SARM1 inhibitors

Abstract

The present application relates to SARM1 inhibitors. The present disclosure provides compounds and methods useful for inhibiting SARM1 and / or treating and / or preventing axial degeneration. The provided SARM1 inhibitors reduce or inhibit binding of SARM1 to NAD +, and the SARM1 inhibitors bind to SARM1 within a pocket comprising one or more catalytic residues, such as the catalytic fracture of SARM1.

Description

[0001] This application is a divisional application of the patent application for "SARM1 Inhibitor" with the filing date of June 6, 2019, application number 201980052440.2.

[0002] Cross-reference to related applications

[0003] This application claims the benefit of U.S. Provisional Application No. 62 / 682,054, filed on June 7, 2018, which is incorporated herein by reference in its entirety.

[0004] Sequence Listing

[0005] This application contains a sequence listing, which is submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. The ASCII copy created on June 6, 2019, is named 2012800-0024_SL.txt and is 13,851 bytes in size. BACKGROUND OF THE INVENTION

[0006] Axonal degeneration is a hallmark of multiple neurological disorders, including peripheral neuropathy, traumatic brain injury, and neurodegenerative diseases (Gerdts et al., SARM1 activation triggers axon degeneration locally via NAD(+) destruction. Science 348 2016, pp. 453-457, which is hereby incorporated by reference in its entirety). Neurodegenerative diseases and injuries are devastating for patients and caregivers. Currently, in the United States alone, the costs associated with these diseases exceed hundreds of billions of dollars annually. Since the incidence of many of these diseases and disorders increases with age, their incidence is rapidly increasing with the changing demographics. SUMMARY OF THE INVENTION

[0007] The present disclosure provides, among other things, techniques for treating and / or preventing neurodegeneration (e.g., for reducing axonal degeneration). In some embodiments, the techniques provided inhibit SARM1.

[0008] In some embodiments, the present disclosure provides certain compounds and / or compositions that can be used in medicine, particularly for treating neurodegeneration (e.g., for reducing axonal degeneration).

[0009] In some embodiments, the present disclosure provides compounds having the structure shown in Formula I:

[0010]

[0011] or a pharmaceutically acceptable salt thereof, wherein

[0012] R 1 Selected from -CN, -NO2, -C(O)R, -S(O)2R, -CON(R)2, -S(O)2N(R)2 and -CO2R;

[0013] R 2 is-R;

[0014] R 3 Yes - (CH2) 0-2 Cy, or:

[0015] R 2 and R 3 Together with the nitrogen atom to which they are attached, they form a 4- to 7-membered saturated or partially unsaturated ring fused to Cy, or form a 4- to 7-membered saturated or partially unsaturated ring substituted by -Cy;

[0016] Cy is selected from phenyl, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, an 8- to 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8- to 10-membered bicyclic aryl ring, wherein each phenyl, heteroaryl, and aryl ring is surrounded by 0-4 R x replace;

[0017] Each R x independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R and optionally substituted C 1-6 aliphatic;

[0018] R 4 is -R; and

[0019] Each R is independently hydrogen or optionally substituted C 1-6 Aliphatic; or:

[0020] In some embodiments, two R atoms, together with the atoms to which they are attached, form a 3- to 6-membered saturated or partially unsaturated heterocyclic ring.

[0021] In some embodiments, the provided compounds have the structures of Formula I-a, I-a-i, I-a-ii, I-a-iii, I-a-iv, I-a-v, I-a-vi, I-b, I-b-i, I-b-ii, I-b-iii, I-b-iv, I-b-v, I-b-vi, I-c, I-c-i, I-c-ii, I-c-iii, I-c-iv, I-c-v, and I-c-vi as described below.

[0022] In some embodiments, the present disclosure provides compounds having the structure shown in Formula II:

[0023]

[0024] or a pharmaceutically acceptable salt thereof, wherein

[0025] R 1 is selected from -CN, -NO2, -C(O)R, -S(O)2R, -CON(R)2, -S(O)2N(R)2, -CO2R, -C(═NR)N(R)2, and a 5-membered heteroaryl ring having 2 - 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur;

[0026] R 2 is -R;

[0027] R 3 is -(CH2) 0-2 Cy, or:

[0028] R 2 and R 3 together with the nitrogen atom to which they are attached form a 4 - to 7 - membered saturated or partially unsaturated ring fused to Cy, or form a 4 - to 7 - membered saturated or partially unsaturated ring substituted with -Cy;

[0029] Cy is selected from phenyl, a 3 - to 7 - membered saturated or partially unsaturated carbocyclic ring, a 5 - to 6 - membered heteroaryl ring having 1 - 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, an 8 - to 10 - membered bicyclic heteroaryl ring having 1 - 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8 - to 10 - membered bicyclic aryl ring, wherein each phenyl, carbocyclic ring, heteroaryl ring, and aryl ring is substituted with 0 - 4 R x substituents;

[0030] Each R x is independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R, optionally substituted C 1-6an aliphatic group, an optionally substituted 5- to 6-membered saturated or partially unsaturated heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an optionally substituted 8- to 10-membered heteroaryl ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur;

[0031] R 4 is -R; and

[0032] each R is independently hydrogen, an optionally substituted C 1-6 aliphatic, an optionally substituted phenyl, and an optionally substituted 3- to 7-membered saturated or partially unsaturated carbocycle, or:

[0033] Examples of two Rs together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocycle.

[0034] In some embodiments, one or more compounds of Formula I or Formula II are provided and / or utilized in solid form (e.g., crystalline form or amorphous form).

[0035] In some embodiments, the present disclosure provides a composition comprising and / or delivering a compound of Formula I or Formula II (e.g., in the form described herein), a prodrug thereof, or an active metabolite.

[0036] In some embodiments, the present disclosure provides a composition comprising and / or delivering a compound of Formula I or Formula II. In some embodiments, such compositions are pharmaceutical compositions comprising at least one pharmaceutically acceptable carrier, diluent, or excipient.

[0037] In some embodiments, the provided SARM1 inhibitor reduces or inhibits the binding of SARM1 to NAD+. In some embodiments, the provided SARM1 inhibitor binds to SARM1 within a pocket comprising one or more catalytic residues (e.g., the catalytic cleft of SARM1).

[0038] In some embodiments, the provided compound and / or composition inhibits the activity of SARM1. Alternatively or additionally, in some embodiments, the provided compound alleviates one or more attributes of neurodegeneration. In some embodiments, the present disclosure provides a method of treating a neurodegenerative disease or disorder associated with axonal degeneration.

[0039] In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, in medical practice. In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, to treat, prevent, or ameliorate axonal degeneration (e.g., one or more of its characteristics or properties). In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, to inhibit axonal degeneration, including axonal degeneration caused by NAD+ reduction or depletion. In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, to prevent axonal degeneration distal to axonal injury.

[0040] In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, to treat one or more neurodegenerative diseases, disorders, or conditions selected from the group consisting of neuropathy or axonopathy. In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, to treat neuropathy or axonopathy associated with axonal degeneration. In some embodiments, neuropathy associated with axonal degeneration is a hereditary or congenital neuropathy or axonopathy. In some embodiments, neuropathy associated with axonal degeneration is caused by de novo or somatic mutations. In some embodiments, neuropathy associated with axonal degeneration is selected from the list included herein. In some embodiments, neuropathy or axonopathy is associated with axonal degeneration, including but not limited to Parkinson's disease, non-Parkinson's disease, Alzheimer's disease, herpes infection, diabetes, amyotrophic lateral sclerosis (ALS), demyelinating diseases, ischemia or stroke, chemical injury, thermal injury, and AIDS.

[0041] In some embodiments, a subject to whom a compound or composition described herein is administered can be or comprise a subject having or susceptible to a neurodegenerative disease, disorder, or condition. In some embodiments, the neurodegenerative disease, disorder, or condition can be or comprise 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, penetrative injury within or to the cranial cavity or body innervation regions. In some embodiments, traumatic neuronal injury is a force that causes axonal deformation, stretching, compression, or shearing.

[0042] In some embodiments, the provided method comprises administering to a patient in need a compound described herein (e.g., a compound of Formula I or Formula II). In some such embodiments, the patient is at risk of developing a condition characterized by axonal degeneration. In some embodiments, the patient has a condition characterized by axonal degeneration. In some embodiments, the patient has been diagnosed with a condition characterized by axonal degeneration.

[0043] In some embodiments, the provided method comprises administering to a population of patients in need a composition described herein. In some embodiments, the population is those individuals who are involved in activities with a high likelihood of traumatic neuronal injury. In some embodiments, the population is those athletes who are involved in contact sports or other high-risk activities.

[0044] In some embodiments, the patient is at risk of developing a neurodegenerative disorder. In some embodiments, the patient is an elderly person. In some embodiments, the patient is known to have a genetic risk factor for neurodegeneration.

[0045] In certain embodiments, the present disclosure provides compounds for use as, for example, an analytical tool, a probe in a bioassay, or a therapeutic agent according to the present disclosure. The compounds provided by the present disclosure can also be used to study the function of SARM1 in biological and pathological phenomena and for comparative evaluation of new SARM1 activity inhibitors in vitro or in vivo.

[0046] In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, as a method for inhibiting neuronal degeneration derived from a subject. In some embodiments, one or more of the compounds and / or compositions described herein can be used to inhibit the degeneration of neurons or a portion thereof cultured in vitro. In some embodiments, one or more of the compounds and / or compositions described herein can be used as a stabilizer to promote neuronal survival in vitro. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 Shows the structure of the SARM1 protein.

[0049] Figure 2 Shows that the NRK1-HEK293T stable line with NR supplementation maintains a higher NAD+ level after SARM1-TIR expression.

[0050] DEFINITIONS

[0051] Aliphatic: The term "aliphatic" refers to a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon that is completely saturated or contains one or more unsaturated units but is not aromatic (also referred to herein as "carbocyclic" or "cycloaliphatic"), which has a single point of attachment to the remainder of the molecule. Unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms. In some embodiments, "cycloaliphatic" (or "carbocyclic") refers to a monocyclic C3-C8 hydrocarbon or a bicyclic C7-C 10 hydrocarbon that is completely saturated or contains one or more unsaturated units but is not aromatic, which has a single point of attachment to the remainder of the molecule. Suitable aliphatic groups include, but are not limited to, straight-chain or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl, and their hybrids.

[0052] Alkyl: The term "alkyl", used alone or as part of a larger moiety, 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 an optionally substituted saturated ring system having from about 3 to about 10 ring carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0053] 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, "alkylene chain" is polymethylene, i.e., -(CH2) n -, where n is a positive integer, such as 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. An optionally substituted alkylene chain is a polymethylene in which 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 the present specification. It should be understood that two substituents of an alkylene group can together form a ring system. In certain embodiments, the two substituents can together form a 3- to 7-membered ring. The substituents can be on the same or different atoms.

[0054] Alkenyl: The term "alkenyl", used alone or as part of a larger moiety, refers to an optionally substituted straight-chain, branched-chain, 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 from about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include cyclopentyl, cyclohexenyl, and cycloheptenyl.

[0055] Alkynyl: The term "alkynyl", used alone or as part of a larger moiety, refers to an optionally substituted straight-chain or branched-chain 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.

[0056] Aryl: The term "aryl" refers to monocyclic and bicyclic ring systems having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. The term "aryl" may be used interchangeably with the term "aryl ring". In certain embodiments of the present invention, "aryl" refers to an aromatic ring system including, but not limited to, phenyl, biphenyl, naphthyl, anthracenyl, etc., which may bear one or more substituents. As used herein, groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimido, naphthalimido, phenanthridinyl, or tetrahydronaphthyl, etc., are also included within the scope of the term "aryl".

[0057] Binding: It should be understood that the term "binding" as used herein generally refers to non-covalent association between or among two or more entities. "Direct" binding involves physical contact between entities or moieties; 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 situations, including studying the interacting entities or moieties in isolation or in a more complex system (e.g., when covalently or otherwise associated with a carrier entity and / or in a biological system or cell).

[0058] Biological sample: As used herein, the term "biological sample" generally 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 comprises an organism, such as an animal or a human. In some embodiments, the biological sample is or comprises biological tissue or fluid. In some embodiments, the biological sample can be or comprise bone marrow; blood; blood cells; ascites; tissue or fine needle biopsy samples; cell-containing body fluids; cell-free nucleic acids; 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 lavage or bronchoalveolar lavage; aspirates; scrapings; bone marrow specimens; tissue biopsy specimens; surgical specimens; feces, other body fluids, secretions, and / or excretions; and / or cells obtained therefrom, etc. In some embodiments, the biological sample is or comprises cells obtained from an individual. In some embodiments, the cells obtained are or include cells of the individual from whom the sample was obtained. In some embodiments, the sample is an "original sample" obtained directly from the source of interest by any suitable means. For example, in some embodiments, the original biological sample is obtained by a method selected from the group consisting of biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of body fluids (e.g., blood, lymph, feces, etc.), etc. In some embodiments, as will be apparent from the context, the term "sample" refers to a preparation obtained by processing the original sample (e.g., by removing one or more components of the original sample and / or by adding one or more reagents thereto). For example, filtration using a semipermeable membrane. Such a "processed sample" can contain, for example, nucleic acids or proteins extracted from the sample or obtained by techniques such as amplification or reverse transcription of mRNA, separation and / or purification of certain components, etc., of the original sample.

[0059] Biomarker: The term "biomarker" is used herein to refer to an entity, event, or characteristic whose presence, level, degree, type, and / or form is associated with a particular biological event or state of interest, and thus it is considered a "marker" of that event or state. By way of example only, in some embodiments, a biomarker can be or include a marker of a particular disease state, or a marker of the likelihood that a particular disease, disorder, or condition may develop, occur, or recur. In some embodiments, a biomarker can be or include a marker of a particular disease or its treatment outcome or likelihood. Thus, in some embodiments, a biomarker is a predictor of a relevant biological event or state of interest, in some embodiments, a biomarker is a prognosis of a relevant biological event or state of interest, and in some embodiments, a biomarker is a diagnosis of a relevant biological event or state of interest. A biomarker can be or include entities of any chemical class and can be or include combinations of entities. For example, in some embodiments, a biomarker can be or include nucleic acids, polypeptides, 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, a biomarker is detected extracellularly (e.g., is secreted or otherwise produced or present extracellularly, such as in a body fluid, such as blood, urine, tears, saliva, cerebrospinal fluid, etc.). In some embodiments, a biomarker can be or include genetic or epigenetic features. In some embodiments, a biomarker can be or include gene expression features.

[0060] In some embodiments, a biomarker can 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, a biomarker can be or include a marker of neurodegeneration, its treatment outcome, or likelihood. Thus, in some embodiments, a biomarker is a predictor of a neurodegenerative disease, disorder, or condition, in some embodiments, a biomarker is a prognosis of a neurodegenerative disease, disorder, or condition, and in some embodiments, a biomarker is a diagnosis of a neurodegenerative disease, disorder, or condition. In some embodiments, changes in biomarker levels can be detected by cerebrospinal fluid (CSF), plasma, and / or serum.

[0061] In some embodiments, neurodegeneration can be evaluated, for example, by detecting an increase and / or decrease in the concentration of neurofilament light chain (NF-L) and / or neurofilament heavy chain (NF-H) contained in the cerebrospinal fluid of a subject. In some embodiments, the occurrence and / or progression of neurodegeneration can be evaluated by positron emission tomography (PET) with a synaptic vesicle glycoprotein 2a (SV2A) ligand. In some embodiments, detectable changes in the levels of constitutive NAD and / or cADPR in neurons can be used to evaluate neurodegeneration.

[0062] 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, neurogranin, neuron-specific enolase (NSE), soluble amyloid precursor protein (sAPP) α, sAPPβ, soluble triggering receptor expressed on myeloid cells (sTREM) 2, phosphorylated tau, and / or total tau. In some embodiments, an increase in cytokines and / or chemokines, including but not limited to Ccl2, Ccl7, Ccl12, Csf1, and / or Il6, can be used as a biomarker of neurodegeneration.

[0063] Vehicle: As used herein, the term "vehicle" refers to a diluent, adjuvant, excipient, or medium with which a composition is administered. In some exemplary embodiments, the vehicle can include sterile liquids, such as water and oils, including oils of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. In some embodiments, the vehicle is or includes one or more solid components.

[0064] Combination Therapy: As used herein, the term "combination therapy" refers to those situations where 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 can be administered simultaneously; in some embodiments, such regimens can be administered sequentially (e.g., all "doses" 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 a combination therapy can involve administering one or more agents or moieties to a subject who is receiving other agents or moieties in the combination. For clarity, combination therapy does not require that the individual agents be administered together (or even necessarily simultaneously) in a single composition, although in some embodiments, two or more agents or their active portions can 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).

[0065] Composition: Those skilled in the art will appreciate that the term "composition" can be used to refer to a discrete physical entity that contains one or more specified components. Generally, unless otherwise specified, a composition can be in any form, such as a gas, gel, liquid, solid, etc.

[0066] Domain: As used herein, the term "domain" refers to a segment or portion of an entity. In some embodiments, a "domain" is associated with specific structural and / or functional characteristics of the entity, and thus, when physically separated from the remainder of its parent entity, it substantially or completely retains the specific structural and / or functional characteristics. Alternatively or additionally, a domain can be or include a portion of an entity that, when separated from the (parent) entity and attached to a different (receptor) entity, substantially retains and / or confers upon the receptor entity one or more structural and / or functional characteristics that are characteristic of it in the parent 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 segment of a polypeptide; in some such embodiments, the domain is characterized by specific structural elements (e.g., a specific amino acid sequence or sequence motif, an α-helical characteristic, a β-sheet characteristic, a coiled-coil characteristic, a random coil characteristic, etc.), and / or by specific functional characteristics (e.g., binding activity, enzyme activity, folding activity, signaling activity, etc.).

