WO2025051157A1 - Pyridine-2(1h)-ketone prmt5-mta inhibitor, and pharmaceutical composition and use thereof - Google Patents

Abstract

Provided is a type of compound acting as a PRMT5-MTA inhibitor, specifically a compound represented by formula (I), a stereoisomer, a deuterate, a solvate, and a pharmaceutically acceptable salt thereof, the symbols being as defined in the text. Further provided are a pharmaceutical composition comprising the compound, and a use thereof in treating cancer.

Description

A pyridin-2(1H)-one PRMT5-MTA inhibitor, its pharmaceutical composition and its application Technical field:

The present invention belongs to the field of medicine, and specifically relates to a class of pyridine-2(1H)-one compounds, their stereoisomers, deuterated products, solvates, prodrugs, metabolites, pharmaceutically acceptable salts, pharmaceutical compositions containing the same, and uses thereof in the preparation of drugs for treating PRMT5-mediated related diseases.

Background technology:

Protein arginine methyltransferase (PRMT) can methylate histones and non-histones, thus participating in the regulation of biological processes such as gene transcription, signal transduction, protein stability, cell proliferation, differentiation, apoptosis and tumor formation. Currently, 11 PRMT family members have been discovered, which can be divided into types I, II and III according to the different ways of catalyzing arginine methylation. Among them, PRMT5 belongs to type II, and its catalytic form is symmetrical dimethylation.

PRMT5 catalyzes the transfer of the methyl group on S-adenosylmethionine (SAM) to the guanidine N atom on the substrate arginine, causing symmetric dimethylation (SDMA). It plays an important role in regulating various cellular processes, such as transcription, RNA splicing, translation, and DNA damage response. PRMT5 protein is overexpressed in many types of cancer, including B and T cell lymphoma, metastatic melanoma, neuroblastoma, glioblastoma, ovarian cancer, breast cancer, etc., and increasing evidence shows that it plays an important role in tumor occurrence and development. On this basis, PRMT5 inhibitors have become a hot spot for the research and development of tumor treatment drugs.

CDKN2A is a tumor suppressor gene located on human chromosome 9p21. The loss of CDKN2A occurs in about 10-15% of all human cancers, including 53% of glioblastomas, 26% of pancreatic cancers, and other tumor types (K.J.Mavrakis et al., Science 10.1126/science.aad5944 (2016)). The metabolic enzyme 5-methylthioadenosine phosphorylase (MTAP) gene is also located on chromosome chr9p21. Because it is very close to the location of CDKN2A, MTAP loss is often accompanied in tumors with CDKN2A gene loss. Therefore, MTAP is also one of the genes with the highest mutation frequency in tumors.

In 2016, a paper published in Science revealed for the first time that MTAP deficiency and PRMT5 have synthetic lethal effects (K.J.Mavrakis et al., Science 10.1126/science.aad5944 (2016)). MTAP deficiency causes its substrate methylthioadenosine (MTA) to accumulate. MTA is an analog of SAM and can selectively inhibit the activity of PRMT5. MTA competes with SAM and binds to PRMT5 to form a PRMT5-MTA complex. By strengthening the inhibition of the PRMT5-MTA complex, PRMT5 activity can be specifically inhibited in tumor cells, while the inhibitory effect on PRMT5 activity in normal cells is weak.

A large number of PRMT5 small molecule inhibitors have been reported. Depending on whether the compounds occupy the SAM binding site, they can be divided into two categories: SAM non-competitive inhibitors and SAM competitive inhibitors. SAM non-competitive inhibitors occupy the substrate binding site and compete with the substrate, while SAM competitive inhibitors occupy the SAM binding site.

Although SAM non-competitive inhibitors can effectively inhibit the in vitro growth of many cell lines, they lack tumor cell line selectivity, and their ability to inhibit tumor cells is independent of the status of MTAP in cells, which may lead to potential clinical toxicity risks. In cancer cells lacking MTAP, PRMT5-MTA inhibitors selectively affect only tumor cells and do not affect healthy cells, thus providing a therapeutic safety window. Therefore, designing a small molecule that cooperates with MTA to bind to PRMT5 and specifically binds to the PRMT5·MTA complex to inhibit tumor cell growth is an effective means to improve the selectivity of PRMT5 inhibitors.

Summary of the invention

The present invention provides a compound represented by formula (I), and its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts:

in,

represents any single or double bond allowed by the valence;

X ₁ and X ₂ are independently selected from NR ^X , S, N, CR ^X1 ;

R ^X and R ^X1 are independently selected from H, halogen, -C ₁ ～C ₆ alkyl, deuterium-substituted -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl, -C(O)-C ₁ ～C ₃ alkyl;

X ₃ is selected from N, CR ³ ;

R ³ is selected from H, halogen, -C ₁ ～C ₆ alkyl, -C(O)NH ₂ , -C(O)NH(C ₁ ～C ₃ alkyl), -C(O)N(C ₁ ～C ₃ alkyl) ₂ ;

X ₄ is selected from N, CR ⁵ ;

R ⁷ is selected from H, halogen, -CN, -NO ₂ , -NH ₂ , -C ₁ ～C ₆ alkyl, deuterium-substituted -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl;

L is selected from a bond,

n is independently selected from 0, 1, and 2;

R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ～C ₆ alkyl, -C ₂ ～C ₆ alkenyl, halogen-substituted -C ₁ ～C ₆ alkyl, -OC ₁ ～C ₆ alkyl, 3-6-membered carbocyclic ring, 5-10-membered heteroaromatic ring, 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring, aromatic ring may be optionally substituted by one or more halogen, -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -OC ₁ ～C ₃ alkyl;

R ⁴ and R ⁶ are independently selected from hydrogen, -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl, -C ₀ ～C ₂ alkylene-(3-10 membered carbocyclic ring), -C ₀ ～C ₂ alkylene-(4-11 membered heterocyclic ring), -C ₀ ～C ₂ alkylene-(6-10 membered aromatic ring) or -C ₀ ～C ₂ alkylene-(5-10 membered heteroaromatic ring); wherein the alkyl, alkylene, carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ⁴¹ ;

R ⁴¹ is selected from hydrogen, halogen, amino, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₆ alkyl, -C ₂ ～C ₆ alkenyl, -C ₂ ～C ₆ alkynyl, halogen-substituted -C ₁ ～C ₆ alkyl, halogen-substituted -C ₂ ～C ₆ alkenyl, halogen-substituted -C ₂ ～C ₆ alkynyl, -OC ₁ ～C ₆ alkyl;

R ² is selected from a 3-10-membered carbocyclic ring, a 4-11-membered heterocyclic ring, a 6-10-membered aromatic ring or a 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ²¹ ;

^R21 is selected from hydrogen, halogen, nitro, cyano, oxo, _-NH2 , -C(O) _NH2 , -C(O)H, -C(O)OH, _-C1 ~ _C6 alkyl, _-C2 ~ _C6 alkenyl, _-C2 ~ _C6 alkynyl, halogen-substituted _-C1 ~ _C6 alkyl, halogen-substituted _-C2 ~ _C6 alkenyl, halogen-substituted _-C2 ~ _C6 alkynyl, _-OC1 ~ _C6 alkyl, -OC2~C6 _alkenyl , _-OC2 ~C6 alkynyl, -O _- halogen-substituted _C1 ~ _C6 alkyl, -O-halogen-substituted _C2 ~ _{C6 alkenyl} , -O-halogen-substituted _C2 ~C6 _6- alkynyl, -O-(3-6-membered carbocyclic ring), 3-10-membered carbocyclic ring, 4-11-membered heterocyclic ring, 6-10-membered aromatic ring or 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ^c ;

R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ~ C ₆ alkyl, -C ₂ ~ C ₆ alkenyl, -C ₂ ~ C ₆ alkynyl, halogen-substituted -C ₁ ~ C ₆ alkyl, halogen-substituted -C ₂ ~ C ₆ alkenyl, halogen-substituted -C ₂ ~ C ₆ alkynyl, -OC ₁ ~ C ₆ alkyl, -OC ₂ ~ C ₆ alkenyl, -OC ₂ ~ C ₆ alkynyl, -O-halogen-substituted C ₁ ~ C ₆ alkyl, -O-halogen-substituted C ₂ ~ C ₆ alkenyl or -O-halogen-substituted C ₂ ~ C ₆ alkynyl.

In certain specific embodiments, the compound of formula (I), its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts are as shown in formula (IIa) or formula (IIb):

wherein R ¹ , R ² , R ⁷ , ^RX , ^RX1 , L, X ₃ and X ₄ are as described in formula (I).

In certain specific embodiments, the compound of formula (I), its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts are as shown in formula (IIc), formula (IId), formula (IIe), or formula (IIf):

Wherein, R ¹ , R ² , R ³ , R ⁷ , ^RX , ^RX1 , L and X ₄ are as described in formula (I).

In certain specific embodiments, the compound represented by formula (I), its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts are represented by formula (Ia):

in,

represents any single or double bond allowed by the valence;

R ¹ , R ² , R ⁵ , X ₁ , X ₂ and L are as described in formula (I).

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), a stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt thereof,

Wherein, R ⁷ is selected from H, halogen, -CN, -NO ₂ , -NH ₂ , -C ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl substituted with deuterium, -C ₁ ～C ₃ alkyl substituted with halogen.

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), a stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt thereof,

Wherein, ^R7 is selected from H, F, Cl, Br, I, -CN _{, -NO2} , _-NH2 , methyl, ethyl, _propyl , isopropyl, _-CF3 , _-CH2F , _-CHF2 , _-CH2CH2F , _-CH2CHF2 , _-CH2CF3 , _{-CH2D , -CH2CD3 .}

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), a stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt thereof,

Wherein, R ³ is selected from H, halogen, -C ₁ -C ₃ alkyl, -C(O)NH ₂ , -C(O)NH(C ₁ -C ₃ alkyl), -C(O)N(C ₁ -C ₃ alkyl) ₂ .

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), a stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt thereof,

Wherein, R ³ is selected from H, F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, -C(O)NH ₂ , -C(O)NHCH ₃ , -C(O)N(CH ₃ ) ₂ .

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), a stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt thereof.

Wherein, X ₄ is selected from N, CR ⁵ ;

R ⁵ is independently selected from hydrogen, halogen, -C ₁ ~ C ₃ alkyl, -C ₂ ~ C ₄ alkenyl, halogen-substituted -C ₁ ~ C ₃ alkyl, -OC ₁ ~ C ₃ alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring and aromatic ring may be optionally substituted by one or more halogen, -C ₁ ~ C ₃ alkyl, halogen-substituted -C ₁ ~ C ₃ alkyl and -OC ₁ ~ C ₃ alkyl.

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe), formula (IIf) or formula (Ia), stereoisomers, deuterated forms, solvates, pharmaceutically acceptable salts thereof,

Wherein, R ^X and R ^X1 are independently selected from H, halogen, -C ₁ ～C ₃ alkyl, deuterium-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, and -C(O)CH ₃ .

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe), formula (IIf) or formula (Ia), stereoisomers, deuterated forms, solvates, pharmaceutically acceptable salts thereof,

Wherein, ^RX and ^RX1 are independently selected from H, F, Cl, Br, _{I ,} methyl _, ethyl, _propyl , isopropyl, -CF3, -CH2F, _-CHF2 , _{-CH2CH2F , -CH2CHF2} , -C(O) _CH3 , _-CH2CF3 , _-CD3 , _-CH2D , _{-CH2CD3 .}

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe), formula (IIf) or formula (Ia), stereoisomers, deuterated forms, solvates, pharmaceutically acceptable salts thereof,

Wherein, ^R1 and ^R5 are independently selected from hydrogen, halogen, _-C1 ~ _C3 alkyl, _-C2 ~ _C4 alkenyl, halogen-substituted _-C1 ~ _C3 alkyl, _-OC1 ~ _C3 alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring and aromatic ring may be optionally substituted by one or more halogen, _-C1 ~ _C3 alkyl, halogen-substituted _-C1 ~ _C3 alkyl and _-OC1 ~ _C3 alkyl.

In certain specific embodiments, the compound represented by formula (I), its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts are represented by formula (Ia):

in,

X ₁ and X ₂ are independently selected from NH, S, N, and CH;

represents any single or double bond allowed by the valence;

L is selected from a bond,

n is independently selected from 0, 1, and 2;

^R1 and ^R5 are independently selected from hydrogen, halogen, _-C1 - _C6 alkyl, halogen-substituted _-C1 - _C6 alkyl, _-C2 - _C6 alkenyl- _OC1 - _C6 alkyl, 3-6-membered carbocyclic ring, 5-10-membered heteroaromatic ring, 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring, aromatic ring can be optionally substituted by one or more halogen, -C ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl substituted with halogen, -OC ₁ ～C ₃ alkyl substituted; for example, R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ～C ₆ alkyl, -C ₁ ～C ₆ alkyl substituted with halogen;

R ⁴ and R ⁶ are independently selected from hydrogen, -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl, -C ₀ ～C ₂ alkylene-(3-10 membered carbocyclic ring), -C ₀ ～C ₂ alkylene-(4-11 membered heterocyclic ring), -C ₀ ～C ₂ alkylene-(6-10 membered aromatic ring) or -C ₀ ～C ₂ alkylene-(5-10 membered heteroaromatic ring); wherein the alkyl, alkylene, carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ⁴¹ ;

R ⁴¹ is selected from hydrogen, halogen, amino, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₆ alkyl, -C ₂ ～C ₆ alkenyl, -C ₂ ～C ₆ alkynyl, halogen-substituted -C ₁ ～C ₆ alkyl, halogen-substituted -C ₂ ～C ₆ alkenyl, halogen-substituted -C ₂ ～C ₆ alkynyl, -OC ₁ ～C ₆ alkyl;

R ² is selected from a 3-10-membered carbocyclic ring, a 4-11-membered heterocyclic ring, a 6-10-membered aromatic ring or a 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ²¹ ;

^R21 is selected from hydrogen, halogen, nitro, cyano, oxo, _-NH2 , -C(O) _NH2 , -C(O)H, -C(O)OH, _-C1 ~ _C6 alkyl, _-C2 ~ _C6 alkenyl, _-C2 ~ _C6 alkynyl, halogen-substituted _-C1 ~ _C6 alkyl, halogen-substituted _-C2 ~ _C6 alkenyl, halogen-substituted _-C2 ~ _C6 alkynyl, _-OC1 ~ _C6 alkyl, -OC2~C6 _alkenyl , _-OC2 ~C6 alkynyl, -O _- halogen-substituted _C1 ~ _C6 alkyl, -O-halogen-substituted _C2 ~ _{C6 alkenyl} , -O-halogen-substituted _C2 ~C6 _6- alkynyl, -O-(3-6-membered carbocyclic ring), 3-10-membered carbocyclic ring, 4-11-membered heterocyclic ring, 6-10-membered aromatic ring or 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ^c ;

R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ~ C ₆ alkyl, -C ₂ ~ C ₆ alkenyl, -C ₂ ~ C ₆ alkynyl, halogen-substituted -C ₁ ~ C ₆ alkyl, halogen-substituted -C ₂ ~ C ₆ alkenyl, halogen-substituted -C ₂ ~ C ₆ alkynyl, -OC ₁ ~ C ₆ alkyl, -OC ₂ ~ C ₆ alkenyl, -OC ₂ ~ C ₆ alkynyl, -O-halogen-substituted C ₁ ~ C ₆ alkyl, -O-halogen-substituted C ₂ ~ C ₆ alkenyl or -O-halogen-substituted C ₂ ~ C ₆ alkynyl.

In certain specific embodiments, the compound of formula (Ia), its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts are as shown in formula (Ie):

wherein R ¹ , R ² , R ⁵ and L are as described in formula (Ia).

In certain specific embodiments, the compound of formula (Ia), its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts are as shown in formula (Ib), formula (Ic), or formula (Id):

Wherein, R ¹ , L, and R ² are as described in formula (Ia).

In certain specific embodiments, the present invention relates to a compound of Formula (I), Formula (Ia), Formula (Ib), Formula (Ic), Formula (Id), Formula (Ie), Formula (IIa), Formula (IIb), Formula (IIc), Formula (IId), Formula (IIe), or Formula (IIf), a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, wherein:

R ² is selected from a 3-7-membered carbocyclic ring, a 4-6-membered monocyclic heterocyclic ring, a 9-11-membered fused heterocyclic ring, a 6-10-membered aromatic ring, or a 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ²¹ ;

^R21 is selected from hydrogen, halogen, nitro, cyano, oxo, _-NH2 , -C(O) _NH2 , -C(O)H, -C(O)OH, _-C1 ~ _C3 alkyl, _-C2 ~ _C3 alkenyl, _-C2 ~ _C3 alkynyl, halogen-substituted _-C1 ~ _C3 alkyl, halogen-substituted _-C2 ~ _C3 alkenyl, halogen-substituted _-C2 ~ _C3 alkynyl, _-OC1 ~ _C3 alkyl, -OC2~C3 _alkenyl , _-OC2 ~C3 alkynyl, -O _- halogen-substituted _C1 ~ _C3 alkyl, -O-halogen-substituted _C2 ~ _{C3 alkenyl} , -O-halogen-substituted _C2 ~C3 _3- alkynyl, -O-(3-6-membered carbocyclic ring), 3-10-membered carbocyclic ring, 4-11-membered heterocyclic ring, 6-10-membered aromatic ring or 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ^c ;

R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, -C ₂ ～C ₄ alkenyl, -C ₂ ～C ₄ alkynyl, halogen-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₂ ～C ₄ alkenyl, halogen-substituted -C ₂ ～C ₄ alkynyl, and -OC ₁ ～C ₃ alkyl.

In certain specific embodiments, the present invention relates to compounds of formula (I), formula (Ia), formula (Ib), formula (Ic), formula (Id), formula (Ie), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), stereoisomers, deuterated forms, solvates, and pharmaceutically acceptable salts thereof, wherein ^R is selected from For example, selected from

wherein R ²¹ is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -OC ₁ ～C ₃ alkyl, -O-halogen-substituted C ₁ ～C ₃ alkyl, -O-(3-6-membered carbocyclic ring), 3-10-membered carbocyclic ring, 4-11-membered heterocyclic ring, 6-10-membered aromatic ring or 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ^c ;

y = 0, 1, 2 or 3;

R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, -C ₂ ～C ₄ alkenyl, -C ₂ ～C ₄ alkynyl, halogen-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₂ ～C ₄ alkenyl, halogen-substituted -C ₂ ～C ₄ alkynyl, and -OC ₁ ～C ₃ alkyl.

