Methylpyrazole compound, pharmaceutical composition containing methylpyrazole compound, and preparation method and application of methylpyrazole compound

Abstract

The invention relates to a methylpyrazole compound, a pharmaceutical composition containing the methylpyrazole compound as well as a preparation method and application of the methylpyrazole compound. Specifically, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt, a stereoisomer, a tautomer or an isotope-labeled compound thereof, a pharmaceutical composition comprising the same, a preparation method thereof, and a use thereof in the preparation of drugs for the prevention or treatment of diseases or conditions related to PRMT5 activity. # imgabs0 #

Description

Methylpyrazole compound, pharmaceutical composition containing the same, preparation method and use thereof

[0001] This application claims priority to Chinese application No. 202210903971.1, filed on July 29, 2022, entitled “Methylpyrazole compounds, pharmaceutical compositions containing the same, preparation methods and uses thereof”, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0002] The present invention relates to methylpyrazole compounds, pharmaceutical compositions containing the same, methods for preparing the same, and their use in preventing or treating diseases or conditions associated with PRMT5 activity. Background Art

[0003] 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).

[0004] 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.).

[0005] 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.

[0006] 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.

[0007] 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.

[0008] Summary of the Invention

[0009] The present invention provides a new methylpyrazole compound, which has a good inhibitory effect on MTAP-deficient tumor cells and has good pharmacokinetic properties.

[0010] One aspect of the present invention provides a compound of Formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof:

[0011] in:

[0012] R 1 for

[0013] R 2 and R 6 are independently selected from H, halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkoxy or 3-8 membered heterocyclic oxy;

[0014] R 3 、R 4 and R 5 are each independently selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution,

[0015] -XLC 3-8Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0016] -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0017] -XLC 6-10 Aryl, said aryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,

[0018] -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,

[0019] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0020] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0021] or R 3 With R 4 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl,

[0022] or R 4 With R 5 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl,

[0023] The condition is that when R 1 for When R 3 、R 4 and R 5 At least one selected from the above -XLC 3-8 Cycloalkyl, -XL-3-8 membered heterocycloalkyl, -XLC 6-10 Aryl, -XL-5-10 membered heteroaryl, -XL-5-10 membered bridged ring group, -XL-5-10 membered spiro ring group, 3-8 membered heterocyclic group;

[0024] X is independently selected at each occurrence from O, S or NR 7 ;

[0025] L is independently a direct bond or -(CH2) n -;

[0026] n is 1 or 2;

[0027] R 7 Selected from H, C 1-4 Alkyl or C 1-4 alkyl halide;

[0028] R 8 and R 9 Independently selected from H, C 1-4 Alkyl or C 1-4 Halogenated alkyl.

[0029] Another aspect of the present invention provides a compound of Formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof:

[0030] in:

[0031] R 1 for

[0032] R 2 and R 6 are independently selected from H, halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 cycloalkoxy;

[0033] R 3 、R 4 and R 5 are each independently selected from H, halogen, C 1-4Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 substituted with a haloalkoxy group or a 3-6 membered heterocyclic group,

[0034] -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0035] -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0036] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0037] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0038] or R 3 With R 4 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl,

[0039] or R 4 With R 5 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl,

[0040] The condition is that when R 1 for When R 3 、R4 and R 5 At least one selected from the above -XLC 3-8 Cycloalkyl, -XL-3-8 membered heterocycloalkyl, -XL-5-10 membered bridged ring group, -XL-5-10 membered spirocyclic group, 3-8 membered heterocyclic group;

[0041] X is independently selected at each occurrence from O, S or NR 7 ;

[0042] L is independently a direct bond or -(CH2) n -;

[0043] n is 1 or 2;

[0044] R 7 Selected from H, C 1-4 Alkyl or C 1-4 alkyl halide;

[0045] R 8 and R 9 Independently selected from H, C 1-4 Alkyl or C 1-4 Halogenated alkyl.

[0046] Another aspect of the present invention provides a pharmaceutical composition comprising a preventively 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.

[0047] Another aspect of 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 in the preparation of a medicament for preventing or treating a disease or condition associated with PRMT5 activity.

[0048] Another aspect of 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, for use in preventing or treating a disease or condition associated with PRMT5 activity.

[0049] 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 or isotope-labeled compound thereof, or a pharmaceutical composition of the present invention.

[0050] Another aspect of the present invention provides methods of preparing the compounds of the present invention. DETAILED DESCRIPTION

[0051] definition

[0052] 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.

[0053] 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.").

[0054] 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 4, carbon atoms. For example, as used herein, the term "C 1-4 "Alkyl" refers to a linear or branched aliphatic hydrocarbon chain having 1 to 4 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl), which is optionally substituted with 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.).

[0055] As used herein, the term "heteroalkyl" refers to an optionally substituted alkyl group having one or more backbone chain atoms independently selected from atoms other than carbon, such as oxygen, nitrogen, sulfur, phosphorus, or combinations thereof. Numerical ranges (e.g., C 1-6 Assorted alkyl) refers to the number of carbons in the chain, which in this example includes 1-6 carbon atoms. For example, a -CH2OCH2CH3 group is referred to as a C3 assorted alkyl. Connection to the rest of the molecule can be through heteroatoms or carbon atoms in the assorted alkyl chain.

[0056] 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 1-4The 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.

[0057] 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 1-4 Hydroxyalkyl or C 1-3 Examples of hydroxyalkyl include, but are not limited to, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, -CH(OH)CH3, and the like.

[0058] As used herein, the term "alkoxy" refers to a group having an oxygen atom inserted into an alkyl group (as defined above) at any reasonable position, preferably a C 1-8 Alkoxy, C 1-6 Alkoxy, C 1-4 Alkoxy or C 1-3 Alkoxy. C 1-6 Representative examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, -CH2-OCH3, 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.

[0059] 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.

[0060] As used herein, the term "spiro" or "spirocyclyl" refers to a ring system formed by two or more ring structures sharing a ring atom. In the present invention, "spiro" can be "aliphatic spiro" or "spiroheterocycle". As used herein, the term "aliphatic spiro" refers to an all-carbon ring system formed by two or more rings sharing a carbon atom, which may contain one or more double bonds, including but not limited to 5-10 membered aliphatic spiro rings, such as 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 one ring atom, including but not limited to 5-10 membered spiroheterocycle, 6-10 membered spiroheterocycle, 6-10 membered nitrogen-containing spiroheterocycle, 6-10 membered oxygen-containing spiroheterocycle, 6-10 membered sulfur-containing spiroheterocycle, 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.

[0061] As used herein, the term "bridged ring" or "bridged cyclic group" refers to a ring system formed by two or more cyclic structures sharing two atoms that are not directly connected to each other. In the present invention, the "bridged ring" can be a "fat bridged ring" or a "bridged heterocycle". As used herein, the term "fat bridged ring" refers to an all-carbon ring system formed by two or more cyclic structures sharing two carbon atoms that are not directly connected to each other, which may contain one or more double bonds and can be divided into bicyclic, tricyclic, tetracyclic or polycyclic aliphatic bridged rings according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. "Fat bridged ring" includes but is not limited to 6-14 membered aliphatic bridged rings, 7-10 membered aliphatic bridged rings, for example 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.

[0062] As used herein, the term "cycloalkyl" refers to a saturated or unsaturated non-aromatic monocyclic or condensed polycyclic hydrocarbon ring group, including but not limited to monocyclic alkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, etc.) and condensed cycloalkyl. In the present invention, the cycloalkyl is optionally substituted by one or more (such as 1 to 3) identical or different substituents. The carbon atoms on the cycloalkyl are optionally substituted by 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 fused cycloalkyl, such as C 5-8 Fused cycloalkyl or C 5-6 Fused cycloalkyl.

[0063] 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.

[0064] As used herein, the term "heterocyclyl" or "heterocycle" refers to an aliphatic monocyclic or fused polycyclic 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 an oxo group (e.g., to form C=O, S(=O), or S(=O)2).

[0065] As used herein, the term "4-11 membered heterocyclyl" means a heterocyclyl group containing 4 to 11 ring atoms, including but not limited to a 4-10 membered heterocyclyl group, a 4-9 membered heterocyclyl group, a 4-8 membered heterocyclyl group, a 4-7 membered heterocyclyl group, a 5-6 membered heterocyclyl group, a 3-8 membered heterocyclyl group, a 3-7 membered heterocyclyl group, a 3-6 membered heterocyclyl group, a 4-7 membered nitrogen-containing heterocyclyl group, a 4-7 membered oxygen-containing heterocyclyl group, a 4-7 membered sulfur-containing heterocyclyl group, a 5-6 membered nitrogen-containing heterocyclyl group, a 5-6 membered oxygen-containing heterocyclyl group, a 5-6 membered sulfur-containing heterocyclyl group, and the like, wherein each of the "nitrogen-containing heterocyclyl group", "oxygen-containing heterocyclyl group" and "sulfur-containing heterocyclyl group" optionally further contains one or more other heteroatoms independently selected from oxygen, nitrogen and sulfur. Examples of 4-11 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.

[0066] The term "heterocyclyl" refers to -O-heterocyclyl, wherein heterocyclyl is as defined above. As used herein, "3-8 membered heterocyclyl" refers to a heterocyclyl having 3-8 (eg, 3, 4, 5, 6, 7 or 8) heteroatoms.

[0067] 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,

[0068] As used herein, the term "aryl" may contain 6 to 20 carbon atoms (C 6-20 Aryl), for example 6-14 carbon atoms (C 6-14 aryl) containing 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms; or 6-10 carbon atoms (C 6-10 The term "aryl" refers to a group consisting of 6, 7, 8, 9, or 10 carbon atoms. Examples include phenyl, naphthyl, indanyl, fluorenyl, and the like. "Aryl" includes a 6-membered aromatic carbocyclic ring, such as phenyl; a bicyclic ring containing at least one aromatic carbocyclic ring, such as naphthyl and indan; and a tricyclic ring containing at least one aromatic carbocyclic ring, such as fluorene. If an aryl substituent is bicyclic or tricyclic and at least one of the rings is non-aromatic, the linkage is considered to be through the aromatic ring. For example, an aryl group includes a 6-membered aromatic carbocyclic ring fused to a 5-7 membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, provided that the point of linkage is the aromatic carbocyclic ring.

[0069] As used herein, the term "heteroaryl" includes 5- to 10-membered (e.g., 5, 6, 7, 8, 9, or 10-membered) aromatic monocyclic rings containing 1 to 4 (e.g., 1, 2, 3, or 4) heteroatoms selected from N, O, and S, for example, 1 to 3 heteroatoms, i.e., 1, 2, or 3 heteroatoms, with the remainder being carbon atoms; 8- to 12-membered bicyclic rings containing 1 to 4 (e.g., 1, 2, 3, or 4) heteroatoms selected from N, O, and S, with the remainder being carbon atoms, wherein at least one heteroatom is present in the aromatic ring; and 11- to 14-membered tricyclic rings containing 1 to 4 (e.g., 1, 2, 3, or 4) heteroatoms selected from N, O, and S, with the remainder being carbon atoms, wherein at least one heteroatom is present in the aromatic ring.

[0070] As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

[0071] 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.

[0072] 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.

[0073] 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.

[0074] As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5 or 10, where reasonable.

[0075] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0076] 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.

[0077] 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.

[0078] 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:

[0079] 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.

[0080] 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).

[0081] 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.

[0082] 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).

[0083] 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.

[0084] 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.

[0085] 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.

[0086] 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.

[0087] The term "about" means within ±10%, preferably within ±5%, and more preferably within ±2% of the stated numerical value.