[0067] Dosage form or unit dosage form: Those skilled in the art should understand that the term "dosage form" can be used to refer to a physically discrete unit of an active agent (e.g., a therapeutic agent or a diagnostic agent) for administration to a subject. Generally, each such unit contains a predetermined amount of the active agent. In some embodiments, such an amount is a unit dose (or a fraction thereof) suitable for administration according to a dosing regimen (i.e., a therapeutic dosing regimen) that has been determined to be associated with a desired or beneficial outcome when administered to the relevant population. Those of ordinary skill in the art 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.

[0068] Dosing regimen or treatment regimen: Those skilled in the art should understand that the terms "dosing regimen" and "treatment regimen" can be used to refer to a set of unit doses (usually more than one) administered individually to a subject, and the unit doses are usually separated by time periods. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses. In some embodiments, the dosing regimen includes multiple doses, where each dose is separated from the other doses in time. In some embodiments, the individual doses are separated by the same length of time period; in some embodiments, the dosing regimen includes multiple doses and at least two different time periods separating the individual doses. In some embodiments, all the doses within the dosing regimen have the same amount of the unit dose. In some embodiments, the different doses within the dosing regimen have different amounts. In some embodiments, the dosing regimen contains a first dose in an amount of a first dose, followed by one or more additional doses in an amount of a second dose different from the first dose. In some embodiments, the dosing regimen contains a first dose in an amount of a first dose, followed by one or more additional doses in an amount of a second dose the same as the amount of the first dose. In some embodiments, when administered to the relevant population, the dosing regimen is associated with a desired or beneficial outcome (i.e., is a therapeutic dosing regimen).

[0069] Excipient: As used herein, it refers to a non-therapeutic agent that may be included in a pharmaceutical composition, for example, to provide or facilitate the desired consistency or stabilizing effect. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene, ethylene glycol, water, ethanol, etc.

[0070] Heteroaryl: The terms "heteroaryl" and "hetero-" as used alone or as part of a larger moiety (such as "heteroalkyl" or "heteroalkoxy") refer to groups having 5 to 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 carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur and includes any oxidized form of nitrogen or sulfur and any quaternized form of basic nitrogen. Heteroaryl includes, but is not limited to, thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. The terms "heteroaryl" and "hetero-" as used herein also include groups in which a heteroaromatic ring is fused to one or more aryl rings, cycloaliphatic rings, or heterocyclic rings, where the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuryl, dibenzofuryl, indazolyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl can be monocyclic or bicyclic. The term "heteroaryl" can be used interchangeably with the terms "heteroaryl ring", "heteroaryl group", or "heteroaromatic", any of which includes a ring optionally substituted. The term "heteroalkyl" refers to an alkyl group substituted with a heteroaryl group, where the alkyl and heteroaryl moieties are independently optionally substituted.

[0071] Heterocycle: As used herein, the terms "heterocycle", "heterocyclyl", "heterocyclic radical", and "heterocyclic ring" are used interchangeably and refer to a stable 3- to 8-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is saturated or partially unsaturated and has one or more, such as one to four, heteroatoms in addition to carbon atoms, as defined above. When used in reference to the ring atoms of a heterocycle, the term "nitrogen" includes substituted nitrogen. For example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, and nitrogen, nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR +(such as in N - substituted pyrrolidinyl). The heterocycle can be attached to its side group at any heteroatom or carbon atom, resulting in a stable structure, and any ring atom can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxolanyl, dioxolyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and thiomorpholinyl. The heterocyclic group can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. The term "heterocyclic alkyl" refers to an alkyl group substituted with a heterocyclic group, wherein the alkyl and heterocyclic moieties are independently optionally substituted. In addition, the heterocycle also includes groups in which the heterocycle is fused to one or more aromatic rings.

[0072] Inhibitor: As used herein, the term "inhibitor" refers to an entity, condition, or event whose presence, level, or degree is associated with a decrease in the level or activity of a target. In some embodiments, the inhibitor can act directly (in which case it directly exerts an influence on its target, e.g., by binding to the target); in some embodiments, the inhibitor can act indirectly (in which case it exerts an influence by interacting with a modulator of the target and / or otherwise altering the modulator of the target such that the level and / or activity of the target is decreased). In some embodiments, the inhibitor is an inhibitor whose presence or level is associated with a decrease in the level or activity of a target relative to a specific reference level or activity (e.g., the level or activity observed under appropriate reference conditions, such as in the presence of a known inhibitor, or in the absence of the inhibitor being discussed, etc.).

[0073] Neurodegeneration: As used herein, the term "neurodegeneration" refers to a decrease in one or more characteristics, structures, or properties of neurons or neuronal tissue. In some embodiments, neurodegeneration is observed as a pathological decrease in an organism. Those skilled in the art will appreciate that neurodegeneration is associated with certain diseases, disorders, and conditions, including those that affect humans. In some embodiments, neurodegeneration can be transient (e.g., sometimes occurring in association with certain infections and / or chemical or mechanical disruptions); in some embodiments, neurodegeneration can be chronic and / or progressive (e.g., often associated with certain diseases, disorders, or conditions such as, but not limited to, Parkinson's disease, amyotrophic lateral sclerosis, multiple sclerosis, Huntington disease, or Alzheimer's disease). In some embodiments, neurodegeneration can be evaluated, for example, by detecting an increase in a biomarker associated with neurodegeneration in a subject. In some embodiments, neurodegeneration can be evaluated, for example, by detecting a decrease in a biomarker associated with neurodegeneration in a subject. Alternatively or additionally, in some embodiments, neurodegeneration can be evaluated by magnetic resonance imaging (MRI), cerebrospinal fluid containing biomarkers, or other biomarkers observed in a patient. In some embodiments, neurodegeneration is defined as a score on the Mini-Mental State Examination of less than 24. In some embodiments, neurodegeneration refers to the loss of synapses. In some embodiments, neurodegeneration refers to a reduction in neural tissue associated with a traumatic injury (e.g., exposure to an external force that disrupts 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 neural tissue.

[0074] Oral: The phrases "oral administration" and "administered orally" as used herein have the meanings understood in the art and refer to the administration of a compound or composition through the mouth.

[0075] Parenteral: The phrases "parenteral administration" and "administered parenterally" as used herein have the meanings understood in the art and refer to a mode of administration other than enteral and topical administration (usually by injection) and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0076] Partially unsaturated: As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond between ring atoms. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic (e.g., aryl or heteroaryl) moieties as defined herein.

[0077] Patient: As used herein, the term "patient" refers to any organism to which the provided composition is administered or can be administered, for example, for experimental, diagnostic, prophylactic, 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 disorders or conditions. In some embodiments, the patient exhibits one or more symptoms of a disorder or condition. In some embodiments, the patient has been diagnosed with one or more disorders or conditions. In some embodiments, the patient is receiving or has received certain therapies for diagnosing and / or treating a disease, disorder, or condition.

[0078] 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 an amount suitable for administration in a unit dose of a therapeutic or dosing regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to the relevant population. In some embodiments, the pharmaceutical composition can be specifically formulated for administration in solid or liquid form, including pharmaceutical compositions suitable for: oral administration, such as drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., tablets targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes applied 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 applied to the skin, lung, or oral cavity as a cream, ointment, or controlled release patch or spray; intravaginally or rectally, such as as a pessary, cream, or foam; sublingually; ophthalmically; transdermally; or via the nasal, pulmonary, and other mucosal surfaces.

[0079] Pharmaceutically acceptable: As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the scope of reasonable medical judgment, are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, and are commensurate with a reasonable benefit / risk ratio.

[0080] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material involved 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 being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; 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 glycerin, sorbitol, mannitol, and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; pH buffering solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic compatible substances employed in pharmaceutical formulations.

[0081] Pharmaceutically acceptable salts: The term "pharmaceutically acceptable salts" as used herein refers to salts of such compounds that are suitable for use in a pharmaceutical context, i.e., salts that are suitable for contact with the tissues of humans and lower animals within the scope of reasonable medical judgment, without undue toxicity, irritation, allergic response, etc., and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in detail by S.M. Berge et al. in Journal of 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 the reaction of an amino group with an inorganic acid or an organic acid, inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or salts formed by using other methods used in the art, such as ion exchange. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecyl sulfates, ethanesulfonates, formates, fumarates, glucoheptanoates, glycerophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxyethanesulfonates, lactates, lactobionates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, etc. Representative alkali metal salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. In some embodiments, where appropriate, pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions, counterions such as halide ions, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl groups having 1 to 6 carbon atoms, sulfonate, and arylsulfonate.

[0082] Prevent or prevention: As used herein, the terms "prevent" or "prevention" when used in connection with the occurrence of a disease, disorder, and / or condition refer to reducing the risk of developing a disease, disorder, and / or condition and / or delaying the onset of one or more characteristics 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 predetermined period of time.

[0083] Specificity: The term "specificity", as used herein when referring to an agent having activity, is understood by those skilled in the art to mean that the agent discriminates between a potential target entity or state. For example, in some embodiments, an agent is said to "specifically" bind to a target if it preferentially binds to its 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., epitope, cleft, binding site). It should be understood that specificity need not be absolute. In some embodiments, specificity can be evaluated relative to the specificity of the binder for 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, no binding to a competing alternative target is detected for the agent or entity under conditions where it binds to its target entity. In some embodiments, the binder binds to its target entity with a higher binding rate, a lower dissociation rate, an increased affinity, a reduced dissociation, and / or an increased stability compared to a competing alternative target.

[0084] Subject: As used herein, the term "subject" refers to an organism, typically a mammal (e.g., a human, including in some embodiments a pre-birth human form). In some embodiments, the subject has a relevant disease, disorder, or condition. In some embodiments, the subject is predisposed to a disease, disorder, or condition. In some embodiments, the subject exhibits one or more symptoms or features of a disease, disorder, or condition. In some embodiments, the subject does not exhibit any symptoms or features of a disease, disorder, or condition. In some embodiments, the subject is a human having one or more characteristics of susceptibility or risk for a disease, disorder, or condition. In some embodiments, the subject is a patient. In some embodiments, the subject is an individual who is being and / or has been diagnosed and / or treated.

[0085] Substituted or optionally substituted: As described herein, the compounds of the present invention may contain moieties that are "optionally substituted". In general, the term "substituted", whether or not preceded by the term "optionally", means that one or more hydrogens of the designated moiety are replaced by a suitable substituent. "Substituted" applies to one or more hydrogens that are explicitly or implicitly present in the structure (e.g., at least refers to and at least refers to )。Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure can be substituted by more than one substituent selected from the designated groups, the substituents may be the same or different at each position. Preferred combinations of substituents contemplated by the present invention are those that result in the formation of stable or chemically viable compounds. As used herein, the term "stable" means that a compound remains substantially unchanged when subjected to conditions that permit its production, detection, and in some embodiments, its recovery, purification, and use for one or more of the purposes disclosed herein.

[0086] Suitable monovalent substituents on the substitutable carbon atoms of an "optionally substituted" group are independently halogen; -(CH2) 0-4 R o ; -(CH2) 0-4 OR o ; -O(CH2) 0-4 R o ; -O-(CH2) 0-4 C(O)OR o ; -(CH2) 0-4 CH(OR o )2; -(CH2) 0- 4SR o ; -(CH2) 0-4 Ph, which may be substituted by R o ; -(CH2) 0-4 O(CH2) 0-1 Ph, which may be substituted by R o ; -CH=CHPh, which may be substituted by R o ; -(CH2) 0-4 O(CH2) 0-1 -pyridyl, which may be substituted by R o ; -NO2; -CN; -N3; -(CH2) 0-4 N(R o )2; -(CH2) 0-4 N(R o )C(O)R o ; -N(R o )C(S)R o ; -(CH2) 0-4 N(R o )C(O)NR o 2; -N(R o )C(S)NR o 2; -(CH2) 0-4 N(R o )C(O)OR o ; -N(Ro )N(R o )C(O)R o ;-N(R o )N(R o )C(O)NR o 2;-N(R o )N(R o )C(O)OR o ;-(CH2) 0-4 C(O)R o ;-C(S)R o ;-(CH2) 0-4 C(O)OR o ;-(CH2) 0-4 C(O)SR o ;-(CH2) 0-4 C(O)OSiR o 3;-(CH2) 0-4 OC(O)R o ;-OC(O)(CH2) 0-4 SR o ;-(CH2) 0-4 SC(O)R o ;-(CH2) 0-4 C(O)NR o 2;-C(S)NR o 2;-C(S)SR o ;-SC(S)SR o ;-(CH2) 0- 4OC(O)NR o 2;-C(O)N(OR o )R o ;-C(O)C(O)R o ;-C(O)CH2C(O)R o ;-C(NOR o )R o ;-(CH2) 0-4 SSR o ;-(CH2) 0-4 S(O)2R o ;-(CH2) 0-4 S(O)2OR o ;-(CH2) 0-4 OS(O)2R o ;-S(O)2NR o 2;-(CH2) 0-4 S(O)R o ;-N(R o )S(O)2NR o 2;-N(R o)S(O)2R o ;-N(OR o )R o ;-C(NH)NR o 2;-P(O)2R o ;-P(O)R o 2;-OP(O)R o 2;-OP(O)(OR o )2;SiR o 3;-(C 1-4 linear or branched alkylene)O-N(R o )2;or-(C 1-4 linear or branched alkylene)C(O)O-N(R o )2, where each R o may be substituted as defined below and is independently hydrogen, C 1-6 aliphatic, -CH2Ph, -O(CH2) 0- 1Ph, -CH2-(5- to 6-membered heteroaryl ring), a 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or an 8- to 10-membered bicyclic aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definition, two independently occurring R o together with their intervening atom form a 3- to 12-membered saturated, partially unsaturated or aryl monocyclic, bicyclic or bridged ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

[0087] R o (or the ring formed by two independently occurring R o together with their intervening atom) suitable monovalent substituents on are independently halogen, -(CH2) 0-2 R · , -(haloalkyl R · ), -(CH2) 0-2 OH, -(CH2) 0-2 OR · , -(CH2) 0-2 CH(OR · )2, -O(haloalkyl R · ), -CN, -N3, -(CH2) 0-2 C(O)R · , -(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR · , -(CH2) 0-2 SR · , -(CH2) 0-2SH, -(CH2) 0-2 NH2, -(CH2) 0-2 NHR · , -(CH2) 0-2 NR · 2, -NO2, -SiR · 3, -OSiR · 3, -C(O)SR · , -(C 1-4 linear or branched alkylene)C(O)OR · or -SSR · , wherein each R ● is unsubstituted, or where preceded by "halo" is substituted by one or more halogens only, and is independently selected from C 1-4 aliphatic, -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 or sulfur. Suitable divalent substituents on the saturated carbon atoms of R o include =O and =S.

[0088] Suitable divalent substituents on the saturated carbon atoms of a group that is "optionally substituted" include the following: =O ("oxo"), =S, =NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2)) 2- 3O- or -S(C(R * 2)) 2-3 S-, wherein each independently occurring R * is selected from hydrogen, C 1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents bonded to an ortho-substitutable carbon of a group that is "optionally substituted" include: -O(CR * 2) 2-3 O-, wherein each independently occurring R * is selected from hydrogen, C 1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0089] R *Suitable substituents on the aliphatic group include halogen, -R ● , -(haloalkyl R · ), -OH, -OR · , -O(haloalkyl R · ), -CN, -C(O)OH, -C(O)OR · , -NH2, -NHR · , -NR · 2 or -NO2, where each R ( is unsubstituted, or where preceded by "halo" is substituted only by one or more halogens, and is independently C 1-4 aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or a 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0090] Suitable substituents on the nitrogen of a group which is "optionally substituted" include

[0091] or where each is independently hydrogen, C 1-6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definition, two independently occurring together with their intervening atom form an unsubstituted 3- to 12-membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0092] Suitable substituents on the aliphatic group of ● , -(haloalkyl R ● ), -OH, -OR ● , -O(haloalkyl R ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2 or -NO2, where each R ● is unsubstituted, or where preceded by "halo" is substituted only by one or more halogens, and is independently C 1-4 aliphatic, -CH(2)Ph, -O(CH2) 0- 1Ph or a 5- to 6-membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0093] Therapeutic agent: As used herein, the phrase "therapeutic agent" generally refers to any agent that elicits a desired pharmacological effect when administered to a living organism. In some embodiments, an agent is considered a therapeutic agent if it shows a statistically significant effect in an appropriate population. In some embodiments, the appropriate population can be a population of model organisms. In some embodiments, the appropriate population can be defined by various criteria, such as a certain age group, gender, genetic background, pre-existing clinical condition, etc. In some embodiments, a therapeutic agent is a substance that can be used to alleviate, improve, relieve, inhibit, prevent one or more symptoms or characteristics of a disease, disorder, and / or condition, delay its onset, reduce its severity, and / or reduce its incidence. In some embodiments, "therapeutic agent" refers to an agent that has been or needs to be approved by a government agency for sale for administration to humans. In some embodiments, "therapeutic agent" is an agent that requires a medical prescription for administration to humans.

[0094] Treatment: As used herein, the terms "treat", "treatment", or "treating" refer to any method used to partially or completely alleviate, improve, relieve, inhibit, prevent one or more symptoms or characteristics of a disease, disorder, and / or condition, delay its onset, reduce its severity, and / or reduce its incidence. Treatment can be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition. In some embodiments, treatment can be administered to a subject who exhibits only early signs of a disease, disorder, and / or condition, for example, for the purpose of reducing the risk of developing a pathology associated with the disease, disorder, and / or condition. Detailed embodiments

[0095] Programmed axonal degeneration and SARM1

[0096] Axonal degeneration is a major pathological feature of neurological diseases, such as 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 intrinsic self-destruction program that is distinct from traditional cell death pathways, such as apoptosis, which is known as Wallerian degeneration. (Gerdts, J. et al., Neuron, 2016, 89, 449-460; Whitmore, A. V. et al., Cell Death Differ., 2003, 10, 260-261). In Wallerian degeneration, the peripheral nerve undergoes selective decomposition in the distal axonal segment of the injury, while the proximal axonal segment and cell body remain intact. This degeneration is characterized by, first, the depletion of nicotinamide mononucleotide adenylyltransferase (NMNAT), followed by the loss of nicotinamide adenine dinucleotide (NAD+), the loss of adenosine triphosphate (ATP), the proteolysis of neurofilaments, and finally axonal degeneration occurring approximately 8 to 24 hours after injury. (Gerdts, J. et al., Neuron, 2016, 89, 449-460).