In certain specific embodiments, the present invention relates to a compound of Formula (I), Formula (Ia), Formula (Ib), Formula (Ic), Formula (Id), Formula (Ie), Formula (IIa), Formula (IIb), Formula (IIc), Formula (IId), Formula (IIe), or Formula (IIf), a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, wherein:

R ²¹ is selected from hydrogen, halogen, -OC ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl, -O-halogen substituted C ₁ ～C ₃ alkyl, For example, selected from hydrogen, halogen, -OC ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl,

In certain specific embodiments, the present invention relates to compounds of Formula (I), (Ia), (Ib), (Ic), (Id), (Ie), (IIa), (IIb), (IIc), (IId), (IIe) or (IIf), stereoisomers, deuterated forms, solvates, and pharmaceutically acceptable salts thereof, wherein R ^c is selected from hydrogen, F, Cl, Br, nitro, cyano, oxo, -NH ₂ , methyl, ethyl, and isopropyl.

In certain specific embodiments, the present invention relates to a compound of Formula (I), Formula (Ia), Formula (Ib), Formula (Ic), Formula (Id), Formula (Ie), Formula (IIa), Formula (IIb), Formula (IIc), Formula (IId), Formula (IIe) or Formula (IIf), its stereoisomers, deuterated forms, solvates, pharmaceutically acceptable salts, in,

L is selected from a bond,

R ⁴ and R ⁶ are independently selected from hydrogen, -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -C ₀ ～C ₁ alkylene-(3-6 membered carbocyclic ring), -C ₀ ～C ₁ alkylene-(4-6 membered heterocyclic ring), -C ₀ ～C ₁ alkylene-(6-10 membered aromatic ring), -C ₀ ～C ₁ -(5-10 membered heteroaromatic ring); wherein the alkyl, alkylene, carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ⁴¹ ;

R ⁴¹ is selected from hydrogen, halogen, amino, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ -C ₃ alkyl, -C ₂ -C ₃ alkenyl, -C ₂ -C ₃ alkynyl, halogen-substituted -C ₁ -C ₃ alkyl, halogen-substituted -C ₂ -C ₆ alkenyl, halogen-substituted -C ₂ -C ₆ alkynyl, and -OC ₁ -C ₆ alkyl.

In certain specific embodiments, the present invention relates to a compound of formula (I), (Ia), (Ib), (Ic), (Id), (Ie), (IIa), (IIb), (IIc), (IId), (IIe) or (IIf), a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, wherein L is selected from a bond, The a end is connected to ^R2 .

In certain specific embodiments, the present invention relates to compounds of formula (I), (Ia), (Ib), (Ic), (Id), (Ie), (IIa), (IIb), (IIc), (IId), (IIe) or (IIf), stereoisomers, deuterated forms, solvates, and pharmaceutically acceptable salts thereof, wherein ^R4 and ^R6 are independently selected from hydrogen, _-C1 - _C3 alkyl, _-C1 - _C3 alkyl substituted with halogen, 3-6 membered carbocyclic ring, _-CH2- (3-6 membered carbocyclic ring), 4-6 membered heterocyclic ring, _-CH2- (4-6 membered heterocyclic ring), 6-10 membered aromatic ring, _-CH2- (6-10 membered aromatic ring), 5-10 membered heteroaromatic ring, _-CH2 -(5-10 membered heteroaromatic ring); wherein the alkyl, carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ⁴¹ ;

R ⁴¹ is selected from hydrogen, methyl, ethyl, isopropyl.

In certain specific embodiments, the present invention relates to compounds of formula (I), (Ia), (Ib), (Ic), (Id), (Ie), (IIa), (IIb), (IIc), (IId), (IIe) or (IIf), and stereoisomers, deuterated forms, solvates, and pharmaceutically acceptable salts thereof, wherein R ⁴ and R ⁶ are independently selected from a benzene ring, a -CH ₂ -benzene ring, a 5-6 membered heteroaromatic ring, or -CH ₂ -(5-6 membered heteroaromatic ring); wherein the benzene ring and the heteroaromatic ring may be optionally substituted by one or more R ⁴¹ .

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (Ia), formula (Ib), formula (Ic), formula (Id), formula (Ie), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), or a stereoisomer, deuterated product, solvate, or pharmaceutically acceptable salt thereof, wherein R ⁴ and R ⁶ are independently selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl,

In certain specific embodiments, the present invention relates to a compound of formula (I), (Ia), (Ib), (Ic), (Id), (Ie), (IIa), (IIb), (IIc), (IId), (IIe) or (IIf), a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof, wherein L is selected from a bond,

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (Ia), formula (Ie), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), a stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt thereof,

Wherein, R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ~ C ₃ alkyl, halogen-substituted -C ₁ ~ C ₃ alkyl, -C ₂ ~ C ₄ alkenyl, halogen-substituted -C ₁ ~ C ₃ alkyl, -OC ₁ ~ C ₃ alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring, aromatic ring may be optionally substituted by one or more halogen, -C ₁ ~ C ₃ alkyl, halogen-substituted -C ₁ ~ C ₃ alkyl, -OC ₁ ~ C ₃ alkyl; for example, R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ~ C ₃ alkyl or halogen-substituted -C ₁ ~ C ₃ alkyl.

In certain specific embodiments, the present invention relates to compounds of formula (Ib), formula (Ic) or formula (Id), stereoisomers, deuterated forms, solvates, pharmaceutically acceptable salts thereof,

Wherein, R ¹ is selected from hydrogen, halogen, -C ₁ ～C ₃ alkyl or halogen-substituted -C ₁ ～C ₃ alkyl.

In certain specific embodiments, the present invention relates to compounds of formula (Ib), formula (Ic) or formula (Id), stereoisomers, deuterated forms, solvates, pharmaceutically acceptable salts thereof,

wherein R ¹ is selected from hydrogen, methyl, ethyl, Cl, Br, I, F, —CF ₃ , —CH ₂ F, —CHF ₂ , —CH ₂ CH ₂ F, —CH ₂ CHF ₂ or —CH ₂ CF ₃ .

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (Ia), formula (Ie), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), a stereoisomer, deuterated form, solvate, or pharmaceutically acceptable salt thereof,

wherein R ¹ and R ⁵ are independently selected from hydrogen, methyl, ethyl, F, Cl, Br, I, -CF ₃ , -CH ₂ F, -CHF ₂ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ , -CH ₂ CF ₃ , -Cl, -Br, -OCH ₃ , Phenyl, For example, R ¹ and R ⁵ are each independently selected from hydrogen, methyl, ethyl, F, -CF ₃ , -CH ₂ F, -CHF ₂ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ or -CH ₂ CF ₃ .

In certain specific embodiments, the present invention relates to a compound of formula (I), formula (Ia), formula (Ib), formula (Ic), formula (Id), formula (Ie), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), a stereoisomer, deuterated substance, solvate, or pharmaceutically acceptable salt thereof, wherein R ²¹ is selected from H, F, Cl, Br, -CF ₃ , -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl,

In certain specific embodiments, the present invention relates to compounds of formula (I), formula (Ia), formula (Ib), formula (Ic), formula (Id), formula (Ie), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), stereoisomers, deuterated forms, solvates, and pharmaceutically acceptable salts thereof, wherein ^R is selected from For example, selected from

Further, in certain specific embodiments, the present invention relates to compounds of formula (Ib), formula (Ic), formula (Id), and their stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts.

wherein R ¹ is selected from hydrogen, halogen, -C ₁ ～C ₃ alkyl or halogen-substituted -C ₁ ～C ₃ alkyl;

L is selected from (For example, wherein the a end is connected to R ² );

R ⁴ is selected from hydrogen, -C ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl substituted with halogen, 3-6 membered carbocyclic ring or 4-6 membered heterocyclic ring;

R ² is selected from a 3-7 membered carbocyclic ring, a 4-10 membered heterocyclic ring, a 6-10 membered aromatic ring or a 5-10 membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ²¹ .

Further, in certain specific embodiments, the present invention relates to compounds described by formula (Ib), formula (Ic), formula (Id), stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts thereof, wherein R ¹ is selected from hydrogen, methyl, ethyl, F, Cl, Br, I, -CF ₃ , -CH ₂ F, -CHF ₂ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ , or -CH ₂ CF ₃ ;

L is selected from (For example, wherein the a end is connected to R ² );

^R4 is selected from methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl or cyclopentyl;

R ² is selected from in particular

In certain specific embodiments, the present invention relates to compounds of Formula (I), Formula (Ia), Formula (Ie), Formula (IIa), Formula (IIb), Formula (IIc), Formula (IId), Formula (IIe) or Formula (IIf), stereoisomers, deuterated forms, solvates, and pharmaceutically acceptable salts thereof.

in,

R ² is selected from

wherein R ²¹ is selected from hydrogen, halogen, -OC ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl, -O-halogen substituted C ₁ ～C ₃ alkyl,

y = 0, 1, 2 or 3;

wherein R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, -C ₂ ～C ₄ alkenyl, -C ₂ ～C ₄ alkynyl, halogen-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₂ ～C ₄ alkenyl, halogen-substituted -C ₂ ～C ₄ alkynyl, -OC ₁ ～C ₃ alkyl (for example, R ² is specifically selected from );

L is selected from a bond, (For example, L is selected from a bond, Wherein the a end is connected to R ² ),

Wherein, R ⁴ and R ⁶ are independently selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl,

Wherein, R ⁴¹ is selected from hydrogen, halogen, methyl, ethyl, isopropyl (for example, L is specifically selected from a bond, For example, L is specifically selected from a bond, wherein the a end is connected to R ² );

R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ to C ₃ alkyl, -C ₂ to C ₄ alkenyl, halogen-substituted -C ₁ to C ₃ alkyl, -OC ₁ to C ₃ alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring, aromatic ring may be optionally substituted by one or more halogen, -C ₁ to C ₃ alkyl, halogen-substituted -C ₁ to C ₃ alkyl, -OC ₁ to C ₃ alkyl; for example, R ¹ and R ⁵ are independently selected from hydrogen, methyl, ethyl, F, Cl, Br, I, -CF ₃ , -CH ₂ F, -CHF ₂ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ , -CH ₂ CF ₃ , -Cl, -Br, -OCH ₃ , Phenyl,

In certain specific embodiments, the present invention relates to compounds of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf), stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts thereof.

in,

R ² is selected from

y = 0, 1, 2 or 3;

wherein R ²¹ is selected from hydrogen, halogen, -OC ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl, -O-halogen substituted C ₁ ～C ₃ alkyl,

wherein R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, -C ₂ ～C ₄ alkenyl, -C ₂ ～C ₄ alkynyl, halogen-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₂ ～C ₄ alkenyl, halogen-substituted -C ₂ ～C ₄ alkynyl, -OC ₁ ～C ₃ alkyl (for example, R ² is specifically selected from );

L is selected from a bond, (For example, L is selected from a bond, Wherein the a end is connected to R ² ),

Wherein, R ⁴ and R ⁶ are independently selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl,

Wherein, R ⁴¹ is selected from hydrogen, halogen, methyl, ethyl, isopropyl (for example, L is specifically selected from a bond, For example, L is specifically selected from a bond, wherein the a end is connected to R ² );

R ^X and R ^X1 are independently selected from H, halogen, -C ₁ ～C ₃ alkyl, deuterium-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -C(O)CH ₃ ; for example, R ^X and R ^X1 are independently selected from H, -CD ₃ , -CH ₂ CHF ₂ , -Cl, -F, -CH ₃ , -CH ₂ F, -COCH ₃ ;

X ₄ respectively N, CR ⁵ ;

R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ to C ₃ alkyl, -C ₂ to C ₄ alkenyl, halogen-substituted -C ₁ to C ₃ alkyl, -OC ₁ to C ₃ alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring, aromatic ring may be optionally substituted by one or more halogen, -C ₁ to C ₃ alkyl, halogen-substituted -C ₁ to C ₃ alkyl, -OC ₁ to C ₃ alkyl; for example, R ¹ and R ⁵ are independently selected from hydrogen, methyl, ethyl, F, Cl, Br, I, -CF ₃ , -CH ₂ F, -CHF ₂ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ , -CH ₂ CF ₃ , -Cl, -Br, -OCH ₃ , Phenyl,

R ⁷ is selected from H, halogen, -CN, -NO ₂ , -NH ₂ , -C ₁ ～C ₃ alkyl, deuterium-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl; for example, R ⁷ is selected from H, methyl, ethyl, isopropyl, -Cl, -F;

R ³ is selected from H, halogen, -C ₁ -C ₃ alkyl, -C(O)NH ₂ , -C(O)NH(C ₁ -C ₃ alkyl), -C(O)N(C ₁ -C ₃ alkyl) ₂ ; for example, R ³ is selected from H, methyl, ethyl, isopropyl, -Cl, -F, -C(O)NHCH ₃ .

In certain specific embodiments, the inhibitory activity of the compounds of the present application on OCI-Ly19 cells is better than that of the compounds of the prior art. For example, in certain specific embodiments, the inhibitory activity of the compounds of the present application on OCI-Ly19 cells is 1 to 10 times that of the compounds of the prior art; for example, in certain specific embodiments, the inhibitory activity of the compounds of the present application on OCI-Ly19 cells is 10 to 20 times that of the compounds of the prior art; for example, in certain specific embodiments, the inhibitory activity of the compounds of the present application on OCI-Ly19 cells is 20 to 50 times that of the compounds of the prior art; for example, in certain specific embodiments, the inhibitory activity of the compounds of the present application on OCI-Ly19 cells is 50 to 100 times that of the compounds of the prior art; for example, in certain specific embodiments, the inhibitory activity of the compounds of the present application on OCI-Ly19 cells is 100 to 150 times or better than that of the compounds of the prior art.

Further, in certain specific embodiments, the compound of formula (I), its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts are selected from:

In the present application, the compounds of formula (I), formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf) defined in the above embodiments, their stereoisomers, deuterated forms, solvates, pharmaceutically acceptable salts, and the specific compounds disclosed in the examples of the present application are referred to as "compounds of the present invention".

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound of the present invention, its stereoisomers, deuterated substances, solvates, pharmaceutically acceptable salts, and pharmaceutically acceptable carriers and/or excipients.

The present invention also provides the use of the compound of the present invention, its stereoisomer, deuterated substance, solvate, pharmaceutically acceptable salt, or pharmaceutical composition thereof in the preparation of a drug for treating a PRMT5-mediated disease.

The present invention also provides the use of the compound of the present invention, its stereoisomer, deuterated substance, solvate, pharmaceutically acceptable salt, or pharmaceutical composition thereof in the preparation of a drug for treating diseases mediated by MTAP gene deletion and/or MTA accumulation.

The present invention also provides the compound of the present invention, its stereoisomer, deuterated substance, solvate, pharmaceutically acceptable salt, or pharmaceutical composition thereof, which is used for treating PRMT5-mediated diseases.

The present invention also provides the compound of the present invention, its stereoisomer, deuterated substance, solvate, pharmaceutically acceptable salt, or pharmaceutical composition thereof, which is used for treating diseases mediated by MTAP gene deletion and/or MTA accumulation.

Furthermore, the PRMT5-mediated disease and/or the MTAP gene deletion-mediated disease is cancer.

Diseases mediated by PRMT5 or mediated by MTAP gene deficiency and/or MTA accumulation deficiency as defined in the present invention include cancer or malignant tumors. "Cancer" or "malignant tumor" refers to any of a variety of diseases characterized by uncontrolled abnormal cell proliferation, the ability of affected cells to spread locally or to other parts of the body through the bloodstream and lymphatic system (i.e., metastasis), and a number of characteristic structural and/or molecular features. Any one of. "Cancer cell" refers to a cell that is in the early, middle or late stages of a multi-step tumor progression. The "cancer" or "malignant tumor" is selected from: neuroma, adenocarcinoma, adrenal cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendothelioma, hemangioma), appendix cancer, benign monoclonal gamma disease, bile duct cancer, bladder cancer, brain cancer (e.g., meningioma, glioma, such as astrocytoma, oligodendroglioma, medulloblastoma), bronchial cancer, carcinoid tumor, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colon cancer. Colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial cancer, ependymoma, endothelial sarcoma (e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma), endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarcinoma), Ewing's sarcoma, eye cancer (e.g., intraocular melanoma, retinoblastoma), hypereosinophilia, gallbladder cancer, gastric cancer (e.g., gastric adenocarcinoma), gastrointestinal stromal tumor (GIST), head and neck cancer (e.g., head Squamous cell carcinoma of the neck, oral cancer (e.g., oral squamous cell carcinoma, laryngeal cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer)), hematopoietic cancer (e.g., leukemia, such as acute lymphoblastic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myeloid leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myeloid leukemia (CML) (e.g., B-cell CML, T-cell CML), chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL), hairy cell leukemia (HCL), lymphomas such as follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), marginal zone B-cell lymphoma (e.g., mucosa-associated lymphoid tissue (MALT) lymphoma, lymph node marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt's lymphoma, lymphoplasmacytic lymphoma, immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma, and primary central nervous system (CNS) lymphoma; and non-Hodgkin lymphomas, such as precursor T lymphoblastic lymphoma/leukemia, peripheral T-cell lymphomas (such as cutaneous T-cell lymphoma (e.g., mycosis fungoides, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, mixture of one or more of the above leukemias/lymphomas; multiple myeloma (MM), heavy chain disease (e.g., alpha chain disease, gamma chain disease, μ chain disease), hemangioblastoma, inflammatory myofibroblastic tumor, immune cell amyloidosis, renal cancer (e.g., Wilms tumor, renal cell carcinoma), liver cancer (e.g., hepatocellular carcinoma, malignant hepatocellular carcinoma), lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), CLC), lung adenocarcinoma, leiomyosarcoma (LMS), mastocytosis (e.g., systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma, myeloproliferative disorders (MPD) (e.g., polycythemia vera (PV), essential thrombocythemia (ET), idiopathic extramedullary metaplasia of the bone marrow (AMM), chronic idiopathic myelofibrosis, chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES), neuroblastoma, neurofibromas (e.g., neurofibromatosis type 1 or type 2, schwannomatosis), neuroendocrine cancers (e.g., gastroenteropancreatic neuroendocrine tumors (GEP-NET), carcinoid tumors), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma, penile cancer, breast cancer, pancreatic cancer, skin cancer, Hodgkin lymphoma, cancers with homozygous deletion of the methylthioadenosine phosphorylase gene, neurofibrosarcoma.