[0088] Compound

[0089] In some embodiments, the present invention provides a compound of Formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled compound thereof:

[0090] in:

[0091] R 1 for

[0092] R 2 and R 6 are independently selected from H, halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkoxy or 3-8 membered heterocyclic oxy;

[0093] R 3 、R 4 and R 5 are each independently selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution,

[0094] -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0095] -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0096] -XLC 6-10 Aryl, said aryl being optionally substituted by one or more halogen, CN, C1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,

[0097] -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,

[0098] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0099] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0100] or R 3 With R 4 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl,

[0101] or R 4 With R 5 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl,

[0102] The condition is that when R 1 for When R 3 、R 4 and R 5 At least one selected from the above -XLC 3-8 Cycloalkyl, -XL-3-8 membered heterocycloalkyl, -XLC 6-10 Aryl, -XL-5-10 membered heteroaryl, -XL-5-10 membered bridged ring group, -XL-5-10 membered spiro ring group, 3-8 membered heterocyclic group;

[0103] X is independently selected at each occurrence from O, S or NR 7 ;

[0104] L is independently a direct bond or -(CH2) n -;

[0105] n is 1 or 2;

[0106] R 7 Selected from H, C 1-4 Alkyl or C 1-4 alkyl halide;

[0107] R 8 and R 9 Independently selected from H, C 1-4 Alkyl or C 1-4 Halogenated alkyl.

[0108] In some embodiments, the present invention provides a compound of Formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, or isotopically labeled compound thereof:

[0109] in:

[0110] R 1 for

[0111] R 2 and R 6 are independently selected from H, halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 cycloalkoxy;

[0112] R 3 、R 4 and R 5 are each independently selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 substituted with a haloalkoxy group or a 3-6 membered heterocyclic group,

[0113] -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0114] -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0115] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0116] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0117] or R 3 With R 4 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl,

[0118] or R 4 With R 5 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl,

[0119] The condition is that when R 1 for When R 3 、R 4 and R 5 At least one selected from the above -XLC 3-8 Cycloalkyl, -XL-3-8 membered heterocycloalkyl, -XL-5-10 membered bridged ring group, -XL-5-10 membered spirocyclic group, 3-8 membered heterocyclic group;

[0120] X is independently selected at each occurrence from O, S or NR 7 ;

[0121] L is independently a direct bond or -(CH2) n -;

[0122] n is 1 or 2;

[0123] R 7 Selected from H, C 1-4 Alkyl or C 1-4 alkyl halide;

[0124] R 8 and R 9 Independently selected from H, C 1-4 Alkyl or C 1-4 Halogenated alkyl.

[0125] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 and R 6 are independently selected from H, halogen, CN, C 1-4 Alkyl, C 1-4 a haloalkyl group or a 3- to 8-membered heterocyclic group.

[0126] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 and R 6 are independently selected from H, halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl.

[0127] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 It is a 3-8 membered heterocyclic group.

[0128] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 for

[0129] In certain embodiments, the present invention provides compounds of formula I, wherein R 2 For CN.

[0130] In certain embodiments, the present invention provides compounds of formula I, wherein R 6 is F, Cl, Br or I.

[0131] In some embodiments, the compounds of the present invention have the structure shown in Formula IA:

[0132] in:

[0133] R 3 、R 4 and R 5 As defined above for Formula I.

[0134] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 3 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution,

[0135] -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0136] -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0137] -XLC 6-10 Aryl, said aryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,

[0138] -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,

[0139] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0140] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution.

[0141] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 3 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution,

[0142] -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0143] -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0144] -XLC 6-10 Aryl, said aryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,

[0145] -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,

[0146] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0147] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 haloalkyl substitution;

[0148] X is independently selected at each occurrence from O, S or NR 7 ;

[0149] L is independently selected at each occurrence from a direct bond or -(CH2) n -;

[0150] R 7 Selected from H, C 1-4 Alkyl or C 1-4 alkyl halide;

[0151] R 8 and R 9 Independently selected from H, C 1-4 Alkyl or C 1-4 Halogenated alkyl.

[0152] In other embodiments, the present invention provides compounds of formula I or formula IA, wherein R 3 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 substituted with a haloalkoxy group or a 3-6 membered heterocyclic group,

[0153] -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, C 1-4 Alkyl, C1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0154] -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0155] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0156] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution.

[0157] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 3 Selected from H, 3-6 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution,

[0158] -XLC 3-6 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0159] -XL-3-6 membered heterocyclic radicals, said heterocyclic radicals being optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0160] -XLC 6-10 Aryl, optionally substituted by one or more halogen, C 1-4 substituted by a haloalkyl group, a substituted or unsubstituted 5-10 membered heteroaryl group,

[0161] -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted with one or more substituted or unsubstituted 5-10 membered heteroaryl,

[0162] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0163] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution.

[0164] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 3 Selected from H, 3-6 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution,

[0165] -XLC 3-6 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0166] -XL-3-6 membered heterocyclic radicals, said heterocyclic radicals being optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0167] -XLC 6-10 Aryl, optionally substituted by one or more halogen, C 1-4substituted by a haloalkyl group, a substituted or unsubstituted 5-10 membered heteroaryl group,

[0168] -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted with one or more substituted or unsubstituted 5-10 membered heteroaryl,

[0169] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0170] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 haloalkyl substitution;

[0171] X is independently selected at each occurrence from O, S or NR 7 ;

[0172] L is independently selected at each occurrence from a direct bond or -(CH2) n -;

[0173] R 7 Selected from H, C 1-4 Alkyl or C 1-4 alkyl halide;

[0174] R 8 and R 9 Independently selected from H, C 1-4 Alkyl or C 1-4 Halogenated alkyl.

[0175] In other embodiments, the present invention provides compounds of formula I or formula IA, wherein R 3 Selected from 3-6 membered heterocyclic groups, wherein the heterocyclic groups are substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 substituted with a haloalkoxy group or a 3-6 membered heterocyclic group,

[0176] -XLC 3-6 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0177] -XL-3-6 membered heterocyclic radicals, said heterocyclic radicals being optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0178] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0179] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution.

[0180] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 3 Selected from H, 3-6 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution,

[0181] -XLC 3-6 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0182] -XL-3-6 membered heterocyclic radicals, said heterocyclic radicals being optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0183] -XLC 6-10 Aryl, optionally substituted by one or more halogen, C 1-4 substituted by a haloalkyl group, a substituted or unsubstituted 5-10 membered heteroaryl group,

[0184] -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted with one or more substituted or unsubstituted 5-10 membered heteroaryl,

[0185] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0186] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 haloalkyl substitution;

[0187] X is independently selected at each occurrence from O or NR 7 ;

[0188] L is independently selected at each occurrence from a direct bond or -(CH2)-;

[0189] R 7 Selected from H or C 1-4 alkyl;

[0190] R 8 and R 9 Independently selected from H, C 1-4 Alkyl or C 1-4 Halogenated alkyl.

[0191] In other embodiments, the present invention provides compounds of formula I or formula IA, wherein R 3 Selected from 3-6 membered heterocyclic groups, wherein the heterocyclic groups are substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 substituted with a haloalkoxy group or a 3-6 membered heterocyclic group,

[0192] -XLC 3-6 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0193] -XL-3-6 membered heterocyclic radicals, said heterocyclic radicals being optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0194] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0195] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 haloalkyl substitution;

[0196] X is independently selected at each occurrence from O or NR 7 ;

[0197] L is independently selected at each occurrence from a direct bond or -(CH2)-;

[0198] R 7 Selected from H or C 1-4 alkyl;

[0199] R 8 and R 9 Independently selected from H or C 1-4 alkyl.

[0200] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 3 Selected from H, azetidine, 2-azaspiro [3.3] heptane, wherein the azetidine is replaced by one or more F, CN, CF3, methyl, methoxy, ethoxy, CHF2 or N(CH3)2 substitution;

[0201] -XLC 3-6 Cycloalkyl, wherein the cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and the cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl is replaced by one or more F, hydroxyl, methyl, CHF2 or CF3 substitution;

[0202] -XL-3-6 membered heterocyclyl, wherein the heterocyclyl is selected from oxetane, azetidine, tetrahydrofuran, tetrahydropyran, piperidine or pyrrolidine, and the oxetane, azetidine, tetrahydrofuran, tetrahydropyran, piperidine, pyrrolidine is optionally substituted with one or more F, -C(O)CH3, methyl, ethyl, isopropyl or tert-butyl groups;

[0203] -XLC 6-10 Aryl, wherein the aryl is phenyl, wherein the phenyl is optionally substituted with one or more F, CF3, replace,

[0204] -XL-5-10 membered heteroaryl, said heteroaryl being pyridyl, said pyridyl being optionally substituted with one or more replace,

[0205] -XL-5-10 membered bridged ring group, wherein the bridged ring is bicyclo[1.1.1]pentane,

[0206] -XL-5-10 membered spirocyclyl, wherein the spirocyclyl is selected from 7-azaspiro[3.5]nonane, spiro[3.3]heptane or 2-oxaspiro[3.3]heptane, wherein the 7-azaspiro[3.5]nonane, spiro[3.3]heptane or 2-oxaspiro[3.3]heptane is optionally substituted with one or more methyl groups;

[0207] X at each occurrence is independently selected from O, NH or N(CH3);

[0208] L at each occurrence is independently selected from a direct bond or -(CH2)-.

[0209] In other embodiments, the present invention provides compounds of formula I or formula IA, wherein R 3 Selected from azetidine, wherein the azetidine is substituted by one or more F, CN, CF3, methyl or N(CH3)2;

[0210] -XLC 3-6 Cycloalkyl, wherein the cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and the cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl is substituted with one or more F, methyl or CF3;

[0211] -XL-3-6 membered heterocyclyl, wherein the heterocyclyl is selected from oxetane, azetidine, tetrahydrofuran, tetrahydropyran, piperidine or pyrrolidine, and the oxetane, azetidine, tetrahydrofuran, tetrahydropyran, piperidine, pyrrolidine is optionally substituted with one or more F, -C(O)CH3, methyl, ethyl, isopropyl or tert-butyl groups;

[0212] -XL-5-10 membered spirocyclyl, wherein the spirocyclyl is selected from 7-azaspiro[3.5]nonane, spiro[3.3]heptane or 2-oxaspiro[3.3]heptane, wherein the 7-azaspiro[3.5]nonane, spiro[3.3]heptane or -oxaspiro[3.3]heptane is optionally substituted with one or more methyl groups;

[0213] X at each occurrence is independently selected from O, NH or N(CH3);

[0214] L at each occurrence is independently selected from a direct bond or -(CH2)-.

[0215] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 3 Selected from

[0216] In other embodiments, the present invention provides compounds of formula I or formula IA, wherein R 3 Selected from

[0217] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 4 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 substituted with a haloalkoxy group or a 3-6 membered heterocyclic group,

[0218] -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0219] -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted,

[0220] -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution,

[0221] -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C1-4 Alkyl, C 1-4 Haloalkyl substitution.

[0222] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 4 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Substituted with a haloalkoxy group or a 3-6 membered heterocyclic group.

[0223] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 4 Selected from 3-8 membered heterocyclic groups, wherein the heterocyclic groups are substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Substituted with a haloalkoxy group or a 3-6 membered heterocyclic group.

[0224] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 4 Selected from H.

[0225] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 4 for

[0226] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 5 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Substituted with a haloalkoxy group or a 3-6 membered heterocyclic group.

[0227] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 5 Selected from H, C 1-4 Alkyl, halogen or 3-8 membered heterocyclic group, wherein the heterocyclic group is substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Substituted with a haloalkoxy group or a 3-6 membered heterocyclic group.

[0228] In other embodiments, the present invention provides compounds of formula I or formula IA, wherein R 5 is selected from halogen or 3-8 membered heterocyclic group, wherein the heterocyclic group is substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Substituted with a haloalkoxy group or a 3-6 membered heterocyclic group.

[0229] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 5 Selected from H, C 1-4 Alkyl or halogen.

[0230] In certain embodiments, the present invention provides compounds of Formula I or Formula IA, wherein R 5 is selected from H, methyl, F, Cl, Br or I, preferably H, methyl or Cl.

[0231] In other embodiments, the present invention provides compounds of formula I or formula IA, wherein R 5 is selected from F, Cl, Br or I, preferably Cl.

