CN118946553A

CN118946553A - SARM1 inhibitor compound, pharmaceutical composition comprising the same, and preparation method and use thereof

Info

Publication number: CN118946553A
Application number: CN202380032148.0A
Authority: CN
Inventors: 潘伟; 刘佳乐; 鄂镜雯; 史才遵; 马洪艳
Original assignee: Shenzhen Zhongge Biotechnology Co ltd
Current assignee: Shenzhen Zhongge Biotechnology Co ltd
Priority date: 2022-04-08
Filing date: 2023-04-07
Publication date: 2024-11-12

Abstract

The present disclosure provides SARM1 inhibitor compounds of formula (I), pharmaceutical compositions comprising the same, methods of making and uses thereof. The compounds are useful for inhibiting SARM1 and/or for treating and/or preventing a disease, disorder, or condition characterized by axonal degeneration.

Description

SARM1 inhibitor compound, pharmaceutical composition comprising the same, and preparation method and use thereof

The present application claims priority from chinese patent application CN2022103687829, with application date 2022, 4, 8. The present application incorporates the entirety of the above-mentioned chinese patent application.

Technical Field

The present disclosure provides SARM1 inhibitor compounds of formula I, pharmaceutical compositions comprising the same, methods of making and uses thereof. The compounds are useful for inhibiting SARM1 and/or for treating and/or preventing a disease, disorder, or condition characterized by axonal degeneration.

Background

Axonal degeneration is a hallmark of a variety of neurological conditions including peripheral neuropathy, traumatic brain injury, and neurodegenerative diseases (Gerdts et al, SARM1activation is triggered locally by NAD (+) destruction (SARM 1activation triggers axon degeneration locally via NAD (+) construction, science,348 2016, pages 453-457, hereby incorporated by reference in its entirety.) neurodegenerative diseases and injuries are devastating to both the patient and caregivers.

Disarm Therapeutic, inc. Filed WO2019236890A1 and WO2020247701A2 disclose SARM1 inhibitor compounds having the following structure, respectively, for inhibiting SARMI and/or treating and/or preventing axonal degeneration:

disarm Therapeutic, inc. Filed WO2020081923A1 and WO2020132045A1 respectively relate to the combined use of such SARM1 inhibitor compounds with other agents.

Basheer et al (Ahmad Basheer, zvi rapport, journal of Organic Chemistry (2008), 73 (4), 1386-1396) discuss the chemical reaction of thiols derived from cyano-monothiocarbonyl malonamide and cyano-dithiocarbonyl malonamide, disclose compounds having the following structure, but do not disclose any pharmacological activity of the compounds:

WO2004011461A1 relates to isothiazole derivatives having TrkA receptor, KDR tyrosine kinase, receptor tyrosine kinase, VEGF-2 and TrkB inhibitory activity for use as anti-cancer agents. This international patent application discloses intermediate 14 for the synthesis of its subject compounds, having the following structure, but does not disclose any pharmacological activity thereof:

lippa et al (Blaise Lippa,Joel Morris,Matthew Corbett,Bioorganic&Medicinal Chemistry Letters,16(2006),3444-3448) disclose intermediate 13 for the synthesis of isothiazole inhibitors of TrkA kinase, but do not disclose any of its pharmacological activities:

There remains a need to develop new, in particular more active, inhibitors of SARM1 to meet clinical needs.

Disclosure of Invention

The inventors surprisingly found that when the oxo (= O) group on 2, 3-dihydroisothiazole of the compounds disclosed in WO2019236890A1 (e.g. positive controls 1 and 2 determined in the biological examples below) is replaced by a thio (= S), new compounds are obtained which have a significantly improved SARM1 inhibitory activity compared to the corresponding compounds. The compounds of the present disclosure also have a variety of excellent properties, such as good physicochemical properties (e.g., solubility, physical and/or chemical stability), good pharmacokinetic properties (e.g., good drug exposure and good oral absorption effects), good safety (lower toxicity and/or fewer side effects, wider therapeutic window).

Accordingly, in some embodiments, the present disclosure provides novel compounds having improved SARM1 inhibitory activity compared to the prior art for use in the treatment and/or prevention of neurodegeneration (e.g., for reducing axonal degeneration), which compounds are compounds of formula I:

Or alternatively

Wherein:

R ¹ is selected from -CN、-NO₂、-C(O)R^1a、-S(O)₂R^1a、-CONR^1aR^1b、-S(O)₂NR^1aR^1b、-C(＝NR^1a)NR^1aR^1b、-CO₂R^1a、3 to 7 membered saturated or partially unsaturated carbocyclyl, 4 to 7 membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5 or 6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and 8 to 10 membered bicyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the carbocyclyl, heterocyclyl, aryl and heteroaryl are each substituted with 0,1, 2,3 or 4R ^x;

R ² is selected from the group consisting of hydrogen, optionally substituted C _1-6 aliphatic, -C (O) R ^2a、-S(O)₂R^2a, and-CO ₂R^2a,

R ³ is- (CH ₂)_n Cy) and n is 0, 1 or 2;

Or alternatively

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

Cy is selected from the group consisting of 3-to 7-membered saturated or partially unsaturated carbocyclyl, 8-to 10-membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and 8-to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the carbocyclyl, heterocyclyl, aryl and heteroaryl are each substituted with 0, 1,2, 3 or 4R ^x;

R ⁴ is selected from the group consisting of hydrogen, optionally substituted C _1-6 aliphatic, -C (O) R ^4a、-S(O)₂R^4a、-CO₂R^4a, 3 to 7 membered saturated or partially unsaturated carbocyclyl, 4 to 7 membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5 or 6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the carbocyclyl, heterocyclyl, aryl and heteroaryl are each substituted with 0, 1,2,3 or 4R ^x; and

Each R ^x is independently selected from halogen, -CN, -NO ₂, -OH, -O-optionally substituted C _1-6 aliphatic, -SH, -S-optionally substituted C _1-6 aliphatic 、-NR^3aR^3b、-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a、-CONR^3aR^3b、-N(R^3a)SO₂R^3b、-N(R^3a)C(O)R^3b、 optionally substituted C _1-6 aliphatic, optionally substituted 5-to 6-membered saturated or partially unsaturated heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur, and optionally substituted 8-to 10-membered heteroaryl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur;

R ^1a、R^1b、R^2a、R^3a、R^3b and R ^4a are each independently hydrogen, optionally substituted C _1-6 aliphatic, optionally substituted phenyl, or optionally substituted 3-to 7-membered saturated or partially unsaturated carbocycle; or alternatively

R ^1a and R ^1b, and/or R ^3a and R ^3b, together with the nitrogen atom to which they are attached, form a 3-to 6-membered saturated or partially unsaturated heterocyclic ring;

provided that the compound is not

In some embodiments, the compound of formula I has a structure as shown in one of formulas I-a, I-a-I, I-a-ii, I-a-iii, I-a-iv, I-a-v, I-a-vi, I-a-vii, I-a-viii, I-a-ix, I-a-x, I-a-xi, I-a-xii, I-a-xiii, and I-a-xiv as described below.

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

In some embodiments, the present disclosure provides pharmaceutical compositions comprising the compounds, or enantiomers, diastereomers, racemates, stereoisomers, tautomers, geometric isomers, N-oxides, metabolites, prodrugs or pharmaceutically acceptable salts, esters, solvates, hydrates, isotopically labeled compounds or prodrugs thereof; and optionally one or more pharmaceutically acceptable carriers.

In some embodiments, the present disclosure provides the compounds and/or pharmaceutical compositions that are useful in medicine, particularly for treating neurodegeneration (e.g., for reducing axonal degeneration).

In some embodiments, provided SARM1 inhibitors reduce or inhibit the binding of SARM1 to nad+. In some embodiments, provided SARM1 inhibitors bind to SARM1 within a pocket comprising one or more catalytic residues (e.g., a catalytic cleft of SARM 1).

In some embodiments, the provided compounds and/or pharmaceutical compositions inhibit the activity of SARM 1.

Alternatively or additionally, in some embodiments, the provided compounds alleviate one or more properties of neurodegeneration.

In some embodiments, the present disclosure provides methods of treating a neurodegenerative disease, disorder, or condition associated with axonal degeneration.

In some embodiments, the compounds and/or pharmaceutical compositions described herein may be used, for example, in medical practice.

In some embodiments, the compounds and/or pharmaceutical compositions described herein are useful, for example, in the treatment, prevention, or amelioration of axonal degeneration (e.g., one or more features or characteristics thereof).

In some embodiments, the compounds and/or pharmaceutical compositions described herein are useful, for example, in inhibiting axonal degeneration, including those caused by nad+ reduction or depletion.

In some embodiments, the compounds and/or compositions described herein may be used, for example, to prevent axonal degeneration distal to an axonal injury.

In some embodiments, the compounds and/or pharmaceutical compositions described herein may be used, for example, to treat one or more neurodegenerative diseases, disorders, or conditions selected from neuropathy or axonal lesions.

In some embodiments, the compounds and/or pharmaceutical compositions described herein may be used, for example, to treat neuropathy or axonal lesions associated with axonal degeneration.

In some embodiments, the neuropathy associated with axonal degeneration is hereditary or congenital neuropathy or axonal lesions.

In some embodiments, the neuropathy associated with axonal degeneration is caused by neogenesis or somatic mutation.

In some embodiments, the neuropathy associated with axonal degeneration is selected from the list contained herein.

In some embodiments, the neuropathy or axonal lesions 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 disease, ischemia or stroke, chemical injury, thermal injury, and AIDS.

In some embodiments, an individual administered a compound or pharmaceutical composition described herein may be or include an individual suffering from or susceptible to a neurodegenerative disease, disorder, or condition.

In some embodiments, the neurodegenerative disease, disorder, or condition may be or comprise traumatic neuronal injury.

In some embodiments, the traumatic neuronal injury is a blunt trauma, closed head injury, open head injury, exposure to concussive and/or explosive forces, penetrating injury within or to a brain cavity or body innervation area.

In some embodiments, the traumatic neuronal injury is a force that causes axonal deformation, stretching, squeezing, or shearing.

In some embodiments, provided methods comprise administering a compound described herein (e.g., a compound of formula I) to a patient in need thereof.

In some such embodiments, the patient is at risk of suffering from a disease, disorder, or condition characterized by axonal degeneration.

In some embodiments, the patient has a disease, disorder, or condition characterized by axonal degeneration.

In some embodiments, the patient has been diagnosed with a disease, disorder, or condition characterized by axonal degeneration.

In some embodiments, provided methods comprise administering a pharmaceutical composition described herein to a population of patients in need thereof.

In some embodiments, the population is from those individuals involved in activities with a high likelihood of traumatic neuronal injury.

In some embodiments, the population is from those athletes participating in contact sports or other high risk activities.

In some embodiments, the patient is at risk of suffering from 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.

In some 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 are also useful for studying the function of SARM1 in biological and pathological phenomena and for comparative evaluation of novel inhibitors of SARM1 activity in vitro or in vivo.

In some embodiments, the compounds and/or pharmaceutical compositions described herein are useful as methods of, for example, inhibiting degeneration of neurons derived from an individual.

In some embodiments, the compounds and/or pharmaceutical compositions described herein may be used to inhibit the degeneration of neurons or a portion thereof in vitro culture.

In some embodiments, the compounds and/or pharmaceutical compositions described herein may be used as stabilizers to promote neuronal survival in vitro.

Brief description of the drawings

FIG. 1 shows the structure of SARM1 protein.

Definition of the definition

Aliphatic: the term "aliphatic" refers to a straight (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more units of unsaturation, having a single point of attachment to the remainder of the molecule. Unless otherwise indicated, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, the aliphatic group contains 1 to 5 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1 to 4 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1-3 aliphatic carbon atoms, and in other embodiments, the aliphatic group contains 1-2 aliphatic carbon atoms. Suitable aliphatic groups include, but are not limited to, straight or branched chain, substituted or unsubstituted alkyl, alkenyl, alkynyl, and hybrids thereof.

Cycloaliphatic: the term "cycloaliphatic" refers to a mono-or bicyclic hydrocarbon (also referred to herein as "carbocycle" or "cycloaliphatic") that is fully saturated or contains one or more units of unsaturation, but which is not aromatic, and which has a single point of attachment to the remainder of the molecule. In some embodiments, "cycloaliphatic" (or "carbocycle") refers to a C ₃-C₈ monocyclohydrocarbon or a C ₇-C₁₀ bicyclohydrocarbon that is fully saturated or contains one or more unsaturated units, but which is not aromatic, having a single point of attachment to the remainder of the molecule.

Alkyl: the term "alkyl" as used alone or as part of a larger group or moiety (mole) refers to a saturated, optionally substituted, straight or branched hydrocarbon group having 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2 carbon atoms.

Cycloalkyl: the term "cycloalkyl" as used alone or as part of a larger group or moiety (mole) refers to an optionally substituted saturated mono-or bicyclic hydrocarbon ring system having 3 to 10 ring carbon atoms. In some embodiments, "cycloalkyl" is a monocyclic C ₃-C₈ cycloalkyl. In some embodiments, "cycloalkyl" is a bicyclic C ₇-C₁₀ cycloalkyl. Exemplary monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

An alkylene group: the term "alkylene" refers to a divalent alkyl group. In some embodiments, "alkylene" is a divalent straight or branched chain alkyl group. In some embodiments, the "alkylene chain" is a polymethylene group, i.e., - (CH ₂)_n -, wherein n is a positive integer, such as 1 to 6, 1 to 4,1 to 3, 1 to 2, or 2 to 3. The optionally substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are optionally replaced with substituents suitable substituents include those described below for the substituted aliphatic group, it will be appreciated that the two substituents of the alkylene group may together form a ring system, the two substituents may together form a 3-to 7-membered ring.

Alkenyl: the term "alkenyl" used alone or as part of a larger group or moiety (mole) refers to an optionally substituted straight or branched chain 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 mono-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.

Alkenyl: the term "alkenyl" used alone or as part of a larger group or moiety (mole) refers to an optionally substituted straight or branched chain 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.

Cycloalkenyl group: the term "cycloalkenyl" as used alone or as part of a larger group or moiety refers to an optionally substituted non-aromatic mono-or multicyclic hydrocarbon ring system containing at least one carbon-carbon double bond and having 3 to 10 carbon atoms. Exemplary monocyclic cycloalkenyl groups include cyclopentenyl, cyclohexenyl, and cycloheptenyl.

Alkynyl: the term "alkynyl" used alone or as part of a larger group or moiety (mole) refers to an optionally substituted straight 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.

Aryl: the term "aryl" (or "aromatic ring") refers to an all-carbon monocyclic or fused-polycyclic aromatic group having a conjugated pi-electron system. As used herein, the term "C ₆-₁₂ aryl (aromatic ring)" means an aryl (aromatic ring) containing 6 to 12 carbon atoms, preferably a C ₆-₁₀ aryl (aromatic ring), preferably phenyl or naphthyl. Aryl is optionally substituted with one or more (such as 1to 3) identical or different substituents (e.g., halogen, OH, CN, NO ₂、C₁-C₆ alkyl, etc.).

Aryl: the term "aryl" used alone or as part of a larger group or moiety (moiety) refers to an all-carbon monocyclic or fused-polycyclic ring system having a conjugated pi-electron system. The term "aryl" may be used interchangeably with the term "aryl ring" or "aromatic ring". In some embodiments, "aryl" refers to an aryl group containing 6 to 12 ring carbon atoms (C ₆-₁₂ aryl). In some embodiments, "aryl" refers to an aryl group containing 6 to 10 ring carbon atoms (C ₆-₁₀ aryl). Aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and the like. Aryl groups may be optionally substituted.

Heteroaryl group: the term "heteroaryl" used alone or as part of a larger group or moiety (e.g., "heteroaralkyl" or "heteroarylalkoxy") refers to the following group: having 5 to 10 ring atoms, preferably 5, 6, 8, 9 or 10 ring atoms; having 6, 10 or 14 pi electrons shared in a circular array; in addition to carbon atoms, there are 1 to 5 (e.g., 1-4, 1-3, or 1-2) heteroatoms. The term "heteroatom" refers to nitrogen, oxygen or sulfur, and includes any oxidized form of nitrogen or sulfur as well as any quaternized form of basic nitrogen. Heteroaryl groups may be optionally substituted. Heteroaryl groups include, but are 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 term "heteroaryl" as used herein also includes groups in which a heteroaromatic ring (e.g., a 5 or 6 membered heteroaryl ring) is fused to one or more aryl rings (e.g., benzene rings), wherein the linking group or point of attachment is on the heteroaromatic ring. In some embodiments of such heteroaryl groups, the heteroatoms are all in the heteroaromatic ring (e.g., 5 or 6 membered heteroaryl ring) and are not common atoms. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, oxazinyl. Heteroaryl groups may be monocyclic or bicyclic. The term "heteroaryl (heteroaryl)" may be used interchangeably with the terms "heteroaryl ring", "heteroaryl group" or "heteroaromatic", any of which include optionally substituted rings. The term "heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl moieties are independently optionally substituted.

Heterocycles: as used herein, the terms "heterocycle (heterocycle)", "heterocyclyl (heterocyclyl)", "heterocyclyl (heterocyclic radical)", and "heterocycle (heterocyclic ring)" are used interchangeably to include stable 3-to 8-membered (e.g., 4,5,6, or 7-membered) monocyclic or 7-10 (e.g., 8, 9, or 10-membered) bicyclic heterocyclic moieties that are saturated or partially unsaturated, having one or more, such as 1 to4 (e.g., 1-3, 1-2, or 2-3) heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen or sulfur, and includes any oxidized form of nitrogen or sulfur as well as any quaternized form of basic nitrogen. The term "nitrogen" when used in reference to a ring atom of a heterocycle includes substituted nitrogen. For example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur and nitrogen, the nitrogen may be N (as in 3, 4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR ⁺ (as in N-substituted pyrrolidinyl). In some embodiments, the term "heterocycle" is a 5 or 6 membered saturated or partially unsaturated monocyclic heterocyclyl having 1,2, or 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the term "heterocycle" is an 8-to 10-membered saturated or partially unsaturated bicyclic heterocyclyl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. The heterocycle may be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any ring atom may be optionally substituted. Examples of such saturated or partially unsaturated heterocyclyl groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepine (diazepinyl), oxazepinyl (oxazepinyl), thiazepinyl (thiazepinyl), morpholinyl, and thiomorpholinyl. The term "heterocycle" also includes within its scope a bicyclic ring system wherein the benzene ring is fused to a non-aromatic heterocycle, and a bicyclic ring system wherein the heteroaryl group is fused to a non-aromatic heterocycle or cycloaliphatic ring as defined above, wherein any of the bicyclic ring systems is non-aromatic as a whole, and wherein the linking group or point of attachment is on the benzene ring or heteroaryl ring. In some such embodiments, such bicyclic heterocyclyl is an 8-to 10-membered (particularly 9-or 10-membered) bicyclic heterocyclyl having 1-3 (e.g., 1,2, or 3) heteroatoms selected independently from nitrogen, oxygen, and sulfur. In some such embodiments, such bicyclic heterocyclyl is a fused ring system of a benzene ring with a 5 or 6 membered saturated or partially unsaturated heterocyclyl ring, wherein the heteroatoms are all in the 5 or 6 membered saturated or partially unsaturated heterocyclyl ring and are not common atoms. Examples of such bicyclic heterocyclic groups include, but are not limited to, 2, 3-dihydrobenzofuranyl, 2, 3-dihydrobenzothienyl, indolinyl, 2, 3-dihydrobenzoindazolyl, 2, 3-dihydrobenzimidazolyl, chromanyl, thiochroman, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido [2,3-b ] -1, 4-oxazin-3 (4H) -one. Heterocyclyl groups may also include tricyclic or polycyclic ring systems. Preferably a monocyclic, bicyclic or tricyclic heterocyclyl, more preferably a monocyclic or bicyclic heterocyclyl, as defined above. The term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group, wherein the alkyl and heterocyclyl moieties are independently optionally substituted.

"Substituted" or "optionally substituted": as described herein, the compounds of the present invention may contain "optionally substituted" groups or moieties. In general, the term "optionally substituted" means that one or more hydrogens of the designated group or moiety may or may not be replaced with a suitable substituent. "substituted" applies to one or more hydrogens either explicitly or implicitly in the structure (e.g.,At least refer toWhileAt least refer to ). Unless otherwise indicated, an "optionally substituted" group may have suitable substituents at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from the specified group, the substituents may be the same or different at each position. The combinations of substituents envisaged by the present invention are preferably those which lead to the formation of stable or chemically viable compounds. The term "stable" as used herein means that the compound does not substantially change when subjected to conditions that allow for its production, detection, and in certain embodiments, its recovery, purification, and use for one or more of the purposes disclosed herein.

Suitable monovalent substituents on the substitutable carbon or nitrogen atom of the "optionally substituted" group are independently halogen ;-(CH₂)_0-4R^○;-(CH₂)_0-4OR^○;-O(CH₂)_0-4R^○;-O-(CH₂)_0-4C(O)OR^○;-(CH₂)_0-4CH(OR^○)₂;-(CH₂)_0-4SR^○;-(CH₂)_0-4Ph, which may be substituted with R ^○; - (CH ₂)_0-4O(CH₂)_0-1 Ph, which may be substituted by R ^○; -ch=chph, which may be substituted with R ^○; - (CH ₂)_0-4O(CH₂)_0-1 -pyridinyl, which may be substituted by R ^○ for ;-NO₂;-CN;-N₃;-(CH₂)_0-4N(R^○)₂;-(CH₂)_0-4N(R^○)C(O)R^○;-N(R^○)C(S)R^○;-(CH₂)_0-4N(R^○)C(O)NR^○ ₂;-N(R^○)C(S)NR^○ ₂;-(CH₂)_0-4N(R^○)C(O)OR^○;-N(R^○)N(R^○)C(O)R^○;-N(R^○)N(R^○)C(O)NR^○ ₂;-N(R^○)N(R^○)C(O)OR^○;-(CH₂)_0-4C(O)R^○;-C(S)R^○;-(CH₂)_0-4C(O)OR^○;-(CH₂)_0-4C(O)SR^○;-(CH₂)_0-4C(O)OSiR^○ ₃;-(CH₂)_0-4OC(O)R^○;-OC(O)(CH₂)_0-4SR^○;-(CH₂)_0-4SC(O)R^○;-(CH₂)_0- ₄C(O)NR^○ ₂;-C(S)NR^○ ₂;-C(S)SR^○;-SC(S)SR^○;-(CH₂)_0-4OC(O)NR^○ ₂;-C(O)N(OR^○)R^○;-C(O)C(O)R^○;-C(O)CH₂C(O)R^○;-C(NOR^○)R^○;-(CH₂)_0-4SSR^○;-(CH₂)_0-4S(O)₂R^○;-(CH₂)_0- ₄S(O)₂OR^○;-(CH₂)_0-4OS(O)₂R^○;-S(O)₂NR^○ ₂;-(CH₂)_0-4S(O)R^○;-N(R^○)S(O)₂NR^○ ₂;-N(R^○)S(O)₂R^○;-N(OR^○)R^○;-C(NH)NR^○ ₂;-P(O)₂R^○;-P(O)R^○ ₂;-OP(O)R^○ ₂;-OP(O)(OR^○)₂;-SiR^○ ₃;-(C_1-4 linear or branched alkylene) O-N (R ^○)₂; Or- (C _1-4 linear or branched alkylene) C (O) O-N (R ^○)₂, wherein each R ^○ may be substituted as defined below and is independently hydrogen, C _1-6 aliphatic, -CH ₂Ph、-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, in spite of the above definition, two independently occurring R ^○ together with their intermediate atoms form a ring having 0-4 heteroatoms independently selected from nitrogen, A3 to 12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic or bridged ring of heteroatoms of oxygen or sulfur, which may be substituted as defined below. in some embodiments, suitable monovalent substituents on the substitutable carbon or nitrogen atom of an "optionally substituted" group are selected from halogen, -NO ₂、-CN、OH、SH、C_1-6 alkyl, -O-C _1-6 alkyl, -C (O) OC _1-6 alkyl, -NH ₂、-NH(C_1-6 alkyl) and-N (C _1-6 alkyl) ₂.

Suitable monovalent substituents on R ^○ (OR the two independently occurring R ^○ together with the ring they form together with the intervening atom) are independently halogen, - (CH ₂)_0-2R^·, - (halo R^·)、-(CH₂)_0-2OH、-(CH₂)_0-2OR^·、-(CH₂)_0-2CH(OR^·)₂、-O( halo R^·)、-CN、-N₃、-(CH₂)_0-2C(O)R^·、-(CH₂)_0-2C(O)OH、-(CH₂)_0-2C(O)OR^·、-(CH₂)_0- ₂SR^·、-(CH₂)_0-2SH、-(CH₂)_0-2NH₂、-(CH₂)_0-2NHR^·、-(CH₂)_0-2NR^· 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 substituted with only one OR more halogen in the case of the preceding "halo", and are independently selected from C _1-4 aliphatic, -CH ₂Ph、-O(CH₂)_0-1 Ph OR 3 to 6 membered saturated, partially unsaturated OR aryl rings having 0 to 4 heteroatoms independently selected from nitrogen, oxygen OR sulfur suitable divalent substituents on the saturated carbon atoms of R ^○ include =o and =s.

Suitable divalent substituents on the saturated carbon atoms of the "optionally substituted" group include the following: =o ("oxo ")、＝S、＝NNR^* ₂、＝NNHC(O)R^*、＝NNHC(O)OR^*、＝NNHS(O)₂R^*、＝NR^*、＝NOR^*、-O(C(R^* ₂))_2-3O- or-S (C (R ^* ₂))_2-3 S-, wherein each independently occurring R ^* is selected from hydrogen, a substituted C _1-6 aliphatic, as may be 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 attached to the vicinal substitutable carbon of the" optionally substituted "group include-O (CR ^* ₂)_2-3 O-, wherein each independently occurring R ^* is selected from hydrogen, a substituted C _1-6 aliphatic, as may be 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 substituents on the aliphatic group of R ^* include halogen, -R ^·, - (halo R ^·)、-OH、-OR^·, -O (halo R ^·)、-CN、-C(O)OH、-C(O)OR^·、-NH₂、-NHR^·、-NR^· ₂ or-NO ₂, wherein each R ^· is unsubstituted, or in the case of a "halo" preceding it is substituted with one or more halogens only, and is independently C _1-4 aliphatic, -CH ₂Ph、-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.

Suitable substituents on the substitutable nitrogen of an "optionally substituted" group include those that include Each of which is provided withIndependently hydrogen, a substituted C _1-6 aliphatic, unsubstituted-OPh, which may be defined as hereinafter, 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 two independently occurring in spite of the above definitionTogether with their intermediate atoms, form an unsubstituted 3 to 12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

At the position ofSuitable substituents on the aliphatic groups of (a) are independently halogen, -R ^·, - (halo R ^·)、-OH、-OR^·, -O (halo R ^·)、-CN、-C(O)OH、-C(O)OR^·、-NH₂、-NHR^·、-NR^· ₂ or-NO ₂, wherein each R ^· is unsubstituted or substituted with only one or more halogens in the case of a preceding "halo"), and are independently C _1-4 aliphatic, -CH ₂Ph、-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.

Halogen: as used herein, the term "halogen" or "halo" group is defined to include fluorine, chlorine, bromine or iodine.

Partially unsaturated: as used herein, the term "partially unsaturated" refers to a cyclic group or 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.

Combining: it is to be understood that the term "binding" as used herein generally refers to non-covalent association between or among two or more entities. "direct" bonding involves physical contact between entities or parts; indirect bonding involves physical interaction by means of physical contact with one or more intermediate entities. Binding between two or more entities can generally be assessed in any of a variety of situations, including studying the interacting entities or portions 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).

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., a 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 may be or comprise bone marrow; blood; blood cells; ascites; tissue or fine needle biopsy samples; a body fluid comprising cells; free nucleic acid; sputum; saliva; urine; cerebrospinal fluid; peritoneal fluid, pleural fluid; feces; lymph; gynecological liquid; a skin swab; a vaginal swab; an oral swab; a nasal swab; wash or lavage, such as catheter lavage or bronchoalveolar lavage; liquid suction; a wiper blade; bone marrow specimens; tissue biopsy specimens; surgical specimens; feces, other body fluids, secretions and/or excretions; and/or cells obtained therefrom, and the like. 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 an individual from whom the sample was obtained. In some embodiments, the sample is a "raw sample" obtained directly from a source of interest by any suitable means. For example, in some embodiments, the original biological sample is obtained by a method selected from biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of bodily fluids (e.g., blood, lymph, stool, etc.), and the like. In some embodiments, as will be appreciated from the context, the term "sample" refers to a preparation obtained by processing a raw sample (e.g., by removing one or more components of the raw sample and/or by adding one or more reagents thereto). For example, filtration is performed using a semipermeable membrane. Such "treated samples" may comprise, for example, nucleic acids or proteins extracted from the sample or obtained by subjecting the original sample to techniques such as amplification or reverse transcription of mRNA, isolation and/or purification of certain components, and the like.

Biomarkers: the term "biomarker" is used herein to refer to an entity, event, or feature whose presence, level, extent, type, and/or form is associated with a particular biological event or state of interest, and thus is considered a "marker" for that event or state. In some embodiments, the biomarker may be or comprise a marker of a particular disease state, or a marker of the likelihood that a particular disease, disorder, or condition may develop, or relapse, to name a few. In some embodiments, the biomarker may be or comprise a marker of a particular disease or its therapeutic outcome or likelihood. Thus, in some embodiments, the biomarker is a prediction of a related biological event or state of interest, in some embodiments, the biomarker is a prognosis of a related biological event or state of interest, in some embodiments, the biomarker is a diagnosis of a related biological event or state of interest. The biomarker may be or comprise an entity of any chemical class, and may be or comprise a combination of entities. For example, in some embodiments, the biomarker may be or comprise a nucleic acid, polypeptide, lipid, carbohydrate, small molecule, inorganic agent (e.g., metal or ion), or a combination thereof. In some embodiments, the biomarker is a cell surface marker. In some embodiments, the biomarker is intracellular. In some embodiments, the biomarker is detected extracellularly (e.g., is secreted or otherwise produced or present extracellularly, e.g., in bodily fluids such as blood, urine, tears, saliva, cerebrospinal fluid, etc. in some embodiments, the biomarker may be or comprise a genetic or epigenetic feature.

