μm, eluted with 50% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 100 bar, and detected at 210–400 nm.

[0909] Peak 2-Chiral LC: retention time 5.65, on Amy-C, 4.6 mm x 250 mm, 5 μm, eluted with 50% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min and 100 bar, and detected at 210–400 nm.

[0910] The isolated enantiomers were dissolved in CH2Cl2, treated with TFA, and then concentrated under vacuum to obtain the title compound.

[0911] Method BM: Chiral separation of compound I-27 yielded compounds I-27-a and I-27-b.

[0912]

[0913] Compound I-27 (1060 mg, 2.87 mmol) was dissolved in methanol and then purified by supercritical fluid chromatography using a Chiralpak OZ 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / (MeOH (0.2% methanol ammonia) = 45 / 55) at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0914] Peak 1 - Chiral LC: retention time 2.463 min, Chiralpak OZ-3, 4.6*100 mm, 3 μm, eluted with 35% methanol (0.2% NH3 (7 M methanol solution)) in CO2 at 3.0 mL / min at 137.9 bar, and detected at 214 nm.

[0915] Peak 2-Chiral LC: retention time 3.877 min, Chiralpak OZ-3, 4.6*100 mm, 3 μm, eluted with 35% methanol (0.2% NH3 (7 M methanol solution)) in CO2 at 3.0 mL / min at 137.9 bar, and detected at 214 nm.

[0916] Method BN: Chiral separation of compound I-88 yielded compounds I-88-a and I-88-b.

[0917]

[0918] Compound I-88 (1.1 g, 2.8 mmol) was dissolved in a mixture of methanol and dichloromethane and then purified by supercritical fluid chromatography using a chiralpak AS 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / ETOH (0.5% methanol ammonia) = 55 / 45 at a flow rate of 80 g / min and a pressure of 100 bar to give the title compound.

[0919] Peak 1 - Chiral LC: retention time 1.49 min, eluted with CO2 / EtOH (1% methanol ammonia) = 60 / 40 at a flow rate of 3 mL / min and 147.8 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 214 nm.

[0920] Peak 2-Chiral LC: retention time 1.86 min, eluted with CO2 / EtOH (1% methanol ammonia) = 60 / 40 at a flow rate of 3 mL / min and 149.5 bar on a Chiralpak AS 4.6*100 mm, 5 μm column, and detected at 214 nm.

[0921] Method BO: Chiral separation of compound I-87 yielded compounds I-87-a and I-87-b.

[0922]

[0923] Compound I-87 (1.5 g, 4.0 mmol) was dissolved in a mixture of methanol and dichloromethane and then purified by supercritical fluid chromatography using a chiralpak AS-H, 20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / MEOH (0.2% methanol ammonia) = 70 / 30 at a flow rate of 100 g / min and a pressure of 100 bar to give the title compound.

[0924] Peak 1 - Chiral LC: retention time 2.47 min, Chiralpak AS-H, 4.6*100 mm, 5 μm, eluted with 25% methanol in CO2 at 3.0 mL / min at 140.1 bar, and detected at 214 nm.

[0925] Peak 2-Chiral LC: retention time 2.87 min, eluted with 25% methanol in CO2 at 3.0 mL / min at 140.4 bar on Chiralpak AS-H, 4.6 × 100 mm, 5 μm, and detected at 214 nm. Method BP: Chirality of compound Int-17 Separation, followed by deprotection, yielded compounds I-26-a and I-26-b.

[0926]

[0927] Int-17 (3000 mg, 6.1 mmol) was dissolved in methanol and purified by supercritical fluid chromatography using a chiralpak RR WHELK 20*250 mm, 10 μm column. The elution was performed with 25% ethanol in CO2 at a flow rate of 80 g / min and a pressure of 150.4 bar to give the title compound.

[0928] Peak 1 - Chiral LC: retention time 2.73 min, eluted with 25.0% ethanol in CO2 at 3 mL / min at 100 bar on a Chiralpak RR-Whelk-O1, 4.6 x 250 mm, 5 μm, and detected at 214 nm.

