WO2024022186A1 - 甲基吡唑化合物、包含其的药物组合物及其制备方法和用途 - Google Patents

甲基吡唑化合物、包含其的药物组合物及其制备方法和用途 Download PDF

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WO2024022186A1
WO2024022186A1 PCT/CN2023/108081 CN2023108081W WO2024022186A1 WO 2024022186 A1 WO2024022186 A1 WO 2024022186A1 CN 2023108081 W CN2023108081 W CN 2023108081W WO 2024022186 A1 WO2024022186 A1 WO 2024022186A1
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haloalkyl
alkyl
membered
compound
group
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French (fr)
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陈忠辉
梅红江
韩晓军
田强
宋宏梅
葛均友
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四川科伦博泰生物医药股份有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4985—Pyrazines or piperazines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A61K31/502—Pyridazines; Hydrogenated pyridazines ortho- or peri-condensed with carbocyclic ring systems, e.g. cinnoline, phthalazine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A61K31/5025—Pyridazines; Hydrogenated pyridazines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D237/00—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings
    • C07D237/26—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings condensed with carbocyclic rings or ring systems
    • C07D237/30—Phthalazines
    • C07D237/32—Phthalazines with oxygen atoms directly attached to carbon atoms of the nitrogen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/10—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the present invention relates to methylpyrazole compounds, pharmaceutical compositions containing them, methods for their preparation and their use in preventing or treating diseases or conditions associated with PRMT5 activity.
  • PRMT5 Protein Arginine Methyltransferase 5
  • PRMT5 is an epigenetic enzyme and a member of the PRMT family (human bodies have PRMT1-11). It can catalyze the methylation modification of arginine residues in histones and some non-histone substrates.
  • PRMT5 is widely present in the nucleus and cytoplasm of human cells, including tissues such as heart, muscle and testis. According to the different ways of catalyzing arginine methylation, it is divided into types I, II, and III.
  • PRMT5 belongs to type II symmetric dimethylated (sDMA) PRMT, and its methyl donor is S-adenosylmethionine (SAM).
  • SAM S-adenosylmethionine
  • PRMT5 regulates the expression of multiple target proteins by catalyzing the arginine methylation of substrates, participates in multiple physiological functions, and plays an important role in tumor cell proliferation, metastasis, and malignant transformation.
  • the methylation modification of histones by PRMT5 leads to the silencing of tumor suppressor genes such as p53, ST7, NM23 and Rb, thereby promoting the occurrence and development of tumors.
  • PRMT5's regulation of non-histone proteins is mainly reflected in its influence on transcription factors (NF- ⁇ B/P65, E2F1, HoxA ⁇ GATA4), programmed cell death protein 4 (PDCD4), cell cycle and survival-related regulatory protein E2F1, and hypoxia-inducible factor Localization and expression of 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) .
  • PRMT5 can inhibit the transcription of the miR-99 family, increase FGFR3 expression, activate the Erk1/2 and Ak pathways, and lead to tumor cell growth and metastasis (Pengyu Jing, Nan Zhao, et al. Cancer Letters, 2018, 427, 38- 48).
  • PRMT5 can methylate Eif4e and FGFR3, promoting tumor cell growth (ZHANG B, DONG S, ZHU R, et al. Oncotarget, 2015, 6(26): 22799-22811.).
  • MTAP is the gene encoding methylthioadenosine phosphorylase. It is located on chromosome 9p21 and is close to the tumor suppressor gene CDKN2A (homozygous deletion often occurs). Therefore, MTAP and CDKN2A are often co-deleted in tumors (Marjon K, Kalev P, Marks K. Annual Review of Cancer Biology, 2021, 5(1)), approximately 15% of solid tumors have MTAP loss.
  • PRMT5 has two cofactors, namely activating cofactor (SAM) and inhibitory cofactor (MTA; 5'-methylthioadenosine).
  • SAM activating cofactor
  • MTA inhibitory cofactor
  • MTAP is responsible for converting MTA into Met (methionine)
  • PRMT5 is responsible for converting SAM into SAH (S-adenosyl-L-homocysteine).
  • MTA is an endogenous competitive inhibitor of PRMT5-SAM.
  • the loss of MTAP leads to the accumulation of MTA, partially inhibiting PRMT5 activity, making tumor cells more dependent on PRMT5. Under the above circumstances, inhibiting PRMT5 can further block the methylation function of PRMT5, causing tumor cell death.
  • the present invention provides new methylpyrazole compounds, which have good inhibitory effects on MTAP-deficient tumor cells and have good pharmacokinetics and other properties.
  • One aspect of the invention provides a compound of formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof:
  • R 2 and R 6 are each independently selected from H, halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 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 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1- 4- hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic group, the heterocyclic group is surrounded by one or more halogen, CN, hydroxyl, -NR 8 R 9 , -COCH 3 , C 1- 4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl or C 1-4 hydroxyalkyl substituted,
  • cycloalkyl is replaced by one or more halogen, CN, hydroxyl, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1- 4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • -XLC 6-10 aryl optionally substituted by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 Haloalkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • the heteroaryl is optionally substituted by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • R 3 and R 4 together with the atom to which they are connected form a 4-11 membered spiroheterocyclyl group
  • R 4 and R 5 together with the atom to which they are connected form a 4-11 membered spiroheterocyclyl group
  • R 1 is When, at least one of R 3 , R 4 and R 5 is selected from the above -XLC 3-8 cycloalkyl, -XL-3-8-membered heterocycloalkyl, -XLC 6-10 aryl, -XL-5- 10-membered heteroaryl group, -XL-5-10-membered bridged ring group, -XL-5-10-membered spirocyclic group, 3-8-membered heterocyclic group;
  • X is independently selected on each occurrence from O, S or NR 7 ;
  • Each occurrence of L is independently a direct bond or -(CH 2 ) n -;
  • n 1 or 2;
  • R 7 is selected from H, C 1-4 alkyl or C 1-4 haloalkyl
  • R 8 and R 9 are independently selected from H, C 1-4 alkyl or C 1-4 haloalkyl.
  • Another aspect of the invention provides a compound of formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof:
  • R 2 and R 6 are each independently selected from H, halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 Hydroxyalkyl, C 3-8 cycloalkyl, C 3-8 cycloalkoxy;
  • R 3 , R 4 and R 5 are each independently selected from H, halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1- 4 -hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, the heterocyclyl is surrounded by one or more halogens, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl base, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl substituted,
  • cycloalkyl is replaced by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkyl Oxygen, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • R 3 and R 4 together with the atom to which they are connected form a 4-11 membered spiroheterocyclyl group
  • R 4 and R 5 together with the atom to which they are connected form a 4-11 membered spiroheterocyclyl group
  • R 1 is When, at least one of R 3 , R 4 and R 5 is 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;
  • X is independently selected on each occurrence from O, S or NR 7 ;
  • Each occurrence of L is independently a direct bond or -(CH 2 ) n -;
  • n 1 or 2;
  • R 7 is selected from H, C 1-4 alkyl or C 1-4 haloalkyl
  • R 8 and R 9 are independently selected from H, C 1-4 alkyl or C 1-4 haloalkyl.
  • Another aspect of the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof and a or a variety of pharmaceutically acceptable carriers.
  • Another aspect of the invention provides a compound of the invention or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof, or a pharmaceutical composition of the invention prepared for prevention or treatment Use in medicines for diseases or conditions associated with PRMT5 activity.
  • Another aspect of the invention provides a compound of the invention or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof, or a pharmaceutical composition of the invention for prevention or treatment Diseases or conditions associated with PRMT5 activity.
  • Another aspect of the invention provides a method of preventing or treating a disease or condition associated with PRMT5 activity, the method comprising administering to an individual in need thereof an effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, a stereoisomeric compound conformers, tautomers or isotopically labeled compounds, or the pharmaceutical compositions of the invention.
  • Another aspect of the invention provides methods for preparing compounds of the invention.
  • alkyl is defined as a linear or branched saturated aliphatic hydrocarbon.
  • an alkyl group has 1 to 12, such as 1 to 4 carbon atoms.
  • 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 by one or more (such as 1 to 3) suitable substituents such as halogen (in which case the group is referred to as "Haloalkyl”) (eg CH 2 F, CHF 2 , CF 3 , CCl 3 , C 2 F 5 , C 2 Cl 5 , CH 2 CF 3 , CH 2 Cl or -CH 2 CH 2 CF 3 ,
  • halogen in which case the group is referred to as "Haloalky
  • 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 its combination.
  • Numerical ranges that may be given eg C 1-6 heteroalkyl refer to the number of carbons in the chain, including in this example 1 to 6 carbon atoms.
  • the -CH 2 OCH 2 CH 3 group is called a C 3 heteroalkyl group. Attachment to the rest of the molecule can be through a heteroatom or a carbon atom in the heteroalkyl chain.
  • haloalkyl refers to an alkyl group substituted by one or more (such as 1 to 3) the same or different halogen atoms
  • C 1-8 haloalkyl refers to haloalkyl groups with 1 to 8 carbon atoms, 1 to 6 carbon atoms and 1 to 4 carbon atoms respectively, such as -CF 3 , -C 2 F 5 , -CHF 2 , -CH 2 F, -CH 2 CF 3 , -CH 2 Cl or -CH 2 CH 2 CF 3 , etc.
  • hydroxyalkyl refers to a group in which a hydrogen atom in an alkyl group is substituted with one or more hydroxyl groups, such as a C 1-4 hydroxyalkyl group or a C 1-3 hydroxyalkyl group, Examples include, but are not limited to, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, -CH(OH) CH3 , etc.
  • alkoxy means a group in which an oxygen atom is inserted into an alkyl group (as defined above) at any reasonable position, preferably C 1-8 alkoxy, C 1-6 alkoxy group, C 1-4 alkoxy group or C 1-3 alkoxy group.
  • C 1-6 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy group, tert-butoxy, pentoxy, hexyloxy, -CH 2 -OCH 3 , etc.
  • the alkoxy group is optionally substituted by one or more (such as 1 to 3) the same or different substituents .
  • haloalkoxy means that the hydrogen atom of the alkoxy group is replaced by one or more (such as 1 to 3) the same or different halogen atoms.
  • ring or “fused ring” refers to a ring system formed by two or more cyclic structures sharing two adjacent atoms with each other.
  • the term “spiro” or “spiryl” refers to a ring system formed by two or more cyclic structures sharing a ring atom with each other.
  • the "spiro ring” may be an "aliphatic spiro ring” or a “spiro heterocyclic ring”.
  • the term “aliphatic spirocycle” refers to an all-carbon ring system formed by two or more rings sharing one carbon atom with each other, which may contain one or more double bonds, including but not limited to 5- 10-membered aliphatic spirocycle, e.g. wait.
  • spiroheterocycle refers to a ring containing one or more (eg, 1, 2, 3, or 4) heteroatoms formed from two or more saturated rings sharing one ring atom. (such as oxygen atoms, nitrogen atoms, sulfur atoms), including but not limited to 5-10-membered spiroheterocycles, 6-10-membered spiroheterocycles, 6-10-membered nitrogen-containing spiroheterocycles, 6-10-membered spiroheterocycles, Oxygen-containing spiroheterocycles, 6-10 membered sulfur-containing spiroheterocycles, etc., such as
  • the "nitrogen-containing spiroheterocycle", “oxygen-containing spiroheterocycle” and “sulfur-containing spiroheterocycle” optionally also contain one or more other heteroatoms independently selected from oxygen, nitrogen and sulfur.
  • bridged ring or “bridged cyclyl” refers to a ring system formed by two or more cyclic structures sharing two atoms that are not directly connected to each other.
  • the "bridged ring” may be a "fatty bridged ring” or a “bridged heterocyclic ring”.
  • aliphatic 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 The bond 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 rings” include but are not limited to 6-14-membered fatty bridged rings, 7-10-membered fatty bridged rings, such as wait.
  • bridged heterocycle refers to two saturated rings that share two ring atoms that are not directly connected and contain one or more (eg, 1, 2, 3, or 4) heteroatoms. (such as oxygen atoms, nitrogen atoms and/or sulfur atoms), 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-bridged heterocycles, 7-10-membered sulfur-bridged heterocycles, etc., such as wait.
  • the "nitrogen-containing bridged heterocycle", “oxygen-containing bridged heterocycle” and “sulfur-containing bridged heterocycle” optionally also contain one or more other heteroatoms independently selected from oxygen, nitrogen and sulfur.
  • cycloalkyl refers to a saturated or unsaturated non-aromatic monocyclic or fused polycyclic hydrocarbon ring radical, including but not limited to monocyclic alkyl (such as cyclopropyl, cyclobutyl , cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, etc.) and fused cycloalkyl.
  • the cycloalkyl is optionally replaced by one or more (such as 1 to 3) the same or substituted with different substituents.
  • C 3-8 cycloalkyl refers to a cycloalkyl group with 3 to 8 ring carbon atoms, such as C 3-6 cycloalkyl, which can be a monocyclic alkyl group, such as cyclopropyl, cyclobutyl , cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl, or fused cycloalkyl, such as C 5-8 fused cycloalkyl or C 5-6 fused cycloalkyl.
  • cycloalkoxy means -O-cycloalkyl, where cycloalkyl is as defined above.
  • Representative examples of cycloalkoxy include, but are not limited to, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.
  • the term "4-11 membered heterocyclyl” means a heterocyclyl containing 4-11 ring atoms, including but not limited to 4-10 membered heterocyclyl, 4-9 membered heterocyclyl, 4-8 membered heterocyclyl, 4-7 membered heterocyclyl, 5-6 membered heterocyclyl, 3-8 membered heterocyclyl, 3-7 membered heterocyclyl, 3-6 membered heterocyclyl, 4- 7-membered nitrogen-containing heterocyclic group, 4-7-membered oxygen-containing heterocyclic group, 4-7-membered sulfur-containing heterocyclic group, 5-6-membered nitrogen-containing heterocyclic group, 5-6-membered oxygen-containing heterocyclic group, 5- 6-membered sulfur-containing heterocyclic group, etc., each of the "nitrogen-containing heterocyclic group", "oxygen-containing heterocyclic group” and "sulfur-containing heterocyclic group” optionally also contains one or more independently selected from oxygen, Other heterocyclic group,
  • Examples of 4-11 membered heterocyclyl groups include, but are not limited to, oxiranyl, aziridinyl, azetidinyl, Oxetanyl, tetrahydrofuranyl, pyrrolidinyl, pyrrolidonyl (such as ), imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl.
  • heterocyclyloxy means -O-heterocyclyl, where heterocyclyl is as defined above.
  • heterocyclyl is as defined above.
  • 3-8 membered heterocyclic oxy refers to a heterocyclic oxy group having 3 to 8 heterocyclic atoms (eg, 3, 4, 5, 6, 7 or 8).
  • the heterocyclyl group can form a ring structure with a heterocyclyl group or a cycloalkyl group, and the connection point between the ring structure and other groups can be on any heterocyclyl group or cycloalkyl group.
  • Heterocyclyl also includes (but is not limited to) heterocyclyl and heterocyclyl, heterocyclyl and cycloalkyl, monoheterocyclyl and monoheterocyclyl, monoheterocyclyl and monocycloalkyl, such as 3- 7-membered (mono)heterocyclyl and 3-7-membered (mono)heterocyclyl, 3-7-membered (mono)heterocyclyl and (mono)cycloalkyl, 3-7-membered (mono)heterocyclyl and C 4-6 (mono)cycloalkyl, examples of which include but are not limited to pyrrolidinolocyclopropyl, cyclopentylaziridyl, pyrrolidinolocyclobutyl, pyrrolidinolopyrrolidinyl, Pyrrolidinopiperidyl, pyrrolidinolopiperazinyl, piperidin
  • aryl may contain 6-20 carbon atoms (C 6-20 aryl), such as 6-14 carbon atoms (C 6-14 aryl), which includes 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms; or 6-10 carbon atoms (C 6-10 aryl), which contains 6, 7, 8, 9 or 10 carbon atoms.
  • C 6-20 aryl such as 6-14 carbon atoms (C 6-14 aryl), which includes 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms; or 6-10 carbon atoms (C 6-10 aryl), which contains 6, 7, 8, 9 or 10 carbon atoms.
  • Aryl includes 6-membered aromatic carbocyclic rings, such as phenyl; bicyclic rings with at least one aromatic carbocyclic ring, such as naphthyl, indan; and tricyclic rings with at least one aromatic carbocyclic ring, such as fluorene.
  • aryl substituent is a bicyclic or tricyclic ring and at least one of the rings is a non-aromatic ring, it should be considered to be connected through an aromatic ring.
  • aryl groups include 6-membered aromatic carbocycles fused to 5-7 membered heterocycles containing one or more heteroatoms selected from N, O, and S, provided that the attachment site is an aromatic carbocycle. .
  • heteroaryl includes 5- to 10-membered (e.g., 5, 6, 7, 8, 9, or 10-membered) aromatic monocyclic rings containing from 1 to 4 (e.g., 1 2, 2, 3 or 4) heteroatoms selected from N, O and S, such as 1 to 3 heteroatoms, that is, 1, 2 or 3 heteroatoms, and the rest are carbon atoms; 8- to -12-membered bicyclic ring, the ring contains 1 to 4 (such as 1, 2, 3 or 4) heteroatoms selected from N, O and S, the rest are carbon atoms, and wherein at least one heteroatom appears in the aromatic ring; and an 11- to 14-membered tricyclic ring containing a number of 1 to 4 (such as 1, 2, 3 or 4) selected from N, O and S heteroatoms, the rest are carbon atoms, and at least one of them appears in the aromatic ring.
  • 1 to 4 e.g., 1 2, 2, 3 or 4
  • heteroatoms selected from N, O and S
  • the rest
  • halo or halogen group is defined to include F, Cl, Br or I.
  • substituted means that one or more (e.g., one, two, three or four) hydrogens on the designated atom are replaced by a selection from the indicated group, provided that no more than the designated atom is present in the case of normal valence and the substitution forms a stable compound. Combinations of substituents and/or variables are permissible only if such combinations form stable compounds.
  • substituent may be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as optionally substituted with one or more of the substituent lists, then one or more hydrogens on the carbon (to the extent of any hydrogen present) may be independently and/or together Optional substituent substitutions of choice. If the nitrogen of a substituent is described as optionally substituted with one or more of the substituents listed, then one or more hydrogens on the nitrogen (to the extent of any hydrogen present) may each be independently selected as optional. substitution of substituents.
  • each substituent is selected independently of the other.
  • each substituent may be the same as or different from another (other) substituent.
  • one or more means 1 or more than 1, such as 2, 3, 4, 5 or 10 under reasonable conditions.
  • the point of attachment of a substituent may be from any suitable position on the substituent.
  • the present invention also includes all pharmaceutically acceptable isotopically labeled compounds that are identical to the compounds of the present invention except that one or more atoms are substituted with the same atomic number but an atomic mass or mass number different from the atomic mass that predominates in nature. or atomic substitution of mass number.
  • isotopes suitable for inclusion in the compounds of the invention include, but are not limited to, isotopes of hydrogen (e.g., deuterium ( 2H ), tritium ( 3H )); isotopes of carbon Isotopes of chlorine (such as 11 C, 13 C and 14 C); chlorine isotopes (such as 36 Cl); fluorine isotopes (such as 18 F); iodine isotopes (such as 123 I and 125 I); nitrogen isotopes (such as 13 N and 15 N); oxygen isotopes (such as 15 O, 17 O and 18 O); phosphorus isotopes (such as 32 P); and sulfur isotopes (such as 35 S).
  • isotopes of hydrogen e.g., deuterium ( 2H ), tritium ( 3H )