Definition and Description

The compounds and derivatives provided in the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined in this document, the meaning that a person skilled in the art can give them should be given based on the disclosure and context.

The term "halogen" herein refers to F, Cl, Br, I or isotopes thereof.

The term "alkyl" refers to a saturated straight or branched aliphatic hydrocarbon group having 1 to 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) carbon atoms (i.e., _C1-20 alkyl). The alkyl group has 1 _to 12 carbon atoms (i.e., C1- _C12 alkyl) in certain specific embodiments, and 1 to 6 carbon atoms (i.e., _C1 - _C6 alkyl) in certain specific embodiments.

The term "alkenyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group containing at least one carbon-carbon double bond, which has 2 to 20 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) carbon atoms (i.e., _C2-20 alkenyl). In certain specific embodiments, the alkenyl group has 2 to 12 (e.g., ₂ , 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms (i.e., C2- _C12 alkenyl), and in certain specific embodiments, has 2 to 6 carbon atoms (i.e., _C2 - _C6 alkenyl).

The term "alkynyl" refers to a straight or branched unsaturated aliphatic hydrocarbon group containing at least one carbon-carbon triple bond and having 2 to 20 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) carbon atoms (i.e., _C2-20 alkynyl), which in certain specific embodiments has 2 to 12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms (i.e., _C2 - _C12 alkynyl), and in certain specific embodiments has 2 to 6 carbon atoms (i.e., _C2 - _C6 alkynyl).

The term "halogen-substituted alkyl" refers to a situation where one or more hydrogen atoms in an alkyl group are replaced by one or more halogen atoms (such as fluorine, chlorine, bromine, iodine or their isotopes). The upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be replaced in the alkyl group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit. Usually, the alkyl group is substituted by 1-5 halogens, or 1-3 halogens, or 1-2 halogens, or 1 halogen. When the number of halogen substituents is greater than 1, they can be substituted by the same or different halogens. Specific examples include, but are not limited to, _-CF3 , _-CH2Cl , _-CH2CF3 , _{-CF3 ,} etc.

The term "oxo" or "oxo group" refers to "=0" where an oxygen atom replaces two hydrogen atoms or a lone pair of electrons through a double bond.

In the "-C(O)R", "-S(O) _2R " and the like described in the present invention, the oxygen atom is connected to the carbon atom or the sulfur atom by a double bond, and the R group is connected to the carbon atom or the sulfur atom by a single bond. For another example, "-S(O)(NH)R" means that the oxygen atom and the nitrogen atom are connected to the sulfur atom by a double bond, and the R group is connected to the sulfur atom by a single bond.

Specific non-limiting examples of the term "-OC ₁ -C ₆ alkyl" include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexyloxy, cyclopropyloxy, cyclobutyloxy and the like.

The term "-O-halogen substituted C ₁ ～C ₆ alkyl" includes, but is not limited to, -O-halogenated C ₁ ～C ₆ alkyl, -O-halogenated C ₁ ～C ₄ alkyl or -O-halogenated C ₁ ～C ₂ alkyl; the upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be substituted by the substituted group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit. In some specific embodiments, the number of halogen substituents is 1-5 halogen substituents, 1-3 halogen substituents, or 1-5 halogen substituents. substituted, 1-2 halogen substituted, 1 halogen substituted; when the number of halogen substituents is greater than 1, they may be substituted by the same or different halogens; non-limiting examples include monofluoromethoxy, difluoromethoxy, trifluoromethoxy, difluoroethyloxy, and the like.

The term "carbocycle", "cycloalkyl" refers to a saturated or partially unsaturated monocyclic all-carbon ring (i.e., monocyclic cycloalkyl) or polycyclic all-carbon ring system (i.e., polycyclic cycloalkyl) having 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) ring atoms (i.e., 3 to 20-membered carbocycle or cycloalkyl). The cycloalkyl has 3 to 12 ring atoms (i.e., 3 to 12-membered carbocycle or cycloalkyl), 3 to 10 ring atoms (i.e., 3 to 10-membered carbocycle or cycloalkyl) in certain specific embodiments, 3 to 8 ring atoms (i.e., 3 to 8-membered carbocycle or cycloalkyl) in certain specific embodiments, 3 to 7 ring atoms (i.e., 3 to 7-membered carbocycle or cycloalkyl) in certain specific embodiments, and 3 to 6 ring atoms (i.e., 3 to 6-membered carbocycle or cycloalkyl) in certain specific embodiments.

The monocyclic cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl and cyclooctyl, etc. The polycyclic cycloalkyl includes spirocycloalkyl, condensed cycloalkyl and bridged cycloalkyl.

The term "spirocycloalkyl" refers to a full carbon polycyclic system with one carbon atom (called spiro atom) shared between the rings, which may contain one or more double bonds within the ring. The spirocycloalkyl has 6 to 14 ring atoms (i.e., 6 to 14-membered spirocycloalkyl) in certain specific embodiments, and 7 to 10 ring atoms (i.e., 7 to 10-membered spirocycloalkyl) in certain specific embodiments. Non-limiting examples include: Its connection point can be at any location.

The term "fused cycloalkyl" refers to a full-carbon polycyclic system that shares two adjacent carbon atoms between the rings, which is a monocyclic cycloalkyl fused to one or more monocyclic cycloalkyls and aryls, and may contain one or more double bonds in the ring, provided that the formed fused ring is non-aromatic. The fused cycloalkyl has 6 to 14 ring atoms (i.e., 6 to 14-membered fused cycloalkyl) in certain specific embodiments, and 7 to 10 ring atoms (i.e., 7 to 10-membered fused cycloalkyl) in certain specific embodiments. Non-limiting examples include: Its connection point can be at any location.

The term "bridged cycloalkyl" refers to a full-carbon polycyclic system that shares two carbon atoms that are not directly connected between the rings, and may contain one or more double bonds within the ring. The bridged cycloalkyl has 6 to 14 carbon atoms (i.e., 6 to 14-membered bridged cycloalkyl) in certain specific embodiments, and has 7 to 10 carbon atoms (i.e., 7 to 10-membered bridged cycloalkyl) in certain specific embodiments. The bridged cycloalkyl includes bicyclic bridged cycloalkyl and polycyclic The bridged cycloalkyl (such as a tricyclic bridged cycloalkyl, a tetracyclic bridged cycloalkyl, etc.), in certain specific embodiments, is a bicyclic bridged cycloalkyl or a tricyclic bridged cycloalkyl. Non-limiting examples include: Its connection point can be at any location.

The term "heterocycle" or "heterocyclyl" refers to a saturated or non-aromatic partially saturated monocyclic heterocycle (i.e., a monocyclic heterocyclyl) or a polycyclic heterocyclic ring system (i.e., a polycyclic heterocyclyl), which contains at least one (e.g., 1, 2, 3 or 4) heteroatoms selected from nitrogen, oxygen and sulfur (the nitrogen may be optionally oxidized, i.e., to form nitrogen oxides, or quaternized; the sulfur may be optionally oxoed, i.e., to form sulfoxides or sulfones) in the ring. Non-limiting examples of the monocyclic heterocyclyl include: The polycyclic heterocycle also includes a heterocycle fused with one or more of a carbocycle, a heterocycle, an aryl group or a heteroaryl group to form a fused heterocycle, the attachment site of which may be located at a non-aromatic carbon atom, an aromatic carbon atom or a heteroatom. Non-limiting examples of the fused heterocycle include The polycyclic heterocyclic group also includes a spiro heterocyclic group and a bridged heterocyclic group.

The term "spiro hetero" or "spiroheterocyclyl" refers to a polycyclic heterocyclic ring system in which the rings share one atom (called a spiro atom), which may contain one or more double bonds in the rings, and which contains at least one (e.g., 1, 2, 3 or 4) heteroatoms selected from nitrogen, oxygen and sulfur in the rings (the nitrogen may be optionally oxidized, i.e., to form nitrogen oxides, or quaternized; the sulfur may be optionally oxoed, i.e., to form sulfoxides or sulfones), non-limiting examples include: wait.

The term "bridged hetero" or "bridged heterocyclic group" refers to a polycyclic heterocyclic ring system that shares two atoms that are not directly connected between the rings, which may contain one or more double bonds in the ring and at least one (e.g., 1, 2, 3 or 4) heteroatom selected from nitrogen, oxygen and sulfur in the ring (the nitrogen may be optionally oxidized, i.e., to form nitrogen oxides, or quaternized; the sulfur may be optionally oxoed, i.e., to form sulfoxides or sulfones), non-limiting examples include: wait.

The term "aromatic ring" or "aryl" refers to a monocyclic all-carbon aromatic ring (i.e., monocyclic aromatic group) or a polycyclic aromatic ring system (i.e., polycyclic aromatic group) having a conjugated π electron system, wherein the aromatic group has 6 to 10 ring atoms (i.e., 6 to 10-membered aromatic group) in certain specific embodiments. Examples of the monocyclic aromatic group include phenyl. Non-limiting examples of the polycyclic aromatic group include naphthyl, anthracenyl, phenanthrenyl, etc.

The terms "heteroaromatic ring" and "heteroaryl" refer to a monocyclic heteroaromatic ring (i.e., a monocyclic heteroaryl) or a polycyclic heteroaromatic ring system (i.e., a polycyclic heteroaryl) having a conjugated π electron system, which contains at least one (e.g., 1, 2, 3 or 4) heteroatoms selected from nitrogen, oxygen and sulfur in the ring (the nitrogen may be optionally oxidized, i.e., to form nitrogen oxides, or quaternized; the sulfur may be optionally oxoed, i.e., to form sulfoxides or sulfones), and the heteroaryl has 5 to 10 ring atoms (i.e., a 5- to 10-membered heteroaryl) in certain specific embodiments, and 5 or 6 ring atoms (i.e., a 5- or 6-membered heteroaryl) in certain specific embodiments. The monocyclic heteroaryl groups include, but are not limited to, furanyl, thienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, furazanyl, pyrrolyl, N-alkylpyrrolyl, pyridyl, pyrimidinyl, pyridonyl, pyrazinyl, pyridazinyl, etc. The polycyclic heteroaryl groups include, but are not limited to, indolyl, indazolyl, quinolyl, isoquinolyl, quinoxalinyl, phthalazinyl, benzimidazolyl, benzothienyl, quinazolinyl, benzothiazolyl, carbazolyl, etc.

The term "optional" or "optionally" means that the event or circumstance described subsequently may but need not occur, and the description includes occasions where the event or circumstance occurs or does not occur. For example, "alkyl optionally substituted with F" means that alkyl may but need not be substituted with F, and the description includes the situation where alkyl is substituted with F and the situation where alkyl is not substituted with F.

The group description of the present invention Used to describe the substitution position of a group, for example, The tetrahydropyrrole ring passes through The position forms a spiral ring with other rings in the structure.

The term "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention which retains the biological effectiveness and properties of the free acid or free base and which is obtained by reacting the free acid with a non-toxic inorganic or organic base or the free base with a non-toxic inorganic or organic acid.

The term "pharmaceutical composition" refers to a mixture of one or more compounds described herein, their stereoisomers, deuterated forms, solvates, pharmaceutically acceptable salts, and other components, wherein the other components include physiologically/pharmaceutically acceptable carriers and/or excipients.

The term "carrier" refers to a system that does not cause significant irritation to the organism and does not eliminate the biological activity and properties of the administered compound, and can change the way the drug enters the human body and its distribution in the body, control the release rate of the drug and deliver the drug to the targeted organ. Non-limiting examples include microcapsules and microspheres, nanoparticles, liposomes, etc.

The term "excipient" refers to a substance that is not a therapeutic agent in itself but is used as a diluent, adjuvant, binder and/or vehicle for addition to a pharmaceutical composition to improve its handling or storage properties or to allow or facilitate the formation of a compound or pharmaceutical composition into a unit dosage form for administration. As known to those skilled in the art, pharmaceutical excipients can provide a variety of functions and can be described as wetting agents, buffers, suspending agents, lubricants, emulsifiers, disintegrants, absorbents, preservatives, surfactants, colorants, flavoring agents and sweeteners.

The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, diastereomers and conformational isomers.

The term "solvate" refers to a substance formed by a stoichiometric or non-stoichiometric amount of a solvent that is bound to the compound or salt of the present invention by non-covalent forces between molecules. When the solvent is water, it is a hydrate.

The term "deuterated compound" refers to a molecule or group in which one or more hydrogen atoms are replaced by deuterium atoms, wherein the proportion of deuterium atoms is greater than the abundance of deuterium in nature.

DETAILED DESCRIPTION

The present invention is further explained below in conjunction with specific examples, but the examples do not limit the present invention in any form.

Abbreviation Description:

PMB denotes p-methoxybenzyl; Boc denotes tert-butyloxycarbonyl; DMAP denotes 4-dimethylaminopyridine; Na ₂ CO ₃ denotes sodium carbonate; THF denotes tetrahydrofuran; DBU denotes 1,8-diazabicyclo[5.4.0]undec-7-ene; AcOH denotes acetic acid; Pd(dppf)Cl ₂ denotes 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) dichloride; K ₂ CO ₃ denotes potassium carbonate; SEMCl denotes 2-(trimethylsilyl)ethoxymethyl chloride; RT/rt denotes room temperature; LiAlH ₄ denotes lithium aluminum hydride; DEAD denotes diethyl azodicarboxylate; PPh ₃ denotes triphenylphosphine; NaBH(OAc) ₃ represents sodium triacetoxyborohydride; EDCI represents 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; Py represents pyridine; TFAA represents trifluoroacetic anhydride; TEA represents triethylamine; DIEA represents N,N-diisopropylethylamine; t-Bubrettphos represents 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'triisopropylbiphenyl; NMP represents N-methylpyrrolidone; Pd ₂ (dba) ₃ represents tris(dibenzylideneacetone)dipalladium; Xantphos represents 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene; NaBH ₃ CN represents sodium cyanoborohydride; HATU represents 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate; Pd(OAc) ₂ represents palladium acetate; X-Phos represents 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl; Cs ₂ CO ₃ represents cesium carbonate; DIAD represents diisopropyl azodicarboxylate; MeONa represents sodium methoxide; MeOH represents methanol; KOAc represents potassium acetate; DMSO represents dimethyl sulfoxide; NaBH ₃ CN represents sodium cyanoborohydride; HBr represents hydrobromic acid; TBAB represents tetrabutylammonium bromide; LiBr represents lithium bromide; DMAP represents 4-dimethylaminopyridine; NIS represents N-iodosuccinimide; NBS represents N-bromosuccinimide; TFA represents trifluoroacetic acid; DCM represents dichloromethane; TCFH represents N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate; NMI represents N-methylimidazole; Pd(PPh ₃ ) ₄ represents tetrakistriphenylphosphine palladium; K ₃ PO ₄ represents potassium phosphate; CuI represents cuprous iodide; t-BuOK represents potassium tert-butoxide; DMF represents N,N-dimethylformamide; ACN represents acetonitrile; TMEDA represents tetramethylethylenediamine; n-BuLi represents n-butyllithium; m-CPBA represents m-chloroperbenzoic acid; POCl ₃ represents phosphorus oxychloride; DPPA represents diphenylphosphoryl azide; HOBt represents 1-hydroxybenzotriazole; Brettphos-G3-Pd represents methanesulfonate (2-dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium; ZnBr ₂ represents zinc bromide; Pd(PPh ₃ ) ₂ Cl ₂ represents bis(triphenylphosphine)palladium dichloride; Brettphos represents 2-(dicyclohexylphosphine)3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl.

Preparation of intermediates

Preparation Example 1: Preparation of (5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methanol

Step 1: Preparation of 3-bromo-N,N-bis[(4-methoxyphenyl)methyl]-6-methyl-5-nitropyridin-2-amine

3-Bromo-2-chloro-6-methyl-5-nitropyridine (12 g, 47.72 mmol) and bis[(4-methoxyphenyl)methyl]amine (14.74 g, 57.26 mmol) were dissolved in tetrahydrofuran (60 mL), sodium carbonate (6.07 g, 57.26 mmol) was added, and the mixture was reacted at 75°C for 16 hours. After LC-MS showed that the reaction was complete, the system was filtered and concentrated, and purified and separated by silica gel column to obtain 11.7 g of the title compound.

MS (ESI) m/z (M+H) ⁺ = 472.1.

Step 2: Preparation of ethyl 3-(6-(bis[(4-methoxyphenyl)methyl]amino)-5-bromo-3-nitropyridin-2-yl)-2-oxopropanoate

Weigh 3-bromo-N,N-bis[(4-methoxyphenyl)methyl]-6-methyl-5-nitropyridine-2-amine (11.7 g, 24.77 mmol), dissolve in diethyl oxalate (10.86 g, 74.31 mmol), add 1,8-diazabicycloundec-7-ene (4.53 g, 29.72 mmol) dropwise at 40°C. React at 40°C for 12 hours. LC-MS shows that a small amount of raw material remains. Water is added to the system, and ethyl acetate is extracted three times. The organic phase is concentrated and purified by silica gel column to obtain 13 g of the title compound as a light white solid.

MS (ESI) m/z (M+H) ⁺ = 572.1.

Step 3: Preparation of ethyl 5-(bis(4-methoxybenzyl)amino)-6-bromo-1H-pyrrolo[3,2-b]pyridine-2-carboxylate

Weigh 3-(6-(bis[(4-methoxyphenyl)methyl]amino)-5-bromo-3-nitropyridin-2-yl)-2-oxopropanoic acid ethyl ester (13g, 22.71mmol), dissolve in acetic acid (150mL), add iron powder (5.07g, 90.84mmol), and react at room temperature for 12 hours. LC-MS shows that the reaction is complete, water is added to the system, and ethyl acetate is extracted three times, washed with saturated sodium bicarbonate solution, the organic phase is concentrated, and purified on a silica gel column to obtain 8.5g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 524.1.

Step 4: Preparation of ethyl 5-(bis(4-methoxybenzyl)amino)-6-methyl-1H-pyrrolo[3,2-b]pyridine-2-carboxylate

Weigh 5-(bis(4-methoxybenzyl)amino)-6-bromo-1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid ethyl ester (8.5 g, 16.21 mmol) and dissolve it in 1,4-dioxane (80 mL), then add methylboric acid (3.88 g, 64.84 mmol), dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (0.59 g, 0.81 mmol), potassium carbonate (8.96 g, 64.84 mmol) in sequence, replace the nitrogen three times, heat to 110°C and react for 12 hours. After LC-MS shows that the reaction is complete, filter, concentrate the filtrate, and purify on a silica gel column to obtain 6 g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 460.2.