[0097] NAD+ is a ubiquitous metabolite that plays a key 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). The homeostatic regulation of NAD+ levels is also responsible for maintaining axonal stability and integrity. Thus, manipulations that increase the axonal localization of NMNAT1 confer axonal protection (Babetto et al., Cell Rep., 2010, 3, 1422-1429; Sasaki et al., J. Neurosci., 2009).

[0098] In a genome-wide RNAi screen of primary mouse neurons, SARM1 (Sterile Alpha and TIR motif-containing 1) was identified, and knockdown of SARM1 confers long-lasting protection of sensory neurons against injury-induced axonal degeneration (Gerdts et al., The Journal of Neuroscience, 2013, 33, 13569-13580). SARM1 belongs to the family of cytoplasmic adaptor proteins but is unique among its members in that it is the most evolutionarily ancient adaptor, paradoxically inhibits TLR signaling, and has been identified as a central executor of the injury-induced axon death pathway (O'Neill, L.A. and Bowie, A.G., Nat. Rev. Immunol., 2007, 7, 353-364; Osterloh, J.M. et al., Science, 2012, 337, 481-484; Gerdts, J. et al., The Journal of Neuroscience 33, 2013, 13569-13580). Activation of SARM1 by axonal injury or forced dimerization of the SARM1-TIR domain promotes rapid and catastrophic depletion of nicotinamide adenine dinucleotide (NAD+), which is soon followed by axonal degeneration, thus highlighting the central role of NAD+ homeostasis in axonal integrity. (Gerdts, J. et al., Science, 2015, 348, 453-457). SARM1 is required for this injury-induced NAD+ depletion both in vivo and in vitro, and activation of SARM1 locally triggers axonal degeneration via the destruction of NAD(+) (Gerdts et al., Science, 2015 348, 452-457; Sasaki et al., J. Biol. Chem. 2015, 290, 17228-17238; both are hereby incorporated by reference in their entirety).

[0099] From loss-of-function studies, it is clear that SARM1 acts as a central executor of the axonal degeneration pathway after injury. Genetic knockout of SARM1 can preserve axons for up to 14 days after nerve transection (Osterloh, J.M. et al., Science, 2012, 337, 481-484; Gerdts, J. et al., The Journal of Neuroscience, 2013, 33, 13569-13580), and also improve functional outcomes after traumatic brain injury in mice (Henninger, N. et al., Brain, 2016, 139, 1094-1105). In addition to its role in direct axonal injury, SARM1 has been observed to be required for axonal degeneration in chemotherapy-induced peripheral neuropathy. Deletion of SARM1 blocks chemotherapy-induced peripheral neuropathy, both inhibiting axonal degeneration and the increased pain sensitivity that develops after treatment with the chemotherapy vincristine (Geisler et al., Brain, 2016, 139, 3092-3108).

[0100] SARM1 contains multiple conserved motifs, including an SAM domain, ARM / HEAT motifs, and a TIR domain ( Figure 1 ), which mediate oligomerization and protein-protein interactions (O'Neill, L.A. and Bowie, A.G., Nature Reviews Immunology, 2007, 7, 353-364; Tewari, R. et al., Trends in Cell Biology, 2010, 20, 470-481; Qiao, F. and Bowie, J.U., Sci. STKE, 2005, re7, 2005). TIR domains are typically found in signaling proteins that function in innate immune pathways, where they act as scaffolds for protein complexes (O'Neill, L.A. and Bowie, A.G., Nature Reviews Immunology, 2007, 7, 353-364). Interestingly, dimerization of the SARM1-TIR domain is sufficient to induce axonal degeneration and rapidly trigger the degradation of NAD+ by acting as an NAD+ lyase (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 established NADase activity, much effort has been made to identify agents that can modulate SARM1 and potentially act as useful therapeutics, for example, to prevent the development of neurodegenerative diseases, including peripheral neuropathy, traumatic brain injury, and / or neurodegenerative diseases.

[0101] Among other things, the present disclosure provides certain compounds and / or compositions that act as SARM1 inhibitors (e.g., as SARM1 inhibitors), and related technologies.

[0102] Compound

[0103] In some embodiments, the present disclosure provides a compound of formula I:

[0104]

[0105] or a pharmaceutically acceptable salt thereof, wherein:

[0106] R 1 is selected from -CN, -NO2, -C(O)R, -S(O)2R, -CON(R)2, -S(O)2N(R)2, and -CO2R;

[0107] R 2 is -R;

[0108] R 3 is -(CH2) 0-2 Cy, or:

[0109] R 2 and R 3 together with the nitrogen atom to which they are attached form a 4- to 7-membered saturated or partially unsaturated ring fused to Cy, or form a 4- to 7-membered saturated or partially unsaturated ring substituted with -Cy;

[0110] Cy is selected from phenyl, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, an 8- to 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8- to 10-membered bicyclic aryl ring, wherein each phenyl, heteroaryl ring, and aryl ring is substituted with 0-4 R x substituents;

[0111] Each R x is independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R, and optionally substituted C 1-6 aliphatic;

[0112] R 4 is -R; and

[0113] Each R is independently hydrogen or optionally substituted C 1-6 aliphatic; or:

[0114] Two instances of R, together with the atoms to which they are attached, form a 3- to 6-membered saturated or partially unsaturated heterocycle.

[0115] In some embodiments, the present disclosure provides a compound having a structure represented by Formula II:

[0116]

[0117] or a pharmaceutically acceptable salt thereof, wherein

[0118] R 1 is selected from -CN, -NO2, -C(O)R, -S(O)2R, -CON(R)2, -S(O)2N(R)2, -CO2R, -C(=NR)N(R)2, and a 5-membered heteroaryl ring having 2-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur;

[0119] R 2 is -R;

[0120] R 3 is -(CH2) 0-2 Cy, or:

[0121] R 2 and R 3 together with the nitrogen atom to which they are attached form a 4- to 7-membered saturated or partially unsaturated ring fused to Cy, or form a 4- to 7-membered saturated or partially unsaturated ring substituted with -Cy;

[0122] Cy is selected from phenyl, a 3- to 7-membered saturated or partially unsaturated carbocyclic ring, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, an 8- to 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8- to 10-membered bicyclic aryl ring, wherein each phenyl, carbocyclic ring, heteroaryl ring, and aryl ring is substituted with 0-4 R x substituents;

[0123] Each R x is independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R, an optionally substituted C 1-6 aliphatic group, an optionally substituted 5- to 6-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an optionally substituted 8- to 10-membered heteroaryl ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur;

[0124] R 4 is -R; and

[0125] Each R is independently hydrogen, optionally substituted C 1-6 aliphatic, optionally substituted phenyl, or optionally substituted 3- to 7-membered saturated or partially unsaturated carbocyclic ring, or:

[0126] Examples of two Rs together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocyclic ring.

[0127] As generally defined above, R 1 is selected from -CN, -NO2, -C(O)R, -S(O)2R, -CON(R)2, -S(O)2N(R)2 and -CO2R. In some embodiments, R 1 is selected from -CN, -C(O)N(R)2 and -CO2R. In some embodiments, R 1 is -CN. In some embodiments, R 1 is -CON(R)2. In some such embodiments, each R is independently selected from hydrogen and C 1-6 aliphatic. In some embodiments, R 1 is -CON(R)2, wherein each R is independently selected from hydrogen and optionally substituted C 1-6 aliphatic. In some embodiments, R 1 is In some embodiments, R 1 is -CON(R)2, wherein each R is independently selected from hydrogen and C 1-6 alkyl. In some embodiments, R 1 is -CON(R)2, wherein each R is independently selected from hydrogen and -CH3. In some embodiments, R 1 is -CONH2. In some embodiments, R 1 is -CON(CH3)2. In some embodiments, R 1 is In some embodiments, R 1 is -CON(R)2, wherein each R is independently selected from C 1-6 alkyl and optionally substituted phenyl. In some embodiments, R 1 is

[0128] In some embodiments, R 1 is -CON(R)2, wherein R is optionally substituted C 1-6 aliphatic, and examples of two Rs together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur. In some embodiments, R 1 is In some embodiments, R 1 is -CON(R)2, where R is optionally substituted C 1-6 aliphatic, and two instances of R together with the atoms to which they are attached form an 8- to 10-membered bridged bicyclic saturated or partially unsaturated heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R 1 is

[0129] In some embodiments, R 1 is -CO2R. In some such embodiments, R is selected from hydrogen and C 1-6 aliphatic. In some embodiments, R 1 is -CO2R, where R is selected from hydrogen and C 1-6 alkyl. In some embodiments, R 1 is -CO2R, where R is selected from hydrogen and -CH3. In some embodiments, R 1 is -CO2H. In some embodiments, R 1 is -CO2CH3. In some embodiments, R 1 is -NO2. In some embodiments, R 1 is -C(O)R. In some embodiments, R 1 is -S(O)2R. In some embodiments, R 1 is -S(O)2N(R)2.

[0130] In some embodiments of Formula II, R 1 is -C(=NR)N(R)2. In some embodiments of Formula II, R 1 is -C(=NR)N(R)2, where R is selected from hydrogen and optionally substituted C 1-6 aliphatic. In some embodiments of Formula II, R 1 is -C(=NR)N(R)2, where R is selected from hydrogen and -CH3. In some embodiments of Formula II, R 1 is -C(=NH)NHCH3. In some embodiments, R 1 is -C(=NH)N(CH3)2. In some embodiments of Formula II, R 1 is -C(=NR)N(R)2, where R is selected from hydrogen and C 0-4 substituted by -(CH2) o OR 1-6 aliphatic. In some embodiments of Formula II, R 1 is -C(=NR)N(R)2, where R is selected from hydrogen and C substituted by -OH1-6 Aliphatic. In some embodiments of Formula II, R 1 is In some embodiments of Formula II, R 1 is -C(=NR)N(R)2, where R is selected from H, -CH3, and C o substituted with -OR 1-6 Aliphatic. In some embodiments of Formula II, R 1 is

[0131] In some embodiments of Formula II, R 1 is -C(=NH)N(R)2, where R is optionally substituted C 1-6 Aliphatic, and two instances of R together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments of Formula II, R 1 is

[0132] In some embodiments of Formula II, R 1 is a 5-membered heteroaryl ring having 2 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments of Formula II, R 1 is a 5-membered heteroaryl ring having 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R 1 is

[0133] In some embodiments, R 1 is selected from -CN,

[0134]

[0135] As generally defined above, R 2 is -R. In some such embodiments, -R is hydrogen. Thus, in some embodiments, R 2 is -H. In some embodiments, R 2 is -R, where -R is optionally substituted C 1-6 Aliphatic. In some embodiments, R 2 is -R, where -R is C[[ID=�7]] 1-6 Aliphatic. In some embodiments, R 2 is -C 1-6 Alkyl. In some such embodiments, R 2 is -CH3.

[0136] As generally defined above, R 3is -(CH2) 0-2 Cy. In some embodiments, R 3 is -Cy. In some embodiments, R 3 is -CH2-Cy. In some embodiments, R 3 is -(CH2)2-Cy.

[0137] As generally defined above, Cy is selected from phenyl, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, an 8- to 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8- to 10-membered bicyclic aryl ring, wherein each phenyl, heteroaryl ring, and aryl ring is substituted with 0-4 R x substituents.

[0138] In some embodiments, Cy is phenyl. In some embodiments, Cy is phenyl substituted with 1 R x substituent. In some embodiments, Cy is phenyl substituted with 2 R x substituents. In some embodiments, Cy is selected from

[0139]

[0140] In some embodiments, Cy is a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Cy is a 5-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Cy is a 6-membered heteroaryl ring having 1-3 nitrogen atoms. In some embodiments, Cy is a 6-membered heteroaryl ring having 1-2 nitrogen atoms. In some such embodiments, Cy is substituted with 1 R x substituent. In some embodiments, Cy is pyridyl. In some such embodiments, Cy is pyrimidin-2-yl, pyrimidin-3-yl, or pyrimidin-4-yl. In some embodiments, Cy is pyridazinyl. In some embodiments, Cy is pyrazinyl. In some embodiments, Cy is pyrimidinyl. In some embodiments, Cy is selected from:

[0141]

[0142] In some embodiments, Cy is an 8- to 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Cy is an 8- to 10-membered bicyclic heteroaryl ring having 1-3 nitrogen atoms. In some embodiments, Cy is a 10-membered bicyclic heteroaryl ring having 1-3 nitrogen atoms. In some embodiments, Cy is a 10-membered bicyclic heteroaryl ring having 1 nitrogen atom. In some such embodiments, Cy is substituted with 1 R xSubstituted. In some embodiments, Cy is quinolin-2-yl, quinolin-3-yl, quinolin-4-yl, quinolin-5-yl, quinolin-6-yl, quinolin-7-yl or quinolin-8-yl.

[0143] In some embodiments, Cy is selected from

[0144] In some embodiments, Cy is an 8- to 10-membered bicyclic aryl ring. In some embodiments, Cy is a 10-membered bicyclic aryl ring. In some such embodiments, Cy is substituted with one R x Substituted. In some embodiments, Cy is naphthalen-1-yl. In some embodiments, Cy is naphthalen-2-yl.

[0145] In some embodiments of Formula II, Cy is a 3- to 7-membered saturated or partially unsaturated carbocyclic ring, wherein Cy is substituted with 0-4 R x Substituted. In some embodiments, Cy is a 3- to 7-membered saturated carbocyclic ring. In some embodiments, Cy is cyclohexyl.

[0146] In some embodiments, R 2 and R 3 together with the nitrogen atom to which they are attached form a 4- to 7-membered saturated or partially unsaturated ring fused to Cy, or form a 4- to 7-membered saturated or partially unsaturated ring substituted with -Cy. In some embodiments, R 2 and R 3 together with the nitrogen atom to which they are attached form a ring selected from the following:

[0147]

[0148] wherein Cy is substituted with 0-4 R x Substituted.

[0149] In some embodiments, R 2 and R 3 together with the nitrogen atom to which they are attached form a 5-membered saturated ring fused to Cy. In some such embodiments, -Cy is phenyl. In some embodiments, R 2 and R 3 together with the nitrogen atom to which they are attached form

[0150] As generally defined above, each R x is independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R and optionally substituted C 1-6Aliphatic.

[0151] In some embodiments, R x is a halogen. In some such embodiments, R x is fluorine. In some embodiments, R x is chlorine. In some embodiments, R x is bromine.

[0152] In some embodiments, R x is -CN.

[0153] In some embodiments, R x is optionally substituted C 1-6 aliphatic. In some embodiments, R x is optionally substituted -C 1-6 alkyl. In some embodiments, R x is optionally halogen-substituted -C 1-6 alkyl. In some embodiments, R x is optionally substituted -CH3. In some such embodiments, R x is -CF3.

[0154] In some embodiments, R x is C 1-6 aliphatic. In some embodiments, R x is -C 1-6 alkyl. In some embodiments, R x is -CH3. In some embodiments, R x is -CH(CH3)2. In some embodiments, R x is -C(CH3)3. In some embodiments, R x is cyclopropyl.

[0155] In some embodiments, R x is -OR. In some such embodiments, R is C 1-6 aliphatic. In some embodiments, R x is -OR, where R is C 1-6 alkyl. In some embodiments, R x is -OCH3.

[0156] In some embodiments, R x is -OR. In some such embodiments, R is optionally substituted C 1-6 aliphatic. In some embodiments, R x is -OR, where R is optionally substituted C 1-6 alkyl. In some embodiments, Rx is -OR, where R is optionally substituted -CH3. In some embodiments, R x is -OR, where R is -CF3. Thus, in some embodiments, R x is -OCF3.

[0157] In some embodiments, R x is -SO2R. In some such embodiments, R is optionally substituted C 1-6 aliphatic. In some embodiments, R x is -SO2R, where R is C 1-6 alkyl. In some embodiments, R x is -SO2R, where R is -CH3. Thus, in some embodiments, R x is -SO2CH3.

[0158] In some embodiments, R x is -SO2N(R)2. In some such embodiments, R is selected from hydrogen or optionally substituted C 1-6 aliphatic. In some embodiments, R x is -SO2N(R)2, where R is selected from hydrogen and C 1-6 alkyl. In some embodiments, R x is -SO2N(R)2, where R is selected from hydrogen and -CH3. Thus, in some embodiments, R x is -SO2NHCH3 or -SO2N(CH3)2.

[0159] In some embodiments, R x is -SR. In some such embodiments, R is optionally substituted C 1-6 aliphatic. In some embodiments, R x is -SR, where R is C 1-6 alkyl. In some embodiments, R x is -SR, where R is -CH3. Thus, in some embodiments, R x is -SCH3.

[0160] In some embodiments, R x is -N(R)2. In some such embodiments, R is selected from hydrogen and optionally substituted C 1-6 aliphatic. In some embodiments, R x is -NH2, in some embodiments, R x is -NHR, where R is C 1-6 alkyl. In some embodiments, R xis -NHR, where R is -CH3. Thus, in some embodiments, R x is -NHCH3.

[0161] In some embodiments, R x is -CON(R)2. In some embodiments, R x is -CONH2.

[0162] In some embodiments, R x is -CO2R. In some embodiments, R x is -CO2H.

[0163] In some embodiments, R x is -N(R)SO2R. In some such embodiments, R is selected from hydrogen or optionally substituted C 1-6 aliphatic. In some embodiments, R x is -N(R)SO2R, where R is selected from hydrogen and C 1-6 alkyl. In some embodiments, R x is -N(R)SO2R, where R is selected from hydrogen and -CH3. Thus, in some embodiments, R x is -NHSO2CH3 or -N(CH3)SO2CH3.