  • isotopes of carbon Isotopes of chlorine such as 11 C, 13 C and 14 C
  • chlorine isotopes such as
  • Certain isotopically labeled compounds of the invention may be used in drug and/or substrate tissue distribution studies (eg, assays).
  • the radioactive isotopes 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 detect.
  • Substitution with positron emitting isotopes eg 11 C, 18 F, 15 O and 13 N
  • PET positron emission tomography
  • Isotopically labeled compounds of the invention may be prepared by methods analogous to those described in the accompanying Schemes and/or Examples and Preparations by using appropriate isotopically labeled reagents in place of the previously employed non-labeled reagents.
  • Pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent may be isotopically substituted, for example, D2O , acetone- d6 or DMSO- d6 .
  • stereoisomer means an isomer formed due to at least one asymmetric center.
  • compounds with one or more (e.g., one, two, three or four) asymmetric centers they can give rise to racemic mixtures, single enantiomers, diastereomeric mixtures and individual of diastereomers.
  • Certain individual molecules may also exist as geometric isomers (cis/trans).
  • compounds of the present invention may exist as mixtures of two or more structurally distinct forms in rapid equilibrium (often referred to as tautomers).
  • Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, and imine-enamine tautomers. wait.
  • nitroso-oximes can exist in equilibrium in solution as the following tautomeric forms:
  • Solid lines may be used in this article solid wedge or virtual wedge Draw the chemical bonds of the compounds of the invention.
  • the use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers at that carbon atom are included (eg, a specific enantiomer, a racemic mixture, etc.).
  • the use of solid or imaginary wedges to depict bonds to asymmetric carbon atoms is intended to demonstrate that the stereoisomers shown exist. When present in a racemic mixture, solid and imaginary wedges are used to define relative stereochemistry rather than absolute stereochemistry.
  • the compounds of the present invention are intended to exist as stereoisomers (which includes cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, They exist in the form of geometric isomers, rotamers, 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 (eg, racemic mixtures and pairs of diastereoisomers).
  • compositions of the present invention may exist in free form for therapeutic use, or, where appropriate, as pharmaceutically acceptable derivatives thereof.
  • pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, N-oxides, metabolites or prodrugs, which are administered to a patient in need thereof. After administration, the compound of the present invention or its metabolites or residues can be provided directly or indirectly. Therefore, when reference is made herein to "a compound of the invention", it is also intended to encompass the various derivative forms of the compound described above.
  • Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.
  • acid addition salts for example, hexafluorophosphate, meglumine salt, etc.
  • suitable salts see Stahl and Wermuth, "Handbook of Pharmaceutical Salts: Properties, Selection, and Use” (Wiley-VCH, 2002).
  • esters means esters derived from compounds of each general formula herein, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the free acid or alcohol form of the present invention). compound).
  • the compounds of the present invention may themselves be esters.
  • the compounds of the invention may exist in the form of solvates, preferably hydrates, wherein the compounds of the invention comprise as structural elements of the crystal lattice of the compounds a polar solvent, in particular such as water, methanol or ethanol.
  • a polar solvent in particular such as water, methanol or ethanol.
  • the amount of polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratios.
  • nitrogen-containing heterocycles are capable of forming N-oxides because nitrogen requires available lone pairs of electrons to oxidize to oxides.
  • nitrogen-containing heterocycles capable of forming N-oxides.
  • tertiary amines are capable of forming N-oxides.
  • Synthetic methods for preparing N-oxides of heterocyclic and tertiary amines are well known to those skilled in the art and include, but are not limited to, the use of peroxyacids such as peracetic acid and meta-chloroperoxybenzoic acid (MCPBA), hydrogen peroxide , alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate and dioxirane such as dimethyldioxirane to oxidize heterocyclic and tertiary amines.
  • peroxyacids such as peracetic acid and meta-chloroperoxybenzoic acid (MCPBA)
  • hydrogen peroxide alkyl hydroperoxides
  • alkyl hydroperoxides such as tert-butyl hydroperoxide
  • sodium perborate and dioxirane such as dimethyldioxirane
  • the invention also encompasses compounds of the invention containing protecting groups.
  • protecting groups In any process for preparing the compounds of the invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any relevant molecules, thereby forming chemically protected forms of the compounds of the invention. This can be accomplished by conventional protecting groups, such as those described in T. W. Greene & P. G. M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference.
  • the protecting groups can be removed at an appropriate subsequent stage using methods known in the art.
  • the invention provides a compound of Formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof:
  • R 2 and R 6 are each independently selected from H, halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 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 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1- 4 -hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic group, the heterocyclic group is surrounded by one or more halogen, CN, hydroxyl, -NR 8 R 9 , -COCH 3 , C 1- 4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl or C 1-4 hydroxyalkyl substituted,
  • cycloalkyl is replaced by one or more halogen, CN, hydroxyl, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1- 4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • -XLC 6-10 aryl optionally substituted by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 Haloalkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • the heteroaryl is optionally substituted by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • R 3 and R 4 together with the atom to which they are connected form a 4-11 membered spiroheterocyclyl group
  • R 4 and R 5 together with the atom to which they are connected form a 4-11 membered spiroheterocyclyl group
  • R 1 is When, at least one of R 3 , R 4 and R 5 is selected from the above -XLC 3-8 cycloalkyl, -XL-3-8-membered heterocycloalkyl, -XLC 6-10 aryl, -XL-5- 10-membered heteroaryl group, -XL-5-10-membered bridged ring group, -XL-5-10-membered spirocyclic group, 3-8-membered heterocyclic group;
  • X is independently selected on each occurrence from O, S or NR 7 ;
  • Each occurrence of L is independently a direct bond or -(CH 2 ) n -;
  • n 1 or 2;
  • R 7 is selected from H, C 1-4 alkyl or C 1-4 haloalkyl
  • R 8 and R 9 are independently selected from H, C 1-4 alkyl or C 1-4 haloalkyl.
  • the invention provides a compound of Formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof:
  • R 2 and R 6 are each independently selected from H, halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 Hydroxyalkyl, C 3-8 cycloalkyl, C 3-8 cycloalkoxy;
  • R 3 , R 4 and R 5 are each independently selected from H, halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1- 4 -hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, the heterocyclyl is surrounded by one or more halogens, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl base, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl substituted,
  • cycloalkyl is replaced by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkyl Oxygen, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • R 3 and R 4 together with the atom to which they are connected form a 4-11 membered spiroheterocyclyl group
  • R 4 and R 5 together with the atom to which they are connected form a 4-11 membered spiroheterocyclyl group
  • R 1 is When, at least one of R 3 , R 4 and R 5 is 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;
  • X is independently selected on each occurrence from O, S or NR 7 ;
  • Each occurrence of L is independently a direct bond or -(CH 2 ) n -;
  • n 1 or 2;
  • R 7 is selected from H, C 1-4 alkyl or C 1-4 haloalkyl
  • R 8 and R 9 are independently selected from H, C 1-4 alkyl or C 1-4 haloalkyl.
  • R 2 and R 6 are each independently selected from H, halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl or 3-8 yuan. Heterocyclic oxy group.
  • R 2 and R 6 are each independently selected from H, halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl.
  • R 2 is a 3-8 membered heterocyclic oxy group.
  • R 2 is
  • the invention provides compounds of Formula I in which R 2 is CN.
  • the invention provides compounds of Formula I in which R is F, Cl, Br or I.
  • compounds of the invention have the structure of Formula IA:
  • R 3 , R 4 and R 5 are as defined above for formula I.
  • R 3 is selected from H, halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, the heterocyclyl is replaced by one or more halogen, CN, hydroxyl, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl or C 1-4 hydroxyl Alkyl substitution,
  • cycloalkyl is replaced by one or more halogen, CN, hydroxyl, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1- 4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • -XLC 6-10 aryl optionally substituted by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 Haloalkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • the heteroaryl is optionally substituted by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl.
  • R 3 is selected from H, halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, the heterocyclyl is replaced by one or more halogen, CN, hydroxyl, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl or C 1-4 hydroxyl Alkyl substitution,
  • cycloalkyl is replaced by one or more halogen, CN, hydroxyl, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1- 4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • -XLC 6-10 aryl optionally substituted by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 Haloalkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • the heteroaryl is optionally substituted by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl;
  • Each occurrence of X is independently selected from O, S or NR 7 ;
  • L is independently selected at each occurrence from a direct bond or -(CH 2 ) n -;
  • R 7 is selected from H, C 1-4 alkyl or C 1-4 haloalkyl
  • R 8 and R 9 are independently selected from H, C 1-4 alkyl or C 1-4 haloalkyl.
  • R 3 is selected from H, halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, the heterocyclyl is replaced by one or more halogen, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl substituted,
  • cycloalkyl is replaced by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkyl Oxygen, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl.
  • R 3 is selected from H, 3-6 membered heterocyclyl, and the heterocyclyl is substituted by one or more halogen, CN, hydroxyl, - NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl or C 1- 4- hydroxyalkyl substitution,
  • cycloalkyl is replaced by one or more halogen, CN, hydroxyl, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1- 4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • -XLC 6-10 aryl optionally substituted by one or more halogens, C 1-4 haloalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • heteroaryl optionally substituted by one or more substituted or unsubstituted 5-10 membered heteroaryl
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl.
  • R 3 is selected from H, 3-6 membered heterocyclyl, and the heterocyclyl is substituted by one or more halogen, CN, hydroxyl, - NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl or C 1-4 hydroxyalkyl substituted,
  • cycloalkyl is replaced by one or more halogen, CN, hydroxyl, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1- 4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • -XLC 6-10 aryl optionally substituted by one or more halogens, C 1-4 haloalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • heteroaryl optionally substituted by one or more substituted or unsubstituted 5-10 membered heteroaryl
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl;
  • X is independently selected on each occurrence from O, S or NR 7 ;
  • L is independently selected at each occurrence from a direct bond or -(CH 2 ) n -;
  • R 7 is selected from H, C 1-4 alkyl or C 1-4 haloalkyl
  • R 8 and R 9 are independently selected from H, C 1-4 alkyl or C 1-4 haloalkyl.
  • R 3 is selected from a 3-6 membered heterocyclyl group, and the heterocyclyl group is substituted by one or more halogen, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl substituted,
  • cycloalkyl is replaced by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkyl Oxygen, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl.
  • R 3 is selected from H, 3-6 membered heterocyclyl, and the heterocyclyl is substituted by one or more halogen, CN, hydroxyl, - NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl or C 1- 4- hydroxyalkyl substitution,
  • cycloalkyl is replaced by one or more halogen, CN, hydroxyl, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1- 4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • -XLC 6-10 aryl optionally substituted by one or more halogens, C 1-4 haloalkyl, substituted or unsubstituted 5-10 membered heteroaryl,
  • heteroaryl optionally substituted by one or more substituted or unsubstituted 5-10 membered heteroaryl
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl;
  • X is independently selected from O or NR 7 on each occurrence
  • Each occurrence of L is independently selected from a direct bond or -(CH 2 )-;
  • R 7 is selected from H or C 1-4 alkyl
  • R 8 and R 9 are independently selected from H, C 1-4 alkyl or C 1-4 haloalkyl.
  • R 3 is selected from a 3-6 membered heterocyclyl group, and the heterocyclyl group is substituted by one or more halogen, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 yuan hetero Ring group substitution,
  • cycloalkyl is replaced by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkyl Oxygen, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl;
  • X is independently selected from O or NR 7 on each occurrence
  • Each occurrence of L is independently selected from a direct bond or -(CH 2 )-;
  • R 7 is selected from H or C 1-4 alkyl
  • R 8 and R 9 are independently selected from H or C 1-4 alkyl.
  • R 3 is selected from H, azetidine, 2-azaspiro[3.3]heptane, and the azetidine By one or more F, CN, CF 3 , methyl, methoxy, ethoxy, CHF 2 or N(CH 3 ) 2 substitution;
  • the cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, the cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl By one or more F, hydroxyl, methyl, CHF 2 or CF 3 substitution;
  • the heterocyclic group is selected from oxetanyl, azetidine, tetrahydrofuran, tetrahydropyran, piperidine or pyrrolidine, the oxetan base, azetidine, tetrahydrofuran, tetrahydropyran, piperidine, pyrrolidine optionally substituted by one or more F, -C(O)CH 3 , methyl, ethyl, isopropyl or tert-butyl base substitution;
  • aryl is phenyl
  • the phenyl is optionally replaced by one or more F, CF 3 , replace,
  • heteroaryl group is a pyridyl group
  • the pyridyl group is optionally substituted by one or more replace
  • bridged ring is bicyclo[1.1.1]pentane
  • the spirocyclic group is selected from 7-azaspiro[3.5]nonane, spiro[3.3]heptane or 2-oxaspiro[3.3]heptane, so The 7-azaspiro[3.5]nonane, spiro[3.3]heptane or 2-oxaspiro[3.3]heptane is optionally substituted by one or more methyl groups;
  • Each occurrence of X is independently selected from O, NH or N(CH 3 );
  • Each occurrence of L is independently selected from a direct bond or -( CH2 )-.
  • R 3 is selected from azetidine, and the azetidine is substituted by one or more F, CN, CF 3 , methane base or N(CH 3 ) 2 substitution;
  • cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, the cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl Substituted by one or more F, methyl or CF 3 ;
  • the heterocyclic group is selected from oxetanyl, azetidine, tetrahydrofuran, tetrahydropyran, piperidine or pyrrolidine, the oxetan base, azetidine, tetrahydrofuran, tetrahydropyran, piperidine, pyrrolidine optionally substituted by one or more F, -C(O)CH 3 , methyl, ethyl, isopropyl or tert-butyl base substitution;
  • the spirocyclic group is selected from 7-azaspiro[3.5]nonane, spiro[3.3]heptane or 2-oxaspiro[3.3]heptane, so The 7-azaspiro[3.5]nonane, spiro[3.3]heptane or -oxaspiro[3.3]heptane is optionally substituted by one or more methyl groups;
  • Each occurrence of X is independently selected from O, NH or N(CH 3 );
  • Each occurrence of L is independently selected from a direct bond or -( CH2 )-.
  • R 3 is selected from
  • R 3 is selected from
  • R 4 is selected from H, halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, the heterocyclyl is replaced by one or more halogen, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl substituted,
  • cycloalkyl is replaced by one or more halogen, CN, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkyl Oxygen, C 1-4 heteroalkyl substitution,
  • heterocyclyl is optionally substituted by one or more halogen, CN, -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 Alkoxy, C 1-4 haloalkoxy, C 1-4 heteroalkyl substitution,
  • the bridged ring group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl,
  • the spirocyclic group is optionally substituted by one or more halogens, C 1-4 alkyl, C 1-4 haloalkyl.
  • R 4 is selected from H, halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, the heterocyclyl is replaced by one or more halogen, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl substitution.
  • R 4 is selected from a 3-8 membered heterocyclyl group, and the heterocyclyl group is substituted by one or more halogen, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl substitution.
  • the invention provides compounds of Formula I or Formula IA in which R 4 is selected from H.
  • R 4 is
  • R 5 is selected from H, halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 hydroxyalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, the heterocyclyl is replaced by one or more halogen, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl substitution.
  • R5 is selected from H, C 1-4 alkyl, halogen or 3-8 membered heterocyclyl, and the heterocyclyl is replaced by one or Multiple halogens, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 yuan Heterocyclyl substitution.
  • R 5 is selected from halogen or 3-8 membered heterocyclyl, and the heterocyclyl is substituted by one or more halogen, CN, -NR 8 R 9 , -COCH 3 , C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 3-6 membered heterocyclyl substituted.
  • the invention provides compounds of Formula I or Formula IA in which R 5 is selected from H, C 1-4 alkyl or halogen.
  • R5 is selected from H, methyl, F, Cl, Br or I, preferably H, methyl or Cl.
  • R 5 is selected from F, Cl, Br or I, preferably Cl.
  • R3 and R4 together with the atoms to which they are attached form a 4-11 membered spiroheterocyclyl group.
  • R4 and R5 together with the atoms to which they are attached form a 4-11 membered spiroheterocyclyl group.
  • the present invention encompasses any combination of the above embodiments.
  • compounds of the invention include, but are not limited to:
  • compounds of the invention include, but are not limited to:
  • the invention provides methods for preparing compounds of Formula I-A, comprising the steps of:
  • PG is an amino protecting group, preferably tert-butoxycarbonyl
  • reaction conditions for each step are as follows:
  • the substitution reaction can be carried out in the presence of a base (such as K 2 CO 3 , Cs 2 CO 3 , DIPEA or Na 2 CO 3 ) or NaH;
  • a base such as K 2 CO 3 , Cs 2 CO 3 , DIPEA or Na 2 CO 3
  • the solvent that can be used is, for example, THF, DMF, DMSO or NMP; and the reaction Temperature is 0°C to 120°C.
  • Step 2 Compound I-A-3 and I-A-4 undergo Suzuki coupling reaction to generate compound I-A-5;
  • Catalysts that can be used in the Suzuki coupling reaction are, for example, Pd(OAc) 2 , Pd(PPh 3 ) 4 , Pd(dppf)Cl 2 , Pd(dppf)Cl 2 *DCM or cataCXiumAPdG3; bases that can be used are, for example, Cs 2 CO 3 , CsF, K 3 PO 4 , Na 2 CO 3 , KOAc, NaHCO 3 or K 2 CO 3 ; solvents that can be used are, for example, 1,4-dioxane/H 2 O or DMF/H 2 O ; and the reaction temperature is 40°C to 120°C.
  • compound I-A-5 removes the protecting group under acidic conditions to generate a compound of formula I-A;