Step 5: Preparation of ethyl 5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridine-2-carboxylate

Dissolve 5-(bis(4-methoxybenzyl)amino)-6-methyl-1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid ethyl ester (6g, 13.06mmol) in tetrahydrofuran (60mL), add sodium hydride (0.41g, 16.98mmol) in batches under ice-water bath, stir for 20 minutes, add 2-(trimethylsilyl)ethoxymethyl chloride (2.83g, 16.98mmol), and react at room temperature for 2 hours. After TLC shows that the reaction is complete, add water to quench, extract three times with ethyl acetate, combine the organic phases and backwash once with saturated brine, dry over anhydrous sodium sulfate, and purify on a silica gel column to obtain 7.5g of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 590.3.

Step 6: Preparation of (5-(bis[(4-methoxyphenyl)methyl]amino)-6-methyl-1-[(2-(trimethylsilyl)ethoxy)methyl]-1H-pyrrolo[3,2-b]pyridin-2-yl)methanol

Dissolve 5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid ethyl ester (7.5 g, 12.72 mmol) in tetrahydrofuran (100 mL), add lithium aluminum hydride (0.72 g, 19.08 mmol) in portions under ice-water bath, and gradually add 2-nitropropene. The mixture was heated to room temperature and reacted for 1 hour. LC-MS showed that the reaction was complete. Water (1 mL) was added under ice-water bath, and stirred for 5 minutes. 15% sodium hydroxide solution (1 mL) was added dropwise, and then water (3 mL) was slowly added dropwise. The mixture was stirred at room temperature for 0.5 hour, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column to obtain 6.8 g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 548.3.

Preparation Example 2: Preparation of 1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-oxo-1,6-dihydropyridine-2-carboxylic acid

Step 1: Preparation of methyl 1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-oxo-1,6-dihydropyridine-2-carboxylate

(5-(bis[(4-methoxyphenyl)methyl]amino)-6-methyl-1-[(2-(trimethylsilyl)ethoxy)methyl]-1H-pyrrolo[3,2-b]pyridin-2-yl)methanol (2g, 3.56mmol) and methyl 6-oxo-1,6-dihydropyridine-2-carboxylate (0.84g, 5.47mmol) were dissolved in tetrahydrofuran (30mL), triphenylphosphine (1.44g, 5.47mmol) was added under ice-water bath, diethyl azodicarboxylate (0.95g, 5.47mmol) was added dropwise, and stirred at room temperature for 12 hours. After LC-MS showed that the reaction was complete, water was added to quench, and ethyl acetate was extracted three times. The organic phases were combined and backwashed once with saturated brine, dried over anhydrous sodium sulfate, and purified on a silica gel column to obtain 0.447g of the title compound as an oil.

MS (ESI) m/z (M+H) ⁺ = 683.3.

Step 2: Preparation of 1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-oxo-1,6-dihydropyridine-2-carboxylic acid

1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-oxo-1,6-dihydropyridine-2-carboxylic acid methyl ester (447 mg, 0.65 mmol) was dissolved in tetrahydrofuran (5 mL) and water (5 mL), and monohydrate was added. After adding lithium hydroxide (54.55 mg, 1.3 mmol), the system was reacted at room temperature for 1 hour. After LC-MS showed that the reaction was complete, the system was concentrated, purified by reverse phase preparation, and lyophilized to obtain 380 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 669.3.

Preparation Example 3: Preparation of N-isopropyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of N-isopropylaniline

Aniline (500.0 mg, 5.37 mmol) and propan-2-one (1559.4 mg, 26.85 mmol) were dissolved in tetrahydrofuran (20 mL), and sodium triacetoxyborohydride (2276.2 mg, 10.74 mmol) was added at room temperature. After the addition, the mixture was reacted at room temperature for 12 hours. After TLC showed that the reaction was complete, water was added to quench the reaction, and the mixture was extracted several times with dichloromethane and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane/methanol = 15/1 (V:V)) to obtain 350.0 mg of the title compound as a light yellow oil.

MS (ESI) m/z (M+H) ⁺ = 136.2.

Step 2: Preparation of N-isopropyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Dissolve N-isopropylaniline (350.0 mg, 2.59 mmol) and 6-oxo-1,6-dihydropyridine-2-carboxylic acid (360.2 mg, 2.59 mmol) in pyridine (5 mL); add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (993.0 mg, 5.18 mmol) at room temperature, and react at room temperature for 2 hours. After TLC shows that the reaction is complete, concentrate the solvent under reduced pressure to obtain a crude product, which is purified by column chromatography (dichloromethane/methanol = 10/1 (V:V)) to obtain 300.0 mg of the title compound as a light green solid.

MS (ESI) m/z (M+H) ⁺ = 257.3.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 11.51 (s, 1H), 7.40–7.24 (m, 3H), 7.26–7.10 (m, 3H), 6.21–5.91 (m, 2H), 4.90–4.71 (m, 1H), 1.10 (d, J = 6.8Hz, 6H).

Preparation Example 4: Preparation of N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

The title compound was prepared using the corresponding commercial reagents as raw materials and a preparation method similar to that of the above Preparation Example 3.

MS (ESI) m/z (M+H) ⁺ = 229.2.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 11.51 (s, 1H), 7.34 (t, J = 7.6 Hz, 2H), 7.29–7.18 (m, 4H), 6.25 (d, J = 9.0 Hz, 1H), 6.08 (s, 1H), 3.33 (s, 3H).

Preparation Example 5: Preparation of tert-butyl (2-(hydroxymethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate

Step 1: Preparation of ethyl 5-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridine-2-carboxylate

Dissolve 5-chloro-1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid ethyl ester (1g, 4.45mmol) in tetrahydrofuran (30mL), add sodium hydride (0.14g, 5.79mmol) in batches under ice-water bath, stir at this temperature for 30 minutes, add 2-(trimethylsilyl)ethoxymethyl chloride (0.96g, 5.79mmol), and react at room temperature for 2 hours. After TLC shows that the reaction is complete, add water to quench, extract with ethyl acetate three times, combine the organic phases, backwash once with saturated brine, dry over anhydrous sodium sulfate, and purify on a silica gel column to obtain 1.5g of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 355.1.

Step 2: Preparation of ethyl 5-((tert-butoxycarbonyl)amino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridine-2-carboxylate

5-Chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid ethyl ester (500 mg, 1.1 mmol) and tert-butyl carbamate (330 mg, 2.82 mmol) were dissolved in 1,4-dioxane solution (20 mL), palladium acetate (31.66 mg, 0.14 mmol), 2-(dicyclohexylphosphino)-2,4,6-triisopropylbiphenyl (134.4 mg, 0.28 mmol) and cesium carbonate (0.92 mg, 2.82 mmol) were added, and the reaction system was stirred for 2 hours. The reaction was carried out at 100° C. for 12 hours. LC-MS showed that the reaction was complete, the system was filtered through celite, the filtrate was concentrated to dryness, and purified by silica gel column to obtain 0.46 g of the title compound as a light white solid.

MS (ESI) m/z (M+H) ⁺ = 436.2.

Step 3: Preparation of tert-butyl (2-(hydroxymethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate

Dissolve 5-((tert-butoxycarbonyl)amino)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid ethyl ester (460 mg, 1.06 mmol) in tetrahydrofuran (40 mL), add lithium aluminum hydride (60.3 mg, 1.59 mmol) in batches under ice-water bath, stir the system for 1 hour, and monitor the reaction completion by LC-MS. Add water (1 mL) under ice-water bath, stir for 5 minutes, add 15% sodium hydroxide solution (1 mL) dropwise, then slowly add water (3 mL), stir at room temperature for 0.5 hour, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and purify on silica gel column to obtain 320 mg of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 294.2.

Preparation Example 6: Preparation of 1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-bromopyridin-2(1H)-one

(5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methanol (1.0 g, 1.83 mmol), triphenylphosphine (718.3 mg, 2.74 mmol) and 6-bromopyridin-2(1H)-one (349.4 mg, 2.01 mmol) were dissolved in tetrahydrofuran (50 mL), and then diisopropyl azodicarboxylate (553.7 mg, 2.74 mmol) was slowly added dropwise under nitrogen protection in an ice-water bath. After the addition was completed, the reaction system was stirred at room temperature for 3 hours. LCMS showed that the reaction was complete. The reaction solution was concentrated and separated and purified by a chromatography column to obtain 350.0 mg of the yellow gum title compound.

MS (ESI) m/z (M+H) ⁺ = 703.3.

Preparation Example 7: 4-Methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrrolo[2,1-f][1,2,4]triazine

Step 1: Preparation of 5-bromo-4-methoxypyrrolo[2,1-f][1,2,4]triazine

5-Bromo-4-chloropyrrolo[2,1-f][1,2,4]triazine (0.5 g, 2.15 mmol) was dissolved in anhydrous methanol (10 mL), followed by the addition of sodium methoxide (174.3 mg, 3.23 mmol). After the addition was complete, the reaction system was stirred at room temperature for 16 hours. LCMS showed that the reaction was complete, the reaction solution was added with water, filtered, and the filter cake was dried to obtain the title compound (450.0 mg, crude product) as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 228.1.

Step 2: Preparation of 4-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrrolo[2,1-f][1,2,4]triazine

5-Bromo-4-methoxypyrrolo[2,1-f][1,2,4]triazine (410.0 mg, 1.80 mmol), biboronic acid pinacol ester (730.5 mg, 2.88 mmol) and potassium acetate (705.8 mg, 7.19 mmol) were dissolved in tetrahydrofuran (14.0 mL) and dimethyl sulfoxide (2.0 mL), and then [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (130.5 mg, 0.18 mmol) was added. After the addition was completed, the reaction system was placed in a nitrogen atmosphere and heated to 80°C and stirred for 16 hours. LCMS showed that the raw materials were basically reacted. After the reaction solution was concentrated, it was separated and purified by column chromatography to obtain 400.0 mg of the title compound as a light yellow oil.

MS (ESI) m/z (M+H) ⁺ = 276.2.

Preparation Example 8: Preparation of 6-((3,4-dihydroquinolin-1(2H)-yl)methyl)-1-(prop-2-yn-1-yl)pyridin-2(1H)-one

Step 1: Preparation of 1-((6-methoxypyridin-2-yl)methyl)-1,2,3,4-tetrahydroquinoline

To a dry round-bottom flask, add 1,2,3,4-tetrahydroquinoline (2.9 g, 21.77 mmol), 6-methoxypyridine-2-carboxaldehyde (4.5 g, 32.66 mmol), and methanol (50 mL) in sequence. After stirring evenly, slowly add sodium cyanoborohydride (2.1 g, 32.66 mmol) at room temperature, and then react the reaction solution at room temperature for 5 hours. LC-MS monitors the reaction, and a small amount of raw materials remain. Add water to quench the reaction, extract with ethyl acetate three times, and combine the organic phases. The organic phase was backwashed once with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was spin-dried. The crude product was purified by column chromatography to obtain 3.2 g of the title product as a yellow viscous liquid.

MS (ESI) m/z (M+H) ⁺ = 255.1.

Step 2: Preparation of 6-((3,4-dihydroquinolin-1(2H)-yl)methyl)pyridin-2(1H)-one

1-[(6-methoxypyridin-2-yl)methyl]-1,2,3,4-tetrahydroquinoline (1.5 g, 5.90 mmol), hydrobromic acid (5 mL, 7.45 g, 92.08 mmol) and acetic acid (10 mL) were added to the reaction tube in sequence. After stirring evenly, the reaction system was placed in an 80°C oil bath for 6 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 1.3 g of the title product as a light green solid.

MS (ESI) m/z (M+H) ⁺ = 241.1.

Step 3: Preparation of 6-((3,4-dihydroquinolin-1(2H)-yl)methyl)-1-(prop-2-yn-1-yl)pyridin-2(1H)-one

6-[(1,2,3,4-tetrahydroquinolin-1-yl)methyl]-1,2-dihydropyridin-2-one (600.0 mg, 2.50 mmol), potassium carbonate (1.1 g, 7.50 mmol), tetrabutylammonium bromide (890.0 g, 0.28 mmol) and lithium bromide (430.0 mg, 5.00 mmol) were added to the reaction tube in sequence, followed by toluene (40 mL) and water (1 mL). After stirring evenly, 3-bromopropyne (480.0 mg, 4.00 mmol) was slowly added. After the reaction solution was reacted at 80 ° C for 6 hours, LC-MS monitored the reaction to be complete. Water was added to quench the reaction, extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 630.0 mg of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 279.1.

Preparation Example 9: Preparation of tert-butyl N-(5-amino-6-iodo-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Step 1: Preparation of tert-butyl N-[(tert-butoxy)carbonyl]-N-(3-methyl-5-nitropyridin-2-yl)carbamate

Weigh 3-methyl-5-nitropyridine-2-amine (9.0 g, 58.8 mmol) and dissolve it in dichloromethane (100 mL). Add di-tert-butyl dicarbonate (12.8 g, 58.8 mmol) and 4-dimethylaminopyridine (7.2 g, 16.0 mmol) in an ice-water bath. After the addition, react at room temperature for 3 hours. After TLC shows that the reaction is complete, add the system dropwise into (500 mL) water in an ice-water bath, extract three times with ethyl acetate, combine the organic phases and react with saturated brine. The residue was washed once, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 20.0 g of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 354.1.

Step 2: Preparation of tert-butyl N-(5-amino-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Tert-butyl N-[(tert-butoxy)carbonyl]-N-(3-methyl-5-nitropyridin-2-yl)carbamate (20.0 g, 56.7 mmol) was dissolved in methanol (200 mL), and palladium carbon (4.0 g, 10% (W/W)) was slowly added in batches at 0°C. After the addition, the hydrogen was replaced three times, and then the reaction was carried out under a hydrogen atmosphere at room temperature for 10 hours. TLC showed that the raw material was completely consumed. The system was filtered and concentrated to obtain 18.0 g of the title compound as a colorless oil, which was directly used in the next step reaction.

MS (ESI) m/z (M+H) ⁺ = 324.1.

Step 3: Preparation of tert-butyl N-(5-amino-6-iodo-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Dissolve tert-butyl N-(5-amino-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (18.0 g, 55.7 mmol) in N,N-dimethylformamide (100 mL), add N-iodosuccinimide (12.5 g, 55.7 mmol) at 0°C, and return to room temperature to react for 1 hour. After TLC shows that the reaction is complete, add (1 L) water to the system, extract with ethyl acetate five times, combine the organic phases and backwash once with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The crude product is purified by column chromatography (dichloromethane/methanol = 20/1 (V:V)) to obtain 15.0 g of the title compound as a dark green solid.

MS (ESI) m/z (M+H) ⁺ = 450.1.

Preparation Example 10: Preparation of N-((5-fluoropyridin-2-yl)methyl)-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of N-((5-fluoropyridin-2-yl)methyl)aniline

Dissolve 5-fluoropyridine-2-carboxaldehyde (500 mg, 4 mmol) and aniline (745.04 mg, 8 mmol) in methanol (10 mL), add 3 drops of acetic acid solution, and stir at room temperature for 20 minutes. Add sodium cyanoborohydride (502.72 mg, 8 mmol) in batches under an ice-water bath, stir at room temperature for 4 hours, and monitor the reaction by LC-MS. Add water to quench the reaction, extract three times with ethyl acetate, combine the organic phases, backwash once with saturated brine, dry over anhydrous sodium sulfate, and purify on a silica gel column to obtain 650 mg of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 203.1.

Step 2: Preparation of N-((5-fluoropyridin-2-yl)methyl)-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Weigh 6-oxo-1,6-dihydropyridine-2-carboxylic acid (446.54 mg, 3.21 mmol) and dissolve it in pyridine (20 mL), then add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (738.43 mg, 3.85 mmol) and N-((5-fluoropyridin-2-yl)methyl)aniline (650 mg, 3.21 mmol) in sequence. The system reacts at room temperature for 12 hours. After LC-MS shows that the reaction is complete, the system is concentrated, the acid is adjusted, and reverse phase purification is performed to obtain 600 mg of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 324.1.

Preparation Example 11: Preparation of N-benzyl-N-[(5-fluoropyridin-2-yl)methyl]-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of N-benzyl-1-(5-fluoropyridin-2-yl)methanamine

To a dry round-bottom flask, 5-fluoropyridine-2-carboxaldehyde (1.0 g, 7.99 mmol), benzylamine (1.71 g, 15.98 mmol) and methanol (10 mL) were added in sequence. Sodium cyanoborohydride (1.2 g, 19.09 mmol) was slowly added at room temperature, and then the reaction solution was reacted at room temperature for 2 hours. LC-MS monitoring The reaction was tested, and a small amount of raw material remained. Water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was dried by spin drying. The crude product was purified by column chromatography to obtain 1.2 g of the title product as a yellow oil.

MS (ESI) m/z (M+H) ⁺ = 217.2.

Step 2: Preparation of N-benzyl-N-[(5-fluoropyridin-2-yl)methyl]-6-oxo-1,6-dihydropyridine-2-carboxamide

To the reaction tube, add 6-oxo-1,6-dihydropyridine-2-carboxylic acid (0.85g, 6.11mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.60g, 8.32mmol) and pyridine (10mL) in sequence; after the addition, stir evenly, add N-benzyl-1-(5-fluoropyridin-2-yl)methylamine (1.2g, 5.55mmol) dropwise, and then place the reaction system at room temperature for 6 hours. After the reaction is completed, add water to quench the reaction, extract with ethyl acetate three times, dry with anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by silica gel column to obtain 1.3g of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 338.1.

Step 3: Preparation of N-benzyl-N-[(5-fluoropyridin-2-yl)methyl]-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Into the reaction tube, N-benzyl-N-[(5-fluoropyridin-2-yl)methyl]-6-oxo-1,6-dihydropyridine-2-carboxamide (1.24g, 3.68mmol), potassium carbonate (1.53g, 11.04mmol), lithium bromide (0.64g, 7.36mmol), tetrabutylammonium bromide (0.13g, 0.4mmol), toluene (20mL) and water (1mL) were added in sequence. After stirring evenly, 3-bromopropyne (0.66g, 5.52mmol) was slowly added at room temperature. The reaction system was placed in an 80°C oil bath for 6 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 0.6g of the title product.

MS (ESI) m/z (M+H) ⁺ = 376.1.