[0232] In certain embodiments, R 3 With R 4 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl.

[0233] In certain embodiments, R 4 With R 5 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl.

[0234] The present invention covers any combination of the above embodiments.

[0235] In some embodiments, compounds of the present invention include, but are not limited to:

[0236] In other embodiments, compounds of the present invention include, but are not limited to:

[0237] Preparation method

[0238] In certain embodiments, the present invention provides a method for preparing a compound of Formula IA comprising the steps of:

[0239] Route A

[0240] in:

[0241] PG is an amino protecting group, preferably tert-butyloxycarbonyl;

[0242] The remaining groups are as defined above;

[0243] The reaction conditions of each step are as follows:

[0244] Step 1: Compounds IA-1 and IA-2 undergo substitution reaction to generate compound IA-3;

[0245] The substitution reaction can be carried out in the presence of a base (eg, K2CO3, Cs2CO3, DIPEA, or Na2CO3) or NaH; a solvent such as THF, DMF, DMSO, or NMP can be used; and the reaction temperature is 0°C to 120°C.

[0246] Step 2: Compound IA-3 and IA-4 undergo Suzuki coupling reaction to generate compound IA-5;

[0247] The catalyst that can be used in the Suzuki coupling reaction is, for example, Pd(OAc)2, Pd(PPh3)4, Pd(dppf)Cl2, Pd(dppf)Cl2*DCM or cataCXiumAPdG3; the base that can be used is, for example, Cs2CO3, CsF, K3PO4, Na2CO3, KOAc, NaHCO3 or K2CO3; the solvent that can be used is, for example, 1,4-dioxane / H2O or DMF / H2O; and the reaction temperature is 40°C to 120°C.

[0248] Step 3: Compound IA-5 is deprotected under acidic conditions to generate a compound of formula IA;

[0249] The acid that can be used in the reaction is, for example, HCl or TFA; the solvent that can be used is, for example, DCM, THF, EA, 1,4-dioxane or methanol; and the reaction temperature is 20°C to 80°C.

[0250] Pharmaceutical compositions, preparations and methods of treatment

[0251] 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 isotopically labeled compound thereof and one or more pharmaceutically acceptable carriers.

[0252] In some embodiments, the present invention provides a pharmaceutical formulation, which is preferably a solid formulation, a semisolid formulation, a liquid formulation, or a gaseous formulation.

[0253] In some embodiments, the pharmaceutical composition or pharmaceutical formulation may further comprise one or more additional therapeutic agents.

[0254] In some embodiments, the pharmaceutical composition or pharmaceutical formulation is preferably administered orally, intravenously, intraarterially, subcutaneously, intraperitoneally, intramuscularly, or transdermally.

[0255] In some embodiments, the present invention provides the 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.

[0256] 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 the preparation of a medicament for modulating (e.g., reducing or inhibiting) PRMT5 activity.

[0257] 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.

[0258] In some embodiments, the present invention provides a method for preventing or treating a disease or condition associated with PRMT5 activity, comprising administering to an individual 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.

[0259] In some embodiments, the disease or condition associated with PRMT5 activity is preferably a cancer or tumor with MTAP deficiency.

[0260] In some embodiments, the cancer or tumor is preferably esophageal cancer, lung cancer, pancreatic cancer, glioblastoma, bile duct cancer, bladder cancer, mesothelioma, colorectal cancer, head and neck squamous cell carcinoma, lymphoma, etc.

[0261] 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.

[0262] 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).

[0263] The pharmaceutical compositions of the present invention can act systemically and / or locally. For this purpose, they can be administered by any suitable route.

[0264] For these administration routes, the pharmaceutical composition of the present invention can be administered in suitable dosage forms.

[0265] 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.

[0266] 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.

[0267] 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.

[0268] 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.

[0269] Unless otherwise indicated, as used herein, the term "treating" means reversing, alleviating, or eliminating the progression of the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. The term "preventing" includes inhibiting and delaying the onset of disease, and includes not only prevention prior to the development of a disease, but also prevention of recurrence of a disease after treatment.

[0270] 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.).

[0271] 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).

[0272] Example

[0273] 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.

[0274] The abbreviations used in this document have the following meanings:

[0275] 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). 1The reaction was confirmed by H NMR and / or MS. The reaction was monitored by TLC or LC-MS.

[0276] 1 H NMR spectroscopy was performed using a Bruker superconducting nuclear magnetic resonance spectrometer (model AVACE III HD 400 MHz).

[0277] LC / MS uses Aglient 1260 Infinity / Aglient 6120 Quadrupole.

[0278] TLC used silica gel GF 254 as the stationary phase.

[0279] Column chromatography generally uses 200-300 mesh silica gel (Qingdao Ocean) as the stationary phase.

[0280] Flash column chromatography was performed using a Biotage flash column chromatograph.

[0281] Prep-HPLC used Agilent 1260 and Waters 2489.

[0282] Microwave reactions were performed using a BiotageInitiator microwave reactor.

[0283] In the following examples, unless otherwise specified, the reaction temperature is room temperature (15-30°C).

[0284] The reagents used in this application were purchased from Acros Organics, Aldrich Chemical Company, or Teber Chemical Company.

[0285] Synthesis Example:

[0286] Intermediate Int-A: tert-butyl ((4-oxo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydrophthalazin-1-yl)methyl)carbamate

[0287] Step 1: Synthesis of 5-bromo-3-((dimethylamino)methylene)isobenzofuran-1(3H)-one (Int-Ab)

[0288] Int-Aa (46 g, 215.93 mmol), potassium tert-butoxide (2.42 g, 21.59 mmol), and DMF-DMA (165.71 g, 1.39 mol) were added to a flask and reacted at 110°C for 36 hours under nitrogen. After the reaction, the DMF-DMA was removed under reduced pressure, and the crude product was purified by silica gel column chromatography (mobile phase DCM:MeOH = 97:3) to obtain the intermediate Int-Ab (45.95 g). MS m / z (ESI): 268.1 [M+H] + .

[0289] Step 2: Synthesis of 6-bromo-4-((dimethylamino)methyl)phthalazin-1(2H)-one (Int-Ac)

[0290] Dissolve Int-Ab (12 g, 44.76 mmol) in ethanol, add hydrazine hydrate (4.69 g, 91.75 mmol), and place under nitrogen. Stir at room temperature for 30 minutes, then heat to 70°C and react for 12 hours. After the reaction, cool to room temperature, filter, rinse the filter cake with ethanol, and dry in vacuo to obtain the intermediate Int-Ac (8.50 g). MS m / z (ESI): 282.1 [M+H] + .

[0291] Step 3: Synthesis of 6-bromo-4-(chloromethyl)phthalazin-1(2H)-one (Int-Ad)

[0292] To anhydrous tetrahydrofuran (40 mL) was added Int-Ac (4 g, 14.18 mmol). Under nitrogen protection, isobutyl chloroformate (3.87 g, 28.36 mmol) was slowly added dropwise at 0°C. After addition, the temperature was naturally warmed to room temperature and the reaction was allowed to proceed for 16 hours. After completion of the reaction, the temperature was lowered to 0°C and 0.5 M dilute hydrochloric acid (100 mL) was slowly added dropwise with stirring, maintaining the reaction temperature below 10°C. After complete addition, the filter cake was rinsed with water and dried to obtain the intermediate Int-Ad (3.37 g). MS m / z (ESI): 273.2 [M+H] + .

[0293] Step 4: Synthesis of 2-(7-bromo-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)isoindoline-1,3-dione (Int-Ae)

[0294] Int-Ad (2 g, 7.31 mmol), potassium phthalimide (1.49 g, 8.04 mmol), and DMF (27 mL) were added to the reaction flask and allowed to react at room temperature for 16 hours. After completion, 3 mL of 6M hydrochloric acid was added, followed by 40 mL of water, and stirred for 15 minutes. The mixture was filtered, and the filter cake was rinsed with aqueous sodium bicarbonate solution and purified water, followed by drying to obtain the intermediate Int-Ae (2.76 g). MS m / z (ESI): 384.2 [M+H] + .

[0295] Step 5: Synthesis of 4-(aminomethyl)-6-bromophthalazin-1(2H)-one (Int-Af)

[0296] Int-Ae (2.76 g, 7.18 mmol), hydrazine hydrate (1.44 g, 28.74 mmol), and ethanol (30 mL) were added to the reaction flask and reacted at 80°C for 2 hours under nitrogen. After completion of the reaction, the mixture was cooled to room temperature, filtered, and the filter cake was rinsed with water and dried to obtain the intermediate Int-Af (1.67 g). MS m / z (ESI): 254.2 [M+H] + .

[0297] Step 6: Synthesis of tert-butyl ((7-bromo-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (Int-Ag)

[0298] Int-Af (1.67 g, 6.57 mmol) and triethylamine (2.0 g, 19.72 mmol) were added to dichloromethane (15 mL). A solution of (Boc)2O (12.87 g, 13.15 mmol) in dichloromethane (2 mL) was then added dropwise. The mixture was allowed to react at room temperature for 2 hours. After completion of the reaction, the mixture was filtered and the filter cake dried to obtain the intermediate Int-Ag (1.56 g). MS m / z (ESI): 354.1 [M+H] + .

[0299] Step 7: Synthesis of tert-butyl ((4-oxo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydrophthalazin-1-yl)methyl)carbamate (Int-A)

[0300] Int-Ag (747 mg, 2.11 mmol), Pd(dppf)Cl2 (154.32 mg, 210.90 μmol), pinacol diboronate (803.33 mg, 3.16 mmol), and KOAc (620.93 mg, 6.33 mmol) were added to 1,4-dioxane (8 mL) and reacted at 100°C for 2 hours under nitrogen. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (mobile phase: DCM:MeOH = 95:5) to afford the intermediate Int-A (1.10 g). MS m / z (ESI): 402.2 [M+H] + .

[0301] Intermediates Int-B and Int-C: 4-chloro-3,6-difluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)benzonitrile (Int-B) and 4-chloro-6-cyclopropyloxy-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)benzonitrile (Int-C)

[0302] Step 1: Synthesis of 4-chloro-3,6-difluoro-2-iodobenzonitrile (Int-Bb)

[0303] To a reaction flask, compound Int-Ba (5.00 g, 28.81 mmol), Pd(OAc)2 (646.83 mg, 2.88 mmol), NIS (12.96 g, 57.62 mmol), p-TsOH (2.48 g, 14.41 mmol), and DCE (100 mL) were added and reacted at 75°C for 16 hours under nitrogen. After completion, the reaction was cooled to room temperature, diluted with DCM, and washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase: PE:EA = 95:5) to obtain compound Int-Bb (4.94 g).

[0304] Step 2: Synthesis of 4-chloro-3,6-difluoro-2-(1-methyl-1H-pyrazol-5-yl)benzonitrile (Int-Bc)

[0305] To a reaction flask, compound Int-Bb (3.00 g, 10.02 mmol), Pd(dtbpf)Cl2 (652.96 mg, 1.00 mmol), 1-methyl-1H-pyrazole-5-boronic acid pinacol ester (5.21 g, 25.05 mmol), Na2CO3 (2.12 g, 20.04 mmol), 1,4-dioxane (20 mL), and H2O (4 mL) were added. The mixture was reacted at 75°C under nitrogen for 16 hours. After completion, the reaction was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (mobile phase: PE:EA = 78:22) to obtain compound Int-Bc (733 mg). MS m / z (ESI): 254.2 [M+H] + .

[0306] Step 3: Synthesis of 4-chloro-3,6-difluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)benzonitrile (Int-B)

[0307] Compound Int-Bc (733 mg, 2.89 mmol), NIS (1.30 g, 5.78 mmol), and AcOH (20 mL) were added to the reaction flask and reacted at 80°C for 3 hours under nitrogen. After completion of the reaction, the acetic acid was removed by concentration under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase PE:EA = 85:15) to obtain compound Int-B (810 mg). MS m / z (ESI): 380.1 [M+H] + .