  • the acid that can be used in this 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.
  • the invention provides pharmaceutical compositions comprising a prophylactically or therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof, and One or more pharmaceutically acceptable carriers.
  • the present invention provides pharmaceutical formulations, preferably solid, semi-solid, liquid or gaseous formulations.
  • the pharmaceutical composition or pharmaceutical formulation may also include one or more other therapeutic agents.
  • the pharmaceutical composition or pharmaceutical formulation is preferably administered by the oral, intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular or transdermal route.
  • the invention provides a compound of the invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof, or a pharmaceutical composition of the invention, or a pharmaceutical composition of the invention.
  • a pharmaceutical formulation in the manufacture of a medicament for the prevention or treatment of a disease or condition associated with PRMT5 activity.
  • the invention provides a compound of the invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof, or a pharmaceutical composition of the invention, or a pharmaceutical composition of the invention.
  • a pharmaceutical formulation in the manufacture of a medicament for modulating (eg reducing or inhibiting) PRMT5 activity.
  • the invention provides a compound of the invention, or a pharmaceutically acceptable salt, stereoisomer, tautomer or isotopically labeled compound thereof, or a pharmaceutical composition of the invention, or a pharmaceutical composition of the invention.
  • Pharmaceutical formulations for preventing or treating diseases or conditions associated with PRMT5 activity are provided.
  • the invention provides a method of 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 invention, or a pharmaceutically acceptable salt thereof, stereoisomers, tautomers or isotopically labeled compounds, or the pharmaceutical composition of the present invention, or the pharmaceutical preparation of the present invention.
  • the disease or condition associated with PRMT5 activity is preferably a MTAP-deficient cancer or tumor.
  • the cancer or tumor is preferably esophageal cancer, lung cancer, pancreatic cancer, glioblastoma, cholangiocarcinoma, bladder cancer, mesothelioma, colorectal cancer, head and neck squamous cell carcinoma, lymphoma Tumor etc.
  • pharmaceutically acceptable carrier refers to a diluent, adjuvant, excipient or vehicle that is administered with a therapeutic agent and is suitable for contact with humans and/or within the scope of reasonable medical judgment. Tissues from other animals without undue toxicity, irritation, allergic reactions, or other problems or complications commensurate with a reasonable benefit/risk ratio.
  • 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).
  • compositions of the invention may act systemically and/or locally. They may be administered by suitable routes for this purpose.
  • compositions of the present invention can be administered in suitable dosage forms.
  • Dosage regimens can be adjusted to provide the best desired response. For example, a single bolus may be administered, several divided doses may be administered over time, or the dosage may be proportionally reduced or increased as the exigencies of the therapeutic situation indicate. It is noted that dosage values may vary depending on the type and severity of the condition to be alleviated, and may include single or multiple doses. It is further understood that, for any particular individual, specific dosage regimens should be adjusted over time according to the individual needs and the professional judgment of the person administering or supervising the administration of the compositions.
  • the amount of a compound of the invention administered will depend on the individual being treated, the severity of the disorder or condition, the rate of administration, disposition of the compound, and the judgment of the prescribing physician. Generally speaking, effective doses range from about 0.0001 to about 50 mg per kilogram of body weight per day. In some cases, dosage levels no higher than the lower end of the foregoing ranges may be sufficient, while in other cases, larger dosages may still be employed without causing any deleterious side effects, provided that the larger dosage is first The dose is divided into several smaller doses to be administered throughout the day.
  • the content or amount of the compound of the present invention in the pharmaceutical composition or pharmaceutical preparation may be from about 0.01 mg to about 1000 mg.
  • treating means reversing, alleviating, or eliminating the disorder or condition to which such term applies or the progression of one or more symptoms of such disorder or condition.
  • prevention includes inhibiting and delaying the onset of a disease, and includes not only prevention before the disease develops, but also prevention of its recurrence after treatment.
  • “Individual” as used herein includes humans or non-human animals.
  • Exemplary human subjects include human subjects (referred to as patients) suffering from a disease, such as those described herein, or normal subjects.
  • non-human animals include all vertebrates, such as non-mammals (such as birds, amphibians, reptiles) and mammals, such as non-human primates, domestic animals and/or domesticated animals (such as sheep, dogs , cats, cows, pigs, etc.).
  • a pharmaceutical composition or pharmaceutical formulation of the present invention may also include one or more additional therapeutic or prophylactic agents (eg, other drugs used to treat cancer or neoplastic diseases).
  • the treatment methods of the present invention may also include administration of one or more additional therapeutic or prophylactic agents (eg, other drugs used to treat cancer or neoplastic diseases).
  • the compounds of the present invention are isolated and purified by preparative TLC, silica gel column chromatography, Prep-HPLC and/or flash column chromatography (Flash column chromatography), and their structures are confirmed by 1 H NMR and/or MS. Reaction monitoring was performed using TLC or LC-MS.
  • the 1 H NMR spectroscopy method used a Bruker superconducting nuclear magnetic resonance spectrometer (model AVACE III HD 400MHz).
  • LC/MS uses Aglient 1260 Infinity/Aglient 6120 Quadrupole.
  • TLC uses silica gel GF 254 as the stationary phase.
  • Flash column chromatography uses Biotage flash column chromatography.
  • Prep-HPLC uses Agilent 1260 model and Waters 2489 model.
  • Microwave reactions were performed using the BiotageInitiator microwave reactor.
  • reaction temperature is room temperature (15-30°C).
  • the reagents used in this application were purchased from companies such as Acros Organics, Aldrich Chemical Company, or Turbine Chemical.
  • Int-Ab (12g, 44.76mmol) was dissolved in ethanol, hydrazine hydrate (4.69g, 91.75mmol) was added, protected by nitrogen, stirred at room temperature for 30min, and then heated to 70°C for 12hr. After the reaction was completed, the mixture was cooled to room temperature, filtered with suction, and the filter cake was rinsed with ethanol and dried under vacuum to obtain the intermediate Int-Ac (8.50g). MS m/z(ESI): 282.1[M+H] + .
  • Step 4 Synthesis of 2-(7-bromo-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)isoindoline-1,3-dione (Int-Ae)
  • Step 6 Synthesis of ((7-bromo-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamic acid tert-butyl ester (Int-Ag)
  • Step 7 ((4-oxo-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-di Synthesis of tert-butyl hydrophthalazin-1-yl)methyl)carbamate (Int-A)
  • Step 4 Synthesis of 4-chloro-6-cyclopropoxy-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)benzonitrile (Int-C)
  • Step 1 4-chloro-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)-6-(oxetan-3-ylmethoxy) Synthesis of benzonitrile (1b)
  • Step 2 ((7-(5-(3-chloro-6-cyano-2-fluoro-5-(oxetan-3-ylmethoxy)phenyl)-1-methyl- Synthesis of 1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamic acid tert-butyl ester (1c)
  • Step 3 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Synthesis of )-4-chloro-3-fluoro-6-(oxetan-3-ylmethoxy)benzonitrile (1)
  • Example 1 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 from commercially purchased reagents using conventional reactions.
  • Step 1 4-chloro-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)-6-((tetrahydro-2H-pyran-4-yl) Synthesis of oxy)benzonitrile (9b)
  • Step 2 ((7-(5-(3-chloro-6-cyano-2-fluoro-5-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-1- Synthesis of methyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamic acid tert-butyl ester (9c)
  • Step 3 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Synthesis of )-4-chloro-3-fluoro-6-(tetrahydro-2H-pyran-4-yl)oxy)benzonitrile (9)
  • Example 9 The following compounds were prepared by the method and general steps described in Example 9. Other required raw materials can be purchased commercially, or synthesized by experienced synthesizers in the field of organic synthesis from commercially purchased reagents using conventional reactions.
  • Step 2 ((7-(5-(3-chloro-6-cyano-5-(3,3-difluorocyclobutoxy)-2-fluorophenyl)-1-methyl-1H- Synthesis of tert-butyl pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (15c)
  • compound 15c (40 mg) was obtained.
  • Compound 15c was resolved by SFC (column: REGIS (S, S) WHELK-O1 (250mm*25mm, 10um); mobile phase: 25% [0.1% NH 3 * H 2 O in MeOH solution] + 75% liquid CO 2 ), obtained 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] + .
  • Step 3 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Synthesis of )-4-chloro-6-(3,3-difluorocyclobutoxy)-3-fluorobenzonitrile (15/16)
  • Step 1 4-chloro-6-((3,3-dimethylcyclobutyl)methoxy)-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazole-5 -Synthesis of benzonitrile (17b)
  • Step 2 ((7-(5-(3-chloro-6-cyano-2-fluoro-5-(3,3-dimethylcyclobutyl)methoxy)phenyl)-1-methyl Synthesis of tert-butyl-1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (17c)
  • Step 3 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Synthesis of )-4-chloro-6-(3,3-dimethylcyclobutyl)methoxy)-3-fluorobenzonitrile (17)
  • Example 17 The following compounds were prepared by the method and general steps described in Example 17. Other required raw materials can be purchased commercially, or synthesized by experienced synthesizers in the field of organic synthesis from commercially purchased reagents using conventional reactions.
  • 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)
  • Step 1 4-Chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazole-5 -Synthesis of benzonitrile (21b)
  • Step 2 (7-(5-(3-chloro-6-cyano-5-(3,3-difluoroazetidin-1-yl)-2-fluorophenyl)-1-methyl Synthesis of tert-butyl (1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamate (21c)
  • Step 3 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Synthesis of )-4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluorobenzonitrile (21)
  • Step 4 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Separation of )-4-chloro-6-(3,3-difluoroazetidin-1-yl)-3-fluorobenzonitrile (21) stereoisomers compound 60 and compound 61
  • Example 21 The following compounds were prepared by the method and general steps described in Example 21. Other required raw materials can be purchased commercially, or synthesized by experienced synthesizers in the field of organic synthesis from commercially purchased reagents using conventional reactions.
  • Step 6 ((1-oxo-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2-di Synthesis of tert-butyl hydroisoquinolin-4-yl)methyl)carbamate (42g)
  • Step 7 tert-butyl ((6-(5-(3-chloro-6-cyano-5-cyclopropoxy-2-fluorophenyl)-1-methyl-1H-pyrazole-4- Synthesis of (yl)-1-oxo-1,2-dihydroisoquinolinyl)-4-yl)methyl)carbamate (42h)
  • Step 8 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydroisoquinolin-6-yl)-1-methyl-1H-pyrazole-5- Synthesis of 4-chloro-6-cyclopropyloxy-3-fluorobenzonitrile (42)
  • the first step tert-butyl ((1-oxo-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1, Synthesis of 2,3,4-tetrahydroisoquinolin-4-yl)methyl)carbamate (43b)
  • Step 2 tert-butyl ((6-(5-(3-chloro-6-cyano-5-cyclopropoxy-2-fluorophenyl)-1-methyl-1H-pyrazole-4- base)-1-oxo-1,2,3,4- Synthesis of tetrahydroisoquinolin-4-yl)methyl)carbamate (43c)
  • Step 3 2-(4-(4-(Aminomethyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)-1-methyl-1H-pyra Synthesis of azol-5-yl)-4-chloro-6-cyclopropoxy-3-fluorobenzonitrile (43)
  • 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-methylcyclopropoxy)benzonitrile (50/51)
  • Step 2 ((7-(5-(3-chloro-6-cyano-2-fluoro-5-(1-methylcyclopropoxy)phenyl)-1-methyl-1H-pyrazole) Synthesis of -4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamic acid tert-butyl ester (20c)
  • Compound 20c was separated by SFC (column: DAICEL CHIRALPAK IC (250mm*30mm, 10um); mobile phase: [Neu-ETOH]; B%: 40%-40%, A4.4; 60min), and compound 20c peak 1 was obtained (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] + .
  • Step 3 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Synthesis of )-4-chloro-3-fluoro-6-(1-methylcyclopropyloxy)benzonitrile (50/51)
  • Step 1 4-chloro-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl)-6-(3-methylazetidin-1-yl) )Synthesis of benzonitrile (52b)
  • Step 2 ((7-(5-(3-chloro-6-cyano-2-fluoro-5-(3-methylazetidin-1-yl)phenyl)-1-methyl Synthesis of -1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)formic acid tert-butyl ester (52c)
  • Step 3 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Synthesis of )-4-chloro-3-fluoro-6-(3-methylazetidin-1-yl)benzonitrile (52)
  • Example 52 The following compounds were prepared by the method and general steps described in Example 52. Other required raw materials can be purchased commercially, or synthesized by experienced synthesizers in the field of organic synthesis from commercially purchased reagents using conventional reactions.
  • 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)
  • Step 4 ((7-(5-(3-chloro-6-cyano-2,5-difluorophenyl)-1-methyl-1H-pyrazol-4-yl)-4-oxo Synthesis of -3,4-dihydrophthalazin-1-yl)methyl)carbamic acid tert-butyl ester (compound 80-6)
  • Step 5 ((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)carbamic acid tert-butyl ester ( Synthesis of compound 80-7)
  • Step 6 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Synthesis of )-4-chloro-3-fluoro-6-((6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)oxy)benzonitrile (compound 80)
  • Example 81 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl )-4-Chloro-6-(1-(difluoromethyl)cyclopropoxy)-3-fluorobenzonitrile (81 and 82,83)
  • Step 5 4-chloro-6-(1-(difluoromethyl)cyclopropoxy)-3-fluoro-2-(1-methyl-1H-pyrazol-5-yl)benzonitrile (compound Synthesis of 81-4)
  • Step 6 4-chloro-6-(1-(difluoromethyl)cyclopropyloxy)-3-fluoro-2-(4-iodo-1-methyl-1H-pyrazol-5-yl) Synthesis of benzonitrile (compound 81-5)
  • Step 7 ((7-(5-(3-chloro-6-cyano-5-(1-(difluoromethyl)cyclopropyloxy)-2-fluorophenyl)-1-methyl- Synthesis of 1H-pyrazol-4-yl)-4-oxo-3,4-dihydrophthalazin-1-yl)methyl)carbamic acid tert-butyl ester (compound 81-6)
  • Step 8 2-(4-(4-(aminomethyl)-1-oxo-1,2-dihydrophthalazin-6-yl)-1-methyl-1H-pyrazol-5-yl Synthesis of )-4-chloro-6-(1-(difluoromethyl)cyclopropyloxy)-3-fluorobenzonitrile (compounds 81 and 82,83)
  • Example 81 The following compounds were prepared by the method and general steps described in Example 81.
  • Other required raw materials can be purchased commercially, or synthesized by experienced synthesizers in the field of organic synthesis from commercially purchased reagents using conventional reactions.
  • Prep-HPLC purification of certain compounds in the examples was carried out using Aglient 1260 or Waters 2489 HPLC.
  • the separation column models were Waters SunFire Prep C 18 OBD (19mm ⁇ 150mm ⁇ 5.0 ⁇ m) and Waters Xbridge Prep C 18 OBD (19mm ⁇ 5.0 ⁇ m).
  • the column temperature is 25°C
  • the detection wavelength is 214nm, 254nm or 280nm
  • mobile phase A is acetonitrile
  • mobile phase B is 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 is adjusted according to the polarity of the compound; the mobile phase flow rate is 28 mL/min.
  • Experimental method 1 Pre-incubate the prepared protein solution (PRMT5/MEP50 and MTA mixture) with different concentrations of test compounds (1000nM starting, 5-fold dilution, 7 points) at 25°C for 30 minutes, then add the prepared Substrate solution (Biotinylated histone H4peptide), incubate the reaction at 25°C for 90 minutes; after the reaction, add the prepared detection reagent mixture (Protein A-Eu, Anti-Histone H4antibody and Streptavidin-D2), incubate at 25°C for 60 minutes, use BMG A microplate reader detects the fluorescence signal ratio (Ratio).
  • Percent inhibition rate (negative control Ratio-compound Ratio)/(negative control Ratio-blank control Ratio) ⁇ 100%;
  • Y Bottom+(Top-Bottom)/(1+10 ⁇ ((LogIC 50 -X)*Hillslope)) in GraphPad Prism 8 to detect the signal value Perform a fit and calculate the IC50 value.
  • Y is the relative inhibitory activity percentage
  • Top and Bottom are the maximum and minimum values of the fitting curve respectively
  • X is the logarithmic concentration of the compound
  • Hillslope is the slope of the curve.
  • Experimental method two Determine the inhibitory effect of the compound of the present invention on the activity of PRMT5-MTA methyltransferase according to the instructions of the PRMT5TR-FRET kit (BPS Bioscience). Pre-incubate the PRMT5/MEP50 enzyme and MTA mixture with different concentrations of test compounds (100nM, 20nM, 4nM) at 25°C for 30 minutes, then add Biotinylated histone H4 peptide/S-adenosylmethionine mixed working solution, and incubate the reaction at 25°C for 120 minutes; Add the Eu-labeled antibody working solution, shake slowly at 25°C for 30 minutes, then add the Dye-labeled acceptor working solution, shake slowly at 25°C for 30 minutes, and detect the fluorescence signal ratio (Ratio) with a microplate reader.
  • test compounds 100nM, 20nM, 4nM
  • Percent inhibition rate (negative control Ratio-compound Ratio)/(negative control Ratio-blank control Ratio) ⁇ 100%;
  • IC 50 X ⁇ (1-percent inhibition rate (%))/percent inhibition rate (%), where: X is the test concentration of the compound when the inhibition rate is between 30-80%.
  • Experimental method 1 By testing the effect of the compound of the present invention on the growth of cancer cells, the inhibitory effect of the compound of the present invention on the proliferation of cancer cells is further evaluated.
  • MTAP Deleted/Parental HCT116 cells from Pharmaron Inc. were used.
  • MTAP Deleted/Parental HCT116 cells were cultured in a monolayer in vitro.
  • the culture conditions were MCCOYS5A medium with 10% FBS+1% P/S, 37°C, and 5% CO 2 .
  • Percent inhibition rate (1-(chemiluminescence signal value of the test compound-chemiluminescence signal value of the blank control)/(chemiluminescence signal value of the negative control-chemiluminescence signal value of the blank control)) ⁇ 100%
  • 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 fitting curve respectively; x is The logarithmic concentration of a compound; and Hillslope is the slope of the curve.
  • the compound's proliferation inhibitory activity on MTAP Deleted/Parental HCT116 cells was determined according to the above method, and the results are shown in Table 3.
  • MTAP Deleted/Parental HCT116 cells were cultured in a monolayer in vitro.
  • the culture conditions were RPMI6140 medium with 10% FBS+1% P/S, 37°C, and 5% CO 2 .
  • Add 50 ⁇ l CellTiter-Glo to each well for lysis at room temperature in the dark for 10 minutes. Transfer the test solution to a 96-well opaque white plate, and then incubate the enzyme. Read the relative chemiluminescence unit value in the chemiluminescence detection mode of the standard instrument.
  • Percent inhibition rate (1-(chemiluminescence signal value of the test compound-chemiluminescence signal value of the blank control)/(chemiluminescence signal value of the negative control-chemiluminescence signal value of the blank control)) ⁇ 100%
  • 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 fitting curve respectively; x is The logarithmic concentration of a compound; and Hillslope is the slope of the curve.
  • the proliferation inhibitory activity of the compound on MTAP Deleted/Parental HCT116 cells was determined according to the above method, and the results are shown in Table 4.
  • the compound of the present invention was administered to female Balb/c mice by oral administration (PO), and the pharmacokinetic characteristics were investigated.
  • Compound 51 of the present invention and MRTX-1719 were administered PO, the dosage was 30 mg/kg, and the administration vehicle was 0.5% MC+0.2% Tween 80 aqueous solution.
  • Compound 51 of the present invention has high exposure and high blood concentration in mice after PO administration.