In some embodiments, the biomarker may be or comprise a marker of neurodegeneration, or a marker of likelihood that a neurodegenerative disease, disorder, or condition may develop, or relapse. In some embodiments, the biomarker may be or comprise a marker of neurodegeneration, its therapeutic outcome, or likelihood. Thus, in some embodiments, the biomarker is a prognosis of a neurodegenerative disease, disorder, or condition, in some embodiments, the biomarker is a diagnosis of a neurodegenerative disease, disorder, or condition. In some embodiments, the change in biomarker levels may be detected by cerebrospinal fluid (CSF), plasma, and/or serum.

In some embodiments, neurodegeneration may be assessed, 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 cerebrospinal fluid of an individual. In some embodiments, the occurrence and/or progression of neurodegeneration can be assessed by Positron Emission Tomography (PET) with a synaptobrevin 2A (SV 2A) ligand. In some embodiments, detectable changes in constitutive NAD and/or cADPR levels in neurons can be used to assess neurodegeneration.

In some embodiments, a detectable change in one or more neurodegeneration associated proteins in an individual relative to a healthy reference population can be used as a biomarker for neurodegeneration. Such proteins include, but are not limited to, albumin, amyloid-beta (aβ) 38, aβ40, aβ42, glial Fibrillary Acidic Protein (GFAP), cardiac fatty acid binding protein (hFABP), monocyte Chemotactic Protein (MCP) -1, neuroparticulate protein, neuron-specific enolase (NSE), soluble amyloid precursor protein (sAPP) alpha, sappβ, soluble trigger receptor 2 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, may be used as biomarkers of neurodegeneration.

And (3) a carrier: as used herein, the term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the (pharmaceutical) composition is administered. In some exemplary embodiments, the carrier may include sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. In some embodiments, the carrier is or includes one or more solid components.

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

Composition: those skilled in the art will appreciate that the term "composition" may be used to refer to a discrete physical entity comprising one or more specified components. In general, unless otherwise specified, the composition may be in any form, such as a gas, gel, liquid, solid, and the like.

Domain: the term "domain" as used herein refers to a section or portion of an entity. In some embodiments, a "domain" is associated with a particular structural and/or functional feature of an entity, such that when the domain is physically separated from the remainder of its parent entity, it substantially or entirely retains the particular structural and/or functional feature. Alternatively or additionally, a domain may be or comprise a part of an entity which, when separated from the (parent) entity and connected to a different (acceptor) entity, substantially retains and/or confers to the acceptor entity one or more structural and/or functional features which are characteristic in the parent entity. In some embodiments, a domain is a segment or portion of a molecule (e.g., a small molecule, a carbohydrate, a lipid, a nucleic acid, or a polypeptide). In some embodiments, the domain is a segment of a polypeptide; in some such embodiments, the domain is characterized by a particular structural element (e.g., a particular amino acid sequence or sequence motif, an alpha-helical feature, a beta-sheet feature, a coiled-coil feature, a random coiled-coil feature, etc.), and/or by a particular functional feature (e.g., binding activity, enzymatic activity, folding activity, signaling activity, etc.).

Dosage form or unit dosage form: those skilled in the art will appreciate that the term "dosage form" may be used to refer to physically discrete units of an active agent (e.g., a therapeutic agent or diagnostic agent) for administration to an individual. Typically, each such unit contains a predetermined amount of active agent. In some embodiments, such amounts are unit doses (or all fractions thereof) suitable for administration in accordance with a dosing regimen (i.e., a therapeutic dosing regimen) that has been determined to be relevant to the desired or beneficial outcome when administered to the relevant population. Those of ordinary skill in the art will appreciate that the total amount of therapeutic composition or agent administered to a particular individual is determined by one or more attending physicians and may involve the administration of multiple dosage forms.

Dosing regimen or treatment regimen: those skilled in the art will appreciate that the terms "dosing regimen" and "treatment regimen" may be used to refer to a set of unit doses (typically more than one) administered individually to an individual, which unit doses are typically 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 comprises a plurality of doses, wherein each dose is separated in time from the other doses. In some embodiments, the individual doses are spaced from each other by a period of the same length; in some embodiments, the dosing regimen includes a plurality of doses and at least two different time periods separating the individual doses. In some embodiments, all doses within a dosing regimen have the same amount of unit dose. In some embodiments, different doses within a dosing regimen have different amounts. In some embodiments, the dosing regimen comprises a first dose in an amount of the first dose followed by one or more additional doses in an amount of the second dose that is different from the amount of the first dose. In some embodiments, the dosing regimen comprises a first dose in an amount of the first dose followed by one or more additional doses in an amount of the second dose that is the same as the amount of the first dose. In some embodiments, the dosing regimen is associated with a desired or beneficial outcome (i.e., is a therapeutic dosing regimen) when administered in the relevant population.

Excipient: as used herein, the term "excipient" refers to a non-therapeutic agent that may be included in a pharmaceutical composition, e.g., to provide or promote a desired consistency or stabilization. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like.

Inhibitors: as used herein, the term "inhibitor" refers to an entity, condition, or event whose presence, level, or extent correlates to a decrease in the level or activity of a target. In some embodiments, the inhibitor may act directly (in which case it directly exerts an effect on its target, e.g., by binding to the target); in some embodiments, the inhibitor may act indirectly (in which case it exerts an effect by interacting with and/or otherwise altering a modulator of the target, causing a decrease in the level and/or activity of the target). In some embodiments, an inhibitor is one whose presence or level is correlated with a reduced target level or activity relative to a particular reference level or activity (e.g., a level or activity observed in the presence of a known inhibitor, or in the absence of the inhibitor in question, etc., under appropriate reference conditions).

Neurodegeneration: as used herein, the term "neurodegeneration" refers to a decrease in one or more characteristics, structures, or properties of a neuron or neuronal tissue. In some embodiments, neurodegeneration is observed as a pathological decrease in an organism. Those of skill in the art will appreciate that neurodegeneration is associated with certain diseases, disorders, and conditions, including those affecting humans. In some embodiments, neurodegeneration may be transient (e.g., sometimes occurring in association with certain infections and/or chemical or mechanical destruction); in some embodiments, the neurodegeneration may 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's disease (Huntington disease), or alzheimer's disease). In some embodiments, neurodegeneration can be assessed, for example, by detecting an increase in a biomarker associated with neurodegeneration in an individual. In some embodiments, neurodegeneration can be assessed, for example, by detecting a decrease in a biomarker associated with neurodegeneration in an individual. Alternatively or additionally, in some embodiments, neurodegeneration may be assessed by Magnetic Resonance Imaging (MRI), cerebrospinal fluid containing biomarkers, or other biomarkers observed in the patient. In some embodiments, neurodegeneration is defined as a score of less than 24 for a small mental state examination. In some embodiments, neurodegeneration refers to loss of synapses. In some embodiments, neurodegeneration refers to a decrease in nerve tissue associated with traumatic injury (e.g., exposure to external forces that disrupt the integrity of the nerve tissue). In some embodiments, neurodegeneration refers to a decrease in peripheral nerve tissue. In some embodiments, neurodegeneration refers to a decrease in central nervous tissue.

Oral administration: the phrases "oral administration (oral administration)" and "oral administration (ADMINISTERED ORALLY)" as used herein have their art-understood meanings, and refer to administration of a compound or composition by the oral cavity.

Parenteral: the phrases "parenteral administration (PARENTERAL ADMINISTRATION)" and "parenteral administration (ADMINISTERED PARENTERALLY)" as used herein have their art-understood meaning to refer to modes of administration other than enteral and topical administration (typically by injection), and include, but are not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

Patient: as used herein, the term "patient" refers to any organism to which the provided composition is or may be administered, e.g., 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 is suffering from or susceptible to one or more disorders or conditions. In some embodiments, the patient exhibits one or more symptoms of the disorder or condition. In some embodiments, the patient has been diagnosed as having one or more disorders or conditions. In some embodiments, the patient is receiving or has received certain therapies to diagnose and/or treat a disease, disorder, or condition.

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 treatment or dosing regimen that, when administered to a relevant population, exhibits a statistically significant probability of achieving a predetermined therapeutic effect. In some embodiments, the pharmaceutical composition may be specifically formulated for administration in solid or liquid form, including pharmaceutical compositions suitable for: oral administration, e.g., drenching (aqueous or nonaqueous solutions or suspensions), tablets, e.g., tablets, boluses, powders, granules, pastes for application to the tongue, targeted for intra-buccal, sublingual, and systemic absorption; parenteral administration, for example, as a sterile solution or suspension or sustained release formulation, by subcutaneous, intramuscular, intravenous or epidural injection; topical application, for example as a cream, ointment or controlled release patch or spray to the skin, lungs or oral cavity; intravaginal or intrarectal, for example as pessaries, creams or foams; sublingual; ocular menstruation; percutaneous; or transnasal, pulmonary and other mucosal surfaces.

Pharmaceutically acceptable: as used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions, and/or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

A pharmaceutically acceptable carrier: as used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, component, 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 body part to another organ or body part. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the patient. Some examples of materials that may 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; powdery 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; diols such as propylene glycol; polyols such as glycerol, 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 physiological saline; ringer's solution; ethanol; a pH buffer solution; polyesters, polycarbonates and/or polyanhydrides; and other non-toxic compatible substances employed in pharmaceutical formulations.

A pharmaceutically acceptable salt: the term "pharmaceutically acceptable salt" as used herein refers to salts of such compounds which are suitable for use in a pharmaceutical setting, i.e., salts which are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit/risk ratio, within the scope of sound medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, S.M. Berge et al, J.pharmaceutical Sciences, 66:1-19 (1977) describe pharmaceutically acceptable salts in detail. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts, which are salts of amino groups with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or salts formed by using other methods used in the art, such as ion exchange methods. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, adipates, alginates, ascorbates, aspartate, benzenesulfonates, benzoates, bisulphates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecyl sulfates, ethane sulfonates, formates, fumarates, glucoheptonates, glycerophosphate, gluconate, hemisulfate, heptanoates, caprates, hydroiodides, 2-hydroxy-ethane sulfonates, lactonates, lactates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmates, pamonates, pectinates, persulfates, 3-phenylpropionates, phosphates, bitrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. In some embodiments, pharmaceutically acceptable salts include nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl groups having 1 to 6 carbon atoms, sulfonate, and arylsulfonate, as appropriate.

Stereoisomers of: the term "stereoisomer" refers to an isomer formed as a result of at least one asymmetric center. In compounds having one or more (e.g., 1, 2,3, or 4) asymmetric centers, they can produce racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. Specific individual molecules may also exist as geometric isomers (cis/trans). Similarly, the compounds provided by the present disclosure may exist as a mixture of two or more structurally distinct forms (commonly referred to as tautomers) in rapid equilibrium. Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, and the like. It is to be understood that the scope of the present application encompasses all such isomers in any ratio (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%) or mixtures thereof.

"Diastereoisomers" refers to stereoisomers which have two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties, and reactivity. Mixtures of diastereomers can be separated by high resolution analytical methods such as electrophoresis and chromatography.

"Enantiomer" refers to two stereoisomers of a compound that are non-superimposable mirror images of each other.

The term "chiral" refers to molecules that have non-superimposability of a mirror image pair, while the term "achiral" refers to molecules that may be superimposed on their mirror image pair.

The compounds of the invention may be prepared in racemic form, or individual enantiomers may be prepared by enantioselective synthesis or by resolution.

As used herein, the term "cis-trans isomer" or "geometric isomer" is caused by the inability of a double bond or a single bond of a ring-forming carbon atom to rotate freely. The compounds provided herein include all cis, trans, cis (syn), trans (anti), entgegen (E) and zusammen (Z) isomers, and corresponding mixtures thereof.

It will also be appreciated that certain compounds of the invention may exist in free form for use in therapy or, where appropriate, in the form of pharmaceutically acceptable derivatives thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, metabolites or prodrugs which, upon administration to a patient in need thereof, are capable of providing the compounds of the present invention or metabolites or residues thereof, either directly or indirectly. Thus, when reference is made herein to "a compound of the invention" it is also intended to encompass the various derivative forms of the compounds described above.

Solid lines (-), solid wedges may be used hereinOr virtual wedge shapeDepicting the chemical bonds of the compounds provided by the present disclosure. The use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers at that carbon atom (e.g., a particular enantiomer, a racemic mixture, etc.) are included. The use of a real or virtual wedge to depict a bond to an asymmetric carbon atom is intended to indicate the presence of the stereoisomers shown. When present in a racemic mixture, real and imaginary wedges are used to define the relative stereochemistry, not the absolute stereochemistry. Unless otherwise indicated, the compounds provided by the present disclosure are intended to exist as stereoisomers (which include cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotamers, conformational isomers, atropisomers, and mixtures thereof). The compounds provided by the present disclosure may exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

Polymorphs: the present disclosure encompasses all possible crystalline forms or polymorphs of the compounds provided by the present disclosure, which may be single polymorphs or mixtures of any ratio of more than one polymorph.

A solvate, metabolite or prodrug: it will also be appreciated that certain compounds provided by the present disclosure may exist in free form for use in therapy, or in the form of pharmaceutically acceptable derivatives thereof, as appropriate. In the present disclosure, pharmaceutically acceptable derivatives include, but are not limited to: pharmaceutically acceptable salts, solvates, metabolites, or prodrugs thereof, which upon administration to a patient in need thereof, are capable of providing the compounds provided herein, or metabolites or residues thereof, directly or indirectly. Thus, when reference is made herein to "a compound provided by the present disclosure," it is also intended to encompass the various derivative forms of the compound described above.

The compounds provided herein may exist in the form of solvates (preferably hydrates) wherein the compounds provided herein comprise a polar solvent as a structural element of the compound lattice, such as, inter alia, water, methanol or ethanol. The polar solvent, in particular water, may be present in stoichiometric or non-stoichiometric amounts.

N-oxide: those skilled in the art will appreciate that not all nitrogen-containing heterocycles are capable of forming N-oxides, as nitrogen requires available lone pairs to oxidize to oxides; those skilled in the art will recognize nitrogen-containing heterocycles capable of forming N-oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include oxidizing heterocycles and tertiary amines with peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as t-butyl hydroperoxide, sodium perborate, and dioxiranes (dioxirane) such as dimethyl dioxirane. These methods for preparing N-oxides have been widely described and reviewed in the literature, see for example: T.L. Gilchrist, comprehensive Organic Synthesis, vol.7, pp 748-750; katritzky and a.j. Boulton, eds., ACADEMIC PRESS; and g.w.h.cheeseman and e.s.g.werstiuk, ADVANCES IN Heterocyclic Chemistry, vol.22, pp 390-392, a.r.katritzky and a.j.boulton, eds., ACADEMIC PRESS.

Also included within the scope of the present disclosure are metabolites of the compounds provided by the present disclosure, i.e., substances that form in vivo upon administration of the compounds provided by the present disclosure. Such products may result from, for example, oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc. of the compound being administered. Accordingly, the present disclosure includes metabolites of the compounds provided by the present disclosure, including compounds made by a method of contacting a compound provided by the present disclosure with a mammal for a time sufficient to produce a metabolite thereof.

The present disclosure further includes within its scope prodrugs of the compounds provided herein, which are certain derivatives of the compounds provided herein that may themselves have little or no pharmacological activity, which, when administered into or onto the body, may be converted to the compounds provided herein having the desired activity by, for example, hydrolytic cleavage. Typically such prodrugs will be functional derivatives of the compounds which are readily convertible in vivo into the desired therapeutically active compound. Additional information regarding the use of prodrugs can be found in "Pro-drugs as Novel DELIVERY SYSTEMS", vol.14, ACS Symposium Series (T. Higuchi and V. Stilla) and "Bioreversible CARRIERS IN Drug Design," Pergamon Press,1987 (E. B. Roche eds., american Pharmaceutical Association). Prodrugs provided by the present disclosure may be prepared, for example, by replacing appropriate functional groups present in compounds provided by the present disclosure with certain moieties known to those skilled in the art as "pro-moieties" (e.g., "Design of Prodrugs", described in h.bundwaard (Elsevier, 1985) ".

The invention also includes all pharmaceutically acceptable isotopically-labelled compounds which are identical to those of the present invention except that one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevailing in nature. Examples of isotopes suitable for inclusion in compounds of the invention include, but are not limited to, isotopes of hydrogen (e.g., deuterium (D, 2H), tritium (T, 3H)); isotopes of carbon (e.g., 11C, 13C, and 14C); isotopes of chlorine (e.g., 36 Cl); isotopes of fluorine (e.g., 18F); isotopes of iodine (e.g., 123I and 125I); isotopes of nitrogen (e.g., 13N and 15N); isotopes of oxygen (e.g., 15O, 17O, and 18O); isotopes of phosphorus (e.g., 32P); and isotopes of sulfur (e.g., 35S). Certain isotopically-labeled compounds of the present invention (e.g., those into which a radioisotope is incorporated) are useful in pharmaceutical and/or substrate tissue distribution studies (e.g., assays). The radioisotope tritium (i.e., 3H) and carbon-14 (i.e., 14C) are particularly useful for this purpose because of their ease of incorporation and ease of detection. Substitution with positron emitting isotopes (e.g., 11C, 18F, 15O, and 13N) can be used in Positron Emission Tomography (PET) studies to examine substrate receptor occupancy. Isotopically-labeled compounds of the present invention can be prepared by processes analogous to those described in the accompanying schemes and/or in the examples and preparations by substituting an appropriate isotopically-labeled reagent for the non-labeled reagent previously employed. Pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent may be isotopically substituted, e.g., D2O, acetone-D6 or DMSO-D6. In some embodiments, the isotopically-labeled compounds of the present invention are deuterated.

As used herein, the term "ester" means an ester derived from each of the compounds of the general formula in the present application, including physiologically hydrolyzable esters (compounds of the present application that can be hydrolyzed under physiological conditions to release the free acid or alcohol form). The compounds of the application may themselves be esters.

The present invention encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs or mixtures of any ratio of more than one polymorphs.

The present disclosure also encompasses compounds provided by the present disclosure that contain a protecting group. During any process of preparing the compounds provided by the present disclosure, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules of interest, thereby forming a chemically protected form of the compounds provided by the present disclosure. This can be achieved by conventional protecting groups, for example, at Protective Groups in Organic Chemistry, ed.J.F.W.McOmie, plenum Press,1973; and those described in T.W.Greene & P.G.M.Wuts, protective Groups in Organic Synthesis, john Wiley & Sons,1991, which are incorporated herein by reference. The protecting group may be removed at a suitable subsequent stage using methods known in the art.

The term "about" means within + -10%, preferably within + -5%, more preferably within + -2% of the stated value.

Prevention (Prevent) or prophylaxis (preservation): as used herein, the term "prevent" or "prevention" when used in connection with the occurrence of a disease, disorder, and/or condition refers to reducing the risk of developing the disease, disorder, and/or condition and/or delaying the onset of one or more features or symptoms of the disease, disorder, or condition. Prevention may be considered complete when the onset of a disease, disorder or condition has been delayed for a predetermined period of time.

Specificity: the term "specific", when used herein in reference to an agent having activity, is understood by those skilled in the art to mean that the agent distinguishes between potential target entities or states. For example, in some embodiments, an agent is said to "specifically" bind to one or more competing surrogate targets if the agent preferentially binds to its target in the presence of the target. In many embodiments, the specific interaction is dependent on the presence of a particular structural feature (e.g., epitope, cleft, binding site) of the target entity. It should be understood that specificity need not be absolute. In some embodiments, specificity may be assessed relative to the specificity of the binding agent for one or more other potential target entities (e.g., competitors). In some embodiments, the specificity is assessed relative to the specificity of a reference specific binding agent. In some embodiments, the specificity is assessed relative to the specificity of a reference non-specific binding agent. In some embodiments, the agent or entity does not detect binding to the competitive surrogate target under conditions that bind to its target entity. In some embodiments, the binding agent binds to its target entity with a higher binding rate, a lower dissociation rate, increased affinity, reduced dissociation, and/or increased stability as compared to the competing surrogate target.

Individuals: as used herein, the term "individual" refers to an organism, typically a mammal (e.g., a human, including in some embodiments, prenatal forms). In some embodiments, the individual has a related disease, disorder, or condition. In some embodiments, the individual is susceptible to a disease, disorder, or condition. In some embodiments, the individual exhibits one or more symptoms or features of a disease, disorder, or condition. In some embodiments, the individual does not exhibit any symptoms or features of the disease, disorder, or condition. In some embodiments, the individual is a human having one or more characteristics of a susceptibility or risk of a disease, disorder, or condition. In some embodiments, the individual is a patient. In some embodiments, the individual is an individual who is and/or has been diagnosed and/or treated.

Therapeutic agent: as used herein, the phrase "therapeutic agent" generally refers to any agent that, when administered to an organism, causes a desired pharmacological effect. In some embodiments, an agent is considered a therapeutic agent if the agent exhibits a statistically significant effect in an appropriate population. In some embodiments, the suitable population may be a population of model organisms. In some embodiments, the appropriate population may be defined by various criteria, such as a certain age group, gender, genetic background, pre-existing clinical conditions, and the like. In some embodiments, a therapeutic agent is a substance that is useful for alleviating, ameliorating, alleviating, inhibiting, preventing, delaying the onset of, reducing the severity of, and/or reducing the incidence of one or more symptoms or features of a disease, disorder, and/or condition. In some embodiments, a "therapeutic agent" refers to an agent that has been or needs to be approved by a government agency for sale to human administration. In some embodiments, a "therapeutic agent" is an agent that requires a medical prescription for administration to a human.

Treatment: as used herein, the term "treatment" or "treatment" refers to any method for partially or completely alleviating, ameliorating, alleviating, inhibiting, preventing, delaying the onset of, reducing the severity of, and/or reducing the incidence of one or more symptoms or features of a disease, disorder, and/or condition. Treatment may be administered to an individual that does not exhibit signs of a disease, disorder, and/or condition. In some embodiments, the treatment may be administered to an individual that exhibits only early signs of the disease, disorder, and/or condition, e.g., for the purpose of reducing the risk of developing a pathology associated with the disease, disorder, and/or condition.

Detailed Description

Programmed axonal degeneration and SARM1

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 by an intrinsic self-destructive process, which differs from traditional cell death pathways such as apoptosis known as Wale's degeneration (WALLERIAN DEGENERATION) (Gerdts, J. Et al, neuron,2016,89,449-460; whitmore, A.V. et al, CELL DEATH Differ, 2003,10,260-261). In the Wallace degeneration, the peripheral nerve selectively breaks down at the damaged distal axon segment, while the proximal axon segment and cell body remain intact. This denaturation is characterized by first depletion of nicotinamide mononucleotide adenyltransferase (NMNAT), followed by nicotinamide adenine dinucleotide (nad+) depletion, adenosine Triphosphate (ATP) depletion, neurofilament proteolysis, and finally axonal degeneration occurs about 8 to 24 hours after injury (Gerdts, j. Et al, neuron,2016,89,449-460).

NAD+ is a ubiquitous metabolite that plays a critical 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). Steady state regulation of nad+ levels is also responsible for maintaining stability and integrity of axons. Thus, manipulation to increase NMNAT1 axonal localization confers axonal protection (Babetto et al, cell Rep.,2010,3,1422-1429; sasaki et al, J.Neurosci., 2009).

In genome-wide RNAi screening of primary mouse neurons, SARM1 (STERILE ALPHA AND TIR motif-containing 1)) was identified, wherein the SARM1 was knocked down such that sensory neurons were protected over time against damage-induced axonal degeneration (Gerdts et al, j. Neurosci.,2013,33,13569-13580). SARM1 belongs to the cytoplasmic adaptor protein family but is unique among its members in that it is the oldest adaptor evolved, contradictively inhibits TLR signaling, and has been identified as a core executor of the injury-induced axonal 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, journal of neuroscience, 33,2013,13569-13580). Activation of SARM1 by axonal damage or forced dimerization of the SARM1-TIR domain promotes rapid and catastrophic consumption of nicotinamide adenine dinucleotide (NAD+) followed by rapid axonal degeneration, highlighting the core role of NAD+ homeostasis in axonal integrity. (Gerdts, J. Et al, science 2015,348,453-457). SARM1 is necessary for such damage-induced NAD+ depletion both in vivo and in vitro, and activation of SARM1 locally triggers axonal degeneration via destruction of NAD (+) (Gerdts et al, science 2015 348,452-457; sasaki et al, journal of biochemistry (J.biol. Chem.) 2015,290,17228-17238, both of which are hereby incorporated by reference in their entirety.

From studies of loss of genetic function, it is clear that SARM1 acts as a core executor of the axonal degeneration pathway after injury. Genetic knockout of SARM1 allows axons following nerve transection to be preserved for up to 14 days (Osterloh, J.M. et al, science 2012,337,481-484; gerdts, J. Et al, journal of neuroscience, 2013,33,13569-13580), and also improves functional outcome following traumatic Brain injury in mice (Henninger, N. et al, brain 139,2016,1094-1105). In addition to the role of SARM1 in direct axonal injury, SARM1 was also observed to be necessary for axonal degeneration in chemotherapy-induced peripheral neuropathy. The absence of SARM1 blocks chemotherapy-induced peripheral neuropathy, both of which inhibit axonal degeneration and produce increased pain sensitivity following chemotherapy vincristine treatment (Geisler et al, brain 2016,139,3092-3108).

SARM1 contains a number of conserved motifs, including SAM domains, ARM/HEAT motifs and TIR domains (FIG. 1), which mediate oligomerization and protein-protein interactions (O' Neill, L.A. and Bowie, A.G., nat.Rev.Immunol.,2007,7,353-364; tewari, R.et al, TRENDS CELL biol.,2010,20,470-481; qiao, F. And Bowie, J.U., sci.STKE,2005, re7, 2005). TIR domains are typically found in signaling proteins that play a role in the innate immune pathway, where they act as scaffolds for protein complexes (O' Neill, l.a. and Bowie, A.G., nat.Rev.Immunol.,2007,7,353-364). Interestingly, dimerization of the SARM1-TIR domain was sufficient to induce axonal degeneration and rapidly triggered degradation of NAD+ by acting as an NAD+ lyase (Milbrandt et al, WO 2018/057989; gerdts, J. Et al, science,2015,348,453-457). In view of the core role of SARM1 in the axonal degeneration pathway and its established NAD enzymatic activity, many efforts have been made to identify agents that can modulate SARM1 and potentially act as useful therapeutic agents, for example, to prevent the occurrence of neurodegenerative diseases, including peripheral neuropathy, traumatic brain injury, and/or neurodegenerative diseases.

The present disclosure provides certain compounds and/or compositions that are SARM1 inhibitors, and techniques related thereto.