[0929] Peak 2-Chiral LC: retention time 3.29 min, Chiralpak RR-Whelk-O1, 4.6 x 250 mm, 5 μm, eluted with 25.0% ethanol in CO2 at 3 mL / min at 100 bar, and detected at 214 nm.

[0930] The isolated enantiomers were dissolved in CH2Cl2, treated with TFA, and then concentrated under vacuum to obtain the title compound.

[0931] Method BQ: Chiral separation of compound I-25 yielded compounds I-25-a and I-25-b.

[0932]

[0933] Compound I-25 (930 mg, 2.45 mmol) was dissolved in methanol and dichloromethane and purified by supercritical fluid chromatography using a chiralpak IC-320 x 250 mm, 10 μm column. The elution was performed with 40% ethanol in CO2 at a flow rate of 100 g / min and a pressure of 150.4 bar to give the title compound.

[0934] Peak 1 - Chiral LC: retention time 2.783 min, eluted with 40.0% ethanol in CO2 at 3 mL / min at 100 bar on a Chiralpak IC-3 4.5*100 mm, 5 μm, and detected at 214 nm.

[0935] Peak 2-Chiral LC: retention time 3.382 min, eluted with 40.0% ethanol in CO2 at 3 mL / min at 100 bar on a Chiralpak IC-3 4.6*100 mm, 5 μm, and detected at 214 nm.

[0936] Method BR: Chiral separation of compound Int-18, followed by deprotection, yielded compounds I-86-a and I-86-b.

[0937]

[0938] Compound Int-18 (2 g, 3.82 mmol) was dissolved in MeOH and then purified by supercritical fluid chromatography using a chiralpak OX-H column (4.6 × 100 mm, 5 μm column). The compound was eluted with 35% methanol in CO2 at a flow rate of 3 mL / min and a pressure of 149.7 bar to give the title compound.

[0939] Peak 1-Chiral LC: retention time 1.91 min, Chiralpak OX, 4.6*100 mm, 5 μm, eluted with 35% methanol in CO2 at 3 mL / min and 149.7 bar, and detected at 230 nm.

[0940] Peak 2-Chiral LC: retention time 2.26 min, Chiralpak OX, 4.6*100 mm, 5 μm, eluted with 35% methanol in CO2 at 3 mL / min and 145.2 bar, and detected at 230 nm.

[0941] The isolated enantiomers were dissolved in CH2Cl2, treated with TFA, and then concentrated under vacuum to obtain the title compound.

[0942] Method BS: Chiral separation of compound I-104 yielded compounds I-104-a and I-104-b.

[0943]

[0944] Compound I-104 (370 mg, 1.0 mmol) was dissolved in a methanol mixture and then purified by supercritical fluid chromatography using a chiralpak OZ20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / (MeOH / MeCN (0.2% methanol ammonia) = 9:1) = 50 / 50 at a flow rate of 120 g / min and a pressure of 100 bar to give the title compound.

[0945] Peak 1 - Chiral LC: retention time 2.695 min, Chiralpak OZ, 4.6 × 100 mm, 5 μm, eluted with 45% methanol (0.2% methanol ammonia) in CO2 at 3.0 mL / min at 2000 psi, and detected at 214 nm.

[0946] Peak 2-Chiral LC: retention time 3.574 min, Chiralpak OZ, 4.6*100 mm, 5 μm, eluted with 45% methanol (0.2% methanol ammonia) in CO2 at 3.0 mL / min at 2000 psi, and detected at 214 nm.

[0947] Method BT: Chiral separation of compound I-105 yielded compounds I-105-a and I-105-b.

[0948]

[0949] Compound I-105 (330 mg, 0.92 mmol) was dissolved in a mixture of methanol and dichloromethane (80 mL) and then purified by supercritical fluid chromatography using a chiralpak AS-H 20*250 mm, 10 μm (Regis) column. The elution was performed with CO2 / IPA (0.5% methanol-ammonia) = 40 / 60 at a flow rate of 100 g / min and a pressure of 100 bar to give the title compound.

