WO2020156479A1 - Cyclopropene- and benzofuran-substituted azaaryl compound, and intermediate, preparation method and application thereof - Google Patents

Abstract

A cyclopropene- and benzofuran-substituted azaaryl compound, and an intermediate, preparation method and application thereof. Provided are a method for preparing such a compound, a composition containing such a compound, and a use of such a compound in the preparation of a drug for treating a disease or disorder related to an EED-protein- and/or PRC2-protein-complex action mechanism.

Description

Cyclopropenacbenzofuran substituted azaaryl compound, its intermediate, preparation method and application

This application claims the priority of the Chinese patent application 201910106971.7 whose filing date is 2019/2/2. This application quotes the full text of the aforementioned Chinese patent application.

Technical field

The present invention relates to a novel cyclopropenacbenzofuran substituted azaaryl compound, its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer or its solvate and their isotope label . The present invention also provides a preparation method of the compound, a composition containing the compound, and a pharmaceutical application for treating diseases related to the mechanism of action of the EED protein and/or PRC2 protein complex.

Background technique

PcG (Polycomb Group) protein is an important class of chromatin modifying enzymes. It modifies chromatin to regulate gene transcription, which plays an important role in the growth, differentiation and long-term cellular memory of stem cells. In mammalian cells, PcG proteins are mainly divided into two types of transcription repressive complexes, namely PRC1 (Polycomb Repressive Complex1) and PRC2 (Polycomb Repressive Complex2). Among them, PRC2 inhibits the expression of related genes by methylating the lysine 27 (H3K27) of histone 3 in the chromatin. The PRC2 protein complex is mainly composed of core proteins such as EZH2 (Enhancer of Zeste Homolog2) (or its very similar homologous protein EZH1), EED (Embryonic ectoderm Development) and SUZ12 (Suppressor of Zeste12). Among them, EZH2 has enzymatic catalytic activity. The SET (Su(var), E(Z), and Trithorax) protein domain can transfer the methyl group of the substrate SAM (S-adenosyl-L-methionine) to H3K27, thereby One to three methylation modification of H3K27. The catalytic activity of EZH2 also depends on other components of PRC2, such as the EED protein that belongs to the WD40 repeat structure protein family. The combination of EED and trimethylated H3K27Me3 on the one hand has a great allosteric promotion effect on the enzyme catalytic function of EZH2, on the other hand, it can also locate the PCR2 complex on the chromatin that needs to be modified. The functional abnormalities of PRC2, such as overexpression or gain-of-function mutations of EZH2, are associated with many clinical tumor diseases, including lung cancer, breast cancer, rectal cancer, prostate cancer, bladder cancer, pancreatic cancer, sarcoma, and lymphoma. PRC2 is also related to a variety of cellular immune functions. For example, EZH2 is involved in regulating lymphocyte activation and can also work with glycolysis to promote T cell responses to tumor cells. Therefore, the development of small molecule inhibitors of PRC2 has important and broad drug development value.

The research and development surrounding PRC2 inhibitors is mainly to develop two strategies, EZH2 inhibitors and EED inhibitors. EZH2 inhibitors currently entering the clinic include EPZ-6438 (Epizyme, clinical phase II), GSK2816126 (GSK, clinical phase I), and CPI-1205 (Constellation, clinical phase I) and so on. Although many EZH2 inhibitors have entered the clinical research phase, these inhibitors all contain a common 2-pyridone pharmacophore. Moreover, in the use of existing EZH2 inhibitors in clinical treatment, secondary mutations have begun to appear. EED inhibitors have an allosteric inhibitory effect on EZH2 enzyme function, and can achieve the same or similar biological functions as EZH2. Moreover, on the one hand, EED inhibitors can overcome the problem of EZH2 resistance. On the other hand, EED inhibitors can be combined with EZH2 inhibitors to achieve better synergistic effects. Therefore, the development of new EED inhibitors is very important Meaning.

Summary of the invention

The cyclopropenyl benzofuran substituted azaaryl compound provided by the present invention is a new type of EED inhibitor, exhibits good inhibitory activity on tumor cells, and has broad prospects for drug development.

In the first aspect, the present invention provides a compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer or its solvate:

among them,

X is independently C or N;

R ^{2 is} independently H or halogen (e.g. fluorine);

R ^{3 is} independently H or halogen (e.g. fluorine);

R ^{4 is} independently H or halogen (e.g. fluorine);

n is independently 0 or 1;

When X=C, then n is 1, forming a pyridopyrimidine structure

When X=N, then n is 0, forming a triazolopyrimidine structure

R ^1c取代或未取代的烯基(所述烯基例如C _2-10烯基，优选C _2-6烯基，更优选乙烯基)、R ^1b取代或未取代的C _5-8环烯基(所述C _5-8环烯基例如

R ^1b取代或未取代的C _5-8杂环烯基(所述C _5-8杂环烯基优选为具有1至3个独立选自氮、氧或硫的杂原子的C _5-8杂环烯基，例如

或

)、0-3个R ^1d取代的C _6-10芳基(所述C _6-10芳基例如苯基)或0-3个R ^1d取代的“具有C _1-20碳原子及1-4个独立选自N、NR ^1de1、O或S(O) _0-2的杂原子的杂芳基”(所述C _1-20碳原子优选为C _1-10碳原子，所述杂芳基优选为5-12元杂芳基，例如

R ^{1 is} independently hydrogen, halogen (for example, fluorine, bromine or iodine), cyano, R ^1a substituted or unsubstituted C _1-8 alkyl (the C _1-8 alkyl such as C _1-4 alkyl, It can be methyl, ethyl, n-propyl or isopropyl), C _1-8 haloalkyl (e.g. C _1-4 haloalkyl), R ^1a substituted or unsubstituted C _3-8 cycloalkyl (the C _3-8 cycloalkyl such as cyclohexyl), R ^1b substituted or unsubstituted C _3-8 cycloalkyl (the C _3-8 cycloalkyl such as cyclohexyl), R ^1b substituted or unsubstituted C _{3 -8} heteroalkyl (said C _3-8 heteroalkyl having preferably C 1 to 3 hetero atoms independently selected from nitrogen, oxygen, sulfur, or _3-8 heteroalkyl, e.g.

R ^1c substituted or unsubstituted alkenyl (the alkenyl is for example C _2-10 alkenyl, preferably C _2-6 alkenyl, more preferably vinyl), R ^1b substituted or unsubstituted C _5-8 cycloalkenyl (The C _5-8 cycloalkenyl is for example

R ^1b substituted or unsubstituted heterocyclyl C _5-8 alkenyl group (said C _5-8 heterocycloalkenyl group having preferably C 1 to 3 hetero atoms independently selected from nitrogen, oxygen, or sulfur heteroatoms _5-8 Cycloalkenyl, for example

or

), 0-3 R ^1d substituted C _6-10 aryl (the C _6-10 aryl such as phenyl) or 0-3 R ^1d substituted "having C _1-20 carbon atoms and 1-4 A heteroaryl group independently selected from N, NR ^1de1 , O or S(O) _0-2 heteroatoms" (the C _1-20 carbon atom is preferably a C _1-10 carbon atom, and the heteroaryl group is preferably Is a 5-12 membered heteroaryl group, for example

R ^5a为C _1-4烷基或C _1-4烷氧基；例如

可以为叔丁氧羰基)、氟； Each R ^1a and R ^{1b is} independently C _1-4 alkoxy, -C(=O)NR ^1de1 R ^1de2 , C _1-4 alkyl (e.g. methyl), C _1-4 haloalkyl (e.g. Trifluoromethyl), amino group, amino group protected by a protecting group (the protecting group can be

R ^5a is C _1-4 alkyl or C _1-4 alkoxy; for example

It can be tert-butoxycarbonyl), fluorine;

其中R ^5b为C _1-4烷基)； Each R ^{1c is} independently a C _1-4 alkyl or ester group (e.g.

Wherein R ^5b is C _1-4 alkyl);

或

)； Each R ^{1d is} independently halogen (e.g. fluorine, chlorine, bromine or iodine), R ^1a substituted or unsubstituted C _1-4 alkoxy (e.g. methoxy), R ^1a substituted or unsubstituted C _{1- 4} alkyl (the C _1-4 alkyl such as methyl, ethyl, n-propyl or isopropyl), -C(=O)NR ^1de1 R ^1de2 (the -C(=O)NR ^1de1 R ^1de2 such as -C(=O)NH ₂ ), -S(=O) ₂ R ^1de3 (the -S(=O) ₂ R ^{1de3 is} for example -S(=O) ₂ Me), NR ^1de1 R ^1de2 ( The NR ^1de1 R ^1de2 such as -NH ₂ ), R ^5c substituted or unsubstituted C _3-10 cycloalkyl (e.g. C _3-8 cycloalkyl), R ^5c substituted or unsubstituted "having C _1-20 Carbon atoms and 1-4 heterocyclic groups independently selected from NR ^1de1 , N, O or S(O) _0-2 heteroatoms" (the C _1-20 carbon atoms are preferably C _1-10 carbon atoms, so The heterocyclic group is preferably a 5-12 membered heterocyclic group, for example

or

);

Each R ^{5c is} independently halogen, C _1-4 alkyl, C _1-4 alkoxy, -C(=O)NR ^1de1 R ^1de2 or C _1-4 haloalkyl;

Each R ^{1de1 is} independently hydrogen or C _1-4 alkyl;

Each R ^{1de2 is} independently hydrogen or C _1-4 alkyl;

Each R ^1de3 is a C _1-4 alkyl group.

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

R ¹ is hydrogen, halogen, cyano, C _1-8 alkyl, C _1-8 haloalkyl, or any of the following structures:

among them,

Each j is independently 0, 1, 2 or 3; k is 0, 1, 2, 3 or 4; each V is independently C, N or O, and there are at most two N or at the same time in the same ring O; Ring A is a substituted or unsubstituted C _5-10 heteroaryl group containing 1-3 heteroatoms; each ring B is independently a substituted or unsubstituted C _5-10 containing 1-4 heteroatoms Heteroaryl, wherein the heteroatom is independently N, O, or S.

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

Each R ^{1a is} independently halogen (e.g. fluorine, chlorine, bromine or iodine) or hydroxyl.

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

Each R ^1a and R ^{1b is} independently -C(=O)OR ^1de4 (for example, -Boc);

R ^1de4 is a C _1-4 alkyl group (for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl).

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

或

)、0-3个R ^1d取代的C _6-10芳基(所述C _6-10芳基例如苯基)或0-3个R ^1d取代的“具有C _1-20个碳原子及1-4个独立选自N、NR ^1de1、O或 S(O) _0-2的杂原子的杂芳基”(所述C _1-20碳原子优选为C _1-10碳原子，所述杂芳基优选为5-12元杂芳基，更优选具有C _3-5碳原子及1-2个独立选自N和O的杂原子的杂芳基，例如呋喃基、吡啶基、吡唑基、异噻唑基或嘧啶基，又例如

R ¹ is R ^1b substituted or unsubstituted C _5-8 heterocycloalkenyl (the C _5-8 heterocycloalkenyl is preferably C having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur _5-8 heterocycloalkenyl, more preferably C _5-8 heterocycloalkenyl having one nitrogen, oxygen or sulfur heteroatom; for example

or

), 0-3 R ^1d substituted C _6-10 aryl (the C _6-10 aryl such as phenyl) or 0-3 R ^1d substituted "having C _1-20 carbon atoms and 1- 4 heteroaryl groups independently selected from N, NR ^1de1 , O or S(O) _0-2 heteroatoms" (the C _1-20 carbon atom is preferably a C _1-10 carbon atom, the heteroaryl group It is preferably a 5-12 membered heteroaryl group, more preferably a heteroaryl group having C _3-5 carbon atoms and 1-2 heteroatoms independently selected from N and O, such as furyl, pyridyl, pyrazolyl, iso Thiazolyl or pyrimidinyl, another example

Preferably, R ^1b is -C(=O)OR ^1de4 (for example, -Boc);

Preferably, the number of R ^1d is 0, 1, or 2;

)； Preferably, each R ^{1d is} independently halogen (such as fluorine, chlorine, bromine or iodine), R ^1a substituted or unsubstituted C _1-4 alkoxy (such as methoxy), R ^1a substituted or unsubstituted C _1-4 alkyl (for example methyl, ethyl, n-propyl or isopropyl), -C(=O)NR ^1de1 R ^1de2 (for example -C(=O)NH ₂ ), -S(=O ) ₂ R ^1de3 (for example -S(=O) ₂ Me), -NR ^1de1 R ^1de2 (for example -NH ₂ ) or R ^5c substituted or unsubstituted having C _1-20 carbon atoms and 1-4 independently selected (C _1-20 carbon atoms, preferably the C _1-10 carbon atoms ^NR 1de1, N, O, or S (O) heterocyclic group of _0-2 hetero atoms, the heterocyclic group preferably 5-12 membered heterocyclic ring Group, more preferably C _3-5 carbon atoms and 1-2 heterocyclic groups selected from N and O heteroatoms; for example

);

Preferably, each R ^{1a is} independently halogen or hydroxyl (e.g. fluorine, chlorine, bromine or iodine);

Preferably, R ^1de1 and R ^1de2 are hydrogen;

Preferably, R ^1de3 is methyl.

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

Each R ^{1d is} independently F, Cl, -OCH ₃ , -CH ₃ , -CF ₃ , -C(=O)NH ₂ , -S(=O) ₂ Me, -NH ₂ or

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

或

)； R ¹ is R ^1b substituted or unsubstituted C _5-8 heterocycloalkenyl (the C _5-8 heterocycloalkenyl is preferably C having 1 to 3 heteroatoms independently selected from nitrogen, oxygen and sulfur _5-8 heterocycloalkenyl, more preferably C _5-8 heterocycloalkenyl having one nitrogen, oxygen or sulfur heteroatom; for example

or

);

Preferably, R ^1b is -C(=O)OR ^1de4 (for example, -Boc).

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

R ¹ is a C _6-10 aryl group substituted with 0-3 R ^1d (the C _6-10 aryl group is for example phenyl),

Preferably, the number of R ^1d is 0, 1, or 2;

Preferably, each R ^{1d is} independently halogen (such as fluorine, chlorine, bromine or iodine), R ^1a substituted or unsubstituted C _1-4 alkoxy (such as methoxy), R ^1a substituted or unsubstituted C _1-4 alkyl (for example methyl, ethyl, n-propyl or isopropyl), -C(=O)NR ^1de1 R ^1de2 (for example -C(=O)NH ₂ ), -S(=O ) ₂ R ^1de3 (for example, -S(=O) ₂ Me) or NR ^1de1 R ^1de2 (for example, -NH ₂ );

Preferably, each R ^{1a is} independently halogen (such as fluorine, chlorine, bromine or iodine);

Preferably, R ^1de1 and R ^1de2 are hydrogen.

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

)； R ¹ is substituted with 0-3 R ^1d is "having C _1-20 carbon atoms and 1-4 heteroatoms independently selected from ^N, NR 1de1, O, or S (O) _0-2 heteroaryl group is an aryl group ( The C _1-20 carbon atoms are preferably C _1-10 carbon atoms, and the heteroaryl group is preferably a 5-12 membered heteroaryl group, more preferably having C _3-5 carbon atoms and 1-2 independently selected from N Heteroaryl groups with heteroatoms of O, such as furyl, pyridyl, pyrazolyl, isothiazolyl or pyrimidinyl, for example

);

Preferably, the number of R ^1d is 0, 1, or 2;

Preferably, each R ^{1d is} independently halogen (such as fluorine, chlorine, bromine or iodine), R ^1a substituted or unsubstituted C _1-4 alkoxy (such as methoxy), R ^1a substituted or unsubstituted C _1-4 alkyl (e.g. methyl, ethyl, n-propyl or isopropyl), NR ^1de1 R ^1de2 (e.g. -NH ₂ ) or R ^5c substituted or unsubstituted with C _1-20 carbon atoms and 1 -4 heterocyclic groups independently selected from NR ^1de1 , N, O or S(O) _0-2 heteroatoms (the C _1-20 carbon atoms are preferably C _1-10 carbon atoms, and the heterocyclic groups are preferably 5-12 membered heterocyclic group, more preferably C _3-5 carbon atoms and 1-2 heterocyclic groups selected from N and O heteroatoms; for example

Preferably, each R ^{1a is} independently halogen (such as fluorine, chlorine, bromine or iodine) and hydroxyl;

Preferably, R ^1de1 and R ^1de2 are hydrogen.

