Oxalic acid diamide compound, pharmaceutical composition containing oxalic acid diamide compound, and preparation method and application of oxalic acid diamide compound

Abstract

The present invention relates to an oxalic acid diamide compound represented by formula (I), a pharmaceutical composition comprising the same, a preparation method thereof, and use thereof for the prevention or treatment of diseases or conditions associated with PRMT5 activity. # imgabs0 #

Description

Oxalic acid diamide compound, pharmaceutical composition containing the same, and preparation method and use thereof Technical Field

[0001] The present invention relates to an oxalic acid diamide compound, a pharmaceutical composition containing the same, a preparation method thereof, and use thereof for preventing or treating diseases or conditions associated with PRMT5 activity. Background Art

[0002] PRMT5 (Protein Arginine Methyltransferase 5) is an epigenetic enzyme and a member of the PRMT family (humans have PRMT1-11). It catalyzes the methylation of arginine residues on histones and certain non-histone substrates. PRMT5 is widely present in the nucleus and cytoplasm of human cells, including tissues such as the heart, muscle, and testes. PRMT5 is classified into types I, II, and III based on the mode of arginine methylation catalyzed by it. PRMT5 belongs to the type II symmetric dimethylating (sDMA) PRMT, whose methyl donor is S-adenosylmethionine (SAM).

[0003] PRMT5 regulates the expression of multiple target proteins by catalyzing the arginine methylation of substrates, participating in various physiological functions and playing an important role in tumor cell proliferation, metastasis, and malignant transformation. PRMT5 methylation of histones leads to the silencing of tumor suppressor genes such as p53, ST7, NM23, and Rb, thereby promoting the occurrence and development of tumors. PRMT5 regulation of non-histone proteins is mainly reflected in affecting the localization and expression of transcription factors (NF-κB / P65, E2F1, HoxA\GATA4), programmed cell death protein 4 (PDCD4), cell cycle and survival-related regulatory proteins E2F1, hypoxia-inducible factor 1 (HIF-1), cyclin-dependent kinases (CDKs), PI3K / Akt, etc. (Koh CM, Bezzi M, Guccione E. Curr Mol Bio Rep, 2015, 1(1):19-28). In lung cancer cells, PRMT5 can inhibit the transcription of the miR-99 family, increase FGFR3 expression, activate Erk1 / 2 and Ak pathways, leading to tumor cell growth and metastasis (Pengyu Jing, Nan Zhao, et al. Cancer Letters, 2018, 427, 38-48). In colon cancer, PRMT5 can methylate Eif4e and FGFR3, promoting tumor cell growth (ZHANG B, DONG S, ZHU R, et al. Oncotarget, 2015, 6(26): 22799-22811.).

[0004] MTAP is the gene encoding methylthioadenosine phosphorylase, located on chromosome 9p21, close to the tumor suppressor gene CDKN2A (which often undergoes homozygous deletion). Therefore, MTAP is often co-deleted with CDKN2A in tumors (Marjon K, Kalev P, Marks K. Annual Review of Cancer Biology, 2021, 5(1)). Approximately 15% of solid tumors suffer from MTAP deletion.

[0005] PRMT5 has two cofactors: an activating cofactor (SAM) and an inhibitory cofactor (MTA; 5'-methylthioadenosine). In normal cells, MTAP is responsible for converting MTA to Met (methionine), while PRMT5 is responsible for converting SAM to SAH (S-adenosyl-L-homocysteine). MTA is an endogenous competitive inhibitor of PRMT5-SAM. In tumor cells, the loss of MTAP leads to the accumulation of MTA, partially inhibiting PRMT5 activity and making tumor cells more dependent on PRMT5. In these cases, inhibiting PRMT5 can further block the methylation function of PRMT5, causing tumor cell death.

[0006] In summary, inhibition of PRMT5 exhibits synthetic lethality in the absence of MTAP. Currently, no PRMT5 inhibitors are available. Therefore, the development of novel, highly effective, and low-toxic PRMT5 inhibitors is needed to meet clinical needs.

[0007] Summary of the Invention

[0008] The present invention provides a new oxalic acid diamide compound, which has a good inhibitory effect on MTAP-deficient tumor cells and has good pharmacokinetic properties.

[0009] One aspect of the present invention provides a compound of Formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof:

[0010] in:

[0011] represents a single bond or a double bond;

[0012] X 1 、X 2 and X 3 are each independently selected from -CH2-, -CH-, -O-, -N-, -NH- and -S-;

[0013] R 1Each occurrence is independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl;

[0014] Ring A is a 5-7 membered saturated or partially saturated heterocyclic ring containing at least one N atom and optionally 1 or 2 other heteroatoms, each independently selected from O or S, which may be the same or different;

[0015] R 2 is independently selected at each occurrence from H, -OH, halogen, -CN, -NR 4 R 5 、-COCH3、-NHCOCH3、C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Hydroxyalkyl, C 2-6 Heteroalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, C 3-8 Cycloalkoxy, C 6-10 Aryl and 5-10 membered heteroaryl, said alkyl, alkoxy, hydroxyalkyl, heteroalkyl, cycloalkyl, heterocyclyl, cycloalkoxy, aryl or heteroaryl being each optionally substituted by one or more R 3 replace;

[0016] R 3 is independently selected at each occurrence from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, C 3-6 Cycloalkyl, C 3-8 Cycloalkoxy, 3-8 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl, wherein the alkyl, alkoxy, heteroalkyl, cycloalkyl, cycloalkoxy, heterocyclyl, aryl or heteroaryl are each optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 3-8 Substitution of cycloalkoxy and 3-8 membered heterocyclic groups;

[0017] R 4 、R 5 、R 6 and R 7 Each independently selected from H and C 1-6 Alkyl; or

[0018] R 4 and R 5 、R 6 and R 7 Together with the nitrogen atom to which it is attached, it forms a 3-8 membered heterocyclic group, wherein the heterocyclic group is optionally substituted by one or more independently selected from halogen, OH, CN, -NH2 and C 1-6 Substitution of alkyl groups;

[0019] m is 0, 1, 2, or 3;

[0020] n is 0, 1, 2, 3 or 4.

[0021] Another aspect of the present invention provides a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof and one or more pharmaceutically acceptable carriers.

[0022] Another aspect of the present invention provides the use of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, or a pharmaceutical composition of the present invention in the preparation of a medicament for preventing or treating a disease or condition associated with PRMT5 activity.

[0023] Another aspect of the present invention provides a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, or a pharmaceutical composition of the present invention, for use in preventing or treating a disease or condition associated with PRMT5 activity.

[0024] Another aspect of the present invention provides a method for preventing or treating a disease or condition associated with PRMT5 activity, comprising administering to a subject in need thereof an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, or a pharmaceutical composition of the present invention.

[0025] Another aspect of the present invention provides a method for preparing a compound of the present invention, comprising the steps of:

[0026] Step 1: Compound IA-1 and acyl chloride Compound IA-2 is generated through condensation reaction;

[0027] Step 2: Compound IA-2 is hydrolyzed to generate compound IA-3;

[0028] Step 3: Compound IA-3 and compound IA-4 undergo condensation reaction to produce compound IA-5;

[0029] Step 4: The condensation product IA-5 is deprotected to generate compound I;

[0030] in

[0031] PG is an amino protecting group; and

[0032] X 1 、X 2 、X 3 , Ring A, R 1 、R 2 , m, n are as defined above, R 6 and R 7 For H.

[0033] definition

[0034] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as those commonly understood by those skilled in the art. References to technology used herein are intended to refer to technology commonly understood in the art, including variations of technology or substitutions of equivalent technology that would be apparent to those skilled in the art. While it is believed that the following terms are well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

[0035] The terms "comprising," "including," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps, even though the additional unrecited elements or method steps are not necessarily present (i.e., these terms also encompass the terms "consisting essentially of" and "consisting of.").

[0036] As used herein, the term "alkyl" is defined as a linear or branched saturated aliphatic hydrocarbon. In some embodiments, the alkyl group has 1 to 12, such as 1 to 6, 1 to 4 carbon atoms. For example, as used herein, the term "C 1-6 Alkyl" and "C 1-4"Alkyl" refers to a linear or branched group having 1 to 6 carbon atoms and 1 to 4 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl or n-hexyl), which is optionally substituted by one or more (e.g., 1 to 3) suitable substituents such as halogen (in which case the group is referred to as "haloalkyl") (e.g., CH2F, CHF2, CF3, CCl3, C2F5, C2Cl5, CH2CF3, CH2Cl or -CH2CH2CF3, etc.). The term "C 1-4 "Alkyl" refers to a linear or branched aliphatic hydrocarbon chain having from 1 to 4 carbon atoms (ie, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl).

[0037] As used herein, the term "heteroalkyl" refers to an alkyl group having one or more backbone atoms independently selected from atoms other than carbon, such as oxygen, nitrogen, sulfur, phosphorus, or a combination thereof, in the backbone carbon atoms of the alkyl group. Numerical ranges (e.g., C 2-6 Assorted alkyl) refers to the number of carbons in the chain, which in this example includes 2-6 carbon atoms. For example, a -CH2OCH2CH3 group is referred to as a C3 assorted alkyl, and a -CH2OCH2CH2NHCH3 group is referred to as a C4 assorted alkyl. Connection to the rest of the molecule can be through heteroatoms or carbon atoms in the assorted alkyl chain.

[0038] As used herein, the term "haloalkyl" refers to an alkyl group substituted by one or more (such as 1 to 3) the same or different halogen atoms. 1-8 Halogenated alkyl, "C 1-6 Haloalkyl" and "C 1-4 The term "haloalkyl" refers to a haloalkyl group having 1 to 8 carbon atoms, 1 to 6 carbon atoms, and 1-4 carbon atoms, respectively, such as -CF3, -C2F5, -CHF2, -CH2F, -CH2CF3, -CH2Cl, or -CH2CH2CF3.

[0039] As used herein, the term "hydroxyalkyl" refers to a group in which a hydrogen atom in an alkyl group is replaced by one or more hydroxyl groups, for example, C 1-6 Hydroxyalkyl or C 1-4 Examples of hydroxyalkyl include, but are not limited to, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, -CH(OH)CH3, and the like.

[0040] As used herein, the term "alkoxy" refers to an -O-alkyl group, wherein alkyl is as defined above, for example, C 1-8 Alkoxy, C 1-6 Alkoxy, C 1-4 Alkoxy or C 1-3 Alkoxy. C 1-6Representative examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, and the like, wherein the alkoxy groups are optionally substituted with one or more (such as 1 to 3) identical or different substituents. For example, the term "haloalkoxy" refers to an alkoxy group wherein the hydrogen atoms are substituted with one or more (such as 1 to 3) identical or different halogen atoms.

[0041] As used herein, the term "paracyclic" or "fused ring" refers to a ring system formed by two or more cyclic structures that share two adjacent atoms.

[0042] As used herein, the term "spirocycle" refers to a ring system formed by two or more cyclic structures that share one ring atom with each other.

[0043] As used herein, the term "bridged ring" refers to a ring system formed by two or more cyclic structures sharing two atoms that are not directly connected to each other.

[0044] As used herein, the term "cycloalkyl" refers to a saturated or unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring group, including but not limited to monocyclic alkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, etc.) and bicyclic alkyl, including spirocyclic, annular (condensed) or bridged ring systems (i.e., spirocyclic alkyl, annular (condensed) alkyl and bridged cycloalkyl, such as bicyclo [1.1.1] pentyl, bicyclo [2.2.1] heptyl, etc.). In the present invention, cycloalkyl is optionally substituted with one or more (such as 1 to 3) identical or different substituents. The carbon atoms on the cycloalkyl are optionally substituted with oxo (oxo) groups (i.e., forming C=O). The term "C 3-8 "Cycloalkyl" refers to a cycloalkyl group having 3 to 8 ring carbon atoms, such as C 3-6 Cycloalkyl, which may be a monocyclic alkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl, or a bicyclic alkyl, such as C 5-8 Spiroalkyl, C 5-8 Bridged cycloalkyl, C 5-8 Condensed cycloalkyl, C 5-6 Spiroalkyl, C 5-6 Bridged cycloalkyl or C 5-6 Fused cycloalkyl.

[0045] As used herein, the term "cycloalkoxy" refers to -O-cycloalkyl, wherein cycloalkyl is as defined above. Representative examples of cycloalkoxy include, but are not limited to, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.

[0046] As used herein, the term "heterocyclyl" or "heterocycle" refers to an aliphatic monocyclic or polycyclic (e.g., cyclic, spirocyclic or bridged) group having 2 or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14) carbon atoms and one or more (e.g., 1, 2, 3 or 4) heteroatoms, including but not limited to oxygen atoms, nitrogen atoms and sulfur atoms, wherein the carbon atoms and heteroatoms on the heterocyclyl are optionally substituted with oxo groups (e.g., forming C=O, S(=O) or S(=O)2), or are optionally substituted with one or more (e.g., 1 to 3) independently selected from halogen and C 1-3 The term "saturated heterocycle" refers to a fully saturated heterocycle, such as a tetrahydrofuran ring, a piperidine ring, a morpholine ring, a tetrahydropyran ring, a piperazine ring, etc. The term "partially saturated heterocycle" refers to a heterocycle containing both saturated single bonds and unsaturated double bonds, such as 3,4-dihydro-2H-pyran, 1,2,3,4-tetrahydropyridine, 4,5-dihydroisoxazolyl, 4,5-dihydrooxazolyl, 2,5-dihydrooxazolyl, 2,3-dihydrooxazolyl, etc. As used herein, the term "4-11 membered heterocyclyl" means a heterocyclyl containing 4-11 ring atoms, including but not limited to 4-10 membered heterocyclyl, 4-9 membered heterocyclyl, 4-8 membered heterocyclyl, 4-7 membered heterocyclyl, 5-6 membered heterocyclyl, 3-8 membered heterocyclyl, 3-7 membered heterocyclyl, 4-7 membered nitrogen-containing heterocyclyl, 4-7 membered oxygen-containing heterocyclyl, 4-7 membered sulfur-containing heterocyclyl, 5-6 membered nitrogen-containing heterocyclyl, 5-6 membered oxygen-containing heterocyclyl, 5-6 membered sulfur-containing heterocyclyl, etc., wherein the "nitrogen-containing heterocyclyl", "oxygen-containing heterocyclyl" and "sulfur-containing heterocyclyl" each optionally further contain one or more other heteroatoms independently selected from oxygen, nitrogen and sulfur. Examples of 4-11 membered heterocyclyls include but are not limited to oxiranyl, aziridine, azetidinyl, oxetanyl, tetrahydrofuranyl, pyrrolidinyl, pyrrolidonyl (such as ), imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl. As used herein, the term "3-8 membered heterocyclyl" means a heterocyclyl containing 3-8 ring atoms, including but not limited to a 3-8 membered heterocyclyl, a 3-7 membered heterocyclyl, a 3-6 membered heterocyclyl, a 4-8 membered heterocyclyl, a 4-7 membered heterocyclyl, a 4-6 membered heterocyclyl, a 5-6 membered heterocyclyl, a 4-7 membered nitrogen-containing heterocyclyl, a 4-7 membered oxygen-containing heterocyclyl, a 4-7 membered sulfur-containing heterocyclyl, a 5-6 membered nitrogen-containing heterocyclyl, a 5-6 membered oxygen-containing heterocyclyl, a 5-6 membered sulfur-containing heterocyclyl, etc., and each of the "nitrogen-containing heterocyclyl", "oxygen-containing heterocyclyl" and "sulfur-containing heterocyclyl" optionally further contains one or more other heteroatoms independently selected from oxygen, nitrogen and sulfur. Examples of 3-8 membered heterocyclic groups include, but are not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, pyrrolidinyl, pyrrolidonyl (e.g. ), imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl.

[0047] In the present invention, the heterocyclic group can form a parallel ring structure with a heterocyclic group or a cycloalkyl group, and the connection point of the parallel ring structure with the other groups can be on any heterocyclic group or cycloalkyl group. Therefore, the heterocyclic group of the present invention also includes (but is not limited to) heterocyclic and heterocyclic groups, heterocyclic and cycloalkyl groups, monoheterocyclic and monoheterocyclic groups, and monoheterocyclic and monocycloalkyl groups, such as 3-7 membered (mono) heterocyclic groups and 3-7 membered (mono) heterocyclic groups, 3-7 membered (mono) heterocyclic groups and (mono) cycloalkyl groups, 3-7 membered (mono) heterocyclic groups and C 4-6 (Mono)cycloalkyl, examples of which are not limited to pyrrolidinyl and cyclopropyl, cyclopentyl and aziridine, pyrrolidinyl and cyclobutyl, pyrrolidinyl and pyrrolidinyl, pyrrolidinyl and piperidinyl, pyrrolidinyl and piperazinyl, piperidinyl and morpholinyl,

[0048] In the present invention, the heterocyclic group also includes a bridged heterocyclic group and a spiro heterocyclic group.

[0049] As used herein, the term "bridged heterocycle" refers to a cyclic structure containing one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen atoms, nitrogen atoms, and / or sulfur atoms) formed by two saturated rings sharing two ring atoms that are not directly connected, including but not limited to 7-10 membered bridged heterocycles, 8-10 membered bridged heterocycles, 7-10 membered nitrogen-containing bridged heterocycles, 7-10 membered oxygen-containing bridged heterocycles, 7-10 membered sulfur-containing bridged heterocycles, etc., for example The “nitrogen-containing bridged heterocycle”, “oxygen-containing bridged heterocycle” and “sulfur-containing bridged heterocycle” optionally further contain one or more other heteroatoms independently selected from oxygen, nitrogen and sulfur.

[0050] As used herein, the term "spiroheterocycle" refers to a cyclic structure containing one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen atoms, nitrogen atoms, sulfur atoms) formed by two or more saturated rings sharing a ring atom, including but not limited to 5-10 membered spiroheterocycles, 6-10 membered spiroheterocycles, 6-10 membered nitrogen-containing spiroheterocycles, 6-10 membered oxygen-containing spiroheterocycles, 6-10 membered sulfur-containing spiroheterocycles, etc., for example The "nitrogen-containing spiroheterocycle", "oxygen-containing spiroheterocycle" and "sulfur-containing spiroheterocycle" optionally further contain one or more other heteroatoms independently selected from oxygen, nitrogen and sulfur. The term "6-10 membered nitrogen-containing spiroheterocyclyl" refers to a spiroheterocyclyl containing a total of 6-10 ring atoms, at least one of which is a nitrogen atom.

[0051] Examples of the group obtained by condensing a heterocyclic group with an aryl group include, but are not limited to:

[0052] As used herein, the term "aryl", "phenyl" or "aromatic ring" refers to an all-carbon monocyclic or fused polycyclic aromatic group having a conjugated π electron system. 6-10 "Aryl (aromatic ring)" means an aromatic group (aromatic ring) containing 6 to 10 carbon atoms, preferably a phenyl (phenyl ring) or a naphthyl (naphthalene ring). The aryl group is optionally substituted by one or more (such as 1 to 3) identical or different substituents (e.g., halogen, OH, CN, NO2, C1-C6 alkyl, etc.).