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Abstract

本发明涉及甲基吡唑化合物、包含其的药物组合物及其制备方法和用途。具体而言，本发明涉及式(I)化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，包含其的药物组合物，其制备方法及其制备用于预防或治疗与PRMT5活性相关的疾病或病况的药物中的用途。

Description

甲基吡唑化合物、包含其的药物组合物及其制备方法和用途
本申请要求2022年7月29日提交的题为“甲基吡唑化合物、包含其的药物组合物及其制备方法和用途”的202210903971.1号中国申请的优先权，其内容整体援引加入本文。
技术领域
本发明涉及甲基吡唑化合物、包含其的药物组合物、其制备方法及其用于预防或治疗与PRMT5活性相关的疾病或病况。
背景技术
PRMT5(Protein Arginine Methyltransferase 5)属于表观遗传酶，是PRMT家族(人体具有PRMT1-11)成员之一，可催化组蛋白和某些非组蛋白底物的精氨酸残基甲基化修饰。PRMT5广泛存在于人体细胞的细胞核和细胞质中，包括心脏、肌肉和睾丸等组织。根据催化精氨酸甲基化方式的不同分为I、II、III型，PRMT5属于II型对称双甲基化(sDMA)PRMT，其甲基供体为S-腺苷蛋氨酸(SAM)。
PRMT5通过催化底物的精氨酸甲基化，调控多种靶蛋白的表达程度，参与多种生理功能，对肿瘤细胞增殖、转移、恶性转化具有重要作用。PRMT5对组蛋白的甲基化修饰导致p53、ST7、NM23及Rb等抑癌基因的沉默，进而促进肿瘤的发生发展。PRMT5对非组蛋白的调节主要体现在影响转录因子(NF-κB/P65、E2F1、HoxA\GATA4)、程序性细胞死亡蛋白4(PDCD4)、细胞周期及存活相关调节蛋白E2F1、缺氧诱导因子1(HIF-1)、周期蛋白依赖性激酶(CDKs)、PI3K/Akt等的定位及表达(Koh CM，Bezzi M，Guccione E.Curr Mol Bio Rep,2015,1(1):19-28)。肺癌细胞中，PRMT5可抑制miR-99家族转录，增加FGFR3表达，活化Erk1/2和Ak通路，导致肿瘤细胞生长和转移(Pengyu Jing,Nan Zhao,et al.Cancer Letters,2018,427,38-48)。结肠癌中PRMT5可甲基化Eif4e和FGFR3，促进肿瘤细胞生长(ZHANG B,DONG S,ZHU R,et al.Oncotarget,2015,6(26):22799-22811.)。
MTAP是甲硫腺苷磷酸化酶的编码基因，位于染色体9p21上，与抑癌基因CDKN2A(常发生纯合性缺失)位置相近，因此MTAP常与CDKN2A在肿瘤中发生共缺失现象(Marjon K,Kalev P,Marks K.Annual Review of Cancer Biology,2021,5(1))，大约有15％的实体瘤发生MTAP缺失。
PRMT5有2个共因子(cofactor)，分别为激活性共因子(SAM)和抑制性共因子(MTA；5'-甲硫腺苷)。在正常细胞中，MTAP负责将MTA转化为Met(甲硫氨酸)，PRMT5负责将SAM转化为SAH(S-腺苷-L-高半胱氨酸)。MTA为PRMT5-SAM的内源性竞争性抑制剂，在肿瘤细胞中，MTAP的缺失导致MTA蓄积，部分抑制PRMT5活性，使得肿瘤细胞更加依赖PRMT5。在上述情况下，抑制PRMT5可进一步阻断PRMT5的甲基化功能，造成肿瘤细胞死亡。
综上所述，在MTAP缺失的情况下，抑制PRMT5显示出合成致死作用。目前尚未有PRMT5靶点的抑制剂上市。因此，需要开发新的、高效低毒的PRMT5抑制剂来满足临床需求。
发明内容
本发明提供新的甲基吡唑化合物，其对MTAP缺失的肿瘤细胞具有良好的抑制作用，并且具有良好的药物代谢动力学等性质。
本发明的一个方面提供式I的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物：
其中：
R1为
R2和R6各自独立地选自H、卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、C3-8环烷氧基或3-8元杂环氧基；
R3、R4和R5各自独立地选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
-X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
或者R3与R4连同其所连接的原子一起形成4-11元螺杂环基，
或者R4与R5连同其所连接的原子一起形成4-11元螺杂环基，
条件是，当R1为时，R3、R4和R5至少有一个选自上述-X-L-C3-8环烷基、-X-L-3-8元杂环烷基、-X-L-C6-10芳基、-X-L-5-10元杂芳基、-X-L-5-10元桥环基、-X-L-5-10元螺环基、3-8元杂环基；
X在每次出现时各自独立地选自O、S或NR7；
L在每次出现时各自独立地为直接键或-(CH2)n-；
n为1或2；
R7选自H、C1-4烷基或C1-4卤代烷基；
R8和R9独立地选自H、C1-4烷基或C1-4卤代烷基。
本发明的另一个方面提供式I的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物：
其中：
R1为
R2和R6各自独立地选自H、卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、C3-8环烷氧基；
R3、R4和R5各自独立地选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代，
-X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
或者R3与R4连同其所连接的原子一起形成4-11元螺杂环基，
或者R4与R5连同其所连接的原子一起形成4-11元螺杂环基，
条件是，当R1为时，R3、R4和R5至少有一个选自上述-X-L-C3-8环烷基、-X-L-3-8元杂环烷基、-X-L-5-10元桥环基、-X-L-5-10元螺环基、3-8元杂环基；
X在每次出现时各自独立地选自O、S或NR7；
L在每次出现时各自独立地为直接键或-(CH2)n-；
n为1或2；
R7选自H、C1-4烷基或C1-4卤代烷基；
R8和R9独立地选自H、C1-4烷基或C1-4卤代烷基。
本发明的另一方面提供药物组合物，其包含预防或治疗有效量的本发明的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物以及一种或多种药学上可接受的载体。
本发明的另一方面提供本发明的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，或者本发明的药物组合物在制备用于预防或治疗与PRMT5活性相关的疾病或病况的药物中的用途。
本发明的另一方面提供本发明的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，或者本发明的药物组合物，其用于预防或治疗与PRMT5活性相关的疾病或病况。
本发明的另一方面提供预防或治疗与PRMT5活性相关的疾病或病况的方法，所述方法包括向需要其的个体给药有效量的本发明的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，或者本发明的药物组合物。
本发明的另一方面提供制备本发明的化合物的方法。
具体实施方式
定义
除非在下文中另有定义，本文中所用的所有技术术语和科学术语的含义意图与本领域技术人员通常所理解的相同。提及本文中使用的技术意图指在本领域中通常所理解的技术，包括那些对本领域技术人员显而易见的技术的变化或等效技术的替换。虽然相信以下术语对于本领域技术人员很好理解，但仍然阐述以下定义以更好地解释本发明。
术语“包括”、“包含”、“具有”、“含有”或“涉及”及其在本文中的其它变体形式为包含性的(inclusive)或开放式的，其不排除其它未列举的元素或方法步骤，尽管其它未列举的元素或方法步骤不一定存在(即，这些术语也涵盖术语“基本上由……组成”和“由……组成”)。
如本文中所使用，术语“烷基”定义为线性或支化饱和脂肪族烃。在一些实施方案中，烷基具有1至12个，例如1至4个碳原子。例如，如本文中所使用，术语“C1-4烷基”指具有1-4个碳原子的线性或支化的脂肪族烃链(例如甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基)，其任选地被一个或多个(诸如1至3个)适合的取代基如卤素取代(此时该基团被称作“卤代烷基”)(例如CH2F、CHF2、CF3、CCl3、C2F5、C2Cl5、CH2CF3、CH2Cl或-CH2CH2CF3等)。
如本文中所使用，术语“杂烷基”指任选被取代的烷基，其具有一个或多个独立地选自除碳以外的原子的骨架链原子，例如氧、氮、硫、磷或其组合。可以给出数值范围(例如C1-6杂烷基)是指链中的碳数目，在此实例中包括1-6个碳原子。例如，-CH2OCH2CH3基团被称为C3杂烷基。与分子其余部分的连接可以通过杂烷基链中的杂原子或碳原子进行。
如本文中所使用，术语“卤代烷基”是指被一个或多个(诸如1至3个)相同或不同的卤素原子取代的烷基，术语“C1-8卤代烷基”、“C1-6卤代烷基”和“C1-4卤代烷基”分别指具有1至8个碳原子、1至6个碳原子和1-4个碳原子的卤代烷基，例如-CF3、-C2F5、-CHF2、-CH2F、-CH2CF3、-CH2Cl或-CH2CH2CF3等。
如本文中所使用，术语“羟烷基”是指烷基中的氢原子被一个或多个羟基取代所形成的基团，例如C1-4羟烷基或C1-3羟烷基，其实例包括但不限于羟甲基、羟乙基、羟丙基、羟丁基、-CH(OH)CH3等。
如本文中所使用，术语“烷氧基”意指在烷基(如上文所定义)任意合理的位置插入氧原子的基团，优选为C1-8烷氧基、C1-6烷氧基、C1-4烷氧基或C1-3烷氧基。C1-6烷氧基的代表性实例包括但不限于甲氧基、乙氧基、丙氧基、异丙氧基、正丙氧基、异丙氧基、正丁氧基、异丁氧基、叔丁氧基、戊氧基、己氧基、-CH2-OCH3等，所述烷氧基任选地被一个或多个(诸如1至3个)相同或不同的取代基取代。如术语“卤代烷氧基”是指所述烷氧基的氢原子被一个或多个(诸如1至3个)相同或不同的卤素原子取代。
如本文中所使用，术语“并环”或“稠环”指由两个或两个以上环状结构彼此共用两个相邻的原子所形成的环系。
如本文中所使用，术语“螺环”或“螺环基”指由两个或两个以上环状结构彼此共用一个环原子所形成的环系。本发明中“螺环”可以是“脂肪螺环”，也可以是“螺杂环”。如本文中所使用，术语“脂肪螺环”是指两个或两个以上环彼此共用一个碳原子所形成的全碳环系，其可以含有一个或多个双键，包括但不限于5-10元脂肪螺环，例如等。如本文中所使用，术语“螺杂环”是指由两个或两个以上饱和环共用一个环原子形成的含有一个或多个(例如1个、2个、3个或4个)杂原子(例如氧原子、氮原子、硫原子)的环状结构，包括但不限于5-10元螺杂环、6-10元螺杂环、6-10元含氮螺杂环、6-10元含氧螺杂环、6-10元含硫螺杂环等，例如 所述“含氮螺杂环”、“含氧螺杂环”、“含硫螺杂环”任选地还含有一个或多个独立地选自氧、氮、硫的其他杂原子。术语“6-10元含氮螺杂环基”是指含有共计6-10个环原子并且其中至少一个环原子为氮原子的螺杂环基。
如本文中所使用，术语“桥环”或“桥环基”指由两个或两个以上环状结构彼此共用两个不直接相连的原子所形成的环系。本发明中“桥环”可以是“脂肪桥环”，也可以是“桥杂环”。如本文中所使用，术语“脂肪桥环”是指由两个或两个以上环状结构彼此共用两个不直接相连的碳原子所形成的全碳环系，其可以含有一个或多个双键，根据组成环的数目可以分为双环、三环、四环或多环脂肪桥环，优选为双环、三环或四环，更有选为双环或三环。“脂肪桥环”包括但不限于6-14元脂肪桥环，7-10元脂肪桥环，例如等。如本文中所使用，术语“桥杂环”是指两个饱和环共用两个不直接相连的环原子形成的含有一个或多个(例如1个、2个、3个或4个)杂原子(例如氧原子、氮原子和/或硫原子)的环状结构，包括但不限于7-10元桥杂环、8-10元桥杂环、7-10元含氮桥杂环、7-10元含氧桥杂环、7-10元含硫桥杂环等，例如 等。所述“含氮桥杂环”、“含氧桥杂环”、“含硫桥杂环”任选地还含有一个或多个独立地选自氧、氮和硫的其他杂原子。
如本文中所使用，术语“环烷基”指饱和或不饱和的非芳族单环或稠合多环的烃环基，包括但不限于单环烷基(诸如环丙基、环丁基、环戊基、环己基、环庚基、环辛基、环壬基等)和稠环烷基，本发明中，环烷基任选地被一个或多个(诸如1至3个)相同或不同的取代基取代。环烷基上的碳原子任选地被氧代(oxo)基团取代(即形成C＝O)。术语“C3-8环烷基”指具有3至8个成环碳原子的环烷基，例如C3-6环烷基，其可以是单环烷基，例如环丙基、环丁基、环戊基、环己基、环庚基或环辛基，也可以是稠环烷基，例如C5-8稠环烷基或C5-6稠环烷基。
如本文中所使用，术语“环烷氧基”意指-O-环烷基，其中环烷基如上文所定义。环烷氧基的代表性实例包括但不限于环丙氧基、环丁氧基、环戊氧基、环己氧基等。
如本文中所使用，术语“杂环基”或“杂环”指具有2个或2个以上(例如3、4、5、6、7、8、9、10、11、12、13或14个)碳原子，以及一个或多个(例如1个、2个、3个或4个)杂原子的脂肪族的单环或稠合多环基团，所述杂原子包括但不限于氧原子、氮原子和硫原子，所述杂环基上的碳原子和杂原子任选地被氧代基团取代(例如形成C＝O、S(＝O)或S(＝O)2)。
如本文中所使用，术语“4-11元杂环基”意指含有4-11个环原子的杂环基，包括但不限于4-10元杂环基、4-9元杂环基、4-8元杂环基、4-7元杂环基、5-6元杂环基、3-8元杂环基、3-7元杂环基、3-6元杂环基、4-7元含氮杂环基、4-7元含氧杂环基、4-7元含硫杂环基、5-6元含氮杂环基、5-6元含氧杂环基、5-6元含硫杂环基等，所述“含氮杂环基”、“含氧杂环基”和“含硫杂环基”各自任选地还含有一个或多个独立地选自氧、氮和硫的其他杂原子。4-11元杂环基的实例包括但不限于环氧乙烷基、氮丙啶基、氮杂环丁基、 氧杂环丁基、四氢呋喃基、吡咯烷基、吡咯烷酮基(如)、咪唑烷基、吡唑烷基、四氢吡喃基、哌啶基、吗啉基、二噻烷基(dithianyl)、硫吗啉基、哌嗪基、三噻烷基(trithianyl)。
术语“杂环氧基”意指-O-杂环基，其中杂环基如上文所定义。如本文所用，“3-8元杂环氧基”指杂环原子数为3-8个(例如3、4、5、6、7或8个)的杂环氧基。
本发明中，杂环基可以与杂环基或环烷基形成并环结构，所述并环结构与其他基团的连接点可以在任一杂环基上或环烷基上，因此，本发明的杂环基还包括(但不限于)杂环基并杂环基、杂环基并环烷基、单杂环基并单杂环基、单杂环基并单环烷基，例如3-7元(单)杂环基并3-7元(单)杂环基、3-7元(单)杂环基并(单)环烷基、3-7元(单)杂环基并C4-6(单)环烷基，其实例包括但不限于吡咯烷基并环丙基、环戊基并氮杂环丙基、吡咯烷基并环丁基、吡咯烷基并吡咯烷基、吡咯烷基并哌啶基、吡咯烷基并哌嗪基、哌啶基并吗啉基、
如本文中所使用，术语“芳基”可含有6-20个碳原子(C6-20芳基)，例如6-14个碳原子(C6-14芳基)，其包含6、7、8、9、10、11、12、13或14个碳原子；或6-10个碳原子(C6-10芳基)，其包含6、7、8、9或10个碳原子。例如，苯基，萘基，茚满基，芴基等。“芳基”包括6元芳香碳环，例如苯基；至少有一个芳香碳环的双环，例如萘基、茚满和至少有一个芳香碳环的三环，例如芴。若芳基取代基为二环或三环且其中至少有一环为非芳香环，那么应认为是通过芳环联接。例如，芳基包括6元芳香碳环，这些芳香碳环与含有一个或多个选自N，O和S的杂原子的5-7元杂环稠合，条件是联接位点是芳香碳环。
如本文中所使用，术语“杂芳基”包括5元到10元(例如5、6、7、8、9或10元)的芳香单环，该环含有数目为1到4个(例如1个、2个、3个或4个)的选自N，O和S的杂原子，例如1到3个的杂原子，即1个、2个或3个杂原子，其余均为碳原子；8元到-12元双环，该环含有数目为1到4个(例如1个、2个、3个或4个)的选自N，O和S的杂原子，其余均为碳原子，且其中至少有一个杂原子出现在芳环中；和11元到14元三环，该环含有数目为1到4个(例如1个、2个、3个或4个)的选自N，O和S的杂原子，其余均为碳原子，且其中至少有一个杂原子出现在芳环中。
如本文中所使用，术语“卤代”或“卤素”基团定义为包括F、Cl、Br或I。
术语“取代”指所指定的原子上的一个或多个(例如一个、两个、三个或四个)氢被从所指出的基团的选择代替，条件是未超过所指定的原子在当前情况下的正常原子价并且所述取代形成稳定的化合物。取代基和/或变量的组合仅仅当这种组合形成稳定的化合物时才是允许的。
如果取代基被描述为“任选地被一个或多个……取代”，则取代基可(1)未被取代或(2)被取代。如果取代基的碳被描述为任选地被取代基列表中的一个或多个取代，则碳上的一个或多个氢(至存在的任何氢的程度)可单独和/或一起被独立地选择的任选的取代基替代。如果取代基的氮被描述为任选地被取代基列表中的一个或多个取代，则氮上的一个或多个氢(至存在的任何氢的程度)可各自被独立地选择的任选的取代基替代。
如果取代基被描述为“独立地选自”一组，则各取代基独立于另一者被选择。因此，各取代基可与另一(其他)取代基相同或不同。
如本文中所使用，术语“一个或多个”意指在合理条件下的1个或超过1个，例如2个、3个、4个、5个或10个。
除非指明，否则如本文中所使用，取代基的连接点可来自取代基的任意适宜位置。
当取代基的键显示为穿过环中连接两个原子的键时，则这样的取代基可键连至该可取代的环中的任一成环原子。
本发明还包括所有药学上可接受的同位素标记的化合物，其与本发明的化合物相同，除了一个或多个原子被具有相同原子序数但原子质量或质量数不同于在自然界中占优势的原子质量或质量数的原子替代。适合包含于本发明的化合物中的同位素的实例包括(但不限于)氢的同位素(例如氘(2H)、氚(3H))；碳的同位 素(例如11C、13C及14C)；氯的同位素(例如36Cl)；氟的同位素(例如18F)；碘的同位素(例如123I及125I)；氮的同位素(例如13N及15N)；氧的同位素(例如15O、17O及18O)；磷的同位素(例如32P)；及硫的同位素(例如35S)。某些同位素标记的本发明的化合物(例如掺入放射性同位素的那些)可用于药物和/或底物组织分布研究(例如分析)中。放射性同位素氚(即3H)及碳-14(即14C)因易于掺入且容易检测而特别可用于该目的。用正电子发射同位素(例如11C、18F、15O及13N)进行取代可在正电子发射断层显像术(PET)研究中用于检验底物受体占据情况。被同位素标记的本发明的化合物可通过与描述于随附路线和/或实施例及制备中的那些类似的方法通过使用适当的被同位素标记的试剂代替之前采用的非标记的试剂来制备。本发明的药学上可接受的溶剂合物包括其中结晶溶剂可被同位素取代的那些，例如，D2O、丙酮-d6或DMSO-d6。
术语“立体异构体”表示由于至少一个不对称中心形成的异构体。在具有一个或多个(例如一个、两个、三个或四个)不对称中心的化合物中，其可产生外消旋混合物、单一对映异构体、非对映异构体混合物和单独的非对映异构体。特定个别分子也可以几何异构体(顺式/反式)存在。类似地，本发明的化合物可以两种或更多种处于快速平衡的结构不同的形式的混合物(通常称作互变异构体)存在。互变异构体的代表性实例包括酮-烯醇互变异构体、苯酚-酮互变异构体、亚硝基-肟互变异构体、亚胺-烯胺互变异构体等。例如，亚硝基-肟在溶液中可以下列互变异构形式平衡存在：
要理解，本申请的范围涵盖所有这样的以任意比例(例如60％、65％、70％、75％、80％、85％、90％、95％、96％、97％、98％、99％)的异构体或其混合物。
本文中可使用实线实楔形或虚楔形描绘本发明的化合物的化学键。使用实线以描绘键连至不对称碳原子的键欲表明，包括该碳原子处的所有可能的立体异构体(例如，特定的对映异构体、外消旋混合物等)。使用实或虚楔形以描绘键连至不对称碳原子的键欲表明，存在所示的立体异构体。当存在于外消旋混合物中时，使用实及虚楔形以定义相对立体化学，而非绝对立体化学。除非另外指明，否则本发明的化合物意欲可以立体异构体(其包括顺式及反式异构体、光学异构体(例如R及S对映异构体)、非对映异构体、几何异构体、旋转异构体、构象异构体、阻转异构体及其混合物)的形式存在。本发明的化合物可表现一种以上类型的异构现象，且由其混合物(例如外消旋混合物及非对映异构体对)组成。
还应当理解，本发明的某些化合物可以游离形式存在用于治疗，或适当时，以其药学上可接受的衍生物形式存在。在本发明中，药学上可接受的衍生物包括但不限于，药学上可接受的盐、酯、溶剂合物、N-氧化物、代谢物或前药，在将它们向需要其的患者给药后，能够直接或间接提供本发明的化合物或其代谢物或残余物。因此，当在本文中提及“本发明的化合物”时，也意在涵盖化合物的上述各种衍生物形式。
本发明的化合物的药学上可接受的盐包括其酸加成盐及碱加成盐。例如六氟磷酸盐、葡甲胺盐等。适合的盐的综述参见Stahl及Wermuth的“Handbook of Pharmaceutical Salts:Properties,Selection,and Use”(Wiley-VCH,2002)。
如本文中所使用，术语“酯”意指衍生自本申请中各个通式化合物的酯，其包括生理上可水解的酯(可在生理条件下水解以释放游离酸或醇形式的本发明的化合物)。本发明的化合物本身也可以是酯。
本发明的化合物可以溶剂合物(优选水合物)的形式存在，其中本发明的化合物包含作为所述化合物晶格的结构要素的极性溶剂，特别是例如水、甲醇或乙醇。极性溶剂特别是水的量可以化学计量比或非化学计量比存在。
本领域技术人员会理解，由于氮需要可用的孤对电子来氧化成氧化物，因此并非所有的含氮杂环都能够形成N-氧化物。本领域技术人员会识别能够形成N-氧化物的含氮杂环。本领域技术人员还会认识到叔胺能够形成N-氧化物。用于制备杂环和叔胺的N-氧化物的合成方法是本领域技术人员熟知的，包括但不限于用过氧酸如过氧乙酸和间氯过氧苯甲酸(MCPBA)、过氧化氢、烷基过氧化氢如叔丁基过氧化氢、过硼酸钠和双环氧乙烷(dioxirane)如二甲基双环氧乙烷来氧化杂环和叔胺。这些用于制备N-氧化物的方法已在 文献中得到广泛描述和综述，参见例如：T.L.Gilchrist,Comprehensive Organic Synthesis,vol.7,pp 748-750；A.R.Katritzky和A.J.Boulton,Eds.,Academic Press；以及G.W.H.Cheeseman和E.S.G.Werstiuk，Advances in Heterocyclic Chemistry,vol.22,pp 390-392,A.R.Katritzky和A.J.Boulton,Eds.,Academic Press。
本发明还涵盖含有保护基的本发明的化合物。在制备本发明的化合物的任何过程中，保护在任何有关分子上的敏感基团或反应基团可能是必需的和/或期望的，由此形成本发明的化合物的化学保护的形式。这可以通过常规的保护基实现，例如，在T.W.Greene&P.G.M.Wuts,Protective Groups in Organic Synthesis,John Wiley&Sons,1991中所述的那些保护基，这些参考文献通过援引加入本文。使用本领域已知的方法，在适当的后续阶段可以移除保护基。
术语“约”是指在所述数值的±10％范围内，优选±5％范围内，更优选±2％范围内。
化合物
在一些实施方案中，本发明提供式I的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物：
其中：
R1为
R2和R6各自独立地选自H、卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、C3-8环烷氧基或3-8元杂环氧基；
R3、R4和R5各自独立地选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
-X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
或者R3与R4连同其所连接的原子一起形成4-11元螺杂环基，
或者R4与R5连同其所连接的原子一起形成4-11元螺杂环基，
条件是，当R1为时，R3、R4和R5至少有一个选自上述-X-L-C3-8环烷基、-X-L-3-8元杂环烷基、-X-L-C6-10芳基、-X-L-5-10元杂芳基、-X-L-5-10元桥环基、-X-L-5-10元螺环基、3-8元杂环基；
X在每次出现时各自独立地选自O、S或NR7；
L在每次出现时各自独立地为直接键或-(CH2)n-；
n为1或2；
R7选自H、C1-4烷基或C1-4卤代烷基；
R8和R9独立地选自H、C1-4烷基或C1-4卤代烷基。
在一些实施方案中，本发明提供式I的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物：
其中：
R1为
R2和R6各自独立地选自H、卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、C3-8环烷氧基；
R3、R4和R5各自独立地选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代，
-X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
或者R3与R4连同其所连接的原子一起形成4-11元螺杂环基，
或者R4与R5连同其所连接的原子一起形成4-11元螺杂环基，
条件是，当R1为时，R3、R4和R5至少有一个选自上述-X-L-C3-8环烷基、-X-L-3-8元杂环烷基、-X-L-5-10元桥环基、-X-L-5-10元螺环基、3-8元杂环基；
X在每次出现时各自独立地选自O、S或NR7；
L在每次出现时各自独立地为直接键或-(CH2)n-；
n为1或2；
R7选自H、C1-4烷基或C1-4卤代烷基；
R8和R9独立地选自H、C1-4烷基或C1-4卤代烷基。
在某些实施方案中，本发明提供的式I化合物中，R2和R6各自独立地选自H、卤素、CN、C1-4烷基、C1-4卤代烷基或3-8元杂环氧基。
在某些实施方案中，本发明提供的式I化合物中，R2和R6各自独立地选自H、卤素、CN、C1-4烷基、C1-4卤代烷基。
在某些实施方案中，本发明提供的式I化合物中，R2为3-8元杂环氧基。
在某些实施方案中，本发明提供的式I化合物中，R2为
在某些实施方案中，本发明提供的式I化合物中，R2为CN。
在某些实施方案中，本发明提供的式I化合物中，R6为F、Cl、Br或I。
在一些实施方案中，本发明的化合物具有式I-A所示的结构：
其中：
R3、R4和R5如上文对于式I所定义。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R3选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
-X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R3选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
-X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代；
X在每次出现时各自独立地选自O、S或NR7；
L在每次出现时各自独立地选自直接键或-(CH2)n-；
R7选自H、C1-4烷基或C1-4卤代烷基；
R8和R9独立地选自H、C1-4烷基或C1-4卤代烷基。
在另一些实施方案中，本发明提供的式I或式I-A化合物中，R3选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代，
-X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R3选自H、3-6元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
-X-L-C3-6环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-6元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、C1-4卤代烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个取代或未取代的5-10元杂芳基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R3选自H、3-6元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、 3-6元杂环基或C1-4羟烷基取代，
-X-L-C3-6环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-6元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、C1-4卤代烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个取代或未取代的5-10元杂芳基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代；
X在每次出现时各自独立地选自O、S或NR7；
L在每次出现时各自独立地选自直接键或-(CH2)n-；
R7选自H、C1-4烷基或C1-4卤代烷基；
R8和R9独立地选自H、C1-4烷基或C1-4卤代烷基。
在另一些实施方案中，本发明提供的式I或式I-A化合物中，R3选自3-6元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代，
-X-L-C3-6环烷基，所述环烷基被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-6元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R3选自H、3-6元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
-X-L-C3-6环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-6元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、C1-4卤代烷基、取代或未取代的5-10元杂芳基取代，
-X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个取代或未取代的5-10元杂芳基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代；
X在每次出现时各自独立地选自O或NR7；
L在每次出现时各自独立地选自直接键或-(CH2)-；
R7选自H或C1-4烷基；
R8和R9独立地选自H、C1-4烷基或C1-4卤代烷基。
在另一些实施方案中，本发明提供的式I或式I-A化合物中，R3选自3-6元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂 环基取代，
-X-L-C3-6环烷基，所述环烷基被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-6元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代；
X在每次出现时各自独立地选自O或NR7；
L在每次出现时各自独立地选自直接键或-(CH2)-；
R7选自H或C1-4烷基；
R8和R9独立地选自H或C1-4烷基。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R3选自H、氮杂环丁烷、2-氮杂螺[3.3]庚烷，所述的氮杂环丁烷被一个或多个F、CN、CF3、甲基、甲氧基、乙氧基、CHF2或N(CH3)2取代；
-X-L-C3-6环烷基，所述的环烷基选自环丙基、环丁基、环戊基或环已基，所述环丙基、环丁基、环戊基或环已基被一个或多个F、羟基、甲基、CHF2或CF3取代；
-X-L-3-6元杂环基，所述的杂环基选自氧杂环丁基、氮杂环丁烷、四氢呋喃、四氢吡喃、哌啶或吡咯烷，所述氧杂环丁基、氮杂环丁烷、四氢呋喃、四氢吡喃、哌啶、吡咯烷任选地被一个或多个F、-C(O)CH3、甲基、乙基、异丙基或叔丁基取代；
-X-L-C6-10芳基，所述芳基为苯基，所述苯基任选地被一个或多个F、CF3、取代，
-X-L-5-10元杂芳基，所述杂芳基为吡啶基，所述吡啶基任选地被一个或多个取代，
-X-L-5-10元桥环基，所述桥环为双环[1.1.1]戊烷，
-X-L-5-10元螺环基，所述的螺环基选自7-氮杂螺环[3.5]壬烷、螺[3.3]庚烷或2-氧杂螺[3.3]庚烷，所述7-氮杂螺环[3.5]壬烷、螺[3.3]庚烷或2-氧杂螺[3.3]庚烷任选地被一个或多个甲基取代；
X在每次出现时各自独立地选自O、NH或N(CH3)；
L在每次出现时各自独立地选自直接键或-(CH2)-。
在另一些实施方案中，本发明提供的式I或式I-A化合物中，R3选自氮杂环丁烷，所述的氮杂环丁烷被一个或多个F、CN、CF3、甲基或N(CH3)2取代；
-X-L-C3-6环烷基，所述的环烷基选自环丙基、环丁基、环戊基或环已基，所述环丙基、环丁基、环戊基或环已基被一个或多个F、甲基或CF3取代；
-X-L-3-6元杂环基，所述的杂环基选自氧杂环丁基、氮杂环丁烷、四氢呋喃、四氢吡喃、哌啶或吡咯烷，所述氧杂环丁基、氮杂环丁烷、四氢呋喃、四氢吡喃、哌啶、吡咯烷任选地被一个或多个F、-C(O)CH3、甲基、乙基、异丙基或叔丁基取代；
-X-L-5-10元螺环基，所述的螺环基选自7-氮杂螺环[3.5]壬烷、螺[3.3]庚烷或2-氧杂螺[3.3]庚烷，所述7-氮杂螺环[3.5]壬烷、螺[3.3]庚烷或-氧杂螺[3.3]庚烷任选地被一个或多个甲基取代；
X在每次出现时各自独立地选自O、NH或N(CH3)；
L在每次出现时各自独立地选自直接键或-(CH2)-。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R3选自
在另一些实施方案中，本发明提供的式I或式I-A化合物中，R3选自
在某些实施方案中，本发明提供的式I或式I-A化合物中，R4选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代，
-X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
-X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
-X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R4选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R4选自3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R4选自H。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R4为
在某些实施方案中，本发明提供的式I或式I-A化合物中，R5选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R5选自H、C1-4烷基、卤素或3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代。
在另一些实施方案中，本发明提供的式I或式I-A化合物中，R5选自卤素或3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R5选自H、C1-4烷基或卤素。
在某些实施方案中，本发明提供的式I或式I-A化合物中，R5选自H、甲基、F、Cl、Br或I，优选为H、甲基或Cl。
在另一些实施方案中，本发明提供的式I或式I-A化合物中，R5选自F、Cl、Br或I，优选为Cl。
在某些实施方案中，R3与R4连同其所连接的原子一起形成4-11元螺杂环基。
在某些实施方案中，R4与R5连同其所连接的原子一起形成4-11元螺杂环基。
本发明涵盖以上各实施方案的任意组合。
在一些实施方案中，本发明的化合物包括，但不限于：