Compounds of formula (I)

In a first aspect, the present disclosure provides a compound of formula I:

Or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof,

Wherein:

R ³ is- (CH ₂)_n Cy) and n is 0, 1 or 2;

Or alternatively

provided that the compound is not

In some embodiments, R ¹ is selected from the following groups:

a)-CN；

b)-NO₂；

c) -C (O) R ^1a, wherein R ^1a is as defined above, preferably optionally substituted C _1-6 aliphatic, more preferably optionally substituted C _1-6 alkyl, more preferably optionally substituted C _1-4 alkyl, even more preferably CH ₃;

d) -S (O) ₂R^1a, wherein R ^1a is as defined above, preferably optionally substituted C _1-6 aliphatic, more preferably optionally substituted C _1-6 alkyl, more preferably optionally substituted C _1-4 alkyl, even more preferably CH ₃;

e) -CONR ^1aR^1b, wherein R ^1a and R ^1b are each as defined above:

e1 In some such embodiments, R ¹ comprises-CONR ^1aR^1b, wherein R ^1a and R ^1b are each selected from hydrogen and optionally substituted C _1-6 aliphatic, preferably hydrogen and optionally substituted C _1-6 alkyl, more preferably H and optionally substituted C _1-4 alkyl, even more preferably hydrogen and CH ₃. In some embodiments, R ¹ includes-CONH ₂ and-CON (CH ₃)₂. In some embodiments, R ¹ also includes

E2 In other such embodiments, R ¹ comprises —conr ^1aR^1b, wherein R ^1a and R ^1b are each selected from hydrogen and an optionally substituted 3-to 7-membered saturated or partially unsaturated carbocyclic ring (preferably C _3-7 cycloalkyl). In some embodiments, R ¹ comprises

E3 In other such embodiments, R ¹ comprises-C (O) NR ^1aR^1b, wherein R ^1a and R ^1b are independently selected from optionally substituted C _1-6 aliphatic and optionally substituted phenyl, preferably optionally substituted C _1-6 alkyl and optionally substituted phenyl, more preferably optionally substituted C _1-4 alkyl and optionally substituted phenyl, even more preferably CH ₃ and phenyl. In some embodiments, R ¹ comprises

E4 In other such embodiments, R ¹ comprises-C (O) NR ^1aR^1b, wherein R ^1a and R ^1b together with the nitrogen atom to which they are attached form: i) 3 to 6 membered saturated or partially unsaturated monocyclic heterocycle, preferably 3 to 6 membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 nitrogen heteroatoms and 0 to 1 heteroatoms selected from oxygen and sulfur, in particular 6 membered saturated or partially unsaturated monocyclic heterocycle, preferably pyrrolidinyl ring, morpholinyl ring or piperidinyl ring; ii) an 8-to 10-membered saturated or partially unsaturated bridged bicyclic heterocycle, preferably an 8-to 10-membered saturated or partially unsaturated bridged bicyclic heterocycle having 1 or 2 nitrogen heteroatoms and 0-1 heteroatoms independently selected from oxygen and sulfur, such as a 2-azabicyclo [2.2.2] octyl ring. In some embodiments, R ¹ comprises

F) -S (O) ₂NR^1aR^1b, wherein R ^1a and R ^1b are each as defined above, preferably selected from hydrogen and optionally substituted C _1-6 aliphatic, more preferably hydrogen and optionally substituted C _1-6 alkyl, more preferably H and optionally substituted C _1-4 alkyl, even more preferably hydrogen and CH ₃. In some such embodiments, R ¹ comprises-S (O) ₂NH₂ and-S (O) ₂N(CH₃)₂;

g) -C (=nr ^1a)NR^1aR^1b, wherein R ^1a and R ^1b are each as defined above:

g1 In some such embodiments, R ¹ comprises-C (=nr ^1a)NR^1aR^1b) wherein R ^1a and R ^1b are each selected from hydrogen and optionally substituted C _1-6 aliphatic, preferably hydrogen and optionally substituted C _1-6 alkyl, more preferably H and optionally substituted C _1-4 alkyl, even more preferably hydrogen and CH ₃. In some embodiments, R ¹ comprises-C (=nh) NHCH ₃、-C(＝NH)N(CH₃)₂. In other embodiments, R ¹ comprises-C (=nr ^1a)NR^1aR^1b) wherein R ^1a and R ^1b are each selected from hydrogen and C _1-6 alkyl substituted with- (CH ₂)_0-4OR^○), preferably hydrogen and C _1-6 aliphatic substituted with-OH, in some embodiments, R ¹ comprises In other embodiments, R ¹ comprises-C (=nr ^1a)NR^1aR^1b) wherein R ^1a and R ^1b are each selected from H, CH ₃ and C _1-6 alkyl substituted with-OR ^○, in some embodiments, R ¹ comprises

G2 In other such embodiments, R ¹ comprises-C (=nh) NR ^1aR^1b, wherein R ^1a and R ^1b together with the nitrogen atom 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 comprises

H) -CO ₂R^1a, wherein R ^1a and R ^1b are each as defined above, preferably selected from hydrogen and optionally substituted C _1- ₆ aliphatic, more preferably hydrogen and optionally substituted C _1-6 alkyl, more preferably H and optionally substituted C _1-4 alkyl, even more preferably hydrogen and CH ₃. In some such embodiments, R ¹ comprises-CO ₂ H and-CO ₂CH₃;

i) 3 to 7 membered saturated or partially unsaturated carbocyclyl substituted with 0, 1,2, 3 or 4R ^x (preferably C _3-7 cycloalkyl);

j) C _6-10 aryl (e.g., phenyl or naphthyl) substituted with 0, 1,2, 3, or 4R ^x;

k) 5-or 6-membered heteroaryl substituted with 0, 1,2, 3 or 4R ^x having 1-3 (e.g., 1,2 or 3) heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably an unsubstituted 5-membered heteroaryl ring having 2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, more preferably And

L) 8-to 10-membered bicyclic heteroaryl substituted with 0, 1,2,3 or 4R ^x having 1-4 (e.g., 1,2 or 3) heteroatoms independently selected from nitrogen, oxygen and sulfur.

In some embodiments as described above, R ¹ is selected from -CN、-NO₂、-C(O)R^1a、-S(O)₂R^1a、-CONR^1aR^1b、-S(O)₂NR^1aR^1b、-C(＝NR^1a)NR^1aR^1b、-CO₂R^1a、C_3-7 cycloalkyl, 4-to 6-membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, phenyl, naphthyl, 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the cycloalkyl, heterocyclyl, phenyl, naphthyl and heteroaryl are each substituted with 0, 1,2, 3 or 4R ^x.

In some further embodiments, R ¹ is selected from -CN、-NO₂、-C(O)R^1a、-S(O)₂R^1a、-CONR^1aR^1b、-S(O)₂NR^1aR^1b、-C(＝NR^1a)NR^1aR^1b、-CO₂R^1a、 phenyl, a 5 or 6 membered heteroaryl having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the phenyl and heteroaryl are each substituted with 0, 1,2,3, or 4R ^x.

In some further embodiments, R ¹ is selected from: -CN,

In any of the above embodiments, R ² is selected from H, optionally substituted C _1-6 alkyl, -C (O) R ^2a、-S(O)₂R^2a, and-CO ₂R^2a. in some embodiments, R ² is selected from H, optionally substituted C _1-6 alkyl, -C (O) R ^2a, and-S (O) ₂R^2a. In any of the above embodiments, R ^2a is optionally substituted C _1-6 aliphatic, preferably optionally substituted C _1-6 alkyl, more preferably optionally substituted C _1-4 alkyl, even more preferably CH ₃. In some embodiments, R ² is-C (O) R ^2a, e.g., -C (O) CH ₃. In some embodiments, R ² is optionally substituted C _1-6 alkyl. In some preferred embodiments, R ² is H.

In any of the above embodiments, R ³ is-CH ₂ -Cy. In other embodiments, R ³ is- (CH ₂)₂ -Cy.) and in some preferred embodiments, R ³ is-Cy.

In any of the above embodiments, cy is selected from the group consisting of C _3-7 cycloalkyl, an 8-to 10-membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, C _6-10 aryl, a 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8-to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the cycloalkyl, heterocyclyl, aryl, and heteroaryl are each substituted with 0, 1,2,3, or 4R ^x.

In any of the above embodiments, R ⁴ is selected from the following groups:

a)H；

b) Optionally substituted C _1-6 aliphatic, preferably:

b1 Unsubstituted C _1-6 alkyl, more preferably C _1-3 alkyl, such as-CH ₃、-CH₂CH₃、-CH₂CH₂CH₃ or-CH (CH ₃)₂;

b2 C _1-6 aliphatic substituted with a group selected from: halogen 、-(CH₂)_0-4R^○、-(CH₂)_0-4OR^○、-(CH₂)_0-4N(R^○)₂、-(CH₂)_0-4S(O)₂R^○、-(CH₂)_0-4C(O)R^○、-(CH₂)_0-4C(O)N(R^○)₂ or- (CH ₂)_0- ₄C(O)OR^○).

In some such embodiments, R ⁴ includes a C _1-6 aliphatic substituted by- (CH ₂)_0-4R^○) in some such embodiments, R ^○ is a 5-to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, in some embodiments, R ⁴ includes a C _1-6 aliphatic substituted by phenyl, in some embodiments, R ⁴ includes a moiety substituted by phenylSubstituted C _1-6 aliphatic. In some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with a 5 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R ⁴ comprises quiltSubstituted C _1-6 aliphatic. In some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with a 6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R ⁴ comprises quiltSubstituted C _1-6 aliphatic. In some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with an 8-to 10-membered bicyclic heteroaryl group having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R ⁴ comprises quiltSubstituted C _1-6 aliphatic. In some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with a 5 membered saturated heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, R ⁴ comprises quiltSubstituted C _1-6 aliphatic.

In other such embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with- (CH ₂)_0-4OR^○), in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-OR ^○, in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-OH, in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-OCH ₃, in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-OCH ₂CH₂OCH₂ C≡CH.

In other such embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with- (CH ₂)_0-4N(R^○)₂). In some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-N (R ^○)₂. In some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-NH ₂. In some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-N (CH ₃)₂).

In other such embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with- (CH ₂)_0-4S(O)₂R^○). In some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-S (O) ₂R^○, in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-S (O) ₂CH₃.

In other such embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with- (CH ₂)_0-4C(O)N(R^○)₂).

In some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-C (O) N (R ^○)₂), in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-C (O) NH ₂, in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-C (O) N (R ^○)₂), wherein two independently occurring R ^○, taken together with their intermediate atoms, form a 3 to 12 membered saturated ring, partially unsaturated ring, or aryl monocyclic or bicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-C (O) N (R ^○)₂), wherein two independently occurring R ^○, taken together with their intermediate atoms, form a 5 to 6 membered saturated monocyclic ring having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, in some such embodiments, R ⁴ comprises a substituted aryl monocyclic or bicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfurSubstituted C _1-6 aliphatic. In some embodiments, R ⁴ comprises quiltSubstituted C _1-6 aliphatic. In some embodiments, R ⁴ comprises quiltSubstituted C _1-6 aliphatic.

In other such embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with- (CH ₂)_0-4C(O)OR^○), in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-C (O) OR ^○, in some embodiments, R ⁴ comprises a C _1-6 aliphatic substituted with-C (O) OH.

In any of the above embodiments, the C _1-6 aliphatic is preferably C _1-6 alkyl, more preferably C _1-3 alkyl, such as-CH ₃、-CH₂CH₃、-CH₂CH₂CH₃ or-CH (CH ₃)₂;

c) -C (O) R ^4a、-S(O)₂R^4a、-CO₂R^4a, wherein R ^4a is as defined above, preferably optionally substituted C _1-6 aliphatic, more preferably optionally substituted C _1-6 alkyl, more preferably optionally substituted C _1-4 alkyl, even more preferably CH ₃;

d) The following groups each substituted with 0, 1,2, 3 or 4R ^x:

d1 3 to 7 membered saturated or partially unsaturated carbocyclyl, preferably C _3-7 cycloalkyl;

d2 4 to 7 membered saturated or partially unsaturated monocyclic heterocyclyl having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably 4 to 6 membered saturated or partially unsaturated monocyclic heterocyclyl;

d3 C _6-10 aryl, preferably phenyl and naphthyl;

d4 A 5 or 6 membered heteroaryl group having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur.

In some further embodiments, R ⁴ is selected from hydrogen, optionally substituted C _1-6 aliphatic, -C (O) R ^4a、-S(O)₂R^4a、-CO₂R^4a、C_3-7 cycloalkyl, 4 to 6 membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5 or 6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl are each substituted with 0, 1,2,3 or 4R ^x.

In some embodiments, R ⁴ is H, -CH ₃、-CH₂CH₃、-CH₂CH₂CH₃、-CH(CH₃)₂, or a group selected from:

In any of the above embodiments, R ^x is selected from the following groups:

a) Halogen, -CN, -NO ₂, -OH and-SH;

b) -O-optionally substituted C _1-6 aliphatic and-S-optionally substituted C _1-6 aliphatic, wherein the optionally substituted C _1-6 aliphatic is preferably optionally substituted C _1-6 alkyl, more preferably optionally substituted C _1-4 alkyl, even more preferably CH ₃;

c)-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-NR^3aR^3b、-CONR^3aR^3b、-CO₂R^3a、-N(R^3a)SO₂R^3b and-N (R ^3a)C(O)R^3b, wherein R ^3a、R^3b is each as defined above, preferably selected from H and optionally substituted C _1-6 aliphatic, more preferably H and optionally substituted C _1-6 alkyl, more preferably H and optionally substituted C _1-4 alkyl, even more preferably H and CH ₃ in some such embodiments, R ^x comprises -COCH₃、-SO₂CH₃、-SO₂NHCH₃、-SO₂N(CH₃)₂、-NH₂、-NH(C_1-6 alkyl) (particularly -NHCH₃)、-CONH₂、-CO₂H、-CO₂CH₃、-NHSO₂CH₃、-N(CH₃)SO₂CH₃;

D) Optionally substituted C _1-6 aliphatic, preferably optionally substituted C _1-6 alkyl, more preferably optionally substituted C _1-4 alkyl, even more preferably optionally substituted CH ₃;

e) Optionally substituted 5-or 6-membered saturated or partially unsaturated heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur, e.g And

F) Optionally substituted 8-to 10-membered heteroaryl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably optionally substituted 9-membered heteroaryl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur, e.g.

In some further embodiments, each R ^x is independently selected from halogen, -CN, -NO ₂, -OH, -O-optionally substituted C _1-6 alkyl, -SH, -S-optionally substituted C _1-6 alkyl 、-NR^3aR^3b、-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a、-CONR^3aR^3b、-N(R^3a)SO₂R^3b、-N(R^3a)C(O)R^3b, and optionally substituted C _1-6 alkyl.

In some embodiments, R ³ is selected from

In any of the above embodiments, unless otherwise indicated, R ^1a、R^1b、R^2a、R^3a、R^3b and R ^4a may each independently be hydrogen or optionally substituted C _1-6 alkyl, optionally substituted phenyl, or optionally substituted C _3-7 cycloalkyl. In some embodiments, R ^1a、R^1b、R^2a、R^3a、R^3b and R ^4a may each independently be hydrogen or optionally substituted C _1-6 alkyl, more preferably H and optionally substituted C _1-4 alkyl, even more preferably H and CH ₃.

In any of the above embodiments, unless otherwise indicated, when applicable, R ^1a and R ^1b, and/or R ^3a and R ^3b, together with the nitrogen atom to which they are attached, form: i) 3 to 6 membered saturated or partially unsaturated monocyclic heterocycle, preferably 3 to 6 membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 nitrogen heteroatoms and 0 to 1 heteroatoms selected from oxygen and sulfur, in particular 6 membered saturated or partially unsaturated monocyclic heterocycle, preferably pyrrolidinyl ring, morpholinyl ring or piperidinyl ring; ii) an 8-to 10-membered saturated or partially unsaturated bridged bicyclic heterocycle, preferably an 8-to 10-membered saturated or partially unsaturated bridged bicyclic heterocycle having 1 or 2 nitrogen heteroatoms and 0-1 heteroatoms independently selected from oxygen and sulfur, such as a 2-azabicyclo [2.2.2] octyl ring.

In some embodiments, R ² and R ³ together with the nitrogen atom to which they are attached form a saturated or partially unsaturated 4 to 7 membered ring fused to Cy, or form a saturated or partially unsaturated 4 to 7 membered 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 group consisting of:

Wherein Cy is substituted with 0-4R ^x, R ^x and Cy are as defined in any of the embodiments described hereinabove.

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

In a second aspect, as a subset of the first aspect, the present disclosure provides a compound as described above, or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug, or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound, or prodrug thereof, wherein:

R ¹ is selected from -CN、-NO₂、-CONR^1aR^1b、-S(O)₂NR^1aR^1b、-CO₂R^1a、 phenyl and a 5 or 6 membered heteroaryl having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the phenyl and heteroaryl are each substituted with 0, 1,2,3 or 4R ^x;

R ² is selected from hydrogen, optionally substituted C _1-6 alkyl, -C (O) R ^2a, and-S (O) ₂R^2a;

R ³ is- (CH ₂)_n Cy) and n is 0, 1 or 2, preferably 0;

Cy is selected from C _3-7 cycloalkyl, 8 to 10 membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5 or 6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and 8 to 10 membered bicyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl are each substituted with 0, 1, 2,3 or 4R ^x;

r ⁴ is selected from hydrogen, optionally substituted C _1-6 alkyl, -C (O) R ^4a、-S(O)₂R^4a、C_3-7 cycloalkyl, 4-to 6-membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl are each substituted with 0, 1,2,3 or 4R ^x; and

Each R ^x is independently selected from halogen, -CN, -NO ₂, -OH, -O-optionally substituted C _1-6 alkyl, -SH, -S-optionally substituted C _1-6 alkyl 、-NR^3aR^3b、-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a、-CONR^3aR^3b, and optionally substituted C _1-6 alkyl;

R ^1a、R^1b、R^2a、R^3a、R^3b and R ^4a are each independently hydrogen, optionally substituted C _1-6 alkyl.

In any and all embodiments described above in accordance with the first and second aspects, unless otherwise indicated, R ¹ is selected from-CN, -NO ₂、-CONR^1aR^1b、-S(O)₂NR^1aR^1b, and a 5 or 6 membered heteroaryl having 1-3 (e.g., 1,2, or 3) heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the heteroaryl is substituted with 0, 1,2,3, or 4R ^x.

In some embodiments, R ¹ is selected from-CONR ^1aR^1b and-S (O) ₂NR^1aR^1b, particularly-CONR ^1aR^1b. In some such embodiments, R ^1a and R ^1b are each independently H or optionally substituted C _1-4 alkyl, preferably H or unsubstituted C _1-4 alkyl, even more preferably H or CH ₃. In some embodiments R ¹ is selected from the group consisting of-CONH ₂ and-CON (CH ₃)₂).

In some preferred embodiments, R ¹ is selected from —cn and an unsubstituted 5 membered heteroaryl ring having 2 or 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some more preferred embodiments, R ¹ is-CN,More preferably-CN orEven more preferably-CN.

In any and all embodiments described above in accordance with the first and second aspects, unless otherwise indicated, R ² is selected from hydrogen, optionally substituted C _1-6 alkyl, -C (O) R ^2a, and-S (O) ₂R^2a. In some embodiments, R ² is-C (O) R ^2a. In some of the above embodiments, R ^2a is independently optionally substituted C _1-4 alkyl, preferably unsubstituted C _1-4 alkyl, more preferably CH ₃. In some embodiments, R ² is-C (O) CH ₃.

In some preferred embodiments, R ² is selected from hydrogen and optionally substituted C _1-4 alkyl (preferably unsubstituted C _1- ₄ alkyl, especially CH ₃), more preferably H.

In some preferred embodiments, R ¹ is —cn, and R ² is H.

In any and all embodiments described above in accordance with the first and second aspects, unless otherwise indicated, R ³ is- (CH ₂)_n Cy) and n is 0, thus, in some preferred embodiments, R ³ is Cy.

In any and all embodiments described above in accordance with the first and second aspects, unless otherwise indicated, cy is selected from C _3-6 cycloalkyl, an 8-to 10-membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, phenyl, naphthyl, a 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and an 8-to 10-membered bicyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the cycloalkyl, heterocyclyl, phenyl, naphthyl and heteroaryl are each substituted with 0, 1, 2,3 or 4R ^x.

In some such embodiments, cy includes C _3-6 cycloalkyl substituted with 0,1,2, or 3R ^x, particularly unsubstituted C _3-6 cycloalkyl. In some further embodiments, cy comprises unsubstituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

In some preferred embodiments, cy comprises phenyl substituted with 0, 1,2, 3 or 4, preferably 0, 1,2 or 3R ^x. In some embodiments, cy comprises unsubstituted phenyl. In some embodiments, cy comprises phenyl substituted with 1R ^x, particularly at the 2-position. In some embodiments, cy includes phenyl substituted with 2R ^x, particularly at the 2-and 3-positions, the 2-and 5-positions, or the 2-and 6-positions. In some embodiments, cy includes phenyl substituted with 3R ^x, particularly at the 2-, 3-, and 4-positions, or the 2-, 3-, and 5-positions, or the 2-, 3-, and 6-positions. In some preferred embodiments, cy comprises:

preferred embodiments are those wherein the phenyl group is unsubstituted or substituted with 1 or 2R ^x.

In other preferred embodiments, cy comprises a naphthalene group substituted with 0, 1, 2,3 or 4, preferably 0, 1, 2 or 3R ^x. In some embodiments, cy comprises unsubstituted naphthyl. In some embodiments, cy comprises naphthyl substituted with 1R ^x. In some embodiments, cy comprises naphthyl substituted with 2R ^x. In some embodiments, cy comprises naphthyl substituted with 3R ^x. In some embodiments, cy comprises:

In some preferred embodiments, cy is More preferably

In other preferred embodiments, cy comprises a 5 or 6 membered heteroaryl group having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with 0, 1,2, 3, or 4, preferably 0, 1,2, or 3R ^x. In some embodiments, cy comprises a 5 membered heteroaryl group having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, cy comprises a 6 membered heteroaryl having 1-3 nitrogen heteroatoms. In some embodiments, cy comprises a 6 membered heteroaryl ring having 1-2 nitrogen heteroatoms. In some embodiments, cy comprises pyridinyl. In some such embodiments, cy comprises pyridin-2-yl, preferably pyridin-3-yl or pyridin-4-yl. In some embodiments, cy includes a pyridazinyl group, particularly a pyridazin-4-yl group. In some embodiments, cy comprises pyrazinyl. In some embodiments, cy comprises pyrimidinyl. In any of the above embodiments, cy is substituted with 1 or 2R ^x. In some embodiments, cy comprises:

preferably

In other preferred embodiments Cy comprises an 8-to 10-membered bicyclic heteroaryl group having 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, particularly an 8-membered, particularly 9-or 10-membered bicyclic heteroaryl group having 1,2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, substituted with 0, 1,2,3 or 4, preferably 0, 1,2 or 3R ^x. In some such embodiments, the bicyclic heteroaryl is a fused ring system of a benzene ring and a 5 or 6 membered heteroaryl, wherein the heteroatoms are all in the 5 or 6 membered heteroaryl ring and are not common atoms. In some preferred embodiments, cy comprises a 9 or 10 membered bicyclic heteroaryl group having 1-2 nitrogen heteroatoms. In some embodiments, cy comprises a 9 membered bicyclic heteroaryl having 1 or 2 nitrogen heteroatoms. In some embodiments, cy comprises a 10 membered bicyclic heteroaryl having 1 or 2 nitrogen heteroatoms. In some further embodiments, the bicyclic heteroaryl has only nitrogen atoms as heteroatoms. In some embodiments, cy comprises indolyl, indazolyl, benzimidazolyl, quinolinyl, or quinazolinyl. In some embodiments, cy includes indol-3-yl, indol-4-yl, indol-5-yl, indol-6-yl or indol-7-yl. In some embodiments, cy includes indazol-3-yl, indazol-4-yl, indazol-5-yl, indazol-6-yl, or indazol-7-yl. In some embodiments, cy includes quinolin-2-yl, quinolin-3-yl, quinolin-4-yl, quinolin-5-yl, quinolin-6-yl, quinolin-7-yl, or quinolin-8-yl. In some preferred embodiments, cy is substituted with 1R ^x.

In some particular embodiments, cy comprises:

preferably

In other preferred embodiments, cy comprises an 8-to 10-membered (especially 9-or 10-membered) bicyclic heterocyclyl having 1-3 (e.g., 1,2, or 3) heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with 0,1,2, or 3R ^x. In some such embodiments, the bicyclic heterocyclyl is a fused ring system of a benzene ring with a 5 or 6 membered saturated or partially unsaturated heterocyclyl ring, wherein the heteroatoms are all in the 5 or 6 membered saturated or partially unsaturated heterocyclyl ring and are not common atoms. In some embodiments, the bicyclic heterocyclyl is a 9 or 10 membered saturated or partially unsaturated bicyclic heterocyclyl having 1 or 2 heteroatoms selected independently from nitrogen, oxygen and sulfur. In some embodiments, the bicyclic heterocyclyl is a 9 or 10 membered saturated or partially unsaturated bicyclic heterocyclyl having 1 or 2 heteroatoms selected independently from oxygen and sulfur. In some preferred embodiments, the bicyclic heterocyclyl is a 9 or 10 membered saturated or partially unsaturated bicyclic heterocyclyl having 1 or 2, preferably 1, oxygen heteroatoms. In some embodiments, the bicyclic heterocyclyl is a 9 membered saturated or partially unsaturated bicyclic heterocyclyl having 1 or 2, preferably 1, sulfur heteroatoms. In some embodiments, the bicyclic heterocyclyl is a 9 or 10 membered saturated or partially unsaturated bicyclic heterocyclyl having 1 or 2 nitrogen heteroatoms and 0 or 1 oxygen or sulfur heteroatoms. In some preferred embodiments, the bicyclic heterocyclyl is a 9 membered saturated or partially unsaturated bicyclic heterocyclyl having 1 nitrogen heteroatom and 0 or 1, preferably 0 oxygen or sulfur heteroatoms. In some embodiments, cy includes 2, 3-dihydrobenzofuranyl, 2, 3-dihydrobenzothienyl, indolinyl, 2, 3-dihydrobenzindazolyl, 2, 3-dihydrobenzimidazolyl, chromanyl, thiochroman, tetrahydroquinolinyl. In some embodiments, cy includes 2, 3-dihydrobenzofuranyl-4-yl, 2, 3-dihydrobenzofuranyl-6-yl. In some embodiments, cy includes 2, 3-dihydrobenzothiophen-4-yl, 2, 3-dihydrobenzothiophen-6-yl. In some embodiments, cy includes indolin-4-yl, indolin-6-yl. In some such embodiments, cy is substituted with 1R ^x. In some embodiments, cy comprises:

Preferably is

In some preferred embodiments, cy is selected from:

Phenyl unsubstituted or substituted with 1,2 or 3R ^x as described above;

Naphthyl substituted with 1R ^x;

A 5-or 6-membered heteroaryl group having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably a 5-membered heteroaryl group having 1 nitrogen heteroatom and 0 or 1 heteroatom selected from nitrogen, oxygen and sulfur, more preferably a 6-membered heteroaryl group having 1 or 2 nitrogen heteroatoms (especially pyridyl, pyridazinyl or pyrimidinyl), more preferably pyridin-2-yl, even more preferably pyridin-3-yl, pyridin-4-yl or pyridazin-4-yl, each of which is substituted with 1 or 2R ^x;

A 9 or 10 membered bicyclic heteroaryl group having 1,2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, more preferably a fused bicyclic heteroaryl group of a benzene ring and a 5 or 6 membered heteroaryl group as described above, even more preferably a 9 or 10 membered bicyclic heteroaryl group having 1-2 nitrogen heteroatoms (particularly quinolinyl and indazolyl), each of which groups is substituted with 0, 1,2 or 3R ^x; and

A 9 or 10 membered saturated or partially unsaturated bicyclic heterocyclic group having 1,2 or 3, preferably 1 or 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably a fused bicyclic heterocyclic group of a benzene ring and a 5 or 6 membered saturated or partially unsaturated heterocyclic group as described above, more preferably a 9 or 10 (particularly 9) membered saturated or partially unsaturated bicyclic heterocyclic group having 1 or 2 (particularly 1) heteroatoms independently selected from oxygen and sulfur, or a 9 or 10 (particularly 9) membered saturated or partially unsaturated bicyclic heterocyclic group having 1 or 2 (particularly 1) nitrogen heteroatoms and 0 or 1 (particularly 0) oxygen or sulfur heteroatoms, each of these groups being substituted with 0,1,2 or 3R ^x;

More preferably selected from:

preferably Preferably

In any and all embodiments described above in accordance with the first and second aspects, unless otherwise indicated, R ⁴ is selected from the following groups:

a)H；

b2 C _1-6 alkyl (preferably C _1-4 alkyl) substituted with a group selected from: -OH, -SH, and 1,2, 3 or more halogens.

In some such embodiments, R ⁴ comprises C _1-6 alkyl substituted with 1,2,3 or more halogens, e.g., (methyl substituted with 1,2 or preferably 3 halogens) -C _0-5 alkyl-, e.g., (methyl substituted with 1,2 or preferably 3 halogens) -C _0-3 alkyl-. In some embodiments, the halogen is F, cl or Br, preferably F or Cl, more preferably F. In some embodiments, R ⁴ comprises-CF ₃、-CH₂CF₃、-CH₂CH₂CF₃ or-CH ₂CH₂CH₂CF₃.

In other such embodiments, R ⁴ comprises an OH-substituted C _1-6 alkyl (preferably C _1-4 alkyl), such as-CH ₂-OH、-CH₂CH₂-OH、-CH₂CH₂CH₂ -OH or-CH ₂CH₂CH₂CH₂ -OH. In other such embodiments, R ⁴ comprises SH substituted C _1-6 alkyl (preferably C _1-4 alkyl), such as-CH ₂-SH、-CH₂CH₂-SH、-CH₂CH₂CH₂ -SH or-CH ₂CH₂CH₂CH₂ -SH;

c) -C (O) R ^4a and-S (O) ₂R^4a, wherein R ^4a is as defined above, preferably optionally substituted C _1-6 alkyl, more preferably optionally substituted C _1-4 alkyl, even more preferably CH ₃;

d) The following groups each substituted with 0, 1,2, 3 or 4R ^x:

d1 C _3-7 cycloalkyl, preferably C _4-6 cycloalkyl, such as cyclobutyl, cyclopentyl and cyclohexyl;

d2 A 4,5 or 6 membered saturated or partially unsaturated monocyclic heterocyclyl group having 1,2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, such as azetidinyl;

d3 C _6-10 aryl, preferably phenyl and naphthyl;

d4 A 5 or 6 membered heteroaryl group having 1,2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, such as pyrazolyl;

preferably, the cycloalkyl, heterocyclyl, aryl and heteroaryl groups are unsubstituted.

In some embodiments, R ⁴ is H、-CH₃、-CH₂CH₃、-CH₂CH₂CH₃、-CH(CH₃)₂、-CF₃、-CH₂CF₃、-CH₂CH₂CF₃、-CH₂CH₂CH₂CF₃、-CH₂-OH、-CH₂CH₂-OH、-CH₂CH₂CH₂-OH、-C(O)CH₃、-S(O)₂CH₃、

In some embodiments, when R ⁴ is as defined above, R ¹ is-CN. In some embodiments, R ⁴ is H and R ¹ is-CONR ^1aR^1b or a 5 or 6 membered heteroaryl as defined above with respect to R ¹, e.g., -CONH ₂、-CON(CH₃)₂, or

In any and all embodiments described above in accordance with the first and second aspects, unless otherwise indicated, R ^x is selected from the following groups:

a) Halogen (e.g., F, cl or Br), -CN, -NO ₂, -OH, and-SH;

b) -O-optionally substituted C _1-4 alkyl and-S-optionally substituted C _1-4 alkyl, preferably-O-CH ₃ and-S-CH ₃;

c)-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-NR^3aR^3b、-CONR^3aR^3b and-CO ₂R^3a, wherein R ^3a、R^3b is each as defined above, preferably selected from H and optionally substituted C _1-6 alkyl, more preferably H and optionally substituted C _1-4 alkyl, even more preferably H and CH ₃. In some such embodiments, R ^x comprises -COCH₃、-SO₂CH₃、-SO₂NHCH₃、-SO₂N(CH₃)₂、-NH₂、-NH(C_1-6 alkyl) (particularly —nhch ₃)、-CONH₂、-CO₂ H or-CO ₂CH₃.