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

R ¹ is a single ring.

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

R ¹ is fluorine, bromine, iodine,

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

R ¹ is

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

In the compound represented by formula (I)

In some embodiments, the definitions of each letter and group in the compound of the above general formula are as follows, and the definitions of undefined letters and groups are as described above:

为

Group

for

In some embodiments, the compound represented by formula (I) is a compound represented by formula (Ia) below,

among them:

The definitions of R ¹ , R ² , R ³ and R ⁴ are as described above.

In some embodiments, the definitions of each letter and group in the compound represented by formula (Ia) are as follows, and the definitions of undefined letters and groups are as described above:

R ¹ is a C _6-10 aryl group substituted with 0-3 R ^1d (the C _6-10 aryl group is for example phenyl),

Preferably, the number of R ^1d is 0, 1, or 2;

Preferably, each R ^{1d is} independently halogen (such as fluorine, chlorine, bromine or iodine), R ^1a substituted or unsubstituted C _1-4 alkoxy (such as methoxy), R ^1a substituted or unsubstituted C _1-4 alkyl (for example methyl, ethyl, n-propyl or isopropyl), -C(=O)NR ^1de1 R ^1de2 (for example -C(=O)NH ₂ ) or -S(=O ) ₂ R ^1de3 (for example -S(=O) ₂ Me);

Preferably, each R ^{1a is} independently halogen (e.g. fluorine, chlorine, bromine or iodine).

In some embodiments, the definitions of each letter and group in the compound represented by formula (Ia) are as follows, and the definitions of undefined letters and groups are as described above:

)； R ¹ is substituted with 0-3 R ^1d is "having C _1-20 carbon atoms and 1-4 heteroatoms independently selected from ^N, NR 1de1, O, or S (O) _0-2 heteroaryl group is an aryl group ( The C _1-20 carbon atoms are preferably C _1-10 carbon atoms, and the heteroaryl group is preferably a 5-12 membered heteroaryl group, more preferably having C _3-5 carbon atoms and 1-2 independently selected from N Heteroaryl groups with heteroatoms of O, such as furyl, pyridyl, pyrazolyl, isothiazolyl or pyrimidinyl, for example

);

Preferably, the number of R ^1d is 0, 1, or 2;

)； Preferably, each R ^{1d is} independently halogen (such as fluorine, chlorine, bromine or iodine), R ^1a substituted or unsubstituted C _1-4 alkoxy (such as methoxy), R ^1a substituted or unsubstituted C _1-4 alkyl (such as methyl, ethyl, n-propyl or isopropyl), NR ^1de1 R ^1de2 (such as -NH ₂ ) or having C _1-20 carbon atoms and 1-4 independently selected from NR ^1de1 , N, O or S(O) _0-2 heteroatomic heterocyclic group (the C _1-20 carbon atom is preferably C _1-10 carbon atom, and the heterocyclic group is preferably 5-12 membered heterocyclic group , More preferably C _3-5 carbon atoms and 1-2 heterocyclic groups selected from N and O heteroatoms; for example

);

Preferably, each R ^{1a is} independently halogen (such as fluorine, chlorine, bromine or iodine) and hydroxyl;

Preferably, R ^1de1 and R ^1de2 are hydrogen.

In some embodiments, the definitions of each letter and group in the compound represented by formula (Ia) are as follows, and the definitions of undefined letters and groups are as described above:

R ¹ is

In some embodiments, the compound represented by formula (Ia) is selected from any of the following compounds:

Table 1

In some embodiments, the compound represented by formula (I) is a compound represented by formula (Ib) below,

Wherein R ¹ , R ² , R ³ and R ⁴ are as defined above.

In some embodiments, the definitions of each letter and group in the compound represented by formula (Ib) are as follows, and the definitions of undefined letters and groups are as described above:

R ¹ is 0-3 R ^1d substituted C _6-10 aryl (the C _6-10 aryl is for example phenyl) or 0-3 R ^1d substituted "having C _1-20 carbon atoms and 1 -4 heteroatoms independently selected from ^N, NR 1de1, O, or S (O) heteroaryl group of _0-2 hetero atoms "(the C _{1- 20} carbon atoms, preferably a C _1-10 carbon atoms, the heteroaryl The group is preferably a 5-12 membered heteroaryl group, more preferably a heteroaryl group having C _3-5 carbon atoms and 1-2 heteroatoms independently selected from N and O);

Preferably, the number of R ^1d is 0, 1, or 2;

Preferably, each R ^{1d is} independently halogen (for example, fluorine, chlorine, bromine or iodine), C _1-4 alkyl (for example, methyl, ethyl, n-propyl or isopropyl), -C(=O ) NR ^1de1 R ^1de2 (for example -C(=O)NH ₂ ) or -S(=O) ₂ R ^1de3 (for example, -S(=O) ₂ Me).

In some embodiments, the definitions of each letter and group in the compound represented by formula (Ib) are as follows, and the definitions of undefined letters and groups are as described above:

)； R ¹ is substituted with 0-3 R ^1d is "having C _1-20 carbon atoms and 1-4 heteroatoms independently selected from ^N, NR 1de1, O, or S (O) _0-2 heteroaryl group aryl" (The C _1-20 carbon atoms are preferably C _1-10 carbon atoms, and the heteroaryl group is preferably a 5-12 membered heteroaryl group, more preferably having C _3-5 carbon atoms and 1-2 independently selected from Heteroaryl groups of N and O heteroatoms, such as

);

Preferably, the number of R ^1d is 0, 1, or 2;

Preferably, each R ^{1d is} independently halogen (for example, fluorine, chlorine, bromine or iodine), C _1-4 alkyl (for example, methyl, ethyl, n-propyl or isopropyl), -C(=O ) NR ^1de1 R ^1de2 (for example -C(=O)NH ₂ ) or -S(=O) ₂ R ^1de3 (for example, -S(=O) ₂ Me).

In some embodiments, the definitions of each letter and group in the compound represented by formula (Ib) are as follows, and the definitions of undefined letters and groups are as described above:

R ¹ is

In some embodiments, the compound represented by formula (Ib) is selected from any of the following compounds:

Table 2

The present invention further provides a compound represented by formula (I), compounds isotopically-labeled pharmaceutically acceptable salt thereof, a hydrate thereof, a prodrug, a stereoisomer or a solvate thereof, wherein the isotope is selected from ² H , ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ Cl and ¹²⁵ I. The atoms that can be isotopically labeled in the compound represented by formula (I) include, but are not limited to, hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, etc., which can be isotopes ² H, ³ H, ¹¹ C, respectively. , ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ Cl and ¹²⁵ I etc. instead.

The present invention also provides a preparation method of the compound represented by formula (I), which comprises the following steps:

Wherein, W represents halogen, preferably Br; X, n, R ¹ , R ² , R ³ and R ⁴ are as defined above.

In some embodiments, the method for preparing the compound represented by formula (I) includes the following steps: the halogenated intermediate A and the compound R ¹ -W ¹ undergo the Suzuki reaction shown below;

)。 Wherein, W represents halogen (such as fluorine, chlorine, bromine or iodine), preferably Br; X, n, R ¹ , R ² , R ³ and R ⁴ are as defined above, and W ¹ is boric acid, boric acid ester or boron Acid salt (e.g. -B(OH) ₂ or

).

The method for preparing the compound represented by formula (I) is preferably carried out under gas protection, in a solvent, and in the presence of a base. Among them, the solvent is preferably halogenated hydrocarbon solvents (such as one or more of dichloromethane, chloroform and 1,2-dichloroethane), ether solvents (such as dioxane and/or tetrahydrofuran). ), one or more of amide solvents (such as DMF), alkanone solvents (such as NMP), nitrile solvents (such as acetonitrile), and sulfoxide solvents (such as DMSO).

The gas in the gas protection is preferably nitrogen.

In the selected examples, the preparation method of compound I-A includes the following steps:

The halogenated intermediate A and the corresponding boronic acid, boronic acid ester or boronic acid salt are coupled to produce compound I-A.

Where R ^{1A is} selected from

W, X, n, j, k, R ^1b , R ^1c , R ^1d , R ² , R ³ , R ⁴ , ring A and ring B are as defined above.

The present invention also provides a preparation method of compound A, which comprises the following steps:

The chlorinated intermediate B is substituted with the intermediate amine C under basic conditions to obtain the halogenated A.

The reaction equation is as follows:

Wherein, W, X, n, R ² , R ³ and R ⁴ are as defined above.

The present invention also provides a method for preparing compound C1 and its salt, and the reaction process is as follows:

Compound C1-1 is reacted with 1,2 dihaloethane (such as 1,2 dibromoethane) to obtain compound C1-2, and C1-2 undergoes elimination reaction (such as potassium tert-butoxide treatment) to obtain compound C1-3 , C1-3 is condensed with p-toluenesulfonyl hydrazide to obtain sulfonyl hydrazone C1-4, and C1-4 is reacted with furo three-membered ring to obtain compound C1-5. The bromine atom of C1-5 is catalyzed by cyano group substitution to obtain compound For C1-6, the cyano group is reduced to an amine group and protected with in-situ Boc anhydride to obtain C1-7, and C1-7 is removed to obtain C1.

Wherein, Z represents halogen (e.g. chlorine, bromine or iodine).

The solvent involved in the present invention is selected from: dichloromethane, chloroform, 1,2-dichloroethane, dioxane, DMF, acetonitrile, DMSO, NMP, THF or a combination thereof.

The bases involved in the present invention include organic bases and inorganic bases.

The organic base involved in the present invention is selected from TEA, DIPEA or a combination thereof.

The inorganic base involved in the present invention is selected from sodium hydride, potassium carbonate, sodium carbonate, cesium carbonate, potassium tert-butoxide, sodium tert-butoxide, LiHMDS, LDA, butyl lithium or a combination thereof.

The isotopically labeled compound of the compound represented by the formula (I) of the present invention can be prepared by a synthetic method similar to that of the unlabeled compound, except that the unlabeled starting materials and/or reagents are replaced with isotopically labeled ones. Starting materials and/or reagents.

The present invention also provides a pharmaceutical composition comprising the compound represented by formula (I) as described above, its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer or its solvent Compound or the aforementioned isotope-labeled compound and pharmaceutically acceptable excipients. The pharmaceutically acceptable auxiliary materials may be one or more of diluents, absorbents, wetting agents, binders, disintegrants and lubricants.

The present invention also provides the compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer or its solvate or the aforementioned isotope-labeled compound, or Application of the above-mentioned pharmaceutical composition in preparing medicine.

The drug is preferably an EED inhibitor or an EZH2 inhibitor or a PRC2 inhibitor. The drugs can be used to treat cancer.

The present invention also provides the compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer or its solvate or the aforementioned isotope-labeled compound, or Application of the above-mentioned pharmaceutical composition in the preparation of cancer drugs related to the mechanism of action of EED protein and/or PRC2 protein complex.

Preferably, the cancer in the present invention includes, but is not limited to, lymphoma such as diffuse large B-cell lymphoma, follicular lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, liver cancer , Prostate cancer, breast cancer, brain tumors including neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, Lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, kidney cancer, rectal cancer, thyroid cancer, parathyroid tumor, uterine tumor and soft tissue sarcoma, etc.

The present invention also provides the compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer or its solvate or the aforementioned isotope-labeled compound, or Application of the above-mentioned pharmaceutical composition in preparing EED inhibitor, EZH2 inhibitor or PRC2 inhibitor.

Preferably, the compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer or its solvate or the aforementioned isotope-labeled compound is used in combination with other drugs; More preferably, the other drugs are selected from anticancer drugs, tumor immune drugs, anti-allergic drugs, antiemetics, analgesics, and cytoprotective drugs.

The present invention also provides a pharmaceutical preparation comprising the compound represented by formula (I) as described above, its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer or its solvate Or the aforementioned isotope-labeled compound, or the pharmaceutical composition as described above, can be taken in a suitable manner, such as tablets, capsules (such as sustained release or timed release capsules), pills, powders, granules (such as small particles), elixirs Suspensions, syrups, emulsions, solutions, etc., in the form of suspensions, syrups, emulsions, solutions, etc., can be used for oral administration, sublingual administration, including subcutaneous injection, suspensions (such as nanosuspension, microsuspension) and spray-dried dispersions. Intravenous injection, intramuscular injection, intrasternal injection, injection and other forms of injection, nasal administration (such as nasal membrane inhalation), topical surface (such as creams and ointments), rectal administration (such as suppositories), etc. The compound disclosed in the present invention can be taken alone or together with an appropriate drug carrier.

The present invention also provides that the aforementioned pharmaceutical preparations can be formulated into appropriate pharmaceutical dosages to facilitate and control the dosage of the pharmaceuticals. The dosage regimen of the compound disclosed in the present invention varies according to specific factors, such as the pharmacodynamics and the way of taking, the subject, gender, age, health status and the weight of the subject, disease characteristics, other simultaneous medication status, and medication Frequency, liver and kidney function and desired effect, etc. The compound disclosed in the present invention can be taken in a single dose per day, or the total dose can be divided into multiple doses (for example, two to four times a day).

The present invention also provides a method of treating cancer, which comprises administering a therapeutically effective amount of the compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, Its prodrug, its stereoisomer or its solvate or the aforementioned isotope-labeled compound, or the pharmaceutical composition as described above. The cancer includes, but is not limited to, lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, liver cancer, prostate cancer, breast cancer such as diffuse large B-cell lymphoma, follicular lymphoma, etc. Cancer and brain tumors include neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal cancer , Ovarian cancer, pancreatic cancer, kidney cancer, rectal cancer, thyroid cancer, parathyroid tumor, uterine tumor and soft tissue sarcoma, etc. In the present invention, the cancer is preferably cancer caused by abnormal function of PRC2.

The present invention also provides the compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer or its solvate or the aforementioned isotope-labeled compound used in combination with other drugs, The other drugs are selected from: anticancer drugs, tumor immune drugs, anti-allergic drugs, antiemetics, analgesics, cytoprotective drugs, etc., and the combined use has better effects.

It should be understood that, within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as the embodiments) can be combined with each other to form a new or preferred technical solution.

Term Description

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention belongs.

As used herein, when used in reference to a specifically recited value, the term "about" means that the value can vary from the recited value by no more than 1%. For example, as used herein, the expression "about 100" includes all values between 99 and 101 (eg, 99.1, 99.2, 99.3, 99.4, etc.).

As used herein, the term "containing" or "including (including)" can be open, semi-closed, and closed. In other words, the term also includes "essentially composed of" or "consisting of".

Group definition

Definitions of standard chemical terms can be found in references (including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4TH ED." Vols.A (2000) and B (2001), Plenum Press, New York). Unless otherwise specified, conventional methods within the technical scope of the art, such as mass spectrometry, NMR, IR, and UV/VIS spectroscopy and pharmacological methods, are used. Unless specific definitions are provided, the terms used in the description of analytical chemistry, synthetic organic chemistry, and pharmaceuticals and medicinal chemistry herein are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and treatment of patients. For example, the manufacturer's instructions for use of the kit can be used, or the reaction and purification can be performed in a manner known in the art or the instructions of the present invention. Generally, the above-mentioned techniques and methods can be implemented according to the descriptions in a number of summary and more specific documents cited and discussed in this specification according to conventional methods well known in the art. In this specification, groups and their substituents can be selected by those skilled in the art to provide stable structural parts and compounds.

When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, -CH ₂ O- is equivalent to -OCH ₂ -.

The chapter headings used in this article are only for the purpose of organizing the article, and should not be construed as a limitation on the subject. All documents or document parts cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals and theses, are incorporated herein by reference in their entirety.

Certain chemical groups defined herein are preceded by simplified symbols to indicate the total number of carbon atoms present in the group. For example, C _1-6 alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms (such as 1, 2, 3, 4, 5, 6 carbon atoms). The total number of carbon atoms in the simplified notation does not include carbons that may be present in the substituents of the group.

In addition to the foregoing, when used in the specification and claims of this application, unless otherwise specified, the following terms have the following meanings.

In this application, the term "halogen" refers to fluorine, chlorine, bromine or iodine.

"Hydroxy" refers to the -OH group.