[0308] Step 4: Synthesis of 4-chloro-6-cyclopropyloxy-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)benzonitrile (Int-C)

[0309] Under ice, NaH (25.3 mg, 632.4 μmol) was added to a solution of cyclopropanol (30.6 mg, 527.0 μmol) in THF (10.0 mL). After stirring for 10 minutes, Int-B (200.0 mg, 527.0 μmol) was added and the mixture was allowed to react at 50°C for 1.5 hours. After completion, the reaction was quenched with water and concentrated under reduced pressure to remove the solvent. The crude product was purified by silica gel column chromatography (mobile phase: PE:EA = 90:10) to obtain compound Int-C (200 mg). MS m / z (ESI): 417.9 [M+H] + .

[0310] Example 1: 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(oxetan-3-ylmethoxy)benzonitrile (1)

[0311] Step 1: Synthesis of 4-chloro-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)-6-(oxetane-3-ylmethoxy)benzonitrile (1b)

[0312] Compound Int-B (50 mg, 131.74 μmol), 1a (58.00 mg, 658 μmol), K2CO3 (54.62 mg, 395.22 μmol), and DMF (2 mL) were added to a reaction flask and reacted at 75°C for 18 hours. After completion, the mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate and washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase PE:EA = 68:32) to obtain compound 1b (26 mg). MS m / z (ESI): 448.1 [M+H] + .

[0313] Step 2: Synthesis of tert-butyl ((7-(5-(3-chloro-6-cyano-2-fluoro-5-(oxetan-3-ylmethoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (1c)

[0314] Compound 1b (26 mg, 50.08 μmol), Int-A (25.64 mg, 63.89 μmol), Pd(dppf)Cl2 (4.71 mg, 5.81 μmol), and NaHCO3 (11.71 mg, 139.40 μmol) were added to 1,4-dioxane (5 mL) and H2O (0.5 mL). The mixture was reacted at 80°C for 4 hours under nitrogen. After completion, the reaction was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH = 90:10) to obtain compound 1c (25 mg). MS m / z (ESI): 595.1 [M+H] + .

[0315] Step 3: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(oxetane-3-ylmethoxy)benzonitrile (1)

[0316] Compound 1c (25 mg, 42.02 μmol) was added to dichloromethane (8 mL). TFA (383.24 mg, 3.36 mmol) was added dropwise under an ice bath. The mixture was allowed to warm to room temperature and allowed to react for 2 hours. After completion of the reaction, triethylamine was added to adjust the mixture to a weak base. The mixture was concentrated under reduced pressure, and the crude product was purified by Pre-HPLC to yield compound 1 (2.20 mg). MS m / z (ESI): 495.1 [M+H] + .

[0317] 1 H NMR (400MHz, DMSO-d6) δ12.55(s,1H),8.30(s,1H),8.15(d,J=8.4Hz,1H),7.95(d,J=6.0Hz,1H),7.80(d,J=1.2Hz,1H),7.64(dd,J=8.4, 1.6Hz,1H),4.69(dd,J=8.0,6.4Hz,2H),4.48(d,J=6.4Hz,2H),4.42(td,J=6.4,2.0Hz,2H),3.92(s,2H),3.78(s,3H),3.48–3.37(m,2H).

[0318] The following compounds were prepared by the method and general steps described in Example 1. 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.

[0319] Example 9: 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(tetrahydro-2H-pyran-4-yl)oxy)benzonitrile (9)

[0320] Step 1: Synthesis of 4-chloro-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)-6-((tetrahydro-2H-pyran-4-yl)oxy)benzonitrile (9b)

[0321] Compound 9a (26.91 mg, 263.48 μmol) and THF (5 mL) were added to the reaction flask under an ice bath. NaH (12.65 mg, 316.18 μmol) was then slowly added and stirred for 10 min. Finally, a solution of Int-B (100 mg, 263.48 μmol) in THF (1 mL) was added dropwise. The mixture was reacted at 50°C for 1 hr. After completion of the reaction, the mixture was cooled to room temperature, quenched with a small amount of methanol, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase: DCM:MeOH = 97:3) to obtain compound 9b (77.0 mg). MS m / z (ESI): 462.0 [M+H] + .

[0322] Step 2: Synthesis of tert-butyl ((7-(5-(3-chloro-6-cyano-2-fluoro-5-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (9c)

[0323] Compound 9b (77.0 mg, 166.79 μmol), Int-A (73.62 mg, 183.47 μmol), Pd(dppf)Cl2 (13.52 mg, 16.68 μmol), and NaHCO3 (42.03 mg, 500.37 μmol) were added to 1,4-dioxane (5 mL) and H2O (0.5 mL). The mixture was reacted at 80°C for 5 hours under nitrogen. After completion, the reaction was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase: PE:EA = 0:100) to obtain compound 9c (20.0 mg). MS m / z (ESI): 609.1 [M+H] + .

[0324] Step 3: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(tetrahydro-2H-pyran-4-yl)oxy)benzonitrile (9)

[0325] Compound 9c (20.0 mg, 32.84 μmol) was added to dichloromethane (10 mL). TFA (299.54 mg, 2.63 mmol) was added dropwise under an ice bath. The mixture was allowed to warm to room temperature and allowed to react for 2 hours. After completion of the reaction, triethylamine was added to adjust the mixture to a weak base. The mixture was concentrated under reduced pressure, and the crude product was purified by Pre-HPLC to yield compound 9 (4.82 mg). MS m / z (ESI): 509.1 [M+H] + .

[0326] 1 H NMR (400MHz, DMSO-d6) δ12.59(s,1H),8.30(s,1H),8.15(d,J=8.0Hz,1H),8.04(d,J=6.0Hz,1H),7.78(d,J=1.2Hz,1H),7.63(dd ,J=8.4,1.6Hz,1H),5.01–4.83(m,1H),3.95(s,2H),3.87–3.71(m,5H),3.57–3.43(m,2H),2.08–1.92(m,2H),1.68–1.57(m,2H).

[0327] The following compounds were prepared by the method and general steps described in Example 9. 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.

[0328] Example 15: 2-(4-(4-(Aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-difluorocyclobutyloxy)-3-fluorobenzonitrile (15 / 16)

[0329] Step 1: Synthesis of 4-chloro-6-(3,3-difluorocyclobutyloxy)-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)benzonitrile (15b)

[0330] Compound Int-B (100 mg, 263 μmol), 15a (42.7 mg, 395 μmol), and K2CO3 (91.0 mg, 658 μmol) were added to DMF (1 mL) and heated to 80°C for 16 hours. After completion of the reaction, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phase was concentrated, and the crude product was purified by Prep-TLC (PE / EA = 2 / 1) to obtain compound 15b (50 mg). MS m / z (ESI): 467.9 [M+H] + .

[0331] Step 2: Synthesis of tert-butyl ((7-(5-(3-chloro-6-cyano-5-(3,3-difluorocyclobutyloxy)-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (15c)

[0332] Compound 15b (50.0 mg, 106 μmol), Int-A (60.0 mg, 149 μmol), CsF (97.4 mg, 641 μmol), and cataCXiumAPdG3 (7.79 mg, 10.6 μmol) were added to 1,4-dioxane (1 mL) and H2O (0.2 mL) and reacted at 60°C for 12 hours under nitrogen. After completion, the reaction was cooled to room temperature, quenched with water, and extracted with ethyl acetate. The mixture was washed with saturated brine and dried over anhydrous sodium sulfate. The mixture was filtered and concentrated under reduced pressure. The crude product was purified by Prep-TLC (developing solvent: EA:PE = 1:1) to obtain compound 15c (40 mg). Compound 15c was resolved by SFC (column: REGIS (S,S) WHELK-O1 (250 mm x 25 mm, 10 μm); mobile phase: 25% [0.1% NH 3* H2O in MeOH solution] + 75% liquid CO2) to give compound 15c peak 1 (20 mg), MS m / z (ESI): 615.3 [M+H] + and compound 15c peak 2 (20 mg), MS m / z (ESI): 615.3 [M+H] + .

[0333] Step 3: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-difluorocyclobutyloxy)-3-fluorobenzonitrile (15 / 16)

[0334] Compound 15c peak 1 (20.0 mg, 32.5 μmol) was added to a HCl solution in EA (2 mL) and allowed to react at room temperature for 1 hour. After the reaction, the reaction solution was concentrated to dryness to obtain compound 15 (14.7 mg). MS m / z (ESI): 515.2 [M+H] + .

[0335] 1 H NMR: (400MHz DMSO-d6) δ12.89(s,1H),8.37(s,1H),8.14(d,J=8.4Hz,1H),7.88(s,1H),7.77(d,J=6.0Hz,1H),7.46(dd,J=1.2,8.4Hz,1H),5.05(br dd,J=4.0,7.6Hz,1H),4.39(s,2H),3.79(s,3H),3.29(br d,J=3.2Hz,2H),2.94-2.72(m,2H).

[0336] Compound 15c peak 2 (20.0 mg, 32.5 μmol) was added to a HCl solution in EA (2 mL) and allowed to react at room temperature for 1 hour. After the reaction, the reaction solution was concentrated to dryness to obtain compound 16 (14.9 mg). MS m / z (ESI): 515.1 [M+H] + .

[0337] 1 H NMR: (400MHz DMSO-d6) δ12.88(s,1H),8.37(s,1H),8.15(d,J=8.4Hz,1H),7.88(s,1H),7.78(d,J=5.6Hz,1H),7.48(br d,J=9.2Hz,1H),5.10-4.99(m,1H),4.37(s,2H),3.80(s,3H),3.28-3.16(m,2H),2.90-2.76(m,2H).

[0338] Example 17: 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-dimethylcyclobutyl)methoxy)-3-fluorobenzonitrile (17)

[0339] Step 1: Synthesis of 4-chloro-6-((3,3-dimethylcyclobutyl)methoxy)-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)benzonitrile (17b)

[0340] Compound 17a (21.7 mg, 190 μmol) and THF (5 mL) were added to the reaction flask under an ice bath. NaH (9.5 mg, 237 μmol) was then slowly added and stirred for 30 min. Finally, a solution of Int-B (60.0 mg, 158 μmol) in THF (1 mL) was added dropwise. The mixture was allowed to react at 25°C for 2 hr. After completion, the reaction was cooled to room temperature, quenched with a small amount of water, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase: PE:EA = 5:1) to obtain compound 17b (50.0 mg). MS m / z (ESI): 473.9 [M+H] + .

[0341] Step 2: Synthesis of tert-butyl ((7-(5-(3-chloro-6-cyano-2-fluoro-5-(3,3-dimethylcyclobutyl)methoxy)phenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (17c)

[0342] Compound 17b (38.5 mg, 81.3 μmol), Int-A (42.4 mg, 106 μmol), CsF (74.1 mg, 488 μmol), and cataCXium A Pd-G3 (5.9 mg, 8.13 μmol) were added to 1,4-dioxane (10 mL) and H2O (2.5 mL). The mixture was reacted at 60°C for 12 hours under nitrogen. After completion, the reaction was cooled to room temperature and quenched with water. The mixture was then extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by prep-HPLC (mobile phase: H2O(NH4HCO3) / ACN = 1:1) to afford compound 17c (33.0 mg). MS m / z (ESI): 621.6 [M+H] + .

[0343] Step 3: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-dimethylcyclobutyl)methoxy)-3-fluorobenzonitrile (17)

[0344] Compound 17c (30.0 mg, 48.3 μmol) was added to HCl / EtOAc (4 M, 5.00 mL) under an ice bath and allowed to warm to room temperature for 1 hour. After completion, the reaction was quenched with water and extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by Pre-HPLC to yield compound 17 (7.00 mg). MS m / z (ESI): 521.0 [M+H] + .