Preparation Example 12: Preparation of tert-butyl (tert-butoxycarbonyl)(6-iodo-3-methyl-5-(2,2,2-trifluoroacetylamino)pyridin-2-yl)carbamate

Dissolve tert-butyl (5-amino-6-iodo-3-methylpyridin-2-yl)(tert-butoxycarbonyl)carbamate (4.493 g, 10 mmol) in dichloromethane (100 mL), add trifluoroacetic anhydride (1.67 mL, 12 mmol) and triethylamine (2.78 mL, 20 mmol) at room temperature, and react at room temperature for 2 hours. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, extracted several times with dichloromethane, and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography to obtain 5.18 g of the title compound.

MS (ESI) m/z (M+H) ⁺ = 546.1.

Preparation Example 13: Preparation of (2-(4-fluorophenyl)pyrrolidin-1-yl)(6-hydroxy-4-methylpyridin-2-yl)methanone

Step 1: Preparation of (6-chloro-4-methylpyridin-2-yl)(2-(4-fluorophenyl)pyrrolidin-1-yl)methanone

6-Chloro-4-methylpicolinic acid (550.0 mg, 3.22 mmol) was dissolved in dichloromethane (10 mL). N, N-diisopropylethylamine (830.0 mg, 6.44 mmol) and 2-(7-azobenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (1.2 g, 3.22 mmol) were added in sequence under ice bath conditions. Finally, 2-(4-fluorophenyl)pyrrolidine (531 mg, 3.22 mmol) was added. After the addition, the mixture was reacted at room temperature for 4 hours. After TLC showed that the reaction was complete, 200 mL of water was added to the system, and the mixture was extracted with ethyl acetate three times. The organic phases were combined and backwashed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 960 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 319.1.

Step 2: Preparation of (2-(4-fluorophenyl)pyrrolidin-1-yl)(6-hydroxy-4-methylpyridin-2-yl)methanone

Weigh (6-chloro-4-methylpyridin-2-yl)(2-(4-fluorophenyl)pyrrolidin-1-yl)methanone (960 mg, 3.02 mmol), boric acid (276.0 mg, 4.53 mmol), palladium acetate (68.0 mg, 0.30 mmol), 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'tri-isopropyl-1,1'-biphenyl (293.0 mg, 0.60 mmol), cesium carbonate (2.0 g, 6.04 mmol), then seal the system for nitrogen protection, add solvent N-methylpyrrolidone (10 mL) to dissolve, then ventilate the system three times with nitrogen, then react at 90°C for 3 hours, and monitor the reaction of the raw materials by LCMS. The system is quenched with water, extracted with ethyl acetate, concentrated, and the crude product is separated by chromatography column (dichloromethane/methanol=20/1) to obtain 400 mg of the title compound as a purple-black solid.

MS (ESI) m/z (M+H) ⁺ = 301.1.

Preparation Example 14: Preparation of 6-chloro-5-fluoro-2-iodopyridin-3-amine

Step 1: Preparation of tert-butyl N-(6-chloro-5-fluoropyridin-3-yl)carbamate

5-Bromo-2-chloro-3-fluoropyridine (4.0 g, 19.01 mmol) was dissolved in 1,4-dioxane (80 mL), and tert-butyl carbamate (2.6 g, 22.81 mmol), tris(dibenzylideneacetone)dipalladium (1.7 g, 1.90 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (2.2 g, 3.80 mmol) and cesium carbonate (12.4 g, 38.02 mmol) were added at room temperature, and the reaction was carried out at 90°C for 8 hours after replacing the inert gas. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, and the mixture was extracted several times with ethyl acetate and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (petroleum ether/ethyl acetate = 5/1 (V:V)) to obtain 2.0 g of the title compound as a light yellow oil.

MS (ESI) m/z (M+H) ⁺ = 247.7.

Step 2: Preparation of 6-chloro-5-fluoropyridin-3-amine

Dissolve tert-butyl N-(6-chloro-5-fluoropyridin-3-yl)carbamate (1.9 g, 7.70 mmol) in a solution of hydrochloric acid (0.6 g, 15.02 mmol) in 1,4-dioxane (2 mL). After addition, react at room temperature for 2 hours. After TLC shows that the reaction is complete, concentrate the solvent, add sodium bicarbonate to the reaction system to adjust the system to neutrality, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (dichloromethane/methanol = 10/1 (V:V)) to obtain 1.0 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ =147.5.

Step 3: Preparation of 6-chloro-5-fluoro-2-iodopyridin-3-amine

Dissolve 6-chloro-5-fluoropyridin-3-amine (1.0 g, 6.82 mmol) in N,N-dimethylformamide (10 mL), add N-iodosuccinimide (3.1 g, 13.64 mmol) at room temperature, and react at room temperature for 8 hours. After TLC shows that the reaction is terminated, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (dichloromethane/methanol = 15/1 (V:V)) to obtain 0.2 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 273.5.

Preparation Example 15: Preparation of N-methyl-6-oxo-N-phenyl-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Dissolve N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide (1.1 g, 4.82 mmol) in N,N-dimethylformamide (15 mL), add 3-bromopropyne (1.2 g, 9.64 mmol) and potassium carbonate (1.4 g, 9.64 mmol) at room temperature, and react at 50°C for 2 hours. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (dichloromethane/methanol = 20/1 (V:V)) to obtain 0.8 g of the title compound as a light yellow oil.

MS (ESI) m/z (M+H) ⁺ = 267.2.

Preparation Example 16: Preparation of 4-methyl-6-(2-phenylpyrrolidine-1-carbonyl)-1-(prop-2-yn-1-yl)pyridin-2(1H)-one

Step 1: Preparation of (6-chloro-4-methylpyridin-2-yl)(2-phenylpyrrolidin-1-yl)methanone

6-Chloro-4-methylpicolinic acid (2.0 g, 11.70 mmol) was dissolved in dichloromethane (20 mL). N, N-diisopropylethylamine (3.0 g, 23.40 mmol), 2-(7-azobenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (4.4 g, 11.70 mmol) were added in sequence under ice bath conditions. Finally, 2-phenylpyrrolidine (1.7 g, 11.70 mmol) was added. After the addition, the mixture was reacted at room temperature for 4 hours. After TLC showed that the reaction was complete, 200 mL of water was added to the system, and the mixture was extracted with ethyl acetate three times. The organic phases were combined and backwashed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 2.8 g of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 301.1.

Step 2: Preparation of (6-hydroxy-4-methylpyridin-2-yl)(2-phenylpyrrolidin-1-yl)methanone

Weigh (6-chloro-4-methylpyridin-2-yl)(2-phenylpyrrolidin-1-yl)methanone (1.1 g, 3.67 mmol), boric acid (341.0 mg, 5.51 mmol), palladium acetate (41.0 mg, 0.18 mmol), 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'tri-isopropyl-1,1'biphenyl (222.5 mg, 0.46 mmol), cesium carbonate (2.4 g, 7.34 mmol), then seal the system for nitrogen protection, add solvent N-methylpyrrolidone (10 mL) to dissolve, then ventilate the system with nitrogen three times, then react at 90°C for 3 hours, and monitor the reaction of the raw materials by LCMS. The system is quenched with water, extracted with ethyl acetate, concentrated, and the crude product is separated by chromatography column (dichloromethane/methanol=20/1) to obtain 530 mg of the title compound as a purple-black solid.

MS (ESI) m/z (M+H) ⁺ = 283.1.

Step 3: Preparation of 4-methyl-6-(2-phenylpyrrolidine-1-carbonyl)-1-(prop-2-yn-1-yl)pyridin-2(1H)-one

Dissolve (6-hydroxy-4-methylpyridin-2-yl)(2-phenylpyrrolidin-1-yl)methanone (530.0 mg, 1.88 mmol) in toluene (10 mL) and water (0.5 mL), add potassium carbonate (518.9 mg, 3.76 mmol), lithium bromide (323.0 g, 3.76 mmol), tetrabutylammonium bromide (66.0 mg, 0.21 mmol) and propargyl bromide (224.0 mg, 1.88 mmol) in sequence at room temperature, and react at 80°C for 8 hours. After TLC shows that the reaction is complete, add 100 mL of water to the system, extract with ethyl acetate three times, combine the organic phases, backwash the organic phases once with a saturated sodium chloride solution, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The crude product is purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 180 mg of the title compound as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 321.1.

Preparation Example 17: Preparation of N-ethyl-N-(4-fluorophenyl)-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of N-ethyl-4-fluoroaniline

To a dry round-bottom flask, add 4-fluoroaniline (2.0 g, 18.00 mmol), acetaldehyde (1.19 g, 27 mmol), sodium cyanoborohydride (1.2 g, 19.8 mmol), and methanol (15 mL) in sequence, and then place the reaction solution at room temperature for 2 hours. LC-MS monitors the reaction, and a small amount of raw materials remain. Add water to quench the reaction, extract three times with ethyl acetate, combine the organic phases, backwash the organic phases once with a saturated sodium chloride solution, dry over anhydrous sodium sulfate, spin dry the solvent, and purify the crude product by column chromatography to obtain 1.5 g of the title product as a yellow oil.

MS (ESI) m/z (M+H) ⁺ =140.1.

Step 2: Preparation of N-ethyl-N-(4-fluorophenyl)-6-oxo-1,6-dihydropyridine-2-carboxamide

To the reaction tube, add 6-oxo-1,6-dihydropyridine-2-carboxylic acid (1.32g, 9.48mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.48g, 12.93mmol), and pyridine (10mL) in sequence; after stirring evenly, add N-ethyl-4-fluoroaniline (1.2g, 8.62mmol) dropwise, and then place the reaction system at room temperature for 6 hours. After the reaction is completed, add water to quench the reaction, extract with ethyl acetate three times, dry over anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by silica gel column to obtain 1.36g of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 261.1.

Step 3: Preparation of N-ethyl-N-(4-fluorophenyl)-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Into the reaction tube, N-ethyl-N-(4-fluorophenyl)-6-oxo-1,6-dihydropyridine-2-carboxamide (1.26g, 4.84mmol), potassium carbonate (2.01g, 14.52mmol), lithium bromide (0.84g, 9.68mmol), tetrabutylammonium bromide (0.17g, 0.53mmol), toluene (15mL), water (1mL) were added in sequence, and 3-bromopropyne (0.86g, 7.26mmol) was slowly added at room temperature after stirring evenly, and the reaction system was placed in an 80℃ oil bath for 2 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 0.59g of the title product.

MS (ESI) m/z (M+H) ⁺ = 299.1.

Preparation Example 18: Preparation of N-(4-fluorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Dissolve 6-oxo-1,6-dihydropyridine-2-carboxylic acid (1.0 g, 7.19 mmol) in pyridine (15 mL), add 4-fluoro-N-methylaniline (0.9 g, 7.19 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.0 g, 10.79 mmol) at room temperature, and react at room temperature for 8 hours. After TLC shows that the reaction is complete, concentrate the solvent, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (dichloromethane/methanol = 10/1 (V:V)) to obtain 1.6 g of the title compound as a light yellow oil.

MS (ESI) m/z (M+H) ⁺ = 247.2.

Step 2: Preparation of N-(4-fluorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Dissolve N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (1.5 g, 6.21 mmol) in N,N-dimethylformamide (20 mL), add 3-bromopropyne (1.5 g, 12.42 mmol) and potassium carbonate (1.7 g, 12.42 mmol) at room temperature, and react at 50 ° C for 1 hour. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (petroleum ether/ethyl acetate = 1/3 (V:V)) to obtain 1.5 g of the title compound as a brown oil.

MS (ESI) m/z (M+H) ⁺ = 285.2.

Preparation Example 19: Preparation of 6-(3-phenylmorpholine-4-carbonyl)-1-(prop-2-yn-1-yl)pyridin-2(1H)-one

Step 1: Preparation of 6-(3-phenylmorpholine-4-carbonyl)pyridin-2(1H)-one

Dissolve 6-oxo-1,6-dihydropyridine-2-carboxylic acid (450.0 mg, 3.24 mmol) in pyridine (10 mL), add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (619.0 mg, 3.24 mmol) and 3-phenylmorpholine (528.0 mg, 3.24 mmol) in turn, and react at room temperature for 4 hours. After TLC shows that the reaction is complete, add 200 mL of water to the system, extract three times with ethyl acetate, combine the organic phases and backwash once with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained crude product is purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 900 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 285.1.

Step 2: Preparation of 6-(3-phenylmorpholine-4-carbonyl)-1-(prop-2-yn-1-yl)pyridin-2(1H)-one

6-(3-phenylmorpholine-4-carbonyl)pyridin-2(1H)-one (550.0 mg, 1.94 mmol) was dissolved in toluene (10 mL) and water (0.5 mL). Potassium carbonate (535.0 mg, 3.88 mmol), lithium bromide (337.0 g, 3.88 mmol), tetrabutylammonium bromide (66.0 mg, 0.21 mmol), and propargyl bromide (230.0 mg, 1.94 mmol) were added in sequence at room temperature. After the addition, the mixture was reacted at 80°C for 8 hours. After TLC showed that the reaction was complete, 100 mL of water was added to the system, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, backwashed once with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography (petroleum ether/ethyl acetate = 1/1) to obtain 310 mg of the title compound as a yellow liquid.

MS (ESI) m/z (M+H) ⁺ = 323.1.

Preparation Example 20: Preparation of N-methyl-N-(naphthalen-2-yl)-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of tert-butyl N-(naphthalen-2-yl)carbamate

Into the reaction tube, add naphthalene-2-amine (2.0g, 13.97mmol), di-tert-butyl dicarbonate (4.57g, 20.96mmol), methanol (30mL) in sequence, stir evenly and place at 100℃ for 3 hours. After the reaction is completed, add water to quench the reaction, extract with ethyl acetate three times, dry over anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by silica gel column to obtain 3.0g of the title product as an orange-red solid.

MS (ESI) m/z (M+H-56) ⁺ =188.2.

Step 2: Preparation of tert-butyl N-methyl-N-(naphthalen-2-yl)carbamate

To the reaction tube, tert-butyl N-(naphthalene-2-yl)carbamate (3.0 g, 12.33 mmol) and tetrahydrofuran (20 mL) were added in sequence, followed by slow addition of sodium hydride (0.36 g, 14.80 mmol) under stirring, and after uniform stirring, iodomethane (2.10 g, 14.80 mmol) was added dropwise, and the reaction system was placed at room temperature for 2 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 3.0 g of the title product as an orange-red solid.

MS(ESI) m/z(M+H-56) ⁺ =202.1.

Step 3: Preparation of N-methylnaphthalene-2-amine

To the reaction tube, tert-butyl N-methyl-N-(naphthalene-2-yl)carbamate (3.0 g, 11.66 mmol), trifluoroacetic acid (7.65 g, 67.09 mmol), and dichloromethane (10 mL) were added in sequence, and the reaction was allowed to react at room temperature for 4 hours. After the reaction was completed, saturated sodium bicarbonate solution was added to quench the reaction, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 1.70 g of the title product as a red oil.

MS (ESI) m/z (M+H) ⁺ =158.1.

Step 4: Preparation of N-methyl-N-(naphthalen-2-yl)-6-oxo-1,6-dihydropyridine-2-carboxamide

Into the reaction tube, add 6-oxo-1,6-dihydropyridine-2-carboxylic acid (0.96g, 6.87mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.93g, 15.26mmol), pyridine (5mL), dichloromethane (5mL), stir evenly, add N-methylnaphthalene-2-amine (1.2g, 7.63mmol), and then place the reaction at room temperature for 4 hours. After the reaction is completed, add water to quench the reaction, extract with ethyl acetate three times, dry over anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by silica gel column to obtain 1.30g of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 279.2.

Step 5: Preparation of N-methyl-N-(naphthalen-2-yl)-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Into the reaction tube, N-methyl-N-(naphthalen-2-yl)-6-oxo-1,6-dihydropyridine-2-carboxamide (1 g, 3.59 mmol), potassium carbonate (1.49 g, 10.77 mmol), lithium bromide (0.62 g, 7.18 mmol), tetrabutylammonium bromide (0.12 g, 0.36 mmol), N,N-dimethylformamide (15 mL) were added in sequence. After stirring evenly, 3-bromopropyne (0.64 g, 5.38 mmol) was slowly added at room temperature, and the reaction system was placed in a 65°C oil bath for 2 hours. After the reaction was completed, water was added to quench the reaction, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 0.40 g of the title product as a yellow syrup.

MS (ESI) m/z (M+H) ⁺ = 317.2.

Preparation Example 21: Preparation of N-(4-fluorophenyl)-N-methyl-6-oxo-4-(trifluoromethyl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of N-(4-fluorophenyl)-N-methyl-4-(trifluoromethyl)pyridine-2-carboxamide

Weigh 4-(trifluoromethyl)pyridine-2-carboxylic acid (1 g, 5.23 mmol) and dissolve it in pyridine (20 mL). Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.2 g, 6.28 mmol) and 4-fluoro-N-methylaniline (0.79 g, 6.28 mmol). The system reacts at room temperature for 12 hours. After LC-MS shows that the reaction is complete, the system is concentrated, the acid is adjusted, and reverse phase purification is performed to obtain 1.38 g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 299.1.

Step 2: Preparation of 2-((4-fluorophenyl)(methyl)carbamoyl)-4-(trifluoromethyl)pyridine 1-oxide

Weigh N-(4-fluorophenyl)-N-methyl-4-(trifluoromethyl)pyridine-2-carboxamide (1.38 g, 4.63 mmol), dissolve in dichloromethane (20 mL), add m-chloroperbenzoic acid (3.2 g, 18.52 mmol) under ice bath. React at room temperature for 12 hours. LC-MS shows that the reaction is complete, the system saturated sodium bicarbonate solution is adjusted to alkaline, extracted with dichloromethane 3 times, the organic phase is concentrated, and purified by silica gel column to obtain 1.4 g of crude product of the title compound.

MS (ESI) m/z (M+H) ⁺ = 315.1.

Step 3: Preparation of 6-chloro-N-(4-fluorophenyl)-N-methyl-4-(trifluoromethyl)pyridine-2-carboxamide

Weigh 2-((4-fluorophenyl)(methyl)carbamoyl)-4-(trifluoromethyl)pyridine 1-oxide (1.4 g, 4.46 mmol), dissolve in acetonitrile (20 mL), add phosphorus oxychloride (4.07 mL, 44.6 mmol) under ice bath. Heat to 100°C and react for 12 hours. LC-MS shows that the reaction is complete and the system is reduced to dryness. The saturated sodium bicarbonate solution is reduced, extracted with ethyl acetate 3 times, the organic phase is concentrated, and purified by silica gel column to obtain 660 mg of the title compound as a light white solid.