中的R为羰基时，

"Carbonyl" refers to the -C(=O)- group. when

When R is a carbonyl group,

"Cyano" refers to -CN.

"Amino" refers to -NH ₂ .

R可以为烷基、烯基、芳基、杂芳基、环烷基、环烯基、芳基烷基或杂芳基烷基等。 "Ester group" means

R can be alkyl, alkenyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, arylalkyl or heteroarylalkyl, and the like.

每个R可以独立地为氢、烷基、烯基、芳基、杂芳基、环烷基、环烯基、芳基烷基或杂芳基烷基等。 "Amide" means

Each R can independently be hydrogen, alkyl, alkenyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, arylalkyl, or heteroarylalkyl, and the like.

"Substituted amino group" refers to an amino group substituted with one or two alkyl groups, alkylcarbonyl groups, aralkyl groups, heteroaralkyl groups as defined below, for example, monoalkylamino, dialkylamino, alkyl Amido, aralkylamino, heteroaralkylamino.

"Carboxy" refers to -COOH.

In this application, as a group or a part of other groups (for example, used in halogen-substituted alkyl groups and other groups), the term "alkyl" refers to a fully saturated linear or branched hydrocarbon chain group, Consists of only carbon atoms and hydrogen atoms, has, for example, 1 to 12 (preferably 1 to 8, more preferably 1 to 6, more preferably 1 to 4) carbon atoms, and is connected to the rest of the molecule through a single bond Partial connections, such as but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2 , 2-Dimethylpropyl, n-hexyl, heptyl, 2-methylhexyl, 3-methylhexyl, octyl, nonyl and decyl, etc.

In this application, as a group or part of other groups, the term "haloalkyl" means that one or more hydrogen atoms in an alkyl group (as defined in the present invention) are halogenated (as defined in the present invention). ), the number of halogens can be one or more; when the number of halogens is more than one, the halogens are the same or different. For example, fluoroalkyl means that the alkyl group is substituted with one or more fluorines. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl and difluoromethyl.

In this application, as a group or a part of other groups, the term "alkenyl" means consisting only of carbon atoms and hydrogen atoms, containing at least one double bond, having, for example, 2 to 14 (preferably 2 to 10 One, more preferably 2 to 6, more preferably 2 to 4) a straight or branched hydrocarbon chain group connected to the rest of the molecule through a single bond, such as but not limited to vinyl, propylene Group, allyl group, but-1-enyl group, but-2-enyl group, pent-1-enyl group, pent-1-enyl group, etc.

In this application, as a group or a part of other groups, the term "cyclic hydrocarbon group" means a stable non-aromatic monocyclic or polycyclic hydrocarbon group composed of only carbon atoms and hydrogen atoms, which may include fused rings System, bridged ring system or spiro ring system, having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, more preferably 3 to 8 carbon atoms, and it is saturated or unsaturated and can be passed through any suitable The carbon atom is connected to the rest of the molecule by a single bond. Unless specifically indicated otherwise in this specification, the carbon atoms in the cyclic hydrocarbon group may be optionally oxidized. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, 1H-indene Group, indanyl, 1,2,3,4-tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, 8,9-dihydro-7H-benzo Cyclohepten-6-yl, 6,7,8,9-tetrahydro-5H-benzocycloheptenyl, 5,6,7,8,9,10-hexahydro-benzocyclooctenyl, Fluorenyl, bicyclo[2.2.1]heptyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, bicyclo[2.2.2]octyl Group, bicyclo[3.1.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octenyl, bicyclo[3.2.1]octenyl, adamantyl, octahydro -4,7-methylene-1H-indenyl and octahydro-2,5-methylene- and cyclopentadienyl, etc.

In this application, as a group or part of another group, the term "cycloalkyl" means a saturated cyclic hydrocarbon group.

In this application, as a group or a part of other groups, the term "cycloalkenyl" means a cyclic hydrocarbon group having at least one double bond (such as a carbon-carbon double bond). The cycloalkenyl group can be connected to the rest of the molecule through the atom of the double bond.

In this application, as a group or part of another group, the term "heterocyclic group" means a group consisting of 2 to 14 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, phosphorus, oxygen and sulfur Stable 3- to 20-membered non-aromatic cyclic group. Unless otherwise specified in the specification, the heterocyclic group may be a monocyclic, bicyclic, tricyclic or more ring system, which may include a fused ring system, a bridged ring system or a spiro ring system; in the heterocyclic group The nitrogen, carbon, or sulfur atoms of may be optionally oxidized; the nitrogen atom may be optionally quaternized; and the heterocyclic group may be partially or fully saturated. The heterocyclic group can be connected to the rest of the molecule via a carbon atom or a heteroatom and through a single bond. In heterocyclic groups containing fused rings, one or more rings may be aryl or heteroaryl as defined below, provided that the point of attachment to the rest of the molecule is a non-aromatic ring atom. For the purpose of the present invention, the heterocyclic group is preferably a stable 4- to 11-membered non-aromatic monocyclic, bicyclic, bridged ring or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur The group is more preferably a stable 4- to 8-membered non-aromatic monocyclic, bicyclic, bridged ring or spirocyclic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heterocyclic groups include, but are not limited to: pyrrolidinyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, thiomorpholinyl, 2,7-diaza-spiro[3.5]non Alkyl-7-yl, 2-oxa-6-aza-spiro[3.3]heptane-6-yl, 2,5-diaza-bicyclo[2.2.1]heptan-2-yl, aza Cyclobutanyl, pyranyl, tetrahydropyranyl, thiopyranyl, tetrahydrofuranyl, oxazinyl, dioxopentyl, tetrahydroisoquinolinyl, decahydroisoquinolinyl, imidazolinyl, Imidazolidinyl, quinazinyl, thiazolidinyl, isothiazolidinyl, isoxazolidinyl, indoline, octahydroindolyl, octahydroisoindolyl, pyrrolidinyl, pyrazolidinyl , Phthalimide, etc.

In this application, as a group or part of another group, the term "heteroalkyl" means a C _3-8 heteroalkyl group having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

In the present application, as a group or a part of other groups, the term "aryl" means a conjugated hydrocarbon ring system group having 6 to 18 carbon atoms (preferably having 6 to 10 carbon atoms). For the purpose of the present invention, the aryl group can be a monocyclic, bicyclic, tricyclic or more cyclic ring system, and can also be fused with a cycloalkyl or heterocyclic group as defined above, provided that the aryl group passes through The atoms on the aromatic ring are connected to the rest of the molecule through a single bond. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthryl, fluorenyl, 2,3-dihydro-1H-isoindolyl, 2-benzoxazolinone, 2H-1, 4-benzoxazine-3(4H)-one-7-yl and the like.

In this application, the term "arylalkyl" refers to the above-defined alkyl group substituted by the above-defined aryl group.

In this application, as a group or part of other groups, the term "heteroaryl" means having 1 to 15 carbon atoms (preferably having 1 to 10 carbon atoms) and 1 to 6 nitrogen atoms in the ring. 5 to 16 membered conjugated ring system group of heteroatoms of, oxygen and sulfur. Unless otherwise specified in this specification, heteroaryl groups can be monocyclic, bicyclic, tricyclic or more cyclic ring systems, and can also be fused with cycloalkyl or heterocyclic groups as defined above, provided that the hetero The aryl group is connected to the rest of the molecule via a single bond through an atom on the aromatic ring. The nitrogen, carbon or sulfur atoms in the heteroaryl group can be optionally oxidized; the nitrogen atom can be optionally quaternized. For the purpose of the present invention, the heteroaryl group is preferably a stable 5- to 12-membered aromatic group containing 1 to 5 heteroatoms selected from nitrogen, oxygen and sulfur, and more preferably contains 1 to 4 selected heteroatoms. A stable 5- to 10-membered aromatic group from heteroatoms of nitrogen, oxygen, and sulfur or a 5- to 6-membered aromatic group containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl groups include, but are not limited to, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, oxadiazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, Benzimidazolyl, benzopyrazolyl, indolyl, furyl, pyrrolyl, triazolyl, tetrazolyl, triazinyl, indazinyl, isoindolyl, indazolyl, isoindazolyl , Purinyl, Quinolinyl, Isoquinolinyl, Diazonaphthyl, Naphthyridinyl, Quinoxolinyl, Pteridinyl, Carbazolyl, Carboline, Phenanthridinyl, Phenanthrolinyl, Acridine Group, phenazinyl, isothiazolyl, benzothiazolyl, benzothienyl, oxtriazolyl, cinnolinyl, quinazolinyl, phenylthio, indolizinyl, phenanthroline, Isoxazolyl, phenoxazinyl, phenothiazinyl, 4,5,6,7-tetrahydrobenzo[b]thienyl, naphthopyridyl, [1,2,4]triazolo[4 ,3-b]pyridazine, [1,2,4]triazolo[4,3-a]pyrazine, [1,2,4]triazolo[4,3-c]pyrimidine, [1, 2,4]Triazolo[4,3-a]pyridine, imidazo[1,2-a]pyridine, imidazo[1,2-b]pyridazine, imidazo[1,2-a]pyrazine Wait.

In this application, the term "heteroarylalkyl" refers to the above-defined alkyl group substituted by the above-defined heteroaryl group.

In this application, "optionally" means that the event or condition described later may or may not occur, and the description includes both occurrence and non-occurrence of the event or condition. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted aryl groups and unsubstituted aryl groups. The "optionally" substituents described in the claims and specification of the present invention are selected from alkyl, alkenyl, alkynyl, halogen, halogenated alkyl, halogenated alkenyl, halogenated alkynyl, cyano, nitro , Optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, optionally substituted heterocyclic alkyl.

In the present invention, the term "substituted" or "substituent" means that one or more hydrogen atoms are replaced by a designated group. When the number of substituents is not specified, the substituents can be one or more; when the substitution position is not specified, the substitution can be at any position, but only the formation of a stable or chemically feasible chemical can be Allowed.

中，当n为2时，表示苯环被2个R取代，并且每个R都有独立的选项，即2个R可以相同，也可以不同。此外，取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。 When any variable (such as R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group can optionally be substituted with up to two Rs, and R has independent options in each case. For example, in

In, when n is 2, it means that the benzene ring is replaced by two Rs, and each R has independent options, that is, the two Rs can be the same or different. In addition, combinations of substituents and/or variants thereof are only permitted if such combinations result in stable compounds.

The terms "part", "structural part", "chemical part", "group", and "chemical group" as used herein refer to specific fragments or functional groups in a molecule. The chemical moiety is generally considered to be a chemical entity embedded in or attached to a molecule.

When the compound of the present invention contains an olefinic double bond, unless otherwise specified, the compound of the present invention is intended to include E- and Z-geometric isomers.

"Tautomer" refers to an isomer formed by transferring a proton from one atom of a molecule to another atom of the same molecule. All tautomeric forms of the compounds of the present invention will also be included in the scope of the present invention.

The compound of the present invention or a pharmaceutically acceptable salt thereof may contain one or more chiral carbon atoms, and therefore may produce enantiomers, diastereomers and other stereoisomeric forms. Each chiral carbon atom can be defined as (R)- or (S)- based on stereochemistry. The present invention is intended to include all possible isomers, as well as their racemates and optically pure forms. The preparation of the compounds of the present invention can select racemates, diastereomers or enantiomers as raw materials or intermediates. Optically active isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

Conventional techniques for preparing/separating individual isomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives using, for example, chiral high performance liquid chromatography) ), for example, see Gerald Gübitz and Martin G. Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol. 243, 2004; AMStalcup, Chiral Separations, Annu. Rev. Anal. Chem. 3 :341-63,2010; Fumis et al.(eds.),VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.TH ED.,Longman Scientific and Technical Ltd.,Essex,1991,809-816;Heller,Acc.Chem .Res.1990,23,128.

In this application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"Pharmaceutically acceptable acid addition salt" refers to a salt formed with an inorganic acid or an organic acid that can retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc.; organic acid salts include, but are not limited to, formate, acetate, and 2,2-dichloroacetate , Trifluoroacetate, propionate, caproate, caprylate, caprate, undecylenate, glycolate, gluconate, lactate, sebacate, hexanoate Acid salt, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate , Cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate , Alginate, ascorbate, salicylate, 4-aminosalicylate, naphthalene disulfonate, etc. These salts can be prepared by methods known in the art.

"Pharmaceutically acceptable base addition salt" refers to a salt formed with an inorganic base or an organic base that can maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Preferred inorganic salts are ammonium, sodium, potassium, calcium and magnesium salts. Salts derived from organic bases include but are not limited to the following salts: primary amines, secondary amines and tertiary amines, substituted amines, including natural substituted amines, cyclic amines and basic ion exchange resins , Such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, bicyclic Hexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperazine Pyridine, N-ethylpiperidine, polyamine resin, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. These salts can be prepared by methods known in the art.

In this application, "pharmaceutical composition" refers to a preparation of the compound of the present invention and a medium generally accepted in the art for delivering the biologically active compound to a mammal (such as a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote the administration of the organism, facilitate the absorption of the active ingredients and then exert the biological activity.

The term "pharmaceutically acceptable" as used herein refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compound of the present invention, and is relatively non-toxic, that is, the substance can be administered to an individual without causing undesirable biological activity. Reacts or interacts in an undesirable manner with any components included in the composition.

In this application, "pharmaceutically acceptable excipients" include, but are not limited to, any adjuvants, carriers, excipients, glidants, enhancers that are approved by relevant government authorities as acceptable for human or livestock use. Sweeteners, diluents, preservatives, dyes/colorants, flavors, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents or emulsifiers.

The "tumor" of the present invention includes but is not limited to brain tumors including neuroblastoma, glioma, glioblastoma and astrocytoma, sarcoma, melanoma, articular chondroma, cholangiocarcinoma, leukemia, gastrointestinal Stromal tumor, diffuse large B-cell lymphoma, follicular lymphoma and other lymphomas, histiocytic lymphoma, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, lung squamous cell carcinoma, lung adenocarcinoma, breast cancer, prostate Cancer, liver cancer, skin cancer, epithelial cell cancer, cervical cancer, ovarian cancer, bowel cancer, nasopharyngeal cancer, brain cancer, bone cancer, esophageal cancer, melanoma, kidney cancer, oral cancer, multiple myeloma, mesothelioma , Malignant rhabdoid tumor, endometrial cancer, head and neck cancer, thyroid cancer, parathyroid tumor, uterine tumor and soft tissue sarcoma and other diseases.

The terms "preventive", "preventing" and "preventing" as used herein include reducing the likelihood of a disease or condition from occurring or worsening.

As used herein, the term "treatment" and other similar synonyms include the following meanings:

(i) Preventing the occurrence of a disease or condition in a mammal, especially when such a mammal is susceptible to the disease or condition, but has not been diagnosed as having the disease or condition;

(ii) To suppress the disease or disease, that is, to curb its development;

(iii) Alleviate the disease or condition, that is, make the state of the disease or condition subside; or

(iv) Reducing the symptoms caused by the disease or condition.

The term "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" as used herein refers to at least one agent or compound that is sufficient to relieve one or more symptoms of the disease or condition being treated after administration The amount. The result can be a reduction and/or alleviation of signs, symptoms or causes, or any other desired changes in the biological system. For example, the "effective amount" for treatment is the amount of the composition containing the compound disclosed herein that is required to provide significant disease relief clinically. Techniques such as dose escalation tests can be used to determine the effective amount suitable for any individual case.

As used herein, the terms "administration", "administration", "administration" and the like refer to methods capable of delivering a compound or composition to a desired site for biological action. These methods include, but are not limited to, oral routes, transduodenal routes, parenteral injections (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. Those skilled in the art are familiar with the application techniques that can be used for the compounds and methods described herein, for example in Goodman and Gilman, The Pharmaceutical Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical (current edition), Mack Publishing Co., Those discussed in Easton, Pa. In a preferred embodiment, the compounds and compositions discussed herein are administered orally.

The terms "drug combination", "drug combination", "combination drug", "administration of other treatments", "administration of other therapeutic agents" and the like as used herein refer to drug treatments obtained by mixing or combining more than one active ingredient. It includes fixed and non-fixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration of at least one compound described herein and at least one synergistic agent to a patient in the form of a single entity or a single dosage form. The term "non-fixed combination" refers to the simultaneous administration, co-administration or sequential administration of at least one compound described herein and at least one synergistic agent to a patient in the form of separate entities. These also apply to cocktail therapy, such as the administration of three or more active ingredients.