[0345] 1 H NMR (400MHz, DMSO-d6) δ12.89(s,1H),8.46-8.39(m,2H),8.38-8.34(m,1H),8.14(d,J=8.4Hz,1H),7.92-7.85(m,2H),7.46-7.40(m,1H),4.4 4-4.35(m,2H),4.23-4.15(m,2H),3.83-3.75(m,3H),2.04-1.96(m,1H ),1.82-1.76(m,2H),1.71(d,J=6.4Hz,2H),1.14(s,3H),1.04(s,3H).

[0346] The following compounds were prepared by the method and general steps described in Example 17. 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.

[0347] Example 21: 2-(4-(4-(Aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluorobenzonitrile (21 and 60, 61)

[0348] Step 1: Synthesis of 4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)benzonitrile (21b)

[0349] Compound Int-B (50 mg, 131.74 μmol), 21a (51 mg, 395.22 μmol), and DIPEA (85 mg, 658.71 μmol) were added to NMP (1.5 mL) and microwave-heated at 100°C for 8 hours. After completion, the reaction was cooled to room temperature, quenched with water, and extracted with ethyl acetate. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase EA:PE = 15:85) to obtain compound 21b (35 mg).

[0350] Step 2: Synthesis of tert-butyl (7-(5-(3-chloro-6-cyano-5-(3,3-difluoroazetidin-1-yl)-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (21c)

[0351] Compound 21b (35 mg, 77.33 μmol), Int-A (34 mg, 85.06 μmol), Pd(dppf)Cl2 (6 mg, 7.73 μmol), and NaHCO3 (19 mg, 231.99 μmol) were added to 1,4-dioxane (3 mL) and H2O (0.5 mL). The mixture was reacted at 80°C for 5 hours under nitrogen. After completion, the reaction was cooled to room temperature, quenched with water, and extracted with ethyl acetate. The mixture was washed with saturated brine and dried over anhydrous sodium sulfate. The mixture was filtered and concentrated under reduced pressure. The crude product was purified by Prep-TLC (developing solvent EA:PE = 1:1) to obtain compound 21c (20 mg). MS m / z (ESI): 601.3 [M+H] + .

[0352] Step 3: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluorobenzonitrile (21)

[0353] Compound 21c (25 mg, 38.12 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of TFA (0.5 mL). The mixture was reacted at room temperature under nitrogen for 2 hours. After completion of the reaction, the solvent was removed by concentration under reduced pressure, and the mixture was dissolved in DCM and adjusted to a weak alkalinity by adding triethylamine. The reaction was then concentrated under reduced pressure. The crude product was purified by Pre-HPLC to yield compound 21 (19.05 mg). MS m / z (ESI): 500.1 [M+H] + .

[0354] 1 H NMR (400MHz, DMSO-d6) δ12.53(br,1H),8.32–8.29(m,1H),8.16(d,J=8.4Hz,1H),7.83(d,J=1.6Hz,1 H),7.67(dd,J=8.4,1.6Hz,1H),7.31(d,J=6.0Hz,1H),4.71–4.55(m,4H),3.92(s,2H),3.78(s,3H).

[0355] Step 4: Separation of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluorobenzonitrile (21) stereoisomers Compound 60 and Compound 61

[0356] Compound 21 was separated by prep-SFC (chiral column: DAICEL CHIRALPAK IC (250 mm*30 mm, 10 μm); mobile phase: [CO2-EtOH (0.05% DEA)]) to give compound 60 (8.78 mg, retention time = 1.073 min), MS m / z (ESI): 500.1 [M+H] + and 61 (13.48 mg, retention time = 1.730 min), MS m / z (ESI): 500.1 [M+H] + .

[0357] Compound 60: 1H NMR (400MHz, DMSO-d6) δ12.47(s,1H),8.29(s,1H),8.17(d,J=8.2Hz,1H),7.82(d,J=1.0Hz,1H),7.70( dd,J=8.4,1.4Hz,1H),7.31(d,J=6.3Hz,1H),4.63(dd,J=25.0,12.3Hz,4H),3.86(s,2H),3.79(s,3H).

[0358] Compound 61: 1 H NMR (400MHz, DMSO-d6) δ12.45(s,1H),8.29(s,1H),8.17(d,J=8.2Hz,1H),7.82(d,J=1.5Hz,1H),7.71( dd,J=8.3,1.5Hz,1H),7.31(d,J=6.2Hz,1H),4.63(dd,J=25.9,11.7Hz,4H),3.83(s,2H),3.79(s,3H).

[0359] The following compounds were prepared by the method and general steps described in Example 21. 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.

[0360] Example 42: 2-(4-(4-(Aminomethyl)-1-oxo-1,2-dihydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (42)

[0361] Step 1: Synthesis of methyl 4-bromo-2-(bromomethyl)benzoate (42b)

[0362] Compound 42a (10.0 g, 43.6 mmol) was dissolved in CCl4 (100 mL), followed by the addition of BPO (528 mg, 2.18 mmol) and NBS (9.32 g, 52.4 mmol). The mixture was heated to 80°C under a nitrogen atmosphere for 12 hours. After completion, the reaction was cooled to room temperature, quenched with water, and extracted with dichloromethane. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by HPLC (mobile phase: 0.1% FA) to afford compound 42b (9.8 g).

[0363] Step 2: Synthesis of ethyl 4-bromo-2-(cyanomethyl)benzoate (42c)

[0364] A solution of NaCN (910 mg, 18.5 mmol) in water (10 mL) was added to a solution of compound 42b (5.0 g, 16.2 mmol) in ethanol (50 mL) at 0°C. The mixture was reacted at 80°C for 2 hours under nitrogen. After completion, the reaction was cooled to room temperature, quenched with water, and extracted with ethyl acetate. The mixture was washed with saturated brine and dried over anhydrous sodium sulfate. The mixture was filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (developing solvent: PE:EA = 0:1) to obtain compound 42c (1.2 g).

[0365] Step 3: Synthesis of 6-bromo-1-oxo-1,2-dihydroisoquinoline-4-carbonitrile (42d)

[0366] Compound 42c (1.2 g, 4.48 mmol) and 1,3,5-triazine (435 mg, 5.37 mmol) were dissolved in MeOH (20 mL). A solution of NaOMe (1.21 g, 223 mmol) in MeOH (5 mL) was added and the mixture was reacted at 25°C for 20 min under nitrogen. After completion, the reaction was quenched by the addition of aqueous NH4Cl (50 mL). The mixture was diluted with ethyl acetate and stirred at 25°C for 20 min. The filter cake was collected by filtration to obtain compound 42d (900.0 mg).

[0367] Step 4: Synthesis of 1-oxo-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2-dihydroisoquinoline-4-carbonitrile (42e)

[0368] Compound 42d (200 mg, 80.3 μmol), B2Pin2 (305 mg, 1.20 mmol), Pd(dppf)Cl2 (58.7 mg, 80.3 μmol), and KOAc (157 mg, 1.61 mmol) were dissolved in 1,4-dioxane (3 mL) and reacted at 80°C for 12 hours under nitrogen. After completion of the reaction, the mixture was concentrated and purified by silica gel column chromatography (developing solvent: PE:EA = 1:1) to obtain compound 42e (180.0 mg). MS m / z (ESI): 214.5 [M-81]. + .

[0369] Step 5: Synthesis of 4-(aminomethyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinolin-1(2H)-one (42f)

[0370] Compound 42e (50.0 mg, 168 μmol) was added to a solution of Raney-Ni (14.4 mg, 168 μmol) in NH3 / MeOH (1 mL). The mixture was reacted at 20°C under a H2 atmosphere (15 psi) for 12 hours. After completion of the reaction, the mixture was directly filtered to obtain compound 42f (25.0 mg). MS m / z (ESI): 219.0 [M-81]. + .

[0371] Step 6: Synthesis of tert-butyl ((1-oxo-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2-dihydroisoquinolin-4-yl)methyl)carbamate (42 g)

[0372] Compound 42f (25.0 mg, 83 μmol) was dissolved in CH2Cl2 (0.5 mL), and Boc2O (36.3 mg, 166 μmol) and TEA (25.2 mg, 249 μmol) were added. After nitrogen displacement, the reaction was incubated at 25°C for 10 min. The mixture was concentrated and washed with petroleum ether (2 mL) to afford compound 42g (15.0 mg). MS m / z (ESI): 401.0 [M+H] + .

[0373] Step 7: Synthesis of tert-butyl ((6-(5-(3-chloro-6-cyano-5-cyclopropyloxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-1-oxo-1,2-dihydroisoquinolin-4-yl)methyl)carbamate (42h)

[0374] Compound 42g (15.0 mg, 37.4 μmol), Int-C (15.6 mg, 37.4 μmol), CsF (34.1 mg, 224 μmol), and cataCXium A Pd-G3 (2.7 mg, 3.7 μmol) were added to 1,4-dioxane (0.5 mL) and H2O (0.1 mL). The mixture was reacted at 60°C for 12 hours under nitrogen. After completion, the reaction was cooled to room temperature and quenched with water. The mixture was then extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by prep-HPLC (mobile phase: PE / EA = 1:1) to yield compound 42h (10.0 mg). MS m / z (ESI): 564.2 [M+H] + .

[0375] Step 8: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (42)

[0376] Compound 42h (10.0 mg, 48.3 μmol) was added to HCl / EtOAc (4 M, 5.00 mL) under ice-cooling, and then heated to 25°C for 30 min. After completion, the reaction was quenched with water and extracted three times with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by Pre-HPLC ([water(HCl)-ACN]; B%: 12%-42%) to yield compound 42 (2.47 mg). MS m / z (ESI): 464.2 [M+H] + .

[0377] 1 H NMR(400MHz,DMSO-d6)δ11.47(d,J=6.0Hz,1H),8.39(s,1H),8.26(br s,2H),8.07(d,J=8.4Hz,1H),8.01(d,J=6.0Hz,1H),7.82(d,J=1.2Hz,1H),7.40(d,J=6.0Hz,1H),7.03(dd, J=1.2,8.4Hz,1H),4.25-4.18(m,1H),4.17-4.04(m,2H),3.76(s,3H),0.96-0.87(m,2H),0.87-0.77(m,2H)

[0378] Example 43: 2-(4-(4-(Aminomethyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (43)

[0379] Step 1: Synthesis of tert-butyl ((1-oxo-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinolin-4-yl)methyl)carbamate (43b)

[0380] Compound 42g (10.0 mg, 24.9 μmol) was dissolved in EtOH (2 mL), and Pd / C (0.1 g, 10% purity) was added. The mixture was reacted at 50°C under H2 (50 psi) for 12 hours. The mixture was directly filtered to obtain compound 43b (15.0 mg). MS m / z (ESI): 347.0 [M-55]. + .

[0381] Step 2: Synthesis of tert-butyl ((6-(5-(3-chloro-6-cyano-5-cyclopropyloxy-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-4-yl)methyl)carbamate (43c)

[0382] Compound 43b (15.0 mg, 37.3 μmol), Int-C (15.5 mg, 37.3 μmol), Pd(dppf)Cl2 (2.7 mg, 3.7 μmol), and KOAc (10.9 mg, 111 μmol) were added to 1,4-dioxane (0.5 mL) and H2O (0.5 mL). The mixture was reacted at 80°C for 8 hours under nitrogen. After completion of the reaction, the mixture was cooled to room temperature and concentrated. The crude product was purified by prep-TLC (mobile phase PE / EA = 1:2) to obtain compound 43c (10.0 mg). MS m / z (ESI): 510.1 [M-55]. + .

[0383] Step 3: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (43)

[0384] Compound 43c (10.0 mg, 17.6 μmol) was added to HCl / EtOAc (4 M, 2.0 mL) under ice-cooling, and then the temperature was raised to 25°C for 1 hour. The mixture was concentrated under reduced pressure to obtain compound 43 (9.33 mg). MS m / z (ESI): 466.0 [M+H] + .