D) C _1-6 alkyl (preferably C _1-4 alkyl, such as CH ₃) which is unsubstituted or substituted by 1, 2,3 or more halogens, for example CF ₃.

In some preferred embodiments, R ^x is selected from F, cl, -CN, CF ₃、-O-CH₃、-SO₂CH₃, or-CO ₂CH₃.

In any and all embodiments described above in accordance with the first and second aspects, unless otherwise indicated, R ³ is selected from:

in a third aspect, as a subset of either the first and second aspects, the present disclosure provides a compound as described above, or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof,

Wherein:

r ¹ is-CN;

r ² is hydrogen;

R ³ is Cy;

Cy is as defined in any of the embodiments above according to the first aspect, particularly the second aspect, preferably selected from 8 to 10 membered saturated or partially unsaturated bicyclic heterocyclyl having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5 or 6 membered heteroaryl having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur and 8 to 10 membered bicyclic heteroaryl having 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the heterocyclyl, aryl and heteroaryl are as defined in any of the embodiments above according to the first aspect, particularly the second aspect, and wherein the heterocyclyl, aryl and heteroaryl are each substituted with 0, 1, 2 or 3R ^x;

R ⁴ is selected from hydrogen, optionally substituted C _1-6 alkyl, and C _4-6 cycloalkyl substituted with 0,1,2, or 3R ^x; and

Each R ^x is independently selected from halogen, -CN, -NO ₂, -OH, -O-optionally substituted C _1-6 alkyl, -SH, -S-optionally substituted C _1-6 alkyl 、-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a、-CONR^3aR^3b, and optionally substituted C _1-6 alkyl;

R ^3a and R ^3b are each independently hydrogen or optionally substituted C _1-6 alkyl.

In some preferred embodiments, cy is phenyl substituted with 0,1, 2, or 3R ^x. In some embodiments, cy is unsubstituted phenyl. In some embodiments, cy is phenyl substituted with 1R ^x, particularly at the 2-position. In some embodiments, cy is phenyl substituted with 2R ^x, particularly at the 2-and 3-positions, the 2-and 5-positions, or the 2-and 6-positions. In some embodiments, cy is phenyl substituted with 3R ^x, particularly at the 2-, 3-, and 4-positions, or the 2-, 3-, and 5-positions, or the 2-, 3-, and 6-positions. In some preferred embodiments, cy is:

In some preferred embodiments, cy is naphthyl substituted with 1R ^x. In some preferred embodiments, cy is Preferably is

In some preferred embodiments Cy is a 5 or 6 membered heteroaryl group having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably a 5 membered heteroaryl group having 1 nitrogen heteroatom and 0 or 1 heteroatom selected from nitrogen, oxygen and sulfur, more preferably a 6 membered heteroaryl group having 1 or 2 nitrogen heteroatoms (especially pyridyl, pyridazinyl or pyrimidinyl), more preferably pyridin-2-yl, even more preferably pyridin-3-yl, pyridin-4-yl or pyridazin-4-yl, each of which is substituted with 1 or 2R ^x. In some preferred embodiments, cy is:

preferably

In some preferred embodiments, cy is a 9 or 10 membered bicyclic heteroaryl group having 1,2, or 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, substituted with 1,2, or 3R ^x, wherein the bicyclic heteroaryl group is a fused ring system of a benzene ring and a 5 or 6 membered heteroaryl group, wherein the heteroatoms are all in the 5 or 6 membered heteroaryl ring and are not common atoms. In some such embodiments, cy is preferably a 9 or 10 membered bicyclic heteroaryl group having 1-2 nitrogen heteroatoms, such as indolyl, indazolyl, benzimidazolyl, quinolinyl, or quinazolinyl. In some preferred embodiments, cy is a 9 membered bicyclic heteroaryl having 1 or 2 nitrogen atoms. In some embodiments, cy is a 10 membered bicyclic heteroaryl having 1 or 2 nitrogen atoms. In some preferred embodiments, cy is substituted with 1R ^x. In some preferred embodiments, cy isPreferably

In some preferred embodiments, cy is a 9 or 10 membered saturated or partially unsaturated bicyclic heterocyclic group having 1,2 or 3, preferably 1 or 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, substituted with 0, 1,2 or 3R ^x, wherein the bicyclic heterocyclic group is a fused ring system of a benzene ring and a 5 or 6 membered saturated or partially unsaturated heterocyclic group, wherein the heteroatoms are all in the 5 or 6 membered saturated or partially unsaturated heterocyclic ring and are not common atoms. Such bicyclic heterocyclic groups include 2, 3-dihydrobenzofuranyl, 2, 3-dihydrobenzothienyl, indolinyl, 2, 3-dihydrobenzindazolyl, 2, 3-dihydrobenzimidazolyl, chromanyl, thiochroman and tetrahydroquinolinyl. In some such embodiments, the bicyclic heterocyclyl is preferably a 9 or 10 (particularly 9) membered saturated or partially unsaturated bicyclic heterocyclyl having 1 or 2 (particularly 1) heteroatoms independently selected from oxygen and sulfur, preferably 2, 3-dihydrobenzofuranyl, particularly 2, 3-dihydrobenzofuran-4-yl; or a 9-or 10-membered saturated or partially unsaturated bicyclic heterocyclic group having 1 or 2 (in particular 1) nitrogen heteroatoms and 0 or 1 (in particular 0) oxygen or sulfur heteroatoms, preferably indolin-4-yl. In some preferred embodiments, cy is substituted with 1R ^x. In some preferred embodiments, cy is:

preferably

In some preferred embodiments, R ⁴ is selected from the following groups:

a)H；

b2 C _1-4 alkyl substituted with a group selected from: -OH and 1,2,3 or more halogens.

In some preferred embodiments, R ⁴ is C _1-4 alkyl substituted with 1,2,3 or more halogens, e.g., (methyl substituted with 1,2 or preferably 3 halogens) -C _0-5 alkyl-, e.g., (methyl substituted with 1,2 or preferably 3 halogens) -C _0-3 alkyl-. In some embodiments, R ⁴ is-CF ₃、-CH₂CF₃、-CH₂CH₂CF₃ or-CH ₂CH₂CH₂CF₃. In some preferred embodiments, R ⁴ is OH-substituted C _1-6 alkyl (preferably C _1-4 alkyl), such as-CH ₂-OH、-CH₂CH₂-OH、-CH₂CH₂CH₂ -OH or-CH ₂CH₂CH₂CH₂ -OH;

d1 Unsubstituted C _4-6 cycloalkyl groups such as cyclobutyl, cyclopentyl and cyclohexyl.

In some preferred embodiments, R ⁴ is H、-CH₃、-CH₂CH₃、-CH₂CH₂CH₃、-CH(CH₃)₂、-CF₃、-CH₂CF₃、-CH₂CH₂CF₃、-CH₂CH₂CH₂CF₃、-CH₂-OH、-CH₂CH₂-OH、-CH₂CH₂CH₂-OH、

In some preferred embodiments, R ^x is selected from the following groups:

a) Halogen (e.g., F, cl or Br), -CN;

c) -C (O) R ^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a and-CONR ^3aR^3b, wherein R ^3a、R^3b are each independently H or optionally substituted C _1-6 alkyl, preferably H or optionally substituted C _1-4 alkyl, more preferably H or CH ₃; and

D) C _1-6 alkyl substituted with 1,2, 3 or more halogens, preferably CF ₃.

In some more preferred embodiments, R ^x is selected from the following groups:

a)F、Cl、-CN；

b) -O-optionally substituted C _1-4 alkyl, preferably-O-CH ₃; and

C) -SO ₂R^3a and-CO ₂R^3a, wherein R ^3a is independently H or optionally substituted C _1-6 alkyl, preferably H or CH ₃. In some preferred embodiments, R ^x is-SO ₂CH₃ or-CO ₂CH₃.

D) C _1-4 alkyl substituted with 1,2, 3 or more halogens, preferably CF ₃.

In some embodiments, R ³ is selected from:

it is to be understood that the compounds of formula I have the following structure

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

Thus, it is understood that when R ⁴ is H, the compounds of formula I can be drawn in either tautomeric form.

In a fourth aspect, the present disclosure provides a compound of formula I according to the first, second and third aspects above, having a structure as shown in one of the following formulas I-a, I-a-I, I-a-ii or I-a-iii to I-a-xiv,

Or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof.

In some embodiments, R ¹ is —cn. Thus, in some embodiments, the present disclosure provides compounds of formula I-a:

Or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof, wherein each of R ²、R³ and R ⁴ is as defined in any embodiment hereinabove and described herein.

In some embodiments of formula I-a, R ² is H. Accordingly, the present disclosure provides compounds of formula I-a-I:

Or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof, wherein each of R ³ and R ⁴ is as defined in any embodiment hereinabove and described herein.

In some embodiments of formulas I-a-I, R ³ is Cy. Accordingly, the present disclosure provides compounds of formula I-a-ii:

or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof, wherein each of Cy and R ⁴ is as defined in any embodiment hereinabove and described herein.

In some embodiments of formulas I-a-ii, the present disclosure provides compounds of formulas I-a-iii through I-a-xiv:

or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof, wherein each of R ⁴ and R ^x is as defined in any embodiment hereinabove and described herein.

In a fifth aspect, as a subset of any of the first, second, third and fourth aspects, the present disclosure provides a compound of formula I as described above, or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof, wherein:

r ¹ is-CN;

r ² is hydrogen;

R ³ is Cy;

Cy is phenyl substituted with 1 or 2R ^x;

R ⁴ is selected from hydrogen and optionally substituted C _1-6 alkyl; and

Each R ^x is independently selected from halogen, -CN and optionally substituted C _1-6 alkyl.

Such compounds may be represented by the structure of formula I-a-xv, which is a subset of compounds of formula I-a-iii:

Wherein the method comprises the steps of

R ⁴ is selected from hydrogen and optionally substituted C _1-6 alkyl; and

In some preferred embodiments, cy is phenyl substituted with 1R ^x, particularly at the 2-position. In some preferred embodiments, cy is phenyl substituted with 2R ^x, particularly at the 2-and 3-positions, the 2-and 5-positions, or the 2-and 6-positions. In some preferred embodiments, cy is:

In some preferred embodiments, R ⁴ is selected from the following groups:

a)H；

b2 C _1-4 alkyl substituted with 1,2,3 or more halogens, for example-CF ₃、-CH₂CF₃、-CH₂CH₂CF₃ or-CH ₂CH₂CH₂CF₃, preferably-CF ₃、-CH₂CF₃、-CH₂CH₂CF₃.

In some preferred embodiments, R ⁴ is H, -CH ₃、-CF₃、-CH₂CF₃、-CH₂CH₂CF₃.

In some preferred embodiments, R ^x is selected from: halogen (e.g., F, cl or Br); -CN; c _1-6 alkyl substituted with 1,2, 3 or more halogens, preferably CF ₃.

In some preferred embodiments, R ^x is selected from F, cl, -CN, and CF ₃.

In some embodiments, R ³ is selected from:

in any of the embodiments described above, halogen is F, cl or Br, preferably F or Cl.

In some embodiments, the present disclosure provides a compound selected from the group consisting of:

Tool compounds

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

In some embodiments, the present disclosure provides compounds of formula II:

Or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof, wherein each of R ¹、R² and R ³ is as defined above with respect to formula I and subsets thereof, and is described in various aspects and subsets thereof; t is a bivalent tether moiety; and R ^t is a detectable moiety.

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

As used herein, the term "detectable moiety" is used interchangeably with the terms "label" and "reporter molecule" and refers to any moiety capable of being detected, such as a primary label and a secondary label. The presence of a detectable moiety can be measured by using a method that quantifies (in absolute, approximate, or relative fashion) 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 reporter moieties (e.g., labels, dyes, photocrosslinkers, cytotoxic compounds, drugs, affinity labels, photoaffinity labels, reactive compounds, antibodies or antibody fragments, biological materials, nanoparticles, spin labels, chromophores, metal-containing moieties, radioactive moieties, quantum dots, novel functional groups, groups that interact covalently or non-covalently with other molecules, photocleavable moieties, actinic radiation excitable moieties, ligands, photoisomerizable moieties, biotin analogs (e.g., biotin sulfoxides), moieties incorporating heavy atoms, chemically cleavable groups, photocleavable groups, redox active agents, isotopic labeled moieties, biophysical probes, phosphorescent groups, chemiluminescent groups, electronically dense groups, magnetic groups, intercalating groups, chromophores, energy transfer agents, bioactive agents, detectable labels, and any combination of the foregoing).

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

The term "secondary label" as used herein refers to moieties such as biotin and various protein antigens that require the presence of a second intermediate to produce a detectable signal. For biotin, the second intermediate may comprise a streptavidin-enzyme conjugate. For antigen labeling, the second intermediate may comprise an antibody-enzyme conjugate. Some fluorescent groups act as secondary labels because they transfer energy to another group during non-radiative Fluorescence Resonance Energy Transfer (FRET), while the second group generates a detection signal.

The terms "fluorescent label", "fluorescent dye" and "chromophore" as used herein 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 dye (Alexa Fluor 350、Alexa Fluor 488、Alexa Fluor 532、Alexa Fluor 546、Alexa Fluor568、Alexa Fluor 594、Alexa Fluor 633、Alexa Fluor 660 and Alexa Fluor 680), AMCA-S, BODIPY dye (BODIPY FL、BODIPY R6G、BODIPY TMR、BODIPY TR、BODIPY493/503、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 dye (Cy 3, cy 5), Cy3.5, cy5.5), dansyl, dapoxyl, dialkylaminocoumarin, 4',5' -dichloro-2 ',7' -dimethoxy-fluorescein, DM-NERF, eosin, erythrosin, fluorescein, FAM, hydroxycoumarin, IRDye (IRD 40, IRD 700, IRD 800), JOE, lissamine rhodamine (LISSAMINE RHODAMINE) B, sea blue (MarinaBlue), methoxycoumarin, naphthalene fluorescein, oregon Green 488, oregon Green 500, Oregon green 514, pacific Blue (Pacific Blue), pyMPO, pyrene, rhodamine B, rhodamine 6G, rhodamine green, rhodamine Red, luo Deer green (Rhodol Green), 2',4',5',7' -tetra-bromosulfone-fluorescein, tetramethyl-rhodamine (TMR), carboxytetramethyl rhodamine (TAMRA), texas Red (Texas Red), texas Red-X, 5 (6) -carboxyfluorescein, 2, 7-dichlorofluorescein, N-bis (2, 4, 6-trimethylphenyl) -3,4:9, 10-perylene bis (dicarboximide), HPTS, ethyl eosin, DY-490XLMegaStokes, DY-485XL MegaStokes, addisondack Green 520, ATTO465, ATTO 488, ATTO 495, YOYO-1, 5-FAM, BCECF, dichlorofluorescein, rhodamine 110, rhodamine 123, YO-PRO-1, green cyanine SYTOX (SYTOX Green), tissue Sodium fluorescence (Sodium Green), SYBR Green I, AlexaFluor 500, FITC, fluo-3, fluo-4, fluoro-emerald (fluoro-emerald), yoYo-1ssDNA, yoYo-1dsDNA, yoYo-1, SYTO RNASelect, di Wen Salu (DIVERSA GREEN) -FP, deragong Green (Dragon Green), aewa Green (EvaGreen), siteff Green (Surf Green) EX, spectral Green (Spectrum Green)、NeuroTrace 500525、NBD-X、MitoTracker Green FM、LysoTracker Green DND-26、CBQCA、PA-GFP( after activation), WEGFP (after activation), flASH-CCXXCC, thistle green monomer (Azami Green monomeric), thistle green, green Fluorescent Protein (GFP), EGFP (Campbell Tsien 2003), EGFP (Patterson 2001), kdeGreen (KAEDE GREEN), 7-benzylamino-4-nitrobenzo-2-oxa-1, 3-diazole, bexl, rubus parvifolius (Doxokumicin), lu Miao green (LumioGreen) and SuperGlo GFP.

The term "mass label" as used herein refers to any moiety capable of being uniquely detected by its mass using Mass Spectrometry (MS) detection techniques. Examples of mass labels include electrophoretic release labels such as N- [3- [4'- [ (p-methoxytetrafluorobenzyl) oxy ] phenyl ] -3-methyl glyceryl ] isopiperidinecarboxylic acid, 4' - [2,3,5, 6-tetrafluoro-4- (pentafluorophenoxy) ] methylacetophenone, and derivatives thereof. The synthesis and utility of these mass tags is described in U.S. Pat. nos. 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 labels include, but are not limited to, nucleotides, dideoxynucleotides, oligonucleotides of varying length and base composition, oligopeptides, oligosaccharides and other synthetic polymers of varying length and monomer composition. Various neutral and charged organic molecules (biomolecules or synthetic compounds) of suitable mass range (100-2000 daltons) can also be used as mass labels. Stable isotopes (e.g., ¹³C、²H、¹⁷O、¹⁸ O and ¹⁵ N) may also be used as mass tags.

The term "chemiluminescent group" as used herein refers to a group that emits light due to a chemical reaction without the addition of heat. For example, luminol (luminol, 5-amino-2, 3-dihydro-1, 4-phthalhydrazide) is reacted with an oxidizing agent such as hydrogen peroxide (H ₂O₂) in the presence of alkali and metal catalysts to produce the excited state product (3-aminophthalic acid, 3-APA).

The term "chromophore" as used herein refers to a molecule that absorbs light at the visible, UV, or IR wavelengths.

The term "dye" as used herein refers to a soluble coloring matter substance containing a chromophore.

The term "electron dense group" as used herein 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, anhydrous indium trichloride, 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 protein (Ag assay: 8.0-8.5%) "Strong" (Strength), silver tetraphenylporphine (S-TPPS), sodium chloroaurate, sodium tungstate, thallium nitrate, thiosemicarbazide (TSC), uranyl acetate, uranyl nitrate, and vanadyl sulfate.

The term "energy transfer agent" as used herein refers to a molecule that is directed to or receives energy from another molecule. By way of example only, fluorescence Resonance Energy Transfer (FRET) is a dipole-dipole coupling process by which excited state energy of a fluorescent donor molecule is transferred nonradiatively to an unexcited acceptor molecule, which then emits supplied energy in fluorescence at a longer wavelength.

The term "moiety incorporating a heavy atom" as used herein refers to a group incorporating ions of atoms generally heavier than carbon. In some embodiments, such ions or atoms include, but are not limited to, silicon, tungsten, gold, lead, and uranium.

The term "photoaffinity label" as used herein refers to a label bearing a group that, upon exposure to light, forms a link with a molecule having affinity for the label.

The term "photocage moiety" as used herein refers to a group that, upon irradiation at a certain wavelength, binds, covalently or non-covalently, other ions or molecules.

The term "photoisomerizable moiety" as used herein refers to a group that changes from one isomeric form to another when irradiated with light.

The term "radioactive moiety" as used herein refers to a group whose nuclei spontaneously emit nuclear radiation (e.g., alpha, beta, or gamma particles); wherein the alpha particles are helium nuclei, the beta particles are electrons, and the gamma particles are high energy photons.

The term "spin label" as used herein refers to a molecule containing an atom or group of atoms that exhibits unpaired electron spin (i.e., a stable paramagnetic group), which in some embodiments is detected by electron spin resonance spectroscopy, and in other embodiments is attached to another molecule. Such spin labeling molecules include, but are not limited to, nitroxyl radicals and nitroxides, and in some embodiments are either single spin labeling or dual spin labeling.

The term "quantum dot" as used herein refers to colloidal semiconductor nanocrystals, which in some embodiments are detected in the near infrared and have extremely high quantum yields (i.e., are very bright under moderate irradiation).

One of ordinary skill in the art will recognize that the detectable moiety may be attached to the provided compounds by suitable substituents. As used herein, the term "suitable substituent" refers to a moiety that is capable of covalently linking to a 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, sulfhydryl moieties or hydroxyl moieties, to name a few. It will be appreciated that such moieties may be attached to the provided compounds directly or through a tether moiety (e.g., a divalent saturated or unsaturated hydrocarbon chain).

In some embodiments, -T-is selected from the group consisting of- (CH ₂CH₂O)_m-、-(C_1-6 alkyl) N (R) C (O) (C _1-6 alkyl) -and- (CH ₂CH₂O)_m(C_1-6 alkyl) N (R) C (O) (C _1-6 alkyl) -, where m is 1-4. In some embodiments, -T-is selected from the group consisting of- (CH ₂CH₂O)_m-、-(C_3-5 alkyl) N (R) C (O) (C _2-4 alkyl) -and- (CH ₂CH₂O)_m(C_3-5 alkyl) N (R) C (O) (C _2- ₄ alkyl) -, where m is 2-3. In some embodiments, -T-is selected from:

In some embodiments, the detectable moiety R ^t is attached to the compound of formula I by click chemistry (CLICK CHEMISTRY). In some embodiments, the compound of formula I is attached to-T-R ^t by 1, 3-cycloaddition of azide to 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, angel. 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 reserve inhibitor (CLICK READY inhibitor). In some such embodiments, the click back-up inhibitor of formula IV reacts with the click back-up-T-R ^t moiety. As used herein, "click ready" refers to an azide or alkyne containing moiety for click chemistry reactions. In some embodiments, clicking on the alternate inhibitor moiety comprises azide. In certain embodiments, the click back-up-T-R ^t portion contains strained cyclooctyne for a copper-free click chemistry (e.g., using the methods described in Baskin et al, proc. Natl. Acad. Sci. USA,2007,104,16793-16797).

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

Preparation of Compounds of formula I

The compounds of formula I can be prepared as shown in schemes 1-4 below.

Route 1

Wherein R ¹ and R ⁴ are as defined in any of the embodiments above.

As shown in scheme 1, the compound of formula a is reacted with lawenson's reagent in a suitable organic solvent at elevated temperature to give the compound of formula B. The organic solvent includes, but is not limited to, toluene, xylene, preferably toluene. The elevated temperature may be, for example, 80-140 ℃, in particular 90-120 ℃, or 100-110 ℃.

Route 2

Wherein R ³ is as defined in any embodiment above.

As shown in scheme 2, an amine of formula C is reacted with thiophosgene D in the presence of a suitable base in a suitable organic solvent at a suitable temperature to provide an isocyanate of formula E. The suitable temperature may be, for example, about 0 ℃. The base is, for example, an organic amine, in particular an alkylamine, such as triethylamine. The organic solvent includes Dichloromethane (DCM). The reaction may be carried out under an inert atmosphere, for example under nitrogen.

Route 3

As shown in scheme 3, the compound of formula B is reacted with isocyanate E in the presence of a suitable base at a suitable temperature in a suitable organic solvent to provide a compound of formula I' (i.e., a compound of formula I wherein R ² is H). The base may be an inorganic base, in particular an alkali metal hydroxide, such as NaOH or KOH. The organic solvent includes N, N-Dimethylformamide (DMF). The suitable temperature may be, for example, room temperature (20-35 ℃).

Route 4

As shown in scheme 4, reacting compound I' with an anhydride of formula F in the presence of a suitable base in a suitable organic solvent at a suitable temperature gives a compound of formula I wherein R ² is C _1-6 alkyl. The base may be an inorganic base, in particular an alkali metal carbonate, such as sodium bicarbonate, potassium bicarbonate. The organic solvent includes, for example, DCM. The suitable temperature may be, for example, about 0 ℃.

Composition and method for producing the same

In some embodiments, the compounds of formula I may be provided, for example, in the form of a composition in combination (e.g., in admixture) with one or more other components.

In some embodiments, the present disclosure provides compositions comprising and/or delivering a compound of formula I, e.g., when contacted with or otherwise applied to a system or environment, e.g., the system or environment may include SARM1NAD enzymatic activity; in some embodiments, administration of the composition to the system or environment can achieve inhibition of SARM1 activity described herein.

In some embodiments, the compositions described herein may be pharmaceutical compositions, wherein the pharmaceutical compositions comprise an active agent and one or more pharmaceutically acceptable carriers. In some such embodiments, the pharmaceutical composition comprises and/or delivers the compound of formula I to a related system or environment described herein (e.g., to an individual in need thereof).

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

The present disclosure also provides (pharmaceutical) compositions comprising a compound of formula I and a pharmaceutically acceptable carrier, adjuvant or vehicle. The amount of the compound in the composition is such that the axonal degeneration in the biological sample or patient is effectively measurably inhibited. In certain embodiments, the compounds or compositions are formulated for administration to a patient in need of such compositions. According to the methods of the present disclosure, the compounds and compositions may be administered using any amount and any route of administration effective to treat or reduce the severity of any disease or disorder described herein. The compounds are preferably formulated in unit dosage form to facilitate administration and uniformity of dosage. The expression "unit dosage form" as used herein refers to physically discrete units of medicament suitable for the patient to be treated. However, it will be appreciated that the total daily amount of the compounds and compositions will be determined by the attending physician within the scope of sound medical judgment. The specific effective dosage level for any particular patient or organism will vary from individual to individual, depending on a variety of factors, including the condition being treated and the severity of the condition; the activity of the particular compound employed; the particular composition employed and the route of administration thereof; the species, age, weight, sex and diet of the patient; the general condition of the individual; the time of application; excretion rate of the specific compound employed; duration of treatment; drugs used in combination with or simultaneously with the particular compound employed, and the like.

The compositions provided by the present disclosure may be administered orally, parenterally, by inhalation or nasal spray, topically (e.g., by powder, ointment, or drops), rectally, bucally, intravaginally, intraperitoneally, intracisternally, or by an implanted reservoir, depending on the severity of the condition being treated. Preferably, the composition is administered orally, intraperitoneally, or intravenously. In certain embodiments, the compounds are administered orally or parenterally at a dosage level of about 0.01mg/kg to about 50mg/kg of body weight of the individual, one or more times per day, to achieve the desired therapeutic effect.

The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. The sterile injectable form of the composition may be an aqueous or oleaginous 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, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tweens (Tweens), spandex, and other emulsifying agents or bioavailability enhancers, are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms, and may also be used for formulation purposes.

The injectable formulations may be sterilized, for example, by filtration through bacterial-retaining filters, or by incorporating sterilizing agents in the form of sterile solid compositions which may be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

In order to prolong the effect of the compounds, it is often desirable to slow down the absorption of the compounds by subcutaneous or intramuscular injection. This can be achieved by using liquid suspensions of crystalline or amorphous materials that are poorly water soluble. The absorption rate of a compound then depends on its dissolution rate, which in turn may depend on crystal size and crystal morphology. Or by dissolving or suspending the compound in an oily vehicle to achieve delayed absorption of the parenterally administered compound form. The injectable depot forms are made by forming a matrix of microcapsules of the compound in a biodegradable polymer such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the particular polymer employed, the release rate of the compound may be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

The pharmaceutically acceptable compositions provided by the present disclosure may be administered orally in any orally acceptable dosage form, including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In such solid dosage forms, the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also contain, in addition to inert diluents, additional substances such as lubricants and other tabletting aids, such as magnesium stearate and microcrystalline cellulose, as is usual. When an aqueous suspension for oral administration is desired, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweeteners, flavoring agents or coloring agents may also be added.

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 dicalcium phosphate, and/or a) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and silicic acid; b) Binders such as carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; c) Humectants, such as glycerol; d) Disintegrants, 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 forms may also comprise buffering agents. The active compound may also be in microencapsulated form with one or more excipients as described above.

Solid compositions of a similar type may also be used as fillers for soft and hard filled gelatin capsules using lactose or milk sugar as well as excipients such as 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., buffers) and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and may also be compositions which release the active ingredient(s) only or preferably in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that may be used include polymeric substances and waxes.

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

Or the pharmaceutically acceptable compositions provided by the present disclosure may be administered in the form of suppositories for rectal or vaginal administration. These may be prepared by mixing a compound of the present disclosure with a suitable non-irritating excipient or carrier which is solid at room temperature but liquid at body temperature (e.g. rectal or vaginal temperatures) and therefore will melt in the rectum or vaginal cavity to release the active compound. Such materials include cocoa butter, suppository waxes (e.g., beeswax) and polyethylene glycols.

The pharmaceutically acceptable compositions provided by the present disclosure may also be administered topically, particularly when the therapeutic target includes an area or organ readily accessible by topical administration, including diseases of the eye, skin, or lower intestinal tract. Topical administration to the lower intestinal tract may be effected in rectal suppository formulations (see above) or in suitable enema formulations.

Dosage forms for topical or transdermal administration of the compounds provided by the present disclosure include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier and any required preservatives or buffers as may be required. Ophthalmic formulations, ear drops, and eye drops are also contemplated as falling within the scope of the present disclosure. Furthermore, the present disclosure also contemplates the use of transdermal patches, which have the additional advantage of controllably delivering the compound into the body. Such dosage forms may be prepared by dissolving or dispersing the compound in a suitable medium. Absorption enhancers may also be used to increase the flux of a compound across the skin. The rate may be controlled by providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

For topical application, the pharmaceutically acceptable compositions provided by the present disclosure may be formulated as a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical application of the compounds of the present disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the provided pharmaceutically acceptable compositions may 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, cetostearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutically acceptable compositions provided by the present disclosure may be formulated as micronized suspensions in isotonic, pH adjusted sterile physiological saline, or preferably as solutions in isotonic, pH adjusted sterile physiological saline, with or without a preservative such as benzalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutically acceptable composition may be formulated in an ointment such as petrolatum.

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

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

Identification and/or characterization of compounds and/or compositions

The present disclosure provides, among other things, various techniques for identifying and/or characterizing compounds and/or compositions described herein. For example, the present disclosure provides various assays for assessing SARM1 inhibition activity, particularly for assessing SARM1 inhibition activity.

In some embodiments, the performance of one or more compounds or compositions of interest in the assays described herein is compared to a performance of an appropriate reference. For example, in some embodiments, the reference may be the absence of a related compound or composition. Alternatively or additionally, in some embodiments, the reference may be the presence of an alternative compound or composition, e.g., having a known property in a relevant assay (e.g., as a positive control or a negative control, as understood in the art). In some embodiments, the reference may be an alternative but comparable set of conditions (e.g., temperature, pH, salt concentration, etc.). In some embodiments, the reference can be a property of the compound or composition relative to the SARM1 variant.