在另一些实施方案中，本发明的化合物包括，但不限于：


制备方法
在某些实施方案中，本发明提供制备式I-A的化合物的方法，其包括以下步骤：
路线A
其中：
PG为氨基保护基，优选为叔丁氧羰基；
其余各基团如上文所定义；
各步骤的反应条件如下：
第一步：化合物I-A-1和I-A-2经取代反应生成化合物I-A-3；
所述取代反应可在碱(例如为K2CO3、Cs2CO3、DIPEA或Na2CO3)或NaH的存在下进行；可使用的溶剂例如为THF、DMF、DMSO或NMP；且反应温度为0℃至120℃。
第二步：化合物I-A-3和I-A-4经Suzuki偶联反应生成化合物I-A-5；
所述Suzuki偶联反应可使用的催化剂例如为Pd(OAc)2、Pd(PPh3)4、Pd(dppf)Cl2、Pd(dppf)Cl2*DCM或cataCXiumAPdG3；可使用的碱例如为Cs2CO3、CsF、K3PO4、Na2CO3、KOAc、NaHCO3或K2CO3；可使用的溶剂例如为1,4-二氧六环/H2O或DMF/H2O；且反应温度为40℃至120℃。
第三步：化合物I-A-5在酸性条件下脱除保护基生成式I-A的化合物；
该反应可选用的酸例如为HCl或TFA；可使用的溶剂例如为DCM、THF、EA、1,4-二氧六环或甲醇；且反应温度为20℃至80℃。
药物组合物、制剂和治疗方法
在一些实施方案中，本发明提供药物组合物，其包含预防或治疗有效量的本发明的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物以及一种或多种药学上可接受的载体。
在一些实施方案中，本发明提供药物制剂，所述药物制剂优选为固体制剂、半固体制剂、液体制剂或气态制剂。
在一些实施方案中，所述药物组合物或药物制剂还可包含一种或多种其它治疗剂。
在一些实施方案中，所述药物组合物或药物制剂优选通过口服、静脉内、动脉内、皮下、腹膜内、肌内或经皮途径给药。
在一些实施方案中，本发明提供本发明的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，或者本发明的药物组合物、或者本发明的药物制剂在制备用于预防或治疗与PRMT5活性相关的疾病或病况的药物中的用途。
在一些实施方案中，本发明提供本发明的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，或者本发明的药物组合物、或者本发明的药物制剂在制备用于调节(例如降低或抑制)PRMT5活性的药物中的用途。
在一些实施方案中，本发明提供本发明的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，或者本发明的药物组合物、或者本发明的药物制剂，其用于预防或治疗与PRMT5活性相关的疾病或病况。
在一些实施方案中，本发明提供预防或治疗与PRMT5活性相关的疾病或病况的方法，所述方法包括向需要其的个体给药有效量的本发明的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，或者本发明的药物组合物、或者本发明的药物制剂。
在一些实施方案中，所述与PRMT5活性相关的疾病或病况优选为MTAP缺失的癌症或肿瘤。
在一些实施方案中，所述癌症或肿瘤优选为食管癌、肺癌、胰腺癌、胶质母细胞瘤、胆管癌、膀胱癌、间皮瘤、结直肠癌、头颈部鳞状细胞癌、淋巴瘤等。
本发明中“药学上可接受的载体”是指与治疗剂一同给药的稀释剂、辅剂、赋形剂或媒介物，并且其在合理的医学判断的范围内适于接触人类和/或其它动物的组织而没有过度的毒性、刺激、过敏反应或与合理的益处/风险比相应的其它问题或并发症。
在本发明的药物组合物中可使用的药学上可接受的载体包括但不限于无菌液体。适合的药学上可接受的载体的实例如在Remington’s Pharmaceutical Sciences(1990)中所述。
本发明的药物组合物可以系统地作用和/或局部地作用。为此目的，它们可以适合的途径给药。
对于这些给药途径，可以适合的剂型给药本发明的药物组合物。
如本文中所使用的术语“有效量”指被给药后会在一定程度上缓解所治疗病症的一或多种症状的化合物 的量。
可调整给药方案以提供最佳所需响应。例如，可给药单次推注，可随时间给药数个分剂量，或可如治疗情况的急需所表明而按比例减少或增加剂量。要注意，剂量值可随要减轻的病况的类型及严重性而变化，且可包括单次或多次剂量。要进一步理解，对于任何特定个体，具体的给药方案应根据个体需要及给药组合物或监督组合物的给药的人员的专业判断来随时间调整。
所给药的本发明的化合物的量会取决于所治疗的个体、病症或病况的严重性、给药的速率、化合物的处置及处方医师的判断。一般而言，有效剂量在每日每kg体重约0.0001至约50mg。在一些情况下，不高于前述范围的下限的剂量水平可以是足够的，而在其它情况下，仍可在不引起任何有害副作用的情况下采用较大剂量，条件是首先将所述较大剂量分成数个较小剂量以在一整天中给药。
本发明的化合物在药物组合物或药物制剂中的含量或用量可以是约0.01mg至约1000mg。
除非另外说明，否则如本文中所使用，术语“治疗(treating)”意指逆转、减轻或清除这样的术语所应用的病症或病况或者这样的病症或病况的一或多种症状的进展。术语“预防”包括抑制和延迟疾病的发作，并且不仅包括在发展疾病之前的预防，还包括在治疗后预防疾病的复发。
如本文所使用的“个体”包括人或非人动物。示例性人个体包括患有疾病(例如本文所述的疾病)的人个体(称为患者)或正常个体。本发明中“非人动物”包括所有脊椎动物，例如非哺乳动物(例如鸟类、两栖动物、爬行动物)和哺乳动物，例如非人灵长类、家畜和/或驯化动物(例如绵羊、犬、猫、奶牛、猪等)。
在一些实施方案中，本发明的药物组合物或药物制剂还可以包含一种或多种另外的治疗剂或预防剂(例如其它用于治疗癌症或肿瘤疾病的药物)。在一些实施方案中，本发明的治疗方法还可以包括给药一种或多种另外的治疗剂或预防剂(例如其它用于治疗癌症或肿瘤疾病的药物)。
实施例
以下结合实施例进一步描述本发明，但提供这些实施例并非意图限制本发明的范围。
本文中所用的缩写具有以下含义：