[0053] As used herein, the term "heteroaryl" or "heteroaromatic ring" refers to a monocyclic or polycyclic aromatic group containing one or more identical or different heteroatoms, including monocyclic heteroaryl groups and bicyclic or polycyclic ring systems containing at least one heteroaromatic ring (an aromatic ring system containing at least one heteroatom), which can have 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, for example 5, 6, 7, 8, 9 or 10 ring atoms, and in each case can be benzo-fused. The heteroatom can be oxygen, nitrogen or sulfur. The carbon atoms and heteroatoms on the heteroaryl group are optionally substituted with oxo groups (for example, to form C=O, S(=O) or S(=O)2).

[0054] As used herein, the term "5-10 membered heteroaryl" or "5-10 membered heteroaromatic ring" means a heteroaryl group (heteroaromatic ring) containing 5 to 10 (e.g., 5 to 6) ring atoms, including a 5-10 membered nitrogen-containing heteroaryl group, a 5-10 membered oxygen-containing heteroaryl group, a 5-10 membered sulfur-containing heteroaryl group, a 5-6 membered nitrogen-containing heteroaryl group, a 5-6 membered oxygen-containing heteroaryl group, a 5-6 membered sulfur-containing heteroaryl group, etc. The "nitrogen-containing heteroaryl group," "oxygen-containing heteroaryl group," and "sulfur-containing heteroaryl group" each optionally contain one or more other heteroatoms independently selected from oxygen, nitrogen, and sulfur. Examples include, but are not limited to, thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, triazolyl, tetrazolyl, oxadiazolyl, thiadiazolyl, etc., or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., as well as 5-10 membered cyclic groups containing these groups.

[0055] In the present invention, a heteroaryl group (e.g., a monoheteroaryl group) can share two adjacent atoms with an aryl group (e.g., a monocyclic aryl group, such as a phenyl group), a heterocyclic group (e.g., a monoheterocyclic group), a cycloalkyl group (e.g., a monocycloalkyl group) or another heteroaryl group (e.g., another monoheteroaryl group) to form a parallel ring structure, and the connection point can be on any heteroaryl ring or on other rings, including but not limited to (mono)heteroaryl and (mono)heteroaryl, (mono)heteroaryl and (mono)heterocyclic group and (mono)heteroaryl and (mono)cycloalkyl, such as a 5-6 membered (mono)heteroaryl and 5-6 membered (mono)heteroaryl, a 5-6 membered (mono)heteroarylphenyl group, a 5-6 membered (mono)heteroaryl and 5-6 membered (mono)heterocyclic group or a 5-6 membered (mono)heteroaryl and C 4-6 (mono)cycloalkyl (e.g., 5-6 membered heteroarylcyclobutyl, 5-6 membered heteroarylcyclopentyl or 5-6 membered heteroarylcyclohexyl), examples of which are not limited to benzothiazolyl, indolyl, isoindolyl, indazolyl, benzimidazole, quinolinyl, isoquinolinyl, wait.

[0056] As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

[0057] The term "substituted" means that one or more (e.g., one, two, three, or four) hydrogen atoms on the designated atom are replaced with a group selected from the indicated group, provided that the designated atom's normal valence is not exceeded in the current context and that the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0058] If a substituent is described as being "optionally substituted with one or more...", the substituent may be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more of the listed substituents, one or more hydrogens on the carbon (to the extent of any hydrogens present) may be replaced, individually and / or collectively, with independently selected optional substituents. If a nitrogen of a substituent is described as being optionally substituted with one or more of the listed substituents, one or more hydrogens on the nitrogen (to the extent of any hydrogens present) may each be replaced with an independently selected optional substituent.

[0059] If substituents are described as being "independently selected" from a group, each substituent is selected independently of the other. Thus, each substituent may be the same as or different from another (other) substituent.

[0060] As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5 or 10, where reasonable.

[0061] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0062] When a bond to a substituent is shown to pass through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring atom in the substitutable ring.

[0063] The present invention also includes all pharmaceutically acceptable isotopically labeled compounds, which are identical to the compounds 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 prevalent in nature. Examples of isotopes suitable for inclusion in the compounds of the present invention include, but are not limited to, isotopes of hydrogen (e.g., deuterium ( 2 H), tritium ( 3 H)); carbon isotopes (e.g. 11 C. 13 C and 14 C); isotopes of chlorine (e.g. 36 Cl); isotopes of fluorine (e.g. 18 F); isotopes of iodine (such as 123 I and 125 I); isotopes of nitrogen (e.g. 13 N and 15 N); oxygen isotopes (e.g. 15 O. 17 O and 18 O); isotopes of phosphorus (such as 32 P); and sulfur isotopes (e.g. 35S). Certain isotopically labeled compounds of the invention (e.g., those incorporating radioactive isotopes) are useful in drug and / or substrate tissue distribution studies (e.g., assays). The radioactive isotope tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) are particularly useful for this purpose because they are easy to incorporate and easy to detect. 11 C. 18 F. 15 O and 13 N) substitution can be used to examine substrate receptor occupancy in positron emission tomography (PET) studies. Isotopically labeled compounds of the present invention can be prepared by methods analogous to those described in the accompanying schemes and / or examples and preparations by using appropriate isotopically labeled reagents instead of the non-labeled reagents previously employed. Pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent is isotopically substituted, for example, D2O, acetone-d6 or DMSO-d6.

[0064] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In a compound with one or more (e.g., one, two, three, or four) asymmetric centers, it can produce a racemic mixture, a single enantiomer, a diastereomeric mixture, and a separate diastereomer. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. For example, nitroso-oxime can exist in the following tautomeric form equilibrium in solution:

[0065] It is to be understood that the scope of this application encompasses all such isomers in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%) or mixtures thereof.

[0066] In this article, solid lines can be used Solid wedge or virtual wedge The chemical bonds of the compounds of the present invention are depicted. The use of solid lines to depict bonds to asymmetric carbon atoms is intended to indicate that all possible stereoisomers at that carbon atom are included (e.g., specific enantiomers, racemic mixtures, etc.). The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the indicated stereoisomers exist. When present in a racemic mixture, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the present invention are intended to exist as stereoisomers, including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, atropisomers, and mixtures thereof. The compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

[0067] 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 more than one polymorph in any ratio.

[0068] A cocrystal refers to a drug active molecule and other physiologically acceptable acid, base, salt, or non-ionic compound molecules bound in the same crystal lattice by hydrogen bonds, π-π stacking, van der Waals forces, and other non-covalent bonds.

[0069] It should also be understood that certain compounds of the present invention may be used therapeutically in free form 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, N-oxides, metabolites, or prodrugs that, upon administration to a patient in need thereof, are capable of directly or indirectly providing a compound of the present invention or a metabolite or residue thereof. Therefore, when reference is made herein to a "compound of the present invention," such various derivative forms of the compound are also intended to be encompassed.

[0070] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.

[0071] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof, such as hexafluorophosphate salts and meglumine salts. For a review of suitable salts, see Stahl and Wermuth, "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley-VCH, 2002).

[0072] As used herein, the term "ester" refers to esters derived from the compounds of the general formulae herein, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the present invention in the form of free acid or alcohol). The compounds of the present invention themselves may also be esters.

[0073] The compounds of the present invention may exist in the form of solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0074] Those skilled in the art will appreciate that, because nitrogen requires available lone pairs of electrons to be oxidized to oxides, not all nitrogen-containing heterocycles can form N-oxides. Those skilled in the art will recognize nitrogen-containing heterocycles that can form N-oxides. Those skilled in the art will also recognize that tertiary amines can form N-oxides. The synthetic method for preparing the N-oxide of heterocycles and tertiary amines is well known to those skilled in the art, including but not limited to oxidizing heterocycles and tertiary amines with peroxyacids such as Peracetic Acid and Metachloroperbenzoic Acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate and dioxirane such as dimethyldioxirane. These methods for preparing N-oxides have been extensively described and reviewed in the literature, see for example: TL Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp 748-750; AR Katritzky and AJ Boulton, Eds., Academic Press; and GWH Cheeseman and ESGWerstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp 390-392, AR Katritzky and AJ Boulton, Eds., Academic Press.

[0075] Also included within the scope of the present invention are metabolites of the compounds of the invention, i.e., substances formed in vivo upon administration of the compounds of the invention. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc. of the administered compound. Thus, the present invention includes metabolites of the compounds of the invention, including compounds produced by contacting a compound of the invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

[0076] The present invention further includes within its scope prodrugs of the compounds of the present invention, which are certain derivatives of the compounds of the present invention that may themselves have little or no pharmacological activity and can be converted into compounds of the present invention having the desired activity by, for example, hydrolytic cleavage when administered to the body or thereon. Typically, such prodrugs will be functional group derivatives of the compounds that are readily converted into the desired therapeutically active compounds in vivo. Additional information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems", Volume 14, ACS Symposium Series (T. Higuchi and V. Stella). Prodrugs of the present invention can be prepared, for example, by replacing appropriate functional groups present in the compounds of the present invention with certain moieties known to those skilled in the art as "pro-moieties" (e.g., as described in "Design of Prodrugs", H. Bundgaard (Elsevier, 1985)).

[0077] The present invention also encompasses compounds of the present invention that contain protecting groups. During any process for preparing the compounds of the present invention, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules involved, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved using conventional protecting groups, for example, those described in TW Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

[0078] The term "about" means within ±10%, preferably within ±5%, and more preferably within ±2% of the stated numerical value.

[0079] Compound

[0080] In some embodiments, the present invention provides a compound of Formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite, or prodrug thereof:

[0081] in:

[0082] represents a single bond or a double bond;

[0083] X 1 、X 2 and X 3are each independently selected from -CH2-, -CH-, -O-, -N-, -NH- and -S-;

[0084] R 1 Each occurrence is independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl;

[0085] Ring A is a 5-7 membered saturated or partially saturated heterocyclic ring containing at least one N atom and optionally 1 or 2 other heteroatoms, each independently selected from O or S, which may be the same or different;

[0086] R 2 is independently selected at each occurrence from H, -OH, halogen, -CN, -NR 4 R 5 、-COCH3、-NHCOCH3、C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Hydroxyalkyl, C 2-6 Heteroalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, C 3-8 Cycloalkoxy, C 6-10 Aryl and 5-10 membered heteroaryl, said alkyl, alkoxy, hydroxyalkyl, heteroalkyl, cycloalkyl, heterocyclyl, cycloalkoxy, aryl or heteroaryl being each optionally substituted by one or more R 3 replace;

[0087] R 3 is independently selected at each occurrence from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, C 3-6 Cycloalkyl, C 3-8 Cycloalkoxy, 3-8 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl, wherein the alkyl, alkoxy, heteroalkyl, cycloalkyl, cycloalkoxy, heterocyclyl, aryl or heteroaryl are each optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 3-8 Substitution of cycloalkoxy and 3-8 membered heterocyclic groups;

[0088] R 4 、R 5 、R 6 and R 7 Each independently selected from H and C 1-6 Alkyl; or

[0089] R 4 and R 5 、R 6 and R 7 Together with the nitrogen atom to which it is attached, it forms a 3-8 membered heterocyclic group, wherein the heterocyclic group is optionally substituted by one or more independently selected from halogen, OH, CN, -NH2 and C 1-6 Substitution of alkyl groups;

[0090] m is 0, 1, 2, or 3;

[0091] n is 0, 1, 2, 3 or 4.

[0092] In certain embodiments, the present invention provides compounds of formula I, wherein X 1 、X 2 and X 3 are each independently selected from -CH2-, -CH-, -O-, -N- and -NH-.

[0093] In certain embodiments, the present invention provides compounds of formula I, wherein X 1 is -CH-, and X 2 and X 3 For -N-.

[0094] In certain embodiments, the present invention provides compounds of formula I, wherein X 1 and X 2 is -N-, and X 3 It is -CH-.

[0095] In certain embodiments, the present invention provides compounds of formula I, wherein X 1 and X 3 is -CH2-, and X 2 It is -O-.

[0096] In certain embodiments, the present invention provides compounds of formula I, wherein R 1 Each occurrence is independently selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-8 Cycloalkyl.

[0097] In certain embodiments, the present invention provides compounds of formula I, wherein R 1 Each occurrence is independently selected from H and C 1-6 alkyl.

[0098] In certain embodiments, the present invention provides compounds of formula I, wherein R 1 It is a methyl group.

[0099] In certain embodiments, in the compound of formula I provided by the present invention, Ring A is a 5-7 membered saturated heterocyclic ring containing at least one N atom and optionally one or two other heteroatoms independently selected from O or S, which may be the same or different.

[0100] In certain embodiments, the present invention provides compounds of formula I, wherein ring A is a 5-7 membered saturated nitrogen heterocycle and a 5-7 membered saturated N- and O-containing heterocycle;

[0101] In certain embodiments, the present invention provides compounds of formula I, wherein ring A is a 6-membered saturated nitrogen heterocycle and a 6-membered saturated N- and O-containing heterocycle;

[0102] In certain embodiments, in the compound of formula I provided by the present invention, ring A is a piperidine ring or a morpholine ring.

[0103] In certain embodiments, in the compound of formula I provided by the present invention, Ring A is a 5-7 membered saturated nitrogen heterocycle.

[0104] In certain embodiments, the present invention provides compounds of formula I, wherein Ring A is a 6-membered saturated nitrogen heterocycle.

[0105] In certain embodiments, the present invention provides compounds of formula I wherein ring A is a piperidine ring.

[0106] In certain embodiments, in the compound of formula I provided by the present invention, Ring A is a morpholine ring.

[0107] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 Each occurrence is independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Hydroxyalkyl, C 2-6 Heteroalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, C 3-8 Cycloalkoxy, C 6-10 Aryl and 5-10 membered heteroaryl, said alkyl, alkoxy, hydroxyalkyl, heteroalkyl, cycloalkyl, heterocyclyl, cycloalkoxy, aryl or heteroaryl being each optionally substituted by one or more R 3 replace.

[0108] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 Each occurrence is independently selected from H, halogen, C 1-6 Alkyl, C 6-10 Aryl and 5-10 membered heteroaryl, said alkyl, aryl or heteroaryl being each optionally substituted by one or more R 3 replace.

[0109] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 Each occurrence is independently selected from C 1-6 Alkyl, C 6-10 Aryl and 6-9 membered heteroaryl, said alkyl, aryl or heteroaryl being each optionally substituted by one or more R 3 replace.

[0110] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 Each occurrence is independently selected from C 1-6 alkyl, phenyl, pyridyl and benzothiazolyl, said phenyl, pyridyl or benzothiazolyl being each optionally substituted with one or more R 3 replace.

[0111] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 Each occurrence is independently selected from C 1-6 Alkyl and 6-9 membered heteroaryl, said alkyl or heteroaryl each being optionally substituted by one or more R 3 replace.

[0112] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 Each occurrence is independently selected from C 1-6 alkyl, pyridyl and benzothiazolyl, said pyridyl or benzothiazolyl being each optionally substituted with one or more R 3 replace.

[0113] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 Each occurrence is independently selected from C 1-6 alkyl, pyridyl and benzothiazolyl, said pyridyl or benzothiazolyl being each optionally substituted by C 1-6 Alkyl, C 1-6 and 5-6 membered heterocyclic groups, the heterocyclic groups are optionally substituted by C 1-6 Alkyl substitution.

[0114] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 is independently selected at each occurrence from methyl, The wavy line represents the point of attachment of the group to the rest of the molecule.

[0115] In certain embodiments, the present invention provides compounds of formula I, wherein R 3 is independently selected at each occurrence from halogen, -CN, -NR 4 R 5 、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, 3-8 membered heterocyclic group, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the alkyl, heteroalkyl, heterocyclyl, aryl or heteroaryl are each optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 3-8 cycloalkoxy and 3-8 membered heterocyclic groups.

[0116] In certain embodiments, the present invention provides compounds of formula I, wherein R 3 is independently selected at each occurrence from halogen, -NR 4 R 5 、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 2-6 heteroalkyl, 4-7 membered heterocyclyl, phenyl and 5-6 membered heteroaryl, wherein each of the alkyl, heteroalkyl, heterocyclyl, phenyl or heteroaryl is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 The substituents of the cycloalkyl group and the 4- to 6-membered heterocyclic group are substituted.

[0117] In certain embodiments, the present invention provides compounds of formula I, wherein R 3 Each occurrence is independently selected from -NR 4 R 5 、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C1-6 Halogenated alkoxy, 4-7 membered heterocyclic group and 5-6 membered heteroaryl, the alkyl, heterocyclic group, heteroaryl are optionally substituted with one or more -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 The substituents of the cycloalkyl group and the 4- to 6-membered heterocyclic group are substituted.

[0118] In certain embodiments, the present invention provides compounds of formula I, wherein R 3 is independently selected at each occurrence from halogen, -NR 4 R 5 、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Heteroalkyl, 5-6 membered heterocyclyl, phenyl and 5-6 membered heteroaryl, wherein each of the alkyl, heteroalkyl, heterocyclyl, phenyl or heteroaryl is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 3-6 Cycloalkyl and C 1-6 The substituents of the haloalkyl group are substituted.

[0119] In certain embodiments, the present invention provides compounds of formula I, wherein R 3 Each occurrence is independently selected from C 1-6 Alkyl, C 1-6 Haloalkyl and 5-6 membered heterocyclic groups, the heterocyclic groups are optionally substituted by one or more C 1-6 Alkyl substitution.

[0120] In certain embodiments, the present invention provides compounds of formula I, wherein R 3 Each occurrence is independently selected from methyl, trifluoromethyl, ethyl, amino, methylamino, trifluoromethoxy, The wavy line represents the point of attachment of the group to the rest of the molecule.

[0121] In certain embodiments, the present invention provides compounds of formula I, wherein R 4 and R 5 are each independently selected from H and methyl.

[0122] In certain embodiments, the present invention provides compounds of formula I, wherein R 6 and R 7 are each independently selected from H and methyl.

[0123] In certain embodiments, the present invention provides compounds of formula I, wherein R 6 and R 7 For H.

[0124] In certain embodiments, the present invention provides compounds of formula I, wherein m is 0 or 1.

[0125] In certain embodiments, the present invention provides compounds of formula I wherein n is 2.

[0126] In certain embodiments, the compound of formula I provided herein is a compound of formula II:

[0127] where X 1 、X 2 、X 3 、R 1 、R 3 , m are as defined above for the compound of formula I.

[0128] In certain embodiments, in the compound of Formula II,

[0129] X 1 is -CH-, and X 2 and X 3 is -N-;

[0130] R 1 is methyl; and

[0131] m is 1.

[0132] In certain embodiments, the compound of formula I provided herein is a compound of formula III:

[0133] where X 1 、X 2 、X 3 、R 1 、R 3 , m are as defined above for the compound of formula I.

[0134] In certain embodiments, in the compound of Formula III,

[0135] X 1 is -CH-, and X 2 and X 3 is -N-;

[0136] R 1 is methyl;

[0137] m is 1; and

[0138] R 3Selected from methyl, trifluoromethyl, ethyl, amino, methylamino, trifluoromethoxy, The wavy line represents the point of attachment of the group to the rest of the molecule.