MS (ESI) m/z (M+H) ⁺ = 333.1.

Step 4: Preparation of N-(4-fluorophenyl)-N-methyl-6-oxo-4-(trifluoromethyl)-1,6-dihydropyridine-2-carboxamide

Weigh 6-chloro-N-(4-fluorophenyl)-N-methyl-4-(trifluoromethyl)pyridine-2-carboxamide (300 mg, 0.9 mmol) and dissolve it in N-methylpyrrolidone (20 mL), add boric acid (83.47 mg, 1.35 mmol), palladium acetate (20.21 mg, 0.09 mmol), 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'tri-isopropyl-1,1'-biphenyl (87.24 mg, 0.18 mmol), cesium carbonate (586.48 mg, 1.8 mmol), replace with nitrogen 3 times, heat to 100°C and react for 3 hours. After LC-MS shows that the reaction is complete, filter, concentrate the filtrate, and purify on a silica gel column to obtain 70 mg of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 315.1.

Preparation Example A1: Preparation of tert-butyl N-(5-amino-6-iodo-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Step 1: Preparation of tert-butyl N-[(tert-butoxy)carbonyl]-N-(3-methyl-5-nitropyridin-2-yl)carbamate

Weigh 3-methyl-5-nitropyridine-2-amine (9.0 g, 58.8 mmol) and dissolve it in dichloromethane (100 mL). Add di-tert-butyl dicarbonate (12.8 g, 58.8 mmol) and 4-dimethylaminopyridine (7.2 g, 16.0 mmol) in an ice-water bath. After the addition, react at room temperature for 3 hours. After TLC shows that the reaction is complete, add the system dropwise into (500 mL) water in an ice-water bath, extract three times with ethyl acetate, combine the organic phases and backwash once with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The crude product is purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 20.0 g of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 354.1.

Step 2: Preparation of tert-butyl N-(5-amino-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Tert-butyl N-[(tert-butoxy)carbonyl]-N-(3-methyl-5-nitropyridin-2-yl)carbamate (20.0 g, 56.7 mmol) was dissolved in methanol (200 mL), and palladium carbon (4.0 g, 10% (W/W)) was slowly added in batches at 0°C. After the addition, the hydrogen was replaced three times, and then the reaction was carried out under a hydrogen atmosphere at room temperature for 10 hours. TLC showed that the raw material was completely consumed. The system was filtered and concentrated to obtain 18.0 g of an oily product, which was directly used in the next step reaction.

MS (ESI) m/z (M+H) ⁺ = 324.1.

Step 3: Preparation of tert-butyl N-(5-amino-6-iodo-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Dissolve tert-butyl N-(5-amino-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (18.0 g, 55.7 mmol) in N,N-dimethylformamide (100 mL), add N-iodosuccinimide (12.5 g, 55.7 mmol) at 0°C, and return to room temperature to react for 1 hour. After TLC shows that the reaction is complete, add (1 L) water to the system, extract with ethyl acetate five times, combine the organic phases and backwash once with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained crude product is purified by column chromatography (dichloromethane/methanol = 20/1 (V:V)) to obtain 15.0 g of the title compound as a solid.

MS (ESI) m/z (M+H) ⁺ = 450.1.

Preparation Example A2: Preparation of tert-butyl (tert-butoxycarbonyl)(6-iodo-3-methyl-5-(2,2,2-trifluoroacetylamino)pyridin-2-yl)carbamate

Dissolve tert-butyl (5-amino-6-iodo-3-methylpyridin-2-yl)(tert-butoxycarbonyl)carbamate (4.493 g, 10 mmol) in dichloromethane (100 mL), add trifluoroacetic anhydride (1.67 mL, 12 mmol) and triethylamine (2.78 mL, 20 mmol) at room temperature, and react at room temperature for 2 hours. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with dichloromethane several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography to obtain 5.18 g of the title product.

MS (ESI) m/z (M+H) ⁺ = 546.1.

Preparation Example A3: Preparation of N-(2-chlorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 6-chloro-N-(2-chlorophenyl)-N-methylpyridine-2-carboxamide

To the reaction tube, add 2-chloro-N-methylaniline (1.0g, 7.06mmol), 6-chloropyridine-2-carboxylic acid (1.0g, 6.35mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.03g, 10.59mmol), pyridine (5mL), and dichloromethane (5mL) in sequence; after stirring evenly, place the reaction system at room temperature for 4 hours. After the reaction is completed, add water to quench the reaction, extract with ethyl acetate three times, dry over anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by silica gel column to obtain 0.80g of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 281.2.

Step 2: Preparation of N-(2-chlorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Into the reaction tube, add 6-chloro-N-(2-chlorophenyl)-N-methylpyridine-2-carboxamide (720mg, 2.56mmol), boric acid (0.24g, 3.84mmol), palladium acetate (29mg, 0.13mmol), 2-di-tert-butylphosphine-2′,4′,6′-triisopropyl-3,6-dimethoxy-1,1′-biphenyl (160mg, 0.32mmol), cesium carbonate (1.67g, 5.12mmol); replace nitrogen three times, then add N-methylpyrrolidin-2-one (5mL), stir evenly and react at 80°C for 2 hours. After the reaction is complete, add water to quench the reaction, extract with ethyl acetate three times, dry with anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by silica gel column to obtain 600mg of the title product as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 263.1.

Step 3: Preparation of N-(2-chlorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Into the reaction tube, N-(2-chlorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (0.70g, 2.66mmol), potassium carbonate (1.10g, 7.98mmol), lithium bromide (0.46g, 5.32mmol), tetrabutylammonium bromide (0.086g, 0.27mmol), N,N-dimethylformamide (10mL) were added in sequence; after stirring evenly, 3-bromopropyne (0.32g, 2.66mmol) was slowly added at room temperature, and the reaction system was placed in a 60℃ oil bath for 2 hours. After the reaction was completed, water was added to quench the reaction, extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 0.35g of the title product as a brown oil.

MS (ESI) m/z (M+H) ⁺ = 301.2.

Preparation Example A4: Preparation of tert-butyl N-((tert-butoxy)carbonyl)-N-(3-chloro-6-iodo-5-(trifluoroacetylamino)pyridin-2-yl)carbamate

Step 1: Preparation of tert-butyl N-((tert-butoxy)carbonyl)-N-(3-chloro-5-nitropyridin-2-yl)carbamate

Dissolve 3-chloro-5-nitropyridine-2-amine (3.0 g, 17.29 mmol) in dichloromethane (40 mL), add di-tert-butyl dicarbonate (9.4 g, 43.22 mmol) and 4-dimethylaminopyridine (0.4 g, 3.46 mmol) at room temperature, and react at room temperature for 8 hours. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (petroleum ether/ethyl acetate = 5/1 (V:V)) to obtain 2.4 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 374.1.

Step 2: Preparation of tert-butyl N-(5-amino-3-chloropyridin-2-yl)-N-((tert-butoxy)carbonyl)carbamate

Dissolve tert-butyl N-((tert-butoxy)carbonyl)-N-(3-chloro-5-nitropyridin-2-yl)carbamate (2.0 g, 5.35 mmol) in ethanol (30 mL) and water (10 mL), add iron powder (0.6 g, 10.7 mmol) and ammonium chloride (0.8 g, 16.05 mmol) at room temperature, and react at 80°C for 3 hours. After TLC shows that the reaction is complete, filter and remove the solid while hot, and concentrate the solvent to obtain a crude product, which is purified by column chromatography (dichloromethane/methanol = 15/1 (V:V)) to obtain 1.5 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 344.2.

Step 3: Preparation of tert-butyl N-(5-amino-3-chloro-6-iodopyridin-2-yl)-N-((tert-butoxy)carbonyl)carbamate

Dissolve tert-butyl N-(5-amino-3-chloropyridin-2-yl)-N-((tert-butoxy)carbonyl)carbamate (1.5 g, 4.36 mmol) in N,N-dimethylformamide (10 mL), add N-iodosuccinimide (1.08 g, 4.80 mmol) and trifluoroacetic acid (0.025 g, 0.22 mmol) at room temperature, and react at room temperature for 5 hours. After TLC shows that the reaction is complete, add saturated brine to the reaction system and extract with ethyl acetate several times. The residue was dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (petroleum ether/ethyl acetate = 3/1 (V:V)) to obtain 1.0 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 470.1.

Step 4: Preparation of tert-butyl N-((tert-butoxy)carbonyl)-N-(3-chloro-6-iodo-5-(trifluoroacetylamino)pyridin-2-yl)carbamate

Dissolve tert-butyl N-(5-amino-3-chloro-6-iodopyridin-2-yl)-N-((tert-butoxy)carbonyl)carbamate (1.0 g, 2.13 mmol) in dichloromethane (10 mL), add triethylamine (0.4 g, 4.26 mmol) and trifluoroacetic anhydride (0.5 g, 2.34 mmol) at room temperature, and react at room temperature for 1 hour. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (petroleum ether/ethyl acetate = 2/1 (V:V)) to obtain 1.0 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 566.1.

Preparation Example A5: Preparation of N-(4-fluorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Dissolve 6-oxo-1,6-dihydropyridine-2-carboxylic acid (1.0 g, 7.19 mmol) in pyridine (15 mL), add 4-fluoro-N-methylaniline (0.9 g, 7.19 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.0 g, 10.79 mmol) at room temperature, and react at room temperature for 8 hours. After TLC shows that the reaction is complete, concentrate the solvent, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (dichloromethane/methanol = 10/1 (V:V)) to obtain 1.6 g of the title compound as a light yellow oil.

MS (ESI) m/z (M+H) ⁺ = 247.2.

Step 2: Preparation of N-(4-fluorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Dissolve N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (1.5 g, 6.21 mmol) in N,N-dimethylformamide (20 mL), add 3-bromopropyne (1.5 g, 12.42 mmol) and potassium carbonate (1.7 g, 12.42 mmol) at room temperature, and react at 50 ° C for 1 hour. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (petroleum ether/ethyl acetate = 1/3 (V:V)) to obtain 1.5 g of the title compound as a brown oil.

MS (ESI) m/z (M+H) ⁺ = 285.2.

Preparation Example A6: Preparation of (5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methanol

Step 1: Preparation of 3-bromo-N,N-bis[(4-methoxyphenyl)methyl]-6-methyl-5-nitropyridin-2-amine

3-Bromo-2-chloro-6-methyl-5-nitropyridine (12 g, 47.72 mmol) and bis[(4-methoxyphenyl)methyl]amine (14.74 g, 57.26 mmol) were dissolved in tetrahydrofuran (60 mL), sodium carbonate (6.07 g, 57.26 mmol) was added, and the mixture was reacted at 75°C for 16 hours. After LC-MS showed that the reaction was complete, the system was filtered and concentrated, and purified and separated by silica gel column to obtain 11.7 g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 472.1.

Step 2: Preparation of ethyl 3-(6-(bis[(4-methoxyphenyl)methyl]amino)-5-bromo-3-nitropyridin-2-yl)-2-oxopropanoate

Weigh 3-bromo-N,N-bis[(4-methoxyphenyl)methyl]-6-methyl-5-nitropyridin-2-amine (11.7 g, 24.77 mmol), dissolve it in diethyl oxalate (10.86 g, 74.31 mmol), add 1,8-diazabicycloundec-7-ene (4.53 g, 29.72 mmol) dropwise at 40°C, and react at 40°C for 12 hours. LC-MS showed that a small amount of starting material remained. Water was added to the system and the mixture was extracted three times with ethyl acetate. The organic phase was concentrated and purified on a silica gel column to obtain 13 g of the title compound as a pale white solid.

MS (ESI) m/z (M+H) ⁺ = 572.1.

Step 3: Preparation of ethyl 5-(bis(4-methoxybenzyl)amino)-6-bromo-1H-pyrrolo[3,2-b]pyridine-2-carboxylate

Weigh 3-(6-(bis[(4-methoxyphenyl)methyl]amino)-5-bromo-3-nitropyridin-2-yl)-2-oxopropanoic acid ethyl ester (13g, 22.71mmol), dissolve in acetic acid (150mL), add iron powder (5.07g, 90.84mmol), and react at room temperature for 12 hours. LC-MS shows that the reaction is complete, water is added to the system, and ethyl acetate is extracted three times, washed with saturated sodium bicarbonate solution, the organic phase is concentrated, and purified on a silica gel column to obtain 8.5g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 524.1.

Step 4: Preparation of ethyl 5-(bis(4-methoxybenzyl)amino)-6-methyl-1H-pyrrolo[3,2-b]pyridine-2-carboxylate

Weigh 5-(bis(4-methoxybenzyl)amino)-6-bromo-1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid ethyl ester (8.5 g, 16.21 mmol) and dissolve it in 1,4-dioxane (80 mL), then add methylboric acid (3.88 g, 64.84 mmol), dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (0.59 g, 0.81 mmol), potassium carbonate (8.96 g, 64.84 mmol) in sequence, replace the nitrogen three times, heat to 110°C and react for 12 hours. After LC-MS shows that the reaction is complete, filter, concentrate the filtrate, and purify on a silica gel column to obtain 6 g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 460.2.

Step 5: Preparation of ethyl 5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridine-2-carboxylate

Dissolve 5-(bis(4-methoxybenzyl)amino)-6-methyl-1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid ethyl ester (6g, 13.06mmol) in tetrahydrofuran (60mL), add sodium hydride (0.41g, 16.98mmol) in batches under ice-water bath, stir for 20 minutes, add 2-(trimethylsilyl)ethoxymethyl chloride (2.83g, 16.98mmol), and react at room temperature for 2 hours. After TLC shows that the reaction is complete, add water to quench, extract three times with ethyl acetate, combine the organic phases and backwash once with saturated brine, dry over anhydrous sodium sulfate, and purify on a silica gel column to obtain 7.5g of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 590.3.

Step 6: Preparation of (5-(bis[(4-methoxyphenyl)methyl]amino)-6-methyl-1-[(2-(trimethylsilyl)ethoxy)methyl]-1H-pyrrolo[3,2-b]pyridin-2-yl)methanol

Dissolve 5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid ethyl ester (7.5 g, 12.72 mmol) in tetrahydrofuran (100 mL), add lithium aluminum hydride (0.72 g, 19.08 mmol) in batches under ice-water bath, gradually warm to room temperature for 1 hour, and the reaction is complete by LC-MS. Add water (1 mL) under ice-water bath, stir for 5 minutes, add 15% sodium hydroxide solution (1 mL) dropwise, then slowly add water (3 mL), stir at room temperature for 0.5 hour, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and purify on silica gel column to obtain 6.8 g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 548.3.

Preparation Example A7: Preparation of 3-fluoro-N-(4-fluorophenyl)-6-hydroxy-N-methylpyridine-2-carboxamide

Step 1: Preparation of 6-chloro-3-fluoro-N-(4-fluorophenyl)-N-methylpyridine-2-carboxamide

Weigh 6-chloro-3-fluoropyridine-2-carboxylic acid (1 g, 5.7 mmol) and dissolve it in pyridine (20 mL). Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.31 g, 6.84 mmol) and 4-fluoro-N-methylaniline (0.86 g, 6.84 mmol). After the addition is complete, the system is reacted at room temperature for 12 hours. After LC-MS shows that the reaction is complete, the system is concentrated, the acid is adjusted, and reverse phase purification is performed to obtain 1.46 g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 283.1.

Step 2: Preparation of 3-fluoro-N-(4-fluorophenyl)-6-hydroxy-N-methylpyridine-2-carboxamide

Weigh 6-chloro-3-fluoro-N-(4-fluorophenyl)-N-methylpyridine-2-carboxamide (1.46 g, 5.17 mmol) and dissolve it in N-methylpyrrolidone (10 mL). Then add boric acid (0.48 g, 7.75 mmol), palladium acetate (0.12 g, 0.52 mmol), 2-(di-tert-butylphosphine)-3,6-dimethoxy-2',4',6'tri-isopropylamine (2-(di-tert-butylphosphine)-3,6-dimethoxy-2',4',6'tri-isopropylamine) in turn. Propyl-1,1'-biphenyl (0.5 g, 1.03 mmol), cesium carbonate (3.37 g, 10.34 mmol), nitrogen replacement 3 times, heating the reaction system to 80°C and reacting for 12 hours. After LC-MS showed that the reaction was complete, the mixture was filtered, the filtrate was concentrated, and the silica gel column was purified to obtain 1.32 g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 265.1.

Preparation Example A8: Preparation of 3-methyl-N-(4-fluorophenyl)-6-hydroxy-N-methylpyridine-2-carboxamide

The title compound was prepared using a similar preparation method to the above Preparation Example A7 using corresponding commercial reagents.

MS (ESI) m/z (M+H) ⁺ = 261.1.

Preparation Example A9: Preparation of N-(6-bromo-3-iodo-5-methylpyridin-2-yl)-2,2,2-trifluoroacetamide

Step 1: Preparation of 6-bromo-3-iodo-5-methylpyridin-2-amine

Dissolve 6-bromo-5-methylpyridin-2-amine (3.0 g, 16.04 mmol) in N,N-dimethylformamide (30 mL), add N-iodosuccinimide (3.9 g, 17.64 mmol) and trifluoroacetic acid (0.09 g, 0.80 mmol) at room temperature, and react at room temperature for 2 hours. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 5.0 g of the title compound as a brown solid.

MS (ESI) m/z (M+H) ⁺ = 312.9.

Step 2: Preparation of N-(6-bromo-3-iodo-5-methylpyridin-2-yl)-2,2,2-trifluoroacetamide

Dissolve 6-bromo-3-iodo-5-methylpyridin-2-amine (1.0 g, 3.20 mmol) in dichloromethane (10 mL), add trifluoroacetic anhydride (1.01 g, 4.80 mmol) and triethylamine (0.65 g, 6.4 mmol) at room temperature, and react at room temperature for 2 hours. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (petroleum ether/ethyl acetate = 2/1 (V:V)) to obtain 1.2 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 408.8.

Preparation Example A10: Preparation of tert-butyl N-(3-bromo-5-nitropyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Weigh 3-bromo-5-nitropyridine-2-amine (5.0 g, 23.0 mmol) and dissolve it in dichloromethane (100 mL). Add di-tert-butyl dicarbonate (10.0 g, 46.0 mmol) and 4-dimethylaminopyridine (2.8 g, 23.0 mmol) in turn under ice bath conditions. After addition, react at room temperature for 3 hours. After TLC shows that the reaction is complete, add the system dropwise into (500 mL) water under ice bath, extract three times with ethyl acetate, combine the organic phases and backwash once with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained crude product is purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 9.0 g of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 418.1.