[0164] As generally defined above for formula II, each R x is independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R, optionally substituted C 1-6 aliphatic group, optionally substituted 5- to 6-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur, and optionally substituted 8- to 10-membered heteroaryl ring having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur. In some embodiments of formula II, R x is an optionally substituted 5- to 6-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur. In some embodiments of formula II, R x is a 5-membered saturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur. In some such embodiments of formula II, R x is selected from

[0165] In some embodiments of formula II, R xis a 6-membered saturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some such embodiments of Formula II, R x is

[0166] In some embodiments of Formula II, R x is a 6-membered partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some such embodiments of Formula II, R x is

[0167] In some embodiments of Formula II, R x is an optionally substituted 8- to 10-membered heteroaryl ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments of Formula II, R x is an optionally substituted 9-membered heteroaryl ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some such embodiments of Formula II, R x is

[0168] In some embodiments, R 3 is selected from

[0169]

[0170]

[0171] As generally defined above, R 4 is -R. In some embodiments, R 4 is -R. In some such embodiments, -R is hydrogen. Thus, in some embodiments, R 4 is hydrogen. In some embodiments, R 4 is -R, where R is an optionally substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C 1-6 alkyl. In some embodiments, R 4 is -R, where R is C 1-3 alkyl. In some embodiments, R 4 is -R, where R is -CH3. Thus, in some embodiments, R 4 is -CH3. In some embodiments, R 4 is -R, where R is -CH2CH3. Thus, in some embodiments, R 4is -CH2CH3. In some embodiments, R 4 is -R, where R is -CH2CH2CH3. Thus, in some embodiments, R 4 is -CH2CH2CH3. In some embodiments, R 4 is -R, where R is -CH(CH3)2. Thus, in some embodiments, R 4 is -CH(CH3)2.

[0172] In some embodiments, R 4 is -R, where R is a C 1-6 aliphatic group substituted with a group selected from:-(CH2) 0-4 R o ,-(CH2) 0-4 OR o ,-(CH2) 0-4 N(R o )2,-(CH2) 0-4 S(O)2R o ,-(CH2) 0-4 C(O)R o ,-(CH2) 0-4 C(O)N(R o )2 or -(CH2) 0-4 C(O)OR o . In some embodiments, R 4 is -R, where R is a C 0-4 aliphatic group substituted with -(CH2) o R 1-6 . In some such embodiments, R o is a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R 4 is -R, where R is a C 1-6 aliphatic group substituted with a phenyl group. In some embodiments, R 4 is -R, where R is a C substituted C 1-6 aliphatic group. In some embodiments, R 4 is -R, where R is a C 1-6 aliphatic group substituted with a 5-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R 4 is -R, where R is a C substituted C 1-6 aliphatic group. In some embodiments, R 4is -R, where R is C substituted by a 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen or sulfur 1-6 aliphatic. In some embodiments, R 4 is -R, where R is substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted by an 8- to 10-membered bicyclic aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur 1-6 aliphatic. In some embodiments, R 4 is -R, where R is substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted by a 5-membered saturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur 1-6 aliphatic. In some embodiments, R 4 is -R, where R is substituted C 1-6 aliphatic.

[0173] In some embodiments, R 4 is -R, where R is C substituted by -(CH2) 0-4 OR o substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted by -OR o substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted by -OH 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted by -OCH3 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted by -OCH2CH2OCH2C≡CH 1-6 aliphatic.

[0174] In some embodiments, R 4 is -R, where R is C substituted by -(CH2) 0-4 N(R o )2 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted by -N(R o )2 1-6 aliphatic. In some embodiments, R 4is -R, where R is C substituted with -NH2 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted with -N(CH3)2 1-6 aliphatic.

[0175] In some embodiments, R 4 is -R, where R is C substituted with -(CH2) 0-4 S(O)2R o substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted with -S(O)2R o substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted with -S(O)2CH3 1-6 aliphatic.

[0176] In some embodiments, R 4 is -R, where R is C substituted with -(CH2) 0-4 C(O)N(R o )2 substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted with -C(O)N(R o )2 substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted with -C(O)NH2 1-6 aliphatic. In some embodiments, R 4 is -R, where R is C substituted with -C(O)N(R o )2 substituted C 1-6 aliphatic, where two independently occurring Rs o together with their intervening atom form a 3- to 12-membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R 4 is -R, where R is C substituted with -C(O)N(R o )2 substituted C 1-6 aliphatic, where two independently occurring Rs o together with their intervening atom form a 5- to 6-membered saturated monocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some such embodiments, R 4 is -R, where R is C substituted with substituted C 1-6 aliphatic. In some embodiments, R 4is -R, where R is a substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is a substituted C 1-6 aliphatic.

[0177] In some embodiments, R 4 is -R, where R is a C substituted by -(CH2) 0-4 C(O)OR o substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is a C substituted by -C(O)OR o substituted C 1-6 aliphatic. In some embodiments, R 4 is -R, where R is a C substituted by -C(O)OH 1-6 aliphatic.

[0178] In some embodiments, R 4 is H, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2 or a group selected from:

[0179]

[0180] As generally defined above, each R is independently hydrogen or an optionally substituted C 1-6 aliphatic; or two instances of R together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocycle.

[0181] In some embodiments, R is hydrogen. In some embodiments, R is an optionally substituted C 1-6 aliphatic. In some such embodiments, R is -C 1-6 alkyl. In some embodiments, R is -CH3. In some embodiments, R is an optionally halogen-substituted C 1-6 aliphatic. In some embodiments, R is an optionally C substituted by -(CH2) 0-4 R o substituted C 1-6 aliphatic. In some embodiments, R is an optionally C substituted by -R o substituted C 1-6 aliphatic. In some embodiments, R is an optionally C substituted by -(CH2) 0-4 OR o substituted C 1-6 aliphatic. In some embodiments, R is an optionally C substituted by -OR o substituted C 1-6Aliphatic. In some embodiments, R is optionally substituted with -(CH2) 0-4 N(R o )2-substituted C 1-6 Aliphatic. In some embodiments, R is optionally substituted with -N(R o )2-substituted C 1-6 Aliphatic. In some embodiments, R is optionally substituted with -(CH2) 0-4 C(O)OR o -substituted C 1-6 Aliphatic. In some embodiments, R is optionally substituted with -C(O)OR o -substituted C 1-6 Aliphatic. In some embodiments, R is optionally substituted with -(CH2) 0-4 C(O)N(R o )2-substituted C 1-6 Aliphatic. In some embodiments, R is optionally substituted with -C(O)N(R o )2-substituted C 1-6 Aliphatic. In some embodiments, R is optionally substituted with -(CH2) 0-4 S(O)2R o -substituted C 1-6 Aliphatic. In some embodiments, R is optionally substituted with -S(O)2R o -substituted C 1-6 Aliphatic.

[0182] In some embodiments, two instances of R together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocycle. In some embodiments, two instances of R together with the atoms to which they are attached form a 6-membered saturated or partially unsaturated heterocycle. In some embodiments, two instances of R together with the atoms to which they are attached form a pyrrolidinyl ring. In some embodiments, two instances of R together with the atoms to which they are attached form a morpholinyl ring. In some embodiments, two instances of R together with the atoms to which they are attached form a piperidinyl ring. In some embodiments, two instances of R together with the atoms to which they are attached form a 2-azabicyclo[2.2.2]octyl ring.

[0183] As defined generally above for formula II, each R is independently hydrogen, optionally substituted C 1-6 aliphatic, optionally substituted phenyl, or optionally substituted 3- to 7-membered saturated or partially unsaturated carbocycle, or two instances of R together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocycle. In some embodiments of formula II, R is hydrogen. In some embodiments of formula II, R is optionally substituted C 1-6Aliphatic. In some such embodiments, R is -C 1-6 alkyl. In some embodiments of Formula II, R is -CH3.

[0184] In some embodiments of Formula II, R is C optionally substituted with halogen 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -(CH2) 0-4 R o substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -R o substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -(CH2) 0-4 OR o substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -OR o substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -(CH2) 0-4 N(R o )2 substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -N(R o )2 substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -(CH2) 0-4 C(O)OR o substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -C(O)OR o substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -(CH2) 0-4 C(O)N(R o )2 substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -C(O)N(R o )2 substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -(CH2) 0-4 S(O)2R o substituted C 1-6 aliphatic. In some embodiments of Formula II, R is optionally substituted with -S(O)2R o substituted C 1-6 aliphatic.

[0185] In some embodiments of Formula II, two instances of R together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocycle. In some embodiments of Formula II, two instances of R together with the atoms to which they are attached form a 6-membered saturated or partially unsaturated heterocycle. In some embodiments of Formula II, two instances of R together with the atoms to which they are attached form a pyrrolidinyl ring. In some embodiments of Formula II, two instances of R together with the atoms to which they are attached form a morpholinyl ring. In some embodiments of Formula II, two instances of R together with the atoms to which they are attached form a piperidinyl ring. In some embodiments of Formula II, two instances of R together with the atoms to which they are attached form a 2-azabicyclo[2.2.2]octyl ring.

[0186] It is to be understood that a compound of Formula I or Formula II having the following structure

[0187]

[0188] When R 4 is H, it can exist in two tautomeric forms:

[0189]

[0190] Thus, it is to be understood that when R 4 is H, the compound of Formula I or Formula II can be drawn in either tautomeric form.

[0191] In some embodiments of Formula I or Formula II, R 1 is -CN. Thus, in some embodiments, the present disclosure provides a compound of Formula I-a:

[0192]

[0193] or a pharmaceutically acceptable salt thereof, wherein each of R 2 , R 3 and R 4 is as defined above and described herein.

[0194] In some embodiments of Formula I or Formula II, R 1 is -CON(R)2. Thus, in some embodiments, the present disclosure provides a compound of Formula I-b:

[0195]

[0196] or a pharmaceutically acceptable salt thereof, wherein each of R 2 , R 3 , R 4 and R is as defined above and described herein.

[0197] In some embodiments of Formula I or Formula II, R 1 is -C(=NR)N(R)2. Thus, in some embodiments, the present disclosure provides a compound of Formula I-c:

[0198]

[0199] or a pharmaceutically acceptable salt thereof, wherein each of R 2 , R 3 , R 4 and R is as defined above and described herein.

[0200] In some embodiments of Formula I-a, I-b or I-c, R 2 is H. Thus, the present disclosure provides a compound of Formula I-a-i, I-b-i or I-c-i:

[0201]

[0202] or a pharmaceutically acceptable salt thereof, wherein each of R 3 , R 4 and R is as defined above and described herein.

[0203] In some embodiments of Formula I-a-i, I-b-i or I-c-i, R 3 is Cy. Thus, the present disclosure provides a compound of Formula I-a-ii, I-b-ii or I-c-ii:

[0204]

[0205] or a pharmaceutically acceptable salt thereof, wherein each of Cy, R 4 and R is as defined above and described herein.

[0206] In some embodiments of Formula I-a, I-b or I-c, R 4 is hydrogen. Thus, the present disclosure provides a compound of Formula I-a-iii, I-b-iii or I-c-iii:

[0207]

[0208] or a pharmaceutically acceptable salt thereof, wherein each of R 2 , R 3 and R is as defined above and described herein.

[0209] In some embodiments of Formula I-a-iii, I-b-iii or I-c-iii, R 3It is Cy. Accordingly, the present disclosure provides a compound of formula I-a-iv, I-b-iv or I-c-iv:

[0210]

[0211] or a pharmaceutically acceptable salt thereof, wherein each of Cy, R 2 and R is as defined above and described herein.

[0212] In some embodiments of formula I-a, I-b or I-c, R 3 is -Cy, wherein -Cy is a phenyl group substituted with 0-4 R x groups. Accordingly, in some embodiments, the present disclosure provides a compound of formula I-a-v, I-b-v or I-c-v:

[0213]

[0214]

[0215] or a pharmaceutically acceptable salt thereof, wherein R 2 、R 4 、R and R x each is as defined above and described herein.

[0216] In some embodiments of formula I-a, I-b or I-c, R 3 is -Cy, wherein -Cy is a quinolin-5-yl group substituted with 0-4 R x groups. Accordingly, in some embodiments, the present disclosure provides a compound of formula I-a-vi, I-b-vi or I-c-vi:

[0217]

[0218] or a pharmaceutically acceptable salt thereof, wherein R 2 、R 4 、R and R x each is as defined above and described herein.

[0219] In some embodiments, the present disclosure provides a compound selected from:

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237] Tool compound

[0238] In some embodiments, one or more compounds of Formula I or Formula II are tethered to a detectable moiety to form a tool compound. In some embodiments, the tool compound comprises a compound of Formula I or Formula II, a detectable moiety, and a tether moiety that links the detectable moiety to the compound of Formula I or Formula II. In some embodiments, the tool compound comprises a compound of Formula I or Formula II and a moiety comprising a functional group capable of binding or reacting with the detectable moiety.

[0239] In some embodiments, the present disclosure provides a compound of Formula III:

[0240]

[0241] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 and R 3 are each as defined above for Formula II and are described in each collection and subset herein; T is a divalent tether moiety; and R t is a detectable moiety.

[0242] In some embodiments, R tis a detectable moiety selected from a primary label or a secondary label. In certain embodiments, R t is a detectable moiety selected from a fluorescent label (e.g., a fluorescent dye or a chromophore), a mass tag, a chemiluminescent group, a chromophore, an electron-dense group, and an energy transfer agent.

[0243] As used herein, the term "detectable moiety" is used interchangeably with the terms "label" and "reporter molecule" and refers to any moiety that can be detected, such as a primary label and a secondary label. The presence of a detectable moiety can be measured by using methods that quantify (in an absolute, approximate, or relative manner) the detectable moiety in the system under study. In some embodiments, such methods are well known to those of ordinary skill in the art and include any method of quantifying a reporter molecule moiety (e.g., a label, a dye, a photo-crosslinker, a cytotoxic compound, a drug, an affinity label, a photoaffinity label, a reactive compound, an antibody or antibody fragment, a biomaterial, a nanoparticle, a spin label, a chromophore, a metal-containing moiety, a radioactive moiety, a quantum dot, a novel functional group, a group that covalently or non-covalently interacts with other molecules, a photocaged moiety, a photoactivatable radiation-excitable moiety, a ligand, a photoisomerizable moiety, biotin, a biotin analog (e.g., biotin sulfoxide), a moiety incorporating a heavy atom, a chemically cleavable group, a photocleavable group, a redox-active agent, an isotope-labeled moiety, a biophysical probe, a phosphorescent group, a chemiluminescent group, an electron-dense group, a magnetic group, an intercalating group, a chromophore, an energy transfer agent, a bioactive agent, a detectable label, and any combination of the foregoing).

[0244] Primary labels such as radioactive isotopes (e.g., tritium, 32 P, 33 P, 35 S, 14 C, 123 I, 124 I, 125 I or 131 I), mass tags including but not limited to stable isotopes (e.g., 13 C, 2 H, 17 O, 18 O, 15 N, 19 F and 127 I), positron emitting isotopes (e.g., 11 C, 18 F, 13 N, 124 I and 15O) and fluorescent labels, are signal generating reporter groups that can be detected without further modification. The detectable moiety can be analyzed by methods including but not limited to fluorescence, positron emission tomography, SPECT medical imaging, chemiluminescence, electron spin resonance, ultraviolet / visible absorbance spectroscopy, mass spectrometry, nuclear magnetic resonance, magnetic resonance, flow cytometry, autoradiography, scintillation counting, phosphorescence imaging, and electrochemical methods.

[0245] As used herein, the term "secondary label" refers to moieties such as biotin and various protein antigens that require the presence of a second intermediate to generate a detectable signal. For biotin, the second intermediate can include a streptavidin-enzyme conjugate. For antigen labeling, the second intermediate can include an antibody-enzyme conjugate. Some fluorophores act as secondary labels because they transfer energy to another moiety during a non-radiative fluorescence resonance energy transfer (FRET) process, and this second moiety generates the detection signal.

[0246] As used herein, the terms "fluorescent label", "fluorescent dye", and "chromophore" refer to moieties that absorb light energy at a defined excitation wavelength and emit light energy at a different wavelength. Examples of fluorescent labels include, but are not limited to: Alexa Fluor dyes (Alexa Fluor 350, Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 660, and Alexa Fluor 680), AMCA, AMCA-S, BODIPY dyes (BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 493 / 503, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, BODIPY 650 / 665), carboxyrhodamine 6G, carboxy-X-rhodamine (ROX), cascade blue, cascade yellow, coumarin 343, cyanine dyes (Cy3, Cy5, Cy3.5, Cy5.5), dansyl, Dapoxyl, dialkylaminocoumarin, 4',5'-dichloro-2',7'-dimethoxy-fluorescein, DM-NERF, eosin, erythrosin, fluorescein, FAM, hydroxycoumarin, IRDye (IRD40, IRD 700, IRD 800), JOE, Lissamine rhodamine B, MarinaBlue, methoxycoumarin, naphthofluorescein, Oregon Green 488, Oregon Green 500, Oregon Green 514, Pacific Blue, PyMPO, pyrene, rhodamine B, rhodamine 6G, rhodamine green, rhodamine red, Rhodol Green, 2',4',5',7'-tetrabromosulfone-fluorescein, tetramethyl-rhodamine (TMR), carboxytetramethylrhodamine (TAMRA), Texas Red, Texas Red-X, 5(6)-carboxyfluorescein, 2,7-dichlorofluorescein, N,N-bis(2,4,6-trimethylphenyl)-3,4:9,10 - Perylene bis(dicarboximide), HPTS, Ethyl Eosin, DY - 490XL MegaStokes, DY - 485XL MegaStokes, Adirondack Green 520, ATTO465, ATTO 488, ATTO 495, YOYO - 1, 5 - FAM, BCECF, Dichlorofluorescein, Rhodamine 110, Rhodamine 123, YO - PRO - 1, SYTOX Green, Sodium Green, SYBR Green I, Alexa Fluor 500, FITC, Fluo - 3, Fluo - 4, fluoro - emerald, YoYo - 1 ssDNA, YoYo - 1 dsDNA, YoYo - 1, SYTO RNASelect, Diversa Green - FP, Dragon Green, EvaGreen, Surf Green EX, Spectrum Green, NeuroTrace 500 / 525, NBD - X, MitoTracker Green FM, LysoTracker Green DND - 26, CBQCA, PA - GFP (after activation), WEGFP (after activation), FlASH - CCXXCC, Azami Green monomeric, Azami Green, Green Fluorescent Protein (GFP), EGFP (Campbell Tsien 2003), EGFP (Patterson 2001), Kaede Green, 7 - Benzylamino - 4 - nitrobenz - 2 - oxa - 1,3 - dioxole, Bexl, Doxorubicin, LumioGreen, and SuperGlo GFP.,

[0247] As used herein, the term "mass tag" refers to any moiety that can be uniquely detected by virtue of its mass using mass spectrometry (MS) detection techniques. Examples of mass tags include electrophoretically released tags such as N-[3-[4'-[(p-methoxytetrafluorobenzyl)oxy]phenyl]-3-methylglyceryl]isopiperidinecarboxylic acid, 4'-[2,3,5,6-tetrafluoro-4-(pentafluorophenoxy)]acetophenone, and derivatives thereof. The synthesis and utility of these mass tags are described in U.S. Patents 4,650,750, 4,709,016, 5,360,8191, 5,516,931, 5,602,273, 5,604,104, 5,610,020, and 5,650,270. Other examples of mass tags include, but are not limited to, nucleotides, dideoxynucleotides, oligonucleotides of different lengths and base compositions, oligopeptides, oligosaccharides, and other synthetic polymers of different lengths and monomer compositions. A variety of neutral and charged organic molecules (biomolecules or synthetic compounds) within an appropriate mass range (100 - 2000 daltons) can also be used as mass tags. Stable isotopes (e.g., 13 C, 2 H, 17 O, 18 O, and 15 N) can also be used as mass tags.