Those skilled in the art should also understand that in the methods described below, the functional group of the intermediate compound may need to be protected by an appropriate protecting group. Such functional groups include hydroxyl, amino, mercapto and carboxylic acid. Suitable hydroxy protecting groups include trialkylsilyl or diarylalkylsilyl groups (e.g., tert-butyldimethylsilyl, tert-butyldiphenylsilyl, or trimethylsilyl) , Tetrahydropyranyl, benzyl, etc. Suitable protecting groups for amino, amidino and guanidino include tert-butoxycarbonyl, benzyloxycarbonyl and the like. Suitable sulfhydryl protecting groups include -C(O)-R" (wherein R" is alkyl, aryl or aralkyl), p-methoxybenzyl, trityl and the like. Suitable carboxy protecting groups include alkyl, aryl or aralkyl esters.

Protecting groups can be introduced and removed according to standard techniques known to those skilled in the art and as described herein. The use of protective groups is detailed in Greene, T.W. and P.G.M. Wuts, Protective Groups in Organi Synthesis, (1999), 4th Ed., Wiley. The protecting group can also be a polymer resin.

detailed description

The present invention will be further explained by way of examples below, but the present invention is not limited to the scope of the described examples. In the following examples, the experimental methods without specific conditions are selected according to conventional methods and conditions, or according to the product specification.

The starting materials used in the following examples can be purchased from chemical vendors such as Aldrich, TCI, Alfa Aesar, Beide, Anaiji, etc., or can be synthesized by known methods.

In the following examples, the ice bath refers to -5°C to 0°C, the room temperature refers to 10°C to 30°C, and the reflux temperature generally refers to the solvent reflux temperature under normal pressure. An overnight reaction means that the time is 8-15 hours. In the following examples, if the specific operating temperature is not limited, it is all carried out at room temperature.

In the following examples, the separation and purification of the intermediate and the final product is by normal phase or reverse phase chromatography column separation or other suitable methods. The normal phase flash chromatography column uses ethyl acetate and n-hexane or methanol and dichloromethane as the mobile phase. Reversed-phase preparative high pressure liquid chromatography (HPLC) uses a C18 column and uses UV214nm and 254nm to detect, and its mobile phase is A (water and 0.1% formic acid), B (acetonitrile) or mobile phase A (water and 0.1% hydrogen carbonate) Ammonium), B (acetonitrile).

In each embodiment:

LCMS instrument: Pump Agilent 1260 UV detector: Agilent 1260 DAD

Mass Spectrometer API 3000

Chromatography column: Waters sunfire C18, 4.6×50mm, 5um

Mobile phase: AH ₂ O (0.1% HCOOH); B-acetonitrile

NMR instrument: Bruker Ascend 400M ( ¹ H NMR: 400 MHz; ¹³ C NMR: 100 MHz).

Example 1: 8-Bromo-5-chloro-[1,2,4]triazolo[4,3-c]pyrimidine (B1)

Step 1: Synthesis of 5-bromo-2-chloro-4-hydrazinopyrimidine (B1-b):

Add 5-bromo-2-chloro-4-chloropyrimidine (B1-a) (2g, 8.78mmol) and ethanol (20mL) into a 50mL single-neck flask, and slowly add hydrazine hydrate (1.72g, 53.65mmol) in an ice bath. The suspension was stirred at 60°C for 3 hours. After the reaction, it was cooled to room temperature and a light yellow solid precipitated. The solid was collected by filtration. The filter cake was washed with ethanol (5 mL) and dried to obtain 5-bromo-2-chloro-4-hydrazinopyrimidine (B1-b) (1.8g, yield 92%).

¹ H NMR (DMSO-d6, 400MHz) δ 8.06 (s, 1H), 7.85 (s, 1H), 4.34 (s, 2H) ppm.

Step 2: Synthesis of 5-bromo-8-chloro-[1,2,4]triazolo[4,3-a]pyrazine (B1):

B1-b (1.2 g, 5.37 mmol), trimethyl orthoformate (12 mL), and trifluoroacetic acid (1 drop) were added to a 50 mL single-necked flask, and the temperature was raised to 100° C. to react for 10 h. After the reaction, the temperature was lowered to room temperature, the trimethyl orthoformate was removed by rotary evaporation, and the concentrate was purified with a silica gel column (PE:EA=20:1) to obtain the yellow solid product 8-bromo-5-chloro-[1,2,4 ] Triazolo[4,3-c]pyrimidine B1 (960 mg, yield 77%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.03 (s, 1H), 8.05 (s, 1H) ppm; LCMS: m/z 232.9 [M+H] ⁺ .

Example 2: Intermediate 5-chloro-8-iodopyrido[4,3-d]pyrimidine (B2):

Step 1: pyrido[4,3-d]pyrimidine-5(6H)-one (B2-b)

Add 200mL of ethanol and sodium metal (2.48g, 107.92mmol) in a dry 500mL single-necked flask. When the sodium metal is completely reacted, add 1,3,5 triazine (B2-a) (35g, 431.68mmol) in sequence And ethyl acetoacetate (51.68 g, 431.68 mmol). The reaction temperature was raised to 85°C and the reaction was stirred for 3 hours. After the reaction, the reaction solution was spin-dried and poured into 100 mL of ice water, and the pH of the reaction solution was adjusted to 3-4 with concentrated hydrochloric acid solution. The precipitated solid was filtered and washed with a small amount of ice acetone to obtain a yellow solid product pyrido[4,3-d]pyrimidin-5(6H)-one (B2-b) (7.0 g, yield: 11.0%).

¹ H NMR(400MHz,DMSO-d ₆ )δ11.90(s,1H), 9.42(s,1H), 9.33(s,1H), 7.72(d,J=7.3Hz,1H), 6.57(d, J = 7.3 Hz, 1H) ppm; LCMS: m/z148.1[M+H] ⁺ .

Step 2: 8-Iodopyrido[4,3-d]pyrimidine-5(6H)-one (B2-c)

In a dry 250mL single-neck flask, add 150mL of 0.4M sodium hydroxide aqueous solution, iodine (12.56g, 49.48mmol) and pyrido[4,3-d]pyrimidin-5(6H)-one (B2-b) in sequence (5.60g, 38.06mmol). The reaction temperature was raised to 80°C and the reaction was stirred for 18 hours. After the reaction was completed, the precipitated solid was filtered and washed with a small amount of water to obtain a yellow solid product 8-iodopyrido[4,3-d]pyrimidin-5(6H)-one (B2-c) (8.0g, 77% yield) rate).

¹ H NMR(400MHz,DMSO-d ₆ )δ12.17(s,1H),9.43(s,1H),9.33(s,1H), 8.14(s,1H)ppm; LCMS: m/z274.1[ M+H] ⁺ .

Step 3: 5-chloro-8-iodopyrido[4,3-d]pyrimidine (B2)

In a dry 250 mL single-neck flask, 100 mL of phosphorus oxychloride and 8-iodopyrido[4,3-d]pyrimidin-5(6H)-one (B2-c) (8.0 g, 29.30 mmol) were sequentially added. The reaction temperature was raised to 110°C and the reaction was stirred for 24 hours. After the completion of the reaction, the reaction solution was spin-dried and poured into 100 mL ice water, and extracted with ethyl acetate (50 mL × 3). The organic phase was washed sequentially with water (20 mL×1) and saturated brine (20 mL×1). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Purified by column chromatography (ethyl acetate: petroleum ether = 1:10) to obtain a yellow solid 5-chloro-8-iodopyrido[4,3-d]pyrimidine (B2) (4.5g, 52.7% yield) .

¹ H NMR(400MHz,CDCl ₃ )δ9.75(s,1H),9.64(s,1H),9.12(s,1H)ppm; LCMS: m/z292.0[M+H] ⁺ .

Example 3: (5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methylamine (C1)

Step 1: Intermediate 2-bromo-6-(2-bromoethoxy)-3-fluorobenzaldehyde (C1-b):

Add 2-bromo-3-fluoro-6-hydroxybenzaldehyde (C1-a) (19g, 86.7mmol) into a 250mL single-neck flask, add dry DMF (90mL) and stir to dissolve, add potassium carbonate (24g, 173.5mmol) ), 1,2-dibromoethane (24g, 130.1mmol), heated to 64°C, and stirred for 18 hours. After the reaction, it was cooled to room temperature, diluted with ethyl acetate (400 mL), stirred for 15 minutes, filtered to remove insoluble salts, the filter cake was washed with ethyl acetate (100 mL) once, and the filtrate was washed with saturated sodium chloride (100 mL) twice The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was quickly purified by a short silica gel column (petroleum ether: ethyl acetate = 50:1) to obtain the intermediate 2-bromo-6-(2-bromo Ethoxy)-3-fluorobenzaldehyde (C1-b) (25 g, 88% yield) as a yellow solid.

¹ H NMR (400MHz, CDCl ₃ ) δ 10.52-10.27 (m, 1H), 7.40-7.09 (m, 1H), 7.02-6.78 (m, 1H), 4.53-4.05 (m, 2H), 3.68 (t ,J=6.0Hz,2H)ppm.

Step 2: Intermediate 2-bromo-3-fluoro-6-(vinyloxy)benzaldehyde (C1-c):

Add 2-bromo-6-(2-bromoethoxy)-3-fluorobenzaldehyde (C1-b) (10g, 30.7mmol) in a 1L single-neck flask, add anhydrous tetrahydrofuran (400mL), stir to dissolve, and cool to At -20°C, sodium tert-butoxide (4.4 g, 46.0 mmol) was slowly added in batches. After the addition, the temperature was raised to room temperature and stirred overnight. After the reaction, the temperature was lowered to -10°C, water (60 mL) was slowly added dropwise to quench, ethyl acetate (100 mL) was added for extraction, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column (Petroleum Ether: ethyl acetate = 80:1 to 30:1) to obtain 2-bromo-3-fluoro-6-(vinyloxy)benzaldehyde (C1-c) (5g, 66% yield) as a pale yellow solid .

¹ H NMR (400MHz, CDCl ₃ ) δ 10.32 (d, J = 9.8 Hz, 1H), 7.29 (dt, J = 16.2, 8.3 Hz, 1H), 7.15-6.98 (m, 1H), 6.68-6.46 ( m, 1H), 4.89-4.68 (m, 1H), 4.68-4.42 (m, 1H) ppm.

Step 3: Intermediate (E)-N'-(2-bromo-3-fluoro-6-(vinyloxy)benzylidene)-4-methylbenzenesulfonyl hydrazide (C1-d):

Add 2-bromo-3-fluoro-6-(vinyloxy)benzaldehyde (C1-c) (5g, 20.4mmol) into a 250mL single-necked flask, add anhydrous methanol (100mL) and stir to dissolve, slowly add the pair at room temperature Tosyl hydrazide (4.2 g, 22.4 mmol) was stirred at room temperature for 18 hours. After the reaction, a large amount of white solid precipitated out. The temperature was lowered to 0°C. The solid product was collected by filtration. The mother liquor was concentrated. The crude solid product was slurried with a mixed solvent of petroleum ether and ethyl acetate (20:1). The solid product was collected and dried under reduced pressure to obtain (E )-N'-(2-Bromo-3-fluoro-6-(vinyloxy)benzylidene)-4-methylbenzenesulfonyl hydrazide (C1-d) (7.5g, 89% yield), White solid.

¹ H NMR(400MHz,DMSO-d ₆ )δ11.81(s,1H),7.97(s,1H),7.76(d,J=7.6Hz,2H),7.42(t,J=9.0Hz,2H) ,7.16(dd,J=9.1,4.3Hz,1H), 6.67(dd,J=13.6,6.2Hz,1H), 4.49(dd,J=21.4,9.8Hz,3H), 2.35(s,3H)ppm ; LCMS: m/z 414.1[M+H] ⁺ .

Step 4: Intermediate 6-bromo-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran (C1-e):

Add (E)-N'-(2-bromo-3-fluoro-6-(vinyloxy)benzylidene)-4-methylbenzenesulfonyl hydrazide (C1-d) (3g, 7.3mmol), add toluene (100mL) and stir to dissolve, slowly add lithium tert-butoxide (639mg, 7.9mmol) at room temperature, zinc rhodium acid dimer (56mg, 72.6μmmol) under nitrogen protection, warm to 100℃ and stir for 3 hours . After the reaction, the temperature was lowered to room temperature, filtered, the filter cake was washed with ethyl acetate (50 mL), and the filtrate was concentrated under reduced pressure to obtain 6-bromo-5-fluoro-1a,6b-dihydro-1H-cyclopropene[b] Benzofuran (C1-e) (1.67 g, 100% yield), as a pale yellow oil, was directly put into the next reaction without further purification.

¹ H NMR(400MHz, CDCl ₃ )δ7.03-6.81(m,2H), 4.97(t,J=5.2Hz,1H), 2.84(dt,J=9.0,4.4Hz,1H), 1.26(dt, J = 8.0, 6.1Hz, 1H), 0.55-0.37 (m, 1H) ppm.

Step 5: Intermediate 5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-carbonitrile (C1-f):

Add 6-bromo-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran (C1-e) (1.9g, 8.30mmol), zinc cyanide (1.46) to a 100mL single-mouth flask g, 12.4mmol), tetrakistriphenylphosphine palladium (1.44g, 1.24mmol), DMF (12mL), protected by nitrogen, heated to 110°C and stirred for 18 hours. After the reaction, the temperature was lowered to room temperature, ethyl acetate (20mL) was added to dilute, filtered, the filtrate was washed twice with saturated brine (10mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was passed through a silica gel column Purification (petroleum ether: ethyl acetate = 100:1 to 20:1) to obtain 5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-carbonitrile (C1-f) (1.35g, 92% yield), white solid.

¹ H NMR (400MHz, CDCl ₃ ) δ7.03-6.84 (m, 2H), 4.97 (t, J = 5.2 Hz, 1H), 2.84 (dt, J = 9.0, 4.5 Hz, 1H), 1.33-1.20 ( m, 1H), 0.52-0.40 (m, 1H) ppm.

Step 6: Intermediate tert-butyl ((5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)carbamate (C1-g) :

Add 5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-carbonitrile (C1-f) (1.2g, 6.8mmol) into a 100mL single-neck flask, and add methanol (30mL ) Dissolve, add di-tert-butyl dicarbonate (2.2g, 10.3mmol) at room temperature, cool to -5°C, slowly add nickel chloride hexahydrate (1.95g, 8.22mmol), add sodium borohydride in batches The solid (778mg, 20.5mmol) was naturally warmed to room temperature and stirred for 2 hours. After the reaction, filter with celite, wash the filter cake with methanol (10mL), cool the filtrate to 0℃, quench with water (5mL), concentrate under reduced pressure, add ethyl acetate (80mL) to the crude product, and use saturated salt for the organic phase Washed with water (10 mL), the organic phase was dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate = 100:1 to 40:1) to obtain tert-butyl (( 5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)carbamate (C1-g) (1.2g, 63% yield), White solid.

¹ H NMR (400MHz, CDCl ₃ ) δ 6.76 (t, J = 9.3 Hz, 1H), 6.64 (dd, J = 8.6, 3.7 Hz, 1H), 4.90 (s, 1H), 4.82 (t, J = 5.3Hz, 1H), 4.48 (d, J = 6.1Hz, 1H), 4.46-4.33 (m, 1H), 2.85 (d, J = 3.8 Hz, 1H), 1.44 (s, 9H), 1.05 (dd, J = 14.6, 6.0 Hz, 1H), 0.32 (s, 1H) ppm; LCMS: m/z224.1[M-55] ⁺ .

Step 7: Intermediate tert-butyl ((5-fluoro-1aS,6bS-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)carbamate (C1-h) And ((5-Fluoro-1aR,6bR-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)carbamate (C1-i):

The tert-butyl ((5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)carbamate (C1-g) (1.2g, 4.3mmol) was further purified by the following chiral SFC: Column: AD-H 20x250mm, 10um (Daicel), flow rate: 80g/min, mobile phase: 13% (0.2% ammonia methanol/methanol) in CO2, detection: 214nM to give tert-butyl ((5-fluoro-1aS,6bS-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)carbamate (C1-h)(0.57 g, 47.5% yield), Rt: 0.71min; and ((5-fluoro-1aR,6bR-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)carbamate (C1-i) (0.52g, 43.3% yield), Rt: 0.99min.