本发明的化合物通过制备TLC、硅胶柱色谱法、Prep-HPLC和/或快速柱色谱法(Flash柱色谱法)来分离纯化，其结构通过1H NMR和/或MS来确证。反应监测采用TLC或LC-MS进行。
1H NMR波谱法采用Bruker超导核磁共振波谱仪(型号AVACE III HD 400MHz)。
LC/MS采用Aglient 1260 Infinity/Aglient 6120 Quadrupole。
TLC采用硅胶GF 254作为固定相。
柱色谱法一般使用200～300目硅胶(青岛海洋)作为固定相。
快速柱色谱法使用Biotage快速柱色谱仪。
Prep-HPLC采用Agilent 1260型和Waters 2489型。
微波反应使用BiotageInitiator微波反应器进行。
在以下实施例中，如无特殊说明，反应的温度为室温(15-30℃)。
本申请中所使用的试剂购自Acros Organics、Aldrich Chemical Company或特伯化学等公司。
合成实施例：
中间体Int-A：((4-氧代-7-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯
第一步：5-溴-3-((二甲氨基)亚甲基)异苯并呋喃-1(3H)-酮(Int-Ab)的合成
将Int-Aa(46g,215.93mmol)、叔丁醇钾(2.42g,21.59mmol)和DMF-DMA(165.71g,1.39mol)加入至烧瓶中，氮气保护后110℃反应36hr。反应结束后减压除去DMF-DMA，粗品经硅胶柱层析(流动相DCM：MeOH＝97:3)纯化，得到中间体Int-Ab(45.95g)。MS m/z(ESI)：268.1[M+H]+。
第二步：6-溴-4-((二甲氨基)甲基)酞嗪-1(2H)-酮(Int-Ac)的合成
将Int-Ab(12g,44.76mmol)溶于乙醇，加入水合肼(4.69g,91.75mmol)后氮气保护，室温搅拌30min后加热至70℃反应12hr。反应结束后冷却至室温，抽滤，滤饼用乙醇淋洗，真空干燥后得到中间体Int-Ac(8.50g)。MS m/z(ESI)：282.1[M+H]+。
第三步：6-溴-4-(氯甲基)酞嗪-1(2H)-酮(Int-Ad)的合成
向无水四氢呋喃(40mL)中加入Int-Ac(4g,14.18mmol)，氮气保护后于0℃缓慢滴加氯甲酸异丁酯(3.87g,28.36mmol)，加完后自然升至室温后反应16hr。反应完毕降温至0℃，搅拌下缓慢滴加0.5M稀盐酸(100mL)，始终保持反应体系温度低于10℃，滴完后抽滤，滤饼用水淋洗，干燥后得到中间体Int-Ad(3.37g)。MS m/z(ESI)：273.2[M+H]+。
第四步：2-(7-溴-4-氧-3,4-二氢酞嗪-1-基)甲基)异二氢吲哚-1,3-二酮(Int-Ae)的合成
反应瓶中加入Int-Ad(2g,7.31mmol)、酞酰亚胺钾(1.49g,8.04mmol)和DMF(27mL)，室温反应16hr。反应完毕加入6M盐酸3mL，再加入40mL水搅拌15min，抽滤，滤饼依次用碳酸氢钠水溶液和纯净水淋洗，干燥后得中间体Int-Ae(2.76g)。MS m/z(ESI)：384.2[M+H]+。
第五步：4-(氨基甲基)-6-溴酞嗪-1(2H)-酮(Int-Af)的合成
反应瓶中加入Int-Ae(2.76g,7.18mmol)、水合肼(1.44g,28.74mmol)和乙醇(30mL)，氮气保护后80℃反应2hr。反应完毕后冷至室温，抽滤，滤饼用水淋洗，干燥后得到中间体Int-Af(1.67g)。MS m/z(ESI)：254.2[M+H]+。
第六步：((7-溴-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(Int-Ag)的合成
将Int-Af(1.67g,6.57mmol)和三乙胺(2.0g,19.72mmol)加入至二氯甲烷(15mL)中，再滴加(Boc)2O(12.87g,13.15mmol)的二氯甲烷溶液(2mL)，加完后室温反应2hr。反应完毕后抽滤，滤饼干燥后得到中间体Int-Ag(1.56g)。MS m/z(ESI)：354.1[M+H]+。
第七步：((4-氧代-7-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(Int-A)的合成
将Int-Ag(747mg,2.11mmol)、Pd(dppf)Cl2(154.32mg,210.90μmol)、联硼酸频那醇酯(803.33mg,3.16mmol)和KOAc(620.93mg,6.33mmol)加入至1,4-二氧六环(8mL)中，氮气保护后100℃反应2hr。反应完毕后减压浓缩得粗品，粗品经硅胶柱层析(流动相DCM：MeOH＝95:5)纯化，得到中间体Int-A(1.10g)。MS m/z(ESI)：402.2[M+H]+。
中间体Int-B和Int-C：4-氯-3,6-二氟-2-(4-碘-1-甲基-1H-吡唑-5-基)苄腈(Int-B)和4-氯-6-环丙氧基-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)苄腈(Int-C)
第一步：4-氯-3,6-二氟-2-碘苄腈(Int-Bb)的合成
反应瓶中加入化合物Int-Ba(5.00g,28.81mmol)、Pd(OAc)2(646.83mg,2.88mmol)、NIS(12.96g,57.62mmol)、p-TsOH(2.48g,14.41mmol)和DCE(100mL)，氮气保护后75℃反应16hr。反应完毕后降至室温，加入DCM稀释，饱和食盐水洗涤，有机相经无水硫酸钠干燥，过滤后减压浓缩，粗品经硅胶柱层析(流动相PE：EA＝95:5)纯化，得到化合物Int-Bb(4.94g)。
第二步：4-氯-3,6-二氟-2-(1-甲基-1H-吡唑-5-基)苄腈(Int-Bc)的合成
反应瓶中加入化合物Int-Bb(3.00g,10.02mmol)、Pd(dtbpf)Cl2(652.96mg,1.00mmol)、1-甲基-1H-吡唑-5-硼酸频哪醇酯(5.21g,25.05mmol)、Na2CO3(2.12g,20.04mmol)、1,4-二氧六环(20mL)和H2O(4mL)，氮气保护后75℃反应16hr。反应完毕后减压浓缩，粗品经硅胶柱层析(流动相PE：EA＝78:22)纯化，得化合物Int-Bc(733mg)。MS m/z(ESI)：254.2[M+H]+。
第三步：4-氯-3,6-二氟-2-(4-碘-1-甲基-1H-吡唑-5-基)苄腈(Int-B)的合成
反应瓶中加入化合物Int-Bc(733mg,2.89mmol)、NIS(1.30g,5.78mmol)和AcOH(20mL)，氮气保护后80℃反应3hr。反应完毕后减压浓缩除去乙酸，粗品经硅胶柱层析(流动相PE：EA＝85:15)纯化，得化合物Int-B(810mg)。MS m/z(ESI)：380.1[M+H]+。
第四步：4-氯-6-环丙氧基-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)苄腈(Int-C)的合成
在冰浴下，将NaH(25.3mg,632.4μmol)加入到环丙醇(30.6mg,527.0μmol)的THF(10.0mL)溶液中，搅拌10min后加入Int-B(200.0mg,527.0μmol)，然后50℃反应1.5hr。反应完毕后加水淬灭反应，减压浓缩除去溶剂，粗品经硅胶柱层析(流动相PE：EA＝90:10)纯化，得化合物Int-C(200mg)。MS m/z(ESI)：417.9[M+H]+。
实施例1：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-(氧杂环丁烷-3-基甲氧基)苄腈(1)
第一步：4-氯-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)-6-(氧杂环丁烷-3-基甲氧基)苄腈(1b)的合成
反应瓶中加入化合物Int-B(50mg,131.74μmol)、1a(58.00mg,658μmol)、K2CO3(54.62mg,395.22μmol)和DMF(2mL)，75℃反应18hr。反应完成后降至室温，加水后用乙酸乙酯萃取，饱和食盐水洗涤，有机相经无水硫酸钠干燥，过滤后减压浓缩，粗品经硅胶柱层析(流动相PE：EA＝68:32)纯化，得到化合物1b(26mg)。MS m/z(ESI)：448.1[M+H]+。
第二步：((7-(5-(3-氯-6-氰基-2-氟-5-(氧杂环丁烷-3-基甲氧基)苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(1c)的合成
将化合物1b(26mg,50.08μmol)、Int-A(25.64mg,63.89μmol)、Pd(dppf)Cl2(4.71mg,5.81μmol)和NaHCO3(11.71mg,139.40μmol)加入到1,4-二氧六环(5mL)和H2O(0.5mL)中，氮气保护后80℃反应4hr。反应完成后降至室温减压浓缩，粗品经硅胶柱层析(DCM:MeOH＝90:10)纯化，得到化合 物1c(25mg)。MS m/z(ESI)：595.1[M+H]+。
第三步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-(氧杂环丁烷-3-基甲氧基)苄腈(1)的合成
将化合物1c(25mg,42.02μmol)加入至二氯甲烷(8mL)中，冰浴下滴加TFA(383.24mg,3.36mmol)，加完后自然升至室温反应2hr。反应完毕后加入三乙胺调至弱碱性，减压浓缩，粗产物经Pre-HPLC纯化，得到化合物1(2.20mg)。MS m/z(ESI)：495.1[M+H]+。
1H 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).
以下化合物通过实施例1所述方法和一般步骤制备得到，所需其它原料可通过商业购买，或由有机合成领域有经验的合成人员从商业购买试剂使用常规反应合成获得。



实施例9：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-(四氢-2H-吡喃-4-基)氧基)苄腈(9)
第一步：4-氯-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)-6-((四氢-2H-吡喃-4-基)氧基)苄腈(9b)的合成
冰浴下向反应瓶中加入化合物9a(26.91mg,263.48μmol)和THF(5mL)，后缓慢加入NaH(12.65mg,316.18μmol)并搅拌10min，最后滴入Int-B(100mg,263.48μmol)的THF(1mL)溶液，加完后50℃反应1hr。反应完毕降至室温，加少量甲醇淬灭，减压浓缩，粗品经硅胶柱层析(流动相DCM:MeOH＝97:3)纯化，得到化合物9b(77.0mg)。MS m/z(ESI)：462.0[M+H]+。
第二步：((7-(5-(3-氯-6-氰基-2-氟-5-((四氢-2H-吡喃-4-基)氧基)苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(9c)的合成
将化合物9b(77.0mg,166.79μmol)、Int-A(73.62mg,183.47μmol)、Pd(dppf)Cl2(13.52mg,16.68μmol)和NaHCO3(42.03mg,500.37μmol)加入到1,4-二氧六环(5mL)和H2O(0.5mL)中，氮气保护后80℃反应5hr。反应完成后降至室温，减压浓缩，粗品经硅胶柱层析(流动相PE:EA＝0:100)纯化，得到化合物9c(20.0mg)。MS m/z(ESI)：609.1[M+H]+。
第三步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-(四氢-2H-吡喃-4-基)氧基)苄腈(9)的合成
将化合物9c(20.0mg,32.84μmol)加入至二氯甲烷(10mL)中，冰浴下滴加TFA(299.54mg,2.63mmol)，自然升至室温反应2hr。反应完毕后加入三乙胺调至弱碱性，减压浓缩，粗产物经Pre-HPLC纯化，得到化合物9(4.82mg)。MS m/z(ESI)：509.1[M+H]+。
1H 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).
以下化合物通过实施例9所述方法和一般步骤制备得到，所需其它原料可通过商业购买，或由有机合成领域有经验的合成人员从商业购买试剂使用常规反应合成获得。




实施例15：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-(3,3-二氟环丁氧基)-3-氟苄腈(15/16)
第一步：4-氯-6-(3,3-二氟环丁氧基)-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)苄腈(15b)的合成
将化合物Int-B(100mg,263μmol)、15a(42.7mg,395μmol)和K2CO3(91.0mg,658μmol)加入到DMF(1mL)中，加热至80℃反应16hr。反应结束后冷至室温，加水稀释后用乙酸乙酯萃取，有机相浓缩，粗产物经Prep-TLC(PE/EA＝2/1)纯化，得到化合物15b(50mg)。MS m/z(ESI)：467.9[M+H]+。
第二步：((7-(5-(3-氯-6-氰基-5-(3,3-二氟环丁氧基)-2-氟苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(15c)的合成
将化合物15b(50.0mg,106μmol)、Int-A(60.0mg,149μmol)、CsF(97.4mg,641μmol)和cataCXiumAPdG3(7.79mg,10.6μmol)加入到1,4-二氧六环(1mL)和H2O(0.2mL)中，氮气保护后60℃反应12hr。反应完成后降至室温，加水淬灭并用乙酸乙酯萃取，饱和食盐水洗涤后无水硫酸钠干燥，过滤后减压浓缩，粗品经Prep-TLC(展开剂EA：PE＝1:1)纯化，得化合物15c(40mg)。化合物15c经过SFC拆分(柱子:REGIS(S,S)WHELK-O1(250mm*25mm,10um)；流动相:25％[0.1％NH3*H2O的MeOH溶液]+75％液体CO2)，得到化合物15c峰1(20mg)，MS m/z(ESI)：615.3[M+H]+；和化合物15c峰2(20mg)，MS m/z(ESI)：615.3[M+H]+。
第三步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-(3,3-二氟环丁氧基)-3-氟苄腈(15/16)的合成
将化合物15c峰1(20.0mg,32.5μmol)加入到HCl的EA溶液(2mL)中，室温反应1hr。反应结束后将反应液浓缩干，得到化合物15(14.7mg)。MS m/z(ESI)：515.2[M+H]+。
1H 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).
将化合物15c峰2(20.0mg,32.5μmol)加入到HCl的EA溶液(2mL)中，室温反应1hr。反应结 束后将反应液浓缩干，得到化合物16(14.9mg)。MS m/z(ESI)：515.1[M+H]+。
1H 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).
实施例17：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-(3,3-二甲基环丁基)甲氧基)-3-氟苄腈(17)
第一步：4-氯-6-((3,3-二甲基环丁基)甲氧基)-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)苄腈(17b)的合成
冰浴下向反应瓶中加入化合物17a(21.7mg,190μmol)和THF(5mL)，后缓慢加入NaH(9.5mg,237μmol)并搅拌30min，最后滴入Int-B(60.0mg,158μmol)的THF(1mL)溶液，加完后25℃反应2hr。反应完毕降至室温，加少量水淬灭，减压浓缩，粗品经硅胶柱层析(流动相PE:EA＝5:1)纯化，得到化合物17b(50.0mg)。MS m/z(ESI)：473.9[M+H]+。
第二步：((7-(5-(3-氯-6-氰基-2-氟-5-(3,3-二甲基环丁基)甲氧基)苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(17c)的合成
将化合物17b(38.5mg,81.3μmol)、Int-A(42.4mg,106μmol)、CsF(74.1mg,488umol)和cataCXium A Pd-G3(5.9mg,8.13umol)加入到1,4-二氧六环(10mL)和H2O(2.5mL)中，氮气保护后60℃反应12hr。反应完成后降至室温，加水淬灭反应，然后乙酸乙酯萃取，合并有机相，饱和食盐水洗涤，有机相用无水硫酸钠干燥，抽滤，浓缩，粗品经prep-HPLC(流动相H2O(NH4HCO3)/ACN＝1:1)纯化，得到化合物17c(33.0mg)。MS m/z(ESI)：621.6[M+H]+。
第三步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-(3,3-二甲基环丁基)甲氧基)-3-氟苄腈(17)的合成
在冰浴下将化合物17c(30.0mg,48.3μmol)加入至HCl/EtOAc(4M,5.00mL)中，自然升至室温反应1hr。反应完毕后水淬灭反应，然后乙酸乙酯萃取三次，合并有机相，用饱和食盐水洗涤，有机相用无水硫酸钠干燥，抽滤，减压浓缩，粗产物经Pre-HPLC纯化，得到化合物17(7.00mg)。MS m/z(ESI)：521.0[M+H]+。
1H 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),4.44-4.35(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).
以下化合物通过实施例17所述方法和一般步骤制备得到，所需其它原料可通过商业购买，或由有机合成领域有经验的合成人员从商业购买试剂使用常规反应合成获得。



实施例21：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-(3,3-二氟氮杂环丁烷-1-基)-3-氟苄腈(21和60,61)
第一步：4-氯-6-(3,3-二氟氮杂环丁烷-1-基)-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)苄腈(21b)合成
将化合物Int-B(50mg,131.74μmol)、21a(51mg,395.22μmol)和DIPEA(85mg,658.71μmol)加入到NMP(1.5mL)中，微波加热100℃反应8hr。反应完成后降至室温，加水淬灭并用乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤后减压浓缩，粗品经硅胶柱层析(流动相EA：PE＝15:85)纯化，得化合物21b(35mg)。
第二步：(7-(5-(3-氯-6-氰基-5-(3,3-二氟氮杂环丁烷-1-基)-2-氟苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(21c)的合成
将化合物21b(35mg,77.33μmol)、Int-A(34mg,85.06μmol)、Pd(dppf)Cl2(6mg,7.73μmol)和NaHCO3(19mg,231.99μmol)加入到1,4-二氧六环(3mL)和H2O(0.5mL)中，氮气保护后80℃反应5hr。反应完成后降至室温，加水淬灭并用乙酸乙酯萃取，饱和食盐水洗涤后无水硫酸钠干燥，过滤后减压浓缩，粗品经Prep-TLC(展开剂EA：PE＝1:1)纯化，得化合物21c(20mg)。MS m/z(ESI)：601.3[M+H]+。
第三步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-(3,3-二氟氮杂环丁烷-1-基)-3-氟苄腈(21)的合成
将化合物21c(25mg,38.12μmol)加入至二氯甲烷(1.5mL)中，再滴加TFA(0.5mL)，氮气保护后室温反应2hr。反应完毕后减压浓缩除去溶剂，再用DCM溶解后加入三乙胺调至弱碱性，减压浓缩，粗产物经Pre-HPLC纯化，得到化合物21(19.05mg)。MS m/z(ESI)：500.1[M+H]+。
1H 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,1H),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).
第四步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-(3,3-二氟氮杂环丁烷-1-基)-3-氟苄腈(21)立体异构体化合物60和化合物61的分离
化合物21经过prep-SFC(手性柱:DAICEL CHIRALPAK IC(250mm*30mm,10um)；流动相:[CO2-EtOH(0.05％DEA)])拆分得到化合物60(8.78mg,保留时间＝1.073min)，MS m/z(ESI):500.1[M+H]+和61(13.48mg,保留时间＝1.730min)，MS m/z(ESI):500.1[M+H]+。
化合物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)。
化合物61：1H 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)。
以下化合物通过实施例21所述方法和一般步骤制备得到，所需其它原料可通过商业购买，或由有机合成领域有经验的合成人员从商业购买试剂使用常规反应合成获得。