[0248] As used herein, the term "chemiluminescent group" refers to a group that emits light as a result of a chemical reaction without the addition of heat. For example, luminol (5-amino-2,3-dihydro-1,4-phthalhydrazide) reacts with an oxidizing agent such as hydrogen peroxide (H2O2) in the presence of a base and a metal catalyst to produce an excited-state product (3-aminophthalic acid, 3-APA).

[0249] As used herein, the term "chromophore" refers to a molecule that absorbs light in the visible, UV, or IR wavelength range.

[0250] As used herein, the term "dye" refers to a soluble coloring substance that contains a chromophore.

[0251] As used herein, the term "electron-dense group" refers to a group that scatters electrons when irradiated with an electron beam. Such groups include, but are not limited to, ammonium molybdate, bismuth subnitrate, cadmium iodide, carbohydrazide, ferric chloride hexahydrate, hexamethylenetetramine, indium trichloride anhydrous, lanthanum nitrate, lead acetate trihydrate, lead citrate trihydrate, lead nitrate, periodic acid, phosphomolybdic acid, phosphotungstic acid, potassium ferricyanide, potassium ferrocyanide, ruthenium red, silver nitrate, silver proteinate (Ag assay: 8.0 - 8.5%) "Strong", silver tetraphenylporphine (S-TPPS), sodium chloroaurate, sodium tungstate, thallium nitrate, thiosemicarbazide (TSC), uranyl acetate, uranyl nitrate, and vanadyl sulfate.

[0252] As used herein, the term "energy transfer agent" refers to a molecule that supplies energy to another molecule or accepts energy from another molecule. By way of example only, fluorescence resonance energy transfer (FRET) is a dipole-dipole coupling process by which the excited state energy of a fluorescent donor molecule is transferred non-radiatively to an unexcited acceptor molecule, which then emits the supplied energy as fluorescence at a longer wavelength.

[0253] As used herein, the term "moiety incorporating a heavy atom" refers to a group incorporating an ion of an atom that is generally heavier than carbon. In some embodiments, such ions or atoms include, but are not limited to, silicon, tungsten, gold, lead, and uranium.

[0254] As used herein, the term "photoaffinity label" refers to a label bearing a group that forms a linkage with a molecule having an affinity for the label upon exposure to light.

[0255] As used herein, the term "photocaging moiety" refers to a group that binds covalently or non-covalently to other ions or molecules upon irradiation at certain wavelengths.

[0256] As used herein, the term "photoisomerizable moiety" refers to a group that changes from one isomeric form to another when irradiated with light.

[0257] As used herein, the term "radioactive moiety" refers to a group whose atomic nucleus spontaneously emits nuclear radiation (such as α, β, or γ particles); where an α particle is a helium nucleus, a β particle is an electron, and a γ particle is a high-energy photon.

[0258] As used herein, the term "spin label" refers to a molecule containing an atom or group of atoms (i.e., a stable paramagnetic group) that exhibits an unpaired electron spin, which in some embodiments is detected by electron spin resonance spectroscopy and in other embodiments is attached to another molecule. Such spin-label molecules include, but are not limited to, nitroxide radicals and nitroxy groups, and in some embodiments are single spin labels or double spin labels.

[0259] As used herein, the term "quantum dot" refers to colloidal semiconductor nanocrystals that, in some embodiments, are detected in the near infrared and have an extremely high quantum yield (i.e., are very bright under moderate illumination).

[0260] One of ordinary skill in the art will recognize that the detectable moiety can be attached to the provided compounds by suitable substituents. As used herein, the term "suitable substituent" refers to a moiety capable of covalently linking to the detectable moiety. Such moieties are well known to those of ordinary skill in the art and include groups containing, for example, carboxylate moieties, amino moieties, thiol moieties, or hydroxyl moieties (to name just a few). It is understood that such moieties can be attached directly or through a tether moiety, such as a divalent saturated or unsaturated hydrocarbon chain, to the provided compounds.

[0261] In some embodiments, -T- is selected from -(CH2CH2O) n -, -(C 1-6 alkyl)N(R)C(O)(C 1-6 alkyl)-, and -(CH2CH2O) n (C 1-6 alkyl)N(R)C(O)(C 1-6 alkyl)-, where n is 1 - 4. In some embodiments, -T- is selected from -(CH2CH2O) n -, -(C 3-5 alkyl)N(R)C(O)(C 2-4 alkyl)-, and -(CH2CH2O) n (C 3-5 alkyl)N(R)C(O)(C 2-4 alkyl)-, where n is 2 - 3. In some embodiments, -T- is selected from the group consisting of:

[0262]

[0263] In some embodiments, the detectable moiety R t is attached to the compound of Formula I or Formula II by click chemistry. In some embodiments, the compound of Formula I or Formula II is attached to -T-R t by a 1,3-cycloaddition of an azide and an alkyne (optionally in the presence of a copper catalyst). Methods using click chemistry are known in the art and include those described by Rostovtsev et al., Angew. Chem. Int. Ed. 2002, 41 , 2596 - 99 and Sun et al., Bioconjugate Chem., 2006, 17 , 52 - 57. In some embodiments, the compound of Formula IV is a click ready inhibitor. In some such embodiments, the click ready inhibitor of Formula IV reacts with the click ready -T-Rt Partial reaction. As used herein, "clickable moiety" refers to a moiety containing an azide or an alkyne for click chemistry reactions. In some embodiments, the clickable inhibitor moiety contains an azide. In certain embodiments, the clickable-T-R t moiety contains a strained cyclooctyne for copper-free click chemistry reactions (e.g., using the method described in Baskin et al., Proc. Natl. Acad. Sci. USA 2007, 104 , 16793 - 16797).

[0264] In some embodiments, the compounds of Formula III are selected from

[0265]

[0266] In some embodiments, one or more compounds of Formula I or Formula II covalently inhibit SARM1. In some embodiments, one or more compounds of Formula I or Formula II covalently modify a cysteine residue of SARM1. In some embodiments, one or more compounds of Formula I or Formula II covalently modify Cys635 of SARM1. In some embodiments, one or more compounds of Formula I or Formula II covalently modify Cys629 of SARM1. In some embodiments, one or more compounds of Formula I or Formula II covalently modify Cys649 of SARM1. Without wishing to be bound by any particular theory, in some embodiments, one or more compounds of Formula I or Formula II denature the SARM1 protein through covalent modification of a cysteine residue. In some embodiments, one or more compounds of Formula I or Formula II denature the SARM1 protein through covalent modification of Cys635. In some embodiments, one or more compounds of Formula I or Formula II denature the SARM1 protein through covalent modification of Cys629. In some embodiments, one or more compounds of Formula I or Formula II denature the SARM1 protein through covalent modification of Cys649.

[0267] In some aspects, the present disclosure provides compounds according to the following embodiments:

[0268] Embodiment 1. A compound having Formula II:

[0269]

[0270] or a pharmaceutically acceptable salt thereof, wherein

[0271] R 1Selected from -CN, -NO2, -C(O)R, -S(O)2R, -CON(R)2, -S(O)2N(R)2, -CO2R, -C(=NR)N(R)2

[0272] and a 5-membered heteroaryl ring having 2-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur;

[0273] R 2 is -R;

[0274] R 3 is -(CH2) 0-2 Cy, or:

[0275] R 2 and R 3 together with the nitrogen atom to which they are attached form a 4- to 7-membered saturated or partially unsaturated ring fused to Cy, or form a 4- to 7-membered saturated or partially unsaturated ring substituted with -Cy;

[0276] Cy is selected from phenyl, a 3- to 7-membered saturated or partially unsaturated carbocyclic ring, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, an 8- to 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8- to 10-membered bicyclic aryl ring, wherein each phenyl, carbocyclic ring, heteroaryl ring, and aryl ring is substituted with 0-4 R x substituents;

[0277] Each R x is independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R, optionally substituted C 1-6 aliphatic group, an optionally substituted 5- to 6-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an optionally substituted 8- to 10-membered heteroaryl ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur;

[0278] R 4 is -R; and

[0279] each R is independently hydrogen, optionally substituted C 1-6 aliphatic, optionally substituted phenyl, and optionally substituted 3- to 7-membered saturated or partially unsaturated carbocyclic ring, or:

[0280] Examples of two Rs together with the atom to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocycle.

[0281] Embodiment 2. A compound having the formula I:

[0282]

[0283] or a pharmaceutically acceptable salt thereof, wherein

[0284] R 1 is selected from -CN, -NO2, -C(O)R, -S(O)2R, -CON(R)2, -S(O)2N(R)2, and -CO2R;

[0285] R 2 is -R;

[0286] R 3 is selected from -(CH2) 0-2 Cy, or:

[0287] R 2 and R 3 together with the nitrogen atom to which they are attached form a 4- to 7-membered saturated or partially unsaturated ring fused to Cy, or form a 4- to 7-membered saturated or partially unsaturated ring substituted with -Cy;

[0288] Cy is selected from phenyl, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, an 8- to 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8- to 10-membered bicyclic aryl ring, wherein each phenyl, heteroaryl ring, and aryl ring is substituted with 0-4 R x substituents;

[0289] Each R x is independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R, and optionally substituted C 1-6 aliphatic;

[0290] R 4 is -R;

[0291] Each R is independently hydrogen or optionally substituted C 1-6 aliphatic; or:

[0292] Examples of two Rs together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocycle.

[0293] Embodiment 3. The compound according to Embodiment 1 or Embodiment 2, wherein R 2 is -H.

[0294] Embodiment 4. The compound according to Embodiment 1 or Embodiment 2, wherein R 2is -R.

[0295] Embodiment 5. The compound according to Embodiment 4, wherein -R is C 1-6 aliphatic.

[0296] Embodiment 6. The compound according to Embodiment 5, wherein R is -C 1-6 alkyl.

[0297] Embodiment 7. The compound according to Embodiment 6, wherein R is -CH3.

[0298] Embodiment 8. The compound according to any one of Embodiments 1-7, wherein R 1 is -CN.

[0299] Embodiment 9. The compound according to any one of Embodiments 1-7, wherein R 1 is -C(O)N(R)2.

[0300] Embodiment 10. The compound according to any one of Embodiments 1-7, wherein R 1 is selected from

[0301]

[0302] Embodiment 11. The compound according to any one of Embodiments 1-7, wherein R 1 is -CO2R.

[0303] Embodiment 12. The compound according to any one of Embodiments 1-7, wherein R 1 is -NO2.

[0304] Embodiment 13. The compound according to any one of Embodiments 1-7, wherein R 1 is -C(O)R.

[0305] Embodiment 14. The compound according to any one of Embodiments 1-7, wherein R 1 is -S(O)2R.

[0306] Embodiment 15. The compound according to any one of Embodiments 1-7, wherein R 1 is -S(O)2N(R)2.

[0307] Embodiment 16. The compound according to any one of Embodiments 1-7, wherein R 1 is -C(=NR)C(R)2.

[0308] Embodiment 17. The compound according to Embodiment 16, wherein R 1 is selected from

[0309]

[0310] Embodiment 18. The compound according to any one of Embodiments 1-7, wherein R 1 is a 5-membered heteroaryl ring having 2-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0311] Embodiment 19. The compound according to Embodiment 18, wherein R 1 is

[0312] Embodiment 20. The compound according to any one of Embodiments 1-19, wherein R 4 is -H.

[0313] Embodiment 21. The compound according to any one of Embodiments 1-19, wherein R 4 is -R, where -R is an optionally substituted C 1-6 aliphatic.

[0314] Embodiment 22. The compound according to Embodiment 21, wherein R is -C 1-6 alkyl.

[0315] Embodiment 23. The compound according to Embodiment 22, wherein R is -CH3.

[0316] Embodiment 24. The compound according to Embodiment 22, wherein R is -CH2CH3.

[0317] Embodiment 25. The compound according to Embodiment 22, wherein R is -CH2CH2CH3.

[0318] Embodiment 26. The compound according to Embodiment 22, wherein R is -CH(CH3)2.

[0319] Embodiment 27. The compound according to Embodiment 21, wherein R is selected from -CH2C(O)NH2, -CH2CH2N(CH3)2, -CH2CH2OH, -CH2CH(CH3)OH, -CH(CH3)CH2OH, -CH2C(CH3)2OH, -CH2CH2OCH3, -CH2C(CH3)2OCH3, -CH(CH3)CH2OCH3, -CH2CH2C(O)NH2, -CH2CH2C(O)OH, -CH2CH2S(O)2CH3, -CH2CH2CH2OCH3,

[0320] Embodiment 28. The compound according to any one of Embodiments 1-27, wherein R 3 is -Cy.

[0321] Embodiment 29. The compound according to any one of Embodiments 1-27, wherein R 3 is -CH2Cy.

[0322] Embodiment 30. The compound according to Embodiment 28 or Embodiment 29, wherein -Cy is phenyl.

[0323] Embodiment 31. The compound according to Embodiment 28 or Embodiment 29, wherein -Cy is a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0324] Embodiment 32. The compound according to Embodiment 31, wherein -Cy is a 6-membered heteroaryl ring having 1-3 nitrogen atoms.

[0325] Embodiment 33. The compound according to Embodiment 32, wherein -Cy is pyridyl.

[0326] Embodiment 34. The compound according to Embodiment 28 or Embodiment 29, wherein -Cy is an 8- to 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0327] Embodiment 35. The compound according to Embodiment 34, wherein -Cy is a 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0328] Embodiment 36. The compound according to Embodiment 35, wherein -Cy is a 10-membered bicyclic heteroaryl ring having 1 nitrogen atom.

[0329] Embodiment 37. The compound according to Embodiment 36, wherein -Cy is quinolinyl.

[0330] Embodiment 38. The compound according to Embodiment 28 or Embodiment 29, wherein -Cy is an 8- to 10-membered bicyclic aryl ring.

[0331] Embodiment 39. The compound according to Embodiment 38, wherein -Cy is a 10-membered bicyclic aryl ring.

[0332] Embodiment 40. The compound according to Embodiment 39, wherein -Cy is 1-naphthyl.

[0333] Embodiment 41. The compound according to Embodiment 39, wherein -Cy is 2-naphthyl.

[0334] Embodiment 42. The compound according to Embodiment 28, wherein -Cy is selected from the group consisting of:

[0335]

[0336]

[0337] Embodiment 43. The compound according to Embodiment 28, wherein -Cy is selected from the group consisting of:

[0338]

[0339] Embodiment 44. The compound according to any one of Embodiments 28-43, wherein R x is a halogen.

[0340] Embodiment 45. The compound according to Embodiment 44, wherein R x is chlorine.

[0341] Embodiment 46. The compound according to Embodiment 44, wherein R x is fluorine.

[0342] Embodiment 47. The compound according to any one of Embodiments 28-43, wherein R x is optionally substituted -C 1-6 aliphatic.

[0343] Embodiment 48. The compound according to Embodiment 47, wherein R x is -C 1-6 alkyl.

[0344] Embodiment 49. The compound according to Embodiment 48, wherein R x is -CH3.

[0345] Embodiment 50. The compound according to Embodiment 48, wherein R x is -CH(CH3)2.

[0346] Embodiment 51. The compound according to Embodiment 47, wherein R x is optionally halogen-substituted -C 1-6 aliphatic.

[0347] Embodiment 52. The compound according to Embodiment 51, wherein R x is -CF3.

[0348] Embodiment 53. The compound according to any one of Embodiments 28-43, wherein R x is -OR.

[0349] Embodiment 54. The compound according to embodiment 53, wherein R is optionally substituted -C 1-6 aliphatic.

[0350] Embodiment 55. The compound according to embodiment 54, wherein R is -C 1-6 alkyl.

[0351] Embodiment 56. The compound according to embodiment 55, wherein R is -CH3.

[0352] Embodiment 57. The compound according to embodiment 54, wherein R is optionally halogen-substituted -C 1-6 aliphatic.

[0353] Embodiment 58. The compound according to embodiment 57, wherein R is -CF3.

[0354] Embodiment 59. The compound according to any one of embodiments 28-43, wherein R x is -SR.

[0355] Embodiment 60. The compound according to embodiment 59, wherein -R is optionally substituted -C 1-6 aliphatic.

[0356] Embodiment 61. The compound according to embodiment 60, wherein -R is -C 1-6 alkyl.