C1-h: ¹ H NMR (400MHz, CDCl ₃ ) δ 6.76 (t, J = 9.3 Hz, 1H), 6.64 (dd, J = 8.6, 3.9 Hz, 1H), 4.98-4.73 (m, 2H), 4.45(ddd,J=41.3,14.0,5.7Hz,2H),2.84(s,1H),1.45(d,J=7.7Hz,9H),1.05(dd,J=14.8,6.0Hz,1H),0.31 (d,J=4.6Hz,1H) ppm; LCMS: m/z224.1[M-55] ⁺ .

C1-i: ¹ H NMR (400MHz, CDCl ₃ ) δ 6.76 (t, J = 9.3 Hz, 1H), 6.64 (dd, J = 8.6, 3.9 Hz, 1H), 4.98-4.73 (m, 2H), 4.45(ddd,J=41.3,14.0,5.7Hz,2H),2.84(s,1H),1.45(d,J=7.7Hz,9H),1.05(dd,J=14.8,6.0Hz,1H),0.31 (d,J=4.6Hz,1H) ppm; LCMS: m/z224.1[M-55] ⁺ .

Step 8: Intermediate (5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methylamine (C)

Add the compound tert-butyl ((5-fluoro-1a, 6b-dihydro-1H-cyclopropyl[b]benzofuran-6-yl)methyl) tert-butyl carbamate (C1-g ) (1.2g, 4.30mmol), add dichloromethane (20mL) to dissolve, add trifluoroacetic acid (4mL, 52mmol) at room temperature, and stir for 2 hours. After the reaction is over, concentrate under reduced pressure to remove dichloromethane and trifluoroacetic acid A colorless solid (5-fluoro-1a, 6b-dihydro-1H-cyclopropyl[b]benzofuran-6-yl)methylamine (C1) trifluoroacetic acid hydrochloride was obtained, 1.26g, yield The rate is 100%. The product can be used directly in the next step.

¹ H NMR (400MHz, MeOD) δ 6.95 (t, J = 9.5 Hz, 1H), 6.84 (dd, J = 8.8, 4.0 Hz, 1H), 4.97 (t, J = 5.4 Hz, 1H), 4.31 ( q,J=13.7Hz,2H),2.92-2.77(m,1H),1.26-1.11(m,1H),0.38-0.21(m,1H)ppm; LCMS: m/z180.1[M+H] ⁺ .

((1aS,6bS)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6 was obtained by the method of step 8 using intermediates C1-h and C1-i respectively as raw materials -Yl)methylamine (C2) and ((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropen[b]benzofuran-6-yl)methylamine (C3)

Example 4: 8-Bromo-N-((5-fluoro-1a,6bdihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-[1,2,4]tri Azolo[4,3-C]pyrimidin-5-amine (A-1)

The compound (5-fluoro-1a, 6b-dihydro-1H-cyclopropyl (b) benzofuran-6-yl) methylamine (C1) trifluoroacetate (1.0g, 3.41mmol), 8-bromo-5-chloro-[1,2,4]triazolo[4,3-c]pyrimidine (B1) (0.95g, 4.09mmol), triethylamine (0.69g, 6.82mmol) ), acetonitrile (20mL), stirred at room temperature for 3 hours, after the reaction, add ethyl acetate (80mL) to dilute and dissolve, the organic phase was washed with saturated brine (10mL*2), the organic phase was dried over anhydrous sodium sulfate, and reduced pressure Concentrated and purified by silica gel column chromatography (mobile phase: PE:EA=30:1 to PE:EA=10:1) to obtain compound 8-bromo-N-((5-fluoro-1a, 6b dihydro-1H- Cyclopropeno[b]benzofuran-6-yl)methyl)-[1,2,4]triazolo[4,3-C]pyrimidin-5-amine (A-1) (1.2g, yield Rate 94%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.50 (s, 1H), 8.87 (d, J = 4.5 Hz, 1H), 7.85 (s, 1H), 6.94 (t, J = 9.5 Hz, 1H) ,6.77(dd,J=8.7,3.7Hz,1H), 4.92(t,J=5.2Hz,1H), 4.79(p,J=9.7Hz,1H), 2.88(dt,J=8.7,4.5Hz, 1H),0.97(dd,J=14.4,5.7Hz,1H),0.10(s,1H)ppm; LCMS: m/z376.0[M+H] ⁺ .

Using the method of Example 4, B1 was reacted with C2 and C3 to obtain intermediates A-2 and A-3.

Example 5:

The compound ((1aS,6bS)-5-fluoro-1a,6b-dihydro-1H-cyclopropen[b]benzofuran-6-yl)methylamine (C2) in a 100mL single-mouth flask Acid salt (1.0g, 3.41mmol), 5-chloro-8-iodopyrido[4,3-d]pyrimidine (B2) (1.2g, 4.09mmol), triethylamine (0.87g, 8.53mmol), acetonitrile (20mL), warm up to 75℃ and react for 2-3h. After the reaction, cool to room temperature, add ethyl acetate (100mL) to dilute and dissolve, wash the organic phase with saturated brine (20mL*2), and wash the organic phase with anhydrous sulfuric acid After drying with sodium and concentrating under reduced pressure, the crude product was purified by silica gel column chromatography to obtain the product N-((5-fluoro-1a,6b dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)- 8-Iodopyrido[4,3-d]pyrimidin-5-amine (A-4) (1.2 g, yield 81%).

¹ H NMR (400MHz, CDCl ₃ ) δ9.42 (s, 1H), 9.20 (s, 1H), 8.75 (s, 1H), 6.81 (t, J = 9.4 Hz, 1H), 6.78-6.60 (m, 1H), 6.12 (s, 1H), 5.04 (dd, J = 14.2, 5.9 Hz, 1H), 4.95-4.81 (m, 2H), 2.97-2.84 (m, 1H), 1.07 (dt, J = 17.0, 8.5Hz,1H),0.45-0.26(m,1H)ppm; LCMS: m/z435.0[M+H] ⁺ .

Example 6: N-(((1aS,6bS)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2- (Methylpyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

Add the compound 8-bromo-N-(((1aS,6bS)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methane in a 25mL single-necked flask in sequence Yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine (A-2) (50mg, 0.13mmol), (2-methylpyridin-3-yl)boronic acid Hydrochloride 35mg, 0.20mmol), potassium carbonate (36mg, 0.266mmol), [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride (15mg, 0.02mmol), 1,4 -Dioxane (4mL), H ₂ O (1mL), the reaction was heated to 80°C and reacted for 2h under nitrogen atmosphere. After the reaction, the temperature was lowered to room temperature, and ethyl acetate (20mL) was added to dilute and dissolve. The organic phase was diluted with water ( 5mL*2), the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate = 1:1) to obtain compound 8-bromo-N-(( (1aS,6bS)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-[1,2,4]triazolo[4, 3-c] Pyrimidine-5-amine (25 mg, yield 48%).

¹ H NMR(400MHz,CDCl ₃ )δ8.93(s,1H), 8.54(d,J=4.8Hz,1H), 7.72(d,J=7.6Hz,1H), 7.63(s,1H), 7.22 (dd,J=7.6,5.0Hz,1H), 6.80(t,J=9.4Hz,1H), 6.69(dd,J=8.7,3.9Hz,1H), 6.46(t,J=5.3Hz,1H) ,5.01(dd,J=14.2,5.8Hz,1H),4.97-4.82(m,2H),2.96(dt,J=9.0,4.6Hz,1H),2.54(s,3H),1.08(dd,J = 14.8, 6.0 Hz, 1H), 0.34 (s, 1H) ppm; LCMS: m/z389.1[M+H] ⁺ .

Using the method of Example 6, the following compounds can be synthesized:

Example 7: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2- (Methylpyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,CDCl ₃ )δ8.93(s,1H), 8.54(d,J=4.8Hz,1H), 7.72(d,J=7.6Hz,1H), 7.63(s,1H), 7.22 (dd,J=7.6,5.0Hz,1H), 6.80(t,J=9.4Hz,1H), 6.69(dd,J=8.7,3.9Hz,1H), 6.46(t,J=5.3Hz,1H) ,5.01(dd,J=14.2,5.8Hz,1H),4.97-4.82(m,2H),2.96(dt,J=9.0,4.6Hz,1H),2.54(s,3H),1.08(dd,J = 14.8, 6.0 Hz, 1H), 0.34 (s, 1H) ppm; LCMS: m/z389.1[M+H] ⁺ .

Example 8: N-(((1a,6b)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2- (Methylpyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,CDCl ₃ )δ9.05(s,1H), 8.54(d,J=4.5Hz,1H), 7.72(d,J=7.6Hz,1H), 7.62(s,1H), 7.19 -7.14(m,1H),6.79(t,J=9.3Hz,2H),6.68(dd,J=8.7,3.9Hz,1H),5.08-4.97(m,1H),4.97-4.75(m,2H ),3.00-2.93(m,1H),2.54(s,3H),0.92-0.42(m,1H),0.33(s,1H)ppm; LCMS: m/z389.1[M+H] ⁺ .

Example 9: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(4-(trifluoromethyl) Phenyl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.41(s,1H), 8.87(t,J=4.9Hz,1H), 8.27(d,J=8.2Hz,2H), 8.09(s,1H) ,7.69(d,J=8.2Hz,2H), 6.84(dd,J=22.7,13.4Hz,1H), 6.65(dd,J=8.7,3.7Hz,1H), 4.89-4.69(m,3H), 2.80(dt,J＝9.0,4.5Hz,1H),0.86(dt,J＝8.8,5.8Hz,1H),0.01(s,1H)ppm; LCMS: m/z442.0[M+H] ⁺ .

Example 10: 8-(3,6-Dihydro-2H-pyran-4-yl)-N-((5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran -6-yl)methyl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.47(s,1H), 8.75(t,J=4.9Hz,1H), 7.67(s,1H),7.35(s,1H), 6.93(t, J = 9.5Hz, 1H), 6.76 (dd, J = 8.7, 3.7 Hz, 1H), 4.91 (dd, J = 14.4, 9.3 Hz, 1H), 4.89-4.77 (m, 2H), 4.29 (s, 2H) ), 3.85 (t, J = 5.4 Hz, 2H), 2.89 (dt, J = 8.9, 4.5 Hz, 1H), 2.51 (s, 2H), 0.95 (dd, J = 14.5, 5.9 Hz, 1H), 0.08 (s,1H) ppm; LCMS: m/z380.1[M+H] ⁺ .

Example 11: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno(b)benzofuran-6-yl)methyl)-8-(2-fluorophenyl)-[ 1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.37(s,1H), 8.78(d,J=4.5Hz,1H), 7.72(dd,J=17.7,9.7Hz,2H), 7.32(dd, J = 13.4, 6.7 Hz, 1H), 7.20 (dd, J = 15.7, 8.3 Hz, 2H), 6.82 (t, J = 9.5 Hz, 1H), 6.65 (dd, J = 8.6, 3.7 Hz, 1H), 4.81(t,J=4.8Hz,1H), 4.73(p,J=9.9Hz,2H), 2.85-2.75(m,1H), 0.91-0.83(m,1H), 0.01(s,1H)ppm; LCMS: m/z392.0[M+H] ⁺ .

Example 12: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno(b)benzofuran-6-yl)methyl)-8-(pyrimidin-5-yl)-[ 1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.55(d,J=5.7Hz,3H), 9.15(s,1H), 9.10(s,1H), 8.31(s,1H), 6.95(t, J = 9.5Hz, 1H), 6.78 (dd, J = 8.7, 3.7 Hz, 1H), 4.94 (s, 1H), 4.89 (s, 2H), 2.96-2.90 (m, 1H), 0.99 (dd, J =14.6,5.8Hz,1H),0.13(s,1H)ppm; LCMS: m/z375.8[M+H] ⁺ .

Example 13: 4-(5-(((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)amino)-[1,2, 4] Triazolo[4,3-c]pyrimidin-8-yl)benzamide

¹ H NMR(400MHz,DMSO-d ₆ )δ9.54(s,1H),8.95(s,1H), 8.24(d,J=8.2Hz,2H), 8.18(s,1H), 8.02(s, 1H), 7.96 (d, J = 8.2 Hz, 2H), 7.38 (s, 1H), 6.95 (t, J = 9.5 Hz, 1H), 6.78 (dd, J = 8.6, 3.7 Hz, 1H), 5.03- 4.78 (m, 3H), 2.94 (dt, J = 8.9, 4.4 Hz, 1H), 0.98 (dd, J = 14.5, 5.8 Hz, 1H), 0.12 (s, 1H) ppm; LCMS: m/z 416.9 [M+H] ⁺ .

Example 14: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno(b)benzofuran-6-yl)methyl)-8-(4-methylpyridine-3- Yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR (400MHz, CDCl ₃ ) δ 8.91 (s, 1H), 8.55 (s, 1H), 8.51 (d, J = 4.8 Hz, 1H), 7.66 (s, 1H), 7.36 (d, J = 5.0Hz, 1H), 6.82 (t, J = 9.5 Hz, 1H), 6.71 (dd, J = 8.7, 3.9 Hz, 1H), 6.36 (s, 1H), 5.04 (d, J = 13.9 Hz, 1H) ,4.96-4.83(m,2H),2.96(d,J=4.2Hz,1H),2.32(s,3H),1.10(dd,J=14.4,6.2Hz,1H),0.36(s,1H)ppm ; LCMS: m/z388.9[M+H] ⁺ .

Example 15: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(6-fluoro-2-methyl (Pyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,CDCl ₃ )δ8.92(s,1H),7.83(t,J=8.0Hz,1H),7.62(s,1H),6.83(dd,J=17.2,8.0Hz,2H) ,6.71(dd,J=8.6,3.5Hz,1H), 6.24(s,1H), 5.03(d,J=10.1Hz,1H), 4.89(d,J=22.4Hz,2H), 2.97(s, 1H),2.48(s,3H),0.86(d,J=6.9Hz,1H),0.36(s,1H)ppm; LCMS: m/z 407.1[M+H] ⁺ .

Example 16: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(4-fluorophenyl)-[ 1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR (400MHz, CDCl ₃ ) δ 8.81 (s, 1H), 8.01-7.95 (m, 2H), 7.87 (s, 1H), 7.17 (t, J = 8.4 Hz, 2H), 6.80 (d, J = 9.6Hz, 1H), 6.71 (d, J = 3.8Hz, 1H), 5.79 (s, 1H), 5.03 (s, 1H), 4.86 (s, 2H), 2.99-2.91 (m, 1H), 1.10(dd,J=14.6,6.2Hz,1H),0.36(s,1H)ppm; LCMS: m/z392.12[M+H] ⁺ .

Example 17: 4-(5-(((5-Fluoro-1a,6b-dihydro-1H-cyclopropyl[b]benzofuran-6-yl)methyl)amino)-[1,2, 4]Triazolo[4,3]tert-butyl-c]pyrimidin-8-yl)-5,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester

¹ H NMR(400MHz,DMSO-d ₆ )δ9.46(s,1H), 8.76(t,J=4.8Hz,1H), 7.68(s,1H), 7.25(s,1H), 6.93(t, J = 9.5Hz, 1H), 6.76 (dd, J = 8.6, 3.6Hz, 1H), 4.92 (t, J = 5.1Hz, 1H), 4.87-4.76 (m, 2H), 4.07 (s, 2H), 3.56(s,2H),2.89(dt,J=8.8,4.4Hz,1H),2.54(s,2H),1.43(s,9H),0.96-0.91(m,1H),0.08(s,1H) ppm; LCMS: m/z478.9[M+H] ⁺ .

Example 18: 8-(3,5-Dimethylisothiazol-4-yl)-N-((5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6 -Yl)methyl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR (400MHz, DMSO-d ₆ ) δ9.48 (s, 1H), 8.87 (t, J = 4.7Hz, 1H), 7.67 (s, 1H), 6.99-6.92 (m, 1H), 6.78 ( dd,J=8.7,3.8Hz,1H), 4.94(t,J=5.1Hz,1H), 4.89-4.79(m,2H), 2.94(dt,J=8.9,4.4Hz,1H), 2.37(s ,3H), 2.19(s,3H),0.98(dd,J=14.6,5.9Hz,1H),0.12(s,1H)ppm; LCMS: m/z392.8[M+H] ⁺ .