[0139] In certain embodiments, the compound of formula I provided herein is a compound of formula IV:

[0140] where X 1 、X 2 、X 3 、R 1 、R 3 , m are as defined above for the compound of formula I.

[0141] In certain embodiments, in the compound of Formula IV,

[0142] X 1 is -CH-, and X 2 and X 3 is -N-;

[0143] R 1 is methyl;

[0144] m is 1; and

[0145] R 3 Selected from methyl, trifluoromethyl, ethyl, amino, methylamino, trifluoromethoxy,

[0146] The wavy line represents the point of attachment of the group to the rest of the molecule.

[0147] In certain embodiments, the compound of formula I provided herein is a compound of formula V:

[0148] where X 1 、X 2 、X 3 、R 1 、R 3 , m is as defined above for the compound of formula I; Y 1 、Y 2 are independently selected from N and CH.

[0149] In certain embodiments, the compound of formula V provided herein is a compound of formula V-1:

[0150] where R 3 As defined above for compounds of formula I; Y1 、Y 2 are independently selected from N and CH.

[0151] In certain embodiments, the compound of formula V-1 provided herein is a compound of formula V-1a:

[0152] where R 3 As defined above for the compounds of formula I.

[0153] In certain embodiments, the compound of formula V-1 provided herein is a compound of formula V-1b:

[0154] where R 3 As defined above for the compounds of formula I.

[0155] In certain embodiments, the compound of formula V-1 provided herein is a compound of formula V-1c:

[0156] where R 3 As defined above for the compounds of formula I.

[0157] In certain embodiments, in the compound of Formula V, V-1, V-1a, V-1b or V-1c,

[0158] R 3 Selected from halogen, -CN, -NR 4 R 5 、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, 3-8 membered heterocyclic group, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the alkyl, heteroalkyl, heterocyclyl, aryl or heteroaryl are each optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 3-8 Substitution of cycloalkoxy and 3-8 membered heterocyclic groups;

[0159] Preferably, R 3 Selected from halogen, -NR 4 R 5 、C 1-6 Alkyl, C1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 2-6 heteroalkyl, 4-7 membered heterocyclyl, phenyl and 5-6 membered heteroaryl, wherein each of the alkyl, heteroalkyl, heterocyclyl, phenyl or heteroaryl is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Substitution of cycloalkyl and 4-6 membered heterocyclic groups;

[0160] Preferably, R 3 Selected from-NR 4 R 5 、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, 4-7 membered heterocyclic group and 5-6 membered heteroaryl, the alkyl, heterocyclic group, heteroaryl are optionally substituted with one or more -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Substitution of cycloalkyl and 4-6 membered heterocyclic groups;

[0161] Preferably, R 3 Selected from methyl, trifluoromethyl, ethyl, amino, methylamino, trifluoromethoxy, The wavy line represents the point of attachment of the group to the rest of the molecule.

[0162] In certain embodiments, in the compound of Formula V, V-1, V-1a, V-1b or V-1c,

[0163] R 3 Selected from C 1-6 Haloalkyl and C 1-6 haloalkoxy;

[0164] Preferably, R 3 Selected from trifluoromethyl and trifluoromethoxy.

[0165] In certain embodiments, the compound of formula I provided herein is a compound of formula VI:

[0166] where X 1 、X 2 、X 3 、R1 、R 3 , m is as defined above for the compound of formula I; and

[0167] p is 0 or 1.

[0168] In certain embodiments, the compound of formula VI provided herein is a compound of formula VI-1:

[0169] where R 3 As defined above for compounds of formula I; and

[0170] p is 0 or 1.

[0171] In certain embodiments, the compound of formula VI-1 provided herein is a compound of formula VI-1a:

[0172] where R 3 As defined above for compounds of formula I; and

[0173] p is 0 or 1.

[0174] In certain embodiments, the compound of formula VI-1 provided herein is a compound of formula VI-1b:

[0175] where R 3 As defined above for compounds of formula I; and

[0176] p is 0 or 1.

[0177] In certain embodiments, in the compound of Formula VI, VI-1, VI-1a or VI-1b,

[0178] R 3 Selected from halogen, -CN, -NR 4 R 5 、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, 3-8 membered heterocyclic group, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the alkyl, heteroalkyl, heterocyclyl, aryl or heteroaryl are each optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 3-8Substitution of cycloalkoxy and 3-8 membered heterocyclic groups;

[0179] Preferably, R 3 Selected from halogen, -NR 4 R 5 、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 2-6 heteroalkyl, 4-7 membered heterocyclyl, phenyl and 5-6 membered heteroaryl, wherein each of the alkyl, heteroalkyl, heterocyclyl, phenyl or heteroaryl is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Substitution of cycloalkyl and 4-6 membered heterocyclic groups;

[0180] Preferably, R 3 Selected from-NR 4 R 5 、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, 4-7 membered heterocyclic group and 5-6 membered heteroaryl, the alkyl, heterocyclic group, heteroaryl are optionally substituted with one or more -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Substitution of cycloalkyl and 4-6 membered heterocyclic groups;

[0181] Preferably, R 3 Selected from methyl, trifluoromethyl, ethyl, amino, methylamino, trifluoromethoxy, The wavy line represents the point of attachment of the group to the rest of the molecule.

[0182] In certain embodiments, in the compound of Formula VI, VI-1, VI-1a or VI-1b,

[0183] R 3 is selected from 4-7 membered heterocyclyl and 5-6 membered heteroaryl, wherein the heterocyclyl or aryl is each optionally selected from C 1-6 Alkyl and C 3-6 Substitution of cycloalkyl groups;

[0184] Preferably, R 3 Selected from The wavy line represents the point of attachment of the group to the rest of the molecule.

[0185] In certain embodiments, the compound of formula I provided herein is a compound of formula III-1a:

[0186] where X 1 、X 2 、X 3 、R 1 、R 3 , m are as defined above for the compound of formula I.

[0187] In certain embodiments, the compound of formula I provided herein is a compound of formula V-1a-1:

[0188] where R 3 As defined above for compounds of formula I.

[0189] In certain embodiments, the compound of formula I provided herein is a compound of formula V-1a-2:

[0190] where R 3 As defined above for compounds of formula I.

[0191] In certain embodiments, the compound of formula I provided herein is a compound of formula V-1b-1:

[0192] where R 3 As defined above for compounds of formula I.

[0193] In certain embodiments, the compound of formula I provided herein is a compound of formula V-1b-2:

[0194] where R 3 As defined above for compounds of formula I.

[0195] In certain embodiments, the compound of formula I provided herein is a compound of formula V-1c-1:

[0196] where R 3 As defined above for compounds of formula I.

[0197] In certain embodiments, the compound of formula I provided herein is a compound of formula V-1c-2:

[0198] where R 3As defined above for compounds of formula I.

[0199] In certain embodiments, the compound of formula I provided by the present invention is a compound of formula VI-1a-1:

[0200] where R 3 As defined above for compounds of formula I.

[0201] In certain embodiments, the compound of formula I provided by the present invention is a compound of formula VI-1a-2:

[0202] where R 3 As defined above for compounds of formula I.

[0203] In certain embodiments, the compound of formula I provided by the present invention is a compound of formula VI-1a-3:

[0204] where R 3 As defined above for compounds of formula I.

[0205] The present invention covers any combination of the above embodiments.

[0206] In some embodiments, compounds of the present invention include, but are not limited to:

[0207] Preparation method

[0208] The compounds of the present invention can be prepared by any method known in the art. Reagents and starting materials are readily available to those of ordinary skill in the art. Individual isomers, enantiomers, and diastereomers can be separated or split at any convenient point in the synthesis by methods such as selective crystallization techniques or chiral chromatography (See for example, J. Jacques, et al., "Enantiomers, Racemates, and Resolutions", John Wiley and Sons, Inc., 1981, and E.L. Elel and SH. Wilen).

[0209] In certain embodiments, the present invention provides a method for preparing a compound of Formula I, comprising the following steps:

[0210] Step 1: Compound IA-1 and acyl chloride Compound IA-2 is generated through condensation reaction;

[0211] Step 2: Compound IA-2 is hydrolyzed to generate compound IA-3;

[0212] Step 3: Compound IA-3 and compound IA-4 undergo condensation reaction to produce compound IA-5;

[0213] Step 4: The condensation product IA-5 is deprotected to generate compound I;

[0214] in

[0215] PG is an amino protecting group;

[0216] X 1 、X 2 、X 3 , Ring A, R 1 、R 2 , m, n are as defined above, R 6 and R 7 For H.

[0217] In some embodiments of the present invention, the amino protecting group is p-methoxybenzyl or 2,4-dimethoxybenzyl.

[0218] first step

[0219] In some embodiments of the present invention, the condensation reaction in the first step is carried out in the presence of a base, preferably in the presence of a base such as triethylamine or diisopropylethylamine.

[0220] In some embodiments of the present invention, the first step reaction is carried out in a solvent such as THF.

[0221] Step 2

[0222] In some embodiments of the present invention, the hydrolysis reaction in the second step is carried out in an aqueous base solution, preferably in an aqueous base solution such as LiOH, NaOH, etc.

[0223] In some embodiments of the present invention, the hydrolysis reaction in the second step is carried out in a solvent such as THF, methanol or ethanol.

[0224] Step 3

[0225] In some embodiments of the present invention, the condensation reaction in the third step is carried out in the presence of a condensing agent, preferably in the presence of one or more condensing agents such as HATU, PyBOP, T3P, EDCI, PyBrOP, etc.

[0226] In some embodiments of the present invention, the condensation reaction in the third step is carried out in the presence of a base, preferably in the presence of a base such as triethylamine or diisopropylethylamine.

[0227] In some embodiments of the present invention, the condensation reaction in the third step is carried out in a solvent such as DMF or NMP.

[0228] Step 4

[0229] In some embodiments of the present invention, the deprotection reaction in the fourth step is preferably carried out under the action of an acid, preferably an acid such as aqueous hydrochloric acid solution, hydrochloric acid-ethyl acetate solution, hydrochloric acid-1,4-dioxane solution or trifluoroacetic acid, or oxidative deprotection is performed, preferably under the action of an oxidant such as DDQ.

[0230] Those skilled in the art will appreciate that, depending on the desired product structure, one or more steps in the above-described preparation method may be omitted, and the order of the reaction steps may be appropriately adjusted, and protection / deprotection reaction steps may be added or omitted as needed.

[0231] Pharmaceutical compositions, preparations and methods of treatment

[0232] In some embodiments, the present invention provides a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof and one or more pharmaceutically acceptable carriers.

[0233] In some embodiments, the present invention provides a pharmaceutical preparation comprising a preventive or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof and one or more pharmaceutically acceptable carriers. The pharmaceutical preparation is preferably a solid preparation, a semisolid preparation, a liquid preparation or a gaseous preparation.

[0234] In some embodiments, the pharmaceutical composition or pharmaceutical formulation may further comprise one or more additional therapeutic agents.

[0235] In some embodiments, the pharmaceutical composition or pharmaceutical formulation is preferably administered orally, intravenously, intraarterially, subcutaneously, intraperitoneally, intramuscularly, or transdermally.

[0236] In some embodiments, the present invention provides use of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, or a pharmaceutical composition of the present invention, or a pharmaceutical formulation of the present invention in the preparation of a medicament for preventing or treating a disease or condition associated with PRMT5 activity.

[0237] In some embodiments, the present invention provides use of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, or a pharmaceutical composition of the present invention, or a pharmaceutical formulation of the present invention in the preparation of a medicament for modulating (e.g., reducing or inhibiting) PRMT5 activity.

[0238] In some embodiments, the present invention provides a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, or a pharmaceutical composition of the present invention, or a pharmaceutical formulation of the present invention, for use in preventing or treating a disease or condition associated with PRMT5 activity.

[0239] In some embodiments, the present invention provides a method for preventing or treating a disease or condition associated with PRMT5 activity, comprising administering to a subject in need thereof an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, or a pharmaceutical composition of the present invention, or a pharmaceutical formulation of the present invention.

[0240] In some embodiments, the disease or condition associated with PRMT5 activity is preferably a cancer or tumor with MTAP deficiency.

[0241] In some embodiments, the cancer or tumor is preferably esophageal cancer, lung cancer, pancreatic cancer, glioblastoma, bile duct cancer, bladder cancer, breast cancer, ovarian cancer, hepatocellular carcinoma, prostate cancer, melanoma, gastric cancer, colon cancer, leukemia (particularly chronic B-lymphocytic leukemia (B-CLL)), lymphoma, etc.

[0242] In the present invention, "pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient or vehicle that is administered together with the therapeutic agent and is suitable for contact with the tissues of humans and / or other animals without excessive toxicity, irritation, allergic response or other problems or complications corresponding to a reasonable benefit / risk ratio within the scope of reasonable medical judgment.

[0243] Pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, sterile liquids. Examples of suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (1990).

[0244] The pharmaceutical compositions of the present invention can act systemically and / or locally. For this purpose, they can be administered by any suitable route.

[0245] For these administration routes, the pharmaceutical composition of the present invention can be administered in suitable dosage forms.

[0246] As used herein, the term "effective amount" refers to that amount of a compound which, when administered, will relieve to some extent one or more of the symptoms of the condition being treated.

[0247] The dosage regimen can be adjusted to provide the optimal desired response. For example, a single bolus can be administered, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the urgency of the therapeutic situation. It is to be noted that dosage values ​​can vary with the type and severity of the condition to be alleviated and can include single or multiple doses. It is to be further understood that for any particular individual, the specific dosage regimen should be adjusted over time according to the individual's needs and the professional judgment of the person administering or supervising the administration of the composition.

[0248] The amount of the compound of the present invention administered will depend on the severity of the individual, disease or the patient's condition, the speed of administration, the disposal of the compound and the judgment of the prescribing physician for treatment. Generally speaking, effective dose is about 0.0001 to about 50 mg per kg body weight per day. In some cases, the dosage level not higher than the lower limit of the aforementioned range can be enough, and in other cases, still can adopt larger doses when not causing any harmful side effects, condition is first divided into several smaller doses to be administered throughout the day.

[0249] The compound of the present invention may be contained in a pharmaceutical composition or formulation in an amount ranging from about 0.01 mg to about 1000 mg.

[0250] As used herein, unless otherwise indicated, the term "treating" means reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition.

[0251] As used herein, "subject" includes humans and non-human animals. Exemplary human subjects include human subjects suffering from diseases (e.g., the diseases described herein) (referred to as patients) or normal individuals. "Non-human animals" herein include all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

[0252] In some embodiments, the pharmaceutical compositions or pharmaceutical formulations of the present invention may further comprise one or more additional therapeutic or prophylactic agents (e.g., other drugs used to treat cancer or tumor diseases). In some embodiments, the therapeutic methods of the present invention may further comprise administering one or more additional therapeutic or prophylactic agents (e.g., other drugs used to treat cancer or tumor diseases). DETAILED DESCRIPTION

[0253] Example

[0254] The present invention is further described below with reference to examples, but these examples are not intended to limit the scope of the present invention.

[0255] The abbreviations used in this document have the following meanings:

[0256] The compounds of the present invention are separated and purified by preparative TLC, silica gel column chromatography, Prep-HPLC and / or flash column chromatography (Flash column chromatography). 1 The reaction was confirmed by H NMR and / or MS. Reaction monitoring was performed by TLC or LC-MS.

[0257] 1 H NMR spectroscopy was performed using a Bruker superconducting nuclear magnetic resonance spectrometer (model AVACE III HD 400 MHz).

[0258] LC / MS uses Aglient 1260 Infinity / Aglient 6120 Quadrupole.

[0259] TLC used silica gel GF 254 as the stationary phase.

[0260] Column chromatography generally uses 200-300 mesh silica gel (Qingdao Ocean) as the stationary phase.

[0261] Flash column chromatography was performed using a Biotage flash column chromatograph.

[0262] Prep-HPLC used Agilent 1260 and Waters 2489.

[0263] Microwave reactions were performed using a BiotageInitiator microwave reactor.

[0264] In the following examples, unless otherwise specified, the reaction temperature is room temperature (15-30°C).

[0265] The reagents used in this application were purchased from Acros Organics, Aldrich Chemical Company, or Teber Chemical Company.

[0266] Synthesis Example:

[0267] Intermediate Int A: 2-((4-((tert-Butyloxycarbonyl)amino)-1,3-dihydrofuro[3,4-c]pyridin-7-yl)amino)-2-oxoacetic acid

[0268] Step 1: Synthesis of dimethyl pyridine-3,4-dicarboxylate (Compound Int A-2)

[0269] Int A-1 (30 g, 179.51 mmol) and MeOH (300 mL) were added to a reaction flask, followed by the slow addition of SOCl2 (42.71 g, 359.03 mmol, 26.04 mL). The temperature was raised to 80°C and the reaction mixture was allowed to react for 3 hours. The reaction mixture was spin-dried to dryness, and additional SOCl2 (42.71 g, 359.03 mmol, 26.04 mL) and MeOH (300 mL) were added to the reaction flask and the reaction mixture was allowed to react at 80°C for 3 hours. After the reaction was completed, the reaction mixture was spin-dried to dryness, dissolved in ethyl acetate (500 mL), and washed twice with aqueous sodium bicarbonate solution. The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound Int A-2 (32.46 g). MS (ESI, m / z): 196.1 [M+H] + .

[0270] Step 2: Synthesis of 3,4-bis(methoxycarbonyl)pyridine-1-oxide (Compound Int A-3)

[0271] Int A-2 (4.4 g, 22.54 mmol) was dissolved in DCM (50 mL), and mCPBA (7.29 g, 33.82 mmol) was added portionwise. The mixture was stirred at 25°C overnight. After completion of the reaction, sodium thiosulfate aqueous solution was added to the reaction solution for quenching. The mixture was stirred for 2 hours and extracted with dichloromethane (200 ml x 3). The organic phases were combined, dried over sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain compound Int A-3 (4.7 g). MS (ESI, m / z): 212.1 [M+H] + .

[0272] Step 3: Synthesis of methyl 2-chloropyridine-3,4-dicarboxylate (Compound Int A-4)

[0273] Int A-3 (4.7 g, 22.26 mmol) and POCl3 (20 mL) were added to a reaction flask and reacted at 100°C overnight. After the reaction was completed, the reaction solution was evaporated under reduced pressure to dryness and slowly poured into water. The pH was adjusted to 7-8 with sodium bicarbonate. Extraction was performed with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (EA:PE = 1:10) to obtain compound Int A-4 (2.2 g). MS (ESI, m / z): 230.0 [M+H] + .

[0274] Step 4: Synthesis of (2-chloropyridine-3,4-diyl)dimethanol (Compound Int A-5)

[0275] Int A-4 (1.5 g, 6.53 mmol) was dissolved in EtOH (20 mL), and NaBH4 (1.48 g, 39.20 mmol) was added portionwise. The reaction was allowed to proceed overnight at 25°C. After completion of the reaction, the pH was adjusted to ~5 with formic acid, and the solvent was evaporated under reduced pressure. The product was purified by silica gel column chromatography (PE:EA = 25:75) to obtain compound Int A-5 (800 mg). MS (ESI, m / z): 174.1 [M+H] + .