[0357] Embodiment 62. The compound according to embodiment 61, wherein -R is -CH3.

[0358] Embodiment 63. The compound according to any one of embodiments 28-43, wherein -R x is -SO2R.

[0359] Embodiment 64. The compound according to embodiment 63, wherein R is -C 1-6 alkyl.

[0360] Embodiment 65. The compound according to embodiment 64, wherein R is -CH3.

[0361] Embodiment 66. The compound according to embodiment 1 or embodiment 2, wherein:

[0362] R 1 is selected from -CN and -CONH2; and

[0363] R 2 is selected from -H and -CH3.

[0364] Embodiment 67. The compound according to Embodiment 1 or Embodiment 2, wherein:

[0365] R 1 is selected from -CN and -CONH2;

[0366] R 2 is selected from -H and -CH3;

[0367] R 3 is -Cy; and

[0368] -Cy is selected from

[0369]

[0370] Embodiment 68. The compound according to Embodiment 67, wherein:

[0371] R 1 is -CN; and

[0372] R 2 is -H.

[0373] Embodiment 69. The compound according to Embodiment 67, wherein:

[0374] R 1 is -CONH2; and

[0375] R 2 is -H.

[0376] Preparation of a compound of formula I or formula II

[0377] The compound represented by Formula I or Formula II can be prepared according to the following Schemes Ia and Ib.

[0378] Scheme Ia:

[0379]

[0380] In Scheme Ia, thiophosgene A and amine B are coupled together in the presence of a suitable base to obtain isocyanate C. In the presence of a suitable base, isocyanate C is treated with amide D to obtain thioamide E, which is treated with bromine to obtain the compound represented by General Formula I or General Formula II, wherein R 2 is H and R 1 is -CN or -CONH2, the latter being produced by hydrolysis of CN in this step.

[0381] Scheme Ib:

[0382]

[0383] In Scheme Ib, compound F is treated with a suitable base and compound G, where X is a suitable leaving group, to give a compound of formula I or formula II, where R 2 is -C 1-6 alkyl.

[0384] Composition

[0385] In some embodiments, the compounds of formula I or formula II can be provided, for example, in the form of a composition that is combined (e.g., mixed) with one or more other components.

[0386] In some embodiments, the present disclosure provides a composition that comprises and / or delivers a compound of formula I or formula II or an active metabolite thereof, e.g., when contacted with or otherwise administered to a system or environment, such as a system or environment that can include SARM1 NAD enzyme activity; in some embodiments, administering the composition to the system or environment can achieve inhibition of SARM1 activity as described herein.

[0387] In some embodiments, the provided composition herein can be a pharmaceutical composition, where the pharmaceutical composition comprises an active agent and one or more pharmaceutically acceptable excipients; in some such embodiments, the provided pharmaceutical composition comprises a compound of formula I or formula II or an active metabolite thereof and / or delivers a compound of formula I or formula II or an active metabolite thereof to a relevant system or environment as described herein (e.g., to a subject in need).

[0388] In some embodiments, one or more compounds of formula I or formula II are provided and / or utilized in the form of a pharmaceutically acceptable salt.

[0389] Among other things, the present disclosure provides a composition comprising a compound of Formula I or Formula II, or a pharmaceutically acceptable salt or derivative thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of the compound in the provided composition is such that axonal degeneration in a biological sample or patient is effectively and measurably inhibited. In certain embodiments, the provided compound or composition is formulated for administration to a patient in need of such a composition. According to the methods of the present disclosure, the compounds and compositions may be administered in any amount and by any route of administration effective to treat or alleviate the severity of any disease or disorder described herein. The provided compounds are preferably formulated in unit dosage forms for ease of administration and uniformity of dosage. As used herein, the expression "unit dosage form" refers to physically discrete units of medicament suitable for the patient to be treated. However, it is to be understood that the total daily usage of the provided compounds and compositions will be decided by the attending physician within the scope of sound medical judgment. The specific effective dosage level for any particular patient or organism will vary depending on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed and its route of administration; the species, age, weight, sex, and diet of the patient; the general condition of the subject; the time of administration; the excretion rate of the specific compound employed; the duration of the treatment; drugs used in combination with or concurrently with the specific compound employed, and the like.

[0390] The provided composition may be administered orally, parenterally, by inhalation or nasal spray, topically (e.g., by powder, ointment, or drops), rectally, buccally, vaginally, intraperitoneally, intracisternally, or by an implantable reservoir, depending on the severity of the condition being treated. Preferably, the composition is administered orally, intraperitoneally, or intravenously. In certain embodiments, the provided compound is administered orally or parenterally at a dosage level of about 0.01 mg / kg to about 50 mg / kg of the subject's body weight, one or more times a day, to achieve the desired therapeutic effect.

[0391] As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intraliver, intralesional, and intracranial injection or infusion techniques. The sterile injectable form of the provided composition may be an aqueous or oily suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, a sterile, fixed oil is conventionally employed as a solvent or suspending medium.

[0392] For this purpose, any mild non-volatile oil can be employed, including synthetic monoglycerides or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives can be used for preparing injectables, and natural pharmaceutically acceptable oils such as olive oil or castor oil, especially their polyoxyethylated forms, can also be used for preparing injectables. These oil solutions or suspensions can also contain long-chain alcohol diluents or dispersants such as carboxymethyl cellulose or similar dispersants which are commonly used in formulating pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants such as Tweens, Spans and other emulsifiers or bioavailability enhancers which are commonly used in manufacturing pharmaceutically acceptable solid, liquid or other dosage forms can also be used for formulating purposes.

[0393] Injectable preparations can be sterilized, for example, by filtration through a bacteria-retaining filter or incorporated with 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 before use.

[0394] To prolong the action of the provided compounds, it is generally desirable to slow down the absorption of the compounds injected subcutaneously or intramuscularly. This can be achieved by using a liquid suspension of a poorly water-soluble crystalline or amorphous material. Then, the absorption rate of the compound depends on its dissolution rate which in turn can depend on the crystal size and crystalline form. Alternatively, delayed absorption of the parenterally administered compound form is achieved by dissolving or suspending the compound in an oily vehicle. Injectable depot forms are made by forming a microcapsule matrix of the compound in a biodegradable polymer such as poly(lactide-co-glycolide). The release rate of the compound can be controlled according to the ratio of the compound to the polymer and the nature of the particular polymer employed. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable preparations can also be prepared by encapsulating the compound in liposomes or microemulsions compatible with body tissues.

[0395] The pharmaceutically acceptable compositions described herein can be orally administered 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 admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also conventionally contain additional substances in addition to the inert diluents such as lubricating agents and other tabletting aids such as magnesium stearate and microcrystalline cellulose. When an aqueous suspension for oral use is required, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents can also be added.

[0396] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dibasic calcium phosphate, and / or a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; c) humectants such as glycerol; d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) solution retarders such as paraffin; f) absorption promoters such as quaternary ammonium compounds; g) wetting agents such as cetyl alcohol and glycerol monostearate; h) absorbents such as kaolin and bentonite; and / or i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents. The active compounds can also be in microencapsulated form with one or more of the excipients described above.

[0397] Solid compositions of a similar type can also be used as fillers in soft and hard gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols. Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings (i.e., buffering agents) and other coatings well known in the pharmaceutical formulation art. They can optionally contain opacifying agents and can also be compositions that release the active ingredient only, or preferably only, in a certain part of the intestine, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0398] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. Besides the active compound, the liquid dosage forms may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents, and emulsifying agents such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3 - butanediol, dimethylformamide, oils (especially cottonseed, peanut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols, and fatty acid esters of sorbitan and mixtures thereof. Besides the inert diluents, the oral compositions may also include adjuvants such as wetting agents, emulsifying agents, and suspending agents, sweetening agents, flavoring agents, and perfuming agents.

[0399] Alternatively, the pharmaceutically acceptable compositions described herein can be administered in the form of suppositories for rectal or vaginal administration. These can be prepared by mixing the compounds of the present disclosure with suitable non-irritating excipients or carriers which are solid at room temperature but liquid at body temperature (e.g., rectal or vaginal temperature), and will thus melt in the rectal or vaginal cavity to release the active compound. Such materials include cocoa butter, suppository waxes (e.g., beeswax), and polyethylene glycols.

[0400] The pharmaceutically acceptable compositions described herein can also be administered topically, particularly when the treatment target includes areas or organs that are readily accessible by topical administration, including diseases of the eye, skin, or lower intestine. Topical administration to the lower intestine can be achieved by rectal suppository formulations (see above) or suitable enema formulations.

[0401] Dosage forms for topical or transdermal administration of the provided compounds include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active ingredient is mixed with a pharmaceutically acceptable carrier and any desired preservatives or buffering agents, if required, under sterile conditions. Ophthalmic formulations, ear drops, and eye drops are also contemplated within the scope of the present disclosure. Additionally, the present disclosure contemplates the use of transdermal patches, which have the additional advantage of delivering the compound to the body in a controlled manner. Such dosage forms can be made by dissolving or dispersing the compound in a suitable medium. Penetration enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0402] For topical administration, the provided pharmaceutically acceptable compositions can be formulated as suitable ointments containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of the present disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide compounds, emulsifying wax, and water. Alternatively, the provided pharmaceutically acceptable compositions can be formulated as suitable lotions or creams containing the 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 esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water.

[0403] For ophthalmic use, the provided pharmaceutically acceptable compositions can be formulated as a micronized suspension in isotonic, pH-adjusted sterile physiological saline, or preferably as a solution in isotonic, pH-adjusted sterile physiological saline, which may or may not contain preservatives such as benzalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutically acceptable compositions can be formulated in an ointment such as petrolatum.

[0404] The pharmaceutically acceptable compositions of the present disclosure can also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulations and can incorporate benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents and are prepared in the form of solutions in physiological saline.

[0405] Most preferably, the pharmaceutically acceptable compositions of the present disclosure are formulated for oral administration.

[0406] Identification and / or characterization of a compound and / or composition

[0407] Among other things, the present disclosure provides various techniques for identifying and / or characterizing the compounds and / or compositions described herein. For example, the present disclosure provides various assays for assessing SARM1 inhibitory activity, particularly for assessing SARM1 inhibitory activity.

[0408] 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 can be the absence of the relevant compound or composition. Alternatively or additionally, in some embodiments, the reference can be the presence of an alternative compound or composition, e.g., the alternative compound or composition has known performance in the relevant assay (e.g., as a positive or negative control, as understood in the art). In some embodiments, the reference can be a set of alternative but comparable conditions (e.g., temperature, pH, salt concentration, etc.). In some embodiments, the reference can be the performance of a compound or composition relative to a SARM1 variant.

[0409] Further alternatively or additionally, in some embodiments, in the assays described herein, the performance of one or more compounds or compositions of interest can be evaluated in the presence of a suitable reference compound or composition, e.g., to determine the ability of the compound or composition to compete with the reference.

[0410] In some embodiments, multiple compounds or compositions of interest can be analyzed and / or compared to the same reference in a particular assay. In some embodiments, such multiple compounds or compositions can be or include a set of compounds or compositions that are considered a "library" because the multiple members share one or more characteristics (e.g., structural elements, source characteristics, synthetic similarities, etc.).

[0411] Certain exemplary assays useful in the practice of the present disclosure are illustrated in the examples below. Those skilled in the art reading the present disclosure will recognize that useful or relevant systems for identifying and / or characterizing compounds and / or compositions according to the present disclosure are not limited to those included in the examples, or those otherwise discussed below.

[0412] In some embodiments, compounds and / or compositions can be determined and / or characterized based on one or more activities or features, such as promoting axonal integrity, cytoskeletal stability, and / or neuronal survival. In some embodiments, the provided SARM1 inhibitors inhibit the catabolism of NAD+ by SARM1. In some embodiments, the provided SARM1 inhibitors slow the rate of NAD+ catabolism.

[0413] In some embodiments, the provided SARM1 inhibitors reduce or inhibit the binding of SARM1 to NAD+. In some embodiments, the provided SARM1 inhibitors bind to SARM1 within a pocket that includes one or more catalytic residues (e.g., the catalytic cleft of SARM1). Examples of such catalytic residues include glutamate at position 642 (E642).

[0414] In some embodiments, the provided SARM1 inhibitors disrupt and / or prevent the polymerization of the TIR1 domain of SARM1. In some embodiments, the provided SARM1 inhibitors disrupt the polymerization of the SAM domain. In some embodiments, the provided SARM1 inhibitors disrupt the axonal signaling cascade that leads to NAD+ depletion.

[0415] In some embodiments, the present disclosure provides assays for one or more activities and / or features useful for identifying and / or characterizing compounds and / or compositions of interest. For example, in some embodiments, the present disclosure provides in vitro, cellular, and / or in vivo systems for assessing one or more such activities and / or features.

[0416] SARM1 activity assay

[0417] In some embodiments, a method of identifying a SARM1 inhibitor 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 the NAD+ in the mixture after incubation; and d) identifying the candidate inhibitor compound as an inhibitor if the amount of NAD+ is greater than the amount in a control mixture that does not include the candidate inhibitor.

[0418] In some embodiments, methods for identifying SARM1 inhibitors are provided, including: a) providing a mixture comprising 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 the molar ratio of a control mixture without the candidate inhibitor.

[0419] In some embodiments, methods for identifying SARM1 inhibitors are provided, including: a) providing a mixture comprising a solid support bound to i) full-length SARM1 and at least one tag, ii) NAD+, and iii) a candidate inhibitor; b) incubating the mixture; c) quantifying NAD+ after incubation; and d) identifying the candidate inhibitor compound as a SARM1 inhibitor if the concentration of NAD+ is greater than the concentration of a control.

[0420] SARM1 binding assay

[0421] In some embodiments, the efficacy of the provided SARM1 inhibitors can be determined according to assays described, for example, in WO 2018 / 057989, published on March 29, 2018, which is incorporated herein by reference in its entirety. In some embodiments, the provided SARM1 inhibitors can be applied to a solution containing SARM1 or a fragment thereof. In some embodiments, the provided SARM1 inhibitors can be applied to an in vitro system. In some embodiments, the provided SARM1 inhibitors can be applied to an in vivo system. In some embodiments, the provided SARM1 inhibitors can be applied to a patient. In some embodiments, the SARM1 inhibitors can be mixed with SARM1 or a fragment thereof that has been labeled with an epitope tag. In some embodiments, the amount of bound SARM1 inhibitor can be compared with the amount of unbound SARM1 inhibitor to determine the affinity for the SARM1 inhibitor.

[0422] In some embodiments, the mutant or fragment of SARM1 is a constitutively active SAM-TIR fragment. Constitutively active SARM1 fragments include, for example but not limited to, SARM1 lacking the autoinhibitory domain; at least one point mutation of SARM1 that inactivates the autoinhibitory domain; SARM1 fragments containing the TIR domain; or SARM1 fragments consisting of the SAM and TIR domains. In some embodiments, the SARM1 polypeptide 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 SARM1 polypeptide may include tags at the amino terminus, carboxyl terminus, or combinations thereof. In some embodiments, SARM1 or its fragment labeled with an epitope tag can be used to measure the binding efficacy of the provided SARM1 inhibitor.

[0423] Purification of the SARM1-TIR domain

[0424] In some embodiments, the SARM1-TIR domain can be engineered with various protein tags or epitope tags that can be used, for example, for purification. In some embodiments, the present disclosure also provides an NRK1-HEK293T cell line comprising HEK293T cells transformed with nicotinamide riboside kinase 1 (NRK1). In some embodiments, HEK293T cells are transformed or transfected with a DNA sequence encoding nicotinamide riboside kinase 1 (NRK1). In some embodiments, the DNA encoding NRK1 can be genomic or cDNA. In some embodiments, HEK293T cells are stably or transiently transfected with DNA encoding NRK1 that is exogenous to the host cell. In some embodiments, HEK293T cells are stably or transiently transfected with DNA encoding NRK1 such that the cells express NRK1 at elevated levels 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 sequences is a non-naturally occurring combination. In some embodiments, the DNA encoding NRK1, whether genomic or cDNA, comprises an expression vector, such as an FCIV expression vector. In some embodiments, the DNA encoding NRK1 is derived from genomic DNA or cDNA from a vertebrate or invertebrate species, such as but not limited to human, mouse, zebrafish, or Drosophila. In some configurations, the NRK1 DNA is human NRK1 DNA.

[0425] Applications and uses

[0426] The present disclosure provides various uses and applications of the compounds and / or compositions described herein, such as based on the activities and / or characteristics of these compounds and / or compositions as described herein. In some embodiments, such uses may include therapeutic and / or diagnostic uses. Alternatively, in some embodiments, such uses may include research, production, and / or other technical uses.

[0427] In one aspect, the present disclosure provides methods comprising administering to a subject one or more compounds of Formula I or Formula II, 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 or Formula II is a SARM1 inhibitor.

[0428] Another embodiment of the present disclosure relates to a method of inhibiting SARM1 activity in a patient, comprising the step of administering to the patient the provided compound or a composition comprising the compound.

[0429] Inhibiting an enzyme in a biological sample is useful for a variety of purposes known to those of skill in the art. Examples of such purposes include, but are not limited to, bioassays, gene expression studies, and biological target identification.

[0430] In certain embodiments, the present disclosure relates to a method of treating axonal degeneration in a biological sample, comprising the step of contacting the biological sample with a compound or composition of Formula I or Formula II. In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, in a method of inhibiting neuronal degeneration derived from a subject. In some embodiments, one or more of the compounds and / or compositions described herein can be used to inhibit the degeneration of neurons or a portion thereof in vitro. In some embodiments, one or more of the compounds and / or compositions described herein can be used as a stabilizer to promote neuronal survival in vitro.

[0431] In some embodiments, the provided compounds and / or compositions inhibit the NAD enzymatic activity of SARM1. Alternatively or additionally, in some embodiments, the provided compounds mitigate one or more attributes of neurodegeneration. In some embodiments, the present disclosure provides methods of treating neurodegenerative diseases or disorders associated with axonal degeneration.