Example 19: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2-fluoropyridin-4-yl )-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.56(d,J=1.2Hz,1H), 9.23(s,1H), 8.51(d,J=1.2Hz,1H), 8.29(d,J= 5.4Hz, 1H), 8.26-8.19 (m, 1H), 8.10 (s, 1H), 6.96 (t, J = 9.5 Hz, 1H), 6.79 (dd, J = 8.8, 3.9 Hz, 1H), 5.06- 4.80(m,3H),2.92(dt,J=9.2,4.6Hz,1H),0.98(dd,J=9.2,5.4Hz,1H),0.12(ddd,J=5.9,3.9,1.8Hz,1H) ppm; LCMS: m/z393.1[M+H] ⁺ .

Example 20: 8-(2,4-Difluorophenyl)-N-((5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl )-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.50 (s, 1H), 8.92 (s, 1H), 7.91 (q, J = 8.3 Hz, 1H), 7.80 (s, 1H), 7.49-7.33 ( m, 1H), 7.30-7.14 (m, 1H), 6.95 (t, J = 9.5 Hz, 1H), 6.78 (dd, J = 8.7, 3.9 Hz, 1H), 5.05-4.75 (m, 3H), 2.93 (dt,J=9.2,4.6Hz,1H),1.09-0.86(m,1H),0.13(t,J=5.0Hz,1H)ppm; LCMS410.1[M+H] ⁺ .

Example 21: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(4-(methylsulfonyl)benzene Yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.55(s,1H),9.06(s,1H),8.45(d,J=8.2Hz,2H),8.26(s,1H),8.00(d, J = 8.2Hz, 2H), 6.96 (t, J = 9.5Hz, 1H), 6.79 (dd, J = 8.6, 3.8Hz, 1H), 5.06-4.80 (m, 3H), 3.26 (s, 3H), 2.94(dt,J=9.1,4.6Hz,1H),0.99(dd,J=9.7,5.1Hz,1H),0.13(t,J=5.1Hz,1H)ppm; LCMS452.1[M+H] ⁺ .

Example 22: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(4-methoxyphenyl)- [1,2,4]Triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.38(s,1H),8.63(s,1H),7.93(d,J=8.5Hz,2H),7.85(s,1H),6.93(d, J = 8.4Hz, 2H), 6.83 (t, J = 9.5 Hz, 1H), 6.66 (dd, J = 8.7, 3.8 Hz, 1H), 4.81 (t, J = 5.0 Hz, 1H), 4.74 (d, J = 3.8 Hz, 2H), 2.81 (dt, J = 9.1, 4.4 Hz, 1H), 1.89 (q, J = 7.8 Hz, 1H), 0.86 (dd, J = 9.3, 5.3 Hz, 1H), 0.11 ( s,1H) ppm; LCMS: m/z404.1[M+H] ⁺ .

Example 23: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2-morpholinopyrimidine-5 -Yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.50(s,1H), 9.09(d,J=3.9Hz,2H), 8.83(t,J=4.9Hz,1H), 8.03(s,1H) ,6.94(t,J=9.5Hz,1H), 6.77(dd,J=8.6,3.7Hz,1H), 4.93(t,J=4.9Hz,1H), 4.85(t,J=4.1Hz,2H) ,3.76(d,J＝4.4Hz,4H), 3.69(d,J＝4.0Hz,4H), 2.91(dt,J＝9.0,4.5Hz,1H),0.98(dd,J＝14.6,5.8Hz, 1H),0.12(s,1H)ppm; LCMS: m/z461.18[M+H] ⁺ .

Example 24: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2- (Trifluoromethyl)pyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.55(s,1H),9.06(s,1H),8.83(d,J=4.3Hz,1H), 8.09(d,J=7.7Hz,1H) ,7.84(dd,J=7.8,4.7Hz,1H),7.62(d,J=10.9Hz,1H),7.01-6.83(m,1H),6.77(dt,J=11.1,5.6Hz,1H), 5.01-4.75(m,3H),3.02-2.92(m,1H),1.03-0.81(m,1H),0.14-0.03(m,1H)ppm; LCMS: m/z442.8[M+H] ⁺ .

Example 25: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2- Methoxypyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.49(s,1H),8.86(s,1H),8.30-8.13(m,2H),7.95(s,1H),7.14(d,J=5.0 Hz, 1H), 7.00-6.90 (m, 1H), 6.79 (s, 1H), 5.05-4.76 (m, 3H), 3.87 (s, 3H), 3.00-2.87 (m, 1H), 2.07-1.94 ( m,1H),1.04-0.94(m,1H),0.23-0.10(m,1H)ppm; LCMS: m/z404.8[M+H] ⁺ .

Example 26: 8-(1,3-Dimethyl-1H-pyrazol-5-yl)-N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropene And [b]benzofuran-6-yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-5-amine

¹ H NMR(400MHz, CDCl ₃ )δ9.35(s,1H), 8.20(s,1H), 7.63(s,1H), 6.55(t,J=9.3Hz,1H), 6.48(dd,J= 8.5, 3.7 Hz, 1H), 6.19 (s, 1H), 4.78 (d, J = 4.9 Hz, 1H), 4.70 (dd, J = 12.4, 5.4 Hz, 2H), 3.72 (s, 3H), 2.91 2.78(m,1H), 2.16(s,3H), 0.91(d,J=8.2Hz,1H), 0.17(s,1H)ppm; LCMS: m/z319.8[M+H] ⁺ . Example 27: 8-(1,3-Dimethyl-1H-pyrazol-4-yl)-N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[ b]benzofuran-6-yl)methyl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.47(s,1H), 8.64(t,J=4.8Hz,1H), 8.28(s,1H), 7.73(s,1H), 6.94(t, J=9.5Hz,1H), 6.77(dd,J=8.7,3.8Hz,1H), 4.93(t,J=4.9Hz,1H), 4.88-4.78(m,2H), 3.83(s,3H), 2.95-2.88(m,1H),2.33(s,3H),0.96(dd,J=14.6,5.8Hz,1H),0.13-0.08(m,1H)ppm; LCMS: m/z391.9[M+ H] ⁺ .

Example 28: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2- (Methylpyridin-4-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.54(s,1H), 9.08(t,J=5.2Hz,1H), 8.48(d,J=5.3Hz,1H), 8.33(s,1H) ,8.11(s,1H),8.00(d,J=5.3Hz,1H),7.00-6.91(m,1H), 6.78(dd,J=8.8,3.9Hz,1H), 4.91(dt,J=16.1 ,3.8Hz,3H),2.96-2.89(m,1H),2.53(s,3H),1.02-0.94(m,1H),0.14-0.09(m,1H)ppm; LCMS: m/z388.9[ M+H] ⁺ .

Example 29: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(pyrimidine- 5-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.55(d,J=6.4Hz,3H), 9.15(s,1H), 9.10(s,1H), 8.31(s,1H), 6.95(t, J=9.5Hz,1H), 6.78(dd,J=8.7,3.7Hz,1H), 4.93(dd,J=14.2,9.3Hz,3H), 2.92(dt,J=9.1,4.6Hz,1H), 0.99(dd,J=14.6,5.8Hz,1H),0.13(s,1H)ppm; LCMS: m/z376.13[M+H] ⁺ .

Example 30: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(6- Fluoro-2-methylpyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.50(s,1H), 8.89(t,J=4.7Hz,1H), 7.96(t,J=8.2Hz,1H), 7.66(s,1H) ,7.15-7.08(m,1H),6.95(t,J=9.5Hz,1H), 6.78(dd,J=8.6,3.7Hz,1H), 4.94(t,J=4.9Hz,1H), 4.86( t,J=5.0Hz,2H),2.94(dt,J=8.9,4.4Hz,1H),2.35(s,3H),0.98(dd,J=14.3,5.7Hz,1H),0.12(s,1H ) ppm; LCMS: m/z407.1[M+H] ⁺ . Example 31: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b] Benzofuran-6-yl)methyl)-8-(6-methoxypyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.51(s,1H),8.88(s,1H),8.84(s,1H), 8.41(d,J=8.5Hz,1H), 8.04(s, 1H), 6.95 (t, J = 8.2 Hz, 2H), 6.78 (dd, J = 8.7, 3.8 Hz, 1H), 4.93 (t, J = 4.9 Hz, 1H), 4.84 (d, J = 14.8 Hz, 2H),3.90(s,3H),2.96-2.88(m,1H),0.98(dd,J=14.5,5.9Hz,1H),0.12(s,1H)ppm; LCMS: m/z405.14[M +H] ⁺ .

Example 32: 4-(5-((((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)amino) -[1,2,4]triazolo[4,3-c]pyrimidin-8-yl)benzamide

¹ H NMR(400MHz,DMSO-d ₆ )δ9.41(s,1H),8.81(s,1H),8.22-7.99(m,3H),7.96-7.74(m,3H),7.27(s,1H) ), 6.83(t,J=9.4Hz,1H), 6.70-6.61(m,1H), 4.87-4.61(m,3H), 2.88-2.74(m,1H), 0.93-0.81(m,1H), 0.06--0.06(m,1H)ppm; LCMS: m/z416.8[M+H] ⁺ .

Example 33: 8-(6-chloropyridin-3-yl)-N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran- 6-yl)methyl)-[1,2,4]triazolo[4,-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.45(s,1H), 9.06(s,1H), 8.98(s,1H), 8.52(d,J=8.8Hz,1H), 8.12(s, 1H), 7.52 (d, J = 8.6 Hz, 1H), 6.83 (t, J = 9.2 Hz, 1H), 6.67 (d, J = 8.2 Hz, 1H), 4.87-4.71 (m, 3H), 2.86 2.81(m,1H),0.90-0.83(m,1H),0.03--0.03(m,1H)ppm; LCMS: m/z408.7[M+H] ⁺ .

Example 34: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(pyridine- 3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.45(s,1H), 9.06(s,1H), 8.98(s,1H), 8.52(d,J=8.8Hz,1H), 8.12(s, 1H), 7.52 (d, J = 8.6 Hz, 1H), 6.83 (t, J = 9.2 Hz, 1H), 6.67 (d, J = 8.2 Hz, 1H), 4.87-4.71 (m, 3H), 2.86 2.81(m,1H),0.90-0.83(m,1H),0.03--0.03(m,1H)ppm; LCMS: m/z408.7[M+H] ⁺ .

Example 35: 8-(2-chloropyridin-3-yl)-N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran- 6-yl)methyl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.51(s,1H),9.00(s,1H),8.48(d,J=4.5Hz,1H), 8.05(d,J=7.5Hz,1H) ,7.76(s,1H),7.61-7.52(m,1H),6.99-6.92(m,1H),6.78(dd,J=8.4,4.0Hz,1H),4.99-4.78(m,3H),2.96 -2.88(m,1H),0.99(t,J=7.4Hz,1H),0.12(s,1H)ppm; LCMS: m/z408.08[M+H] ⁺ .

Example 36: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2- (Fluoropyridin-3-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.52(s,1H),9.02(s,1H),8.53(t,J=8.7Hz,1H), 8.26(d,J=4.3Hz,1H) ,7.96(s,1H),7.59-7.45(m,1H),6.95(t,J=9.5Hz,1H),6.79(dd,J=8.7,3.8Hz,1H),5.16-4.69(m,3H ), 2.94(dt,J=8.7,4.5Hz,1H),1.00(dd,J=14.6,5.8Hz,1H),0.13(s,1H)ppm; LCMS: m/z392.8[M+H] ⁺ .

Example 37: 8-(2-Aminopyridin-3-yl)-N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran- 6-yl)methyl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.46(s,1H),8.76(s,1H),8.00(d,J=4.9Hz,1H),7.60(s,1H),7.45(d, J=7.2Hz,1H),7.04-6.91(m,1H),6.78(dd,J=8.5,3.6Hz,1H), 6.72-6.60(m,1H),5.72(s,2H), 4.95(t ,J = 5.1Hz, 1H), 4.84 (s, 2H), 3.01-2.90 (m, 1H), 1.03 (dd, J = 14.7, 6.0 Hz, 1H), 0.17 (s, 1H) ppm; LCMS: m /z389.8[M+H] ⁺ .

Example 38: (3-(5-((((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)amino )-[1,2,4]triazolo[4,3-c]pyrimidin-8-yl)pyridin-2-yl)methanol

¹ H NMR(400MHz,DMSO-d ₆ )δ9.50(s,1H),8.89(s,1H),8.61(d,J=4.8Hz,1H),7.89(d,J=7.6Hz,1H) ,7.75(s,1H),7.44(dd,J=7.7,4.8Hz,1H),7.00-6.93(m,1H), 6.79(dd,J=8.6,3.7Hz,1H), 5.13(t,J =5.7Hz,1H),4.97-4.83(m,3H),4.50(d,J=5.7Hz,2H),2.95(dt,J=9.2,4.7Hz,1H),0.99(dd,J=14.5, 5.9Hz,1H),0.17-0.11(m,1H)ppm; LCMS: m/z404.8[M+H] ⁺ .

Example 39: 8-(2,3-Dichloropyridin-4-yl)-N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzene And furan-6-yl)methyl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.52(s,1H), 9.16(s,1H), 8.48(d,J=5.0Hz,1H), 7.89(s,1H), 7.71(d, J = 4.6Hz, 1H), 6.99-6.92 (m, 1H), 6.79 (dd, J = 8.6, 3.8Hz, 1H), 4.97-4.92 (m, 1H), 4.88 (d, J = 9.1Hz, 2H ), 2.97-2.91(m,1H),0.99(dd,J=14.7,6.0Hz,1H),0.13(s,1H)ppm; LCMS: m/z443.26[M+H] ⁺ .

Example 40: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(1- Methyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.50 (s, 1H), 8.87 (t, J = 4.7 Hz, 1H), 8.46 (s, 1H), 7.65 (s, 1H), 6.95 (t, J = 9.4 Hz, 1H), 6.78 (dd, J = 8.7, 3.6 Hz, 1H), 4.93 (t, J = 5.2 Hz, 1H), 4.84 (qd, J = 14.7, 5.0 Hz, 2H), 4.01 ( s,3H), 2.91(dt,J=8.7,4.5Hz,1H),0.99-0.92(m,1H),0.09(s,1H)ppm; LCMS: m/z446.13[M+H] ⁺ .

Example 41: 8-(3,5-Dimethylisothiazol-4-yl)-N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b ]Benzofuran-6-yl)methyl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR (400MHz, DMSO-d ₆ ) δ9.48 (s, 1H), 8.87 (t, J = 4.7Hz, 1H), 7.67 (s, 1H), 6.99-6.92 (m, 1H), 6.78 ( dd,J=8.7,3.8Hz,1H), 4.94(t,J=5.1Hz,1H), 4.89-4.79(m,2H), 2.94(dt,J=8.9,4.4Hz,1H), 2.37(s ,3H), 2.19(s,3H),0.98(dd,J=14.6,5.9Hz,1H),0.12(s,1H)ppm; LCMS: m/z392.8[M+H] ⁺ .

Example: 42: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2 -Morpholinopyrimidin-5-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.50(s,1H), 9.09(d,J=3.9Hz,2H), 8.83(t,J=4.9Hz,1H), 8.03(s,1H) ,6.94(t,J=9.5Hz,1H), 6.77(dd,J=8.6,3.7Hz,1H), 4.93(t,J=4.9Hz,1H), 4.85(t,J=4.1Hz,2H) ,3.76(d,J＝4.4Hz,4H), 3.69(d,J＝4.0Hz,4H), 2.91(dt,J＝9.0,4.5Hz,1H),0.98(dd,J＝14.6,5.8Hz, 1H),0.12(s,1H)ppm; LCMS: m/z461.18[M+H] ⁺ .

Example 43: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(4- (Methanesulfonyl)phenyl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.55(s,1H),9.06(s,1H),8.45(d,J=8.2Hz,2H),8.26(s,1H),8.00(d, J = 8.2Hz, 2H), 6.96 (t, J = 9.5Hz, 1H), 6.79 (dd, J = 8.6, 3.8Hz, 1H), 5.06-4.80 (m, 3H), 3.26 (s, 3H), 2.94(dt,J=9.1,4.6Hz,1H),0.99(dd,J=9.7,5.1Hz,1H),0.13(t,J=5.1Hz,1H)ppm; LCMS452.1[M+H] ⁺ .