Further alternatively or additionally, in some embodiments, in an assay described herein, the properties of one or more compounds or compositions of interest may be assessed in the presence of an appropriate reference compound or composition, e.g., to determine the ability of the compound or composition to compete with a reference.

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

Some exemplary assays that may be used in the practice of the present disclosure are illustrated in the following examples. Those of skill in the art having read the present disclosure will appreciate that useful or related 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 to those discussed further below.

In some embodiments, the compounds and/or compositions may be determined based on, and/or characterized by, one or more activities or characteristics, such as, for example: promoting axon integrity, cytoskeletal stability and/or neuronal survival. In some embodiments, provided SARM1 inhibitors inhibit catabolism of nad+ by SARM 1. In some embodiments, provided SARM1 inhibitors slow the rate of nad+ catabolism.

In some embodiments, provided SARM1 inhibitors reduce or inhibit the binding of SARM1 to nad+. In some embodiments, provided SARM1 inhibitors bind to SARM1 within a pocket comprising one or more catalytic residues (e.g., a catalytic cleft of SARM 1). Examples of such catalytic residues include glutamic acid at position 642 (E642).

In some embodiments, provided SARM1 inhibitors disrupt and/or prevent multimerization of the TIR1 domain of SARM 1. In some embodiments, provided SARM1 inhibitors disrupt multimerization of SAM domains. In some embodiments, provided SARM1 inhibitors disrupt an axon signaling cascade that leads to nad+ depletion.

In some embodiments, the present disclosure provides assays useful for identifying and/or characterizing one or more activities and/or characteristics of a compound and/or composition of interest. For example, in some embodiments, the present disclosure provides in vitro, cellular, and/or in vivo systems for assessing one or more of such activities and/or characteristics.

SARM1 Activity assay

In some embodiments, a method of identifying an inhibitor of SARM1 comprises: a) Providing a mixture comprising i) a mutant or fragment of SARM1, ii) nad+ and iii) a candidate inhibitor, wherein the mutant or fragment has constitutive activity; b) Incubating the mixture; c) Quantifying nad+ in the mixture after incubation; and d) identifying the candidate inhibitor compound as an inhibitor if the amount of nad+ is greater than the amount of the control mixture that does not contain the candidate inhibitor.

In some embodiments, methods of identifying an inhibitor of SARM1 are provided comprising: 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) Quantification of nad+ and ADPR (or cADPR) in the mixture after incubation; d) Determining the molar ratio of NAD+: ADPR (or cADPR); and e) identifying a candidate inhibitor compound as an inhibitor if the molar ratio is greater than the molar ratio of a control mixture that does not contain the candidate inhibitor.

In some embodiments, methods of identifying an inhibitor of SARM1 are provided comprising: 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 the 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 the control.

SARM1 binding assay

In some embodiments, the efficacy of the provided SARM1 inhibitors can be determined from an assay described in WO 2018/057989, for example, publication No. 3/29, 2018, which is incorporated herein by reference in its entirety. In some embodiments, provided SARM1 inhibitors may be applied to solutions containing SARM1 or fragments thereof. In some embodiments, provided SARM1 inhibitors may be applied to in vitro systems. In some embodiments, provided SARM1 inhibitors may be applied to in vivo systems. In some embodiments, provided SARM1 inhibitors may be applied to a patient. In some embodiments, the SARM1 inhibitor may be admixed with SARM1 or a fragment thereof which has been labeled with an epitope tag. In some embodiments, the amount of bound SARM1 inhibitor may be compared to the amount of unbound SARM1 inhibitor to derive an affinity for the SARM1 inhibitor.

In some embodiments, the mutant or fragment of SARM1 is a fragment of SAM-TIR having constitutive activity. SARM1 fragments having constitutive activity include, for example, but are not limited to, SARM1 lacking a self-inhibiting domain; at least one point mutation of SARM1 that inactivates the self-inhibiting domain; a SARM1 fragment containing a TIR domain; or a SARM1 fragment consisting of SAM and TIR domains. In some embodiments, the SARM1 polypeptide may include one or more additional amino acid sequences that may serve as tags, such as His tags, streptavidin tags, or combinations thereof. In some embodiments, the SARM1 polypeptide may comprise a tag at the amino terminus, the carboxy terminus, or a combination thereof. In some embodiments, an epitope-tagged SARM1 or fragment thereof is useful for measuring the binding efficacy of a provided SARM1 inhibitor.

Purification of SARM1-TIR domains

In some embodiments, the SARM1-TIR domain may be engineered with various protein tags or epitope tags that may be used, for example, for purification. In some embodiments, the disclosure also provides an NRK1-HEK293T cell line comprising HEK293T cells transformed with nicotinamide riboside kinase 1 (NRK 1). In some embodiments, HEK293T cells are transformed or transfected with a DNA sequence encoding nicotinamide riboside kinase 1 (NRK 1). In some embodiments, the DNA encoding NRK1 may be genomic or cDNA. In some embodiments, HEK293T cells are stably or transiently transfected with DNA encoding NRK1 from outside 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 (e.g., promoters, enhancers, or combinations thereof). In some embodiments, the combination of DNA sequences encoding NRK1 and 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 FCIV expression vectors. 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, a human, a mouse, a zebra fish, or a drosophila. In some configurations, the NRK1DNA is human NRK1DNA.

Application and use

The present disclosure provides various uses and applications of the compounds and/or compositions described herein, for example, in accordance with the activity and/or characteristics of such compounds and/or compositions as described herein. In some embodiments, such uses may include therapeutic and/or diagnostic uses. Or in some embodiments such uses may include research, production, and/or other technical uses.

In one aspect, the present disclosure provides methods comprising administering one or more compounds of formula I to an individual, e.g., to treat, prevent, or reduce the risk of suffering from one or more diseases, disorders, or conditions characterized by axonal degeneration. In some such embodiments, the compound of formula I is a SARM1 inhibitor.

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

Inhibition of enzymes in biological samples is useful for a variety of purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, bioassays, gene expression studies, and biological target identification.

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. In some embodiments, one or more compounds and/or compositions described herein may be used, for example, in methods of inhibiting neuronal degeneration from an individual. In some embodiments, one or more compounds and/or compositions described herein can be used to inhibit the degeneration of neurons or a portion thereof in vitro cultured. In some embodiments, one or more compounds and/or compositions described herein may be used as stabilizers to promote neuronal survival in vitro.

In some embodiments, the provided compounds and/or compositions inhibit NAD enzymatic activity of SARM 1. Alternatively or additionally, in some embodiments, the provided compounds alleviate one or more properties of neurodegeneration. In some embodiments, the present disclosure provides methods of treating a neurodegenerative disease, disorder, or condition associated with axonal degeneration.

In some embodiments, one or more compounds and/or compositions described herein may be used, for example, in medical practice. In some embodiments, one or more compounds and/or compositions described herein can be used, for example, to treat, prevent, or ameliorate axonal degeneration (e.g., one or more features or characteristics thereof). In some embodiments, one or more compounds and/or compositions described herein can be used, for example, to inhibit axonal degeneration, including axonal degeneration resulting from nad+ reduction or depletion. In some embodiments, one or more compounds and/or compositions described herein may be used, for example, to prevent axonal degeneration distal to an axonal injury.

In some embodiments, one or more compounds and/or compositions described herein may be used in methods of inhibiting degeneration of a peripheral nervous system neuron or a portion thereof, for example. In some embodiments, one or more compounds and/or compositions described herein may be used in methods of, for example, inhibiting or preventing degeneration of the central nervous system (neurons) or a portion thereof. In some embodiments, one or more compounds or compositions described herein are characterized by reducing one or more symptoms or features of neurodegeneration when administered to a population of individuals. For example, in some embodiments, the associated symptom or feature may be selected from the group consisting of the degree, rate, and/or timing of neuronal damage.

In certain embodiments, the present disclosure provides compounds according to the present disclosure for use, for example, as an analytical tool, as a probe in a bioassay, or as a therapeutic agent. The compounds provided by the present disclosure are also useful for studying SARM1 activity in biological and pathological phenomena, as well as for comparative evaluation of novel inhibitors of SARM1 activity in vitro or in vivo. In certain embodiments, the present disclosure provides assays for identifying and/or characterizing 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 determine SARM1 activity. For example, in some embodiments, the provided assays may utilize SAM-TIR of the auto-frustrated domain, e.g., the absence of the SARM 1N-terminus, and/or one or more tagged versions of the TIR domain.

In some embodiments, one or more compounds and/or compositions described herein may be used, for example, in methods of inhibiting neuronal degeneration from an individual. In some embodiments, one or more compounds and/or compositions described herein can be used to inhibit the degeneration of neurons or a portion thereof in vitro cultured. In some embodiments, one or more compounds and/or compositions described herein may be used as stabilizers to promote neuronal survival in vitro.

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

In some embodiments, one or more compounds and/or compositions described herein can affect a detectable change in the level of one or more neurodegeneration associated proteins in an individual. Such proteins include, but are not limited to, albumin, amyloid-beta (aβ) 38, aβ40, aβ42, glial Fibrillary Acidic Protein (GFAP), cardiac fatty acid binding protein (hFABP), monocyte Chemotactic Protein (MCP) -1, neuroparticulate protein, neuron-specific enolase (NSE), soluble amyloid precursor protein (sAPP) alpha, sappβ, soluble trigger receptor expressed on myeloid cells (sTREM) 2, phosphorylated tau, and/or total tau (total-tua). 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.

Diseases, disorders and conditions

In some embodiments, the compounds and/or compositions described herein may be administered to an individual suffering from one or more diseases, disorders, or conditions.

In some embodiments, the disease, disorder, or condition is acute. In some embodiments, the disease, disorder, or condition is chronic.

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

In some embodiments, the disease, disorder or condition is or comprises 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 disease, disorder or condition is or comprises chronic injury to the central nervous system, such as injury to the spinal cord, traumatic brain injury, and/or traumatic axonal injury. In some embodiments, the disease, disorder, or condition is or comprises chronic traumatic brain injury (CTE).

In some embodiments, the disease, disorder or 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.

In some embodiments, the disease, disorder, or condition is acute peripheral neuropathy. Chemotherapy-induced peripheral neuropathy (CIPN) is an example of an acute peripheral neuropathy. CIPN may be associated with a variety of agents such as, but not limited to, thalidomide (thalidomide), epothilone (epothilone) (e.g., ixabepilone (ixabepilone)), taxane (e.g., paclitaxel (paclitaxel) and docetaxel), vinca alkaloid (vinca alloid) (e.g., vinblastine (vinblastine), vinorelbine (vinorelbine), vincristine (vincristine) and vindesine (vindesine)), proteasome inhibitors (e.g., bortezomib (bortezomib)), platinum-based agents (e.g., cisplatin (cisplatin), oxaliplatin (oxaliplatin), and carboplatin (carboplatin)).

In some embodiments, the disease, disorder, or condition is a chronic condition affecting the peripheral nervous system, such as diabetic neuropathy, HIV neuropathy, charcot-marie-tooth's disease (Charcot Marie Tooth disease), or amyotrophic lateral sclerosis.

In some embodiments, the disease, disorder, or condition is an acute condition affecting the optic nerve, such as Acute Optic Neuropathy (AON) or acute angle closure glaucoma.

In some embodiments, the disease, disorder or condition is a chronic condition affecting the optic nerve, such as leber's congenital amaurosis, leber's hereditary optic neuropathy, primary open angle glaucoma, and autosomal dominant optic atrophy.

In some embodiments, one or more compounds and/or compositions described herein may be used, for example, to treat one or more neurodegenerative diseases, disorders, or conditions selected from neuropathy or axonal lesions. In some embodiments, one or more compounds and/or compositions described herein may be used, for example, to treat neuropathy or axonal lesions associated with axonal degeneration. In some embodiments, the neuropathy associated with axonal degeneration is hereditary or congenital neuropathy or axonal lesions. In some embodiments, the neuropathy associated with axonal degeneration is caused by neogenesis or somatic mutation. In some embodiments, the neuropathy associated with axonal degeneration is selected from the list contained herein. In some embodiments, the neuropathy or axonal lesions are 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, demyelinating diseases, ischemia or stroke, chemical injury, thermal injury, and AIDS.

In some embodiments, one or more compounds or compositions described herein are characterized by reducing one or more symptoms or features of neurodegeneration when administered to a population of individuals. For example, in some embodiments, the associated symptom or feature may be selected from the group consisting of the degree, rate, and/or timing of neuronal damage. In some embodiments, the neuronal disruption may be or comprise axonal degeneration, loss of synapse, loss of dendrite, loss of synaptic density, loss of dendrite arbor, loss of axonal branching, loss of neuronal density, loss of myelination, loss of neuronal cell mass, loss of synaptic enhancement, loss of action potential enhancement, 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 capacity, loss of axonal potential transmission, neuronal hyperexcitability and/or neuronal hypoexcitability. In some embodiments, the neuronal disruption is characterized by the inability to maintain an appropriate resting neuronal membrane potential. In some embodiments, the neuronal destruction is characterized by the presence of inclusion bodies, plaques, and/or neurofibrillary tangles. In some embodiments, the neuronal destruction is characterized by the presence of stress particles. In some embodiments, the neuronal disruption is characterized by intracellular activation of one or more members of the cysteine-aspartic protease (Caspase) family. In some embodiments, the neuronal destruction is characterized by the neurons undergoing programmed cell death (e.g., apoptosis, coking, iron death, and/or necrosis) and/or inflammation.

In some embodiments, the neurodegenerative or nervous system disease, disorder or condition is associated with axonal degeneration, axonal injury, axonal lesions, demyelinating diseases, central pontine myelinolysis, nerve injury diseases or disorders, metabolic diseases, mitochondrial diseases, metabolic axonal degeneration, axonal injury caused by leukoencephalopathy or leukodystrophy. In some embodiments, the neurodegenerative or neurological disease, disorder or condition is selected from: spinal cord injury, stroke, multiple sclerosis, progressive multifocal leukoencephalopathy, reduced congenital myelination, encephalomyelitis, acute disseminated encephalomyelitis, central pontine myelinolysis, hyponatremia, hypoxic demyelination, ischemic demyelination, adrenoleukodystrophy, alexander's disease, niemann-pick disease (Niemann-PICK DISEASE), paMey disease (PelizaeusMerzbacher disease), Periventricular leukomalacia, globoid leukodystrophy (Krabbe's disease), schlemm's 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, heller syndrome (HurlerSyndrome), traumatic brain injury, Post-radiation injury, neurological complications of chemotherapy (chemotherapy-induced neuropathy; CIPN), neuropathy, acute ischemic optic neuropathy, vitamin B ₁₂ deficiency, shan Weisheng-E deficiency syndrome, barker's syndrome (Bassen-Kornzweig syndrome), glaucoma, leber's hereditary optic atrophy (neuropathy), leber's congenital amaurosis, neuromyelitis optica, metachromatic leukodystrophy, acute hemorrhagic leukoencephalopathy, trigeminal neuralgia, bell's palsy (Bell's palsy), and, Cerebral ischemia, multiple system atrophy, traumatic glaucoma, tropical spastic paraplegia human T lymphotropic virus 1 (HTLV-1) -associated myelopathy, west Nile virus encephalopathy (west Nile virusencephalopathy), rake's virus encephalitis (La Crosse virus encephalitis), bunyavirus encephalitis (Bunyavirus encephalitis), pediatric viral encephalitis, essential tremor, summer-horse-figure three disease, motor neuron disease, spinal Muscular Atrophy (SMA), Hereditary Sensory and Autonomic Neuropathy (HSAN), adrenospinal neuropathy, progressive Supranuclear Palsy (PSP), friedrich's ataxia), hereditary ataxia, noise-induced hearing loss, congenital hearing loss, lewy body dementia (Lewy Body Dementia), frontotemporal dementia, amyloidosis, diabetic neuropathy, HIV neuropathy, enteric neuropathy and axonopathy, guillain-Barre syndrome (Guillain-Barre syndrome), severe Acute Motor Axonopathy (AMAN), Creutzfeldt-Jakob disease, transmissible spongiform encephalopathy, spinocerebellar ataxia, preeclampsia, hereditary spastic paraplegia, spastic paraplegia familial, french colonisation disease (FRENCH SETTLEMENT DISEASE), st-Lo disease (Strumpell-Lorrain disease) and nonalcoholic steatohepatitis (NASH).

In some embodiments, the present disclosure provides inhibitors of SARM1 activity for the treatment of neurodegenerative or nervous system diseases or disorders involving axonal degeneration or axonal lesions. The present disclosure also provides methods of using inhibitors of SARM1 activity to treat, prevent, or ameliorate axonal degeneration, axonal lesions, and neurodegenerative or nervous system diseases, disorders, or conditions involving axonal degeneration.

In some embodiments, the present disclosure provides methods of treating a neurodegenerative or neurological disease, disorder or condition associated with axonal degeneration, axonal injury, axonal lesions, demyelinating diseases, pontine myelinolysis, nerve injury diseases or disorders, metabolic diseases, mitochondrial diseases, metabolic axonal degeneration, axonal injury caused by leukoencephalopathy or leukodystrophy.

In some embodiments, neuropathy and axonal lesions include any disease, disorder, or condition involving neurons and/or supporting cells, such as glial cells, muscle cells, or fibroblasts, particularly those involving axonal injury. Axonal injury may be caused by traumatic injury or by non-mechanical injury caused by a disease, disorder or condition or exposure to toxic molecules or drugs. The result of such damage may be degeneration or dysfunction of axons and loss of functional neuronal activity. The diseases, disorders and conditions that result or are associated with such axonal damage are one of many neurological diseases, disorders and conditions. Such neuropathy may include peripheral neuropathy, central neuropathy, and combinations thereof. Furthermore, peripheral neurological manifestations may result mainly from diseases focused on the central nervous system, whereas central nervous system manifestations may result substantially from peripheral or systemic diseases.

In some embodiments, peripheral neuropathy may involve damage to peripheral nerves, and/or may be caused by disease of the nerves or due to systemic disease. Some such diseases may include diabetes, uremia, infectious diseases such as AID 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 neurodegeneration is caused by traumatic (mechanical) injury to the nerve as well as chemical or thermal injury to the nerve. Such conditions that damage peripheral nerves include compression or clamping injuries such as glaucoma, carpal tunnel syndrome, direct trauma, penetrating injury, contusion, bone fracture or bone dislocation; pressure related to the superficial nerve (ulna, radius or fibular nerve), which may be due to prolonged use of the crutch or prolonged residence time in one location, or due to a tumor; internal hemorrhage of nerve; ischemia; exposure to cold or radiation or certain drugs or toxic substances, such as herbicides or pesticides. In particular, the nerve damage may be due to chemical damage 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 sensation such as burning, itching, stinging or tingling) and pain in the arms, hands, legs and/or feet. In some embodiments, the neuropathy is associated with mitochondrial dysfunction. Such neuropathy may exhibit reduced energy levels, i.e., reduced NAD and ATP levels.

In some embodiments, the peripheral neuropathy is a metabolic and endocrine neuropathy, which includes a broad range of peripheral neurological 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 metabolism disorders, nutritional/vitamin deficiency disorders, and mitochondrial disorders, among others. A common feature of these diseases is involvement of the peripheral nerves due to structural or functional changes in myelin and axons caused by metabolic pathway disorders.

In some embodiments, the neuropathy comprises optic neuropathy, such as glaucoma; retinal ganglion degeneration, such as retinal ganglion degeneration associated with retinitis pigmentosa and epiretinal neuropathy; optical neuritis and/or degeneration, including those associated with multiple sclerosis; traumatic injury to the optic nerve, which may include, for example, injury during tumor resection; hereditary optic neuropathy, such as Kaposi's disease (Kjer's disease) and Libert hereditary optic neuropathy; ischemic optic neuropathy, such as optic neuropathy secondary to giant cell arteritis; metabolic optic neuropathy, such as neurodegenerative diseases, including the aforementioned leber's neuropathy, nutritional deficiency, such as vitamin B12 or folate deficiency, and poisoning, such as due to ethambutol or cyanide; neuropathy caused by adverse drug reactions and neuropathy caused by vitamin deficiency. Ischemic optic neuropathy also includes non-arterial anterior ischemic optic neuropathy.

In some embodiments, the neurodegenerative disease associated with neuropathy or axonal lesions in the central nervous system comprises a variety of diseases. Such diseases include diseases involving progressive dementia, such as Alzheimer's disease, senile dementia, pick's disease and Huntington's disease; central nervous system diseases affecting muscle function, such as parkinson's disease, motor neuron disease, and progressive ataxia, such as amyotrophic lateral sclerosis; demyelinating diseases, such as multiple sclerosis; viral encephalitis, such as that caused by enteroviruses, arboviruses, and herpes simplex viruses; prion diseases. Mechanical injuries, such as glaucoma or traumatic injuries to the head and spine, can also cause nerve damage and degeneration of the brain and spinal cord. In addition, ischemia and stroke, as well as conditions such as nutritional deficiencies and chemotoxicity (e.g., chemotherapeutic agents), can also cause central nervous system neuropathy.

In some embodiments, the present disclosure provides a method of treating a neuropathy or axonal disorder associated with axonal degeneration. In some such embodiments, the neuropathy or axonal disorder associated with axonal degeneration may be any of a variety of neuropathy or axonal lesions, such as those that are hereditary or congenital or are associated with parkinson's disease, alzheimer's disease, herpes infections, diabetes, amyotrophic lateral sclerosis, demyelinating diseases, ischemia or stroke, chemical injury, thermal injury, and AIDS. Furthermore, the above-mentioned neurodegenerative diseases, as well as a subset of the above-mentioned diseases, may also be treated with the methods of the present disclosure. Such a subset of diseases may include parkinson's disease or non-parkinson's disease, or alzheimer's disease.

Individual body

In some embodiments, the compounds and/or compositions described herein are administered to an individual suffering from or susceptible to a disease, disorder, or condition described herein; in some embodiments, such a disease, disorder, or condition is characterized by axonal degeneration, as one of the diseases, disorders, or conditions mentioned herein.

In some embodiments, an individual to whom a compound or composition described herein is administered exhibits one or more signs or symptoms associated with axonal degeneration; in some embodiments, the individual does not exhibit any signs or symptoms of neurodegeneration.

In some embodiments, provided methods comprise administering a compound of formula I to a patient in need thereof. In some such embodiments, the patient is at risk of suffering from a disease, disorder, or condition characterized by axonal degeneration. In some embodiments, the patient has a disease, disorder, or condition characterized by axonal degeneration. In some embodiments, the patient has been diagnosed with a disease, disorder, or condition characterized by axonal degeneration.

In some embodiments, provided methods comprise administering to a population of patients in need thereof a composition described herein. In some embodiments, the population is from those individuals involved in activities with a high likelihood of traumatic neuronal injury. In some embodiments, the population is from athletes engaged in contact sports or other high risk activities.

In some embodiments, the subject is at risk of suffering from a disease, disorder, or condition characterized by axonal degeneration. In some embodiments, the individual is identified as being at risk for axonal degeneration, e.g., based on the genotype of the individual, diagnosis of a disease, disorder or condition associated with axonal degeneration, and/or exposure to agents and/or conditions that induce axonal degeneration.

In some embodiments, the patient is at risk of suffering from 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. 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 neurodegenerative high-incidence population. In some embodiments, the patient has a repeated amplification of a hexanucleotide in chromosome 9 open reading frame 72. In some embodiments, the patient has one or more copies of an ApoE4 allele.

In some embodiments, an individual to whom a compound or composition described herein is administered may be or comprise an individual suffering from or susceptible to a neurodegenerative disease, disorder or condition. In some embodiments, the neurodegenerative disease, disorder, or condition may be or comprise traumatic neuronal injury. In some embodiments, the traumatic neuronal injury is a blunt force wound, a closed head injury, an open head injury, a penetrating injury to the brain cavity or a innervated area of the body, exposed to oscillating and/or explosive forces. In some embodiments, the traumatic neuronal injury is a force that causes axonal deformation, stretching, squeezing, or shearing.

In some embodiments, the individual engages in an activity identified as a risk factor for neuronal degeneration, e.g., an individual engaged in a occupational with a high probability of contact exercise or traumatic neuronal injury.

For example, the individual may be a patient who is receiving or is 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-based drugs (e.g., cisplatin, oxaliplatin, and carboplatin).

In some embodiments, provided methods comprise administering a composition described herein to a patient or patient population based on the presence or absence of one or more biomarkers. In some embodiments, the provided methods further comprise monitoring the level of the biomarker in the patient or patient population and adjusting the dosing regimen accordingly.

Administration of drugs

It will be appreciated by those of skill in the art that in some embodiments, the precise amount of a particular compound included in and/or delivered by administration in a pharmaceutical composition or regimen described herein may be selected by a healthcare practitioner and may vary from individual to individual, e.g., after considering one or more of the individual's species, age and general condition, and/or the nature of the particular compound or composition, its mode of administration, etc. Or in some embodiments, the amount of a particular compound included in a pharmaceutical composition or regimen as described herein and/or delivered by administration can be normalized in a relevant patient population (e.g., all patients of a particular age or disease stage or expressing a particular biomarker, etc.).

The compounds or compositions provided by the present disclosure are preferably formulated in unit dosage form to facilitate administration and uniformity of dosage. The expression "unit dosage form" as used herein refers to physically discrete units of medicament suitable for the patient to be treated. However, it will be appreciated 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 sound medical judgment. The specific effective dosage level for any particular patient or organism will depend on a variety of factors, including the condition being treated and the severity of the condition; clinical condition of individual patient; the cause of the disorder; the activity of the particular compound employed; the specific composition employed; age, weight, general health, sex and diet of the patient; the time of administration of the particular compound employed, the site of delivery of the agent, the route of administration and the rate of excretion; duration of treatment; drugs used in combination or concurrently with the particular compound employed, and the like as is 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 in order to prevent or treat an undesired disease or disorder, such as neurodegeneration or traumatic nerve injury.

The pharmaceutically acceptable compositions of the present disclosure may be administered to humans and other animals orally, rectally, intravenously, parenterally, intracisternally, intravaginally, intraperitoneally, topically (e.g., as powders, ointments or drops), bucally, 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 administered in a single dose per day or in divided doses of from twice to six times per day, or in sustained release form. This dosing regimen may be adjusted to provide the optimal therapeutic response. The compounds may be administered on a regimen of 1 to 4 times per day, preferably once or twice per day.

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

In some embodiments, the pharmaceutically acceptable compositions of the present disclosure may also be administered topically, particularly when the therapeutic target includes an area or organ readily accessible by topical administration, including diseases of the eye, skin, or lower intestinal tract. Topical formulations suitable for each of these areas or organs are readily prepared.

Most preferably, the pharmaceutically acceptable compositions of the present disclosure are formulated for oral administration. Such formulations may 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.

Those additional agents may be administered as part of a multi-dose regimen, separately from the provided compounds or compositions thereof. Or those agents may be part of a single dosage form, mixed together with the provided compounds in a single composition. If administered as part of a multi-dose regimen, the two active agents may be provided simultaneously, sequentially or within a period of time of each other (typically within five hours of each other).

It will also be appreciated that the particular dosage and treatment regimen of any particular patient may depend on a variety of factors including the activity of the particular compound employed, the 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 a compound of the present disclosure in a composition will also depend on the particular compound in the composition.

In some embodiments, the SARM1 inhibitors described herein can be utilized in combination with one or more other therapies to treat a related disease, disorder, or condition. In some embodiments, the dosage of the SARM1 inhibitor is altered when using the combination therapy as compared to when administered as monotherapy; alternatively or additionally, in some embodiments, the therapy administered in combination with SARM1 inhibition described herein is administered according to a regimen or treatment regimen other than the regimen (region) or treatment regimen (protocol) 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, and the provided compound may act synergistically. In some embodiments, one or both therapies used in the combination regimen are administered at a lower level or less frequently than when used as monotherapy.

In some embodiments, the compounds and/or compositions described herein are administered 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-based agents, retinoids, vinca alkaloids, and derivatives. In some embodiments, the compounds and/or compositions described herein are administered in combination with a PARP inhibitor.

Advantageous effects of the invention

The inventors surprisingly found that the compounds provided by the present disclosure, obtained by replacing the oxo (= O) group on 2, 3-dihydroisothiazole with the thio (= S) group, have significantly improved SARM1 inhibitory activity compared to the corresponding compounds disclosed in WO2019236890A1 (e.g. positive controls 1 and 2 determined in the biological examples below). The compounds of the present disclosure also have a variety of excellent properties, such as good physicochemical properties (e.g., solubility, physical and/or chemical stability), good pharmacokinetic properties (e.g., good drug exposure and good oral absorption effects), good safety (lower toxicity and/or fewer side effects, wider therapeutic window).

Examples

Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.

NMR was performed using Bruker AVANCE-400 and Bruker AVANCE-500 nuclear magnetic instruments, and the measurement solvents included deuterated dimethyl sulfoxide (DMSO-d 6), deuterated acetone (CD ₃COCD₃), deuterated chloroform (CDCl ₃), deuterated methanol (CD ₃ OD), etc., with the internal standard being Tetramethylsilane (TMS), and chemical shifts being measured in parts per million (ppm).

Liquid chromatography (LC-MS) was performed using an Agilent 1260 mass spectrometer. HPLC was determined using an Agilent1100 high pressure chromatograph (Microsorb microns C18.times.3.0 mm column).

The thin layer chromatography silica gel plate is Qingdao GF254 silica gel plate, TLC is 0.15-0.20mm, and the preparation thin layer chromatography is 0.4-0.5 mm. Column chromatography generally uses Qingdao silica gel 200-300 mesh silica gel as carrier.