[0385] 1 H NMR (400MHz, DMSO-d6) δ8.10-8.00 (m, 3H), 7.98 (dd, J = 2.4, 5.6Hz, 1H), 7.89-7. 82(m,1H),7.75(d,J=8.0Hz,1H),7.45-7.36(m,1H),7.04-6.96(m,1H),4.19(br s,1H),3.75(d,J=2.8Hz,3H),3.60-3.51(m,1H),3.47-3.41(m,1H),3.2 4-3.17(m,1H),3.09-2.99(m,1H),2.97-2.82(m,1H),0.94-0.78(m,4H).

[0386] Example 50: 2-(4-(4-(Aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile (50 / 51)

[0387] Step 1: Synthesis of 4-chloro-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)-6-(1-methylcyclopropyloxy)benzonitrile (20b)

[0388] Compound 20a (68.3 mg, 948 μmol) and THF (3 mL) were added to the reaction flask under an ice bath. NaH (47.4 mg, 1.19 mmol) was then slowly added and stirred for 30 min. Finally, a solution of Int-B (300 mg, 790 μmol) in THF (1 mL) was added dropwise. The mixture was reacted at 50°C for 2 hr. After completion of the reaction, the temperature was lowered to 0°C and quenched with 5 mL of water. The mixture was then extracted with ethyl acetate. The organic phases were combined and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase: PE:EA = 3:1) to obtain compound 20b (200.0 mg). MS m / z (ESI): 431.8 [M+H] + .

[0389] Step 2: Synthesis of tert-butyl ((7-(5-(3-chloro-6-cyano-2-fluoro-5-(1-methylcyclopropyloxy)phenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (20c)

[0390] Compound 20b (200.0 mg, 463 μmol), Int-A (223.0 mg, 556 μmol), CsF (211.0 mg, 1.39 mmol), and cataCXiumAPdG3 (33.8 mg, 46.3 μmol) were added to 1,4-dioxane (3 mL) and H2O (0.6 mL). The mixture was reacted at 60°C under nitrogen for 12 hours. After completion, the reaction was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by Prep-TLC (developing solvent: EA:PE = 1:1) to obtain compound 20c. Compound 20c was separated by SFC (column: DAICEL CHIRALPAK IC (250 mm*30 mm, 10 μm); mobile phase: [Neu-ETOH]; B%: 40%-40%, A 4.4; 60 min) to afford compound 20c peak 1 (45.0 mg), MS m / z (ESI): 579.1 [M+H] + and compound 20c peak 2 (40.0 mg), MS m / z (ESI): 579.1 [M+H] + .

[0391] Step 3: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile (50 / 51)

[0392] Compound 20c peak 1 (45.0 mg, 77.7 μmol) was added to a 3 mL HCl solution in EA and allowed to react at 25°C for 0.5 hr. After completion of the reaction, the reaction solution was concentrated to dryness and analyzed by pre-HPLC (column: Welch Xtimate C18 150*25mm*5um; mobile phase: [water(HCl)-ACN]; B%: 12%-42%, 8 min) to yield compound 50 (29.8 mg). MS m / z (ESI): 479.1 [M+H] + .

[0393] 1 H NMR: (400MHz DMSO-d6)δ12.89(s,1H),8.52(br s,3H),8.37(s,1H),8.14(d,J=8.4Hz,1H),7.88(d,J=1.2Hz,1H),7.81(d,J=6.0Hz,1H),7.44(dd,J= 1.6,8.4Hz,1H),4.50-4.27(m,2H),3.78(s,3H),1.59(s,3H),1.09-0.98(m,2H),0.95-0.85(m,2H).

[0394] Compound 20c, peak 2 (40.0 mg, 69.1 μmol), was added to a 2 mL solution of HCl in EA and allowed to react at 25°C for 0.5 hr. After completion of the reaction, the reaction solution was concentrated to dryness and analyzed by pre-HPLC (column: Welch Xtimate C18 150*25mm*5um; mobile phase: [water(HCl)-ACN]; B%: 11%-41%, 8 min) to afford compound 51 (24.0 mg). MS m / z (ESI): 479.1 [M+H] + .

[0395] 1H NMR: (400MHz DMSO-d6)δ12.89(s,1H),8.51(br s,3H),8.37(s,1H),8.14(d,J=8.4Hz,1H),7.88(s,1H),7.81(d,J=6.0Hz,1H),7.47-7.41( m,1H),4.47-4.31(m,2H),3.78(s,3H),1.59(s,3H),1.10-0.96(m,2H),0.95-0.85(m,2H).

[0396] Example 52: 2-(4-(4-(Aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile (52)

[0397] Step 1: Synthesis of 4-chloro-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)-6-(3-methylazetidin-1-yl)benzonitrile (52b)

[0398] Compound 52a (17.0 mg, 158 μmol) and K2CO3 (65.6 mg, 474 μmol) were added to DMF (1.0 mL) and stirred at 25°C for 10 minutes. Int-B (60.0 mg, 158 μmol) was then added and the reaction was heated to 80°C and stirred for 12 hours. After completion, the reaction was cooled to room temperature, quenched with water, and extracted with ethyl acetate. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase: PE:EA = 3:1) to obtain compound 52b (40.0 mg). MS m / z (ESI): 430.9 [M+H] +

[0399] Step 2: Synthesis of tert-butyl ((7-(5-(3-chloro-6-cyano-2-fluoro-5-(3-methylazetidin-1-yl)phenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)formate (52c)

[0400] Compound 52b (40.0 mg, 92.9 μmol), Int-A (52.2 mg, 130 μmol), CsF (42.3 mg, 278 μmol), and cataCXiumAPdG3 (6.7 mg, 9.3 μmol) were added to 1,4-dioxane (1.0 mL) and H2O (0.2 mL). The mixture was reacted at 60°C for 12 hours under nitrogen. After completion, the reaction was cooled to room temperature, filtered, and concentrated under reduced pressure. The crude product was purified by Prep-TLC (developing solvent EA:PE = 1:1) to obtain compound 52c (20.0 mg). MS m / z (ESI): 578.2 [M+H] + .

[0401] Step 3: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile (52)

[0402] Compound 52c (20.0 mg, 34.6 μmol) was added to dichloromethane (2.0 mL). The mixture was cooled to 0°C and TFA (78.9 mg, 692.0 μmol, 0.05 mL) was added dropwise. The mixture was then reacted at 25°C for 1 hour. After completion of the reaction, the solvent was removed by concentration under reduced pressure. The crude product was purified by Pre-HPLC (column: Phenomenex C18 150*25mm*10um; mobile phase: [water (NH4HCO3)-ACN]; B%: 24%-54%, 8 min) to afford compound 52 (7.95 mg). MS m / z (ESI): 478.1 [M+H] + .

[0403] 1 H NMR (400MHz, DMSO-d6) δ12.49(br s,1H),8.27(s,1H),8.17(d,J=8.4Hz,1H),7.79(d,J= 1.5Hz,1H),7.70(dd,J=1.6,8.3Hz,1H),7.04(d,J=6.3Hz,1H),4.33-4.17(m,2H),3.87(s ,2H),3.76(s,3H),3.75-3.65(m,2H),2.77(dt,J=7.0,13.6Hz,1H),1.22(d,J=6.9Hz,3H).

[0404] The following compounds were prepared by the method and general steps described in Example 52. 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.

[0405] Example 80: 2-(4-(4-(Aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-((6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)oxy)benzonitrile (80)

[0406] Step 1: Synthesis of 5-benzyloxy-2-fluoropyridine (Compound 80-2)

[0407] Compound 80-1 (500 mg, 4.42 mmol) and K2CO3 (1.22 g, 8.84 mmol) were dissolved in dry DMF (10 mL), followed by the addition of benzyl bromide (1.13 g, 6.63 mmol). The reaction system was allowed to react at 20°C for 12 hours. After completion, the reaction was diluted with water and extracted with ethyl acetate. The organic layer was dried and concentrated, and the crude product was purified by silica gel column chromatography (mobile phase PE:EA = 92:8) to obtain compound 80-2 (885 mg). MS (ESI, m / z): 204.1 [M+H] + .

[0408] Step 2: Synthesis of 5-benzyloxy-2-(4-fluoro-1H-pyrazol-1-yl)pyridine (Compound 80-4)

[0409] 80-2 (850 mg, 4.18 mmol), 80-3 (540.01 mg, 6.27 mmol), Cs2CO3 (2.73 g 8.37 mmol), and DMF (10 mL) were added to a reaction flask and reacted at 120°C for 8 hours under nitrogen. After completion, the reaction was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic layer was dried and concentrated, and the crude product was purified by silica gel column chromatography (mobile phase PE:EA = 90:10) to obtain compound 80-4 (910 mg). MS (ESI, m / z): 270.1 [M+H] + .

[0410] Step 3: Synthesis of 6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-ol (Compound 80-5)

[0411] Compound 80-4 (890 mg, 3.31 mmol) was dissolved in MeOH (15 mL), and Pd / C (401.42 mg, 3.31 mmol) was added. The mixture was purged with hydrogen under a hydrogen balloon and allowed to react at 25°C for 2 hours. After completion of the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure to obtain compound 80-5 (560 mg). MS (ESI, m / z): 180.1 [M+H] + .

[0412] Step 4: Synthesis of tert-butyl ((7-(5-(3-chloro-6-cyano-2,5-difluorophenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (Compound 80-6)

[0413] To a reaction flask, Int-B (30 mg, 79.04 μmol), Int-A (31.72 mg, 79.04 μmol), Pd(dppf)Cl2.DCM (6.41 mg, 7.90 μmol), NaHCO3 (19.92 mg, 237.13 μmol), dioxane (5 mL), and H2O (0.5 mL) were added. The mixture was reacted at 80°C for 5 hours under nitrogen. After completion, the reaction was cooled to room temperature and concentrated under reduced pressure. The crude product was separated and purified by Prep-TLC (mobile phase DCM:MeOH = 20:1) to obtain compound 80-6 (10 mg). MS (ESI, m / z): 527.1 [M+H] + .

[0414] Step 5: Synthesis of tert-butyl ((7-(5-(3-chloro-6-cyano-2-fluoro-5-((6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)oxy)phenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (Compound 80-7)

[0415] Compound 80-5 (5.10 mg, 28.47 μmol) was dissolved in anhydrous THF (5 mL). NaH (1.14 mg, 28.47 μmol) was slowly added and stirred for 30 min. Compound 80-6 (10 mg, 18.98 μmol) was added. After N2 protection, the temperature was raised to 60°C and the reaction was allowed to proceed for 2 hr. After completion of the reaction, the temperature was lowered to room temperature and an appropriate amount of methanol was added to quench the reaction. The reaction was concentrated under reduced pressure and purified by Prep-TLC (mobile phase DCM:MeOH = 20:1) to obtain compound 80-7 (6 mg). MS (ESI, m / z): 686.1 [M+H] + .

[0416] Step 6: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-3-fluoro-6-((6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)oxy)benzonitrile (Compound 80)

[0417] Compound 80-7 (6 mg, 8.75 μmol) was added to dichloromethane (3 mL), and TFA (0.5 mL) was added dropwise. The mixture was reacted at 25°C for 2 hours. After completion of the reaction, triethylamine was added to adjust the mixture to a weak base, and the mixture was concentrated under reduced pressure. The crude product was purified by prep-HPLC to obtain compound 80 (1.5 mg). MS (ESI, m / z): 586.2 [M+H] + .

[0418] 1 H NMR (400MHz, DMSO-d6) δ12.93(s,1H),8.72(d,J=4.8Hz,1H),8.48(d,J=2.4Hz,1H),8.39(s,1H),8.21(d,J=8.4Hz,1H),8 .05–7.94(m,4H),7.91(d,J=6.0Hz,1H),7.57(dd,J=8.4,1.6Hz,1H),7.22(s,1H),6.68(s,1H),4.45(s,2H),3.88(s,3H).