[0276] Step 5: Synthesis of 4-chloro-1,3-dihydrofuro[3,4-c]pyridine (Compound Int A-6)

[0277] Int A-5 (1.15 g, 6.62 mmol) was dissolved in DCM (25 mL), and MnO2 (1.15 g, 13.25 mmol) was added portionwise. Et3SiH (5 mL) and trifluoroacetic acid (9 mL) were added dropwise, and the mixture was allowed to react overnight at 25°C. After completion of the reaction, the solvent was evaporated under reduced pressure, and the crude product was purified by silica gel column chromatography (EA:PE = 20:80) to obtain compound Int A-6 (200 mg). MS (ESI, m / z): 156.1 [M+H] + .

[0278] Step 6: Synthesis of N-(2,4-dimethoxybenzyl)-1,3-dihydrofuro[3,4-c]pyridin-4-amine (Compound Int A-8)

[0279] Int A-6 (200 mg, 1.29 mmol), Int A-7 (430 mg, 2.57 mmol), toluene (5 mL), tBuONa (494 mg, 5.14 mmol), BINAP (80 mg, 128.55 μmol), and Pd2(dba)3 (59 mg, 64.28 μmol) were added sequentially to a reaction flask. The atmosphere was purged with nitrogen three times and the reaction was allowed to proceed overnight at 100°C. After completion of the reaction, the reaction solution was spin-dried, diluted with water, and extracted three times with ethyl acetate. The organic phases were combined and dried over sodium sulfate. The solvent was removed by distillation under reduced pressure, and the product was purified by silica gel column chromatography (PE:EA = 75:25) to obtain compound Int A-8 (230 mg). MS (ESI, m / z): 287.1 [M+H] + .

[0280] Step 7: Synthesis of 1,3-dihydrofuro[3,4-c]pyridin-4-amine (Compound Int A-9)

[0281] Int A-8 (230 mg, 803.28 μmol) was dissolved in TFA (10 mL) and reacted at 25°C for 1 hour. After completion of the reaction, the reaction solution was spin-dried and diluted with saturated aqueous NaHCO3 solution. The mixture was extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to obtain compound Int A-9 (96 mg).

[0282] Step 8: Synthesis of 7-bromo-1,3-dihydrofuro[3,4-c]pyridin-4-amine (Compound Int A-10)

[0283] Int A-9 (96 mg, 705.10 μmol) and MeCN (3 mL) were added to a reaction flask, followed by the addition of NBS (138 mg, 775.61 μmol) in portions. The mixture was allowed to react at 25°C for 2 hours. After the reaction, the solvent was removed by distillation under reduced pressure, and the product was purified by silica gel column chromatography (PE:EA = 70:30) to obtain compound Int A-10 (110 mg). MS (ESI, m / z): 215.0 [M+H] + .

[0284] Step 9: Synthesis of 7-bromo-N,N-bis(tert-butyloxycarbonyl)-1,3-dihydrofuro[3,4-c]pyridin-4-amine (Compound Int A-11)

[0285] Int A-10 (100 mg, 465.01 μmol), DCM (5 mL), TEA (141 mg, 1.40 mmol), and DMAP (11.36 mg, 93.00 μmol) were added to a reaction flask. Boc2O (254 mg, 1.16 mmol) was added portionwise and reacted at 25°C for 2 hours. After the reaction was completed, the reaction solution was diluted with water and extracted with DCM (30 ml x 3). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (EA:PE = 10:90) to obtain compound Int A-11 (83 mg). MS (ESI, m / z): 415.1 [M+H] + .

[0286] Step 10: Synthesis of tert-butyl (7-((diphenylmethylene)amino)-1,3-dihydrofuro[3,4-c]pyridin-4-yl)carbamate (Compound Int A-13)

[0287] Int A-11 (90 mg, 216.72 μmol), Int A-12 (79 mg, 433.44 μmol), tBuONa (63 mg, 650.17 μmol), BINAP (14 mg, 21.67 μmol), Pd2(dba)3 (10 mg, 10.84 μmol), and 1,4-dioxane (5 mL) were added sequentially to a reaction flask. The atmosphere was purged with nitrogen three times and the reaction was allowed to proceed at 80°C overnight. After completion of the reaction, the solvent was evaporated under reduced pressure, and the mixture was purified by silica gel column chromatography (EA:PE = 15:85) to obtain compound Int A-13 (50 mg). MS (ESI, m / z): 416.2 [M+H] + .

[0288] Step 11: Synthesis of tert-butyl (7-amino-1,3-dihydrofuro[3,4-c]pyridin-4-yl)carbamate (Compound Int A-14)

[0289] Int A-13 (50 mg, 120.34 μmol), Pd / C (14.62 mg, 120.34 μmol), MeOH (3 mL), and THF (1 mL) were added to the reaction flask. The mixture was purged with a hydrogen balloon three times and allowed to react at 60°C for 5 hours. After completion of the reaction, the reaction solution was filtered through a filter and diluted with water. The mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to yield compound Int A-14 (30 mg). MS (ESI, m / z): 252.2 [M+H] + .

[0290] Step 12: Synthesis of ethyl 2-((4-((tert-Butyloxycarbonyl)amino)-1,3-dihydrofuro[3,4-c]pyridin-7-yl)amino)-2-oxoacetate (Compound Int A-16)

[0291] To the reaction flask, Int A-14 (30 mg, 119.39 μmol), DIPEA (38.57 mg, 298.47 μmol), and THF (3 mL) were added, followed by the slow addition of Int A-15 (17 mg, 119.39 μmol). The mixture was incubated at 25°C for 1 hour under nitrogen. After completion of the reaction, the mixture was diluted with water and extracted with ethyl acetate (30 ml x 3). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (EA:PE = 35:65) to obtain compound Int A-16 (23 mg). MS (ESI, m / z): 352.2 [M+H] + .

[0292] Step 13: Synthesis of 2-((4-((tert-Butyloxycarbonyl)amino)-1,3-dihydrofuro[3,4-c]pyridin-7-yl)amino)-2-oxoacetic acid (Compound Int A)

[0293] Int A-16 (23 mg, 65.46 μmol) and THF (1 mL) were added to a reaction flask. After dissolution, a solution of LiOH·H₂O (6 mg, 130.92 μmol) in H₂O (0.5 mL) was added. Under nitrogen, the mixture was reacted at 25°C for 2 hours. After completion of the reaction, the pH was adjusted to 6-7 with 2M hydrochloric acid, and the reaction solution was directly spin-dried to obtain compound Int A (20 mg). MS (ESI, m / z): 324.1 [M+H] + .

[0294] Intermediate Int B: 2-((7-((2,4-dimethoxybenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)amino)-2-oxoacetic acid

[0295] Step 1: Synthesis of 4-bromo-7-chloro-1-methyl-1H-pyrazolo[3,4-c]pyridine (Compound Int B-2)

[0296] Int B-1 (5 g, 21.51 mmol) was dissolved in DMF (30 mL) and NaH (1.72 g, 43.02 mmol) was slowly added under ice. The mixture was allowed to react for 30 minutes after addition, and then MeI (3.66 g, 25.81 mmol) was added. Stirring was continued at 25°C for 2 hours after addition. After the reaction was completed, water was added to the reaction solution to quench it. The mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 70:30) to obtain compound Int B-2 (3.0 g). MS (ESI, m / z): 245.9 [M+H] + .

[0297] Step 2: Synthesis of 4-bromo-N-(2,4-dimethoxybenzyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-7-amine (Compound Int B-3)

[0298] Int B-2 (3.0 g, 12.17 mmol), Int A-7 (3.05 g, 18.26 mmol) and DIPEA (2.36 g, 18.26 mmol, 3.2 mL) were dissolved in NMP (10 mL), replaced with nitrogen, and stirred at 140°C for 5 hours. After the reaction was completed, water was added to the reaction solution to quench it, and it was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with water and saturated brine in sequence, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Purification by silica gel column chromatography (PE:EA = 70:30) gave compound Int B-3 (4.0 g). MS (ESI, m / z): 377.0 [M+H] + .

[0299] Step 3: Synthesis of N-(2,4-dimethoxybenzyl)-4-((diphenylmethylene)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-7-amine (Compound Int B-4)

[0300] Int B-3 (4.0 g, 10.60 mmol), Int A-12 (3.84 g, 21.21 mmol), tBuONa (3.06 g, 31.81 mmol), BINAP (660 mg, 1.06 mmol), and Pd2(dba)3 (485 mg, 0.53 mmol) were dissolved in toluene (10 mL). After replacing the atmosphere with nitrogen, the reaction temperature was raised to 80°C and stirred for 16 hours. After completion of the reaction, the mixture was concentrated to dryness under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (EA:PE = 90:10) to obtain compound Int B-4 (4.68 g). MS (ESI, m / z): 478.2 [M+H] + .

[0301] Step 4: N 7 Synthesis of -(2,4-dimethoxybenzyl)-1-methyl-1H-pyrazolo[3,4-c]pyridine-4,7-diamine (Compound Int B-5)

[0302] Int B-4 (4.68 g, 9.80 mmol) was dissolved in MeOH (30 mL), followed by the addition of a 4N HCl solution in 1,4-dioxane (3 mL). After nitrogen displacement, the reaction was stirred at 25°C for 1 hour. After completion of the reaction, the mixture was concentrated to dryness under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (DCM:MeOH = 90:10) to obtain compound Int B-5 (3 g). MS (ESI, m / z): 314.1 [M+H] + .

[0303] Step 5: Synthesis of ethyl 2-((7-((2,4-dimethoxybenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)amino)-2-oxoacetate (Compound Int B-6)

[0304] Int B-5 (3 g, 9.57 mmol) and DIPEA (3.71 g, 28.72 mmol) were dissolved in THF (25 mL), and ethyl oxalyl chloride (1.57 g, 11.49 mmol) was slowly added. The mixture was stirred at 25°C for 0.5 hr. After the reaction was completed, water was added to quench the reaction solution and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (EA:PE = 50:50) to obtain compound Int B-6 (1.38 g). MS (ESI, m / z): 414.1 [M+H] + .

[0305] Step 6: Synthesis of 2-((7-((2,4-dimethoxybenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)amino)-2-oxoacetic acid (Compound Int B)

[0306] Int B-6 (1.38 g, 3.34 mmol) and LiOH·H2O (280 mg, 6.68 mmol) were added to THF (15 mL) and H2O (3 mL). The mixture was stirred at 25°C for 2 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure to remove the solvent, yielding compound Int B (1 g). MS (ESI, m / z): 386.1 [M+H] + .

[0307] Intermediate Int C: 2-((4-((4-methoxybenzyl)amino)-1-methyl-1H-pyrazolo[4,3-c]pyridin-7-yl)amino)-2-oxoacetic acid

[0308] Step 1: Synthesis of 2-amino-5-bromo-4-chloronicotinaldehyde (Compound Int C-2)

[0309] Dissolve Int C-1 (1 g, 6.39 mmol) in anhydrous DCE (20 mL) and add NBS (1.25 g, 7.03 mmol). After N protection, heat to 60°C and react for 2 hours. After completion of the reaction, concentrate under reduced pressure to remove the solvent, dissolve in water, and extract with ethyl acetate. The organic layer is dried and concentrated to yield the intermediate Int C-2 (1.4 g).

[0310] Step 2: Synthesis of 7-bromo-1-methyl-1H-pyrazolo[4,3-c]pyridin-4-amine (Compound Int C-4)

[0311] Int C-2 (1.5 g, 6.37 mmol) and methylhydrazine sulfate (Int C-3, 1.38 g, 9.56 mmol) were dissolved in EtOH (30 mL). DIPEA (4.12 g, 31.85 mmol) was added. The mixture was heated to 80°C under N2 protection and allowed to react for 24 hours. After completion of the reaction, the mixture was concentrated under reduced pressure. The crude product was isolated and purified by silica gel column chromatography (DCM:MeOH = 80:20) to obtain the intermediate Int C-4 (320 mg). MS (ESI, m / z): 226.9 [M+H] + .

[0312] Step 3: Synthesis of 7-bromo-N-(4-methoxybenzyl)-1-methyl-1H-pyrazolo[4,3-c]pyridin-4-amine (Int C-5)

[0313] Int C-4 (280 mg, 1.23 mmol) and DIPEA (318.74 mg, 2.47 mmol) were dissolved in anhydrous NMP (10 mL), and PMBCl (289.68 mg, 1.85 mmol) was added. After addition, the mixture was heated to 140°C under N2 protection and allowed to react for 5 hours. After completion, the reaction was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The crude product was separated and purified by normal phase flash chromatography (PE:EA = 92:8) to obtain intermediate Int C-5 (150 mg). MS (ESI, m / z): 347.0 [M+H] + .

[0314] Step 4: Synthesis of 7-((diphenylmethylene)amino)-N-(4-methoxybenzyl)-1-methyl-1H-pyrazolo[4,3-c]pyridin-4-amine (Compound Int C-6)

[0315] Int C-5 (200 mg, 576.02 μmol), Int E-5 (208.79 mg, 1.15 mmol), tBuONa (166.07 mg, 1.73 mmol), BINAP (35.87 mg, 57.60 μmol), and Pd2(dba)3 (26.37 mg, 28.80 μmol) were dissolved in toluene (10 mL). Under nitrogen protection, the temperature was raised to 80°C and the reaction was allowed to react for 16 hours. After completion of the reaction, the toluene was removed by concentration under reduced pressure. The crude product was isolated and purified by normal phase flash chromatography (PE:EA = 79:21) to obtain the intermediate Int C-6 (80 mg). MS (ESI, m / z): 448.2 [M+H] + .

[0316] Step 5: N 4 Synthesis of -(4-methoxybenzyl)-1-methyl-1H-pyrazolo[4,3-c]pyridine-4,7-diamine (Compound Int C-7)

[0317] Int C-6 (80 mg, 178.76 μmol) was dissolved in methanol (3 mL), and a 4N HCl solution in 1,4-dioxane (0.3 mL) was added dropwise. The reaction system was allowed to react at 25°C for 1 hour. After completion of the reaction, the solvent was removed by concentration under reduced pressure. An appropriate amount of methanol was added for dissolution, and a few drops of TEA were added to adjust the pH to 7-8. The reaction was then concentrated under reduced pressure. The crude product was isolated and purified by normal phase flash chromatography (DCM:MeOH = 93:7) to obtain the intermediate Int C-7 (50 mg). MS (ESI, m / z): 284.1 [M+H] + .

[0318] Step 6: Synthesis of ethyl 2-((4-((4-methoxybenzyl)amino)-1-methyl-1H-pyrazolo[4,3-c]pyridin-7-yl)amino)-2-oxoacetate (Compound Int C-8)

[0319] Int C-7 (90 mg, 317.65 μmol) and DIPEA (41.05 mg, 317.65 μmol) were dissolved in THF (10 mL). Ethyl oxalyl chloride (52.04 mg, 381.18 μmol) was slowly added. The reaction system was allowed to react at 25°C for 1 hour. After completion of the reaction, the solvent was removed by concentration under reduced pressure. The crude product was isolated and purified by normal phase flash chromatography (DCM:MeOH = 90:10) to obtain the intermediate Int C-8 (110 mg). MS (ESI, m / z): 384.2 [M+H] + .

[0320] Step 7: Synthesis of 2-((4-((4-methoxybenzyl)amino)-1-methyl-1H-pyrazolo[4,3-c]pyridin-7-yl)amino)-2-oxoacetic acid (Compound Int C)

[0321] Int C-8 (120 mg, 312.99 μmol) was dissolved in THF (5 mL) and H₂O (1 mL). NaOH (25.04 mg, 625.98 μmol) was added. After addition, the reaction system was heated to 25°C and allowed to react for 2 hours. After completion of the reaction, the solvent was removed by concentration under reduced pressure to obtain the intermediate Int C (115 mg). MS (ESI, m / z): 356.1 [M+H] + .

[0322] Intermediate Int D: 2-(5-methylpiperidin-2-yl)-5-(trifluoromethyl)pyridine

[0323] Step 1: Synthesis of 5-methylpiperidin-2-one (Compound Int D-2)

[0324] Int D-1 (5 g, 45.82 mmol) was dissolved in MeOH (40 mL), and then Pd(OH)2 / C (6.43 g, 4.58 mmol, 10% purity) was added. The hydrogen atmosphere was replaced three times. The autoclave was heated to 80°C and stirred for 12 hours. After the reaction, the mixture was filtered and the filtrate was concentrated to obtain compound Int D-2 (5 g). MS (ESI, m / z): 114.2 [M+H] + .

[0325] Step 2: Synthesis of tert-butyl 5-methyl-2-oxopiperidine-1-carboxylate (Compound Int D-3)

[0326] Int D-2 (1 g, 8.84 mmol) and DMAP (107.97 mg, 883.73 μmol) were added to THF (10 mL), followed by the slow addition of (Boc)2O (2.12 g, 9.72 mmol). The mixture was stirred at 25°C for 16 hours. After completion of the reaction, the mixture was quenched with ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel flash column chromatography (PE:EA = 9:1) to afford compound Int D-3 (1.72 g).

[0327] Step 3: Synthesis of tert-butyl 3-methyl-6-(((trifluoromethyl)sulfonyl)oxy)-3,4-dihydropyridine-1(2H)-carboxylate (Compound Int D-5)

[0328] Int D-3 (5 g, 23.44 mmol) was dissolved in THF (50 mL) and cooled to -78°C. LiHMDS (35.17 mmol, 1 M THF solution, 35.17 mL) was then slowly added dropwise. After the addition was complete, the mixture was stirred at -78°C for 0.5 hr. A solution of Int D-4 (10.05 g, 28.13 mmol) in THF (20 mL) was then added. After the addition was complete, the temperature was naturally raised to 25°C and the reaction was allowed to proceed for 16 hr. After completion of the reaction, saturated aqueous ammonium chloride was added to quench the reaction under ice-cooling. The mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel flash column chromatography (PE:EA = 9:1) to afford compound Int D-5 (7.5 g). MS (ESI, m / z): 363.1 [M+18]. + .

[0329] Step 4: Synthesis of tert-butyl 3-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate (Compound Int D-7)

[0330] Int D-5 (7.5 g, 21.72 mmol), Int D-6 (8.27 g, 32.58 mmol), PPh3 (341.78 mg, 1.30 mmol), Pd(PPh3)4 (752.90 mg, 651.54 μmol), and K2CO3 (7.50 g, 54.30 mmol) were weighed into a two-necked flask. 1,4-dioxane (50 mL) was then added. After nitrogen displacement, the reaction mixture was heated to 90°C and stirred for 20 hours. The solvent was removed by concentration, and the mixture was then extracted three times with dichloromethane. The combined organic phases were concentrated and purified by reverse-phase column chromatography (H2O [0.05% NH4HCO3]:ACN = 10:90). After concentration, the mixture was extracted with ethyl acetate, dried, filtered, and concentrated to afford compound Int D-7 (1.55 g). MS (ESI, m / z): 324.2 [M+H] + .