实施例42：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢异喹啉-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-环丙氧基-3-氟苄腈(42)
第一步：4-溴-2-(溴甲基)苯甲酸甲酯(42b)合成
将化合物42a(10.0g,43.6mmol)溶于CCl4(100mL)中，然后加入BPO(528mg,2.18mmol)和NBS(9.32g,52.4mmol)，在氮气氛围下加热至80℃反应12hr。反应完成后降至室温，加水淬灭并用二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤后减压浓缩，粗品经HPLC(流动相0.1％FA)纯化，得化合物42b(9.8g)。
第二步：4-溴-2-(氰基甲基)苯甲酸乙酯(42c)的合成
在0℃将NaCN(910mg，18.5mmol)的水(10mL)溶液加入到化合物42b(5.0g,16.2mmol)的乙醇(50mL)溶液中，氮气保护后80℃反应2hr。反应完成后降至室温，加水淬灭并用乙酸乙酯萃取，饱和食盐水洗涤后无水硫酸钠干燥，过滤后减压浓缩，粗品经硅胶柱层析(展开剂PE:EA＝0:1)纯化，得化合物42c(1.2g)。
第三步：6-溴-1-氧代-1,2-二氢异喹啉-4-甲腈(42d)的合成
将化合物42c(1.2g,4.48mmol)和1,3,5-三嗪(435mg,5.37mmol)溶于MeOH(20mL)中，再加入NaOMe(1.21g,223.mmol)的MeOH(5mL)溶液，氮气保护后25℃反应20min。反应完毕后加入NH4Cl水溶液(50mL)淬灭反应，加入乙酸乙酯稀释并在25℃搅拌20min，抽滤收集滤饼，得到化合物42d(900.0mg)。
第四步：1-氧代-6-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1,2-二氢异喹啉-4-甲腈(42e)的合成
将化合物42d(200mg,80.3μmol),B2Pin2(305mg,1.20mmol),Pd(dppf)Cl2(58.7mg,80.3μmol),KOAc(157mg,1.61mmol)溶于1,4-二氧六环(3mL)中，氮气保护后80℃反应12hr。反应完毕后直接浓缩，经过硅胶柱层析(展开剂PE:EA＝1:1)纯化后，得到化合物42e(180.0mg),MS m/z(ESI)：214.5[M-81]+。
第五步：4-(氨甲基)-6-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)异喹啉-1(2H)-酮(42f)的合成
将化合物42e(50.0mg,168μmol)加入到Raney-Ni(14.4mg,168μmol)的NH3/MeOH(1mL)溶液中，在H2(15psi)氛围下20℃反应12hr。反应完毕后直接抽滤，得到化合物42f(25.0mg),MS m/z(ESI)：219.0[M-81]+。
第六步：((1-氧代-6-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1,2-二氢异喹啉-4-基)甲基)氨基甲酸叔丁酯(42g)的合成
将化合物42f(25.0mg,83μmol)溶于CH2Cl2(0.5mL)中，加入Boc2O(36.3mg,166μmol)和TEA(25.2mg,249μmol)，置换氮气后在25℃反应10min。直接浓缩后用石油醚(2mL)洗涤，得到化合物42g(15.0mg),MS m/z(ESI)：401.0[M+H]+。
第七步：叔丁基((6-(5-(3-氯-6-氰基-5-环丙氧基-2-氟苯基)-1-甲基-1H-吡唑-4-基)-1-氧代-1,2-二氢异喹啉)-4-基)甲基)氨基甲酸酯(42h)的合成
将化合物42g(15.0mg,37.4μmol)、Int-C(15.6mg,37.4μmol)、CsF(34.1mg,224μmol)和cataCXium A Pd-G3(2.7mg,3.7μmol)加入到1,4-二氧六环(0.5mL)和H2O(0.1mL)中，氮气保护后60℃反应12hr。反应完成后降至室温，加水淬灭反应，然后乙酸乙酯萃取，合并有机相，饱和食盐水洗涤，有机相用无水硫酸钠干燥，抽滤，浓缩，粗品经prep-HPLC(流动相PE/EA＝1:1)纯化，得到化合物42h(10.0mg)。MS m/z(ESI)：564.2[M+H]+。
第八步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢异喹啉-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-环丙氧基-3-氟苄腈(42)的合成
在冰浴下将化合物42h(10.0mg,48.3μmol)加入至HCl/EtOAc(4M,5.00mL)中，然后升温至25℃反应30min。反应完毕后水淬灭反应，然后乙酸乙酯萃取三次，合并有机相，用饱和食盐水洗涤，有机相用无水硫酸钠干燥，抽滤，减压浓缩，粗产物经Pre-HPLC([water(HCl)-ACN]；B％:12％-42％)纯化，得到化合物42(2.47mg)。MS m/z(ESI)：464.2[M+H]+。
1H 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)
实施例43：2-(4-(4-(氨基甲基)-1-氧代-1,2,3,4-四氢异喹啉-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-环丙氧基-3-氟苄腈(43)
第一步：叔丁基((1-氧代-6-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1,2,3,4-四氢异喹啉-4-基)甲基)氨基甲酸酯(43b)的合成
将化合物42g(10.0mg,24.9μmol)溶于EtOH(2mL)中，加入Pd/C(0.1g,10％purity)，在H2(50psi)后在50℃反应12hr。直接抽滤，得到化合物43b(15.0mg),MS m/z(ESI)：347.0[M-55]+。
第二步：叔丁基((6-(5-(3-氯-6-氰基-5-环丙氧基-2-氟苯基)-1-甲基-1H-吡唑-4-基)-1-氧代-1,2,3,4- 四氢异喹啉-4-基)甲基)氨基甲酸酯(43c)的合成
将化合物43b(15.0mg,37.3μmol)、Int-C(15.5mg,37.3μmol)、Pd(dppf)Cl2(2.7mg,3.7μmol)和KOAc(10.9mg,111μmol)加入到1,4-二氧六环(0.5mL)和H2O(0.5mL)中，氮气保护后80℃反应8hr。反应完成后降至室温，浓缩，粗品经prep-TLC(流动相PE/EA＝1:2)纯化，得到化合物43c(10.0mg)。MS m/z(ESI)：510.1[M-55]+。
第三步：2-(4-(4-(氨基甲基)-1-氧代-1,2,3,4-四氢异喹啉-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-环丙氧基-3-氟苄腈(43)的合成
在冰浴下将化合物43c(10.0mg,17.6μmol)加入至HCl/EtOAc(4M,2.0mL)中，然后升温至25℃反应1hr。减压浓缩，得到化合物43(9.33mg)。MS m/z(ESI)：466.0[M+H]+。
1H 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),3.24-3.17(m,1H),3.09-2.99(m,1H),2.97-2.82(m,1H),0.94-0.78(m,4H).
实施例50：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-(1-甲基环丙氧基)苄腈(50/51)
第一步：4-氯-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)-6-(1-甲基环丙氧基)苄腈(20b)的合成
冰浴下向反应瓶中加入化合物20a(68.3mg,948μmol)和THF(3mL)，后缓慢加入NaH(47.4mg,1.19mmol)并搅拌30min，最后滴入Int-B(300mg,790μmol)的THF(1mL)溶液，加完后50℃反应2hr。反应完毕降温到0℃，加入5mL水淬灭，然后用乙酸乙酯萃取，合并有机相并减压浓缩，粗品经硅胶柱层析(流动相PE:EA＝3:1)纯化，得到化合物20b(200.0mg)。MS m/z(ESI)：431.8[M+H]+。
第二步：((7-(5-(3-氯-6-氰基-2-氟-5-(1-甲基环丙氧基)苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(20c)的合成
将化合物20b(200.0mg,463μmol)、Int-A(223.0mg,556μmol)、CsF(211.0mg,1.39mmol)和cataCXiumAPdG3(33.8mg,46.3μmol)加入到1,4-二氧六环(3mL)和H2O(0.6mL)中，氮气保护后60℃ 反应12hr。反应完成后降至室温，减压浓缩，粗品经Prep-TLC(展开剂EA：PE＝1:1)纯化，得化合物20c。化合物20c经过SFC拆分(柱子:DAICEL CHIRALPAK IC(250mm*30mm,10um)；流动相:[Neu-ETOH]；B％:40％-40％,A4.4；60min)，得到化合物20c峰1(45.0mg)，MS m/z(ESI)：579.1[M+H]+；和化合物20c峰2(40.0mg)，MS m/z(ESI)：579.1[M+H]+。
第三步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-(1-甲基环丙氧基)苄腈(50/51)的合成
将化合物20c峰1(45.0mg,77.7μmol)加入到HCl的EA溶液(3mL)中，25℃反应0.5hr。反应结束后将反应液浓缩干，经过pre-HPLC(柱子:Welch Xtimate C18 150*25mm*5um；流动相:[water(HCl)-ACN]；B％:12％-42％,8min)得到化合物50(29.8mg)。MS m/z(ESI)：479.1[M+H]+。
1H 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).
将化合物20c峰2(40.0mg,69.1μmol)加入到HCl的EA溶液(2mL)中，25℃反应0.5hr。反应结束后将反应液浓缩干，经过pre-HPLC(柱子:Welch Xtimate C18 150*25mm*5um；流动相:[water(HCl)-ACN]；B％:11％-41％,8min)得到化合物51(24.0mg)。MS m/z(ESI)：479.1[M+H]+。
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).
实施例52：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-(3-甲基氮杂环丁烷-1-基)苄腈(52)
第一步：4-氯-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)-6-(3-甲基氮杂环丁烷-1-基)苄腈(52b)合成
将化合物52a(17.0mg,158μmol)和K2CO3(65.6mg,474μmol)加入到DMF(1.0mL)中，并在25℃搅拌10分钟，然后加入Int-B(60.0mg,158μmol)，反应加热至80℃搅拌12hr。反应完成后降至室温，加水淬灭并用乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，过滤后减压浓缩，粗品经硅胶柱层析(流动相PE:EA＝3:1)纯化，得化合物52b(40.0mg)。MS m/z(ESI)：430.9[M+H]+
第二步：((7-(5-(3-氯-6-氰基-2-氟-5-(3-甲基氮杂环丁烷-1-基)苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)甲酸叔丁酯(52c)的合成
将化合物52b(40.0mg,92.9μmol)、Int-A(52.2mg,130μmol)、CsF(42.3mg,278μmol)和cataCXiumAPdG3(6.7mg,9.3μmol)加入到1,4-二氧六环(1.0mL)和H2O(0.2mL)中，氮气保护后60℃反应12hr。反应完成后降至室温，过滤后减压浓缩，粗品经Prep-TLC(展开剂EA：PE＝1:1)纯化，得化合物52c(20.0mg)。MS m/z(ESI)：578.2[M+H]+。
第三步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-(3-甲基氮杂环丁烷-1-基)苄腈(52)的合成
将化合物52c(20.0mg,34.6μmol)加入至二氯甲烷(2.0mL)中，降温至0℃后滴加TFA(78.9mg,692.0μmol,0.05mL)，然后25℃反应1hr。反应完毕后减压浓缩除去溶剂，粗产物经Pre-HPLC(柱子:Phenomenex C18 150*25mm*10um；流动相:[水(NH4HCO3)-ACN]；B％:24％-54％,8min)纯化，得到化合物52(7.95mg)。MS m/z(ESI)：478.1[M+H]+。
1H 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).
以下化合物通过实施例52所述方法和一般步骤制备得到，所需其它原料可通过商业购买，或由有机合成领域有经验的合成人员从商业购买试剂使用常规反应合成获得。



实施例80：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-((6-(4-氟-1H-吡唑-1-基)吡啶-3-基)氧基)苄腈(80)
第一步：5-苄氧基-2-氟吡啶(化合物80-2)的合成
将化合物80-1(500mg，4.42mmol)、K2CO3(1.22g，8.84mmol)溶于干燥DMF(10mL)中，随后加入苄溴(1.13g,6.63mmol)，反应体系于20℃反应12hr。反应完成后加水稀释，乙酸乙酯萃取，有机层干燥浓缩，粗品经硅胶柱层析纯化(流动相PE：EA＝92:8)得化合物80-2(885mg)。MS(ESI,m/z):204.1[M+H]+。
第二步：5-苄氧基-2-(4-氟-1H-吡唑-1-基)吡啶(化合物80-4)的合成
反应瓶中加入80-2(850mg,4.18mmol)、80-3(540.01mg,6.27mmol)、Cs2CO3(2.73g 8.37mmol)、DMF(10mL)，氮气保护后120℃反应8hr。反应完成降至室温，加水稀释，乙酸乙酯萃取，有机层干燥浓缩，粗品经硅胶柱层析纯化(流动相PE：EA＝90:10)得化合物80-4(910mg)。MS(ESI,m/z):270.1[M+H]+。
第三步：6-(4-氟-1H-吡唑-1-基)吡啶-3-醇(化合物80-5)的合成
将化合物80-4(890mg,3.31mmol)溶解于MeOH(15mL)中，加入Pd/C(401.42mg,3.31mmol)，氢气球氢气置换后于25℃反应2hr。反应完成后，过滤，滤液减压浓缩得化合物80-5(560mg)。MS(ESI,m/z):180.1[M+H]+。
第四步：((7-(5-(3-氯-6-氰基-2,5-二氟苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(化合物80-6)的合成
反应瓶中加入Int-B(30mg,79.04μmol)、Int-A(31.72mg,79.04μmol)、Pd(dppf)Cl2.DCM(6.41mg,7.90μmol)、NaHCO3(19.92mg,237.13μmol)及二氧六环(5mL)与H2O(0.5mL)，氮气保护后80℃反应5hr。反应完成后降至室温减压浓缩，粗品经Prep-TLC分离纯化(流动相DCM:MeOH＝20:1)得化合物80-6(10mg)。MS(ESI,m/z):527.1[M+H]+。
第五步：((7-(5-(3-氯-6-氰基-2-氟-5-((6-(4-氟-1H-吡唑-1-基)吡啶-3-基)氧基)苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(化合物80-7)的合成
将化合物80-5(5.10mg，28.47μmol)溶于无水THF(5mL)中，缓慢加入NaH(1.14mg，28.47μmol)搅拌30min，加入化合物80-6(10mg,18.98μmol)，N2保护后升温至60℃反应2hr。反应完成降至室温，加入适量甲醇淬灭反应，减压浓缩，经Prep-TLC分离纯化(流动相DCM:MeOH＝20:1)得化合物80-7(6mg)。MS(ESI,m/z):686.1[M+H]+。
第六步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-3-氟-6-((6-(4-氟-1H-吡唑-1-基)吡啶-3-基)氧基)苄腈(化合物80)的合成
将化合物80-7(6mg,8.75μmol)加入至二氯甲烷(3mL)中，滴加TFA(0.5mL)，加毕于25℃反应2hr。反应完毕后加入三乙胺调至弱碱性，减压浓缩，粗产物经prep-HPLC纯化，得到化合物80(1.5mg)。MS(ESI,m/z):586.2[M+H]+。
1H 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)。
实施例81：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-(1-(二氟甲基)环丙氧基)-3-氟苄腈(81和82,83)