Abbreviations in the context of the present disclosure have the following meanings:

Example 1: synthesis of 5- ((2-chlorophenyl) amino) -3-mercaptoisothiazole-4-carbonitrile (5)

1-Chloro-2-isocyanatobenzene (300 mg,1.95 mmol) and 2-cyanoethylthioamide (2-cyanoethanethioamide) (227 mg,1.95 mmol) were added to DMF (5 mL) followed by KOH (219 mg,3.90 mmol). The reaction was carried out at room temperature for 5 hours. EtOAc (20 mL) and H ₂ O (20 mL) were added to dilute, pH adjusted to 5 with aqueous hydrochloric acid (1N), washed sequentially with H ₂ O (20 ml×2) and saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was chromatographed by column chromatography (DCM/MeOH (v/v) =20/1) to give 5- ((2-chlorophenyl) amino) -3-mercaptoisothiazole-4-carbonitrile (5) (50 mg, 10%).

MS(ESI,pos.ion)m/z:268.1[M+1]⁺。

¹HNMR(DMSO-d₆)δ:8.82(s,2H),7.50(d,J＝7.8Hz,1H),7.28-7.36(m,1H),7.10-7.18(m,1H),7.04(d,J＝7.8Hz,1H).

Example 2: synthesis of 5- ((2-chlorophenyl) amino) -2-methyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (30)

Step 1: synthesis of 2-cyano-N-methylethylthioamide

2-Cyano-N-methylacetamide (2.0 g,20.39 mmol) was dissolved in toluene (20 mL), L-Lawson's reagent (4.10 g,10.19 mmol) was added at 25℃and the reaction stirred at 110℃for 0.5h. After completion of the reaction, the reaction mixture was directly dried by spin-drying, and purified by silica gel column chromatography (SiO ₂, PE: EA (v/v) =2:1) to give compound 30-1 (1.21 g, 52%).

MS(ESI,pos.ion)m/z:115.1[M+1]⁺。

Step 2: synthesis of 5- ((2-chlorophenyl) amino) -2-methyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (30)

2-Cyano-N-methylacetamide (300 mg,2.63 mmol) and 1-chloro-2-isothiocyanatobenzene (4476 mg,2.63 mmol) were dissolved in DMF (5 mL), KOH (221 mg,3.94 mmol) was added in portions and stirred overnight at room temperature. The reaction solution was diluted with water, ph=7 was adjusted with HCl (1 moL/L), extracted with ethyl acetate (20 ml×3), washed with water (20 ml×2) and saturated brine (20 ml×2), the organic phases were combined and dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, PE/EA (v/v) =1/0 to 0/1) and preparative HPLC to give 5- ((2-chlorophenyl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (30) (113 mg, 15%).

MS(ESI,pos.ion)m/z:282.0[M+1]⁺。

¹HNMR(400MHz,DMSO-d₆)δ9.23(s,1H),7.50(dd,J＝7.9,1.3Hz,1H),7.32(td,J＝7.6,1.4Hz,1H),7.13(td,J＝7.7,1.6Hz,1H),7.04(dd,J＝7.9,1.6Hz,1H),3.01(s,3H).

Example 3: synthesis of 2-ethyl-5- ((2-fluorophenyl) amino) -3-thio-2, 3-diisothiazole-4-carbonitrile (31A)

Step 1: synthesis of 2-cyano-N-ethylthioamide

2-Cyano-N-ethylacetamide (300 mg,2.68 mmol) and L.sub.Lawson reagent (541 mg,1.19 mmol) were added to toluene (5 mL). The reaction was carried out at 110℃for 1 hour. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with H ₂ O (20 ml×2) and saturated NaCl solution (20 mL) in this order, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was chromatographed by column chromatography (PET/EtOAc (v/v) =4/1) to give compound 31-1 (330 mg, 96%).

Step 2: synthesis of 2-ethyl-5- ((2-fluorophenyl) amino) -3-thio-2, 3-diisothiazole-4-carbonitrile (31A)

1-Fluoro-2-isothiocyanatobenzene (119 mg,0.78 mmol) and 2-cyano-N-ethylthioamide (100 mg,0.78 mmol) were added to DMF (5 mL), KOH (66 mg,1.17 mmol) was added thereto at room temperature, and stirred overnight. H ₂ O (20 mL) was added to dilute, extracted with EA (20 mL. Times.3), and washed with saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product purified by HPLC to give 2-ethyl-5- ((2-fluorophenyl) amino) -3-thio-2, 3-diisothiazole-4-carbonitrile (31A) (10 mg, 5%).

MS(ESI,pos.ion)m/z:279.9[M+1]⁺。

¹H NMR(DMSO-d₆)δ:9.26(br s,1H),7.21-7.32(m,1H),7.11-7.21(m,2H),7.00-7.09(m,1H),3.40-3.50(m,2H),1.23(t,J＝7.2Hz,3H).

Example 4: synthesis of 5- ((2-chlorophenyl) amino) -2-isopropyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (32)

Step 1: synthesis of 2-cyano-N-isopropyl acetamide

Ethyl 2-cyanoacetate (1.06 g,9.37 mmol) and isopropylamine (554 mg,9.37 mmol) were added to EtOH (10 mL) and reacted overnight at 70 ℃. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove excess ethanol, diluted with EtOAc (50 mL), washed with H ₂ O (20 ml×2) and saturated NaCl solution (20 mL) in this order, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was separated by column chromatography (PE/EA (v/v) =3/1) to give compound 32-1 (800 mg, 68%).

Step 2: synthesis of 2-cyano-N-isopropyl ethyl sulfonamide

2-Cyano-N-isopropyl acetamide (300 mg,2.38 mmol) and L-Lawson reagent (481 mg,1.19 mmol) were added to toluene (10 mL). The reaction was carried out at 110℃for 1 hour. The reaction was cooled to room temperature, diluted with EtOAc (20 mL), washed with H ₂ O (20 ml×2) and saturated NaCl solution (20 mL) in this order, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was chromatographed by column chromatography (PET/EtOAc (v/v) =4/1) to give compound 32-2 (300 mg, 89%).

Step 3: synthesis of 5- ((2-chlorophenyl) amino) -2-isopropyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (32)

2-Cyano-N-isopropylethanesulfonamide (300 mg,2.11 mmol) and 1-chloro-2-isothiocyanatobenzene (0.28 mL,2.11 mmol) were added to DMF (5 mL) followed by slow addition of KOH (178 mg,3.16 mmol). Stir at room temperature overnight. After the reaction, water (10 mL) was added to dilute the mixture, and the pH was adjusted to 6 with 1N hydrochloric acid. Extraction with EtOAc (30 mL), washing with saturated NaCl solution (15 mL), drying over anhydrous Na ₂SO₄, concentrating under reduced pressure, and purification of the crude product by HPLC afforded 5- ((2-chlorophenyl) amino) -2-isopropyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (32) (7.4 mg, 1%).

MS(ESI,pos.ion)m/z:309.9[M+1]⁺。

¹HNMR(DMSO-d₆)δ:9.18(br d,J＝8.2Hz,1H),7.50(d,J＝7.9Hz,1H),7.32(t,J＝7.6Hz,1H),7.13(t,J＝7.7Hz,1H),7.03(d,J＝7.7Hz,1H),3.83(br s,1H),1.27(d,J＝6.2Hz,6H).

Example 5: synthesis of 5- ((2-chlorophenyl) amino) -2-cyclopropyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (33)

Step 1: synthesis of 2-cyano-N-cyclopropylacetamide

Ethyl 2-cyanoacetate (1.06 g,9.37 mmol) and cyclopropylamine (535 mg,9.37 mmol) were added to EtOH (10 mL) and reacted overnight at 70 ℃. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove excess ethanol, diluted with EtOAc (50 mL), washed with H ₂ O (20 ml×2) and saturated NaCl solution (20 mL) in this order, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was separated by column chromatography (PE/EA (v/v) =3/1) to give compound 33-1 (82mg, 70%).

Step 2: synthesis of 2-cyano-N-cyclopropylethylthioamide

2-Cyano-N-cyclopropylacetamide (200 mg,1.61 mmol) and L-Lawson reagent (326 mg,0.81 mmol) were added to toluene (10 mL). The reaction was heated at 110℃for 1 hour. The reaction solution was cooled to room temperature, diluted with EtOAc (20 mL), washed with H ₂ O (20 ml×2) and saturated NaCl solution (20 mL) in this order, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was chromatographed by column chromatography (PE/EA (v/v) =4/1) to give compound 33-2 (180 mg, 80%).

Step 3: synthesis of 5- ((2-chlorophenyl) amino) -2-cyclopropyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (33)

2-Cyano-N-cyclopropylethylthioamide (140 mg,1.00 mmol) and 1-chloro-2-isothiocyanatobenzene (0.13 mL,1.00 mmol) were added to DMF (5 mL) followed by slow addition of KOH (84 mg,1.50 mmol). Stirred at room temperature overnight, diluted with water (10 mL) and adjusted to pH 6 with 1N hydrochloric acid. Extracted with EtOAc (30 mL), washed with saturated NaCl solution (15 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product purified by HPLC to give 5- ((2-chlorophenyl) amino) -2-cyclopropyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (33) (17 mg, 6%).

MS(ESI,pos.ion)m/z:307.8[M+1]⁺。

¹HNMR(DMSO-d₆)δ:9.56(br s,1H),7.50(dd,J＝8.0,1.2Hz,1H),7.31(td,J＝7.6,1.3Hz,1H),7.12(td,J＝7.7,1.5Hz,1H),7.02(dd,J＝7.9,1.3Hz,1H),2.79(br s,1H),0.78-0.86(m,4H).

Example 6: synthesis of 5- ((2-chlorophenyl) amino) -2-cyclobutyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (34)

Step 1: synthesis of 2-cyano-N-cyclobutylacetamide

Ethyl 2-cyanoacetate (2.00 g,17.68 mmol) and cyclobutylamine (1.89 g,26.52 mmol) were added to EtOH (15 mL) and reacted at 70℃for 12h. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove excess ethanol, and the residue was slurried with a mixed solvent of ethyl acetate and petroleum ether (1:10, 10 mL), filtered and washed with petroleum ether (10 mL) to give compound 34-1 (2.00 g, yield: 81%).

Step 2: synthesis of 2-cyano-N-cyclobutylethylthioamide

2-Cyano-N-cyclobutylacetamide (500 mg,3.62 mmol), L.Lawsonia reagent (730 mg,1.81 mmol) was added to toluene (5 mL). The reaction was carried out at 110℃for 0.5 hour. The reaction was cooled to room temperature, diluted with EtOAc (20 mL), washed with H ₂ O (20 ml×2) and saturated NaCl solution (20 mL) in this order, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was chromatographed by column chromatography (PE/EA (v/v) =4/1) to give compound 34-2 (360 mg, 65%).

Step 3: synthesis of 5- ((2-chlorophenyl) amino) -2-cyclobutyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (34)

2-Cyano-N-cyclobutylethylthioamide (200 mg,1.30 mmol) and 1-chloro-2-isothiocyanatobenzene (220 mg,1.30 mmol) were added to DMF (5 mL) followed by slow addition of KOH (110 mg,1.95 mmol). Stirred at room temperature overnight, diluted with water (10 mL) and adjusted to pH 6 with 1N hydrochloric acid. Extracted with EtOAc (30 mL), washed with saturated NaCl solution (15 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product purified by HPLC to give 5- ((2-chlorophenyl) amino) -2-cyclobutyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (34) (8.6 mg, 2%).

L MS(ESI,pos.ion)m/z:321.8[M+H]⁺.

¹HNMR(400MHz,DMSO-d6)δ9.54(d,J＝7.0Hz,1H),7.51(dd,J＝8.0,1.4Hz,1H),7.32(td,J＝7.6,1.4Hz,1H),7.14(td,J＝7.7,1.6Hz,1H),7.03(dd,J＝7.9,1.6Hz,1H),4.14(d,J＝9.5Hz,1H),2.38–2.14(m,4H),1.80–1.62(m,2H).

Example 7: synthesis of 5- ((2-chlorophenyl) amino) -2-cyclopentyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (35)

Step 1: synthesis of 2-cyano-N-cyclopentylacetamide

Ethyl 2-cyanoacetate (2.00 g,17.68 mmol) and cyclopentylamine (2.26 g,26.52 mmol) were added to EtOH (15 mL) and reacted overnight at 70 ℃. The reaction solution was cooled to room temperature, concentrated under reduced pressure to remove excess ethanol, and the residue was slurried with a mixed solvent of ethyl acetate and petroleum ether (1:10, 10 mL), filtered and washed with petroleum ether (10 mL) to give compound 35-1 (2.6 g, yield: 96%).

Step 2: synthesis of 2-cyano-N-cyclopentylethylthioamide

2-Cyano-N-cyclopentylacetamide (500 mg,3.62 mmol) and L-Lawson reagent (660 mg,1.64 mmol) were added to toluene (5 mL). The reaction was carried out at 110℃for 0.5 hour. The reaction was cooled to room temperature, diluted with EtOAc (20 mL), washed with H ₂ O (20 ml×2) and saturated NaCl solution (20 mL) in this order, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was chromatographed by column chromatography (PE/EA (v/v) =4/1) to give compound 35-2 (450 mg, 82%).

Step 3: synthesis of 5- ((2-chlorophenyl) amino) -2-cyclopentyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (35)

2-Cyano-N-cyclopentylethylthioamide (200 mg,1.19 mmol) and 1-chloro-2-isothiocyanatobenzene (200 mg,1.19 mmol) were added to DMF (5 mL) followed by slow addition of KOH (100 mg,1.78 mmol). Stirred at room temperature overnight, diluted with water (10 mL) and adjusted to pH 6 with 1N hydrochloric acid. Extracted with EtOAc (30 mL), washed with saturated NaCl solution (15 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product purified by HPLC to give 5- ((2-chlorophenyl) amino) -2-cyclopentyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (35) (4.7 mg, 1%).

L MS(ESI,pos.ion)m/z:335.8[M+H]⁺.

¹HNMR(400MHz,DMSO-d6)δ9.27(d,J＝7.7Hz,1H),7.51(dd,J＝8.0,1.4Hz,1H),7.32(td,J＝7.6,1.4Hz,1H),7.14(td,J＝7.7,1.6Hz,1H),7.04(dd,J＝7.8,1.6Hz,1H),4.04(s,1H),2.04–1.93(m,2H),1.70(dtt,J＝13.0,6.3,3.6Hz,4H),1.56(dp,J＝10.8,3.9,3.0Hz,2H).

Example 8: synthesis of 5- ((2-chlorophenyl) amino) -2-cyclohexyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (36)

Step 1: synthesis of 2-cyano-N-cyclohexylacetamide

Ethyl 2-cyanoacetate (1.00 g,8.84 mmol) and cyclohexylamine (1.75 g,17.68 mmol) were added to EtOH (20 mL). The reaction was carried out at 70℃for 25h. Concentrated under reduced pressure, EA (10 mL) was added, PE was added dropwise until a large amount of solid precipitated, and the solid was filtered to give a yellow solid (1.20 g, 81%).

MS(ESI,pos.ion)m/z:167.1[M+1]⁺。

Step 2: synthesis of 2-cyano-N-cyclohexylethylthioamide

2-Cyano-N-cyclohexylacetamide (740 mg,4.45 mmol) and L-Lawson's reagent (900 mg,2.23 mmol) were added to toluene (20 mL) and reacted at 110℃for 0.5h. Concentrated under reduced pressure, and the crude product was chromatographed by column chromatography (PE/EtOAc (v/v) =3/1) to give a pale yellow solid (600 mg, 74%).

MS(ESI,pos.ion)m/z:183.1[M+1]⁺。

Step 3: synthesis of 5- ((2-chlorophenyl) amino) -2-cyclohexyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (36)

2-Cyano-N-cyclohexylethylthioamide (300 mg,1.65 mmol) and 1-chloro-2-isothiocyanatobenzene (279 mg,1.65 mmol) were added to DMF (10 mL), KOH (139 mg,2.47 mmol) was added and reacted at room temperature for 12h. EtOAc (20 mL) was added for dilution, washed sequentially with H ₂ O (20 ml×2) and saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was chromatographed (DCM/MeOH (v/v) =20/1) by column chromatography to give 5- ((2-chlorophenyl) amino) -2-cyclohexyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (36) (18 mg, 3%).

MS(ESI,pos.ion)m/z:350.1[M+1]⁺。

¹H NMR(400MHz,DMSO-d₆)δ9.19(d,J＝8.0Hz,1H),7.51(dd,J＝8.0,1.4Hz,1H),7.32(td,J＝7.6,1.4Hz,1H),7.14(td,J＝7.6,1.6Hz,1H),7.04(dd,J＝8.0,1.6Hz,1H),1.93(d,J＝12.1Hz,2H),1.75(d,J＝12.8Hz,2H),1.60(d,J＝12.8Hz,1H),1.44(q,J＝11.8Hz,2H),1.37–1.20(m,3H),1.12(t,J＝12.4Hz,1H)

Example 9: synthesis of 5- ((2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile

1-Fluoro-2-isothiocyanatobenzene (125 mg,0.82 mmol) and 3-amino-3-thiopropionitrile (82 mg,0.82 mmol) were added to DMF (5 mL), KOH (69 mg,1.23 mmol) was added portionwise at room temperature, and stirred overnight at room temperature. 20mL of water was added to the reaction, pH=7, the aqueous phase was adjusted with HCl (1N), extracted with ethyl acetate (20 mL. Times.3), the organic phase was washed with saturated brine, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by preparative TLC (DCM/MeOH (v/v) =20/1) and preparative HPLC and lyophilized to give 5- ((2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (37) (10 mg, 5%).

MS(ESI,pos.ion)m/z:251.9[M+1]⁺。

¹H NMR(400MHz,DMSO-d₆)δ8.81(s,2H),7.25(ddd,J＝10.9,7.5,2.4Hz,1H),7.20–7.09(m,2H),7.04(ddd,J＝9.4,6.8,2.5Hz,1H).

Example 10: synthesis of 3-thio-5- ((2- (trifluoromethyl) phenyl) amino) -2, 3-dihydroisothiazole-4-carbonitrile (38-A)

1-Isothiocyanato-2- (trifluoromethyl) benzene (100 mg,0.49 mmol) and 2-cyanoethylthioamide (49 mg,0.49 mmol) were added to DMF (5 mL), KOH (41 mg,0.74 mmol) was added in portions at room temperature, and stirred overnight at room temperature. 20mL of water was added to the reaction solution, pH=7 of the aqueous phase was adjusted with HCl (1N), extracted with ethyl acetate (20 mL. Times.3), the organic phase was washed with saturated brine, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by preparative TLC (DCM/MeOH (v/v) =20/1) and preparative HPLC and lyophilized to give 3-thio-5- ((2- (trifluoromethyl) phenyl) amino) -2, 3-dihydroisothiazole-4-carbonitrile (38) (7 mg, 5%).

MS(ESI,pos.ion)m/z:301.8[M+1]⁺。

¹H NMR(400MHz,DMSO-d₆)δ8.82(s,2H),7.71(dd,J＝7.9,1.5Hz,1H),7.62(td,J＝7.7,1.5Hz,1H),7.28(t,J＝7.6Hz,1H),7.15(d,J＝7.9Hz,1H).

Example 11: synthesis of 5- ((2-chloronaphthalen-1-yl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (39-A)

Step 1: synthesis of 2-chloronaphthalene-1-amine

Naphthalene-1-amine (1000 mg,6.98 mmol) was dissolved in THF (15 mL), 1-chloropyrrolidine-2, 5-dione (930 mg,6.98 mmol) was added in portions at 0deg.C and reacted for 1h at 0deg.C. The reaction mixture was quenched with water and extracted with ethyl acetate (50 mL. Times.3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by preparative TLC (PE: EA (v/v) =5/1) to give compound 39-1 (525 mg, 42%).

MS(ESI,pos.ion)m/z:177.9[M+1]⁺。

Step 2: synthesis of 2-chloro-1-isothiocyanato naphthalene

2-Chloronaphthalen-1-amine (525 mg,2.96 mmol) was dissolved in DCM (5 mL), and a solution of thiophosgene (408 mg,3.55 mmol) in DCM (5 mL) was added dropwise under nitrogen at 0℃and reacted at the same temperature for 0.5h. The reaction was adjusted to pH with HCl (1N, 20 mL) and extracted with DCM (10 mL. Times.3). The organic phase was washed with saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product (497 mg, 76.5%) was used directly in the next reaction.

Step 3: synthesis of 5- ((2-chloronaphthalen-1-yl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (39)

2-Chloro-1-isothiocyanatonaphthalene (497 mg,2.26 mmol) and 3-amino-3-thiopropionitrile (227 mg,2.26 mmol) were dissolved in DMF (5 mL), KOH (126 mg,2.26 mmol) was added in portions, and stirred at room temperature overnight. The reaction solution was diluted with water, ph=7 was adjusted with HCl (1N), and extracted with ethyl acetate (20 ml×3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure and the crude product purified by silica gel column chromatography (SiO ₂, DCM/meoh=1/0 to 40/1) and preparative HPLC to give 5- ((2-chloronaphthalen-1-yl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (39) (117 mg, 16%).

MS(ESI,pos.ion)m/z:317.8[M+1]⁺。

¹H NMR(400MHz,DMSO-d₆)δ7.06(ddt,J＝14.8,8.4,4.1Hz,2H),6.86(d,J＝8.8Hz,1H),6.71(ddt,J＝13.2,9.0,3.9Hz,3H).

Example 12: synthesis of 5- (phenylamino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (54)

Isothiocyanate (300 mg,2.22 mmol) and 2-cyanoethylthioamide (222 mg,2.22 mmol) were dissolved in DMF (3 mL), KOH (187 mg,3.33 mmol) was added in portions and stirred overnight at room temperature. The reaction solution was diluted with water, pH=7 was adjusted with HCl (1 moL/L), extracted with ethyl acetate (20 mL. Times.3), washed with water (20 mL. Times.2) and saturated brine (20 mL. Times.2), respectively, the combined organic phases were dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, PE/EA (v/v) =1/0 to 0/1) and preparative HPLC (column: boston Prime C18150. Times. 30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% B over 8 min) to give 5- (phenylamino) -3-thio-2, 3-dihydroisothiazol-4-carbonitrile (54) (17 mg, 3%).

MS(ESI,pos.ion)m/z:234.0[M+1]⁺。

¹HNMR(400MHz,DMSO-d₆)δ8.75(s,2H),7.36(t,J＝7.9Hz,2H),7.12(t,J＝7.4Hz,1H),6.96(d,J＝7.1Hz,2H).

Example 13: synthesis of 5- ((2-methoxyphenyl) amino) -3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (55)

1-Isothiocyanato-2-methoxybenzene (200 mg,1.21 mmol) and 2-cyanoethylthioamide (121 mg,1.21 mmol) were dissolved in DMF (5 mL), KOH (102 mg,1.8 mmol) was added in portions, and stirred at room temperature for 5h. The reaction solution was diluted with water, pH=7 was adjusted with HCl (1 moL/L), extracted with ethyl acetate (20 mL. Times.3), washed with water (20 mL. Times.2) and saturated brine (20 mL. Times.2), respectively, the combined organic phases were dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, PE/EA (v/v) =1/0 to 0/1) and preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% B over 8 min) to give 5- ((2-methoxyphenyl) amino) -3-thio-2, 3-dihydroisothiazol-4-carbonitrile (55) (10 mg, 3%).

MS(ESI,pos.ion)m/z:264.0[M+1]⁺。

¹H NMR(400MHz,DMSO-d6)δ9.86(s,1H),7.47–7.38(m,1H),7.31(d,J＝7.7Hz,1H),7.23(dd,J＝8.4,1.2Hz,1H),7.06(td,J＝7.6,1.2Hz,1H),3.85(s,3H).

Example 14: synthesis of 5- ((2, 6-difluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (56)

1, 3-Difluoro-2-isothiocyanatobenzene (200 mg,1.17 mmol) and 2-cyanoethylthioamide (222 mg,1.17 mmol) were dissolved in DMF (3 mL), KOH (98 mg,1.75 mmol) was added in portions and stirred at room temperature overnight. The reaction solution was diluted with water, pH=7 was adjusted with HCl (1 moL/L), extracted with ethyl acetate (20 mL. Times.3), washed with water (20 mL. Times.2) and saturated brine (20 mL. Times.2), respectively, the combined organic phases were dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, PE/EA (v/v) =1/0 to 0/1) and preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% B over 8 min) to give 5- ((2, 6-difluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (56) (11 mg, 3.5%).

MS(ESI,pos.ion)m/z:270.0[M+1]⁺。

¹HNMR(500MHz,DMSO-d₆)δ8.92(s,2H),7.15(m,J＝8.8Hz,3H).

Example 15: synthesis of 5- (cyclohexylamino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (57)

3-Amino-3-thiopropionitrile (0.16 mL,2.00 mmol) and isothiocyanate cyclohexane (0.27 mL,2.00 mmol) were added to DMF (3 mL). The reaction was carried out at 25℃for 5 hours. Extraction with ethyl acetate (20 mL. Times.3), washing with water (20 mL. Times.2) and saturated brine (20 mL. Times.2) respectively, drying the combined organic phases with anhydrous Na ₂SO₄, concentrating under reduced pressure, slurrying the crude product with dichloromethane, filtering to give a brown solid, purifying the solid by HPLC to give 5- (cyclohexylamino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (57) (20 mg, 5%).

MS(ESI,pos.ion)m/z:240[M-1]⁺。

¹HNMR(400MHz,DMSO-d6)δ8.49(s,1H),4.87–4.77(m,1H),1.94(d,J＝10.6Hz,2H),1.79(d,J＝11.8Hz,2H),1.62(d,J＝12.6Hz,1H),1.44–1.13(m,6H).

Example 16: synthesis of N- (4-cyano-3-thio-2, 3-dihydroisothiazol-5-yl) -N-phenylacetamide (59)

Step 1: synthesis of 5- (phenylamino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile

Isothiocyanate (1.0 g,9.99 mmol) and 2-cyanoethylthioamide (1.35 g,9.99 mmol) were dissolved in DMF (10 mL), KOH (0.84 g,14.98 mmol) was added in portions and stirred overnight at room temperature. The reaction solution was diluted with water, ph=7 was adjusted with HCl (1 moL/L), extracted with ethyl acetate (50 ml×3), the organic phase was washed with water (50 ml×2) and saturated brine (50 ml×2), respectively, the combined organic phases were dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, PE/EA (v/v) =1/0 to 0/1) to give 5- (phenylamino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (322 mg, 14%).

MS(ESI,pos.ion)m/z:234.0[M+1]⁺。

Step 2: synthesis of N- (4-cyano-3-thio-2, 3-dihydroisothiazol-5-yl) -N-phenylacetamide (59)

5- (Phenylamino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (40 mg,0.17 mmol) and sodium bicarbonate (126 mg,1.49 mmol) were dissolved in DCM (3 mL), acetic anhydride (18 mg,0.23 mmol) was slowly added dropwise at 0deg.C, and then the reaction was stirred at room temperature for 1h. After dilution of the reaction with water, the organic phase was washed once with DCM (20 mL. Times.3), the organic phase was combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, PE/EA (v/v) =1/0 to 1/1) and preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% B over 8 min) to give N- (4-cyano-3-thio-2, 3-dihydroisothiazol-5-yl) -N-phenylacetamide (59) (10 mg, 21%).

MS(ESI,pos.ion)m/z:276.0[M+1]⁺。

H NMR(500MHz,DMSO-d6)δ12.03(br,1H),7.65–7.00(m,5H),2.28(s,3H).

Example 17: synthesis of N- (2-acetyl-4-cyano-3-thio-2, 3-dihydroisothiazol-5-yl) -N-phenylacetamide (59 c)

5- (Phenylamino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (40 mg,0.17 mmol) and sodium bicarbonate (126 mg,1.49 mmol) were dissolved in DCM (3 mL), acetic anhydride (18 mg,0.23 mmol) was slowly added dropwise at 0deg.C and the reaction stirred at room temperature for 1h. After the reaction mixture was diluted with water, it was extracted with DCM (20 mL. Times.3), and the organic phase was washed once with water (20 mL) and saturated brine (20 mL). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, PE/EA (v/v) =1/0 to 3/1) and preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% b over 8 min) to give N- (2-acetyl-4-cyano-3-thio-2, 3-dihydroisothiazol-5-yl) -N-phenylacetamide (59C) (6 mg, 11%).

MS(ESI,pos.ion)m/z:318.0[M+1]⁺。

¹HNMR(400MHz,DMSO-d₆)δ7.72(d,J＝6.9Hz,2H),7.60(dt,J＝14.4,7.0Hz,3H),2.25(s,3H),2.03(s,3H).

Example 18: synthesis of 5- ((2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carboxamide (60)

Step 1: synthesis of 5- ((2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile

1-Fluoro-2-isothiocyanatobenzene (300 mg,1.96 mmol) and 3-amino-3-thiopropionitrile (196 mg,1.96 mmol) were added to DMF (3 mL), KOH (165 mg,2.94 mmol) was added in portions at room temperature, and stirred overnight at room temperature. 20mL of water was added to the reaction solution, pH=7 of the aqueous phase was adjusted with HCl (1N), extracted with ethyl acetate (20 mL. Times.3), the organic phase was washed with saturated brine, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by preparative TLC (DCM/MeOH (v/v) =20/1) to give compound 60-1 (235 mg, 39%).

MS(ESI,pos.ion)m/z:251.9[M+1]⁺。

Step 2: synthesis of 5- ((2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carboxamide (60)

5- ((2-Fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (100 mg,0.40 mmol) was added to H ₂SO₄ (1 mL). The reaction was carried out at 50℃for 30min. After the reaction solution was cooled to room temperature, the reaction solution was added dropwise to the cooled saturated sodium bicarbonate solution. After quenching the reaction, the aqueous phase was extracted with DCM (10 mL. Times.2). The organic phase was washed with saturated brine and dried over anhydrous Na ₂SO₄. The crude product was purified by preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% B over 8 min) to give 5- ((2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carboxamide (60) (37 mg, 34%).