[0331] Step 5: Synthesis of tert-butyl 5-methyl-5'-(trifluoromethyl)-5,6-dihydro-[2,2'-bipyridine]-1(4H)-carboxylate (Compound Int D-9)

[0332] Int D-8 (2.17 g, 9.59 mmol), Int D-7 (1.55 g, 4.80 mmol), and Pd(dppf)Cl2·DCM (391.60 mg, 479.53 μmol) were dissolved in 1,4-dioxane (50 mL). A solution of Na2CO3 (1.02 g, 9.59 mmol) in H2O (10 mL) was then added. After nitrogen was replaced, the reaction system was heated to 90°C for 2 hours. The reaction mixture was cooled to room temperature, concentrated, and extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to afford compound Int D-9 (1.6 g), which was used directly in the next reaction. MS (ESI, m / z): 343.1 [M+H] + .

[0333] Step 6: Synthesis of 5-methyl-5'-(trifluoromethyl)-3,4,5,6-tetrahydro-2,2'-bipyridine (Compound Int D-10)

[0334] Int D-9 (1.6 g, 4.67 mmol) was added to a 4N HCl solution in 1,4-dioxane (10 mL) and reacted at 25°C for 2 hours. The mixture was concentrated to afford compound Int E-3 (1 g), which was used directly in the next reaction. MS (ESI, m / z): 243.1 [M+H] + .

[0335] Step 7: Synthesis of 2-(5-methylpiperidin-2-yl)-5-(trifluoromethyl)pyridine (Compound Int D)

[0336] Int D-10 (1 g, 4.13 mmol) was added to THF (10 mL). After nitrogen displacement, NaBH4 (312.36 mg, 8.26 mmol) was added and the reaction was stirred at 25°C for 0.5 hr. After completion, saturated ammonium chloride was added dropwise to quench the reaction. The mixture was concentrated and purified by reverse-phase column chromatography (H2O [0.05% HCO2H]:ACN = 70:30). After lyophilization, the target compound Int D (500 mg) was obtained. MS (ESI, m / z): 245.1 [M+H] + .

[0337] Intermediate Int E: 5-(5-methylpiperidin-2-yl)benzo[d]thiazole

[0338] Step 1: Synthesis of 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazole (Compound Int E-2)

[0339] Int E-1 (500 mg, 2.34 mmol), Int D-6 (771.01 mg, 3.04 mmol), Pd(dppf)Cl2·DCM (190.73 mg, 233.56 μmol), and KOAc (687.63 mg, 7.01 mmol) were added to 1,4-dioxane (30 mL). After nitrogen displacement, the reaction system was heated to 80°C for 2 hours. The reaction solution was directly separated and purified by silica gel column chromatography (PE:EA = 98:2) and concentrated to obtain compound Int E-2 (580 mg). MS (ESI, m / z): 262.1 [M+H] + .

[0340] Step 2: Synthesis of tert-butyl 6-(benzo[d]thiazol-5-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylate (Compound Int E-3)

[0341] Int E-2 (100 mg, 382.93 μmol), Int D-5 (132.24 mg, 382.93 μmol), Pd(dppf)Cl2·DCM (31.27 mg, 38.29 μmol), and Na2CO3 (81.17 mg, 765.85 μmol) were added to a mixture of 1,4-dioxane (10 mL) and H2O (1 mL). The reaction system was heated to 90°C for 2 hours. After cooling, the reaction system was directly concentrated to dryness. The crude product was purified by silica gel column chromatography (PE:EA = 20:1) to obtain the target compound Int E-3 (580 mg). MS (ESI, m / z): 331.1 [M+H] + .

[0342] Step 3: Synthesis of 5-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)benzo[d]thiazole (Compound Int E-4)

[0343] Int E-3 (600 mg, 1.82 mmol) was added to a 4N HCl solution in 1,4-dioxane (10 mL) and allowed to react at 25°C for 2 hours. The mixture was concentrated to dryness, methanol and triethylamine were added, and the mixture was concentrated again to dryness. The residue was separated and purified by silica gel column chromatography (DCM:MeOH = 98:2). After concentration, the target compound Int E-4 (385 mg) was obtained. MS (ESI, m / z): 231.1 [M+H] + .

[0344] Step 4: Synthesis of 5-(5-methylpiperidin-2-yl)benzo[d]thiazole (Compound Int E)

[0345] Int E-4 (440 mg, 1.91 mmol) was added to THF (10 mL). After nitrogen was replaced, NaBH4 (144.54 mg, 3.82 mmol) was added and the reaction was stirred at 25°C for 0.5 hr. After completion, saturated ammonium chloride was added dropwise to quench the reaction. The mixture was concentrated to dryness and purified by silica gel column chromatography (DCM:MeOH = 97:3). After concentration, the target compound Int E (400 mg) was obtained. MS (ESI, m / z): 233.1 [M+H] + .

[0346] Intermediates Int F and G: tert-butyl 5-((diphenoxyphosphoryl)oxy)-3-methyl-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound Int F) and tert-butyl 3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound Int G)

[0347] Step 1: Synthesis of 5-methylmorpholin-3-one (Compound Int F-3)

[0348] Under an ice bath, NaH (120 g, 3.00 mol) was slowly added to THF (1.00 L). Then, Int F-1 (100 g, 1.33 mol) was slowly added dropwise to the reaction mixture at 0°C. Stirring was continued for 30 minutes, followed by the dropwise addition of Int F-2 (204 g, 1.66 mol). After the addition was complete, the mixture was heated to 25°C and allowed to react for 16 hours. TLC (PE / EA = 1 / 1, Rf = 0.15) and LCMS confirmed the reaction was complete. After cooling to 0°C, saturated NH4Cl (1.00 L) was slowly added dropwise to quench the reaction. After extraction with ethyl acetate (6.00 L), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to yield compound Int F-3 (158 g). MS (ESI, m / z): 116.3 [M+H] + .

[0349] Step 2: Synthesis of tert-butyl 3-methyl-5-oxomorpholine-4-carboxylate (Compound Int F-4)

[0350] Compound Int F-3 (158 g, 1.37 mol) was dissolved in THF (1.00 L), followed by the addition of DMAP (16.8 g, 137 mmol) and Boc2O (329 g, 1.51 mol). The reaction was stirred at 25°C for 16 hours. LCMS confirmed the reaction was complete. The reaction mixture was quenched with water (200 mL) and concentrated under reduced pressure to remove the THF. The mixture was then extracted with ethyl acetate (900 mL). The combined organic phases were washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was achieved by silica gel column chromatography (PE / EA = 1 / 0 to 6 / 1) to afford compound Int F-4 (55.0 g). MS (ESI, m / z): 159.9 [M-tBu+H] + .

[0351] Step 3: Synthesis of tert-butyl 5-((diphenoxyphosphoryl)oxy)-3-methyl-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound Int F)

[0352] Compound Int F-4 (54.9 g, 255 mmol) was dissolved in THF (800 mL), the atmosphere was replaced with nitrogen, and the temperature was lowered to -70°C. LiHMDS (1 M, 281 mL) was then slowly added dropwise. The reaction mixture was stirred at -70°C for 1 hour before Int F-5 (71.9 g, 268 mmol) was added. After the addition was complete, the reaction mixture was stirred at -60 to -30°C for 3 hours. LCMS confirmed the reaction was complete. The mixture was quenched with water (200 mL) in an ice bath and then extracted with ethyl acetate (300 mL x 3). The organic phases were combined, washed with saturated brine (600 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE / EA = 1 / 0 to 9 / 1) to afford compound Int F (98.9 g). 348.1 [M+H-Boc] + .

[0353] Step 4: Synthesis of tert-butyl 3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound Int G)

[0354] Compound Int F (98.9 g, 221 mmol), B2Pin2 (84.2 g, 332 mmol), KOAc (65.1 g, 663 mmol), and XPhos Pd G3 (18.7 g, 22.1 mmol) were dissolved in 1,4-dioxane (1000 mL). After nitrogen replacement, the reaction temperature was raised to 100°C and stirred for 2 hours. TLC (PE / EtOAc = 5 / 1, R f =0.47) and LCMS analysis showed that the reaction was complete. The reaction mixture was cooled to room temperature, and the solid was removed by filtration. The filter cake was then washed with CH2Cl2 (1.0 L). The organic phases were combined and concentrated by distillation under reduced pressure. The mixture was then separated and purified by silica gel column chromatography (PE / EtOAc = 1 / 0 to 9 / 1) to obtain compound Int G (61.5 g). MS (ESI, m / z): 269.7 [M+H-tBu] + .

[0355] Intermediates Int H: 3-(Benzo[d]thiazol-5-yl)-5-methylmorpholine and (3S,5R)-3-(Benzo[d]thiazol-5-yl)-5-methylmorpholine (Int H peak 1), (3R,5S)-3-(Benzo[d]thiazol-5-yl)-5-methylmorpholine (Int H peak 2)

[0356] Step 1: Synthesis of tert-butyl 5-(benzo[d]thiazol-5-yl)-3-methyl-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound Int H-1)

[0357] Compounds Int F (6.00 g, 13.4 mmol) and Int E-2 (4.20 g, 16.1 mmol) were dissolved in MeCN (80.0 mL) and H2O (20.0 mL). K3PO4 (8.54 g, 40.2 mmol) and Pd(t-Bu3P)2 (685 mg, 1.34 mmol) were then added sequentially under a nitrogen atmosphere. After the addition was complete, the reaction was stirred at 65°C for 12 hours. After LCMS analysis, the reaction was concentrated by vacuum distillation and purified by silica gel column chromatography (PE / EA = 100 / 1 to 5 / 1, R f =0.43) was separated and purified, and concentrated to give compound Int H-1 (2.06 g, 6.20 mmol). MS (ESI, m / z): 333.2 [M+H] + .

[0358] Step 2: Synthesis of 5-(Benzo[d]thiazol-5-yl)-3-methyl-3,4-dihydro-2H-1,4-oxazine (Compound Int H-2)

[0359] Compound Int H-1 (2.06 g, 6.20 mmol) was dissolved in CH2Cl2 (10.0 mL), and TFA (7.68 g, 67.3 mmol, 5.00 mL) was added. The reaction was stirred at 25°C for 0.5 hr. LCMS confirmed the reaction was complete and the mixture was concentrated to yield compound Int H-2 (1.44 g). MS (ESI, m / z): 232.9 [M+H] + .

[0360] Step 3: Synthesis of 3-(Benzo[d]thiazol-5-yl)-5-methylmorpholine (Compound Int H)

[0361] Compound Int H-2 (1.44 g, 6.20 mmol) was dissolved in MeOH (10.0 mL), followed by the addition of NaBH4 (1.23 g, 32.5 mmol) at 0°C. The reaction was stirred at 25°C for 0.5 hr. LCMS confirmed complete conversion of the starting material. NaHCO3 (20.0 mL) was added to the reaction solution, which was then extracted with CH2Cl2 (20.0 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to yield compound Int H (1.25 g, 4.80 mmol). MS (ESI, m / z): 234.9 [M+H] + .

[0362] Step 4: Synthesis of (3S,5R)-3-(benzo[d]thiazol-5-yl)-5-methylmorpholine (Compound Int H Peak 1) and (3R,5S)-3-(benzo[d]thiazol-5-yl)-5-methylmorpholine (Compound Int H Peak 2)

[0363] Compound Int H was separated by SFC (separation conditions; chiral column: DAICEL CHIRALPAKAD (250 mm*30 mm, 10 μm); mobile phase: [CO₂-EtOH (0.1% NH₃H₂O)]; B%: 45%) to afford compound Int H peak 1 (374 mg, Rt = 1.934 min) and compound Int H peak 2 (423 mg, Rt = 2.130 min). MS (ESI, m / z): 235.0 [M+H] + .

[0364] Int H Peak 1: 1 H NMR (400MHz, CD3OD) δ9.24(s,1H),8.15(s,1H),8.04(d,J=8.4Hz,1H),7.56(dd,J=8.4,1.1Hz,1H),4.13(dd,J=10.4,3.1Hz,1H),3.88 (dd,J=11.2,3.1Hz,1H),3.81(dd,J=10.8,2.7Hz,1H),3.35–3.41(m,1H),3.18–3.23(m,1H),3.10–3.14(m,1H),1.08(d,J=6.4Hz,3H).

[0365] Int H Peak 2: 1 H NMR (400MHz, CD3OD) δ9.25(s,1H),8.16(d,J=0.6Hz,1H),8.05(d,J=8.3Hz,1H),7.57(dd,J=8.4,1.5Hz,1H),4.13(dd,J=10.4,3.2 Hz,1H),3.89(dd,J=11.1,3.3Hz,1H),3.79–3.84(m,1H),3.39(s,1H),3.18–3.25(m,1H),3.09–3.16(m,1H),1.09(d,J=6.4Hz,3H).

[0366] Intermediate Int I: (3R,5S)-3-methyl-5-(5-(trifluoromethyl)pyridin-2-yl)morpholine

[0367] Step 1: Synthesis of 5-methylmorpholin-3-one (Compound Int I-3)

[0368] NaH (120 g, 3.00 mol) was slowly added to THF (1.00 L), and then Int I-1 (100 g, 1.33 mol) was slowly added dropwise in an ice bath. After reacting at 0°C for 30 min, the temperature was raised to 25°C, and Int I-2 (204 g, 1.66 mol) was added to the reaction solution. The reaction was continued at 25°C for 16 hrs. TLC (PE / EtOAc = 1 / 1, R f =0.15) and LCMS showed that the reaction was complete. Saturated NH4Cl solution (1.00 L) was slowly added to quench the reaction. The combined organic phases were extracted with ethyl acetate (6.00 L), dried over anhydrous Na2SO4, filtered, and concentrated to obtain compound Int I-3 (158 g). MS (ESI, m / z): 116.3 [M+H] + .

[0369] Step 2: Synthesis of tert-butyl 3-methyl-5-oxomorpholine-4-carboxylate (Compound Int I-4)

[0370] Compound Int I-3 (158 g, 1.37 mol) was dissolved in THF (1 L), followed by the addition of DMAP (16.8 g, 137 mmol) and Boc2O (329 g, 1.51 mol). The reaction was allowed to proceed at 25°C for 16 hr. TLC (PE / EtOAc = 5 / 1, Rf = 0.35) and LCMS confirmed the completion of the reaction. The product was then concentrated by vacuum distillation and purified by silica gel column chromatography (PE / EtOAc = 1 / 0 to 4 / 1) to afford compound Int I-4 (54.9 g). MS (ESI, m / z): 160.0 [M-55] + .

[0371] 1 H NMR (400MHz, CDCl3) δ4.31-4.14 (m, 3H), 3.81-3.79 (m, 2H), 1.56 (s, 9H), 1.40 (d, J = 6.4Hz, 3H).

[0372] Step 3: Synthesis of tert-butyl 5-((diphenoxyphosphoryl)oxy)-3-methyl-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound Int I-5)

[0373] Compound Int I-4 (54.9 g, 255 mmol) was dissolved in THF (800 mL), replaced with nitrogen, and cooled to -70°C. LiHMDS (1 M, 281 mL) was then slowly added dropwise at -70°C. After the addition was complete, the mixture was stirred at -70°C for 1 hour. Int F-5 (71.9 g, 268 mmol) was then added dropwise to the reaction mixture, and stirred at -60 to -30°C for 3 hours. TLC (PE / EtOAc = 3 / 1, R f =0.34) to detect the occurrence of the reaction, and H2O (200 mL) was added under ice bath to quench the reaction, followed by extraction with EtOAc (300 mL x 3). The organic phases were combined, washed with saturated brine (600 mL), and then dried over anhydrous sodium sulfate, filtered, concentrated, and separated and purified by silica gel column chromatography (PE / EtOAc = 1 / 0 to 9 / 1) to obtain compound Int I-5 (98.9 g).

[0374] Step 4: Synthesis of tert-butyl 3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound Int I-6)

[0375] Compound Int I-5 (98.9 g, 221 mmol), B2Pin2 (84.2 g, 332 mmol), KOAc (65.1 g, 663 mmol), and XPhos-Pd-G3 (18.7 g, 22.1 mmol) were dissolved in 1,4-dioxane (1000 mL). After replacing the atmosphere with nitrogen, the reaction temperature was raised to 100°C and stirred for 2 hours. TLC (PE / EtOAc = 5 / 1, Rf = 0.47) confirmed complete conversion. The mixture was cooled to room temperature and filtered. The filter cake was washed with DCM (1 L). The combined filtrates were concentrated by vacuum distillation and purified by silica gel column chromatography (PE / EtOAc = 1 / 0 to 9 / 1) to afford compound Int I-6 (61.5 g).

[0376] Step 5: Synthesis of tert-butyl 3-methyl-5-(5-(trifluoromethyl)pyridin-2-yl)-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound Int I-7)

[0377] Compound Int I-6 (30.0 g, 92.3 mmol), Int D-8 (20.9 g, 92.3 mmol), LiCl (7.04 g, 166 mmol), Na2CO3 (2 M, 415 mL) and Pd(PPh3)4 (10.7 g, 9.22 mmol) were dissolved in DME (750 mL). After nitrogen replacement, the reaction temperature was raised to 80°C and stirred for 12 hrs. TLC (PE / EtOAc = 5 / 1, R f =0.33) and LCMS detected the formation of the product. After cooling to room temperature, the mixture was concentrated by distillation under reduced pressure, then extracted with water (300 mL) and CH2Cl2 (300 mL x 3). The organic phase was washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and separated and purified by silica gel column chromatography (PE / EtOAc = 1 / 0 to 85 / 15) to obtain compound Int I-7 (11.9 g). MS (ESI, m / z): 344.9 [M+H] + .

[0378] 1 H NMR (400MHz, CDCl3) δ8.73(s,1H),7.81(dd,J=8.4,2.0Hz,1H),7.23(d,J=8.4Hz,1H),6.86(s,1 H), 4.73-4.69 (m, 1H), 4.16-4.13 (m, 1H), 4.04-4.01 (m, 1H), 1.26 (d, J = 7.2Hz, 3H), 1.18 (s, 9H).

[0379] Step 6: Synthesis of 3-methyl-5-(5-(trifluoromethyl)pyridin-2-yl)-3,6-dihydro-2H-1,4-oxazine (Compound Int I-8)

[0380] Compound Int I-5 (8.40 g, 24.4 mmol) was dissolved in TFA (55.6 g, 488 mmol, 36.2 mL) and reacted at 25°C for 1 hour. LCMS confirmed the reaction was complete and the mixture was concentrated to give compound Int I-8 (5.96 g). MS (ESI, m / z): 244.9 [M+H] + .

[0381] Step 7: Synthesis of (3R,5S)-3-methyl-5-(5-(trifluoromethyl)pyridin-2-yl)morpholine (Compound Int I)

[0382] Compound Int I-8 (70.0 mg, 287 μmol) was dissolved in THF (5 mL), and then NaBH3CN (72.1 mg, 1.15 mmol) was added and stirred at 25°C for 5 hrs. LCMS confirmed complete conversion of the reaction, and the mixture was quenched by adding H2O (40 mL). After extraction with CH2Cl2 (600 mL), the organic phases were combined, washed with saturated brine (400 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (CH2Cl2 / MeOH = 100 / 0 to 95 / 5, containing 0.1% TFA, R f =0.33 & 0.24) were separated and purified, concentrated, and then separated by prep-SFC (chiral column: DAICEL CHIRALPAK AD (250mm*30mm, 10um); mobile phase: [CO2-MeOH]; B%: 15%) to obtain compound Int I (3.67g, Rt=0.609min). MS (ESI, m / z): 246.9 [M+H] + .