[0950] Peak 1 - Chiral LC: retention time 2.062 min, on a Chiralpak AS-3 4.6*100 mm, 3 μm, eluted with 35% IPA [1% NH3 (7 M MeOH solution)] in CO2 at 3 mL / min at 2000 psi, and detected at 214 nm.

[0951] Peak 2-Chiral LC: retention time 2.781 min, on a Chiralpak AS-H 4.6*100 mm, 3 μm, eluted with 35% IPA [1% NH3 (7M MeOH solution)] in CO2 at 3.0 mL / min at 2000 psi, and detected at 214 nm.

[0952] Method BU: Chiral separation of compound I-106 yielded compounds I-106-a and I-106-b.

[0953]

[0954] Compound I-106 (450 mg, 1.2 mmol) was dissolved in methanol and dichloromethane (55 mL), and then purified by supercritical fluid chromatography using an OZ20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / (MeOH / CAN (0.2% methanol ammonia) = 1:1) = 50 / 50 at a flow rate of 120 g / min and a pressure of 100 bar to give the title compound.

[0955] Peak 1 - Chiral LC: retention time 1.984 min, eluted with 45% MeOH / MeCN = 3 / 2 [0.2% NH3 (7M MeOH solution)] in CO2 at a flow rate of 3 mL / min and 2000 psi on a chiralpak OZ 4.6*100 mm, 5 μm column, and detected at 214 nm.

[0956] Peak 2-Chiral LC: retention time 2.682 min, eluted with 45% MeOH / MeCN = 3 / 2 [0.2% NH3 (7M MeOH solution)] in CO2 at a flow rate of 3 mL / min and 2000 psi on a chiralpak OZ 4.6*100 mm, 5 μm column, and detected at 214 nm.

[0957] Method BV: Chiral separation of compound I-107 yielded compounds I-107-a and I-107-b.

[0958]

[0959] Compound I-107 (550 mg, 1.6 mmol) was dissolved in methanol and dichloromethane (80 mL) and purified by supercritical fluid chromatography using a chiralpak AS 20*250 mm, 10 μm (Daicel) spectrometer. The elution was performed with CO2 / MeOH (0.2% methanol-ammonia) = 60 / 40 at a flow rate of 100 g / min and a pressure of 100 bar to give the title compound.

[0960] Peak 1 - Chiral LC: retention time 2.404 min, on a Chiralpak AS-3 4.6*100 mm, 3 μm, eluted with 20% IPA [1% NH3 (7 M MeOH solution)] in CO2 at 3 mL / min at 2000 psi, and detected at 214 nm.

[0961] Peak 2-Chiral LC: retention time 2.891 min, on a Chiralpak AS-3 4.6*100 mm, 3 μm, eluted with 20.0% IPA [1% NH3 (7 M MeOH solution)] in CO2 at 3 mL / min at 2000 psi, and detected at 214 nm.

[0962] Method BW: Chiral separation of compound I-113 yielded compounds I-113-a and I-113-b.

[0963]

[0964] Compound I-113 (800 mg, 2.21 mmol) was dissolved in methanol (60 mL) and purified by supercritical fluid chromatography using an AS 20*250 mm, 10 μm (Regis) lens. The elution was performed with CO2 / MeOH (0.2% methanol ammonia) = 60 / 40 at a flow rate of 100 g / min and a pressure of 100 bar to give the title compound.

[0965] Peak 1 - Chiral LC: retention time 1.949 min, eluted with 20% IPA [0.2% NH3 (7M MeOH solution)] in CO2 at 3 mL / min at 2000 psi on a Chiralpak OJ-3 4.6*100 mm, 3 μm, and detected at 214 nm.

[0966] Peak 2-Chiral LC: retention time 2.369 min, eluted with 20% IPA [0.2% NH3 (7M MeOH solution)] in CO2 at 3 mL / min at 2000 psi on a Chiralpak OJ-3 4.6*100 mm, 3 μm, and detected at 214 nm.

[0967] Method BV: Chiral separation of compound I-119 yielded compounds I-119-a and I-119-b.