MS(ESI,pos.ion)m/z:271[M+1]⁺。

¹H NMR(500MHz,DMSO-d₆)δ9.99(s,1H),9.05(d,J＝4.0Hz,2H),7.40(d,J＝3.9Hz,1H),7.27(ddd,J＝10.6,7.6,2.0Hz,1H),7.20–7.09(m,3H).

Example 19: synthesis of 5- ((6-chloroquinolin-5-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (63)

Step 1: synthesis of 6-chloro-5-isothiocyanato quinoline

5-Amino-6-chloroquinoline (200 mg,1.12 mmol) was dissolved in DCM (3 mL), and a solution of DIEA and thiophosgene (457 mg,3.94mmol,0.3 mL) in DCM (1 mL) was added dropwise at 0deg.C and reacted at the same temperature for 0.5h. The reaction was adjusted to pH with HCl (1N, 10 mL) and extracted with DCM (10 mL. Times.3). The organic phase was washed with saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product (200 mg, 81%) was used directly in the next reaction.

Step 2: synthesis of 5- ((6-chloroquinolin-5-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (63)

6-Chloro-5-isothiocyanato-quinoline (200 mg,0.91 mmol) and 2-cyano-N-methylthioacetamide (124 mg,1.09 mmol) were dissolved in DMF (4 mL), KOH (76 mg,1.35 mmol) was added in portions and stirred overnight at room temperature. The reaction solution was diluted with water, ph=7 was adjusted with HCl (1N), and extracted with ethyl acetate (20 ml×3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure and the crude product was purified by column chromatography on silica gel (SiO 2, PE/ea=1/0 to 90/1) and preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% b over 8 min) to give 5- ((6-chloroquinolin-5-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (63) (43 mg, 14%).

MS(ESI,pos.ion)m/z:333[M+1]⁺。

¹H NMR(DMSO-d₆)δ:9.37(br d,J＝3.8Hz,1H),8.85-8.97(m,1H),8.17(br d,J＝8.3Hz,1H),7.72-7.88(m,2H),7.56(dd,J＝8.4,4.3Hz,1H),3.03(br d,J＝3.4Hz,3H)

Example 20: synthesis of 5- ((6-fluoroquinolin-5-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (64)

Step 1: synthesis of 6-fluoro-5-nitroquinoline

H ₂SO₄ (4 mL) was added dropwise to HNO ₃ (0.67 mL,10.19 mmol) at 0deg.C, followed by dropwise addition of the mixed acid to 6-fluoroquinoline (0.83 mL,6.80 mmol) at 0deg.C and stirring at room temperature overnight. TLC monitored the end of the reaction, the reaction was slowly poured into crushed ice water, the pH was adjusted to 6 with saturated sodium bicarbonate solution and extracted with ethyl acetate (20 ml×3). The organic phase was dried over anhydrous Na ₂SO₄ and concentrated under reduced pressure to give compound 64-1 (1080 mg, 83%).

Step 2: synthesis of 6-fluoro-5-aminoquinoline

6-Fluoro-5-nitroquinoline (881 mg,4.58 mmol) was dissolved in ethanol (30 mL) and water (10 mL), iron powder (1.53 g,27.51 mmol) and NH ₄ Cl (1.47 g,27.48 mmol) were added with stirring and the reaction was stirred overnight at 75 ℃. TLC monitored the end of the reaction, the reaction cooled to room temperature, filtered through celite, the filtrate concentrated under reduced pressure and extracted with ethyl acetate (20 mL x 3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO 2, PE/ea=1/0 to 3/1) to give compound 64-2 (327 mg, 44%).

MS(ESI,pos.ion)m/z:163.1[M+1]⁺。

Step 3: synthesis of 6-fluoro-5-isothiocyanato quinoline

6-Fluoro-5-aminoquinoline (197mg, 1.21 mmol) was added to water (2 mL), followed by concentrated HCl (1 mL,1.21 mmol) and thiophosgene (168 mg,0.22mmol,0.02 mL). The reaction was stirred at room temperature for 2h, TLC monitored for end of reaction, extracted with ethyl acetate (10 mL. Times.3), and the organic phase was dried over anhydrous Na ₂SO₄ and concentrated under reduced pressure to give compound 64-3 (332 mg, crude).

MS(ESI,pos.ion)m/z:205.1[M+1]⁺。

Step 4: synthesis of 5- ((6-fluoroquinolin-5-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (64)

6-Fluoro-5-isothiocyanato quinoline (332 mg,1.63 mmol) and 2-cyano-N-methylthioacetamide (222.73 mg,1.95 mmol) were dissolved in DMF (5 mL), KOH (137 mg,2.44 mmol) was added in portions, and stirred at room temperature for 5 hours. The reaction solution was diluted with water, ph=6 was adjusted with HCl (1N), and extracted with ethyl acetate (20 ml×3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure and the crude product was purified by column chromatography on silica gel (SiO ₂, PE/ea=1/0 to 0/1) and preparative HPLC (column: boston Prime C18150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% b over 8 min) to give 5- ((6-fluoroquinolin-5-yl) amino) -2-methyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (64) (248 mg, 48%).

MS(ESI,pos.ion)m/z:317.0[M+1]⁺。

¹H NMR(DMSO-d₆)δ:9.36(br s,1H),8.91(dd,J＝4.0,1.5Hz,1H),8.22(dd,J＝8.4,1.0Hz,1H),7.81-7.86(m,1H),7.71-7.78(m,1H),7.55(dd,J＝8.5,4.0Hz,1H),3.32(s,2H),3.04(s,3H).

Example 21: synthesis of 5- ((2-fluorophenyl) amino) -2-methyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (65)

Step 1: synthesis of 2-cyano-N-methylthioacetamide

2-Cyano-N-methylacetamide (500 mg,5.10 mmol) was dissolved in toluene (5 mL), the reaction was stirred at 110℃for 1h, the reaction was monitored by TLC and the crude product was chromatographed by column chromatography (PE/EA=3:1) to give compound 65-1 (479 mg, 81%).

MS(ESI,pos.ion)m/z:269.1[M+1]⁺。

Step 2: synthesis of 5- ((2-fluorophenyl) amino) -2-methyl-3-thio-2, 3-dihydroisothiazole-4-carbonitrile (65)

2-Cyano-N-methylthioacetamide (97.82 mg,0.86 mmol) and 1-fluoro-2-isothiocyanatobenzene (0.10 mL,0.82 mmol) were dissolved in DMF (3 mL), KOH (68.68 mg,1.22 mmol) was added in portions and stirred at room temperature for 5h. The reaction solution was diluted with water, ph=6 was adjusted with HCl (1N), and extracted with ethyl acetate (10 ml×3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure and the crude product was purified by column chromatography on silica gel (SiO ₂, PE/ea=1/0 to 0/1) and preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% b over 8 min) to give 5- ((2-fluorophenyl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (65) (23 mg, 10%).

MS(ESI,pos.ion)m/z:266[M+1]+.

¹H NMR(DMSO-d₆)δ:9.22(br s,1H),7.22-7.29(m,1H),7.10-7.19(m,2H),7.01-7.07(m,1H),3.01(s,3H)

Example 22: synthesis of 5- ((3-fluoropyridin-4-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (67)

Step 1: synthesis of 3-fluoro-4-isothiocyanato pyridine

4-Amino-3-fluoropyridine (100 mg,0.89 mmol) was dissolved in DCM (3 mL), DIEA (348 mg,2.68mmol,0.44 mL) and thiophosgene (307 mg,2.68mmol,0.20 mL) in DCM (3 mL) were added dropwise at 0deg.C and reacted at 0deg.C for 2h. The reaction was adjusted to pH with HCl (1N) and extracted with DCM (10 mL. Times.3). The organic phase was washed with saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product (99 mg, 71%) was used directly in the next reaction.

Step 2: synthesis of 5- ((3-fluoropyridin-4-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (67)

3-Fluoro-4-isothiocyanatopyridine (99 mg,0.64 mmol) and 2-cyano-N-methylthioacetamide (87.98 mg,0.77 mmol) were dissolved in DMF (2 mL), KOH (54.05 mg,0.96 mmol) was added in portions at 0deg.C, and stirred at room temperature overnight. The reaction solution was diluted with water, ph=6 was adjusted with HCl (1N), and extracted with ethyl acetate (10 ml×3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure and the crude product was purified by column chromatography on silica gel (SiO ₂, PE/ea=1/0 to 0/1) and preparative HPLC (column: boston Prime C18150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% b over 8 min) to give 5- ((3-fluoropyridin-4-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (67) (6.5 mg, 4%).

MS(ESI,pos.ion)m/z:267[M+1]⁺。

¹H NMR(DMSO-d₆)δ:9.42(br s,1H),8.54(d,J＝2.2Hz,1H),8.33(d,J＝5.2Hz,1H),7.16(dd,J＝7.2,5.3Hz,1H),3.00-3.07(m,1H),3.04(s,2H)

Example 23: synthesis of 5- ((5-cyano-2-fluorophenyl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (68)

Step 1: synthesis of 4-fluoro-3-isothiocyanatobenzonitrile

3-Amino-4-fluorobenzonitrile (500 mg,3.67 mmol) was dissolved in DCM (3 mL), DIEA (2.13 mL,12.85 mmol) and thiophosgene (1.48 g,12.85mmol,0.98 mL) in DCM (3 mL) were added dropwise at 0deg.C and reacted at the same temperature for 0.5h. The reaction was adjusted to pH with HCl (1N) and extracted with DCM (20 mL. Times.3). The organic phase was washed with saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product (460 mg, 70%) was used directly in the next reaction.

Step 2: synthesis of 5- ((5-cyano-2-fluorophenyl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (68)

4-Fluoro-3-isothiocyanatobenzonitrile (333 mg,1.87 mmol) and 2-cyano-N-methylthioacetamide (256 mg,2.24 mmol) were dissolved in DMF (5 mL), KOH (157 mg,2.80 mmol) was added in portions and stirred overnight at room temperature. The reaction solution was diluted with water, ph=6 was adjusted with HCl (1N), and extracted with ethyl acetate (20 ml×3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure and the crude product was purified by column chromatography on silica gel (SiO ₂, PE/ea=1/0 to 0/1) and preparative HPLC (column: boston Prime C18150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% b over 8 min) to give 5- ((5-cyano-2-fluorophenyl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (68) (95 mg, 17%).

MS(ESI,pos.ion)m/z:291.1[M+1]⁺。

¹H NMR(DMSO-d₆)δ:9.25-9.49(m,1H),7.62-7.69(m,1H),7.62-7.70(m,1H),7.52(dd,J＝10.0,8.9Hz,1H),3.03(s,3H)

Example 24: synthesis of 5- ((2- (methylsulfonyl) phenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (1)

Step 1: synthesis of 1-isothiocyanato-2- (methylsulfonyl) benzene

2- (Methylsulfonyl) aniline (200 mg,1.17 mmol) was added to water (4 mL), after which concentrated HCl (1 mL) and thiophosgene (161 mg,1.40mmol,0.11 mL) were added, the reaction was stirred at room temperature for 2h, after which time TLC monitoring the reaction was complete, filtered to give compound 1-1 (200 mg, 80%).

Step 2: synthesis of 5- ((2- (methylsulfonyl) phenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (1)

1-Isothiocyanato-2- (methylsulfonyl) benzene (200 mg,0.94 mmol) and 2-cyanothioacetamide (113 mg,1.13 mmol) were dissolved in DMF (4 mL), KOH (78.93 mg,1.41 mmol) was added in portions and stirred at 0deg.C for 5 hours. The reaction solution was diluted with water, ph=6 was adjusted with HCl (1N), and extracted with ethyl acetate (20 ml×3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, DCM/meoh=1/0 to 90/1) and preparative HPLC (column: boston Prime C18150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% b over 8 min) to give 5- ((2- (methylsulfonyl) phenyl) amino) -3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (1) (18.6 mg, 6%).

MS(ESI,pos.ion)m/z:312.0[M+1]⁺。

1H NMR(DMSO-d6)δ:8.91(s,2H),7.92(dd,J＝8.0,1.5Hz,1H),7.68(td,J＝7.8,1.5Hz,1H),7.30-7.35(m,1H),7.26(dd,J＝8.0,0.9Hz,1H),3.32(s,2H)

Example 25: synthesis of 5- ((3-chloro-2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (2)

Step 1: synthesis of 1-chloro-2-fluoro-3-isothiocyanatobenzene

3-Chloro-2-fluoroaniline (1 g,6.87 mmol) was dissolved in DCM (30 mL) at 0deg.C in a 100mL three-necked flask, DIEA (3.98 mL,24.05 mmol) was then added dropwise, and thiophosgene (2.37 g,20.61 mmol) was added slowly after stirring for 5 min. The reaction was carried out at room temperature for 2 hours. After the reaction was completed, the reaction mixture was quenched with water, extracted with DCM (30 ml×2), washed with saturated brine, dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, and the crude product was separated by column chromatography (PE/EtOAc (v/v) =20/1) to give 1-chloro-2-fluoro-3-isothiocyanatobenzene (1.15 g, 89%).

MS(ESI,pos.ion)m/z:188.0[M+1]⁺。

Step 2: synthesis of 5- ((3-chloro-2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (2)

1-Chloro-2-fluoro-3-isothiocyanatobenzene (200 mg,1.07 mmol) and 2-cyanoethylsulfanomide (107 mg,1.07 mmol) were dissolved in DMF (5 mL), KOH (72 mg,1.28 mmol) was added at 0℃and stirred at 0℃for 2 hours. After the reaction was completed, the reaction mixture was quenched with water, extracted with EtOAc (20 mL. Times.2), washed with H ₂ O (20 mL. Times.2) and saturated NaCl solution (20 mL) in this order, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was separated by column chromatography (DCM/MeOH (v/v) =20/1) to give a yellow solid which was purified by preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% B over 8 min) to give 5- ((3-chloro-2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazol-4-carbonitrile (2) (5.8 mg, 1.9%).

MS(ESI,pos.ion)m/z:286.0[M+1]⁺。

¹H NMR(CD3OD_SPE)δ:9.71(s,2H),8.12(td,J＝8.0,1.0Hz,1H),8.01(td,J＝8.0,1.0Hz,1H),7.87(td,J＝8.0,1.0Hz,1H).

Example 26: synthesis of 5- ((5-chloro-2, 3-dihydrobenzofuran-4-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (4A)

Step 1: synthesis of 5-chloro-2, 3-dihydrobenzofuran-4-amine

NCS (52 mg,0.39 mmol) was added to dry dichloromethane (5 mL) in which 2, 3-dihydrobenzofuran-4-amine (50 mg,0.37 mmol) was dissolved at 0deg.C, and the reaction mixture was stirred at 0deg.C for 30min. The reaction solution was concentrated under reduced pressure, and the crude product was separated by column chromatography (PE/EtOAc (v/v) =5/1) to give compound 4-1 (40 mg, 64%).

Step 2: synthesis of 5-chloro-4-isothiocyanato-2, 3-dihydrobenzofuran

5-Chloro-2, 3-dihydrobenzofuran-4-amine (52 mg,0.29 mmol) and DIEA (133 mg,1.03 mmol) were added to DCM (8 mL), nitrogen was replaced, and thiophosgene (102 mg,0.88 mmol) was added at 0deg.C. Then stirred at 0deg.C for 2 hours, after completion of the reaction, DCM (20 mL) was added for dilution, washed with H ₂ O (20 mL. Times.2) and saturated NaCl solution (20 mL) in this order, dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product was separated by column chromatography (PE/EtOAc (v/v) =10/1) to give compound 4-2 (52 mg, 83%).

Step 3: synthesis of 5- ((5-chloro-2, 3-dihydrobenzofuran-4-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (4A)

5-Chloro-4-isothiocyanato-2, 3-dihydrobenzofuran (30 mg,0.14 mmol) and 2-cyano-N-methylethylthioamide (16 mg,0.14 mmol) were added to DMF (5 mL) and KOH (12 mg,0.21 mmol) was added at room temperature. The reaction was stirred at room temperature overnight. H ₂ O (20 mL) was added for dilution, extracted with EA (20 mL. Times.3), washed with saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product purified by HPLC to give 5- ((5-chloro-2, 3-dihydrobenzofuran-4-yl) amino) -2-methyl-3-thioxo-2, 3-dihydroisothiazole-4-carbonitrile (4A) (6 mg, 14%).

MS(ESI,pos.ion)m/z:323.9[M+1]⁺。

¹HNMR(DMSO-d₆)δ:9.24(br s,1H),7.19(d,J＝8.5Hz,1H),6.54(d,J＝8.5Hz,1H),4.56(t,J＝8.8Hz,2H),2.93-3.10(m,5H).

Example 27: synthesis of tert-butyl 5-chloro-4- ((4-cyano-3-thioxo-2, 3-dihydroisothiazol-5-yl) amino) indoline-1-carboxylate (7B)

Step 1: synthesis of 4-nitro-1H-indole-1-carboxylic acid tert-butyl ester

4-Nitro-1H-indole (2.00 g,12.34 mmol) was added to DCM (20 ml) at room temperature and DMAP (150 mg,1.24 mmol) was slowly added and stirred at room temperature for 1H. The reaction mixture was quenched with water, extracted with methylene chloride (20 ml. Times.2), dried over anhydrous sodium sulfate, concentrated and purified by column chromatography to give compound 7-1 (2.8 g, 90%).

MS(ESI,pos.ion)m/z:263.26[M+1]⁺。

Step 2: synthesis of tert-butyl 4-aminoindoline-1-carboxylate

4-Nitro-1H-indole-1-carboxylic acid tert-butyl ester (2.00 g,7.63 mmol) was added to MeOH (20 mL), then Pd/C (200 mg,1.88 mmol) was added, replaced 3 times with hydrogen, and then reacted at 60℃for 2H. The reaction solution was cooled to room temperature, and concentrated by filtration to give compound 7-2 (1.40 g, 78%).

MS(ESI,pos.ion)m/z:235.1[M+1]⁺。

Step 3: synthesis of tert-butyl 4-amino-5-chloroindoline-1-carboxylate

4-Aminoindoline-1-carboxylic acid tert-butyl ester (500 mg,2.13 mmol) was added to DCM (5 mL) and NCS (284 mg,2.13 mmol) was added at room temperature. The reaction was carried out at 25℃for 2 hours. The reaction was quenched with water, extracted with ethyl acetate (20 ml×2), the organic phase was collected, and applied to a column (PE/ea=3:1) to give compound 7-3 (344 mg, 60%).

MS(ESI,neg.ion)m/z:269.7[M-1]⁺。

Step 4: synthesis of tert-butyl 5-chloro-4-isothiocyanato indoline-1-carboxylate

Tert-butyl 4-amino-5-chloroindoline-1-carboxylate (100 mg,0.37 mmol) was added to DCM (2 ml), DIEA (95.64 mg,0.74 mmol) was added, cooled to 0deg.C, and thiophosgene (76.19 mg,0.74 mmol) was added. Stirring at 0deg.C for 2 hr, quenching with water, extracting with dichloromethane (20 mL×2), drying with anhydrous sodium sulfate, concentrating, and loading onto column (PE/EA=10:1) to obtain the final product, i.e. tert-butyl 5-chloro-4-isothiocyanato-indoline-1-carboxylate (80 mg, 69%).

MS(ESI,pos.ion)m/z:311.8[M+1]⁺。

Step 5: synthesis of tert-butyl 5-chloro-4- ((4-cyano-3-thioxo-2, 3-dihydroisothiazol-5-yl) amino) indoline-1-carboxylate (7B)

Tert-butyl 5-chloro-4-isothiocyanato-1-carboxylate (89 mg,0.29 mmol) was added to DMF (2 mL), then KOH (24.1 mg,0.42 mmol) was added, 3-amino-3-thiopropionitrile (31.94 mg,0.32 mmol) was added, stirred at 25℃for 5 hours, quenched with water, extracted with ethyl acetate (20 mL. Times.2), washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to give tert-butyl 5-chloro-4- ((4-cyano-3-thioxo-2, 3-dihydroisothiazol-5-yl) amino) indoline-1-carboxylate (30.91 mg, 27%) as the target compound purified by HPLC.

MS(ESI,pos.ion)m/z:409.1[M+1]⁺。

1H NMR(DMSO-d6)δ:8.27(s,1H),7.27-7.49(m,1H),7.25(d,J＝8.4Hz,1H),3.91(br t,J＝8.7Hz,1H),3.82-3.97(m,1H),2.85(br t,J＝8.6Hz,2H),1.50(s,9H).

Example 28: synthesis of 5- ((2-fluorophenyl) amino) -2- (2-hydroxyethyl) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (13)

Step 1 Synthesis of 2- ((tert-butyldimethylsilyl) oxy) ethan-1-amine

TBSCl (3.46 mL,20.00 mmol) was slowly added dropwise to a solution of 2-aminoethan-1-ol (1.21 mL,20 mmol) in DCM (20 mL) at 0deg.C, and after the addition, the mixture was stirred at room temperature for 1h. H ₂ O (20 mL) was added to wash, and the organic layer was washed with saturated brine (10 mL), dried over anhydrous Na ₂SO₄, filtered, and concentrated under reduced pressure to give crude (2.60 g, 74%).

Step 2 Synthesis of N- (2- ((tert-butyldimethylsilyl) oxy) ethyl) -2-cyanoacetamide

2- ((Tert-Butyldimethylsilyl) oxy) ethan-1-amine (2.60 g,14.86 mmol) and ethyl cyanoacetate (1.59 mL,14.86 mmol) were added to EtOH (20 mL) and reacted at 80℃for 12h. After the completion of the reaction, the crude product (3.60 g, 99%) was obtained by concentration under reduced pressure.

Step 3:N Synthesis of- (2- ((tert-butyldimethylsilyl) oxy) ethyl) -2-cyanoethylthioamide

N- (2- ((tert-Butyldimethylsilyl) oxy) ethyl) -2-cyanoacetamide (3.60 g,14.85 mmol) and L.sub.Lawson reagent (3.00 g,7.43 mmol) were added to toluene (20 mL) and reacted at 120℃for 3h. The reaction mixture was diluted with H ₂ O (20 mL) and extracted with ethyl acetate (20 mL. Times.2). The organic layer was washed with saturated brine (20 mL), dried over anhydrous Na ₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (PE/EA (v/v) =4/1) to give N- (2- ((tert-butyldimethylsilyl) oxy) ethyl) -2-cyanoethylthioamide (0.70 g, 18%).

MS(ESI,pos.ion)m/z:259.2[M+1]⁺。

Step 4 Synthesis of 2- (2- ((tert-butyldimethylsilyl) oxy) ethyl) -5- ((2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile

N- (2- ((tert-Butyldimethylsilyl) oxy) ethyl) -2-cyanoethylthioamide (0.7 g,2.71 mmol) and 1-fluoro-2-isothiocyanatobenzene (0.30 mL,2.46 mmol) were added to DMF (3 mL) and KOH (151 mg,2.71 mmol) was added and stirred at room temperature for 18h. The reaction mixture was diluted with H ₂ O (15 mL) and extracted with ethyl acetate (15 mL. Times.2). The organic layer was washed with saturated brine (15 mL), dried over anhydrous Na ₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (PE/EA (v/v) =10/1) to give 2- (2- ((tert-butyldimethylsilyl) oxy) ethyl) -5- ((2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (67 mg, 7%).

MS(ESI,pos.ion)m/z:410.1[M+1]⁺。

Step 5 Synthesis of 5- ((2-fluorophenyl) amino) -2- (2-hydroxyethyl) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (13) 2- (2- ((tert-butyldimethylsilyl) oxy) ethyl) -5- ((2-fluorophenyl) amino) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (20 mg,0.05 mmol) was added to TBAF (2 mL,6.81 mmol) and stirred at room temperature for 3h. The reaction mixture was diluted with H ₂ O (10 mL) and extracted with ethyl acetate (10 mL. Times.2). The organic layer was washed with saturated brine (10 mL), dried over anhydrous Na ₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by preparative chromatography (formic acid conditions) to give the compound 5- ((2-fluorophenyl) amino) -2- (2-hydroxyethyl) -3-thio-2, 3-dihydroisothiazole-4-carbonitrile (13) (8 mg, 55%).

MS(ESI,pos.ion)m/z:296.0[M+1]⁺。

¹H NMR(DMSO-d₆)δ:9.20(br s,1H),7.23-7.30(m,1H),7.11-7.21(m,2H),7.02-7.08(m,1H),3.59-3.61(m,2H),3.43(s,2H).

Example 29: synthesis of 5- ((2-fluorophenyl) amino) -3-thio-2- (2, 2-trifluoroethyl) -2, 3-dihydroisothiazole-4-carbonitrile (66A)

Step 1: synthesis of 2-cyano-N- (2, 2-trifluoroethyl) acetamide

2-Cyanoacetic acid (250 mg,2.94 mmol), 2-trifluoroethan-1-amine (433 mg,4.41 mmol), a 50% solution of T ₃ P in EA (2.55 g,8.00 mmol) and DIEA (1.13 g,8.82 mmol) were added to EA (5 mL) and stirred overnight at room temperature. The reaction solution was extracted with EA (20 mL. Times.3), and the organic phases were combined, washed with water (20 mL. Times.2) and saturated brine (20 mL. Times.1) in this order, dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give 2-cyano-N- (2, 2-trifluoroethyl) acetamide (519 mg, crude).

MS(ESI,pos.ion)m/z:167.0[M+1]⁺。

Step 2: synthesis of 2-cyano-N- (2, 2-trifluoroethyl) ethylthioamide

2-Cyano-N- (2, 2-trifluoroethyl) acetamide (300 mg,1.8 mmol) was dissolved in toluene (5 mL), lawson's reagent (264 mg,0.9 mmol) was added at room temperature, and the reaction was stirred at 110℃for 1 hour. After completion of the reaction, the reaction mixture was directly dried by spin-drying, and the crude product was purified by column chromatography on silica gel (PE: EA (v/v) =2:1) to give 2-cyano-N- (2, 2-trifluoroethyl) ethylthioamide (302 mg, 92%).

MS(ESI,pos.ion)m/z:183.0[M+1]⁺。

Step 3: synthesis of 5- ((2-fluorophenyl) amino) -3-thio-2- (2, 2-trifluoroethyl) -2, 3-dihydroisothiazole-4-carbonitrile (66A)

2-Cyano-N- (2, 2-trifluoroethyl) ethylthioamide (302 mg,1.66 mmol) and 1-fluoro-2-isothiocyanatobenzene (254 mg,1.66 mmol) were added to DMF (3 mL), KOH (140 mg,2.49 mmol) was added, and stirred overnight at room temperature. The reaction solution was diluted with water, ph=7 was adjusted with aqueous hydrochloric acid (1N), extracted with ethyl acetate (20 ml×3), the combined organic phases were washed successively with water (20 ml×2) and saturated brine (20 ml×2), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was chromatographed on silica gel (PE/EA (v/v) =3/1) and purified by preparative HPLC to give 5- ((2-fluorophenyl) amino) -3-thio-2- (2, 2-trifluoroethyl) -2, 3-dihydroisothiazole-4-carbonitrile (66A) (58 mg, 30%).

MS(ESI,pos.ion)m/z:334.0[M+1]⁺。

1HNMR(DMSO-d6)δ:9.69(br s,1H),7.26-7.36(m,1H),7.14-7.26(m,2H),7.04-7.13(m,1H),4.24-4.46(m,2H).

Example 30: synthesis of 5- [ (3-chloro-2-fluorophenyl) amino ] -3-thio-2- (2, 2-trifluoroethyl) -2, 3-dihydroisothiazole-4-carbonitrile (71)

Step 1: synthesis of 2-cyano-N- (2, 2-trifluoroethyl) acetamide

Cyanoacetic acid (500 mg,5.88 mmol), 2-trifluoroethylamine (0.70 mL,8.82 mmol), DIEA (2.92 mL,17.63 mmol) and 1-propylphosphoric anhydride (T3P) (4.24 mL,15.99 mmol) were dissolved in ethyl acetate (5 mL) and stirred at room temperature overnight. After completion of the TLC monitoring, 20mL of water was added to the reaction mixture, followed by extraction with ethyl acetate (20 mL. Times.3). The organic phase was washed with saturated NaCl solution (20 ml. Times.3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give crude 2-cyano-N- (2, 2-trifluoroethyl) acetamide (239 mg, 25%) which was used directly in the next reaction.

Step 2: synthesis of 2-cyano-N- (2, 2-trifluoroethyl) thioacetamide

2-Cyano-N- (2, 2-trifluoroethyl) acetamide (239 mg,1.44 mmol) and L-Lawson reagent (29 mg,0.72 mmol) were dissolved in 6ml toluene, stirred at 110℃for 1h, and after completion of TLC monitoring the reaction, the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (SiO ₂, PE/ea=1/0 to 3/1) to give 2-cyano-N- (2, 2-trifluoroethyl) acetamide (183mg, 70%).

Step 3: synthesis of 1-chloro-2-fluoro-3-isothiocyanatobenzene

3-Chloro-2-fluoroaniline (150 mg,1.03mmol,0.11 mL) was dissolved in 4mL of dichloromethane, DIEA (0.60 mL,3.61 mmol) and thiophosgene (355 mg,3.09mmol,0.24 mL) in DCM (3 mL) were added dropwise at 0deg.C and reacted at 0deg.C for 1 hour. The reaction was adjusted to pH with HCl (1N) and extracted with DCM (20 mL. Times.3). The organic phase was washed with saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give a crude product of 1-chloro-2-fluoro-3-isothiocyanatobenzene (150 mg, 76%), which was used directly in the next reaction.