[0383] 1 H NMR(400MHz, CDCl3)δ8.92-8.86(m,1H),8.09-8.00(m,1H),7.58-7.48(m,1H) ,4.53-4.38(m,1H),4.24-4.02(m,2H),3.68-3.42(m,4H),1.50-1.38(m,3H).

[0384] Example 1: N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(2-(benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl)-2-oxo-acetamide (Compound 1)

[0385] Step 1: Synthesis of 2-(2-(benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl)-N-(7-((2,4-dimethoxybenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-oxoacetamide (Compound 1-1)

[0386] Compounds Int B (300 mg, 1.37 mmol) and Int E (440 mg, 1.14 mmol) were added to a reaction flask and dissolved in anhydrous DMF (4 mL). DIPEA (110 mg, 1.71 mmol) and HATU (520 mg, 1.37 mmol) were then added. The reaction system was incubated at 25°C for 2 hours. After completion, the reaction was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The crude product was isolated and purified by Prep-TLC (DCM:MeOH = 10:1) to obtain compound 1-1 (300 mg). MS (ESI, m / z): 600.2 [M+H] + .

[0387] Step 2: Synthesis of N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(2-(benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl)-2-oxo-acetamide (Compound 1)

[0388] Compound 1-1 (300 mg, 500 μmol) was dissolved in DCM (2 mL) and added to a 4N HCl solution in 1,4-dioxane (2 mL). The reaction system was allowed to react at 25°C for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure and dissolved in an appropriate amount of methanol. A few drops of NH3.H2O were added to adjust the pH to 7-8. The mixture was concentrated under reduced pressure. The crude product was separated and purified by Prep-HPLC to obtain compound 1 (266 mg). MS (ESI, m / z): 450.1 [M+H] + .

[0389] 1 H NMR (400MHz, DMSO-d6) δ10.83–10.66(m,1H),9.42–9.32(m,1H),8.27–8.07(m,2H),7.99–7.72(m,2H),7.64–7.48(m,1H),6.31– 6.14(m,2H),5.77–5.28(m,1H),4.32–4.20(m,3H),3.52–3.45(m,1H),2.04–1.68(m,3H),1.47–1.18(m,3H),1.14–1.02(m,3H).

[0390] Compound 1 was purified by SFC (alkaline conditions; column: DAICEL CHIRALPAK AS (250 mm*30 mm, 10 μm); mobile phase: [CO2-ACN / EtOH (0.1% NH 3.H2O)]; B%: 60%, isocratic elution mode). Separation afforded a mixture 1 of compound 1 isomer 1 and compound 1 isomer 2 (102 mg) and a mixture 2 of compound 1 isomer 3 and compound 1 isomer 4 (105 mg).

[0391] The mixture 1 (102 mg, 226 μmol) of compound 1 isomer 1 and compound 1 isomer 2 was again separated by SFC (alkaline conditions; column: DAICEL CHIRALCEL OJ (250 mm*30 mm, 10 um); mobile phase: [CO2-i-PrOH]; B%: 40%, isocratic elution mode) to obtain compound 1 isomer 1 (10.1 mg, Rt=1.390 min) and compound 1 isomer 2 (53.2 mg, Rt=1.475 min).

[0392] Compound 1 Isomer 1 1 H NMR (400MHz, CD3OD) δ9.31–9.29(m,1H),8.17–8.10(m,2H),8.04–8.00(m,1H),7.94–7.90(m,1H),7.84–7.78(m,1H),7.65–7.54(m,1H),6.07–5. 65(m,1H),4.38–4.30(m,3H),3.88–3.81(m,1H),2.84–2.67(m,2H),2.4 5–2.09(m,2H),1.89–1.77(m,2H),1.41–1.28(m,2H),0.92–0.82(m,3H). MS(ESI,m / z):450.2[M+H] + .

[0393] Compound 1 Isomer 2: N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-((2R,5S)-2-(benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl)-2-oxoacetamide

[0394] 1H NMR(400MHz,MeOD)δ9.32–9.23(m,1H),8.17–7.80(m,4H),7.77–7.52(m,2H),5.97–5.50(m,1H),4.43–4.27(m,3H) ,3.85–3.74(m,2H),3.57–3.48(m,1H),2.46–2.31(m,2H),2.12–1.87(m,3H),1.56–1.48(m,1H),1.24–1.17(m,3H). MS(ESI,m / z):450.1[M+H] + .

[0395] The mixture 2 of compound 1 isomer 3 and compound 1 isomer 4 (105 mg, 233 μmol) was again subjected to SFC (alkaline conditions; column: DAICEL CHIRALCEL OJ-H (250 mm*30 mm, 5 μm); mobile phase: [CO2-i-PrOH (0.1% NH 3. H2O)]; B%: 30%, isocratic elution mode) to separate compound 1 isomer 3 (2.39 mg, Rt = 1.820 min) and compound 1 isomer 4 (46.0 mg, Rt = 2.305 min).

[0396] Compound 1 Isomer 3

[0397] 1 H NMR (400MHz, MeOD) δ9.21–9.12(m,1H),8.06–7.98(m,2H),7.93–7.67(m,2H),7.54–7.41(m,1H),7.30–7.11(m,1H),5.95–5.45(m,1H),4.30–4. 19(m,3H),3.73–3.69(m,1H),3.57–3.52(m,1H),3.39–3.34(m,1H),2.7 2–2.57(m,2H),1.76–1.67(m,2H),1.57–1.44(m,2H),1.32–1.22(m,3H). MS(ESI,m / z):450.2[M+H] + .

[0398] Compound 1 Isomer 4

[0399] 1H NMR (400MHz, MeOD) δ9.31–9.23(m,1H),8.16–7.80(m,4H),7.75–7.50(m,2H),5.96–5.53(m,1H),4.43–4.29(m,3H),3.87–3. 74(m,1H),3.56–3.48(m,1H),2.47–2.28(m,2H),2.09–1.89(m,3H),1.57–1.47(m,1H),1.37–1.27(m,1H),1.25–1.15(m,3H). MS(ESI,m / z):450.1[M+H] + .

[0400] Example 2: N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(5-methyl-2-(5-(trifluoromethyl)pyridin-2-yl)piperidin-1-yl)-2-oxoacetamide (Compound 2)

[0401] Step 1: Synthesis of N-(7-((2,4-dimethoxybenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(5-methyl-2-(5-(trifluoromethyl)pyridin-2-yl)piperidin-1-yl)-2-oxoacetamide (Compound 2-1)

[0402] Compounds Int B (40 mg, 72.66 μmol, 70% purity) and Int D (17.75 mg, 72.66 μmol) were dissolved in anhydrous DMF (1 mL). DIPEA (18.78 mg, 145.31 μmol) and HATU (33.15 mg, 87.19 μmol) were added, and the reaction system was incubated at 25°C for 1 hour. After completion, the reaction was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The crude product was separated and purified by Prep-TLC (DCM:MeOH = 10:1) to obtain compound 2-1 (9 mg). MS (ESI, m / z): 612.3 [M+H] + .

[0403] Step 2: Synthesis of N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(5-methyl-2-(5-(trifluoromethyl)pyridin-2-yl)piperidin-1-yl)-2-oxoacetamide (Compound 2)

[0404] Compound 2-1 (10 mg, 16.35 μmol) was dissolved in DCM (3 mL) and added to a 4N HCl solution in 1,4-dioxane (1 mL). The reaction system was allowed to react at 25°C for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure and dissolved in an appropriate amount of methanol. A few drops of TEA were added to adjust the pH to 7-8, and the mixture was concentrated under reduced pressure. The crude product was separated and purified by Prep-HPLC to obtain compound 2 (3 mg). MS (ESI, m / z): 462.1 [M+H] + .

[0405] 1 H NMR (400MHz, DMSO-d6) δ10.79–10.70(m,1H),9.02–8.98(m,1H),8.30–8. 26(m,1H),7.95–7.86(m,1H),7.77–7.57(m,1H),6.27–6.23(m,2H),5.68– 5.31(m,1H),4.28–4.24(m,3H),3.54–5.50(m,1H),2.45–2.44(m,1H),2. 21–1.86(m,4H),1.64–1.62(m,1H),1.37–1.32(m,1H),1.08–1.06(m,3H).

[0406] The following compounds were prepared by the method and general steps described in Example 2. The other required raw materials can be purchased commercially or synthesized by experienced synthesizers in the field of organic synthesis using conventional reactions from commercially purchased reagents.

[0407] Example 3: N-(4-amino-1-methyl-1H-pyrazolo[4,3-c]pyridin-7-yl)-2-(2-(benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl]-2-oxo-acetamide

[0408] Step 1: Synthesis of 2-[2-(Benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl]-N-(4-((4-methoxyphenyl)methyl)amino)-1-methyl-1H-pyrazolo[4,3-c]pyridin-7-yl]-2-oxo-acetamide (Compound 3-1)

[0409] Int C (20 mg, 42.06 μmol) and Int E (9.77 mg, 42.06 μmol) were dissolved in anhydrous DMF (1 mL). HATU (19.19 mg, 50.47 μmol) and DIPEA (8.15 mg, 63.09 μmol) were added, and the reaction system was incubated at 25°C for 1 hour. After completion, the reaction was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The crude product was separated and purified by Prep-TLC (DCM:MeOH = 10:1) to obtain Intermediate 3-1 (15 mg). MS (ESI, m / z): 690.3 [M+H] + .

[0410] Step 2: Synthesis of N-(4-amino-1-methyl-1H-pyrazolo[4,3-c]pyridin-7-yl)-2-(2-(benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl]-2-oxo-acetamide (Compound 3)

[0411] 3-1 (10 mg, 16.14 μmol) was dissolved in TFA (2 mL), heated to 90°C and reacted for 2 hours. After completion of the reaction, the mixture was concentrated under reduced pressure. The crude product was separated and purified by Prep-HPLC to obtain compound 3 (8 mg). MS (ESI, m / z): 450.1 [M+H] + .

[0412] 1 H NMR (400MHz, DMSO-d6) δ10.53–10.48(m,1H),9.43–9.41(m,1H),8.27–8. 03(m,3H),7.48–7.47(m,1H),6.92–6.81(m,2H),5.78–5.40(m,1H),4.13– 4.08(m,1H),4.04–3.88(m,3H),3.61–3.46(m,1H),2.39–2.13(m,2H),1. 99–1.73(m,2H),1.79–1.73(m,1H),1.45–1.38(m,1H),1.14–1.00(m,3H).

[0413] Example 4: N-(4-amino-1,3-dihydrofuro[3,4-c]pyridin-7-yl)-2-(2-(benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl)-2-oxoacetamide

[0414] Step 1: Synthesis of tert-butyl (7-(2-(2-(benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl)-2-oxoacetamido)-1,3-dihydrofuro[3,4-c]pyridin-4-yl)carbamate (Compound 4-1)

[0415] To the reaction flask, Int E (15 mg, 64.56 μmol), Int A (20 mg, 64.56 μmol), HATU (30 mg, 77.47 μmol), and DMF (2 mL) were added and stirred to dissolve. DIPEA (13 mg, 96.84 μmol) was then added. The mixture was stirred under nitrogen and allowed to react at 25°C for 1 hr. After completion of the reaction, the reaction solution was diluted with water and extracted with ethyl acetate (30 ml x 3). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (DCM:MeOH = 95:5) to obtain compound 4-1 (10 mg). MS (ESI, m / z): 538.3 [M+H] + .

[0416] Step 2: Synthesis of N-(4-amino-1,3-dihydrofuro[3,4-c]pyridin-7-yl)-2-(2-(benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl)-2-oxoacetamide (Compound 4)

[0417] 4-1 (10 mg, 18.60 μmol), TFA (1.5 mL), and DCM (1.5 mL) were added to the reaction flask under nitrogen protection and reacted at 25°C for 2 hours. After the reaction was completed, the reaction solution was spin-dried, diluted with saturated NaHCO3 solution, and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by Pre-HPLC to obtain compound 4 (4.26 mg). MS (ESI, m / z): 438.2 [M+H] + .

[0418] 1H NMR(400MHz,DMSO-d6)δ10.62–10.47(m,1H),9.47–9.38(m,1H),8.25–8.15(m,1H),8.0 9–8.00(m,1H),7.91–7.76(m,1H),7.57–7.41(m,1H),6.07–5.95(m,2H),5.77–5.24(m,1 H),4.94–4.67(m,4H),3.47–3.39(m,1H),3.32–3.29(m,1H),2.40–2.31(m,1H),2.22–2 .04(m,1H),1.96–1.85(m,1H),1.79–1.67(m,1H),1.44–1.31(m,1H),1.10–1.01(m,3H).

[0419] Example 25: 2-(2-(2-(1-acetylpiperidin-4-yl)benzo[d]thiazol-5-yl)-5-methylpiperidin-1-yl)-N-(7-amino-1-methyl-1H-)pyrazolo[3,4-c]pyridin-4-yl)-2-oxoacetamide

[0420] Compound 25-1 (51.0 mg, 70.4 μmol) was dissolved in DCM (5.00 mL), followed by the addition of DDQ (20.8 mg, 91.5 μmol). The reaction was stirred at 20°C for 2 hours. After completion of the reaction as monitored by LCMS, the mixture was filtered and the filter cake was washed twice with dichloromethane. The combined organic phases were concentrated, purified by prep-HPLC, and lyophilized to afford compound 25 (12.0 mg). MS (ESI, m / z): 575.2 [M+H] + .

[0421] 1 H NMR(400MHz, CDCl3)δ9.43(s,1H),8.08(s,1H),7.96–7.86(m,3H),7.39–7.33(m,1H),5.35–4.91(m,3H),4.40–4.33(m,3H),4.32–3.94(m ,1H),3.47–3.09(m,3H),2.87–2.80(m,1H),2.34–2.03(m,2H),2.14(s,3H),2.05–1.84(m,7H),1.49–1.36(m,2H),1.14(d,J=6.4Hz,3H).

[0422] Example 35: N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-((3R,5S)-3-methyl-5-(5-(trifluoromethyl)pyridin)-2-yl)morpholino)-2-oxoacetamide

[0423] Step 1: Synthesis of N-(7-((3,4-dimethylbenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-((3R,5S)-3-methyl-5-(5-(trifluoromethyl)pyridin-2-yl)morpholinyl)-2-oxoacetamide (Compound 35-1)

[0424] Int B (1.02 g, 2.65 mmol) was dissolved in CH2Cl2 (48.0 mL) and THF (8.00 mL), and then DMF (38.7 mg, 529 μmol) and (COCl)2 (840 mg, 6.62 mmol) were added. The reaction was stirred at 20°C for 1 hour. Int I (931 mg, 3.78 mmol) was dissolved in CH2Cl2 (40.0 mL), and DIEA (1.95 g, 15.1 mmol) was added to the reaction mixture and stirred at 25°C for 2 hours. TLC (CH2Cl2 / MeOH = 15 / 1, R f =0.47) and LCMS detected the formation of the product, and after concentration, it was purified by silica gel column chromatography (CH2Cl2 / MeOH=100 / 0 to 90 / 10) to obtain compound 35-1 (1.11 g). MS (ESI, m / z): 614.1 [M+H] + .

[0425] Step 2: Synthesis of N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-((3R,5S)-3-methyl-5-(5-(trifluoromethyl)pyridin)-2-yl)morpholino)-2-oxoacetamide (Compound 35)

[0426] Compound 35-1 (1.11 g, 1.81 mmol) was dissolved in 4M HCl in 1,4-dioxane (13.6 mL) and reacted at 25°C for 1 hr. LCMS confirmed complete conversion. The pH was adjusted to approximately 8 with saturated NaHCO₃ solution, followed by extraction with DCM (120 mL). The combined organic phases were washed with saturated brine (50.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The product was purified by silica gel column chromatography (CH₂Cl₂ / MeOH = 1 / 0 to 19 / 1) and then resolved by prep-SFC (chiral column: DAICEL CHIRALPAK IK (250 mm*30 mm, 10 μm); mobile phase: [CO₂-EtOH (0.1% NH₃H₂O)]; B%: 30%) to afford compound 35 (284 mg, Rt = 1.763 min). MS (ESI, m / z): 464.1 [M+H] + .

[0427] 1 H NMR (400MHz, CDCl3) δ9.42–9.33(m,1H),8.92–8.83(m,1H),8.13–7.89(m,3H),7.73–7.55(m,1H),6.59–5. 76(m,1H),5.23–4.86(m,3H),4.46–4.37(m,4H),4.10–3.92(m,1H),3.89–3.80(m,2H),1.14–0.80(m,3H).

[0428] Example 46: N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(3-methyl-5-(6-(trifluoromethyl)pyridin-3-yl)morpholino)-2-oxoacetamide

[0429] Step 1: Synthesis of tert-butyl 3-methyl-5-(6-(trifluoromethyl)pyridin-3-yl)-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound 46-2)

[0430] Compound Int F (700 mg, 1.56 mmol) and compound 46-1 (427 mg, 1.56 mmol) were dissolved in a mixed solvent of 1,4-dioxane (6 mL) and H2O (2 mL). Pd(PPh3)4 (180 mg, 156 μmol) and K3PO4 (996 mg, 4.69 mmol) were then added sequentially under a nitrogen atmosphere. The reaction was stirred at 90°C for 2 hrs. LCMS detected the formation of the product. The reaction solution was concentrated and purified by silica gel column chromatography (PE / EtOAc = 10 / 1) to afford compound 46-2 (506 mg). MS (ESI, m / z): 345.0 [M+H] + .

[0431] Step 2: Synthesis of 3-methyl-5-(6-(trifluoromethyl)pyridin-3-yl)-3,6-dihydro-2H-1,4-oxazine (Compound 46-3)

[0432] Compound 46-2 (510 mg, 1.49 mmol) was dissolved in CH2Cl2 (5 mL), and then a 2M HCl solution in 1,4-dioxane (2 mL) was added. The reaction was stirred at 25°C for 12 hrs. LCMS confirmed the reaction was complete, and the mixture was concentrated under reduced pressure at 25°C to afford compound 46-3 (361 mg).

[0433] Step 3: Synthesis of 3-methyl-5-(6-(trifluoromethyl)pyridin-3-yl)morpholine (Compound 46-4)

[0434] Compound 46-3 (350 mg, 1.43 mmol) was dissolved in THF (5 mL), and NaBH3CN (360 mg, 5.73 mmol) was added at 0°C. After addition, the reaction was stirred at 25°C for 0.5 hr. LCMS confirmed complete conversion of the starting material. The mixture was concentrated by distillation under reduced pressure to yield compound 46-4 (160 mg). MS (ESI, m / z): 247.0 [M+H] + .