[0432] In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, in medical practice. In some embodiments, one or more of the compounds and / or compositions described herein can 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 of the compounds and / or compositions described herein can be used, for example, to inhibit axonal degeneration, including axonal degeneration caused by NAD+ reduction or depletion. In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, to prevent axonal degeneration distal to axonal injury.

[0433] In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, in a method for inhibiting the degeneration of neurons of the peripheral nervous system or a part thereof. In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, in a method for inhibiting or preventing the degeneration of the central nervous system (neurons) or a part thereof. In some embodiments, one or more of the compounds or compositions described herein are characterized in that, when administered to a population of subjects, they reduce one or more symptoms or characteristics of neurodegeneration. For example, in some embodiments, the relevant symptoms or characteristics can be selected from the group consisting of the degree, rate, and / or timing of neuronal destruction.

[0434] In certain embodiments, the present disclosure provides compounds according to the present disclosure for use, for example, as analytical tools, as probes in biological assays, or as therapeutic agents. The compounds provided by the present disclosure can also be used to study SARM1 activity in biological and pathological phenomena and for comparative evaluation of new SARM1 activity inhibitors in vitro or in vivo. In certain embodiments, the present 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 vector constructs and / or polypeptides) that can be used to assay SARM1 activity. For example, in some embodiments, the provided assays can utilize, for example, SAM-TIR lacking the SARM1 N-terminal autoinhibitory domain, and / or one or more tagged versions of the TIR domain.

[0435] In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, in a method for inhibiting the degeneration of neurons derived from a subject. In some embodiments, one or more of the compounds and / or compositions described herein can be used to inhibit the degeneration of neurons or a part thereof cultured in vitro. In some embodiments, one or more of the compounds and / or compositions described herein can be used as stabilizers to promote the survival of neurons in vitro.

[0436] In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, to affect biomarkers associated with neurodegeneration. In some embodiments, changes in biomarkers can be detected systemically or with samples from cerebrospinal fluid (CSF), plasma, serum, and / or tissue from a subject. In some embodiments, one or more of the compounds and / or compositions can be used to affect changes in the concentration of neurofilament light chain (NF-L) and / or neurofilament heavy chain (NF-H) contained in the cerebrospinal fluid of a subject. In some embodiments, one or more of the compounds and / or compositions described herein can affect constitutive NAD and / or cADPR levels in neurons and / or axons.

[0437] In some embodiments, one or more of the compounds and / or compositions described herein can affect detectable changes in the levels of one or more neurodegeneration-related proteins in a subject. 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, neurogranin, neuron-specific enolase (NSE), soluble amyloid precursor protein (sAPP)α, sAPPβ, soluble triggering receptor expressed on myeloid cells (sTREM)2, phosphorylated tau, and / or total tau. In some embodiments, one or more of the compounds and / or compositions described herein can affect changes in cytokines and / or chemokines, including but not limited to Ccl2, Ccl7, Ccl12, Csf1, and / or Il6.

[0438] Diseases, disorders and conditions

[0439] In some embodiments, the compounds and / or compositions described herein can be administered to a subject having one or more diseases, disorders, or conditions.

[0440] In some embodiments, the condition is an acute condition. In some embodiments, the condition is a chronic condition.

[0441] In some embodiments, the condition is characterized by axonal degeneration of the central nervous system, peripheral nervous system, optic nerve, cranial nerves, or a combination thereof.

[0442] In some embodiments, the condition is or includes an acute injury to the central nervous system, such as an injury to the spinal cord and / or traumatic brain injury. In some embodiments, the condition is or includes a chronic injury to the central nervous system, such as an injury to the spinal cord, traumatic brain injury, and / or traumatic axonal injury. In some embodiments, the condition is or includes chronic traumatic encephalopathy (CTE).

[0443] In some embodiments, the condition is a chronic condition affecting the central nervous system, such as Parkinson's disease, amyotrophic lateral sclerosis, multiple sclerosis, or Huntington's disease, Alzheimer's disease.

[0444] In some embodiments, the condition is an acute peripheral neuropathy. Chemotherapy-induced peripheral neuropathy (CIPN) is an example of an acute peripheral neuropathy. CIPN can be associated with a variety of drugs, such as but not limited to thalidomide, epothilone (e.g., ixabepilone), taxane (e.g., paclitaxel and docetaxel), vinca alkaloid (e.g., vinblastine, vinorelbine, vincristine, and vindesine), proteasome inhibitor (e.g., bortezomib), platinum drugs (e.g., cisplatin, oxaliplatin, and carboplatin).

[0445] In some embodiments, the condition is a chronic condition affecting the peripheral nervous system, such as diabetic neuropathy, HIV neuropathy, Charcot Marie Tooth disease, or amyotrophic lateral sclerosis.

[0446] In some embodiments, the condition is an acute condition affecting the optic nerve, such as acute optic neuropathy (AON) or acute angle-closure glaucoma.

[0447] In some embodiments, the condition is a chronic condition affecting the optic nerve, such as Leber congenital amaurosis, Leber hereditary optic neuropathy, primary open-angle glaucoma, and autosomal dominant optic atrophy.

[0448] In some embodiments, one or more of the compounds and / or compositions described herein can be used, for example, to treat one or more neurodegenerative diseases, disorders, or conditions selected from the group consisting of neuropathy or axonopathy. In some embodiments, one or more of the compounds and / or compositions described herein can 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 de novo or somatic mutations. In some embodiments, the neuropathy associated with axonal degeneration is selected from the list included herein. In some embodiments, the neuropathy or axonopathy associated with axonal degeneration includes, but is not limited to, Parkinson's disease, non-Parkinson's disease, Alzheimer's disease, herpes infection, diabetes, amyotrophic lateral sclerosis, demyelinating diseases, ischemia or stroke, chemical injury, thermal injury, and AIDS.

[0449] In some embodiments, one or more of the compounds or compositions described herein are characterized in that, when administered to a population of subjects, they reduce one or more symptoms or features of neurodegeneration. For example, in some embodiments, the relevant symptoms or features can be selected from the group consisting of the degree, rate, and / or timing of neuronal destruction. In some embodiments, neuronal destruction can be or include axonal degeneration, synaptic loss, dendritic loss, loss of synaptic density, loss of dendritic arborization, loss of axonal branching, loss of neuronal density, loss of myelination, loss of neuronal cell bodies, loss of synaptic potentiation, loss of action potential potentiation, loss of cytoskeletal stability, loss of axonal transport, loss of ion channel synthesis and turnover, loss of neurotransmitter synthesis, loss of neurotransmitter release and reuptake ability, loss of axonal potential propagation, neuronal hyperexcitability, and / or neuronal hypoexcitability. In some embodiments, neuronal destruction is characterized by the inability to maintain an appropriate resting neuronal membrane potential. In some embodiments, neuronal destruction is characterized by the presence of inclusion bodies, plaques, and / or neurofibrillary tangles. In some embodiments, neuronal destruction is characterized by the presence of stress granules. In some embodiments, neuronal destruction is characterized by the intracellular activation of one or more members of the cysteine-aspartic protease (Caspase) 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.

[0450] In some embodiments, a neurodegenerative or neurological disease or disorder is associated with axonal degeneration, axonal injury, axonal pathology, demyelinating diseases, central pontine myelinolysis, nerve injury diseases or disorders, metabolic diseases, mitochondrial diseases, metabolic axonal degeneration, axonal injury caused by leukodystrophy or leukodystrophy. In some embodiments, the neurodegenerative or neurological disease or disorder is selected from the group consisting of: spinal cord injury, stroke, multiple sclerosis, progressive multifocal leukoencephalopathy, congenital hypomyelination, encephalomyelitis, acute disseminated encephalomyelitis, central pontine myelinolysis, osmotic hyponatremia, hypoxic demyelination, ischemic demyelination, adrenoleukodystrophy, Alexander's disease, Niemann-Pick disease, Pelizaeus-Merzbacher disease, periventricular leukomalacia, globoid cell leukodystrophy (Krabbe's disease), Wallerian degeneration, optic neuritis, transverse myelitis, amyotrophic lateral sclerosis (ALS, Lou Gehrig's disease), Huntington's disease, Alzheimer's disease, Parkinson's disease, Tay-Sacks disease, Gaucher's disease, Hurler Syndrome, traumatic brain injury, post-radiation injury, neurological complications of chemotherapy (chemotherapy-induced neuropathy; CIPN), neuropathy, acute ischemic optic neuropathy, vitamin B 12Deficiency diseases, single vitamin E deficiency syndrome, Bassen-Kornzweig syndrome, glaucoma, Leber's hereditary optic neuropathy (neuropathy), Leber's congenital amaurosis, neuromyelitis optica, metachromatic leukodystrophy, acute hemorrhagic leukoencephalitis, trigeminal neuralgia, Bell's palsy, cerebral ischemia, multiple system atrophy, traumatic glaucoma, HTLV-1 associated myelopathy, West Nile virus encephalopathy, La Crosse virus encephalitis, Bunyavirus encephalitis, viral encephalitis in children, essential tremor, Charcot-Marie-Tooth disease, motor neuron disease, spinal muscular atrophy (SMA), hereditary sensory and autonomic neuropathy (HSAN), adrenomyeloneuropathy, progressive supranuclear palsy (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, Guillain-Barre syndrome, severe acute motor axonal neuropathy (AMAN), Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, spinocerebellar ataxia, preeclampsia, hereditary spastic paraplegia, spastic paraplegia, familial spastic paraplegia, French settlement disease, Strumpell-Lorrain disease, and non-alcoholic steatohepatitis (NASH).

[0451] In some embodiments, the present disclosure provides SARM1 activity inhibitors for treating neurodegenerative or neurological diseases or disorders involving axonal degeneration or axonopathy. The present disclosure also provides methods of using SARM1 activity inhibitors to treat, prevent, or improve axonal degeneration, axonopathy, and neurodegenerative or neurological diseases or disorders involving axonal degeneration.

[0452] In some embodiments, the present disclosure provides methods for treating neurodegenerative or neurological diseases or disorders associated with axonal degeneration, axonal injury, axonal pathologies, demyelinating diseases, central pontine myelinolysis, nerve injury diseases or disorders, metabolic diseases, mitochondrial diseases, metabolic axonal degeneration, axonal injury caused by leukodystrophy or leukodystrophy.

[0453] In some embodiments, neuropathies and axonal pathologies include any disease or condition involving neurons and / or supporting cells, such as glial cells, muscle cells, or fibroblasts, particularly those involving axonal injury. Axonal injury can be caused by traumatic injury or non-mechanical injury caused by disease, condition, or exposure to toxic molecules or drugs. The result of such injury can be axonal degeneration or dysfunction and loss of functional neuronal activity. The diseases and conditions resulting from or associated with such axonal injury are one of many neurological diseases and conditions. Such neuropathies can include peripheral neuropathies, central neuropathies, and combinations thereof. In addition, peripheral neurological manifestations can be mainly caused by diseases concentrated in the central nervous system, while central nervous system manifestations can be substantially caused by peripheral or systemic diseases.

[0454] In some embodiments, peripheral neuropathy can involve injury to the peripheral nerves and / or can be caused by diseases of the nerves or due to systemic diseases. Some such diseases can 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 embodiments, peripheral nerve degeneration is caused by traumatic (mechanical) injury to the nerve as well as chemical or thermal injury to the nerve. Such conditions that damage the peripheral nerves include compression or pinching, such as glaucoma, carpal tunnel syndrome, direct trauma, penetrating injury, contusion, fracture, or bone dislocation; pressure on superficial nerves (ulnar, radial, or peroneal nerves), which may be due to long-term use of crutches or staying in one position for too long, or due to tumors; intraneural hemorrhage; ischemia; exposure to cold or radiation or certain drugs or toxic substances, such as herbicides or pesticides. In particular, nerve injury may be due to chemical injury by cytotoxic anticancer agents, such as paclitaxel, cisplatin, proteasome inhibitors, or vinca alkaloids such as vincristine. Typical symptoms of such 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 embodiments, neuropathy is associated with mitochondrial dysfunction. Such neuropathy can exhibit a decrease in energy levels, i.e., a decrease in NAD and ATP levels.

[0455] In some embodiments, peripheral neuropathy is a metabolic and endocrine neuropathy that includes a broad range of peripheral nerve disorders associated with systemic diseases of metabolic origin. These diseases include, for example, diabetes, hypoglycemia, uremia, hypothyroidism, liver failure, polycythemia, amyloidosis, acromegaly, porphyria, lipid / glycolipid metabolic disorders, nutritional / vitamin deficiencies, and mitochondrial disorders, among others. The common feature of these diseases is that they affect the peripheral nerves due to structural or functional changes in the myelin sheath and axons resulting from dysregulation of metabolic pathways.

[0456] In some embodiments, neuropathy includes optic neuropathy such as glaucoma; retinal ganglion degeneration such as that associated with retinitis pigmentosa and extraretinal neuropathy; optic neuritis and / or degeneration, including that associated with multiple sclerosis; traumatic injury to the optic nerve, which can 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 that secondary to giant cell arteritis; metabolic optic neuropathy such as neurodegenerative diseases, including the aforementioned Leber's neuropathy, nutritional deficiencies such as vitamin B12 or folate deficiency, and toxicity such as that due to ethambutol or cyanide; neuropathy caused by adverse drug reactions and neuropathy caused by vitamin deficiencies. Ischemic optic neuropathy also includes non-arteritic anterior ischemic optic neuropathy.

[0457] In some embodiments, neurodegenerative diseases associated with neuropathy or axonopathy in the central nervous system include a variety of diseases. Such diseases include those involving progressive dementia, such as Alzheimer's disease, senile dementia, Pick's disease, and Huntington's disease; central nervous system diseases that affect muscle function, such as Parkinson's disease, motor neuron diseases, and progressive ataxia, such as amyotrophic lateral sclerosis; demyelinating diseases, such as multiple sclerosis; viral encephalitis, such as that caused by enteroviruses, arboviruses, and herpes simplex virus; and prion diseases. Mechanical injuries, such as glaucoma or traumatic injury to the head and spine, can also cause nerve damage and degeneration in the brain and spinal cord. In addition, ischemia and stroke, as well as conditions such as nutritional deficiencies and chemical toxicity (such as chemotherapeutic agents), can cause central nervous system neuropathy.

[0458] In some embodiments, the present disclosure provides a method for treating neuropathy or axonopathy associated with axonal degeneration. In some such embodiments, the neuropathy or axonopathy associated with axonal degeneration can be any one of a variety of neuropathies or axonopathies, such as those that are genetic or congenital or associated with Parkinson's disease, Alzheimer's disease, herpes infection, diabetes, amyotrophic lateral sclerosis, demyelinating diseases, ischemia or stroke, chemical injury, thermal injury, and AIDS-related neuropathies or axonopathies. In addition, neurodegenerative diseases not mentioned above and subsets of the diseases mentioned above can also be treated by the methods of the present disclosure. Such subsets of diseases can include Parkinson's disease or non-Parkinson's disease, or Alzheimer's disease.

[0459] Subject

[0460] In some embodiments, the compounds and / or compositions described herein are administered to a subject having or at risk of having a disease, disorder, or condition described herein; in some embodiments, such disease, disorder, or condition is characterized by axonal degeneration, such as one of the conditions mentioned herein.

[0461] In some embodiments, the subject to whom the compounds or compositions described herein are administered exhibits one or more signs or symptoms associated with axonal degeneration; in some embodiments, the subject does not exhibit any signs or symptoms of neurodegeneration.

[0462] In some embodiments, the provided method comprises administering a compound of formula I or formula II to a patient in need thereof. In some such embodiments, the patient is at risk of developing a condition characterized by axonal degeneration. In some embodiments, the patient has a condition characterized by axonal degeneration. In some embodiments, the patient has been diagnosed with a condition characterized by axonal degeneration.

[0463] In some embodiments, the provided method comprises administering the compositions described herein to a population of patients in need thereof. In some embodiments, the population is those individuals who are involved in activities with a high likelihood of traumatic neuronal injury. In some embodiments, the population is athletes who are involved in contact sports or other high-risk activities.

[0464] In some embodiments, the subject is at risk of developing a condition characterized by axonal degeneration. In some embodiments, the subject is identified as being at risk of axonal degeneration, such as 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.

[0465] In some embodiments, the patient is at risk of developing a neurodegenerative disorder. In some embodiments, the patient is an elderly individual. In some embodiments, the patient is known to have a genetic risk factor for neurodegeneration. In some embodiments, the patient has a family history of neurodegenerative disease. In some embodiments, the patient expresses one or more copies of a known genetic risk factor for neurodegeneration. In some embodiments, the patient is from a population with a high incidence of neurodegeneration. In some embodiments, the patient has a hexanucleotide repeat expansion in chromosome 9 open reading frame 72. In some embodiments, the patient has one or more copies of the ApoE4 allele.

[0466] In some embodiments, the subject to which the compounds or compositions described herein are administered can be or comprise a subject having or at risk of having a neurodegenerative disease, disorder or condition. In some embodiments, the neurodegenerative disease, disorder or condition can be or comprise traumatic neuronal injury. In some embodiments, the traumatic neuronal injury is blunt force trauma, closed head injury, open head injury, exposure to concussive and / or explosive forces, penetrating injury to the cranial cavity or body innervation regions. In some embodiments, the traumatic neuronal injury is a force that causes axonal deformation, stretching, compression or shearing.

[0467] In some embodiments, the subject engages in activities identified as risk factors for neuronal degeneration, e.g., a subject who engages in contact sports or a profession with a high probability of traumatic neuronal injury.

[0468] For example, the subject can be a patient who is receiving or has been prescribed chemotherapy associated with peripheral neuropathy. Examples of chemotherapeutic agents include, but are not limited to, thalidomide, epothilones (e.g., ixabepilone), taxanes (e.g., paclitaxel and docetaxel), vinca alkaloids (e.g., vinblastine, vinorelbine, vincristine and vindesine), proteasome inhibitors (e.g., bortezomib), platinum drugs (e.g., cisplatin, oxaliplatin and carboplatin).

[0469] In some embodiments, the provided method comprises administering the compositions 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 comprises monitoring the levels of the biomarkers in the patient or patient population and adjusting the dosing regimen accordingly.