Step 4: synthesis of 5- [ (3-chloro-2-fluorophenyl) amino ] -3-thio-2- (2, 2-trifluoroethyl) -2, 3-dihydroisothiazole-4-carbonitrile (71)

1-Chloro-2-fluoro-isothiocyanatobenzene (137 mg,0.73 mmol) and 2-cyano-N- (2, 2-trifluoroethyl) thioacetamide (153 mg,0.84 mmol) were dissolved in DMF (4 mL), potassium hydroxide (61 mg,1.09 mmol) was added in portions at 0℃and stirred at 0℃for 1 hour. After TLC monitoring the reaction, the reaction was diluted with water, ph=6 was adjusted with HCl (1N), and extracted with ethyl acetate (20 ml×3). The organic phase was dried over anhydrous Na ₂SO₄, concentrated under reduced pressure and the crude product was purified by column chromatography on silica gel (SiO 2, PE/ea=1/0 to 0/1) and preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% b over 8 min) to give 5- [ (3-chloro-2-fluorophenyl) amino ] -3-thio-2- (2, 2-trifluoroethyl) -2, 3-isothiazole-4-carbonitrile (71) (4.2 mg, 2%).

MS(ESI,pos.ion)m/z:368.0[M+1]⁺。

¹HNMR(DMSO-d₆):9.76(br s,1H),7.34(br s,1H),7.22(t,J＝8.0Hz,1H),7.09(br s,1H),4.32(br d,J＝6.8Hz,2H)

Example 31: synthesis of 5- ((3-chloro-2-fluorophenyl) amino) -3-thio-2- (3, 3-trifluoropropyl) -2, 3-dihydroisothiazole-4-carbonitrile (69)

Step 1: synthesis of 2-cyano-N- (3, 3-trifluoropropyl) acetamide

3, 3-Trifluoroprop-1-amine hydrochloride (1 g,6.69 mmol), cyanoacetic acid (254 mg,10.04 mmol), T ₃ P (50% by mass in EA solution, 12.8g,20.07 mmol) were dissolved in EA (20 mL), DIEA (2.59 g,20.07 mmol) was added dropwise and stirred overnight at room temperature. The reaction solution was diluted with EA (50 mL), washed with brine (100 mL), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give a crude product of 2-cyano-N- (3, 3-trifluoropropyl) acetamide (751 mg, 62%), which was used directly in the next reaction.

Step 2: synthesis of 2-cyano-N- (3, 3-trifluoropropyl) thioacetamide

2-Cyano-N- (3, 3-trifluoropropyl) acetamide (756 mg,4.2 mmol) and L-Lawson reagent (849 mg,2.1 mmol) were added to 20mL of toluene and reacted at 110℃for 1 hour. Concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, PE/ea=1/0 to 2/1) to give 2-cyano-N- (3, 3-trifluoropropyl) thioacetamide (430 mg, 52%).

Step 3: synthesis of 1-chloro-2-fluoro-3-isothiocyanatobenzene

3-Chloro-2-fluoroaniline (500 mg,3.43 mmol) was dissolved in DCM (10 mL), DIEA (470 mg,4.12 mmol) and thiophosgene (666 mg,5.15 mmol) in DCM (3 mL) were added dropwise under nitrogen at 0deg.C and reacted at 0deg.C for 30min. The reaction was diluted with DCM (20 mL), then washed with saturated NaCl solution (20 mL), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and the crude product purified by silica gel column chromatography (SiO ₂, PE/ea=1/0 to 10/1) to give 1-chloro-2-fluoro-3-isothiocyanatobenzene (452 mg, 70%).

Step 4: synthesis of (Z) -3- ((3-chloro-2-fluorophenyl) amino) -2-cyano-3-mercapto-N- (3, 3-trifluoropropyl) prop-2-enamide

1-Chloro-2-fluoro-3-isothiocyanatobenzene (411 mg,2.19 mmol) and 2-cyano-N- (3, 3-trifluoropropyl) thioacetamide (430 mg,2.19 mmol) were dissolved in DMF (10 mL), potassium hydroxide (185 mg,3.29 mmol) was added in portions and stirred at 0deg.C for 3 hours. The reaction solution was diluted with water, ph=6 was adjusted with HCl (1N), extracted with EA (20 ml×3), brine (50 mL) was washed, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product of (Z) -3- ((3-chloro-2-fluorophenyl) amino) -2-cyano-3-mercapto-N- (3, 3-trifluoropropyl) prop-2-enamide (50 mg, 6%).

MS(ESI,pos.ion)m/z:384.0[M+1]⁺。

Step 5: synthesis of 5- ((3-chloro-2-fluorophenyl) amino) -3-thio-2- (3, 3-trifluoropropyl) -2, 3-dihydroisothiazole-4-carbonitrile (69)

(Z) -3- ((3-chloro-2-fluorophenyl) amino) -2-cyano-3-mercapto-N- (3, 3-trifluoropropyl) prop-2-enamide (50 mg,0.13 mmol) was dissolved in EA (10 mL) and elemental iodine (66 mg,0.26 mmol) was added at 0deg.C. The reaction was carried out at 0℃for 2h. The reaction solution was diluted with EA (20 mL), washed with aqueous sodium thiosulfate (20 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (SiO ₂, DCM/MeOH=1/0 to 90/1) and preparative HPLC (column: boston Prime C18 150*30mm*5um;mobile phase: [ water-ACN ]; gradient:15% -45% B over 8 min) to give 5- ((3-chloro-2-fluorophenyl) amino) -3-thio-2- (3, 3-trifluoropropyl) -2, 3-dihydroisothiazole-4-carbonitrile (69) (9 mg, 55%).

MS(ESI,pos.ion)m/z:382.0[M+1]⁺。

¹HNMR(DMSO-d6)δ:9.38(br s,1H),7.27-7.40(m,1H),7.20(t,J＝8.1Hz,1H),6.97-7.11(m,1H),3.63(br t,J＝6.6Hz,2H),2.64-2.81(m,2H).

Biological examples

Biological test method and results

Preparation of SAM-TIR lysate

NRK1-HEK293T cells (HEK 293T cell line overexpressing the mouse Nrk1 gene, stably transformed cell line customized by Yinqiao, beijing) were seeded at 10X 10 ⁶ cells/plate into 150mm dishes (Corning; 430599) containing 25mL of growth medium. The following day, 15 μg of human SARM1 expression plasmid (SARM 1408-724 expression plasmid customized by the sense of Fabria in Beijing) or control vector (pCMV 3 control vector, cat: CV011, sense of Fabria in Beijing, https:// cn.sinobiotic logical. Com/cdna-clone/contol-vector-pCMV 3-uniagged-CV 011) was first premixed with 45 μ l X-TREMEGENE DNA transfection reagent (Roche product # 06365787001) and 750 μl OptiMEM (Gibco 31985062), and then the mixture was directly added to cells to transfect the cells. At the time of transfection, 250. Mu.l of 100mM nicotinamide riboside (Ron reagent; R056456-1g; CAS:23111-00-4; molecular weight: 290.7; storage at room temperature) was added to each dish to minimize toxicity from SAM-TIR overexpression. 48 hours after transfection, cells were collected by washing 3-4 times with cold PBS. Cells were resuspended in 0.5ml PBS containing protease inhibitors (Complete protease inhibitor cocktail, roche product # 11873580001). Cell lysates (Ningbo Xinzhi ultrasonic breaker, power 14%, ultrasonic for 5 min) were prepared by ultrasonic treatment. The lysate was centrifuged at 12500rpm for 10 minutes (Eppendorf centrifuge 5425R) at 4 ℃ to remove cell debris, the protein concentration was determined by the biquinine formate (BCA) method and used to normalize the lysate concentration, and an aliquot of the supernatant was stored at-80 ℃ for later use.

NRK1-HEK293T cells:

Host cell: HEK293T cells

Transfer plasmid information: N-Flag-linker-mouse NRK1

Human SARM1 expression plasmid: nt-Strep-TEV-human SARM1 (408-724)

2. Luminescence-based SARM1 SAM-TIR NAD enzyme Activity assay

The assay was a modification of the NAD+/NADH-GLOTM assay (Promega G9071, promega). In this assay, NAD+ circulating enzymes convert NAD+ to NADH. In the presence of NADH, the reductase enzymatically converts the pro-luciferin reductase substrate to luciferin. Fluorescein was detected using ULTRA-GLOTM rLuciferase and the chemiluminescent intensity was proportional to the amount of NAD+ and NADH in the sample. Under the present assay conditions, the amounts of nad+ and NADH present in the lysate are not detected with the assay, excluding any endogenous contribution to the final nad+ detected.

The assay was set up as follows: 40nl of candidate inhibitor (maximum concentration 200. Mu.M, 0.4% DMSO), 0.25. Mu.g lysate (5. Mu.l) and 5. Mu.l 400nM NAD+. The reaction was incubated at 37℃for 60 minutes, then 10. Mu.l of NAD+/NADH-GLOTM detection reagent was added. After 30 minutes at room temperature, the luminencesignal value was read using Envision Xcite. IC50 was calculated by fitting% inhibition values for complex concentrations and log-to-nonlinear regression (dose response-variable slope) with GraphPad 8.0.

(WO2019236890)

Results:

From the above results, it can be seen that the compounds of the present invention have SARM 1-inhibiting activity. Compounds 5 and 37 of the present invention exhibited significantly improved inhibitory activity (3.8-fold and 5.3-fold, respectively) relative to positive controls 1 and 2, respectively.

Various modifications of the application, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this disclosure (including all patents, patent applications, journal articles, books, and any other publications) is hereby incorporated by reference in its entirety.

Claims

A compound of formula I:

Or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof,

Wherein:

R ¹ is selected from -CN、-NO₂、-C(O)R^1a、-S(O)₂R^1a、-CONR^1aR^1b、-S(O)₂NR^1aR^1b、-C(＝NR^1a)NR^1aR^1b、-CO₂R^1a、3 to 7 membered saturated or partially unsaturated carbocyclyl, 4 to 7 membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5 or 6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and 8 to 10 membered bicyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the carbocyclyl, heterocyclyl, aryl and heteroaryl are each substituted with 0,1, 2,3 or 4R ^x;

r ² is selected from hydrogen, optionally substituted C _1-6 aliphatic, -C (O) R ^2a、-S(O)₂R^2a, and-CO ₂R^2a;

R ³ is- (CH ₂)_n Cy) and n is 0, 1 or 2;

Or alternatively

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

Cy is selected from the group consisting of 3-to 7-membered saturated or partially unsaturated carbocyclyl, 8-to 10-membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and 8-to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the carbocyclyl, heterocyclyl, aryl and heteroaryl are each substituted with 0, 1,2, 3 or 4R ^x;

R ⁴ is selected from the group consisting of hydrogen, optionally substituted C _1-6 aliphatic, -C (O) R ^4a、-S(O)₂R^4a、-CO₂R^4a, 3 to 7 membered saturated or partially unsaturated carbocyclyl, 4 to 7 membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5 or 6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the carbocyclyl, heterocyclyl, aryl and heteroaryl are each substituted with 0, 1,2,3 or 4R ^x; and

Each R ^x is independently selected from halogen, -CN, -NO ₂, -OH, -O-optionally substituted C _1-6 aliphatic, -SH, -S-optionally substituted C _1-6 aliphatic 、-NR^3aR^3b、-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a、-CONR^3aR^3b、-N(R^3a)SO₂R^3b、-N(R^3a)C(O)R^3b、 optionally substituted C _1-6 aliphatic, optionally substituted 5-to 6-membered saturated or partially unsaturated heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur, and optionally substituted 8-to 10-membered heteroaryl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur;

R ^1a、R^1b、R^2a、R^3a、R^3b and R ^4a are each independently hydrogen, optionally substituted C _1-6 aliphatic, optionally substituted phenyl, or optionally substituted 3-to 7-membered saturated or partially unsaturated carbocycle; or alternatively

R ^1a and R ^1b, and/or R ^3a and R ^3b, together with the nitrogen atom to which they are attached, form a 3-to 6-membered saturated or partially unsaturated heterocyclic ring;

provided that the compound is not
The compound of claim 1, R ¹ is selected from -CN、-NO₂、-C(O)R^1a、-S(O)₂R^1a、-CONR^1aR^1b、-S(O)₂NR^1aR^1b、-C(＝NR^1a)NR^1aR^1b、-CO₂R^1a、C_3-7 cycloalkyl, 4-to 6-membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, phenyl, naphthyl, 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the cycloalkyl, heterocyclyl, phenyl, naphthyl and heteroaryl are each substituted with 0, 1,2, 3 or 4R ^x;

Preferably, R ¹ is selected from -CN、-NO₂、-C(O)R^1a、-S(O)₂R^1a、-CONR^1aR^1b、-S(O)₂NR^1aR^1b、-C(＝NR^1a)NR^1aR^1b、-CO₂R^1a、 phenyl, a 5 or 6 membered heteroaryl group having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the phenyl and heteroaryl groups are each substituted with 0, 1,2,3 or 4R ^x.

Preferably, R ¹ is selected from: -CN, -S (O) ₂NH₂ and-S (O) ₂N(CH₃)₂、-CO₂ H,
The compound of claim 1 or 2, wherein R ² is selected from H, optionally substituted C _1-6 alkyl, -C (O) R ^2a、-S(O)₂R^2a, and-CO ₂R^2a, preferably H.
A compound according to any one of claims 1-3, wherein R ³ is-CH ₂-Cy、-(CH₂)₂ -Cy or Cy.
The compound of any one of claims 1-4, wherein Cy is selected from C _3-7 cycloalkyl, an 8-to 10-membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, C _6-10 aryl, a 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8-to 10-membered bicyclic heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the cycloalkyl, heterocyclyl, aryl, and heteroaryl are each substituted with 0, 1, 2,3, or 4R ^x.
The compound according to any one of claims 1-5, wherein R ⁴ is selected from hydrogen, optionally substituted C _1-6 aliphatic, -C (O) R ^4a、-S(O)₂R^4a、-CO₂R^4a、C_3-7 cycloalkyl, 4-to 6-membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl are each substituted with 0,1, 2,3 or 4R ^x;

Preferably, R ⁴ is C _1-6 alkyl, H、-CH₃、-CH₂CH₃、-CH₂CH₂CH₃、-CH(CH₃)₂、 substituted with halogen, or a group selected from:
a compound according to any one of claims 1 to 6 wherein R ^x is selected from the following groups:

a) Halogen, -CN, -NO ₂, -OH and-SH;

b) -O-optionally substituted C _1-6 aliphatic and-S-optionally substituted C _1-6 aliphatic;

c)-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-NR^3aR^3b、-CONR^3aR^3b、-CO₂R^3a、-N(R^3a)SO₂R^3b and-N (R ^3a)C(O)R^3b, wherein R ^3a、R^3b are each as defined in any one of claims 1 to 6;

d) Optionally substituted C _1-6 aliphatic, preferably optionally substituted C _1-6 alkyl, more preferably optionally substituted C _1-4 alkyl;

e) Optionally substituted 5-or 6-membered saturated or partially unsaturated heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur; and

F) Optionally substituted 8-to 10-membered heteroaryl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably optionally substituted 9-membered heteroaryl having 1-2 heteroatoms independently selected from nitrogen, oxygen and sulfur;

Preferably, R ^x is selected from halogen, -CN, -NO ₂, -OH, -O-optionally substituted C _1-6 alkyl, -SH, -S-optionally substituted C _1-6 alkyl 、-NR^3aR^3b、-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a、-CONR^3aR^3b、-N(R^3a)SO₂R^3b、-N(R^3a)C(O)R^3b and optionally substituted C _1-6 alkyl.
The compound of any one of claims 1-7, wherein R ³ is selected from
The compound of any one of claims 1-8, wherein R ^1a、R^1b、R^2a、R^3a、R^3b and R ^4a may each independently be hydrogen or optionally substituted C _1-6 alkyl, optionally substituted phenyl, or optionally substituted C _3-7 cycloalkyl, preferably hydrogen or optionally substituted C _1-6 alkyl, more preferably H and optionally substituted C _1-4 alkyl, even more preferably H and CH ₃.
A compound according to any one of claims 1-9, wherein:

R ¹ is selected from -CN、-NO₂、-CONR^1aR^1b、-S(O)₂NR^1aR^1b、-CO₂R^1a、 phenyl and a 5 or 6 membered heteroaryl having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the phenyl and heteroaryl are each substituted with 0, 1,2,3 or 4R ^x;

R ² is selected from hydrogen, optionally substituted C _1-6 alkyl, -C (O) R ^2a, and-S (O) ₂R^2a;

R ³ is- (CH ₂)_n Cy) and n is 0, 1 or 2, preferably 0;

Cy is selected from C _3-7 cycloalkyl, 8 to 10 membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5 or 6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, and 8 to 10 membered bicyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl are each substituted with 0, 1, 2,3 or 4R ^x;

r ⁴ is selected from hydrogen, optionally substituted C _1-6 alkyl, -C (O) R ^4a、-S(O)₂R^4a、C_3-7 cycloalkyl, 4-to 6-membered saturated or partially unsaturated monocyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl are each substituted with 0, 1,2,3 or 4R ^x; and

Each R ^x is independently selected from halogen, -CN, -NO ₂, -OH, -O-optionally substituted C _1-6 alkyl, -SH, -S-optionally substituted C _1-6 alkyl 、-NR^3aR^3b、-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a、-CONR^3aR^3b, and optionally substituted C _1-6 alkyl;

R ^1a、R^1b、R^2a、R^3a、R^3b and R ^4a are each independently hydrogen, optionally substituted C _1-6 alkyl.
The compound of any one of claims 1-10, wherein R ¹ is selected from-CN, -NO ₂、-CONR^1aR^1b、-S(O)₂NR^1aR^1b, and a 5 or 6 membered heteroaryl having 1-3 (e.g., 1,2, or 3) heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the heteroaryl is substituted with 0, 1,2, 3, or 4R ^x;

Preferably, R ¹ is selected from —cn and an unsubstituted 5 membered heteroaryl ring having 2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur;

More preferably, R ¹ is-CN, More preferably-CN orEven more preferably-CN.
The compound of any one of claims 1-11, wherein R ² is selected from hydrogen, optionally substituted C _1-6 alkyl, -C (O) R ^2a, and-S (O) ₂R^2a;

preferably, R ² is selected from hydrogen and optionally substituted C _1-4 alkyl (preferably unsubstituted C _1-4 alkyl, especially CH ₃);

More preferably, R ² is H.
The compound of any one of claims 1-12, wherein R ¹ is-CN and R ² is H.
The compound of any one of claims 1-13, wherein R ³ is Cy.
The compound of any one of claims 1-14, wherein Cy is selected from C _3-6 cycloalkyl, an 8-to 10-membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, phenyl, naphthyl, a 5-or 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8-to 10-membered bicyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the cycloalkyl, heterocyclyl, phenyl, naphthyl, and heteroaryl are each substituted with 0,1,2, 3, or 4R ^x;

preferably, cy is selected from:

Phenyl substituted with 0, 1,2, 3 or 4R ^x;

Naphthyl substituted by 0, 1,2, 3 or 4R ^x;

A 5 or 6 membered heteroaryl substituted with 0, 1,2,3 or 4R ^x groups having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur;

8 to 10 membered bicyclic heteroaryl substituted with 0, 1,2, 3 or 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably 9 or 10 membered bicyclic heteroaryl having 1,2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein preferably the bicyclic heteroaryl is a fused ring system of a benzene ring and a 5 or 6 membered heteroaryl, wherein all heteroatoms are in the 5 or 6 membered heteroaryl ring and are not common atoms; and

8-To 10-membered (especially 9-or 10-membered) saturated or partially unsaturated bicyclic heterocyclyl substituted with 0, 1,2, or 3R ^x having 1-3 (e.g., 1,2, or 3) heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein preferably the bicyclic heterocyclyl is a fused ring system of a benzene ring and a 5-or 6-membered saturated or partially unsaturated heterocyclyl, wherein the heteroatoms are all in the 5-or 6-membered saturated or partially unsaturated heterocyclyl ring and are not common atoms;

preferably, cy is selected from:

Preferably, cy is selected from
The compound of any one of claims 1-15, wherein R ⁴ is selected from the following groups:

a)H；

b1 Unsubstituted C _1-6 alkyl, more preferably C _1-3 alkyl;

b2 C _1-6 alkyl (preferably C _1-4 alkyl) substituted with a group selected from: -OH, -SH, and 1,2, 3 or more halogens;

c) -C (O) R ^4a and-S (O) ₂R^4a, wherein R ^4a is as defined in any one of claims 1 to 15, preferably optionally substituted C _1-6 alkyl, more preferably optionally substituted C _1-4 alkyl, even more preferably CH ₃; and

D) The following groups each substituted with 0, 1,2, 3 or 4R ^x:

d1 C _3-7 cycloalkyl, preferably C _4-6 cycloalkyl;

d2 A4, 5 or 6 membered saturated or partially unsaturated monocyclic heterocyclyl having 1, 2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur;

d3 C _6-10 aryl; and

D4 A 5 or 6 membered heteroaryl having 1,2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur;

preferably, R ⁴ is H、-CH₃、-CH₂CH₃、-CH₂CH₂CH₃、-CH(CH₃)₂、-CF₃、-CH₂CF₃、-CH₂CH₂CF₃、-CH₂CH₂CH₂CF₃、-CH₂-OH、-CH₂CH₂-OH、-CH₂CH₂CH₂-OH、-C(O)CH₃、-S(O)₂CH₃、
The compound of any one of claims 1-16, wherein R ^x is selected from the following groups:

a) Halogen, -CN, -NO ₂, -OH and-SH;

b) -O-optionally substituted C _1-4 alkyl and-S-optionally substituted C _1-4 alkyl, preferably-O-CH ₃ and-S-CH ₃;

c)-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-NR^3aR^3b、-CONR^3aR^3b and-CO ₂R^3a, wherein R ^3a、R^3b is each as defined in any one of claims 1 to 16, preferably selected from H and optionally substituted C _1-6 alkyl;

d) C _1-6 alkyl which is unsubstituted or substituted by 1,2,3 or more halogen;

preferably, R ^x is selected from F, cl, -CN, CF ₃、-O-CH₃、-SO₂CH₃ or-CO ₂CH₃.
The compound of any one of claims 1-17, wherein R ³ is selected from:
a compound according to any one of claims 1-18, wherein:

r ¹ is-CN;

r ² is hydrogen;

R ³ is Cy;

Cy is as defined in any one of claims 1-10, in particular claims 10-18, preferably selected from 8 to 10 membered saturated or partially unsaturated bicyclic heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, C _6-10 aryl, 5 or 6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur and 8 to 10 membered bicyclic heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the heterocyclyl, aryl and heteroaryl are as defined in any one of claims 1-10, in particular claims 10-18, with respect to Cy, and wherein the heterocyclyl, aryl and heteroaryl are each substituted with 0, 1, 2 or 3R ^x;

R ⁴ is selected from hydrogen, optionally substituted C _1-6 alkyl, and C _4-6 cycloalkyl substituted with 0,1,2, or 3R ^x; and

Each R ^x is independently selected from halogen, -CN, -NO ₂, -OH, -O-optionally substituted C _1-6 alkyl, -SH, -S-optionally substituted C _1-6 alkyl 、-C(O)R^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a、-CONR^3aR^3b, and optionally substituted C _1-6 alkyl;

R ^3a and R ^3b are each independently hydrogen or optionally substituted C _1-6 alkyl.
The compound of any one of claims 1-19, cy is selected from:

Phenyl substituted with 0, 1,2 or 3R ^x;

Naphthyl substituted by 1R ^x;

A 5-or 6-membered heteroaryl substituted with 1 or 2R ^x having 1-3 heteroatoms independently selected from nitrogen, oxygen and sulfur, preferably a 5-membered heteroaryl having 1 nitrogen heteroatom and 0 or 1 heteroatom selected from nitrogen, oxygen and sulfur, more preferably a 6-membered heteroaryl having 1 or 2 nitrogen heteroatoms;

A 9 or 10 membered bicyclic heteroaryl group having 1,2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur, substituted with 1,2 or 3R ^x, wherein the bicyclic heteroaryl group is a fused ring system of a benzene ring and a 5 or 6 membered heteroaryl group, wherein the heteroatoms are all in the 5 or 6 membered heteroaryl ring and are not common atoms; preferably 9 or 10 membered bicyclic heteroaryl groups having 1-2 nitrogen heteroatoms; and

9 Or 10 membered saturated or partially unsaturated bicyclic heterocyclyl substituted with 0, 1,2 or 3R ^x having 1,2 or 3, preferably 1 or 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, wherein the bicyclic heterocyclyl is a fused ring system of a benzene ring and a 5 or 6 membered saturated or partially unsaturated heterocyclyl, wherein the heteroatoms are all in the 5 or 6 membered saturated or partially unsaturated heterocyclyl ring and are not common atoms; preferably a 9 or 10 (especially 9) membered saturated or partially unsaturated bicyclic heterocyclyl having 1 or 2 (especially 1) heteroatoms independently selected from oxygen and sulfur, or a 9 or 10 (especially 9) membered saturated or partially unsaturated bicyclic heterocyclyl having 1 or 2 (especially 1) nitrogen heteroatoms and 0 or 1 (especially 0) oxygen or sulfur heteroatoms;

preferably, cy is selected from:
The compound of any one of claims 1-20, wherein R ⁴ is selected from the following groups:

a)H；

b1 Unsubstituted C _1-6 alkyl, more preferably C _1-3 alkyl;

b2 C _1-4 alkyl substituted with a group selected from: -OH and 1,2,3 or more halogens; and

D1 Unsubstituted C _4-6 cycloalkyl;

Preferably, R ⁴ is H、-CH₃、-CH₂CH₃、-CH₂CH₂CH₃、-CH(CH₃)₂、-CF₃、-CH₂CF₃、-CH₂CH₂CF₃、-CH₂CH₂CH₂CF₃、-CH₂-OH、-CH₂CH₂-OH、-CH₂CH₂CH₂-OH、
The compound of any one of claims 1-21, wherein R ^x is selected from the following groups:

a) Halogen, -CN;

b) -O-optionally substituted C _1-4 alkyl and-S-optionally substituted C _1-4 alkyl, preferably-O-CH ₃ and-S-CH ₃;

c) -C (O) R ^3a、-SO₂R^3a、-SO₂NR^3aR^3b、-CO₂R^3a and-CONR ^3aR^3b, wherein R ^3a、R^3b are each independently H or optionally substituted C _1-6 alkyl, preferably H or optionally substituted C _1-4 alkyl, more preferably H or CH ₃; and

D) C _1-6 alkyl substituted with 1,2, 3 or more halogens;

preferably, R ^x is selected from F, cl, -CN, CF ₃、-O-CH₃、-SO₂CH₃ or-CO ₂CH₃.
The compound of any one of claims 1-22, wherein R ³ is selected from:
The compound of any one of claims 1-23, having a structure represented by one of formulas I-a, I-a-I, I-a-ii, or I-a-iii to I-a-xiv:

Wherein each of R ²、R³ and R ⁴ is as defined in any one of claims 1 to 23;

Wherein each of R ³ and R ⁴ is as defined in any one of the preceding claims 1-23;

Wherein each of Cy and R ⁴ is as defined in any one of claims 1-23;

Wherein each of R ⁴ and R ^x is as defined in any one of claims 1 to 23.
The compound of any one of claims 1-24, having a structure represented by formula I-a-xv:

Wherein R ⁴ is selected from hydrogen and optionally substituted C _1-6 alkyl; and

Each R ^x is independently selected from halogen, -CN and optionally substituted C _1-6 alkyl.
The compound of any one of claims 1-25, wherein Cy is:
The compound of any one of claims 1-26, wherein R ⁴ is selected from the following groups:

a)H；

b1 Unsubstituted C _1-6 alkyl, more preferably C _1-3 alkyl; and

B2 C _1-4 alkyl substituted with 1,2, 3 or more halogens;

Preferably, R ⁴ is H, -CH ₃、-CF₃、-CH₂CF₃、-CH₂CH₂CF₃.
The compound of any one of claims 1-27, wherein R ^x is selected from: halogen; -CN; c _1-6 alkyl substituted with 1,2, 3 or more halogens;

preferably, R ^x is selected from F, cl, -CN and CF ₃.
The compound of any one of claims 1-28, wherein R ³ is selected from:
the compound of claim 1, selected from the following compounds:
a pharmaceutical composition comprising a compound according to any one of claims 1-30, or an enantiomer, diastereomer, racemate, stereoisomer, tautomer, geometric isomer, N-oxide, metabolite, prodrug or pharmaceutically acceptable salt, ester, solvate, hydrate, isotopically labeled compound or prodrug thereof.
A method comprising the steps of:

the compound according to any one of claims 1 to 30 or the pharmaceutical composition according to claim 31 is administered to an individual (i) suffering from a condition characterized by axonal degeneration or (ii) at risk of suffering from a condition characterized by axonal degeneration.
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 30 or a pharmaceutical composition according to claim 31.
A method of treating or preventing a neurodegenerative disease, disorder or condition characterized by axonal degeneration comprising administering to a subject in need thereof a compound of any one of claims 1 to 30 or a pharmaceutical composition of claim 31.
Use of a compound according to any one of claims 1 to 30 or a pharmaceutical composition according to claim 31 in the manufacture of a medicament for the treatment or prevention of axonometric degeneration; or in the manufacture of a medicament for the treatment or prevention of a neurodegenerative disease, disorder or condition characterized by axonal degeneration.
A compound according to any one of claims 1 to 30 or a pharmaceutical composition according to claim 31 for use in the treatment or prevention of axonal degeneration, or for the treatment or prevention of a neurodegenerative disease, disorder or condition characterized by axonal degeneration.
A method of inhibiting SARM1 comprising contacting a biological sample with a compound according to any one of claims 1 to 30 or a pharmaceutical composition according to claim 31.
The method of claim 37, wherein SARM1 is inhibited by covalently modifying a cysteine residue of SARM1.
The method of claim 38, wherein the cysteine residue of SARM1 is Cys635, cys629, or Cys649.