Preparation Example A11: Preparation of tert-butyl N-[(tert-butoxy)carbonyl]-N-(3-cyclopropyl-5-nitropyridin-2-yl)carbamate

Weigh tert-butyl N-(3-bromo-5-nitropyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (2.0 g, 4.8 mmol), cyclopropylboronic acid (413.0 mg, 4.8 mmol), potassium carbonate (1.3 g, 9.6 mmol), dissolve in dioxane/water (10.0 mL/1.0 mL), add [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (350 mg, 0.5 mmol). The system is reacted at 90°C for 1 hour. TLC shows that the raw material is completely consumed. Water (10 mL) is added, and ethyl acetate is extracted. The organic phase is backwashed once with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and spin-dried. The crude product is separated by a chromatography column (PE/EA=4/1) to obtain 1.0 g of the title compound as a yellow oil.

MS (ESI) m/z (M+H) ⁺ = 380.2.

Preparation Example A12: Preparation of tert-butyl N-[(tert-butoxy)carbonyl]-N-(5-nitro-3-(prop-1-en-2-yl)pyridin-2-yl)carbamate

Weigh tert-butyl N-(3-bromo-5-nitropyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (2.0 g, 4.8 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (805.0 mg, 4.8 mmol), potassium carbonate (1.3 g, 9.6 mmol), dissolve in dioxane/water (10.0 mL/1.0 mL), add [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (350 mg, 0.5 mmol). The system is reacted at 90°C for 1 hour. TLC showed that the starting material was completely consumed, water (10 mL) was added, extracted with ethyl acetate, the organic phase was backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and spin-dried. The crude product was separated by chromatography column (PE/EA=4/1) to obtain 1.2 g of the title compound as a yellow oil.

MS (ESI) m/z (M+H) ⁺ = 380.2.

Preparation Examples A13-A15:

Using corresponding commercial reagents and a preparation method similar to that of the above-mentioned Preparation Example A4, Preparation Examples A13-A15 can be prepared as shown in the following table.

Preparation Example A16: Preparation of tert-butyl (5-amino-3-ethyl-6-iodopyridin-2-yl)(tert-butoxycarbonyl)carbamate

Step 1: Preparation of 3-ethyl-5-nitropyridin-2-amine

Under ice-water bath conditions, a concentrated nitric acid/concentrated sulfuric acid mixed solution (1.4mL/11mL) was slowly added dropwise to a concentrated sulfuric acid (11mL) solution of 3-ethylpyridin-2-amine (4.0g, 32.74mmol); after the addition, the reaction system was stirred at room temperature for 16 hours. After the reaction was completed, the reaction mixture was diluted with water (10mL), the pH was adjusted to 7-8 with a saturated sodium bicarbonate aqueous solution, and then extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the solvent was concentrated to obtain 3.5g of a crude product, which was directly used in the next step without purification.

MS (ESI) m/z (M+H) ⁺ = 168.1.

Step 2: Preparation of tert-butyl (tert-butoxycarbonyl)(3-ethyl-5-nitropyridin-2-yl)carbamate

Dissolve 3-ethyl-5-nitro-pyridin-2-amine (4.37 g, 26.14 mmol) in tetrahydrofuran (300 ml), add 4-dimethylaminopyridine (10.5 g, 85.95 mmol) and di-tert-butyl dicarbonate (13.5 g, 61.93 mmol) at room temperature, and react the reaction system at room temperature for 2 hours. Concentrate the reaction solution, and purify the crude product by silica gel chromatography to obtain 3.50 g of the title compound as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 368.4.

Step 3: Preparation of tert-butyl (5-amino-3-ethylpyridin-2-yl)(tert-butoxycarbonyl)carbamate

Dissolve tert-butyl (tert-butoxycarbonyl)(3-ethyl-5-nitropyridin-2-yl)carbamate (3.5 g, 9.53 mmol) in methanol (30 mL) and add palladium/carbon (300 mg, 2.47 mmol). After replacing hydrogen three times, the reaction system was reacted in a hydrogen atmosphere at room temperature for 2 hours. After the reaction was completed, the reaction system was cooled to room temperature, filtered through diatomaceous earth, and the filter cake was washed with methanol. The filtrate was concentrated to obtain 3.20 g of the title compound.

MS (ESI) m/z (M+H) ⁺ = 338.3.

Step 4: Preparation of tert-butyl (5-amino-3-ethyl-6-iodopyridin-2-yl)(tert-butoxycarbonyl)carbamate:

Dissolve tert-butyl (5-amino-3-ethylpyridin-2-yl)(tert-butoxycarbonyl)carbamate (3.20 g, 9.48 mmol) in N,N-dimethylformamide (15 mL), add N-iodosuccinimide (6.3 g, 28.00 mmol) at room temperature; after the addition is complete, the reaction system is reacted at room temperature for 16 hours. After the reaction is completed, the reaction solution is diluted with water and then extracted with ethyl acetate. Combine the organic phases, then wash with saturated brine and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is concentrated and separated and purified by chromatography to obtain 2.20 g of the title compound.

MS (ESI) m/z (M+H) ⁺ = 464.2.

Preparation Example A17: Preparation of 6-(6-fluoro-1,2,3,4-tetrahydroquinoline-1-carbonyl)-1-(prop-2-yn-1-yl)-1,2-dihydropyridin-2-one

Using corresponding commercial reagents and a preparation method similar to that of the above-mentioned Preparation Example A5, Preparation Example A17 can be obtained.

MS (ESI) m/z (M+H) ⁺ = 311.1.

Preparation Example A18: Preparation of 6-chloro-5-fluoro-2-iodopyridin-3-amine

Step 1: Preparation of tert-butyl N-(6-chloro-5-fluoropyridin-3-yl)carbamate

5-Bromo-2-chloro-3-fluoropyridine (4.0 g, 19.01 mmol) was dissolved in 1,4-dioxane (80 mL), and tert-butyl carbamate (2.6 g, 22.81 mmol), tris(dibenzylideneacetone)dipalladium (1.7 g, 1.90 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (2.2 g, 3.80 mmol) and cesium carbonate (12.4 g, 38.02 mmol) were added at room temperature, and the reaction was carried out at 90°C for 8 hours after replacing the inert gas. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, and the mixture was extracted several times with ethyl acetate and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (petroleum ether/ethyl acetate = 5/1 (V:V)) to obtain 2.0 g of the title compound as a light yellow oil.

(ESI) m/z (M+H) ⁺ = 247.7.

Step 2: Preparation of 6-chloro-5-fluoropyridin-3-amine

Dissolve tert-butyl N-(6-chloro-5-fluoropyridin-3-yl)carbamate (1.9 g, 7.70 mmol) in a solution of hydrochloric acid (0.6 g, 15.02 mmol) in 1,4-dioxane (2 mL). After addition, react at room temperature for 2 hours. After TLC shows that the reaction is complete, concentrate the solvent, add sodium bicarbonate to the reaction system to adjust the system to neutrality, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (dichloromethane/methanol = 10/1 (V:V)) to obtain 1.0 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ =147.5.

Step 3: Preparation of 6-chloro-5-fluoro-2-iodopyridin-3-amine

Dissolve 6-chloro-5-fluoropyridin-3-amine (1.0 g, 6.82 mmol) in N,N-dimethylformamide (10 mL), add N-iodosuccinimide (3.1 g, 13.64 mmol) at room temperature, and react at room temperature for 8 hours. After TLC shows that the reaction is terminated, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (dichloromethane/methanol = 15/1 (V:V)) to obtain 0.2 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 273.5.

Preparation Example A19: N-(6-chloro-5-fluoro-2-iodopyridin-3-yl)-2,2,2-trifluoroacetamide

Preparation Example A19 can be obtained by using a similar preparation method to the above Preparation Example A2 using corresponding commercial reagents.

MS (ESI) m/z (M+H) ⁺ = 369.0.

Preparation Example A20: Preparation of N-methyl-N-(5-methylpyridin-2-yl)-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of N,5-dimethylpyridin-2-amine

5-Methylpyridin-2-amine (2g, 18.49mmol), formaldehyde solution (0.83g, 27.73mmol), methanol (30mL), acetic acid (1mL) were added to the reaction bottle in sequence; after the addition was completed, the reaction was stirred at room temperature for 1 hour, sodium cyanoborohydride (3.49g, 55.47mmol) was slowly added, stirred evenly, and the reaction was reacted at room temperature for 4 hours. After the reaction was complete, water was added to quench the reaction, extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 0.54g of the title product as a light yellow liquid.

MS (ESI) m/z (M+H) ⁺ = 123.1.

Step 2: Preparation of 6-methoxy-N-methyl-N-(5-methylpyridin-2-yl)pyridine-2-carboxamide

Into the reaction tube, N,5-dimethylpyridine-2-amine (220mg, 1.80mmol), 6-methoxypyridine-2-carboxylic acid (0.28g, 1.8mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (0.52g, 2.7mmol), pyridine (4mL), dichloromethane (2mL) were added in sequence, and the reaction system was placed at room temperature for 4 hours after stirring. After the reaction was completed, water was added to quench the reaction, and ethyl acetate was extracted three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 0.30g of the title product as a yellow oil.

MS (ESI) m/z (M+H) ⁺ = 258.2.

Step 3: Preparation of N-methyl-N-(5-methylpyridin-2-yl)-6-oxo-1,6-dihydropyridine-2-carboxamide

To the reaction bottle, 6-methoxy-N-methyl-N-(5-methylpyridin-2-yl)pyridine-2-carboxamide (290 mg, 1.13 mmol), acetic acid (4 mL), and hydrobromic acid (2 mL) were added in sequence, and the reaction was placed at 80 ° C for 2 hours. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate solution, followed by extraction with ethyl acetate three times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The resulting mixture was purified by silica gel column to obtain 200 mg of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 244.2.

Step 4: Preparation of N-methyl-N-(5-methylpyridin-2-yl)-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Into the reaction tube, N-methyl-N-(5-methylpyridin-2-yl)-6-oxo-1,6-dihydropyridine-2-carboxamide (0.21g, 0.86mmol), potassium carbonate (0.36g, 2.58mmol), lithium bromide (0.15g, 1.72mmol), tetrabutylammonium bromide (0.03g, 0.09mmol), N,N-dimethylformamide (10mL) were added in sequence, and 3-bromopropyne (0.15g, 1.29mmol) was slowly added at room temperature after stirring evenly, and then the reaction system was placed in a 60°C oil bath for 2 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 0.15g of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 282.2.

Preparation Examples A21-A26:

Using corresponding commercial reagents and a preparation method similar to the above-mentioned Preparation Example A20, Preparation Examples A21-A26 can be prepared as shown in the following table.

Preparation Example A27: Preparation of 6-chloro-2-iodo-5-methoxypyridin-3-amine

Step 1: Preparation of tert-butyl N-(6-chloro-5-methoxypyridin-3-yl)carbamate

To the reaction tube, 5-bromo-2-chloro-3-methoxypyridine (1.5 g, 6.74 mmol), tert-butyl carbamate (1.18 g, 10.11 mmol), cesium carbonate (4.39 g, 13.48 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.29 g, 0.51 mmol), palladium acetate (0.23 g, 1.01 mmol), and 1,4-dioxane (20 mL) were added in sequence. After replacing nitrogen three times, the reaction was placed at 100 ° C for 3 hours. After the reaction was complete, the reaction was extracted with ethyl acetate three times, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated, and the obtained mixture was purified by silica gel column to obtain 1.30 g of the title product as a yellow solid.

MS(ESI) m/z(M-56+H) ⁺ =259.1.

Step 2: Preparation of 6-chloro-5-methoxypyridin-3-amine

To the reaction tube, tert-butyl N-(6-chloro-5-methoxypyridin-3-yl)carbamate (1.3 g, 5.04 mmol), trifluoroacetic acid (4 mL), and dichloromethane (4 mL) were added in sequence, and the reaction was allowed to react at room temperature for 3 hours. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate solution, extracted three times with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated, and the obtained mixture was purified by silica gel column to obtain 0.60 g of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ =159.1.

Step 3: Preparation of 6-chloro-2-iodo-5-methoxypyridin-3-amine

To the reaction tube, 6-chloro-5-methoxypyridin-3-amine (600 mg, 3.78 mmol), N-iodosuccinimide (0.94 g, 4.16 mmol) and N,N-dimethylformamide (5 mL) were added in sequence. After stirring evenly, the reaction was allowed to react at room temperature for 2 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The mixture was dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 833 mg of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 285.1.

Preparation Example A28: Preparation of N-(5-fluoro-6-methylpyridin-2-yl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 5-fluoro-N,6-dimethylpyridin-2-amine

5-Fluoro-6-methylpyridin-2-amine (10.0 g, 79.28 mmol), paraformaldehyde (8.1 g, 158.6 mmol), sodium methoxide (42.8 g, 792.8 mmol) and sodium borohydride (9.04 g, 237.9 mmol) were dissolved in methanol (120.0 mL), and the reaction solution was stirred at 40 ° C for 16 hours. LCMS showed that the reaction was complete, the reaction solution was diluted with water, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. After concentration, the title compound was separated and purified by column chromatography to obtain 7.2 g of an off-white solid.

MS (ESI) m/z (M+H) ⁺ =141.1.

Step 2: Preparation of N-(5-fluoro-6-methylpyridin-2-yl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

6-Oxo-1,6-dihydropyridine-2-carboxylic acid (7.5 g, 53.91 mmol), 5-fluoro-N,6-dimethylpyridine-2-amine (5.0 g, 35.94 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (20.49 g, 53.91 mmol) and N,N-diisopropylethylamine (11.59 g, 89.86 mmol) were dissolved in N,N-dimethylformamide (120.0 mL), and the reaction solution was stirred at room temperature for 16 hours. LCMS showed that the reaction was complete, the reaction solution was diluted with water, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. After concentration, the title compound was separated and purified by column chromatography to obtain 5.6 g of a brown solid.

MS (ESI) m/z (M+H) ⁺ = 262.1.

Step 3: Preparation of N-(5-fluoro-6-methylpyridin-2-yl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

N-(5-fluoro-6-methylpyridin-2-yl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (4.0 g, 15.31 mmol) and 3-bromopropyne (3.6 g, 30.62 mmol) were dissolved in acetonitrile (50.0 mL), followed by the addition of potassium carbonate (3.6 g, 26.02 mmol). After the addition was completed, the reaction system was stirred at 70 ° C for 6 hours. LCMS showed that the raw materials were basically reacted. The reaction solution was diluted with water, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. After concentration, the target compound 1.4 g was separated and purified by column chromatography to obtain a brown oily compound.

MS (ESI) m/z (M+H) ⁺ = 300.1.

Preparation Example A29: Preparation of 6-chloro-2-iodo-5-(trifluoromethyl)pyridin-3-amine

6-Chloro-5-(trifluoromethyl)pyridin-3-amine (1g, 5.09mmol) was dissolved in N,N-dimethylformamide (10mL), N-iodosuccinimide (1.37g, 6.11mmol) was added at room temperature, trifluoroacetic acid (0.19mL, 2.54mmol) was added dropwise, and the mixture was reacted at 80°C for 8 hours. After LC-MS showed that the reaction was complete, the system was concentrated and purified by silica gel column to obtain 1.44g of the title compound as an oil.

MS (ESI) m/z (M+H) ⁺ = 323.1.

Preparation Example A30: Preparation of 3,5-dibromo-6-methylpyrazine-2-amine

Dissolve 6-methylpyrazine-2-amine (2.0 g, 18.33 mmol) in dichloromethane (20 mL), add N-bromosuccinimide (7.2 g, 40.33 mmol) in batches at 0°C, and react at 0°C for 1 hour. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (petroleum ether/ethyl acetate = 5/1 (V:V)) to obtain 2.1 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 265.9.

Preparation Example A31: Preparation of N-(2-chloro-4-fluorophenyl)-6-hydroxy-N,3-dimethylpyridine-2-carboxamide

Step 1: Preparation of 6-chloro-N-(2-chloro-4-fluorophenyl)-3-methylpyridine-2-carboxamide

Weigh 6-chloro-3-methylpyridine-2-carboxylic acid (1 g, 5.83 mmol) and dissolve it in pyridine (20 mL), add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.34 g, 7 mmol) and 2-chloro-4-fluoroaniline (1.02 g, 7 mmol). The system reacts at room temperature for 12 hours. After LC-MS shows that the reaction is complete, the system is concentrated, the acid is adjusted, and reverse phase purification is performed to obtain 1.74 g of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 299.1.

Step 2: Preparation of 6-chloro-N-(2-chloro-4-fluorophenyl)-N,3-dimethylpyridine-2-carboxamide

Weigh 6-chloro-N-(2-chloro-4-fluorophenyl)-3-methylpyridine-2-carboxamide (1.82 g, 5.82 mmol) and dissolve it in tetrahydrofuran (30 mL). Add sodium hydride (0.35 g, 8.73 mmol) in batches under ice bath. After the addition, stir for 15 minutes, and add iodomethane (0.54 mL, 8.73 mmol) dropwise under ice bath. The system is gradually warmed to room temperature and stirred for 2 hours. After LC-MS shows that the reaction is complete, the system is quenched with ammonium chloride solution, extracted with ethyl acetate 3 times, and the organic phase is dried and concentrated to obtain 1.82 g of the title compound as an oil.

MS (ESI) m/z (M+H) ⁺ = 313.1.

Step 3: Preparation of N-(2-chloro-4-fluorophenyl)-6-hydroxy-N,3-dimethylpyridine-2-carboxamide

Weigh 6-chloro-N-(2-chloro-4-fluorophenyl)-N,3-dimethylpyridine-2-carboxamide (200 mg, 0.65 mmol) and dissolve it in N-methylpyrrolidone (5 mL), add boric acid (48.23 mg, 0.78 mmol), palladium acetate (14.59 mg, 0.065 mmol), 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'tri-isopropyl-1,1'-biphenyl (63.01 mg, 0.13 mmol), cesium carbonate (423.57 mg, 1.3 mmol), replace with nitrogen three times, heat to 80°C and react for 12 hours. LC-MS shows that there is little starting material, stop the reaction, filter the system, and purify by reverse phase to obtain 0.85 g of the title compound as an off-white solid.