[0968]

[0969] Compound I-119 (450 mg, 1.2 mmol) was dissolved in methanol and dichloromethane (55 mL), and then purified by supercritical fluid chromatography using an OZ20*250 mm, 10 μm (Daicel) column. The elution was performed with CO2 / (MeOH / CAN (0.2% methanol ammonia) = 1:1) = 50 / 50 at a flow rate of 120 g / min and a pressure of 100 bar to give the title compound.

[0970] Peak 1 - Chiral LC: retention time 1.984 min, eluted with 45% MeOH / MeCN = 3 / 2 [0.2% NH3 (7M MeOH solution)] in CO2 at a flow rate of 3 mL / min and 2000 psi on a chiralpak OZ 4.6*100 mm, 5 μm column, and detected at 214 nm.

[0971] Peak 1 - Chiral LC: retention time 2.682 min, eluted with 45% MeOH / MeCN = 3 / 2 [0.2% NH3 (7M MeOH solution)] in CO2 at a flow rate of 3 mL / min and 2000 psi on a chiralpak OZ 4.6*100 mm, 5 μm column, and detected at 214 nm.

[0972] Method BX: Chiral separation of compound Int-19, followed by deprotection, yielded compounds I-83-a and I-83-b.

[0973]

[0974] Int-19 (460 mg, 0.70 mmol) was dissolved in ethanol and purified by supercritical fluid chromatography using a Lux A1, 21.2 mm x 250 mm, 5 μm lens. The compound was eluted with 50% ethanol in CO2 at a flow rate of 50 mL / min and a pressure of 125 bar to give the title compound.

[0975] Peak 1: Chiral LC: retention time 1.10 min, on Amy-C, 4.6 mm x 250 mm, 5 μm, eluted with 50% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min, and detected at 210-400 nm.

[0976] Peak 2: Chiral LC: retention time 2.84 min, on Amy-C, 4.6 mm x 250 mm, 5 μm, eluted with 50% ethanol in CO2 (0.2% NH3 as modifier) ​​at 4 mL / min, and detected at 210-400 nm.

[0977] The isolated enantiomers were dissolved in CH2Cl2, treated with TFA, and then concentrated under vacuum to obtain the title compound.

[0978] Example 2: Characterization of the compound.

[0979] LCMS method:

[0980] Analytical LC / MS analysis method A:

[0981] ESI + / - ion mode 150-850 Da

[0982] Column: Phenomenex Kinetix-XB C18, Part No.00D-4498-AN, 2.1x 100mm, 1.7μm

[0983] Temperature: 40℃

[0984] gradient:

[0985] 0 95％ 5％ 0.6 5.30 0％ 100％ 0.6 5.80 0％ 100％ 0.6 5.82 95％ 5％ 0.6 7.00 95％ 5％ 0.6

[0986] Analytical LC / MS analysis method B:

[0987] ESI + / - ion mode 150-850 Da

[0988] Column: Phenomenex Gemini-NX C18, Part No. 00D-4453-B0, 2.0 x 100 mm, 3.0 μm

[0989] Temperature: 40℃

[0990] gradient:

[0991] 0 95％ 5％ 0.6 5.50 0％ 100％ 0.6 5.90 0％ 100％ 0.6 5.92 95％ 5％ 0.6 7.00 95％ 5％ 0.6

[0992] Analytical LC / MS analysis method C:

[0993] ESI + / - ion mode 100-1000 Da

[0994] Column: Waters BEH TM C18,Part No.186002352,2.1x 100mm,1.7μm

[0995] Temperature: 40℃

[0996] gradient:

[0997] 0 95％ 5％ 0.6 5.30 0％ 100％ 0.6 5.80 0％ 100％ 0.6 5.82 95％ 5％ 0.6 7.00 95％ 5％ 0.6

[0998] Analytical LC / MS analysis method D:

[0999] ESI + / - ion mode 100-1000 Da

[1000] Column: XBridge C18, 3.5μm 4.6x50mm

[1001] Temperature: 40℃

[1002] gradient:

[1003] 0.00 95％ 5％ 2.0 1.20 5％ 95％ 2.0 3.00 5％ 95％ 2.0

[1004] Analytical LC / MS analysis method E:

[1005] ESI + / - ion mode 100-1000 Da

[1006] Column: XBridge SB-C18, 3.5μm, 4.6x50mm

[1007] Temperature: 40℃

[1008] gradient:

[1009] 0.00 95％ 5％ 2.0 1.40 5％ 95％ 2.0 4.30 5％ 95％ 2.0

[1010] Analytical LC / MS analysis method F:

[1011] ESI + / - ion mode 100-1000 Da

[1012] Column: Sunfire C18, 3.5μm 4.6x50mm

[1013] Temperature: 50℃

[1014] gradient:

[1015] 0.00 95％ 5％ 2.0 1.40 5％ 95％ 2.0 3.00 5％ 95％ 2.0

[1016] Analytical LC / MS analysis method G:

[1017] ESI + / - ion mode 100-1000 Da

[1018] Column: Waters BEH TM C18,Part No.186005297,1.7μm 2.1x50mm

[1019] Temperature: 40℃

[1020] gradient:

[1021]

[1022] Analytical LC / MS method H:

[1023] ESI + / - ion mode 100-1000 Da

[1024] Column: XBridge C18, 3.5μm 4.6x50mm

[1025] Temperature: 45℃

[1026] gradient:

[1027] 0.00 95％ 5％ 2.0 1.30 5％ 95％ 2.0 3.00 5％ 95％ 2.0

[1028] Analytical LC / MS Method I:

[1029] ESI + / - ion mode 100-1000 Da

[1030] Column: XBridge C18, 3.5μm 4.6x50mm

[1031] Temperature: 50℃

[1032] gradient:

[1033] 0.00 95％ 5％ 2.0 1.40 5％ 95％ 2.0 3.00 5％ 95％ 2.0

[1034] Analytical LC / MS method J:

[1035] ESI + / - ion mode 100-1000 Da

[1036] Column: XBridge C18, 3.5μm 4.6x50mm

[1037] Temperature: 40℃

[1038] gradient:

[1039] 0.00 95％ 5％ 2.0 1.40 5％ 95％ 2.0 3.10 5％ 95％ 2.0

[1040] The results are shown in Table 1:

[1041] Table 1.

[1042]

[1043]

[1044]

[1045]

[1046]

[1047]

[1048] *Single enantiomer

[1049] Example 3: ARM-SAM-TIR SARM1 IC50 Measurement

[1050] This embodiment describes the assay of ARM-SAM-TIR NAD enzyme activity and its use in measuring the efficacy of compounds of Formula I in blocking SARM1-mediated NAD+ cleavage. The assay was optimized to characterize the efficacy of Formula I compounds in inhibiting SARM1 activity and to calculate the IC50 value for each compound. The assay utilizes the full-length SARM1, which contains ARM, SAM, and TIR domains. As demonstrated herein, expression of this fragment without its self-repressive N-terminal domain produces a constitutively active enzyme that cleaves NAD+.

[1051] Preparation of ARM-SAM-TIR lysates (STL)

[1052] NRK1-HEK 293T cells were seeded at a rate of 20 × 10⁶ cells / plate at a depth of 150 cm. 2On the plate. The next day, cells were transfected with 15 μg of ARM-SAM-TIR expression plasmid SEQ ID NO:1.

[1053]

[1054]

[1055] Supplement the culture with 1 mM NR at transfection to minimize toxicity from ARM-SAM-TIR overexpression. Forty-eight hours post-transfection, harvest cells, centrifuge at 1,000 rpm (Sorvall ST 16R centrifuge, Thermo Fisher) to pellet, and wash once with cold PBS (0.01 M phosphate-buffered saline, 0.138 M NaCl; 0.0027 M KCl; pH 7.4). Resuspend cells in a solution containing a protease inhibitor (COP). TM Cell lysates were prepared by sonication (Branson Sonifer 450, output = 3, 20 episodes of stroke) in PBS containing a mixture of protease inhibitors (Roche product #11873580001). The lysates were centrifuged (12,000 × g, 10 min at 4 °C) to remove cell debris, and the supernatant (containing ARM-SAM-TIR protein) was stored at -80 °C for later use in in vitro ARM-SAM-TIR NAD enzyme assays (see below). Protein concentration was determined by the bicinchoninic (BCA) method and used to calibrate the lysate concentrations.