[0435] Step 4: Synthesis of ethyl 2-(3-methyl-5-(6-(trifluoromethyl)pyridin-3-yl)morpholino)-2-oxoacetate (Compound 46-5)

[0436] Compound 46-4 (150 mg, 609 μmol) was dissolved in CH2Cl2 (1 mL), and then DIEA (393 mg, 3.05 mmol, 530 μL) and ethyl oxalyl chloride (249 mg, 1.83 mmol, 204 μL) were added at 0°C, and the reaction was stirred at 25°C for 0.5 hr. LCMS and TLC (CH2Cl2 / MeOH=10 / 1, R f=0.45) to detect the completion of the reaction. The reaction solution was concentrated and then subjected to silica gel column chromatography (CH2Cl2 / MeOH=100 / 1 to 10 / 1, R f =0.45) was separated and purified, and concentrated to give compound 46-5 (210 mg). MS (ESI, m / z): 347.0 [M+H] + .

[0437] Step 5: Synthesis of 2-(3-methyl-5-(6-(trifluoromethyl)pyridin-3-yl)morpholino)-2-oxoacetic acid (Compound 46-6)

[0438] Compound 46-5 (210 mg, 594 μmol) was dissolved in a mixture of THF (3 mL) and H₂O (1 mL). LiOH·H₂O (49.8 mg, 1.19 mmol) was then added and the reaction was stirred at 25°C for 1 hour. After completion of the reaction as determined by LCMS, the pH was adjusted to 2-3 with 1M aqueous hydrochloric acid. The mixture was then extracted with CH₂Cl₂ (20 mL x 3). The organic phase was dried over anhydrous Na₂SO₄, filtered, and concentrated to yield compound 46-6 (163 mg). MS (ESI, m / z): 319.0 [M+H] + .

[0439] Step 6: Synthesis of N-(7-((2,4-dimethylbenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(3-methyl-5-(6-(trifluoromethyl)pyridin-3-yl)morpholino)-2-oxoacetamide (Compound 46-7)

[0440] Compound 46-6 (68.0 mg, 207 μmol) and compound Int B-5 (50.0 mg, 159 μmol) were dissolved in CH2Cl2 (2 mL), and EDCI (45.8 mg, 239 μmol), HOBt (32.3 mg, 239 μmol), and DIEA (61.8 mg, 478 μmol, 83.3 μL) were added in sequence. The reaction was stirred at 25°C for 1 hour. LCMS confirmed the completion of the reaction. The reaction solution was quenched by the addition of H2O (20 mL) and extracted with CH2Cl2 (20 mL x 3). The organic phase was collected, dried over anhydrous Na2SO4, filtered, concentrated, and purified by silica gel column chromatography (CH2Cl2 / MeOH = 100 / 1 to 10 / 1, R f =0.40) was separated and purified to obtain compound 46-7 (43.0 mg). MS (ESI, m / z): 614.1 [M+H] + .

[0441] Step 7: Synthesis of N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(3-methyl-5-(6-(trifluoromethyl)pyridin-3-yl)morpholino)-2-oxoacetamide (Compound 46)

[0442] Compound 46-6 (43.0 mg, 70.0 μmol) was dissolved in CH2Cl2 (1 mL), and a 2M HCl solution in 1,4-dioxane (2 mL) was added. The reaction was stirred at 25°C for 2 hours. After completion of the reaction by LCMS, the mixture was concentrated by vacuum distillation, purified by prep-HPLC, and lyophilized to yield compound 46 (4.86 mg). MS (ESI, m / z): 463.9 [M+H] + .

[0443] 1 H NMR(400MHz,CD3OD)δ8.99(s,1H),8.27–8.35(m,1H),7.99(s,1H),7.78–7.92(m,2H),5.63–5 .83(m,1H),4.79(s,1H),4.36(s,3H),4.18(d,J=1.0Hz,1H),3.84–3.97(m,3H),1.02(s,3H).

[0444] Example 47: (±)-N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(cis-3-methyl-5-(4-(trifluoromethyl)phenyl))morpholino)-2-oxoacetamide (Compound 47) and (±)-N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(trans-3-methyl-5-(4-(trifluoromethyl)phenyl))morpholino)-2-oxoacetamide (Compound 48)

[0445] Step 1: Synthesis of tert-butyl 3-methyl-5-(4-(trifluoromethyl)phenyl)-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound 47-2)

[0446] Compound Int F (2.00 g, 4.47 mmol) and compound 47-1 (1.22 g, 4.47 mmol) were dissolved in a mixed solvent of 1,4-dioxane (12 mL) and H2O (4.00 mL). Pd(PPh3)4 (516 mg, 447 μmol) and K3PO4 (2.85 g, 13.4 mmol) were then added sequentially under a nitrogen atmosphere. The reaction was stirred at 100°C for 12 hrs. TLC (PE / EtOAc = 10 / 1, R f=0.29) to detect the formation of the product. After the reaction solution was concentrated, it was purified by silica gel column chromatography (PE / EtOAc=10 / 1, R f =0.29) was separated and purified, and concentrated to give compound 47-2 (1.50 g). MS (ESI, m / z): 287.8 [M+H- t Bu] + .

[0447] Step 2: Synthesis of 3-methyl-5-(4-(trifluoromethyl)phenyl)-3,6-dihydro-2H-1,4-oxazine (Compound 47-3)

[0448] Compound 47-2 (1.50 g, 4.37 mmol) was dissolved in CH2Cl2 (20 mL), and then a 2M HCl solution in 1,4-dioxane (10 mL) was added. The reaction was stirred at 25°C for 1 hour. LCMS confirmed the reaction was complete, and the mixture was concentrated under reduced pressure at 25°C to afford compound 46-3 (361 mg). MS (ESI, m / z): 262.0 [M+18] + .

[0449] Step 3: Synthesis of 3-methyl-5-(4-(trifluoromethyl)phenyl)morpholine (Compound 47-4)

[0450] Compound 46-3 (1.00 g, 4.11 mmol) was dissolved in MeOH (2.00 mL), and NaBH4 (516 mg, 8.22 mmol) was added at 0°C. After the addition was complete, the reaction was stirred at 25°C for 0.15 hr. LCMS confirmed complete conversion of the starting material. The reaction solution was then extracted with CH2Cl2 (20.0 mL x 3) and the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (CH2Cl2 / MeOH = 10 / 1, R f =0.32) was separated and purified, and concentrated to give compound 47-7 (270 mg). MS (ESI, m / z): 245.9 [M+H] + .

[0451] Step 4: Synthesis of ethyl 2-(3-methyl-5-(4-(trifluoromethyl)phenyl)morpholino)-2-oxoacetate (Compound 47-5)

[0452] Compound 47-4 (270 mg, 671 μmol) was dissolved in CH2Cl2 (0.5 mL), and DIEA (433 mg, 3.36 mmol, 584 μL) and ethyl oxalyl chloride (275 mg, 2.01 mmol, 225 μL) were added at 0°C. The reaction was stirred at 25°C for 0.5 hr. LCMS and TLC (CH2Cl2 / MeOH = 10 / 1, Rf = 0.45) indicated that the reaction was complete. The reaction solution was concentrated and subjected to silica gel column chromatography (CH2Cl2 / MeOH = 100 / 1 to 10 / 1, Rf = 0.45). f =0.45) was separated and purified, and concentrated to give compound 47-5 (380 mg). MS (ESI, m / z): 346.0 [M+H] + .

[0453] Step 5: Synthesis of 2-(3-methyl-5-(4-(trifluoromethyl)phenyl)morpholino)-2-oxoacetic acid (Compound 47-6)

[0454] Compound 47-5 (380 mg, 550 μmol) was dissolved in a mixture of THF (3 mL) and H₂O (1 mL). LiOH·H₂O (46.1 mg, 1.10 mmol) was then added and the reaction stirred at 25°C for 1 hour. After completion of the reaction as determined by LCMS, the pH was adjusted to 2-3 with 1M aqueous hydrochloric acid. The mixture was then extracted with CH₂Cl₂ (20.0 mL x 3). The organic phase was dried over anhydrous Na₂SO₄, filtered, and concentrated to yield compound 47-6 (349 mg). MS (ESI, m / z): 317.8 [M+H] + .

[0455] Step 6: Synthesis of N-(7-((2,4-dimethylbenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(3-methyl-5-(4-(trifluoromethyl)phenyl)morpholinyl)-2-oxoacetamide (Compound 47-7)

[0456] Compound 47-6 (87.0 mg, 191 μmol) and compound Int B-5 (50.0 mg, 159 μmol) were dissolved in CH2Cl2 (2 mL), and EDCI (45.8 mg, 239 μmol), HOBt (32.3 mg, 239 μmol), and DIEA (61.8 mg, 478 μmol, 83.3 μL) were added in sequence. The reaction was stirred at 25°C for 1 hour. LCMS confirmed the completion of the reaction. The reaction solution was quenched by the addition of H2O (20 mL) and extracted with CH2Cl2 (20.0 mL x 3). The organic phase was collected, dried over anhydrous Na2SO4, filtered, concentrated, and purified by silica gel column chromatography (CH2Cl2 / MeOH = 100 / 1 to 10 / 1, R f =0.45) was separated and purified to obtain compound 47-7 (65.0 mg). MS (ESI, m / z): 613.0 [M+H] + .

[0457] Step 7: Synthesis of (±)-N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(cis-3-methyl-5-(4-(trifluoromethyl)phenyl))morpholino)-2-oxoacetamide (Compound 47) and (±)-N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(trans-3-methyl-5-(4-(trifluoromethyl)phenyl))morpholino)-2-oxoacetamide (Compound 48)

[0458] Compound 47-6 (65.0 mg, 50.9 μmol) was dissolved in CH2Cl2 (4 mL), and a 2M HCl solution in 1,4-dioxane (2 mL) was added. The reaction was stirred at 25°C for 2 hours. After completion of the reaction by LCMS, the product was concentrated by distillation under reduced pressure, separated and purified by prep-HPLC, and lyophilized to afford compound (±)47 (3.83 mg) (Rt = 1.466 min) and compound (±)48 (3.56 mg) (Rt = 1.246 min). MS (ESI, m / z): 462.9 [M+H] + .

[0459] Compound (±) 47: 1 H NMR (400MHz, CD3OD) δ8.39–8.49(m,1H),7.43–8.02(m,6H),5.68(d,J=1.2Hz,1H) ,4.79(s,1H),4.58(s,1H),4.35(s,3H),3.83–4.13(m,3H),1.00(d,J=5.6Hz,3H).

[0460] Compound (±) 48: 1 H NMR (400MHz, CD3OD) δ8.29–8.44(m,1H),7.73–7.90(m,1H),7.34–7.70(m,5H),5.17–5.23(m,1H),4.55–4. 57(m,1H),4.27–4.36(m,3H),3.91–4.00(m,2H),3.68–3.84(m,1H),3.50–3.64(m,1H),1.43–1.59(m,3H).

[0461] The following compounds were prepared by the method and general steps described in Example 47. The other required raw materials can be purchased commercially or synthesized by experienced synthesizers in the field of organic synthesis using conventional reactions from commercially purchased reagents.

[0462] Example 49: N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-((2R,5S)-5-methyl-2-(2-(1-methylpiperidin)-4-yl)benzo[d]thiazol-5-yl)piperidin-1-yl)-2-oxoacetamide

[0463] Step 1: Synthesis of N-(7-((2,4-dimethylbenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-((2R,5S)-5-methyl-2-(2-(1-methylpiperidin-4-yl)benzo[d]thiazol-5-yl)piperidin-1-yl)-2-oxoacetamide (Compound 49-2)

[0464] Compound 49-1 (90.0 mg, 273.14 μmol) and Int B (105 mg, 273 μmol) were dissolved in DMF (5 mL), followed by the addition of DIEA (106 mg, 819 μmol, 143 μL) and HATU (156 mg, 410 μmol). The reaction was stirred at 25°C for 1 hour. LCMS confirmed the reaction was complete, followed by quenching with H2O (30.0 mL) and extraction with EtOAc (30 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to yield compound 49-2 (150 mg). MS (ESI, m / z): 697.2 [M+H] + .

[0465] Step 2: Synthesis of N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-((2R,5S)-5-methyl-2-(2-(1-methylpiperidin)-4-yl)benzo[d]thiazol-5-yl)piperidin-1-yl)-2-oxoacetamide (Compound 49)

[0466] Compound 49-2 (150 mg, 215 μmol) was dissolved in TFA (5 mL) and heated to 80°C with stirring for 0.5 hr. LCMS confirmed the reaction was complete. Saturated NaHCO₃ (50 mL) was added to the reaction solution, which was then extracted with CH₂Cl₂ (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The product was then purified by prep-HPLC and lyophilized to yield compound 49 (43.57 mg). MS (ESI, m / z): 547.2 [M+H] + .

[0467] 1 H NMR (400MHz, DMSO-d6) δ10.65–10.89(m,1H),8.04–8.13(m,1H),7.95–8.01(m,1H),7.92(br s,2H),7.36–7.52(m,1H),6.16–6.29(m,2H),5.22–5.81(m,1H),4.20–4.37(m,3H),3.42–3.50(m,2H),2.81 –3.14(m,4H),2.20(s,3H),1.98–2.15(m,5H),1.71–1.94(m,4H),1.33–1.44(m,1H),1.08(d,J=7.1Hz,3H).

[0468] Example 50: N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(3-methyl-5-(5-(trifluoromethoxy)pyridin-2-yl)morpholino)-2-oxoacetamide

[0469] Step 1: Synthesis of tert-butyl 3-methyl-5-(5-(trifluoromethoxy)pyridin-2-yl)-2,3-dihydro-4H-1,4-oxazine-4-carboxylate (Compound 50-2)

[0470] Compound Int G (699 mg, 2.15 mmol) and compound 50-1 (400 mg, 1.65 mmol) were dissolved in a mixed solvent of 1,4-dioxane (30 mL) and H2O (10 mL). XPhos-Pd-G3 (140 mg, 165 μmol) and K3PO4 (1.05 g, 4.96 mmol) were then added sequentially under a nitrogen atmosphere. The reaction was stirred at 80°C for 12 hours. LCMS analysis revealed the formation of the product. The reaction solution was concentrated and purified by silica gel column chromatography (PE / EtOAc = 100 / 0 to 92 / 8, Rf = 0.49) to afford compound 50-2 (358 mg). MS (ESI, m / z): 360.9 [M+H] + .

[0471] Step 2: Synthesis of 3-methyl-5-(5-(trifluoromethoxy)pyridin-2-yl)-3,6-dihydro-2H-1,4-oxazine (Compound 50-3)

[0472] Compound 50-2 (318 mg, 883 μmol) was dissolved in CH2Cl2 (1 mL), and then a 2M HCl solution in 1,4-dioxane (1 mL) was added. The reaction was stirred at 25°C for 12 hr. LCMS confirmed the reaction was complete, and the mixture was concentrated under reduced pressure at 25°C to afford compound 46-3 (229 mg).

[0473] Step 3: Synthesis of 3-methyl-5-(5-(trifluoromethoxy)pyridin-2-yl)morpholine (Compound 50-4)

[0474] Compound 50-3 (229 mg, 880 μmol) was dissolved in MeOH (5 mL) and THF (20 mL). NaBH3CN (221 mg, 3.52 mmol) was then added at 0°C. After the addition was complete, the reaction was stirred at 25°C for 1.5 hours. LCMS confirmed complete conversion of the starting material. H2O (10 mL) was added to the reaction solution, followed by extraction with CH2Cl2 (20 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, concentrated, and purified by silica gel column chromatography (CH2Cl2 / MeOH = 100 / 0 to 85 / 15, R f =0.35) was separated and purified, and concentrated to give compound 50-4 (158 mg). MS (ESI, m / z): 262.9 [M+H] + .

[0475] Step 4: Synthesis of ethyl 2-(3-methyl-5-(5-(trifluoromethoxy)pyridin-2-yl)morpholino)-2-oxoacetate (Compound 50-5)

[0476] Compound 50-4 (158 mg, 603 μmol) was dissolved in CH2Cl2 (10 mL), and then DIEA (311 mg, 2.41 mmol, 420 μL) and ethyl oxalyl chloride (123 mg, 904 μmol, 101 μL) were added at 0°C. The reaction was stirred at 25°C for 1 hour. LCMS confirmed the reaction was complete. The reaction solution was concentrated and subjected to silica gel column chromatography (CH2Cl2 / MeOH = 100 / 0 to 90 / 10, R f =0.65) was separated and purified, and concentrated to give compound 50-5 (193 mg). MS (ESI, m / z): 363.0 [M+H] + .

[0477] Step 5: Synthesis of 2-(3-methyl-5-(5-(trifluoromethoxy)pyridin-2-yl)morpholino)-2-oxoacetic acid (Compound 50-6)

[0478] Compound 50-5 (193 mg, 533 μmol) was dissolved in a mixture of THF (12 mL) and H₂O (4 mL). LiOH·H₂O (44.7 mg, 1.07 mmol) was then added and the reaction stirred at 25°C for 1 hour. After completion of the reaction as determined by LCMS, the pH was adjusted to 2-3 with 1M aqueous hydrochloric acid. The mixture was then extracted with CH₂Cl₂ (20 mL x 3). The organic phase was dried over anhydrous Na₂SO₄, filtered, and concentrated to yield compound 50-6 (186 mg). MS (ESI, m / z): 335.1 [M+H] + .

[0479] Step 6: Synthesis of N-(7-((2,4-dimethylbenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(3-methyl-5-(5-(trifluoromethoxy)pyridin-2-yl)morpholino)-2-oxoacetamide (Compound 50-7)

[0480] Compound 50-6 (186 mg, 556 μmol), compound Int B-5 (174 mg, 556 μmol), and HATU (423 mg, 1.11 mmol) were dissolved in DMF (2 mL), and DIEA (216 mg, 1.67 mmol, 291 μL) was added. The reaction was stirred at 25°C for 1 hour. LCMS confirmed the completion of the reaction. The reaction solution was quenched with H2O (20 mL) and extracted with EA (20 mL x 3). The organic phase was collected, dried over anhydrous Na2SO4, filtered, concentrated, and purified by silica gel column chromatography (CH2Cl2 / MeOH = 100 / 0 to 86 / 14, R f=0.37) to obtain compound 50-7 (239 mg). MS (ESI, m / z): 630.2 [M+H] + .

[0481] Step 7: Synthesis of N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-(3-methyl-5-(5-(trifluoromethoxy)pyridin-2-yl)morpholino)-2-oxoacetamide (Compound 50)

[0482] Compound 50-6 (239 mg, 380 μmol) was dissolved in CH2Cl2 (3 mL), and then a 2M HCl solution in 1,4-dioxane (1 mL) was added. The reaction was stirred at 25°C for 2 hours. After completion of the reaction by LCMS, the mixture was concentrated by vacuum distillation, purified by prep-HPLC, and lyophilized to obtain compound 50 (96.83 mg). MS (ESI, m / z): 479.9 [M+H] + .

[0483] 1 H NMR (400MHz, CDCl3) δ9.32–9.47(m,1H),8.31–8.61(m,1H),7.82–8.19(m,2H),7.45–7.68(m,2H),5.65–6.48(m,2H),4.76–5.21(m,1H),4.40(br s,3H),3.75–4.09(m,3H),0.76–1.14(m,3H).