第一步：4-氯-3,6-二氟-2-碘苄腈(化合物Int-Bb)的合成
将化合物Int-Ba(70.0g,403mmol)溶于DCE(1.80L)中，然后加入Pd(OAc)2(9.06g,40.3mmol)NIS(227g,1.01mol)和TFA(46.0g,403mmol,30.0mL)。反应加热至75℃持续搅拌12小时。按照以上步骤平行两个批次反应。TLC(石油醚/乙酸乙酯＝10:1,Rf＝0.5)检测原料完全转化。反应液中加入饱和食盐水(2.00L)淬灭，然后使用二氯甲烷(2.00L x 3)萃取。合并有机相后用无水硫酸钠干燥、抽滤、浓缩得到粗产物，经过硅胶柱层析(流动相:石油醚/乙酸乙酯＝100:1到10:1)分离纯化，浓缩后得到化合物Int-Bb(184g)。
第二步：4-氯-3,6-二氟-2-(1-甲基-1H-吡唑-5-基)苄腈(化合物Int-Bc)的合成
将Int-Bb(60.0g,188mmol)、1-甲基-1H-吡唑-5-硼酸频哪醇酯(58.8g,283mmol)溶于1,4-二氧六环(2.80L)和H2O(280mL)中，然后加入Pd(dbtpf)Cl2(12.3g,18.8mmol)和DIEA(122g,942mmol,164mL)。置换氮气后反应温度升高至80℃搅拌1小时。LCMS监测Int-Bb完全转化。降温至室温后，减压蒸馏浓缩除去溶剂，然后用饱和食盐水(3.00L)和乙酸乙酯(3.00L x 3)萃取，合并有机相，无水硫酸钠干燥，抽滤，浓缩，经硅胶柱层析(流动相:石油醚/乙酸乙酯＝100:1到4:1)分离纯化，浓缩得到化合物Int-Bc(25.1g)。MS m/z(ESI):254.0[M+H]+。
第三步：1-(5-氯-2-氰基-4-氟-3-(1-甲基-1H-吡唑-5-基)苯氧基)环丙烷-1-甲酸甲酯(化合物81-2)的合成
将化合物Int-Bc(25.1g,99.0mmol),81-1(23.0g,198mmol)溶于THF(500mL)中，在冰浴下缓慢加入NaH(8.71g,218mmol)。然后反应缓慢升温至室温继续搅拌3小时。LCMS监测反应完全转化。反应在0℃下用氯化铵水溶液(500mL)淬灭，然后用乙酸乙酯(500mL x 3)萃取，合并有机相，用无水硫酸钠干燥，抽滤，浓缩后经硅胶柱层析(流动相:石油醚/乙酸乙酯＝100:1到3:1)分离纯化，浓缩得到化合物81-2(29.5g)。MS m/z(ESI):350.1[M+H]+。
第四步：4-氯-3-氟-6-(1-甲酰基环丙氧基)-2-(1-甲基-1H-吡唑-5-基)苄腈(化合物81-3)的合成
制备LDBBA:将叔丁醇(5.26mL,55mmol)溶于THF(25mL)中，降温至0℃后，缓慢滴加n-BuLi(22mL,2.5M in hexane,55mmol)并升温至10℃搅拌1小时。然后降温到0℃后滴加DIBALH(50mL,1.0M in hexane,50mmol)，滴加完成后保持在10℃继续搅拌2小时，得到LDBBA的溶液。
在-65℃条件下，将上述LDBBA溶液缓慢加入到化合物81-2(29.5g,84.4mmol)的THF(300mL)溶液中，并保持在-65℃反应1小时。LCMS监测反应完全。在冰浴下加入饱和氯化铵水溶液淬灭反应，并用乙酸乙酯(300mL x 3)萃取，合并有机相，无水硫酸钠干燥，抽滤，浓缩，经硅胶柱层析(流动相:石油醚/乙酸乙酯＝100:1到1:1)分离纯化，浓缩后得到化合物81-3(24.7g)。MS m/z(ESI):320.2[M+H]+。
第五步：4-氯-6-(1-(二氟甲基)环丙氧基)-3-氟-2-(1-甲基-1H-吡唑-5-基)苄腈(化合物81-4)的合成
在冰浴下，将DAST(29.9g,185mmol,24.5mL)缓慢加入到化合物81-3(23.7g,74.1mmol)的 DCM(240mL)溶液中，并保持在0℃反应12小时。LCMS监测反应完全。然后相反应液中加H2O(200mL)淬灭，然后用二氯甲烷(200mL x 3)萃取，合并有机相，无水硫酸钠干燥，抽滤，浓缩得到化合物81-4(27.0g)。MS m/z(ESI):342.2[M+H]+。
第六步：4-氯-6-(1-(二氟甲基)环丙氧基)-3-氟-2-(4-碘-1-甲基-1H-吡唑-5-基)苄腈(化合物81-5)的合成
将81-4(27.0g,79.0mmol)溶于HOAc(400mL)中，然后加入NIS(26.7g,119mmol)。反应升温至80℃反应12小时。LCMS监测原料反应完全，反应液直接减压蒸馏浓缩后经过硅胶柱层析(流动相:石油醚/乙酸乙酯＝100:1到3:1)分离，纯化、浓缩后得到化合物81-5(29.8g)。MS m/z(ESI):468.0[M+H]+。
第七步：((7-(5-(3-氯-6-氰基-5-(1-(二氟甲基)环丙氧基)-2-氟苯基)-1-甲基-1H-吡唑-4-基)-4-氧代-3,4-二氢酞嗪-1-基)甲基)氨基甲酸叔丁酯(化合物81-6)的合成
将化合物81-5(8.00g,17.1mmol)、Int-A(10.3g,25.7mmol)溶于1,4-二氧六环(160mL)和H2O(32mL)中，然后在氮气氛围下加入CsF(7.80g)和cataCXiumA Pd G3(1.25g,1.71mmol)。反应升温至100℃搅拌12小时。LCMS监测到产物生成。浓缩后加H2O(50mL)，然后用乙酸乙酯(100mL x 3)萃取，合并有机相，无水硫酸钠干燥，抽滤，经硅胶柱层析(流动相:石油醚/乙酸乙酯＝100:1到1:1)分离纯化，浓缩后得到化合物81-6(3.87g)。MS m/z(ESI):615.3[M+H]+。
化合物81-6经prep-SFC(手性柱:DAICEL CHIRALPAK IC(250mm*30mm,10um)；流动相:[CO2-iPrOH(0.1％NH3H2O)]；B％:30％,isocratic elution mode)拆分得到化合物83-1(125mg,保留时间＝3.611min)，MS m/z(ESI):615.2[M+H]+和82-1(117mg,保留时间＝4.087min)，MS m/z(ESI):615.2[M+H]+。
第八步：2-(4-(4-(氨基甲基)-1-氧代-1,2-二氢酞嗪-6-基)-1-甲基-1H-吡唑-5-基)-4-氯-6-(1-(二氟甲基)环丙氧基)-3-氟苄腈(化合物81和82,83)的合成
将化合物81-6(42mg,68.29μmol)溶于CH2Cl2(2mL)中，然后加入TFA(1mL)。反应在25℃搅拌2小时。LCMS监测反应完全。经prep-HPLC分离纯化，冻干得化合物81的三氟醋酸盐(15.89mg)。MS(ESI,m/z):515.1[M+H]+。
1H 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)。
将化合物82-1(110mg,179μmol)溶于CH2Cl2(3mL)，然后加入TFA(0.6mL)。反应在25℃搅拌1小时。LCMS监测反应完全。直接浓缩，经prep-HPLC分离纯化得到化合物82(15mg)。MS m/z(ESI):515.3[M+H]+。
1H 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)。
将化合物83-1(117mg,190μmol)溶于CH2Cl2(3mL)，然后加入TFA(0.6mL)。反应在25℃搅拌1小时。LCMS监测反应完全。直接浓缩，经prep-HPLC分离纯化得到化合物83(32mg)。MS m/z(ESI):515.3[M+H]+。
1H 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)。
以下化合物通过实施例81所述方法和一般步骤制备得到，所需其它原料可通过商业购买，或由有机合成领域有经验的合成人员从商业购买试剂使用常规反应合成获得。
分离方法
实施例中某些化合物的Prep-HPLC纯化采用Aglient 1260型或Waters 2489型HPLC进行，分离柱型号为Waters SunFire Prep C18OBD(19mm×150mm×5.0μm)、Waters Xbridge Prep C18OBD(19mm×150mm×5.0μm)或YMC Actus Triart C18(20mm×150mm×5.0μm)，柱温均为25℃，检测波长为214nm、254nm或280nm，流动相A为乙腈，流动相B为0.05％甲酸水溶液或者0.05％碳酸氢铵水溶液或者0.05％TFA水溶液，流动相的体积比根据化合物的极性不同而进行调节；流动相流速为28mL/min。
生物学评价
实验例1：化合物对PRMT5-MTA甲基转移酶活性抑制作用测试
实验方法一：将配置好的蛋白溶液(PRMT5/MEP50和MTA混合液)分别与不同浓度的测试化合物(1000nM起始，5倍稀释，7个点)25℃预孵育30min后，加入配置好的底物溶液(Biotinylated histone H4peptide)，25℃孵育反应90min；反应结束后，加入配好的检测试剂混合液(Protein A-Eu、Anti-Histone H4antibody和Streptavidin-D2)，25℃孵育60min，使用BMG酶标仪检测荧光信号比值(Ratio)。
以溶剂组(DMSO)为阴性对照、反应缓冲液组(不含PRMT5·MTA酶)为空白对照，按照下式计算不同浓度化合物的百分比抑制率：
百分比抑制率＝(阴性对照Ratio-化合物Ratio)/(阴性对照Ratio-空白对照Ratio)×100％；
使用GraphPad Prism 8中“log(inhibitor)vs.response--Variable slope”四参数方程Y＝Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*Hillslope))对检测信号值进行拟合，计算IC50值。其中Y为相对抑制活性百分比，Top和Bottom分别为拟合曲线的最大值与最小值，X为化合物的对数浓度，Hillslope为曲线斜率。按照上述方法测定化合物对PRMT5-MTA甲基转移酶的抑制作用，结果如表1中所示。
表1.本发明的化合物对PRMT5-MTA的抑制活性

实验方法二：根据PRMT5TR-FRET试剂盒(BPS Bioscience)的说明测定本发明化合物对PRMT5-MTA甲基转移酶活性抑制作用。将PRMT5/MEP50酶和MTA混合物分别与不同浓度的测试化合物(100nM、20nM、4nM)在25℃下预孵育30min后，加入Biotinylated histone H4 peptide/S-adenosylmethionine混合工作液，25℃孵育反应120min；加入Eu-labeled antibody工作液，25℃缓慢震荡30min，再加入Dye-labeled acceptor工作液，25℃缓慢震荡30min，酶标仪检测荧光信号比值(Ratio)。
以溶媒组(DMSO)为阴性对照，缓冲液组(不含PRMT5·MTA酶)为空白对照，按照下式计算不同浓度化合物的百分比抑制率：
百分比抑制率＝(阴性对照Ratio-化合物Ratio)/(阴性对照Ratio-空白对照Ratio)×100％；
当百分比抑制率介于30-80％时，按照下述公式计算化合物的半数抑制浓度(IC50)或范围：
IC50＝X×(1-百分比抑制率(％))/百分比抑制率(％)，其中：X为抑制率介于30-80％时化合物的测试浓度。
按照上述方法测定化合物对PRMT5-MTA的抑制作用，结果如表2中所示。
表2.本发明的化合物对PRMT5-MTA的抑制活性
实验结果表明，本发明化合物对PRMT5-MTA酶均有较强的抑制作用。
实验例2：化合物对MTAP Deleted/Parental HCT116细胞增殖抑制试验
实验方法一：通过测试本发明的化合物对癌细胞生长的影响，进一步评估本发明的化合物对癌细胞增殖的抑制作用。本实施例中选用了MTAP Deleted/Parental HCT116细胞，来自Pharmaron Inc.。
将MTAP Deleted/Parental HCT116细胞体外单层培养，培养条件为10％FBS+1％P/S的MCCOYS5A培养基，37℃,5％CO2。消化对数生长期细胞并调整浓度，每孔150个接种于384孔板培养过夜，加入预先稀释的化合物(10000nM起始，4倍稀释，10个点或5000nM起始，5倍稀释，8个点)，阴性对照组加DMSO，空白对照组加培养基，于37℃，5％CO2孵箱中培养10天后，每孔加入40μL CellTiter-Glo,在酶标仪化学发光检测模式下读取相对化学发光单位值。
按照下式计算不同浓度化合物的百分比抑制率：
百分比抑制率＝(1-(测试化合物的化学发光信号值-空白对照的化学发光信号值)/(阴性对照的化学发光信号值-空白对照的化学发光信号值))×100％
将不同浓度化合物的百分比抑制率相对于化合物浓度作图，按照四参数模型拟合曲线，通过下式计算IC50值：
y＝Min+(Max-Min)/(1+(x/IC50)^(-Hillslope))，其中：y为百分比抑制率；Max和Min分别为拟合曲线的最大值与最小值；x为化合物的对数浓度；且Hillslope为曲线斜率。
按照上述方法测定化合物对MTAP Deleted/Parental HCT116细胞的增殖抑制活性，结果如表3所示。
表3.本发明的化合物对MTAP Deleted/Parental HCT116细胞的增殖抑制活性
实验方法二：本实施例中选用了MTAP Deleted/Parental HCT116细胞，购自HORIZON。
将MTAP Deleted/Parental HCT116细胞体外单层培养，培养条件为10％FBS+1％P/S的RPMI6140培养基，37℃、5％CO2。消化对数生长期细胞并调整浓度，每孔250个接种于96孔板培养过夜，加入预先稀释的化合物(5000nM起始，4倍稀释，9个点)，阴性对照组加DMSO，空白对照组加培养基，于37℃，5％CO2孵箱中培养8天后，每孔加入50μl CellTiter-Glo室温避光裂解10分钟，将待测液转至96孔不透光白板中，然后在酶标仪化学发光检测模式下读取相对化学发光单位值。
按照下式计算不同浓度化合物的百分比抑制率：
百分比抑制率＝(1-(测试化合物的化学发光信号值-空白对照的化学发光信号值)/(阴性对照的化学发光信号值-空白对照的化学发光信号值))×100％
将不同浓度化合物的百分比抑制率相对于化合物浓度作图，按照四参数模型拟合曲线，通过下式计算IC50值：
y＝Min+(Max-Min)/(1+(x/IC50)^(-Hillslope))，其中：y为百分比抑制率；Max和Min分别为拟合曲线的最大值与最小值；x为化合物的对数浓度；且Hillslope为曲线斜率。
按照上述方法测定化合物对MTAP Deleted/Parental HCT116细胞的增殖抑制活性，结果如表4所示。
表4.本发明的化合物对MTAP Deleted/Parental HCT116细胞的增殖抑制活性

实验结果表明，本发明化合物对MTAP Deleted的HCT116细胞有较强的抑制作用，且对MTAP Parental的HCT116细胞具有一定的选择性。
实验例3：化合物在balb/c小鼠体内药代动力学测试
通过灌胃(PO)向雌性Balb/c小鼠给予本发明的化合物，考察药代动力学特点。
将本发明的化合物51和MRTX-1719(根据专利WO2021050915A1实施例16-8制备)进行PO给药，给药剂量是30mg/kg，给药溶媒为0.5％MC+0.2％Tween 80水溶液。在给药前(0h)以及给药后0.25、0.5、1、2、4、6、8、10、24h采集血样；经眼眶取血40-50μl，置于K2-EDTA抗凝管中，4000rpm离心10min(4℃)，分离血浆，-80℃保存。取10μl血浆样品加入90μl含内标的乙腈，振摇混匀后，于4000rpm，4℃下离心10min，取上清液进行LC-MS/MS分析。应用WinNonlin 6.3软件，采用非房室模型计算药代动力学参数，结果见表5。
表5：化合物在小鼠体内的药代动力学参数
结论：
本发明的化合物51通过PO给药后在小鼠体内暴露量高，血药浓度高。
除本文中描述的那些外，根据前述描述，本发明的多种修改对本领域技术人员而言会是显而易见的。这样的修改也意图落入所附权利要求书的范围内。本申请中所引用的各参考文献(包括所有专利、专利申请、期刊文章、书籍及任何其它公开)均以其整体援引加入本文。

Claims (12)

  1. 化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，所述化合物具有式(I)的结构：
    其中：
    R1为
    R2和R6各自独立地选自H、卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、C3-8环烷氧基或3-8元杂环氧基；
    R3、R4和R5各自独立地选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
    -X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
    -X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
    -X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
    -X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
    或者R3与R4连同其所连接的原子一起形成4-11元螺杂环基，
    或者R4与R5连同其所连接的原子一起形成4-11元螺杂环基，
    条件是，当R1为时，R3、R4和R5至少有一个选自上述-X-L-C3-8环烷基、-X-L-3-8元杂环烷基、-X-L-C6-10芳基、-X-L-5-10元杂芳基、-X-L-5-10元桥环基、-X-L-5-10元螺环基、3-8元杂环基；
    X在每次出现时各自独立地选自O、S或NR7；
    L在每次出现时各自独立地为直接键或-(CH2)n-；
    n为1或2；
    R7选自H、C1-4烷基或C1-4卤代烷基；
    R8和R9独立地选自H、C1-4烷基或C1-4卤代烷基。
  2. 权利要求1所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，其中，R2和R6各自独立地选自H、卤素、CN、C1-4烷基、C1-4卤代烷基或3-8元杂环氧基；
    优选地，R2为CN；R6为F、Cl、Br或I。
  3. 权利要求1或2所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，其中所述化合物具有式(I-A)的结构：
    其中：
    R3、R4和R5如权利要求1所定义。
  4. 权利要求1-3任一项所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，其中，
    R3选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
    -X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
    -X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基、取代或未取代的5-10元杂芳基取代，
    -X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
    -X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代；
    优选地，R3选自H、3-6元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
    -X-L-C3-6环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-3-6元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、C1-4卤代烷基、取代或未取代的5-10元杂芳基取代，
    -X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个取代或未取代的5-10元杂芳基取代，
    -X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
    -X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代；
    优选地，R3选自H、3-6元杂环基，所述杂环基被一个或多个卤素、CN、羟基、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基或C1-4羟烷基取代，
    -X-L-C3-6环烷基，所述环烷基被一个或多个卤素、CN、羟基、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-3-6元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-C6-10芳基，所述芳基任选地被一个或多个卤素、C1-4卤代烷基、取代或未取代的5-10元杂芳基取代，
    -X-L-5-10元杂芳基，所述杂芳基任选地被一个或多个取代或未取代的5-10元杂芳基取代，
    -X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
    -X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代；
    X在每次出现时各自独立地选自O或NR7；
    L在每次出现时各自独立地选自直接键或-(CH2)-；
    R7选自H或C1-4烷基；
    R8和R9独立地选自H、C1-4烷基或C1-4卤代烷基；
    优选地，R3选自H、氮杂环丁烷、2-氮杂螺[3.3]庚烷，所述的氮杂环丁烷被一个或多个F、CN、CF3、甲基、甲氧基、乙氧基、CHF2或N(CH3)2取代；
    -X-L-C3-6环烷基，所述的环烷基选自环丙基、环丁基、环戊基或环已基，所述环丙基、环丁基、环戊基或环已基被一个或多个F、羟基、甲基、CHF2或CF3取代；
    -X-L-3-6元杂环基，所述的杂环基选自氧杂环丁基、氮杂环丁烷、四氢呋喃、四氢吡喃、哌啶或吡咯烷，所述氧杂环丁基、氮杂环丁烷、四氢呋喃、四氢吡喃、哌啶、吡咯烷任选地被一个或多个F、-C(O)CH3、甲基、乙基、异丙基或叔丁基取代；
    -X-L-C6-10芳基，所述芳基为苯基，所述苯基任选地被一个或多个F、CF3、取代，
    -X-L-5-10元杂芳基，所述杂芳基为吡啶基，所述吡啶基任选地被一个或多个取代，
    -X-L-5-10元桥环基，所述桥环为双环[1.1.1]戊烷，
    -X-L-5-10元螺环基，所述的螺环基选自7-氮杂螺环[3.5]壬烷、螺[3.3]庚烷或2-氧杂螺[3.3]庚烷，所述7-氮杂螺环[3.5]壬烷、螺[3.3]庚烷或2-氧杂螺[3.3]庚烷任选地被一个或多个甲基取代；
    X在每次出现时各自独立地选自O、NH或N(CH3)；
    L在每次出现时各自独立地选自直接键或-(CH2)-；
    优选地，R3选自
  5. 权利要求1-4任一项所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，其中，
    R4选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代，
    -X-L-C3-8环烷基，所述环烷基被一个或多个卤素、CN、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-3-8元杂环基，所述杂环基任选地被一个或多个卤素、CN、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4杂烷基取代，
    -X-L-5-10元桥环基，所述桥环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代，
    -X-L-5-10元螺环基，所述螺环基任选地被一个或多个卤素、C1-4烷基、C1-4卤代烷基取代；
    优选地，R4选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代；
    优选地，R4选自H、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代；
    优选地，R4选自H或
  6. 权利要求1-5任一项所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，其中，
    R5选自H、卤素、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、C1-4羟烷基、C3-8环烷基、3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代；
    优选地，R5选自H、C1-4烷基、卤素或3-8元杂环基，所述杂环基被一个或多个卤素、CN、-NR8R9、-COCH3、C1-4烷基、C1-4卤代烷基、C1-4烷氧基、C1-4卤代烷氧基、3-6元杂环基取代；
    优选地，R5选自H、甲基、F、Cl、Br或I。
  7. 权利要求1-6任一项所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，其中，所述化合物选自：



  8. 药物组合物，其包含预防或治疗有效量的权利要求1-7任一项所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物以及一种或多种药学上可接受的载体。
  9. 权利要求1-7任一项所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物或者权利要求8所述药物组合物在制备用于预防或治疗与PRMT5活性相关的疾病或病况的药物中的用途；
    优选地，所述与PRMT5活性相关的疾病或病况为MTAP缺失的癌症或肿瘤；更优选地，所述癌症或肿瘤优选为食管癌、肺癌、胰腺癌、胶质母细胞瘤、胆管癌、膀胱癌、间皮瘤、结直肠癌、头颈部鳞状细胞癌、淋巴瘤。
  10. 一种预防或治疗与PRMT5活性相关的疾病或病况的方法，所述方法包括向需要其的个体给药有效量的权利要求1-7任一项所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物，或者权利要求8所述的药物组合物；
    优选地，所述与PRMT5活性相关的疾病或病况为MTAP缺失的癌症或肿瘤；更优选地，所述癌症或肿瘤优选为食管癌、肺癌、胰腺癌、胶质母细胞瘤、胆管癌、膀胱癌、间皮瘤、结直肠癌、头颈部鳞状细胞癌、淋巴瘤。
  11. 权利要求1-7任一项所述的化合物或其药学上可接受的盐、立体异构体、互变异构体或同位素标记的化合物或者权利要求8所述药物组合物，其用于预防或治疗与PRMT5活性相关的疾病或病况；
    优选地，所述与PRMT5活性相关的疾病或病况为MTAP缺失的癌症或肿瘤；更优选地，所述 癌症或肿瘤优选为食管癌、肺癌、胰腺癌、胶质母细胞瘤、胆管癌、膀胱癌、间皮瘤、结直肠癌、头颈部鳞状细胞癌、淋巴瘤。
  12. 权利要求3所述化合物的制备方法，其包括以下步骤：
    其中：
    PG为氨基保护基，优选为叔丁氧羰基；
    其余各基团如权利要求3所定义；
    第一步：化合物I-A-1和I-A-2经取代反应生成化合物I-A-3；
    第二步：化合物I-A-3和I-A-4经Suzuki偶联反应生成化合物I-A-5；
    第三步：化合物I-A-5在酸性条件下脱除保护基生成式I-A的化合物。
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