Example 44: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(1,2,3,6- Tetrahydropyridin-4-yl)-[1,2,4]triazolo[4,3-c]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.43(s,1H), 8.83(t,J=4.7Hz,1H), 8.78(s,1H), 7.69(s,1H), 7.28(s, 1H), 6.87 (t, J = 9.5 Hz, 1H), 6.70 (dd, J = 8.7, 3.5 Hz, 1H), 4.85 (t, J = 5.1 Hz, 1H), 4.76 (s, 2H), 3.78 ( s, 2H), 3.30 (s, 2H), 2.83 (dt, J = 8.9, 4.5 Hz, 1H), 2.69 (s, 2H), 0.86 (dd, J = 14.5, 5.8 Hz, 1H), 0.01 (s ,1H) ppm; LCMS: m/z378.9[M+H] ⁺ .

Example 45: N-(((1aS,6bS)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2- (Methylpyridin-3-yl)pyrido[4,3-d]pyrimidin-5-amine

¹ H NMR(400MHz, CDCl ₃ )δ9.38(s,1H), 9.32(s,1H), 8.58(d,J=4.6Hz,1H), 8.31(s,1H), 7.57(d,J= 7.4Hz,1H),7.25-7.21(m,1H),6.90-6.82(m,1H),6.72(dd,J=8.8,4.1Hz,1H), 6.09(s,1H), 5.12(dd,J = 14.1, 5.7 Hz, 1H), 4.94 (dd, J = 14.1, 4.9 Hz, 1H), 4.87 (t, J = 5.5 Hz, 1H), 3.03-2.94 (m, 1H), 2.38 (s, 3H) ,1.10(dd,J=14.7,5.9Hz,1H),0.39(s,1H)ppm; LCMS: m/z400.1[M+H] ⁺ .

Example 46: N-(((1aR,6bR)-5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2- (Methylpyridin-3-yl)pyrido[4,3-d]pyrimidin-5-amine

¹ H NMR(400MHz, CDCl ₃ )δ9.38(s,1H), 9.32(s,1H), 8.58(d,J=4.6Hz,1H), 8.31(s,1H), 7.57(d,J= 7.4Hz,1H),7.25-7.21(m,1H),6.90-6.82(m,1H),6.72(dd,J=8.8,4.1Hz,1H), 6.09(s,1H), 5.12(dd,J = 14.1, 5.7 Hz, 1H), 4.94 (dd, J = 14.1, 4.9 Hz, 1H), 4.87 (t, J = 5.5 Hz, 1H), 3.03-2.94 (m, 1H), 2.38 (s, 3H) ,1.10(dd,J=14.7,5.9Hz,1H),0.39(s,1H)ppm; LCMS: m/z400.1[M+H] ⁺ .

Example 47: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(pyrimidin-5-yl)pyrido [4,3-d]pyrimidin-5-amine

¹ H NMR (400MHz, DMSO-d ₆ ) δ 9.83 (s, 1H), 9.24 (s, 1H), 9.04 (s, 1H), 9.01 (s, 2H), 8.88 (t, J = 4.9 Hz, 1H), 8.48 (d, J = 14.5 Hz, 1H), 6.81 (t, J = 9.5 Hz, 1H), 6.63 (dd, J = 8.7, 3.7 Hz, 1H), 4.79 (d, J = 4.8 Hz, 3H), 2.83 (dt, J = 8.9, 4.5 Hz, 1H), 0.85 (dd, J = 14.4, 5.8 Hz, 1H), 0.01 (s, 1H) ppm; LCMS: m/z386.9[M+H ] ⁺ .

Example 48: 4-(5-(((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)amino)pyrido[4,3 -d]pyrimidin-8-yl)benzamide

¹ H NMR(400MHz,DMSO-d ₆ )δ9.95(s,1H),9.32(s,1H),8.92(s,1H),8.45(s,1H),8.02(s,1H),7.93( d,J=7.9Hz,2H),7.73(d,J=7.9Hz,2H),7.37(s,1H),6.95-6.88(m,1H),6.74(dd,J=8.5,3.6Hz,1H ), 4.90 (t, J = 5.4 Hz, 3H), 2.97 (dt, J = 10.8, 5.5 Hz, 1H), 0.94 (dd, J = 14.3, 5.7 Hz, 1H), 0.09 (s, 1H) ppm; LCMS: m/z427.8[M+H] ⁺ .

Example 49: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(4-methylpyridine-3- Yl)pyrido[4,3-d]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.94(s,1H), 9.23(d,J=5.6Hz,1H), 8.94(s,1H), 8.49(s,1H), 8.43(d, J = 4.9 Hz, 1H), 8.24 (s, 1H), 7.25 (d, J = 4.9 Hz, 1H), 6.92 (t, J = 9.6 Hz, 1H), 6.74 (dd, J = 8.7, 3.8 Hz, 1H),4.93-4.86(m,3H),2.97(dt,J＝8.6,4.4Hz,1H),2.05(s,3H),0.91(dd,J＝14.5,5.8Hz,1H),0.07(s ,1H) ppm; LCMS: m/z399.9[M+H] ⁺ .

Example 50: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(6-fluoro-2-methyl (Pyridin-3-yl)pyrido[4,3-d]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.91(s,1H),9.25(s,1H),8.85(s,1H),8.27(s,1H),7.81(t,J=8.2Hz, 1H), 7.06(d,J=7.9Hz,1H), 6.96-6.89(m,1H), 6.78-6.72(m,1H), 4.96-4.84(m,3H), 2.99-2.92(m,1H) ,2.18(s,3H),0.93(dd,J=13.5,4.7Hz,1H),0.12-0.06(m,1H)ppm; LCMS: m/z417.8[M+H] ⁺ .

Example 51: 8-(3,5-Dimethylisothiazol-4-yl)-N-((5-fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6 -Yl)methyl)pyrido[4,3-d]pyrimidin-5-amine

¹ H NMR(400MHz, DMSO-d ₆ )δ9.90(s,1H), 9.28(s,1H), 8.85(d,J=4.9Hz,1H), 8.29(s,1H), 6.96-6.90( m, 1H), 6.74 (dd, J = 8.6, 3.8 Hz, 1H), 4.95-4.83 (m, 3H), 2.94 (dt, J = 9.2, 4.6 Hz, 1H), 2.26 (s, 3H), 2.06 (s,3H),0.93(dd,J=14.5,5.8Hz,1H),0.11-0.05(m,1H)ppm; LCMS: m/z403.8[M+H] ⁺ .

Example 52: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(2-fluoropyridin-4-yl )Pyrido[4,3-d]pyrimidin-5-amine

¹ H NMR(400MHz,DMSO-d ₆ )δ9.94(s,1H),9.37(s,1H),9.09(d,J=4.8Hz,1H),8.63(s,1H), 8.27(d, J = 5.2Hz, 1H), 7.76 (d, J = 5.0 Hz, 1H), 7.59 (s, 1H), 6.92 (t, J = 9.6 Hz, 1H), 6.74 (dd, J = 8.8, 3.6 Hz, 1H), 4.91 (d, J = 4.4 Hz, 3H), 2.94 (dt, J = 8.8, 4.4 Hz, 1H), 0.95 (dd, J = 14.5, 5.8 Hz, 1H), 0.11 (t, J = 6.8 Hz,1H) ppm; LCMS: m/z403.8[M+H] ⁺ .

Example 53: N-((5-Fluoro-1a,6b-dihydro-1H-cyclopropeno[b]benzofuran-6-yl)methyl)-8-(4-(methylsulfonyl)benzene Yl)pyrido[4,3-d]pyrimidin-5-amine

¹ H NMR(400MHz, CDCl ₃ )δ9.39(d,J=12.6Hz,2H), 8.50(s,1H), 8.05(d,J=8.0Hz,2H), 7.89(d,J=8.1Hz , 2H), 6.84 (t, J = 9.4 Hz, 1H), 6.70 (dd, J = 8.6, 3.8 Hz, 1H), 6.25 (s, 1H), 5.14 (dd, J = 14.2, 5.8 Hz, 1H) , 4.98-4.84 (m, 2H), 2.98 (dt, J = 8.7, 4.5 Hz, 1H), 1.10 (dd, J = 14.5, 6.1 Hz, 1H), 0.38 (s, 1H) ppm; LCMS: m/ z463.1[M+H] ⁺ .

Pharmacology and application

As one of the main components of the PRC2 protein complex, EED does not have enzymatic activity, but it plays an important role in the overall function of PRC2. The effect of EED on PRC2 is specifically manifested in two aspects: 1) EED directly binds to the trimethylated H3K27Me3, which can locate the PCR2 complex on the chromatin that needs to be modified; 2) EED has the enzymatic catalytic function of EZH2 Great allosteric promotion. Therefore, the development of target compounds as the allosteric protein EED provides a new strategy to inhibit EZH2 enzyme activity. Moreover, such inhibitors have better or complementary advantages than EZH2 enzyme catalytic site inhibitors. For example, when patients develop resistance to EZH2 enzyme inhibitors, EED inhibitors can also inhibit EZH2 enzyme activity. The present invention discloses that the compound can be used as an EED target inhibitor, and has a therapeutic effect on diseases related to the mechanism of EED and/or PRC2.

The biological function of the compound disclosed in the present invention has been proved in biochemical and cellular level tests. For example, in a biochemical test, the compound disclosed in the present invention can have a strong competitive binding effect with the H3K27Me3 polypeptide that binds to the EED protein (IC ₅₀ can reach <10 nM). At the cellular level, the compound disclosed in the present invention can not only inhibit the methylation level of histone H3K27, but also inhibit the proliferation of cancer cells through this effect.

Example 54: Evaluation of the compound's effect on blocking the binding of EED and H3K27me3 by AlphaScreen (a-screen) method

First configure compound solutions with different concentration gradients. The compound powder is dissolved in DMSO into a mother liquor. Take 1.5μl of compound mother solution to 198.5μl of reaction buffer (25mM HEPES (pH8.0), 50mM NaCl, 0.015% Tween20, 0.5% BSA) and mix well, and then use the above buffer containing 0.75% DMSO to perform a 3-fold gradient Dilution, set 9 different test concentrations for the same compound. Take 5μL of the compounds with different concentration gradients to ProxiPlate-384Plus, White detection plate (PerkinElmer, 6008280), and set 2 parallel replicates for each concentration gradient.

Secondly, the binding blocking reaction is performed. Dilute the full-length EED protein (441 amino acids) with His6 tag to 60 nM and the biotin-labeled polypeptide fragment H3K27me3 (amino acids 19-33) (Biotinylated-H3K27me3) to 75 nM with the above buffer. Transfer 5μl of 75nM polypeptide fragment and 5μl of 60nM protein to the detection well containing the compound, seal the detection plate with a film, and incubate at room temperature for 30 minutes.

Finally, the AlphaScreen method is tested. Immediately before use, the nickel chelated acceptor beads and the streptavidin donor beads were mixed in the above reaction buffer at a ratio of 1:1 (Perkin Elmer, product number 6760619M), and then 5μl of the above pretreatment was added to each detection well. The final concentration of the mixed detection solution, donor beads and acceptor beads are both 5μg/mL. Cover the detection board with tin foil, and place it in the dark at room temperature for 1 hour. The signal was read using the AlphaScreen detector on Spectra max i3. Standardize the AlphaScreen signal based on the readings obtained by the positive control (maximum signal control) and negative control (minimum signal control) to give the inhibition rate of different concentrations of the compound, and then use GraphPad Prism5 for nonlinear regression analysis, by Y=Bottom+( Top-Bottom) / (1 + 10 ^ ((LogIC50-X) * HillSlope)) to make a dose response inhibition curve equation, to obtain IC ₅₀ values for each compound.

In order to eliminate the false positives caused by the compound interfering with the AlphaScreen detection system, the compound was diluted in the same way, and the biotinylated peptide Biotinylated-(His) _{6 was used to} replace the EED and peptide H3K27me3 in the detection system. After incubating for the same time, in Spectra Read the signal value on max i3. Process the data in the same way.

result

The following table shows the IC ₅₀ values of some compounds of the present invention.

The letter A represents IC _{50 is} less than 20nM;

The letter B represents an IC ₅₀ of 20 nM to 100 nM.

table 3

Example 55: ELISA (H3K27 trimethylation) analysis

A representative compound of the present disclosure was diluted 3-fold in DMSO, 10 concentration gradients were detected for each compound, and the highest measured concentration was 10 μM. The compound was diluted 200-fold into G401 cells cultured in 96-well plates (final concentration of DMSO was 0.5%). After 72 hours of incubation of the administered cells, the level of histone H3K27 trimethylation was detected by ELISA.

Histone extraction: The cells treated with the compound in a 96-well plate were treated with 1x PBS (10x PBS buffer (80g NaCl (Sigma, product number S3014)), 2g KCl (Sigma, product number 60128), 14.4g Na ₂ HP04 (Sigma, Product number S5136), 2.4g KH ₂ P04 (Sigma, product number P9791) in 1L of water, pH to 7.4) wash three times, add 100μL 0.4N HCl to each well, place at 4°C, gently shake for 2 hours to lyse cells. Neutralize the lysate with 80μL neutralization buffer (0.5M disodium hydrogen phosphate, pH12.5, 2.5mM DTT; 1% cocktail (Sigma, product number P8340)) (fully mix the cell lysate and neutralization buffer) .

ELISA detection method: transfer the cell lysate in parallel to two 384-well detection plates (PerkinElmer, OptiPlate-384HB, product number 6007290), one plate is used to detect the level of H3K27 trimethylation, the other plate is used to determine H3 The final volume of PBS was adjusted to 50μL/well and coated overnight at 4°C. The next day, discard the solution in the well, and use TBST buffer (l xTBS (10x TBS: 24.2g Tris (Sigma, product number T6066), 80g NaCl (Sigma, product number S3014)) into 1L of water, adjust the pH to 7.6 with HCl) , 0.1% Tween-20) wash 5 times, dry the water on absorbent paper. 70 μL of blocking buffer (TBST, 5% BSA) was added to the coated reaction well and incubated at room temperature for 1 hour. Discard the blocking buffer and add the primary antibody (30 μL/well). The required primary antibodies are diluted with blocking buffer, the dilution factor is as follows: anti-H3K27me3 antibody (Cell Signaling Technology, product number 9733), 1:2000 dilution; anti-H3 antibody (Cell Signaling Technology, product number 4499), 1:10000 dilution. After adding the primary antibody, incubate at room temperature for 1 hour. After washing 5 times with TBST, dry the water, add secondary antibody (30μL/well) to each reaction well,

Example 56: Cell Proliferation Analysis

Using standard cell culture conditions, human B cell non-Hodgkin lymphoma cells KARPAS-422S were cultured in culture flasks. The medium is 15% fetal bovine serum (FBS, Invitrogen, product number 10099-141), 1% penicillin/streptomycin solution (P/S) RPMI-1640 (Invitrogen, product number 11875), and the culture flask is placed at a temperature Cultivate in a sterile incubator at 37°C, relative humidity 95%, and 5% CO2. In order to examine the effect of inhibitors on cell proliferation PRC2, taken in exponential growth phase cells, at a density of 1x10 ⁴ cells / well were seeded into 96-well plates (Corning, product number 3904), 100μL medium was added to each well. Subsequently, different concentrations of the compounds disclosed herein were added to the wells of the inoculated cells (9 concentration gradients were set for each compound, the highest detection concentration was 10 μM, 3-fold gradient dilution), 2 for each treatment concentration Repeat in parallel, with a final DMSO concentration of 0.5%. After that, the number of viable cells was measured with Vi-CELL (Beckman Coulter) every 3 to 4 days. The counted cells each time were seeded into a new 96-well plate at the same density ( ¹ ×10 ⁴ cells/well), supplemented with fresh medium to 100 μL, and added compounds of different concentrations at the same time. Incubate to the 13th day, add 100 μL of CellTiter-Glo (CellTiter-GloCellTiter-GloCellTiter-GloCTG) (Promega, product number G7573) to each well, and place in the dark at room temperature for 10-20 minutes. Use Molecular Devices, SpectraMax i3X to read the luminescence signal. GraphPad prisim5 was used to fit the data to a dose response curve to obtain the IC ₅₀ value of the test compound.

result

The following table shows the IC ₅₀ values of some compounds of the present invention. The letter A represents IC _{50 is} less than 20nM;

The letter B represents an IC ₅₀ of 20nM to 100nM;

The letter C represents an IC ₅₀ of 100nM or more.

table 5

The compounds disclosed in the present invention can be used to treat cancers related to the mechanism of action of the EED protein and/or PRC2 protein complex, including but not limited to lymphoma, leukemia, and multiple diseases such as diffuse large B-cell lymphoma, follicular lymphoma, etc. Myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, liver cancer, prostate cancer, breast cancer, brain tumors including neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon Cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, kidney cancer, rectal cancer, thyroid cancer, parathyroid tumor, uterine tumor and soft tissue sarcoma, etc.