[0419] Example 81: 2-(4-(4-(Aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(1-(difluoromethyl)cyclopropyloxy)-3-fluorobenzonitrile (81 and 82, 83)

[0420] Step 1: Synthesis of 4-chloro-3,6-difluoro-2-iodobenzonitrile (Compound Int-Bb)

[0421] Compound Int-Ba (70.0 g, 403 mmol) was dissolved in DCE (1.80 L), followed by the addition of Pd(OAc)2 (9.06 g, 40.3 mmol), NIS (227 g, 1.01 mol), and TFA (46.0 g, 403 mmol, 30.0 mL). The reaction was heated to 75°C and stirred for 12 hours. Two parallel batches were run according to the above steps. TLC (petroleum ether / ethyl acetate = 10:1, Rf = 0.5) confirmed complete conversion of the starting material. The reaction mixture was quenched with saturated brine (2.00 L) and then extracted with dichloromethane (2.00 L x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was then separated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 100:1 to 10:1). After concentration, compound Int-Bb (184 g) was obtained.

[0422] Step 2: Synthesis of 4-chloro-3,6-difluoro-2-(1-methyl-1H-pyrazol-5-yl)benzonitrile (Compound Int-Bc)

[0423] Int-Bb (60.0 g, 188 mmol) and 1-methyl-1H-pyrazole-5-boronic acid pinacol ester (58.8 g, 283 mmol) were dissolved in 1,4-dioxane (2.80 L) and H₂O (280 mL). Pd(dbtpf)Cl₂ (12.3 g, 18.8 mmol) and DIEA (122 g, 942 mmol, 164 mL) were then added. After replacing the nitrogen atmosphere, the reaction temperature was raised to 80°C and stirred for 1 hour. LCMS confirmed complete conversion of Int-Bb. After cooling to room temperature, the solvent was removed by distillation under reduced pressure. The mixture was then extracted with saturated brine (3.00 L) and ethyl acetate (3.00 L x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 100:1 to 4:1) to afford compound Int-Bc (25.1 g). MS m / z(ESI):254.0[M+H] + .

[0424] Step 3: Synthesis of methyl 1-(5-chloro-2-cyano-4-fluoro-3-(1-methyl-1H-pyrazol-5-yl)phenoxy)cyclopropane-1-carboxylate (Compound 81-2)

[0425] Compound Int-Bc (25.1 g, 99.0 mmol) and 81-1 (23.0 g, 198 mmol) were dissolved in THF (500 mL), and NaH (8.71 g, 218 mmol) was slowly added under ice-cooling. The reaction was then slowly warmed to room temperature and stirred for 3 hours. LCMS confirmed complete conversion. The reaction was quenched with aqueous ammonium chloride (500 mL) at 0°C, then extracted with ethyl acetate (500 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was then performed by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 100:1 to 3:1). Compound 81-2 (29.5 g) was obtained by concentration. MS m / z (ESI): 350.1 [M+H] + .

[0426] Step 4: Synthesis of 4-chloro-3-fluoro-6-(1-formylcyclopropyloxy)-2-(1-methyl-1H-pyrazol-5-yl)benzonitrile (Compound 81-3)

[0427] Preparation of LDBBA: Dissolve tert-butyl alcohol (5.26 mL, 55 mmol) in THF (25 mL). Cool to 0°C, then slowly add n-BuLi (22 mL, 2.5 M in hexane, 55 mmol) dropwise. Warm to 10°C and stir for 1 hour. Then, cool to 0°C and add DIBALH (50 mL, 1.0 M in hexane, 50 mmol) dropwise. After the addition is complete, maintain the mixture at 10°C and continue stirring for 2 hours to obtain an LDBBA solution.

[0428] The LDBBA solution was slowly added to a solution of compound 81-2 (29.5 g, 84.4 mmol) in THF (300 mL) at -65°C and maintained at -65°C for 1 hour. The reaction was complete as monitored by LCMS. The reaction was quenched by the addition of saturated aqueous ammonium chloride in an ice bath and extracted with ethyl acetate (300 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The product was separated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 100:1 to 1:1) to obtain compound 81-3 (24.7 g). MS m / z (ESI): 320.2 [M+H] + .

[0429] Step 5: Synthesis of 4-chloro-6-(1-(difluoromethyl)cyclopropyloxy)-3-fluoro-2-(1-methyl-1H-pyrazol-5-yl)benzonitrile (Compound 81-4)

[0430] Under an ice bath, DAST (29.9 g, 185 mmol, 24.5 mL) was slowly added to a solution of compound 81-3 (23.7 g, 74.1 mmol) in DCM (240 mL) and maintained at 0°C for 12 hours. LCMS monitored the reaction to be complete. The reaction mixture was then quenched with H2O (200 mL) and extracted with dichloromethane (200 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to afford compound 81-4 (27.0 g). MS m / z (ESI): 342.2 [M+H] + .

[0431] Step 6: Synthesis of 4-chloro-6-(1-(difluoromethyl)cyclopropyloxy)-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)benzonitrile (Compound 81-5)

[0432] 81-4 (27.0 g, 79.0 mmol) was dissolved in HOAc (400 mL), followed by the addition of NIS (26.7 g, 119 mmol). The reaction was heated to 80°C for 12 hours. LCMS confirmed the complete reaction. The reaction solution was then concentrated by vacuum distillation and separated by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 100:1 to 3:1). After purification and concentration, compound 81-5 (29.8 g) was obtained. MS m / z (ESI): 468.0 [M+H] + .

[0433] Step 7: Synthesis of tert-butyl ((7-(5-(3-chloro-6-cyano-5-(1-(difluoromethyl)cyclopropyloxy)-2-fluorophenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (Compound 81-6)

[0434] Compound 81-5 (8.00 g, 17.1 mmol) and Int-A (10.3 g, 25.7 mmol) were dissolved in 1,4-dioxane (160 mL) and H2O (32 mL). CsF (7.80 g) and cataCXiumA Pd G3 (1.25 g, 1.71 mmol) were then added under a nitrogen atmosphere. The reaction mixture was heated to 100°C and stirred for 12 hours. LCMS monitoring indicated product formation. After concentration, H2O (50 mL) was added, followed by extraction with ethyl acetate (100 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and separated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 100:1 to 1:1). After concentration, compound 81-6 (3.87 g) was obtained. MS m / z (ESI): 615.3 [M+H] + .

[0435] Compound 81-6 was purified by prep-SFC (chiral column: DAICEL CHIRALPAK IC (250 mm*30 mm, 10 μm); mobile phase: [CO2- i PrOH (0.1% NH3H2O)]; B%: 30%, isocratic elution mode) to give compound 83-1 (125 mg, retention time = 3.611 min), MS m / z (ESI): 615.2 [M+H] + and 82-1 (117 mg, retention time = 4.087 min), MS m / z (ESI): 615.2 [M+H] + .

[0436] Step 8: Synthesis of 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl)-4-chloro-6-(1-(difluoromethyl)cyclopropyloxy)-3-fluorobenzonitrile (Compounds 81, 82, 83)

[0437] Compound 81-6 (42 mg, 68.29 μmol) was dissolved in CH2Cl2 (2 mL), and TFA (1 mL) was added. The reaction was stirred at 25°C for 2 hours. LCMS confirmed the reaction was complete. Prep-HPLC separation and lyophilization afforded the trifluoroacetate salt of compound 81 (15.89 mg). MS (ESI, m / z): 515.1 [M+H] + .

[0438] 1 H NMR (400MHz, DMSO-d6) δ12.92(s,1H),8.43–8.35(m,4H),8.14(d,J=8.4Hz,1H),7.90(d,J=1.6Hz,1H),7.86(d,J=6 .0Hz,1H),7.44(dd,J=8.4,1.6Hz,1H),6.29(t,J=54.0Hz,1H),4.49–4.35(m,2H),3.80(s,3H),1.45–1.22(m,4H).

[0439] Compound 82-1 (110 mg, 179 μmol) was dissolved in CH2Cl2 (3 mL), and TFA (0.6 mL) was added. The reaction was stirred at 25°C for 1 hour. LCMS confirmed the reaction was complete. The product was concentrated and purified by prep-HPLC to afford compound 82 (15 mg). MS m / z (ESI): 515.3 [M+H] + .

[0440] 1 H NMR (400MHz, CD3OD) δ8.27(d,J=8.4Hz,1H),8.15(s,1H),7.81(d,J=6.0Hz,1H),7.74(d,J=1.2Hz,1H ),7.63(dd,J=8.4,1.6Hz,1H),5.93-6.29(m,1H),3.95-4.07(m,2H),3.84(s,3H),1.31-1.38(m,4H).

[0441] Compound 83-1 (117 mg, 190 μmol) was dissolved in CH2Cl2 (3 mL), and TFA (0.6 mL) was added. The reaction was stirred at 25°C for 1 hour. LCMS confirmed the reaction was complete. The product was concentrated and purified by prep-HPLC to afford compound 83 (32 mg). MS m / z (ESI): 515.3 [M+H] + .

[0442] 1 H NMR (400MHz, CD3OD) δ8.27(d,J=8.4Hz,1H),8.15(s,1H),7.81(d,J=6.0Hz,1H),7.74(s,1H),7. 63(dd,J=8.4,1.6Hz,1H),5.94-6.31(m,1H),3.95-4.08(m,2H),3.84(s,3H),1.28-1.39(m,4H).

[0443] The following compounds were prepared by the method and general steps described in Example 81. 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.

[0444] Separation method

[0445] Prep-HPLC purification of certain compounds in the examples was performed using an Aglient 1260 or Waters 2489 HPLC, with a Waters SunFire Prep C column. 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.

[0446] Biological evaluation

[0447] Experimental Example 1: Inhibitory effect of compounds on PRMT5-MTA methyltransferase activity

[0448] Experimental method 1: The prepared protein solution (PRMT5 / MEP50 and MTA mixture) was pre-incubated with different concentrations of the test compound (1000nM starting, 5-fold dilution, 7 points) at 25°C for 30 minutes, and then the prepared substrate solution (Biotinylated histone H4peptide) was added and incubated at 25°C for 90 minutes. After the reaction, the prepared detection reagent mixture (Protein A-Eu, Anti-Histone H4antibody 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.

[0449] 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:

[0450] Percent inhibition rate = (negative control Ratio - compound Ratio) / (negative control Ratio - blank control Ratio) × 100%;

[0451] 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.

[0452] Table 1. Inhibitory activity of the compounds of the present invention against PRMT5-MTA

[0453] Experimental Method 2: The inhibitory effect of the compounds of the present invention on PRMT5-MTA methyltransferase activity was determined according to the instructions of the PRMT5TR-FRET kit (BPS Bioscience). The PRMT5 / MEP50 enzyme and MTA mixture was preincubated with different concentrations of the test compound (100nM, 20nM, and 4nM) at 25°C for 30 minutes. Then, the Biotinylated histone H4 peptide / S-adenosylmethionine working solution was added and the reaction was incubated at 25°C for 120 minutes. The Eu-labeled antibody working solution was added and the reaction was slowly shaken at 25°C for 30 minutes. Then, the Dye-labeled acceptor working solution was added and the reaction was slowly shaken at 25°C for 30 minutes. The fluorescence signal ratio (Ratio) was measured using a microplate reader.

[0454] The vehicle group (DMSO) was used as the negative control, and the 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:

[0455] Percent inhibition rate = (negative control Ratio - compound Ratio) / (negative control Ratio - blank control Ratio) × 100%;

[0456] When the percentage inhibition rate was between 30% and 80%, the half-maximal inhibitory concentration (IC 50 ) or range:

[0457] IC 50 =X×(1-percent inhibition rate (%)) / percent inhibition rate (%), wherein: X is the test concentration of the compound when the inhibition rate is between 30-80%.

[0458] The inhibitory effects of the compounds on PRMT5-MTA were determined according to the above method. The results are shown in Table 2.

[0459] Table 2. Inhibitory activity of the compounds of the present invention on PRMT5-MTA

[0460] The experimental results show that the compounds of the present invention have a strong inhibitory effect on PRMT5-MTA enzyme.