[0484] Example 51: N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-((3S,5R)-3-(benzo[d]thiazol-5-yl)-5-methylmorpholino)-2-oxoacetamide

[0485] Step 1: Synthesis of ethyl 2-((3S,5R)-3-(benzo[d]thiazol-5-yl)-5-methylmorpholino)-2-oxoacetate (Compound 51-1)

[0486] Compound Int H peak 1 (364 mg, 1.46 mmol) was dissolved in CH2Cl2 (5 mL), and then DIEA (943 mg, 7.30 mmol, 1.27 mL) and ethyl oxalyl chloride (598 mg, 4.38 mmol, 489 μL) were added at 0°C. The reaction was stirred at 25°C for 1 hour. LCMS confirmed the reaction was complete. The reaction solution was concentrated and purified by silica gel column chromatography (CH2Cl2 / MeOH = 10 / 1, R f=0.50) was separated and purified, and concentrated to give compound 51-1 (495 mg). MS (ESI, m / z): 335.0 [M+H] + .

[0487] Step 2: Synthesis of 2-((3S,5R)-3-(benzo[d]thiazol-5-yl)-5-methylmorpholino)-2-oxoacetic acid (Compound 51-2)

[0488] Compound 51-1 (495 mg, 1.20 mmol) was dissolved in a mixture of THF (3 mL) and H₂O (1 mL). LiOH·H₂O (100 mg, 2.40 mmol) was then added and the reaction was stirred at 25°C for 1 hour. After completion of the reaction, the pH was adjusted to 2-3 with 1M aqueous hydrochloric acid, followed by extraction with CH₂Cl₂ (20 mL x 3). The organic phase was dried over anhydrous Na₂SO₄, filtered, and concentrated to yield compound 51-2 (367 mg). MS (ESI, m / z): 307.0 [M+H] + .

[0489] Step 3: Synthesis of 2-((3S,5R)-3-(benzo[d]thiazol-5-yl)-5-methylmorpholino)-N-(7-((2,4-dimethylbenzyl)amino)-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-oxoacetamide (Compound 51-3)

[0490] Compound 51-2 (333 mg, 1.08 mmol) and compound Int B-5 (260 mg, 829 μmol) were dissolved in CH2Cl2 (4 mL), and EDCI (238 mg, 1.24 mmol), HOBt (168 mg, 1.24 mmol), and DIEA (321 mg, 2.49 mmol, 433 μL) were added in sequence. The reaction was stirred at 25°C for 1 hour. LCMS confirmed the reaction was complete, and the product was concentrated and purified by silica gel column chromatography (CH2Cl2 / MeOH = 10 / 1, R f =0.54) was separated and purified to obtain compound 51-3 (400 mg). MS (ESI, m / z): 602.0 [M+H] + .

[0491] Step 4: Synthesis of N-(7-amino-1-methyl-1H-pyrazolo[3,4-c]pyridin-4-yl)-2-((3S,5R)-3-(benzo[d]thiazol-5-yl)-5-methylmorpholino)-2-oxoacetamide (Compound 51)

[0492] Compound 51-3 (400 mg, 602 μmol) was dissolved in CH2Cl2 (10 mL), followed by the addition of a 2M HCl solution in 1,4-dioxane (5 mL). The reaction was stirred at 25°C for 1 hour. After completion of the reaction by LCMS, the mixture was concentrated by vacuum distillation, purified by prep-HPLC, and lyophilized to afford compound 51 (163 mg). MS (ESI, m / z): 452.0 [M+H] + .

[0493] 1 H NMR (400MHz, CD3OD) δ9.26 (s, 1H), 8.42 (s, 1H), 8.08 (d, J = 8.4Hz, 1H), 7.75–8.02 (m,3H),5.76–5.84(m,1H),4.34(s,3H),3.81–4.03(m,5H),1.00(d,J=5.6Hz,3H).

[0494] The following compounds were prepared by the method and general steps described in Example 51. The other required raw materials can be purchased commercially or synthesized by experienced synthesizers in the field of organic synthesis using conventional reactions from commercially purchased reagents.

[0495] Separation method

[0496] The Prep-HPLC purification of the compounds in the examples was performed using an Aglient 1260 or Waters 2489 HPLC, and the separation column model was a Waters SunFire Prep C 18 OBD(19mm×150mm×5.0μm), Waters Xbridge Prep C 18 OBD (19mm×150mm×5.0μm) or YMC Actus Triart C 18 (20 mm × 150 mm × 5.0 μm), the column temperature was 25°C, the detection wavelength was 214 nm, 254 nm, or 280 nm, mobile phase A was acetonitrile, mobile phase B was 0.05% formic acid aqueous solution or 0.05% ammonium bicarbonate aqueous solution or 0.05% TFA aqueous solution, the volume ratio of the mobile phase was adjusted according to the polarity of the compound; the mobile phase flow rate was 28 mL / min.

[0497] Biological evaluation

[0498] Experimental Example 1: Inhibitory effect of test compounds on PRMT5-MTA methyltransferase activity

[0499] The prepared protein solution (PRMT5 / MEP50 and MTA mixture) was preincubated with different concentrations of the test compound (500 nM starting point, 5-fold dilution, 7 points; or 100 nM starting point, 5-fold dilution, 7 points) at 25°C for 30 minutes. The prepared substrate solution (biotinylated histone H4 peptide) was added and incubated at 25°C for 90 minutes. After the reaction, the prepared detection reagent mixture (Protein A-Eu, anti-histone H4 antibody, and Streptavidin-D2) was added and incubated at 25°C for 60 minutes. The fluorescence signal ratio (Ratio) was detected using a BMG microplate reader.

[0500] The solvent group (DMSO) was used as the negative control and the reaction buffer group (without PRMT5·MTA enzyme) was used as the blank control. The percentage inhibition rate of compounds at different concentrations was calculated according to the following formula:

[0501] Percent inhibition rate = (negative control Ratio - compound Ratio) / (negative control Ratio - blank control Ratio) × 100%.

[0502] The four-parameter equation Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC 50 -X)*Hillslope)) fits the detection signal value and calculates IC 50 Where Y is the relative inhibitory activity percentage, Top and Bottom are the maximum and minimum values ​​of the fitted curve, respectively, X is the logarithmic concentration of the compound, and Hillslope is the slope of the curve. The inhibitory effects of the compounds on PRMT5-MTA methyltransferase were determined according to the above method, and the results are shown in Table 1.

[0503] Table 1. Inhibitory activity of the compounds of the present invention against PRMT5-MTA

[0504] The experimental results show that the compounds of the present invention have a strong inhibitory effect on PRMT5-MTA enzyme.

[0505] Experimental Example 2: Inhibitory Effect of Compounds on MTAP Deleted / Parental HCT116 Cell Proliferation

[0506] In this example, MTAP-deficient / parental HCT116 cells were used and purchased from HORIZON.

[0507] MTAP-deficient / parental HCT116 cells were cultured as monolayers in RPMI6140 medium supplemented with 10% FBS and 1% P / S at 37°C and 5% CO2. Logarithmically growing cells were digested and the concentration adjusted. 250 cells per well were seeded into a 96-well plate and cultured overnight. Prediluted compounds were added (5000 nM starting point, 4-fold dilution, 9 points). A negative control group received DMSO, while a blank control group received culture medium. After 8 days of incubation at 37°C and 5% CO2, 50 μl of CellTiter-Glo was added to each well for lysis at room temperature in the dark for 10 minutes. The assay solution was transferred to a 96-well opaque white plate, and the relative chemiluminescence units (RLUs) were read on a microplate reader in chemiluminescence detection mode.

[0508] The percentage inhibition rate of compounds at different concentrations was calculated according to the following formula:

[0509] Percent inhibition rate=(1-(chemiluminescent signal value of the test compound-chemiluminescent signal value of the blank control) / (chemiluminescent signal value of the negative control-chemiluminescent signal value of the blank control))×100%.

[0510] The percentage inhibition rate of different concentrations of compounds was plotted against the compound concentration, and the curve was fitted according to the four-parameter model to calculate the IC 50 value:

[0511] y=Min+(Max-Min) / (1+(x / IC 50 )^(-Hillslope)), where: y is the percentage inhibition rate; Max and Min are the maximum and minimum values ​​of the fitted curve, respectively; x is the logarithmic concentration of the compound; and Hillslope is the slope of the curve.

[0512] The proliferation inhibitory activity of the compounds on MTAP-deficient / parental HCT116 cells was determined according to the above method. The results are shown in Table 2.

[0513] Table 2. Proliferation inhibitory activity of the compounds of the present invention on MTAP-deficient / parental HCT116 cells

[0514] The experimental results show that the compound of the present invention has a strong inhibitory effect on MTAP-deficient HCT116 cells and has a certain selectivity relative to MTAP parent HCT116 cells.

[0515] Experimental Example 3: Pharmacokinetics of the compound in BALB / c mice

[0516] The compounds of the present invention were administered to female Balb / c mice by oral gavage (PO) to investigate their pharmacokinetic characteristics.

[0517] The compound of the present invention and TNG462 (prepared according to patent WO2022026892A1) were administered intravenously (1 mg / kg) and orally (10 mg / kg). The IV solvent was 5% DMSO + 5% Solutol + 90% Saline, and the PO solvent was 10% Solutol + 90% H2O. Blood samples (40-50 μl) were collected intraorbitally at 0 h before dosing and at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 10, and 24 h after dosing (intravenously) and at 0 h before dosing and at 0.25, 0.5, 1, 2, 4, 6, 8, 10, and 24 h after dosing (orally). The samples were placed in K2-EDTA anticoagulant tubes (TNG462 samples were temporarily stored on wet ice) and centrifuged at 4000 rpm for 10 min at 4°C. Plasma was separated (1 μl of 30% formic acid solution was pre-added to the TNG462 plasma tube) and stored at -80°C until analysis. 10 μl of plasma sample was added to 90 μl of acetonitrile containing the internal standard, shaken to mix, and centrifuged at 4000 rpm at 4°C for 10 min. The supernatant was analyzed by LC-MS / MS. WinNonlin 6.3 software was used to calculate the pharmacokinetic parameters using a non-compartmental model. The results are shown in Table 3.

[0518] Table 3: Pharmacokinetic parameters of the compounds in mice

[0519] TNG462 structure:

[0520] in conclusion:

[0521] The above experimental results show that the compound of the present invention has a low body clearance rate, high blood concentration and body exposure, high bioavailability, and good oral absorption effect.

[0522] In addition to those described herein, various modifications of the present invention 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 application (including all patents, patent applications, journal articles, books, and any other disclosures) is incorporated herein by reference in its entirety.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof: in: represents a single bond or a double bond; X 1 , X 2 and X 3 are each independently selected from -CH2-, -CH-, -O-, -N-, -NH- and -S-; R 1 is independently selected at each occurrence from H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl and C 3-8 Cycloalkyl; Ring A is a 5-7 membered saturated or partially saturated heterocyclic ring containing at least one N atom and optionally 1 or 2 other heteroatoms independently selected from O or S which are the same or different; R 2 is independently selected at each occurrence from H, -OH, halogen, -CN, -NR 4 R 5 、-COCH3、-NHCOCH3、C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Hydroxyalkyl, C 2-6 Heteroalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, C 3-8 Cycloalkoxy, C 6-10 Aryl and 5-10 membered heteroaryl, wherein the alkyl, alkoxy, hydroxyalkyl, heteroalkyl, cycloalkyl, heterocyclyl, cycloalkoxy, aryl or heteroaryl are each optionally substituted by one or more R 3 replace; R 3 is independently selected at each occurrence from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, C 3-6 Cycloalkyl, C 3-8 Cycloalkoxy, 3-8 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl, wherein the alkyl, alkoxy, heteroalkyl, cycloalkyl, cycloalkoxy, heterocyclyl, aryl or heteroaryl are each optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 3-8 Substitution of cycloalkoxy and 3-8 membered heterocyclic groups; R 4 , R 5 , R 6 and R 7 Each independently selected from H and C 1-6 Alkyl; or R 4 and R 5 , R 6 and R 7 Together with the nitrogen atom to which it is attached, it forms a 3-8 membered heterocyclic group, wherein the heterocyclic group is optionally substituted by one or more independently selected from halogen, OH, CN, -NH2 and C 1-6 Substitution of alkyl groups; m is 0, 1, 2 or 3; n is 0, 1, 2, 3 or 4.

2. The compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: X 1 , X 2 and X 3 are each independently selected from -CH2-, -CH-, -O-, -N- and -NH-; Preferably, X 1 is -CH-, and X 2 and X 3 for -N-; or Preferably, X 1 and X 2 is -N-, and X 3 is -CH-; or Preferably, X 1 and X 3 is -CH2-, and X 2 For -O-.

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: R 1 Each occurrence is independently selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl and C 3-8 Cycloalkyl; Preferably, R 1 Each occurrence is independently selected from H and C 1-6 alkyl; Preferably, R 1 It is methyl.

4. The compound of any one of claims 1 to 3 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: Ring A is a 5-7 membered saturated heterocyclic ring comprising at least one N atom and optionally 1 or 2 other heteroatoms independently selected from O or S which are the same or different; Preferably, ring A is a 5-7 membered saturated nitrogen heterocycle and a 5-7 membered saturated N and O-containing heterocycle; Preferably, ring A is a 6-membered saturated nitrogen heterocycle and a 6-membered saturated N- and O-containing heterocycle; Preferably, ring A is a piperidine ring and a morpholine ring.

5. The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: R 2 is independently selected at each occurrence from H, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Hydroxyalkyl, C 2-6 Heteroalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, C 3-8 Cycloalkoxy, C 6-10 Aryl and 5-10 membered heteroaryl, wherein the alkyl, alkoxy, hydroxyalkyl, heteroalkyl, cycloalkyl, heterocyclyl, cycloalkoxy, aryl or heteroaryl are each optionally substituted by one or more R 3 replace; Preferably, R 2 is independently selected at each occurrence from H, halogen, C 1-6 Alkyl, C 6-10 Aryl and 5-10 membered heteroaryl, wherein the alkyl, aryl and heteroaryl are each optionally substituted by one or more R 3 replace; Preferably, R 2 Each occurrence is independently selected from C 1-6 Alkyl, C 6-10 Aryl and 6-9 membered heteroaryl, wherein the alkyl, aryl or heteroaryl groups are each optionally substituted by one or more R 3 replace; Preferably, R 2 Each occurrence is independently selected from C 1-6 alkyl, phenyl, pyridyl and benzothiazolyl, wherein the phenyl, pyridyl or benzothiazolyl is each optionally substituted by one or more R 3 replace; Preferably, R 2 is independently selected at each occurrence from methyl, The wavy line It indicates the point of attachment of the group to the rest of the molecule.

6. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: R 3 is independently selected at each occurrence from halogen, -CN, -NR 4 R 5 , C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, 3-8 membered heterocyclic group, C 6-10 Aryl and 5-6 membered heteroaryl, wherein the alkyl, heteroalkyl, heterocyclyl, aryl or heteroaryl are each optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 3-8 Substitution of cycloalkoxy and 3-8 membered heterocyclic groups; Preferably, R 3 is independently selected at each occurrence from halogen, -NR 4 R 5 , C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 2-6 heteroalkyl, 4-7 membered heterocyclyl, phenyl and 5-6 membered heteroaryl, wherein each of the alkyl, heteroalkyl, heterocyclyl, phenyl or heteroaryl is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Substitution of cycloalkyl and 4-6 membered heterocyclic groups; Preferably, R 3 Each occurrence is independently selected from -NR 4 R 5 , C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, 4-7 membered heterocyclyl and 5-6 membered heteroaryl, the alkyl, heterocyclyl, heteroaryl are optionally substituted with one or more -NR 4 R 5 、-COCH3、C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-6 Substitution of cycloalkyl and 4-6 membered heterocyclic groups; Preferably, R 3 is independently selected at each occurrence from methyl, trifluoromethyl, ethyl, amino, methylamino, trifluoromethoxy, The wavy line It indicates the point of attachment of the group to the rest of the molecule.

7. A compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: R 6 and R 7 are each independently selected from H and methyl; Preferably, R 6 and R 7 For H.

8. The compound of any one of claims 1 to 7, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: m is 0 or 1.

9. The compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: n is 2.

10. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula II:

11. The compound of claim 10 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: X 1 is -CH-, and X 2 and X 3 for -N-; R 1 is methyl; and m is 1.

12. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula III:

13. The compound of claim 12 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: X 1 is -CH-, and X 2 and X 3 for -N-; R 1 is methyl; m is 1; and R 3 Selected from methyl, trifluoromethyl, ethyl, amino, methylamino, trifluoromethoxy, The wavy line It indicates the point of attachment of the group to the rest of the molecule.

14. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula IV:

15. The compound of claim 14 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein: X 1 is -CH-, and X 2 and X 3 for -N-; R 1 is methyl; m is 1; and R 3 Selected from methyl, trifluoromethyl, ethyl, amino, methylamino, trifluoromethoxy, The wavy line It indicates the point of attachment of the group to the rest of the molecule.

16. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula V: in, Y 1 , Y 2 are each independently selected from -CH- and -N-.

17. The compound of claim 16 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula V-1:

18. The compound of claim 17 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula V-1a:

19. The compound of claim 17 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula V-1b:

20. The compound of claim 17 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula V-1c:

21. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of Formula VI: in, p is 0 or 1.

22. The compound of claim 21 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula VI-1:

23. The compound of claim 22 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula VI-1a:

24. The compound of claim 22 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is a compound of formula VI-1b:

25. The compound of any one of claims 1-24, or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, wherein the compound is selected from:

26. A pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound according to any one of claims 1 to 25 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotopically labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof and one or more pharmaceutically acceptable carriers.

27. Use of a compound according to any one of claims 1 to 25 or a pharmaceutically acceptable salt, stereoisomer, tautomer, isotope-labeled compound, polymorph, solvate, N-oxide, metabolite or prodrug thereof, or a pharmaceutical composition according to claim 26 in the preparation of a medicament for preventing or treating a disease or condition associated with PRMT5 activity, Preferably, the disease or condition associated with PRMT5 activity is preferably a cancer or tumor with MTAP deficiency; Preferably, the cancer or tumor is esophageal cancer, lung cancer, pancreatic cancer, glioblastoma, bile duct cancer, bladder cancer, breast cancer, ovarian cancer, hepatocellular carcinoma, prostate cancer, melanoma, gastric cancer, colon cancer, leukemia (particularly chronic B-lymphocytic leukemia) or lymphoma.

28. A method for preparing a compound according to any one of claims 1 to 25, comprising the following steps: Step 1: Compound IA-1 and acyl chloride Compound IA-2 is generated through condensation reaction; Step 2: Compound IA-2 is hydrolyzed to generate compound IA-3; Step 3: Compound IA-3 and compound IA-4 undergo condensation reaction to generate compound IA-5; Step 4: removing the protecting group from the condensation product IA-5 to generate compound I; in PG is an amino protecting group, preferably p-methoxybenzyl or 2,4-dimethoxybenzyl; and X 1 , X 2 , X 3 , Ring A, R 1 , R 2 , m, n are as defined in any one of claims 1 to 25, R 6 and R 7 For H.

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