Claims (17)

A compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate or its isotope-labeled compound,

among them, X is independently C or N; R ^{2 is} independently H or halogen; R ^{3 is} independently H or halogen; R ^{4 is} independently H or halogen; n is independently 0 or 1; When X=C, then n is 1, forming a pyridopyrimidine structure

When X=N, then n is 0, forming a triazolopyrimidine structure

R ^{1 is} independently hydrogen, halogen, cyano, R ^1a substituted or unsubstituted C _1-8 alkyl, C _1-8 haloalkyl, R ^1a substituted or unsubstituted C _3-8 cycloalkyl, R ^1b Substituted or unsubstituted C _3-8 heteroalkyl, R ^1c substituted or unsubstituted alkenyl, R ^1b substituted or unsubstituted C _5-8 cycloalkenyl, R ^1b substituted or unsubstituted C _5-8 hetero Cycloalkenyl, 0-3 R ^1d substituted C _6-10 aryl or 0-3 R ^1d substituted "having C _1-20 carbon atoms and 1-4 independently selected from N, NR ^1de1 , O or S(O) _0-2 heteroatom heteroaryl"; Each of R ^1a and R ^{1b is} independently C _1-4 alkoxy, -C(=O)NR ^1de1 R ^1de2 , C _1-4 alkyl, C _1-4 haloalkyl, amino, protecting group protection The amino, halogen, hydroxyl or -C(=O)OR ^1de4 ; Each R ^{1c is} independently a C _1-4 alkyl or ester group; R ^{1d is} independently halogen, R ^1a substituted or unsubstituted C _1-4 alkoxy, R ^1a substituted or unsubstituted C _1-4 alkyl, -C(=O)NR ^1de1 R ^1de2 , -S( =0) ₂ R ^1de3 , NR ^1de1 R ^1de2 , R ^5c substituted or unsubstituted C _3-10 cycloalkyl, R ^5c substituted or unsubstituted "having C _1-20 carbon atoms and 1-4 independently selected from NR ^1de1 , N, O or S(O) _0-2 heteroatom heterocyclic group"; Each R ^{5c is} independently halogen, C _1-4 alkyl, C _1-4 alkoxy, -C(=O)NR ^1de1 R ^1de2 or C _1-4 haloalkyl; Each R ^{1de1 is} independently hydrogen or C _1-4 alkyl; Each R ^{1de2 is} independently hydrogen or C _1-4 alkyl; Each of R ^1de3 and R ^1de4 is a C _1-4 alkyl group. The compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate, or its isotope-labeled compound according to claim 1, wherein: In R ² , the halogen is fluorine; And/or, in R ³ , the halogen is fluorine; And/or, in R ⁴ , the halogen is fluorine; And/or, in R ¹ , the halogen is fluorine, bromine or iodine; And / or, wherein R ^1, R ^1a according to a substituted or unsubstituted C _1-8 alkyl in C _1-8 alkyl is C _1-4 alkyl; preferably methyl, ethyl, n-propyl Or isopropyl; And/or, in R ¹ , the C _1-8 haloalkyl group is a C _1-4 haloalkyl group; And / or, R ^1, R ^1a according to a substituted or unsubstituted C _3-8 cycloalkyl C _3-8 cycloalkyl group is a cyclohexyl group; C _3-8 heteroalkyl, and / or, wherein R ^1, R ^1b according to a substituted or unsubstituted alkyl C _3-8 heteroalkyl having from 1 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur Atom C _3-8 heteroalkyl; preferably

And/or, in R ¹ , the alkenyl group in the substituted or unsubstituted alkenyl group of R ^1c is a C _2-10 alkenyl group; preferably a C _2-6 alkenyl group, more preferably a vinyl group; And/or, in R ¹ , the substituted or unsubstituted C _5-8 cycloalkenyl group of R ^1b is

And / or, wherein R ^1, R ^1b according to a substituted or unsubstituted heterocyclyl C _5-8 alkenyl group having 1 to 3 substituents independently selected from nitrogen, C _5-8 heterocyclic ring oxygen, or sulfur Alkenyl, more preferably a C _5-8 heterocycloalkenyl having one nitrogen, oxygen or sulfur heteroatom; for example

And/or, in R ¹ , the C _6-10 aryl group in the 0-3 R ^1d- substituted C _6-10 aryl group is a phenyl group; And/or, in R ¹ , the 0-3 R ^1d substituted "having C _1-20 carbon atoms and 1-4 independently selected from N, NR ^1de1 , O or S(O) _0-2 The heteroaryl group in the "heteroatomic heteroaryl group" has C _1-10 carbon atoms; the heteroaryl group is preferably a 5-12 membered heteroaryl group; more preferably has C _3-5 carbon atoms and 1-2 Heteroaryl groups independently selected from heteroatoms of N and O; for example, furyl, pyridyl, pyrazolyl, isothiazolyl or pyrimidinyl, for example

And/or, in R ^1a and R ^1b , the C _1-4 alkyl group is methyl; And/or, in R ^1a and R ^1b , the C _1-4 haloalkyl group is trifluoromethyl; And/or, in R ^1a and R ^1b , the protecting group in the amino group protected by the protecting group is

R ^5a is a C _{1- 4} alkyl or C _1-4 alkoxy; preferably

Can be tert-butoxycarbonyl; And/or, in R ^1a and R ^1b , the halogen is fluorine; And/or, in R ^1a and R ^1b , -C(=O)OR ^1de4 is -C(=O)OC(CH ₃ ) ₃ ; And/or, in R ^1c , the ester group is

Wherein R ^5b is C _1-4 alkyl; And/or, in R ^1d , the halogen is fluorine, chlorine, bromine or iodine; And / or, R ^1d, the R ^1a is a substituted or unsubstituted C _1-4 alkoxy C _1-4 alkoxy group is methoxy; And / or, R ^1d, the R ^1a is a substituted or unsubstituted C _1-4 alkyl in C _1-4 alkyl is methyl, ethyl, n-propyl or isopropyl; and / or , In R ^1d , the -C(=O)NR ^1de1 R ^1de2 is -C(=O)NH ₂ ; And/or, in R ^1d , the -S(=O) ₂ R ^1de3 is -S(=O) ₂ Me; And/or, in R ^1d , the NR ^1de1 R ^1de2 is -NH ₂ ; And/or, in R ^1d , the substituted or unsubstituted C _3-10 cycloalkyl group of R ^5c is a C _3-8 cycloalkyl group; And/or, in R ^1d , R ^5c substituted or unsubstituted "heteroatoms having C _1-20 carbon atoms and 1-4 heteroatoms independently selected from NR ^1de1 , N, O or S(O) _0-2 The heterocyclic group in "cyclic group" has C _1-10 carbon atoms; the heterocyclic group is preferably 5-12 membered heterocyclic group, more preferably C _3-5 carbon atoms and 1-2 selected from N and O heterocyclic groups. Heterocyclic group of atoms; for example

The compound of formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate, or its isotope-labeled compound as claimed in claim 1, wherein R ¹ is hydrogen, halogen, cyano, C _{1- 8} alkyl, C _1-8 haloalkyl, or one of the following structures:

among them, Each j is independently 0, 1, 2 or 3; k is 0, 1, 2, 3 or 4; each V is independently C, N or O, and there are at most two N or at the same time in the same ring O; Ring A is a substituted or unsubstituted C _5-10 heteroaryl group containing 1-3 heteroatoms; each ring B is independently a substituted or unsubstituted C _5-10 containing 1-4 heteroatoms Heteroaryl, wherein the heteroatom is independently N, O, or S. The compound of formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate, or its isotope label according to any one of claims 1 to 3 Compound, where: R ¹ is R ^1b substituted or unsubstituted C _5-8 heterocycloalkenyl, 0-3 R ^1d substituted C _6-10 aryl or 0-3 R ^1d substituted with C _1-20 carbon atoms And 1-4 heteroaryl groups independently selected from heteroatoms of N, NR ^1de1 , O or S(O) _0-2 ; Preferably, R ^1b is -C(=O)OR ^1de4 ; Preferably, the number of R ^1d is 0, 1, or 2; Preferably, each R ^{1d is} independently halogen, R ^1a substituted or unsubstituted C _1-4 alkoxy, R ^1a substituted or unsubstituted C _1-4 alkyl, -C(=O)NR ^1de1 R _{^{1de2, -S (= O) 2}} R 1de3, -NR 1de1 R 1de2 R ^5c, or a substituted or unsubstituted C _{1- 20} carbon atoms and 1-4 groups independently selected from ^NR 1de1, N, O or S (O ) _0-2 heteroatom heterocyclic group; Preferably, each R ^{1a is} independently halogen or hydroxyl; Preferably, R ^1de1 and R ^1de2 are hydrogen. The compound of formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate, or its isotope-labeled compound as claimed in claim 4, wherein: R ^1b R ¹ is a substituted or unsubstituted heterocyclyl C _5-8 alkenyl group; R ^1b is ^-C (= O) OR 1de4; Alternatively, R ¹ is 0-3 R ^1d substituted C _6-10 aryl groups, each R ^{1d is} independently halogen, R ^1a substituted or unsubstituted C _1-4 alkoxy, R ^1a substituted or unsubstituted The C _1-4 alkyl group, -C(=O)NR ^1de1 R ^1de2 , -S(=O) ₂ R ^1de3 or NR ^1de1 R ^1de2 ; each R ^{1a is} independently halogen; R ^1de1 and R ^1de2 are hydrogen ； Alternatively, R ¹ is 0-3 R ^1d substituted "heteroaromatics having C _1-20 carbon atoms and 1-4 heteroatoms independently selected from N, NR ^1de1 , O or S(O) _0-2 Group; each R ^{1d is} independently halogen, R ^1a substituted or unsubstituted C _1-4 alkoxy, R ^1a substituted or unsubstituted C _1-4 alkyl, NR ^1de1 R ^1de2 or R ^5c substituted or unsubstituted Substituted heterocyclic group with C _1-20 carbon atoms and 1-4 heteroatoms independently selected from NR ^1de1 , N, O or S(O) _0-2 ; each R ^{1a is} independently halogen or hydroxyl; R ^1de1 and R ^1de2 are hydrogen. The compound of formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate, or its isotope label according to any one of claims 1 to 3 Compound, where R ¹ is fluorine, bromine, iodine,

And/or, in the compound represented by formula (I)

for

The compound of formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate, or its isotope label according to any one of claims 1-6 A compound, the compound represented by formula (I) is a compound represented by the following formula (Ia) or (Ib),

Wherein, R ¹ , R ² , R ³ and R ⁴ are as defined in any one of claims 1-6. The compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate, or its isotope-labeled compound according to claim 7, wherein: In the compound represented by formula (Ia), R ¹ is substituted with 0-3 R ^1d "having C _1-20 carbon atoms and 1-4 independently selected from N, NR ^1de1 , O or S(O) _{0- 2} heteroatom heteroaryl groups; Preferably, the number of R ^1d is 0, 1, or 2; Preferably, each R ^{1d is} independently halogen, R ^1a substituted or unsubstituted C _1-4 alkoxy (such as methoxy), R ^1a substituted or unsubstituted C _1-4 alkyl, NR ^1de1 R ^1de2 or a heterocyclic group having C _1-20 carbon atoms and 1-4 heteroatoms independently selected from NR ^1de1 , N, O or S(O) _0-2 ; Preferably, each R ^{1a is} independently halogen and hydroxyl; Preferably, R ^1de1 and R ^1de2 are hydrogen; Or, in the compound represented by formula (Ib), R ¹ is 0-3 R ^1d substituted "having C _1-20 carbon atoms and 1-4 independently selected from N, NR ^1de1 , O or S(O) Heteroaryl with a heteroatom of _0-2 "; Preferably, the number of R ^1d is 0, 1, or 2; Preferably, each R ^{1d is} independently halogen, C _1-4 alkyl, -C(=O)NR ^1de1 R ^1de2 or -S(=O) ₂ R ^1de3 . The compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate or its isotope-labeled compound according to claim 7 or 8, wherein , In the compound represented by formula (Ia): R ¹ is

Or, in the compound represented by formula (Ib): R ¹ is

The compound of formula (I), its pharmaceutically acceptable salt, hydrate, prodrug, stereoisomer, its solvate or its isotope-labeled compound according to any one of claims 1-9, wherein The compound is selected from any of the following compounds:

A method for preparing the compound represented by formula (I) according to any one of claims 1-10, which comprises the following steps: The halogenated intermediate compound A undergoes coupling reaction to obtain the compound of formula (I),

Wherein, W represents halogen, preferably Br; X, n, R ¹ , R ² , R ³ and R ⁴ are as defined in any one of claims 1-10; The method for preparing the compound represented by formula (I) preferably includes the following steps: the halogenated intermediate A and the compound R ¹ -W ¹ undergo the Suzuki reaction shown below;

Wherein, W represents halogen, preferably Br; X, n, R ¹ , R ² , R ³ and R ⁴ are as defined in any one of claims 1-10, and W ¹ is boric acid, borate or borate (E.g. -B(OH) ₂ or

). A preparation method of halogenated intermediate compound A, which comprises the following steps: Chloropyridine, pyrimidine or pyrazine intermediate B is substituted with intermediate C under basic conditions to obtain halogenated intermediate compound A,

Wherein, W, X, n, R ² , R ³ and R ⁴ are as defined in claim 11. A halogenated intermediate compound A,

The definitions of W, X, n, R ² , R ³ and R ⁴ are as described in claim 11. The compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate, or its isotope according to any one of claims 1-10 The application of the labeled compound in the preparation of a drug, the drug is preferably an EED inhibitor or an EZH2 inhibitor or a PRC2 inhibitor, the drug can be used to treat cancer, and the cancer preferably interacts with EED and/or PRC2 protein complex Mechanism related. The use according to claim 14, wherein the cancer is selected from lymphomas such as diffuse large B-cell lymphoma, follicular lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, liver cancer, Prostate cancer, breast cancer, brain tumors including neuroblastoma, glioma, glioblastoma and astrocytoma, cervical cancer, colon cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer , Nasopharyngeal cancer, ovarian cancer, pancreatic cancer, kidney cancer, rectal cancer, thyroid cancer, parathyroid tumor, uterine tumor and soft tissue sarcoma; preferably spreading large B-cell lymphoma or follicular lymphoma; Preferably, the compound represented by formula (I), its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, its solvate or its isotope-labeled compound are used in combination with other drugs ; Preferably, the other drugs are selected from anti-cancer drugs, tumor immune drugs, anti-allergic drugs, antiemetics, analgesics or cytoprotective drugs. A pharmaceutical composition comprising the compound represented by formula (I) according to any one of claims 1-10, its pharmaceutically acceptable salt, its hydrate, its prodrug, and its stereoisomers Body, its solvate or its isotope-labeled compound, and pharmaceutically acceptable excipients. A pharmaceutical preparation, comprising the compound represented by formula (I) according to any one of claims 1-10, its pharmaceutically acceptable salt, its hydrate, its prodrug, its stereoisomer, The solvate or isotope-labeled compound thereof, and the pharmaceutical preparation is selected from the group consisting of tablets, capsules, pills, powders, granules, elixirs, tinctures, suspensions, syrups, emulsions and solutions; Preferably, the administration mode of the pharmaceutical preparation is selected from oral, sublingual, subcutaneous, intravenous, intramuscular, intrasternal injection, nasal administration, topical administration and rectal administration; and/or, so The pharmaceutical preparation is taken single or multiple times a day.