[0461] Experimental Example 2: Compound Inhibition Test on MTAP Deleted / Parental HCT116 Cell Proliferation

[0462] Experimental Method 1: Further evaluate the inhibitory effect of the compounds of the present invention on cancer cell proliferation by testing their effects on cancer cell growth. In this example, MTAP Deleted / Parental HCT116 cells, obtained from Pharmaron Inc., were used.

[0463] MTAP-Deleted / Parental HCT116 cells were cultured as monolayers in MCCOYS5A medium supplemented with 10% FBS and 1% P / S at 37°C and 5% CO2. Logarithmically growing cells were digested and the concentration adjusted. 150 cells were seeded per well in a 384-well plate and cultured overnight. Prediluted compounds (10-dose starting concentration of 10,000 nM, 4-fold dilution, 10 spots) were added, or 5000 nM starting concentration of 5-fold dilution, 8 spots were added. A negative control group received DMSO, while a blank control group received culture medium. After 10 days of incubation at 37°C and 5% CO2, 40 μL of CellTiter-Glo was added to each well, and relative chemiluminescence units (RLUs) were measured using a microplate reader in chemiluminescence detection mode.

[0464] The percentage inhibition rate of compounds at different concentrations was calculated according to the following formula:

[0465] 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%

[0466] 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:

[0467] 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.

[0468] The proliferation inhibitory activity of the compounds on MTAP Deleted / Parental HCT116 cells was determined according to the above method. The results are shown in Table 3.

[0469] Table 3. Proliferation inhibitory activity of the compounds of the present invention on MTAP Deleted / Parental HCT116 cells

[0470] Experimental method 2: In this example, MTAP Deleted / Parental HCT116 cells were selected and purchased from HORIZON.

[0471] MTAP-Deleted / Parental HCT116 cells were cultured as monolayers in RPMI6140 medium supplemented with 10% FBS and 1% P / S at 37°C and 5% CO2. Cells in logarithmic growth phase were digested and the concentration adjusted. 250 cells per well were seeded into 96-well plates and cultured overnight. Pre-diluted compounds were added (5000 nM starting concentration, 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 then transferred to a 96-well opaque white plate, and the relative chemiluminescence units (RLUs) were read on a microplate reader in chemiluminescence detection mode.

[0472] The percentage inhibition rate of compounds at different concentrations was calculated according to the following formula:

[0473] 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%

[0474] 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:

[0475] 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.

[0476] The proliferation inhibitory activity of the compounds on MTAP Deleted / Parental HCT116 cells was determined according to the above method. The results are shown in Table 4.

[0477] Table 4. Proliferation inhibitory activity of the compounds of the present invention on MTAP Deleted / Parental HCT116 cells

[0478] The experimental results show that the compound of the present invention has a strong inhibitory effect on MTAP Deleted HCT116 cells and has a certain selectivity for MTAP Parental HCT116 cells.

[0479] Experimental Example 3: Pharmacokinetics of the compound in BALB / c mice

[0480] The compounds of the present invention were administered to female Balb / c mice by oral gavage (PO) to investigate their pharmacokinetic characteristics.

[0481] Compound 51 of the present invention and MRTX-1719 (prepared according to Example 16-8 of patent WO2021050915A1) were administered PO at a dose of 30 mg / kg, and the administration solvent was a 0.5% MC+0.2% Tween 80 aqueous solution. Blood samples were collected before administration (0h) and 0.25, 0.5, 1, 2, 4, 6, 8, 10, and 24h after administration; 40-50μl of blood was collected through the eye socket, placed in a K2-EDTA anticoagulant tube, centrifuged at 4000rpm for 10min (4°C), plasma was separated, and stored at -80°C. Take 10μl of plasma sample and add 90μl of acetonitrile containing internal standard, shake to mix, centrifuge at 4000rpm, 4°C for 10min, and take the supernatant for LC-MS / MS analysis. WinNonlin 6.3 software was used to calculate the pharmacokinetic parameters using a non-compartmental model. The results are shown in Table 5.

[0482] Table 5: Pharmacokinetic parameters of the compounds in mice

[0483] in conclusion:

[0484] The compound 51 of the present invention has a high exposure amount and a high blood concentration in mice after PO administration.

[0485] 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 or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, wherein the compound has the structure of formula (I): in: R 1 for R 2 and R 6 are independently selected from H, halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkoxy or 3-8 membered heterocyclic oxy; R 3 、R 4 and R 5 are each independently selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution, -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XLC 6-10 Aryl, said aryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl, -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl, -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution, -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution, or R 3 With R 4 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl, or R 4 With R 5 Together with the atoms to which it is attached, it forms a 4-11 membered spiroheterocyclyl, The condition is that when R 1 for When R 3 、R 4 and R 5 At least one selected from the above -XLC 3-8 Cycloalkyl, -XL-3-8 membered heterocycloalkyl, -XLC 6-10 Aryl, -XL-5-10 membered heteroaryl, -XL-5-10 membered bridged ring group, -XL-5-10 membered spiro ring group, 3-8 membered heterocyclic group; X is independently selected at each occurrence from O, S or NR 7 ; L is independently a direct bond or -(CH2) n -; n is 1 or 2; R 7 Selected from H, C 1-4 Alkyl or C 1-4 alkyl halide; R 8 and R 9 Independently selected from H, C 1-4 Alkyl or C 1-4 Halogenated alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, wherein: R 2 and R 6 are independently selected from H, halogen, CN, C 1-4 Alkyl, C 1-4 a haloalkyl group or a 3-8 membered heterocyclic group; Preferably, R 2 CN; R 6 is F, Cl, Br or I.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, wherein the compound has the structure of formula (IA): in: R 3 、R 4 and R 5 As defined in claim 1.

4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, wherein: R 3 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution, -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XLC 6-10 Aryl, said aryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl, -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl, -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution, -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 haloalkyl substitution; Preferably, R 3 Selected from H, 3-6 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution, -XLC 3-6 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XL-3-6 membered heterocyclic radicals, said heterocyclic radicals being optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XLC 6-10 Aryl, optionally substituted by one or more halogen, C 1-4 substituted by a haloalkyl group, a substituted or unsubstituted 5-10 membered heteroaryl group, -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted with one or more substituted or unsubstituted 5-10 membered heteroaryl, -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution, -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 haloalkyl substitution; Preferably, R 3 Selected from H, 3-6 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, hydroxyl, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic group or C 1-4 Hydroxyalkyl substitution, -XLC 3-6 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, hydroxyl, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XL-3-6 membered heterocyclic radicals, said heterocyclic radicals being optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XLC 6-10 Aryl, optionally substituted by one or more halogen, C 1-4 substituted by a haloalkyl group, a substituted or unsubstituted 5-10 membered heteroaryl group, -XL-5-10 membered heteroaryl, said heteroaryl being optionally substituted with one or more substituted or unsubstituted 5-10 membered heteroaryl, -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution, -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 haloalkyl substitution; X is independently selected at each occurrence from O or NR 7 ; L is independently selected at each occurrence from a direct bond or -(CH2)-; R 7 Selected from H or C 1-4 alkyl; R 8 and R 9 Independently selected from H, C 1-4 Alkyl or C 1-4 alkyl halide; Preferably, R 3 Selected from H, azetidine, 2-azaspiro [3.3] heptane, wherein the azetidine is replaced by one or more F, CN, CF3, methyl, methoxy, ethoxy, CHF2 or N(CH3)2 substitution; -XLC 3-6 Cycloalkyl, wherein the cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and the cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl is replaced by one or more F, hydroxyl, methyl, CHF2 or CF3 substitution; -XL-3-6 membered heterocyclyl, wherein the heterocyclyl is selected from oxetane, azetidine, tetrahydrofuran, tetrahydropyran, piperidine or pyrrolidine, and the oxetane, azetidine, tetrahydrofuran, tetrahydropyran, piperidine, pyrrolidine is optionally substituted with one or more F, -C(O)CH3, methyl, ethyl, isopropyl or tert-butyl groups; -XLC 6-10 Aryl, wherein the aryl is phenyl, wherein the phenyl is optionally substituted with one or more F, CF3, replace, -XL-5-10 membered heteroaryl, said heteroaryl being pyridyl, said pyridyl being optionally substituted with one or more replace, -XL-5-10 membered bridged ring group, wherein the bridged ring is bicyclo[1.1.1]pentane, -XL-5-10 membered spirocyclyl, wherein the spirocyclyl is selected from 7-azaspiro[3.5]nonane, spiro[3.3]heptane or 2-oxaspiro[3.3]heptane, wherein the 7-azaspiro[3.5]nonane, spiro[3.3]heptane or 2-oxaspiro[3.3]heptane is optionally substituted with one or more methyl groups; X at each occurrence is independently selected from O, NH or N(CH3); L is independently selected at each occurrence from a direct bond or -(CH2)-; Preferably, R 3 Selected from 5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, wherein: R 4 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 substituted with a haloalkoxy group or a 3-6 membered heterocyclic group, -XLC 3-8 Cycloalkyl, said cycloalkyl being substituted by one or more halogen, CN, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XL-3-8 membered heterocyclic group, said heterocyclic group is optionally substituted by one or more halogen, CN, -COCH3, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Heteroalkyl substituted, -XL-5-10 membered bridged ring group, said bridged ring group is optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 Haloalkyl substitution, -XL-5-10 membered spirocyclyl, said spirocyclyl being optionally substituted by one or more halogen, C 1-4 Alkyl, C 1-4 haloalkyl substitution; Preferably, R 4 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic substitution; Preferably, R 4 Selected from H, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic substitution; Preferably, R 4 Select H or 6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, wherein: R 5 Selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic group, said heterocyclic group being substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic substitution; Preferably, R 5 Selected from H, C 1-4 Alkyl, halogen or 3-8 membered heterocyclic group, wherein the heterocyclic group is substituted by one or more halogen, CN, -NR 8 R 9 、-COCH3、C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, 3-6 membered heterocyclic substitution; Preferably, R 5 is selected from H, methyl, F, Cl, Br or I.

7. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, wherein: The compound is selected from:

8. A pharmaceutical composition comprising a preventively or therapeutically effective amount of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof and one or more pharmaceutically acceptable carriers.

9. Use of the compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, or the pharmaceutical composition according to claim 8, 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 a cancer or tumor with MTAP deficiency; more preferably, the cancer or tumor is esophageal cancer, lung cancer, pancreatic cancer, glioblastoma, bile duct cancer, bladder cancer, mesothelioma, colorectal cancer, head and neck squamous cell carcinoma, lymphoma.

10. A method for preventing or treating a disease or condition associated with PRMT5 activity, the method comprising administering to a subject in need thereof an effective amount of the compound of any one of claims 1 to 7 or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, or the pharmaceutical composition of claim 8; Preferably, the disease or condition associated with PRMT5 activity is a cancer or tumor with MTAP deficiency; more preferably, the cancer or tumor is esophageal cancer, lung cancer, pancreatic cancer, glioblastoma, bile duct cancer, bladder cancer, mesothelioma, colorectal cancer, head and neck squamous cell carcinoma, lymphoma.

11. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotope-labeled compound thereof, or the pharmaceutical composition according to claim 8, for use in preventing or treating a disease or condition associated with PRMT5 activity; Preferably, the disease or condition associated with PRMT5 activity is a cancer or tumor with MTAP deficiency; more preferably, The cancer or tumor is preferably esophageal cancer, lung cancer, pancreatic cancer, glioblastoma, bile duct cancer, bladder cancer, mesothelioma, colorectal cancer, head and neck squamous cell carcinoma, lymphoma.

12. A method for preparing the compound according to claim 3, comprising the following steps: in: PG is an amino protecting group, preferably tert-butyloxycarbonyl; The remaining groups are as defined in claim 3; Step 1: Compounds IA-1 and IA-2 undergo substitution reaction to generate compound IA-3; Step 2: Compound IA-3 and IA-4 undergo Suzuki coupling reaction to generate compound IA-5; Step 3: Compound IA-5 is deprotected under acidic conditions to generate a compound of formula IA.

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