[1056] ARM-SAM-TIR of Compound I IC50 measurement

[1057] Enzyme assays were performed in Dulbecco PBS buffer in 384-well polypropylene plates, with a final assay volume of 20 μL. ARM-SAM-TIR lysates at a final concentration of 5 μg / mL were pre-incubated with the corresponding compounds in 1% DMSO at the final assay concentration for 2 h at room temperature. The reaction was initiated by adding 5 μM of NAD+ at the final assay concentration as substrate. After incubation at room temperature for 2 h, the reaction was terminated with 40 μL of 7.5% trichloroacetic acid in acetonitrile. NAD+ and ADPR concentrations were analyzed using an API4000 triple quadrupole mass spectrometer (AB Sciex Framingham, MA) and a RapidFire high-throughput mass spectrometry system (Agilent Technologies, Santa Clara, CA).

[1058] The results are shown in Table 2 below. Compounds with the specified activity "A" have IC50. 50<50 nM; compounds with the specified activity "B" have IC50. 50 51-100 nM; compounds with a specified activity "C" have IC50. 50 101-500 nM; compounds with the specified activity "D" have IC50. 50 501-1000 nM; compounds with a specified activity "E" have IC50. 50 >1000nM.

[1059] Table 2.

[1060]

[1061]

[1062]

[1063] *Single enantiomer

[1064] Example 4: Axial Sudden Change Index

[1065] This embodiment illustrates an in vitro axonal degeneration assay used to characterize the compound of formula I. This assay was used to test the efficacy of the compound of formula I in preventing axonal degeneration in mouse dorsal root ganglion (DRG) hanging drop cultures.

[1066] Mouse DRG hanging drop culture: Dorsal root ganglion neurons were excised from E12.5 CD1 mice (50 ganglia per embryo) and incubated at 37°C for 15 min with 0.5% trypsin solution (Gibco) containing 0.02% EDTA. Cells were then gently pipetted and washed three times with Neurobasal medium (Gibco) containing 2% B27 (Invitrogen), 100 ng / ml 2.5SNGF (Harland Bioproducts), 1 mM 5-fluoro-2'-deoxyuridine (Sigma), penicillin, and streptomycin. Cells were then resuspended in DRG growth medium. DRG hanging drop cultures were generated by spotting 5000 cells / well into the center of each well of a 96-well tissue culture plate coated with poly-D-lysine (0.1 mg / ml; Sigma) and laminin (3 mg / ml; Invitrogen). Allow the cells to adhere to the plate in a humidified tissue culture incubator (5% CO2) for 15 min, then gently add DRG growth medium (100 mL well).

[1067] Axonal degeneration assay: Axonal degeneration was stimulated by manual axonal transecting with a scalpel blade or by chemical toxicity stimulation. After the appropriate experimental period, DRG cultures were fixed in 1% PFA + sucrose and kept in a refrigerator before imaging. Bright-field images of DRG axons and cell bodies were acquired using a 20x water immersion lens on a Phenix automated confocal microscope (PerkinElmer), and axonal quantification was performed using an internally developed protocol (Acapella, PerkinElmer).

Claims

1. A compound selected from: , and Or its pharmaceutically acceptable salt.

2. The compound of claim 1, wherein the compound is selected from: , and Or its pharmaceutically acceptable salt.

3. A pharmaceutical composition comprising the compound of claim 1 or claim 2 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers, diluents or excipients.

4. Use of the compound of claim 1 or claim 2 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment or prevention of axonal degeneration.

5. Use of the compound of claim 1 or claim 2 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of amyotrophic lateral sclerosis (ALS).

6. Use of the compound of claim 1 or claim 2 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of multiple sclerosis.

7. Use of the compound of claim 1 or claim 2 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of progressive supranuclear palsy.

Images