[0470] Administration

[0471] 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 in the pharmaceutical compositions or regimens described herein can be selected by a medical practitioner and may vary for different subjects, for example after considering one or more of the species, age, and general condition of the subject, and / or the properties of the particular compound or composition, its mode of administration, etc. Alternatively, in some embodiments, the amount of a particular compound included in and / or delivered by administration in a pharmaceutical composition or regimen as described herein can be standardized within a relevant patient population (e.g., all patients, patients of a particular age or disease stage, or all patients expressing a particular biomarker, etc.).

[0472] The compounds or compositions provided by the present disclosure are preferably formulated in unit dosage forms to facilitate administration and uniformity of dosage. As used herein, the expression "unit dosage form" refers to physically discrete dosage units suitable for the patient to be treated. However, it should be understood that the total daily dosage of the compounds and compositions provided by the present disclosure will be determined by the attending physician within the scope of reasonable medical judgment. The specific effective dosage level for any particular patient or organism will depend on a variety of factors, including the disease being treated and the severity of the disease; the clinical condition of the individual patient; the cause of the disease; the activity of the particular compound employed; the particular composition employed; the age, weight, general health, sex, and diet of the patient; the time of administration of the particular compound, the site of delivery of the medicament, the route of administration, and the excretion rate; the duration of treatment; drugs used in combination with or concurrently with the particular compound employed, and similar factors well known in the medical arts. 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 to prevent or treat an undesirable disease or condition, such as neurodegeneration or traumatic nerve injury.

[0473] The pharmaceutically acceptable compositions of the present disclosure can be administered to humans and other animals orally, rectally, intravenously, parenterally, intracisternally, vaginally, intraperitoneally, topically (such as powders, ointments, or drops), buccally, as an oral or nasal spray, etc., depending on the severity of the disease, disorder, or infection being treated. In certain embodiments, the daily dose is given as a single daily dose or in divided doses two to six times a day, or in a sustained release form. This dosing regimen can be adjusted to provide an optimal therapeutic response. The compound can be administered according to a regimen of once to four times a day, preferably once or twice a day.

[0474] In some embodiments, the compositions of the present disclosure can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or by an implanted reservoir. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intraliver, intradermal, intraocular, intralesional, and intracranial injection or infusion techniques. Preferably, the composition is administered orally, intraperitoneally, or intravenously.

[0475] In some embodiments, the pharmaceutically acceptable compositions of the present disclosure can also be administered topically, particularly when the treatment target includes regions or organs that are readily accessible by topical administration, including diseases of the eye, skin, or lower intestine. Topical formulations suitable for each of these regions or organs are readily prepared.

[0476] Most preferably, the pharmaceutically acceptable compositions of the present disclosure are formulated for oral administration. Such formulations can be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions of the present disclosure are not administered with food. In other embodiments, the pharmaceutically acceptable compositions of the present disclosure are administered with food.

[0477] Those additional agents can be administered separately from the provided compound or its composition as part of a multi-dose regimen. Alternatively, those agents can be part of a single dosage form and mixed with the provided compound in a single composition. If administered as part of a multi-dose regimen, the two active agents can be provided simultaneously, sequentially, or at intervals of time (usually within five hours of each other) from one another.

[0478] It should also be understood that the specific dosage and treatment regimen for any particular patient may depend on a variety of factors, including the activity of the specific compound employed, age, weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. In some embodiments, the amount of the compound of the present disclosure in the composition will also depend on the specific compound in the composition.

[0479] In some embodiments, the SARM1 inhibitors described herein can be utilized in combination with one or more other therapies to treat related diseases, disorders, or conditions. In some embodiments, the dosage of the SARM1 inhibitor is altered when using combination therapy compared to when administered as monotherapy; alternatively or additionally, in some embodiments, the therapy administered in combination with the SARM1 inhibition described herein is administered according to a regimen or protocol different from that when administered alone or in combination with one or more therapies other than SARM1 inhibition. In some embodiments, the composition comprising the additional therapeutic agent, the additional therapeutic agent, and the provided compound can act synergistically. In some embodiments, one or both of the therapies used in the combination regimen are administered at a lower level or less frequently compared to when used as monotherapy.

[0480] In some embodiments, the compounds and / or compositions described herein are administered together with chemotherapeutic agents, including but not limited to alkylating agents, anthracyclines, taxanes, epothilones, histone deacetylase inhibitors, topoisomerase inhibitors, kinase inhibitors, nucleotide analogs, peptide antibiotics, platinum agents, retinoids, vinca alkaloids, and derivatives. In some embodiments, the compounds and / or compositions described herein are administered in combination with a PARP inhibitor.

[0481] Examples

[0482] This teaching includes the descriptions provided in the examples, which are not intended to limit the scope of any claim. Unless specifically stated in the past tense, those included in the examples are not intended to imply that the experiments were actually conducted. The following non-limiting examples are provided to further illustrate this teaching. Those skilled in the art will understand, in light of the present disclosure, that many changes can be made in the specific embodiments disclosed and still obtain similar or analogous results without departing from the spirit and scope of this teaching.

[0483] Methods

[0484] Some of the methods and compositions described herein utilize laboratory techniques well known to those of skill 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, N.Y., 2001; Methods In Molecular Biology, edited by Richard, Humana Press, NJ, 1995; Spector, D.L. et al., Cells: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1998; and Harlow, E., Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1999. Methods of administration and dosage regimens of drugs can be determined according to standard principles of pharmacology using methods provided in standard references such as Remington: the Science and Practice of Pharmacy (edited by Alfonso R. Gennaro, 19th ed., 1995); Hardman, J.G. et al., Goodman & Gilman's The Pharmacological Basis of Therapeutics, 9th ed., McGraw-Hill, 1996; and Rowe, R.C. et al., Handbook of Pharmaceutical Excipients, 4th ed., Pharmaceutical Press, 2003.

[0485] Example 1: Characterization of the Compound

[0486] LCMS Method:

[0487] Analytical LC / MS Analytical Method A:

[0488] ESI+ / - ion mode 100 - 1000 Da

[0489] Chromatographic column: XBridge C18, 50 x 4.6 mm, 3.5 µm

[0490] Temperature: 40 °C

[0491] Gradient:

[0492] Time (minutes) <![CDATA[Water (10 mM NH NH4HCO3)]]> Acetonitrile Flow rate (mL / minute) 0 95％ 5％ 2.0 1.2 5％ 95％ 2.0 3.0 5％ 95％ 2.0

[0493] Analytical LC / MS analysis method B:

[0494] ESI+ / - ion mode 150 - 850 Da

[0495] Chromatographic column: Phenomenex Kinetix-XB C18, part number 00D-4498-AN, 2.1 x 100 mm, 1.7 µm Temperature: 40 °C

[0496] Gradient:

[0497] Time (minutes) Water + 0.1% formic acid Acetonitrile + 0.1% formic acid Flow rate (mL / minute) 0.00 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

[0498] Analytical LC / MS analysis method C:

[0499] ESI+ / - ion mode 100 - 1000 Da

[0500] Chromatographic column: XBridge C18, 50 x 4.6 mm, 3.5 µm

[0501] Temperature: 40 °C

[0502] Gradient:

[0503] Time (minutes) Water + 0.1% TFA Acetonitrile + 0.1% TFA Flow rate (mL / minute) 0 95％ 5％ 1.8 1.8 5％ 95％ 1.8 3.0 5％ 95％ 1.8

[0504] The results are listed in Table 1:

[0505] Table 1.

[0506]

[0507]

[0508]

[0509]

[0510]

[0511]

[0512]

[0513]

[0514]

[0515]

[0516]

[0517]

[0518]

[0519]

[0520]

[0521]

[0522]

[0523]

[0524]

[0525]

[0526]

[0527]

[0528]

[0529]

[0530]

[0531]

[0532]

[0533]

[0534]

[0535] Example 2: SAM-TIR SARM1 IC50 Determination

[0536] This example describes the determination of SAM-TIR NAD enzyme activity and the use of this determination to measure the efficacy of compounds of formula I or formula II in blocking SARM1-mediated NAD+ cleavage. The assay was optimized to characterize the efficacy of compounds of formula I or formula II in inhibiting SARM1 activity and to calculate the IC50 value for each compound. The assay utilizes a fragment of the SARM1 molecule containing the SAM and TIR domains. As shown herein, expression of this fragment without the autoinhibitory N-terminal domain produces a constitutively active enzyme that cleaves NAD+.

[0537] Preparation of SARM1 SAM-TIR Lysate (STL)

[0538] NRK1-HEK293T cells refer to a cell line stably transfected with an FCIV expression vector that expresses human nicotinamide riboside kinase 1 (NRK1), an enzyme that converts the NAD+ biosynthetic precursor nicotinamide riboside (NR) to NMN, the direct precursor of NAD+. When NR is provided, this cell line increases intracellular NAD+ levels and maintains cell viability upon expression of SARM1 SAM-TIR. Figure 2 Shown is that the NRK1-HEK293T stable line with NR supplementation maintains higher NAD+ levels after SARM1-TIR expression. Data were generated from three independent NAD+ measurements of three independent transfection experiments and normalized to data from non-transfected experiments run in parallel. Data are represented as mean ± SEM; error bars: SEM; ***P < 0.001 two-tailed Student's t-test.

[0539] NRK1-HEK293T cells represent a cell line stably transfected with an FCIV expression vector that expresses human nicotinamide riboside kinase 1 (NRK1), an enzyme that converts the NAD+ biosynthetic precursor nicotinamide riboside (NR) to NMN, the direct precursor of NAD+.

[0540] Seed NRK1-HEK293T cells at 20×10 6 cells per plate into 150 cm 2 plates. The next day, transfect the cells with 15 μg of FCIV-SST (SAM-TIR expression plasmid, SEQ ID NO:1) using X-TREMEGENE TM 9 DNA transfection reagent (Roche product #06365787001).

[0541]

[0542] At the time of transfection, the culture was supplemented with 1 mM NR to minimize the toxicity of SAM-TIR overexpression. Forty-eight hours after transfection, the cells were harvested, pelleted by centrifugation at 1,000 rpm (Sorvall ST 16R centrifuge, Thermo Fisher), and washed once with cold PBS (0.01 M phosphate-buffered saline, 0.138 M NaCl; 0.0027 M KCl; pH 7.4). The cells were resuspended in PBS with protease inhibitor (cOmplete TM Protease Inhibitor Cocktail, Roche product #11873580001) and cell lysates were prepared by sonication (Branson Sonifer 450, output = 3, 20 pulses). The lysates were centrifuged (12,000×g at 4 °C for 10 minutes) to remove cell debris, and the supernatant (containing SARM1 SAM-TIR protein) was stored at -80 °C for subsequent in vitro SARM1 SAM-TIR NADase assays (see below). Protein concentration was determined by the bicinchoninic acid (BCA) method and used to normalize the lysate concentration.

[0543] Determination of SAM-TIR IC50 of the compound represented by Formula I or Formula II

[0544] The enzymatic assay was carried out in a 384-well polypropylene plate in Dulbecco's PBS buffer with a final assay volume of 20 μL. The SAM-TIR lysate at a final concentration of 5 μg / mL was pre-incubated with the corresponding compound at a final assay concentration of 1% DMSO for 2 hours at room temperature. The reaction was initiated by adding NAD+ at a final assay concentration of 5 μM as a substrate. After incubation at room temperature for 2 hours, the reaction was terminated with 40 μL of a termination solution of 7.5% trichloroacetic acid in acetonitrile. The concentrations of NAD+ and ADPR were analyzed by a RapidFire high-throughput mass spectrometry system (Agilent Technologies, Santa Clara, CA) using an API4000 triple quadrupole mass spectrometer (AB Sciex, Framingham, MA).

[0545] The results are listed in Table 2 below. The IC 50 of the compound with the designated "A" activity is < 1 μM; the IC 50 of the compound with the designated "B" activity is 1 - 5 μM; the IC 50 of the compound with the designated "C" activity is > 5 μM.

[0546] Table 2.

[0547]

[0548]

[0549]

[0550]

[0551]

[0552] Example 3: Axonal Degeneration Index

[0553] This example illustrates the in vitro axonal degeneration assay for characterizing the compounds shown in Formula I or Formula II. This assay is used to test the efficacy of the compounds shown in Formula I or Formula II in preventing axonal degeneration in mouse dorsal root ganglion (DRG) hanging drop cultures.

[0554] Mouse DRG hanging drop culture: Mouse dorsal root ganglion neurons (DRGs) were dissected from E12.5 CD1 mice (50 ganglia per embryo) and incubated in 0.5% trypsin solution containing 0.02% EDTA (Gibco) at 37 °C for 15 minutes. The cells were then gently triturated by pipetting and washed three times with DRG growth medium (Neurobasal medium (Gibco) containing 2% B27 (Invitrogen), 100 ng / ml 2.5S NGF (Harland Bioproducts), 1 mM 5-fluoro-2'-deoxyuridine (Sigma), penicillin, and streptomycin). The cells were suspended in DRG growth medium. 5000 cells / well were spotted 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) to create DRG hanging drop cultures. The cells were allowed to adhere to the plate in a humidified tissue culture incubator (5% CO2) for 15 minutes, followed by gently adding DRG growth medium (100 μl per well).

[0555] Axonal degeneration assay: Axonal degeneration was induced by manual axotomy using a scalpel or chemical toxic stimulation. After an appropriate experimental period, the DRG cultures were fixed in 1% PFA plus sucrose and stored in the refrigerator before imaging. Bright-field images of DRG axons and cell bodies were collected using a 20× water immersion lens of a Phenix automated confocal microscope (PerkinElmer), and axons were quantified using an in-house developed script (Acapella, PerkinElmer).

[0556] The results are listed in Table 3 below. The compounds described herein showed protection against axon breakage in cell assays and were sorted at IC 50 10 - 30 μM (B), < 10 μM (A).

[0557] Table 3.

[0558] Examples <![CDATA[Axonal Degeneration IC 50 > 6 B 7 B 27 A 12 A 31 B

Claims

"1. A compound having formula II: or a pharmaceutically acceptable salt thereof, wherein R 1 selected from -CN, -NO2, -C(O)R, -S(O)2R, -CON(R)2, -S(O)2N(R)2, -CO2R, -C(=NR)N(R)2 and a 5-membered heteroaryl ring having 2-3 heteroatoms independently selected from nitrogen, oxygen and sulfur; R 2 is -R; R 3 is -(CH2) 0-2 Cy, or: R 2 and R 3 together with the nitrogen atom to which they are attached form a 4- to 7-membered saturated or partially unsaturated ring fused to Cy, or form a 4- to 7-membered saturated or partially unsaturated ring substituted with -Cy; Cy is selected from phenyl, a 3- to 7-membered saturated or partially unsaturated carbocyclic ring, a 5- to 6-membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, an 8- to 10-membered bicyclic heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and an 8- to 10-membered bicyclic aryl ring, wherein each phenyl, carbocyclic ring, heteroaryl ring and aryl ring is substituted with 0-4 R x substituted; Each R x is independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R, an optionally substituted C 1-6 aliphatic group, an optionally substituted 5- to 6-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur, and an optionally substituted 8- to 10-membered heteroaryl ring having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur; R 4 is -R; and Each R is independently hydrogen, optionally substituted C 1-6 aliphatic, optionally substituted phenyl, or optionally substituted 3- to 7-membered saturated or partially unsaturated carbocycle, or: two instances of R together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocycle.

2. The compound according to claim 1, wherein the compound has formula I: or a pharmaceutically acceptable salt thereof, wherein R 1 selected from -CN, -NO2, -C(O)R, -S(O)2R, -CON(R)2, -S(O)2N(R)2 and -CO2R; R 2 is - R; R 3 is -(CH2) 0-2 Cy, or: R 2 and R 3 together with the nitrogen to which they are attached form a 4- to 7-membered saturated or partially unsaturated ring fused to Cy, or form a 4- to 7-membered saturated or partially unsaturated ring substituted by -Cy; Cy is selected from phenyl, a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, an 8- to 10-membered bicyclic heteroaryl ring having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and an 8- to 10-membered bicyclic aryl ring, wherein each phenyl, heteroaryl ring and aryl ring is substituted with 0 to 4 -R x substituted; Each -R x is independently selected from halogen, -CN, -NO2, -OR, -SR, -N(R)2, -SO2R, -SO2N(R)2, -CO2R, -CON(R)2, -N(R)SO2R, -N(R)C(O)R and optionally substituted C 1-6 aliphatic; R 4 is -R; Each R is independently hydrogen or optionally substituted C 1-6 aliphatic; or: two instances of R together with the atoms to which they are attached form a 3- to 6-membered saturated or partially unsaturated heterocycle.

3. The compound according to claim 1, wherein R 1 selected from -CN, -CON(R)2 and -C(=NR)N(R)2; and R 2 Selected from -H and -CH3.

4. The compound according to claim 1, wherein: R 1 selected from -CN, -CON(R)2 and -C(=NR)N(R)2; R 2 selected from -H and -CH3; R 3 is - Cy; and - Cy is selected from 5. The compound according to claim 4, wherein: R 1 is -CN; and R 2 is - H.

6. The compound according to claim 4, wherein: R 1 is -CONH2; and R 2 is - H.

7. A method comprising the steps of: administering a compound according to any one of claims 1 to 6 to (i) a subject having a condition characterized by axonal degeneration or (ii) a subject at risk of developing a condition characterized by axonal degeneration.

8. A method of treating or preventing axonal degeneration, comprising administering to a subject in need thereof a compound according to any one of claims 1 to 6.

9. A method of inhibiting SARM1, comprising contacting a biological sample with a compound according to any one of claims 1 to 6.

10. The method according to claim 9, wherein SARM1 is inhibited by covalently modifying a cysteine residue of SARM1.

11. The method according to claim 10, wherein the cysteine residue of SARM1 is Cys635.

12. The method according to claim 10, wherein the cysteine residue of SARM1 is Cys629.

13. The method according to claim 10, wherein the cysteine residue of SARM1 is Cys649.

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