MS (ESI) m/z (M+H) ⁺ = 295.1.

Preparation Example A32: Preparation of N-(2-chloro-4-fluoro-3-methylphenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Preparation Example A32 can be prepared by using corresponding commercial reagents and a similar method to the above Preparation Example A31.

MS (ESI) m/z (M+H) ⁺ = 295.1.

Preparation Example A33: Preparation of tert-butyl N-(5-amino-4-fluoro-6-iodo-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Step 1: Preparation of tert-butyl N-(4-fluoropyridin-2-yl)carbamate

To the reaction bottle, 4-fluoropyridin-2-amine (2 g, 17.84 mmol), di-tert-butyl dicarbonate (5.84 g, 26.76 mmol), 4-dimethylaminopyridine (3 mg, 0.018 mmol) and dichloromethane (50 mL) were added in sequence. After stirring evenly, the reaction was allowed to react at room temperature for 2 hours. After the reaction was complete, it was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The obtained mixture was purified by silica gel column to obtain 1.78 g of the title product as a white solid.

MS(ESI) m/z(M-56+H) ⁺ =157.0.

Step 2: Preparation of tert-butyl N-(4-fluoro-3-methylpyridin-2-yl)carbamate

Add tert-butyl N-(4-fluoropyridin-2-yl)carbamate (1.7 g, 8.01 mmol) to tetrahydrofuran (15 mL), then add N,N,N',N'-tetramethylethylenediamine (3.31 g, 19.22 mmol), protect with nitrogen, react at -78 degrees for 30 minutes, then drop n-butyl lithium (1.54 g, 24.03 mmol), then react at -78 degrees for 1 hour, then drop iodomethane (3.41 g, 24.03 mmol), continue to react for 1 hour, warm to room temperature, and react at room temperature for 2 hours. After the reaction is completed, the reaction system is added dropwise to saturated ammonium chloride, extracted with ethyl acetate, dried, and spin-dried with petroleum ether to obtain 1.70 g of the title product as a white solid.

MS(ESI) m/z(M-56+H) ⁺ =171.0.

Step 3: Preparation of 4-fluoro-3-methylpyridin-2-amine

To the reaction tube, tert-butyl N-(4-fluoro-3-methylpyridin-2-yl)carbamate (1.6 g, 7.08 mmol), trifluoroacetic acid (3 mL), and dichloromethane (2 mL) were added in sequence, and the reaction was allowed to react at room temperature for 3 hours. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate solution, extracted three times with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated, and the obtained mixture was purified by silica gel column to obtain 0.85 g of the title product as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 127.1.

Step 4: Preparation of 5-bromo-4-fluoro-3-methylpyridin-2-amine

To the reaction bottle, 4-fluoro-3-methylpyridin-2-amine (786 mg, 6.23 mmol) and tetrahydrofuran (10 mL) were added in sequence, and then N-bromosuccinimide (1.22 g, 6.85 mmol) was slowly added under an ice-water bath. After stirring evenly, the reaction was continued at this temperature for 2 hours. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 1.1 g of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 205.1.

Step 5: Preparation of tert-butyl N-(5-bromo-4-fluoro-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

5-Bromo-4-fluoro-3-methylpyridin-2-amine (800 mg, 3.90 mmol), 4-dimethylaminopyridine (0.048 g, 0.39 mmol), tetrahydrofuran (10 mL) were added to the reaction bottle in sequence, followed by di-tert-butyl dicarbonate (1.79 g, 8.19 mmol), stirred evenly, and reacted at room temperature for 2 hours. After the reaction was complete, water was added to quench the reaction, extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 875 mg of the title product as a yellow solid.

MS(ESI) m/z(M-156+H) ⁺ =251.1.

Step 6: Preparation of tert-butyl N-[(tert-butoxy)carbonyl]-N-(5-[(diphenylmethylene)amino]-4-fluoro-3-methylpyridin-2-yl)carbamate

To the reaction tube, tert-butyl N-(5-bromo-4-fluoro-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (870 mg, 2.15 mmol), benzophenone imine (0.47 g, 2.58 mmol), cesium carbonate (2.0 g, 6.15 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (149.28 mg, 0.26 mmol), palladium acetate (57.92 mg, 0.26 mmol), and 1,4-dioxane (10 mL) were added in sequence. After replacing nitrogen three times, the reaction was placed at 100 ° C for 3 hours. After the reaction was complete, it was extracted three times with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by silica gel column to obtain 950 mg of the title compound as a brown solid.

MS (ESI) m/z (M+H) ⁺ = 506.3.

Step 7: Preparation of tert-butyl N-(5-amino-4-fluoro-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

To the reaction bottle, tert-butyl N-[(tert-butoxy)carbonyl]-N-(5-[(diphenylmethylene)amino]-4-fluoro-3-methylpyridin-2-yl)carbamate (1.1 g, 2.18 mmol), sodium acetate (27.07 mg, 0.33 mmol), hydroxylamine hydrochloride (454.46 mg, 6.54 mmol) were added in sequence, followed by methanol (10 mL), stirred evenly, and reacted at room temperature for 2 hours. After the reaction was complete, water was added to quench the reaction, extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 540 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 342.2.

Step 8: Preparation of tert-butyl N-(5-amino-4-fluoro-6-iodo-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

To a reaction test tube, tert-butyl N-(5-amino-4-fluoro-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (540 mg, 1.58 mmol), N-iodosuccinimide (0.39 g, 1.74 mmol) and N,N-dimethylformamide (10 mL) were added in sequence. After stirring evenly, the reaction was allowed to react at room temperature for 2 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 560 mg of the title compound as a brown solid.

MS(ESI) m/z(M-156+H) ⁺ =312.1.

Preparation Example A34: Preparation of tert-butyl N-(5-amino-6-iodo-3,4-dimethylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Preparation Example A34 can be prepared using corresponding commercial reagents and a similar method to Steps 4-8 of Preparation Example A33 above.

MS(ESI) m/z(M-156+H) ⁺ =308.1.

Preparation Example A35: Preparation of 2,6-dibromo-4-chloro-5-methylpyridin-3-amine

Step 1: Preparation of 4-chloro-5-methylpyridin-3-amine

5-Bromo-4-chloropyridin-3-amine (400.0 mg, 1.93 mmol), 1,1'-bis(diphenylphosphino)ferrocenepalladium dichloride (282.4 mg, 0.39 mmol) and potassium carbonate (533.4 mg, 3.86 mmol) were weighed into a reaction tube, nitrogen was replaced, a solution of trimethylcyclotriboroxane (484.5 mg, 3.86 mmol) in 1,4-dioxane (5 mL) and water (1 mL) were added, and nitrogen was replaced again. The system was heated to 80°C and reacted for 8 hours. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, extracted with ethyl acetate several times, and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane/ethyl acetate = 2/1 (V:V)) to obtain 100.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 143.2.

Step 3: Preparation of 2,6-dibromo-4-chloro-5-methylpyridin-3-amine

Dissolve 4-chloro-5-methylpyridin-3-amine (80.0 mg, 0.56 mmol) in tetrahydrofuran (3 mL), add N-bromosuccinimide (219.2 mg, 1.23 mmol) under ice bath, and slowly return to room temperature to react for 2 hours. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is purified by column chromatography (dichloromethane/ethyl acetate = 1/1 (V:V)) to obtain 120.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 298.8.

Preparation Example A36: Preparation of 5-fluoro-N-(4-fluorophenyl)-6'-hydroxy-N-methyl-[3,4'-bipyridine]-2'-carboxamide

Step 1: Preparation of 4-bromo-N-(4-fluorophenyl)-N-methylpyridine-2-carboxamide

Weigh 4-bromopyridine-2-carboxylic acid (10 g, 49.5 mmol) and dissolve it in pyridine (100 mL). Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (13.9 g, 59.4 mmol) and 4-fluoro-N-methylaniline (7.43 g, 59.4 mmol). After the addition is complete, the system is reacted at room temperature for 12 hours. After LC-MS shows that the reaction is complete, the system is concentrated, the acid is adjusted, and reverse phase purification is performed to obtain 15.18 g of the oily title compound.

MS (ESI) m/z (M+H) ⁺ = 309.1.

Step 2: Preparation of 4-bromo-2-((4-fluorophenyl)(methyl)carbamoyl)pyridine 1-oxide

Weigh 4-bromo-N-(4-fluorophenyl)-N-methylpyridine-2-carboxamide (15 g, 48.52 mmol), dissolve in dichloromethane (100 mL), add m-chloroperbenzoic acid (12.56 g, 72.78 mmol) under ice bath. After the addition is complete, react at room temperature for 12 hours. LC-MS shows that the reaction is complete, the system saturated sodium bicarbonate solution is reduced, dichloromethane is extracted 3 times, the organic phase is concentrated, and the title compound 15.78 g is obtained by silica gel column purification.

MS (ESI) m/z (M+H) ⁺ = 325.1.

Step 3: Preparation of 5-fluoro-2'-((4-fluorophenyl)(methyl)carbamoyl)-[3,4'-bipyridine] 1'-oxide

4-Bromo-2-((4-fluorophenyl)(methyl)carbamoyl)pyridine 1-oxide (500 mg, 1.54 mmol) and 3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (343.5 mg, 1.54 mmol) were weighed and dissolved in 1,4-dioxane solution (20 mL) and water (2 mL); then dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (112.68 mg, 0.15 mmol) and potassium carbonate (425.69 mg, 3.08 mmol) were added, and nitrogen was replaced 3 times. The system was reacted at 80°C for 2 hours. LC-MS showed that the reaction was complete, the system was filtered through diatomaceous earth, the filtrate was concentrated to dryness, and purified by silica gel column to obtain 520 mg of the title compound as a light white solid.

MS (ESI) m/z (M+H) ⁺ = 342.1.

Step 4: Preparation of 6'-chloro-5-fluoro-N-(4-fluorophenyl)-N-methyl-[3,4'-bipyridine]-2'-carboxamide

Weigh 5-fluoro-2'-((4-fluorophenyl)(methyl)carbamoyl)-[3,4'-bipyridine]1'-oxide (520 mg, 1.52 mmol), dissolve in acetonitrile (5 mL), add phosphorus oxychloride (1.39 mL, 15.2 mmol) under ice bath. After the addition is complete, heat to 80°C and react for 12 hours. LC-MS shows that the reaction is complete, the system is concentrated, the sodium bicarbonate solution is reduced, and the dichloromethane is extracted three times. The organic phase is dried, concentrated, and purified and separated by silica gel column to obtain 382 mg of the title compound.

MS (ESI) m/z (M+H) ⁺ = 360.1.

Step 5: Preparation of 5-fluoro-N-(4-fluorophenyl)-6'-hydroxy-N-methyl-[3,4'-bipyridine]-2'-carboxamide

Weigh 6'-chloro-5-fluoro-N-(4-fluorophenyl)-N-methyl-[3,4'-bipyridine]-2'-carboxamide (382 mg, 1.06 mmol) and dissolve it in N-methylpyrrolidone (8 mL), add boric acid (98.31 mg, 1.59 mmol), palladium acetate (23.8 mg, 0.106 mmol), 2-(di-tert-butylphosphine)-3,6-dimethoxy-2',4',6'tri-isopropyl-1,1'-biphenyl (102.75 mg, 0.21 mmol), cesium carbonate (690.74 mg, 2.12 mmol), replace with nitrogen 3 times, heat the system to 90 ° C and react for 5 hours. LC-MS shows that there is little starting material, stop the reaction, filter the system, and purify by reverse phase to obtain 55 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 342.1.

Preparation Example A37: Preparation of N-(4-fluorophenyl)-6'-hydroxy-N-methyl-5-(trifluoromethyl)-[3,4'-bipyridine]-2'-carboxamide

This preparation example can be prepared using a similar preparation method to the above-mentioned preparation example A36 using corresponding commercial reagents.

MS (ESI) m/z (M+H) ⁺ = 392.2.

Preparation Example A38: N-(4-Fluorophenyl)-N-methyl-6-oxo-4-phenyl-1,6-dihydropyridine-2-carboxamide

This preparation example can be prepared using a similar preparation method to the above-mentioned preparation example A36 using corresponding commercial reagents.

MS (ESI) m/z (M+H) ⁺ = 323.2.

Preparation Example A39: Preparation of N-(4-fluorophenyl)-N-methyl-4-(1-methylpyrazol-4-yl)-6-oxo-1-prop-2-ynylpyridine-2-carboxamide

Step 1: Preparation of N-(4-fluorophenyl)-N-methyl-4-(1-methylpyrazol-4-yl)-1-oxidopyridin-1-ium-2-carboxamide

4-Bromo-2-((4-fluorophenyl)(methyl)carbamoyl)pyridine 1-oxide (2.0 g, 6.15 mmol) was dissolved in 1,4-dioxane (50.0 mL), and 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.92 g, 9.23 mmol), tetrakistriphenylphosphine palladium (0.5 g, After the addition was completed, the reaction system was stirred at 80°C for 6 hours under a nitrogen atmosphere. LCMS showed that the raw materials were basically reacted. The reaction solution was diluted with water, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. After concentration, the target compound 1.54 g was separated and purified by column chromatography to obtain a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 327.1.

Step 2: Preparation of N-(4-fluorophenyl)-N-methyl-4-(1-methylpyrazol-4-yl)-6-oxo-1H-pyridine-2-carboxamide

N-(4-fluorophenyl)-N-methyl-4-(1-methylpyrazol-4-yl)-1-oxidopyridin-1-ium-2-carboxamide (1.4 g, 4.29 mmol) was dissolved in ultra-dry tetrahydrofuran (50.0 mL), and then triethylamine (0.65 g, 6.44 mmol) was slowly added. The temperature was lowered to 0°C, and trifluoroacetic anhydride (1.35 g, 6.44 mmol) was added. After the addition was completed, the reaction system was stirred at 25°C for 16 hours. LCMS showed that the raw materials were basically reacted. The reaction solution was diluted with water, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. After the solution was concentrated, it was separated and purified by column chromatography to obtain 1.0 g of the title compound.

MS (ESI) m/z (M+H) ⁺ = 327.1.

Step 3: Preparation of N-(4-fluorophenyl)-N-methyl-4-(1-methylpyrazol-4-yl)-6-oxo-1-prop-2-ynylpyridine-2-carboxamide

N-(4-fluorophenyl)-N-methyl-4-(1-methylpyrazol-4-yl)-6-oxo-1H-pyridine-2-carboxamide (900 mg, 2.76 mmol), 3-bromopropyne (656.2 mg, 5.52 mmol), potassium carbonate (762.3 mg, 5.52 mmol) were dissolved in acetonitrile (50.0 mL), and the reaction solution was stirred at 60°C for 6 hours. LCMS showed that the reaction was complete, the reaction solution was diluted with water, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. After concentration, the title compound was separated and purified by column chromatography to obtain 480 mg.

MS (ESI) m/z (M+H) ⁺ = 365.1.

Preparation Example A40: Preparation of 5-fluoro-6-isopropyl-N-methylpyridin-2-amine

Step 1: Preparation of 6-bromo-5-fluoropyridine-2-carbonyl azide:

Dissolve 6-bromo-5-fluoropyridine-2-carboxylic acid (10.0 g, 45.46 mmol) in 1,4-dioxane (100.0 mL), add diphenylphosphoryl azide (16.6 g, 68.18 mmol) and triethylamine (6.9 g, 68.18 mmol), and stir the reaction solution at room temperature for 16 hours. LCMS shows that the reaction is complete, the reaction solution is diluted with water, the aqueous phase is extracted with ethyl acetate, the organic phases are combined, washed with saturated brine, and dried over anhydrous sodium sulfate. After concentration, 9.3 g of brown solid compound is obtained, which is directly used in the next step reaction.

MS(ESI)m/z(M+H) ⁺ =244.9/246.9.

Step 2: Preparation of tert-butyl (6-bromo-5-fluoropyridin-2-yl)carbamate

9.3 g of 6-bromo-5-fluoropyridine-2-carbonyl azide was dissolved in 1.4-dioxane (100.0 mL), the system was cooled to 0°C, tert-butyl carbamate (5.3 g, 45.5 mmol) was added, and the reaction solution was stirred at 70°C for 3 hours. LCMS showed that the reaction was complete, the reaction solution was diluted with water, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. After concentration, the title compound was separated and purified by column chromatography to obtain 7.5 g of a yellow solid.

MS(ESI)m/z(M+H) ⁺ =291.0/293.0.

Step 3: Preparation of tert-butyl (6-bromo-5-fluoropyridin-2-yl)(methyl)carbamate

Dissolve tert-butyl (6-bromo-5-fluoropyridin-2-yl)carbamate (3.0 g, 10.31 mmol) in N,N-dimethylformamide (50.0 mL), then cool to 0°C, and slowly add sodium hydride (742.0 mg, 30.92 mmol). After the addition is complete, stir the reaction system at 25°C for 0.5 hours. Cool the system to 0°C again, add iodomethane (2.19 g, 15.46 mmol), and stir the reaction system at 25°C for 3 hours after the addition is complete. LCMS shows that the raw material is basically reacted. Dilute the reaction solution with water, extract the aqueous phase with ethyl acetate, combine the organic phases, wash with saturated brine, and dry with anhydrous sodium sulfate. After concentration, separate and purify by column chromatography to obtain 2.5 g of the title compound as a brown oil.

MS(ESI)m/z(M-56+H) ⁺ =249.1/251.1.

Step 4: Preparation of tert-butyl (5-fluoro-6-(prop-1-en-2-yl)pyridin-2-yl)(methyl)carbamate

Dissolve tert-butyl (6-bromo-5-fluoropyridin-2-yl)(methyl)carbamate (2.0 g, 6.55 mmol) in 1,4-dioxane (50.0 mL)/water (5.0 mL), and then add 2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.65 g, 9.83 mmol), tetrakistriphenylphosphine palladium (533.5 mg, 0.65 mmol), and potassium phosphate (3.47 g, 16.39 mmol). After the addition is completed, the reaction system is stirred at 80 ° C for 16 hours. LCMS shows that the raw material is basically reacted. The reaction solution is diluted with water, the aqueous phase is extracted with ethyl acetate, the organic phases are combined, washed with saturated brine, and dried over anhydrous sodium sulfate. After concentration, it is separated and purified by column chromatography to obtain 1.5 g of the title compound as a brown oil.

MS (ESI) m/z (M+H) ⁺ = 267.1.

Step 5: Preparation of tert-butyl (5-fluoro-6-isopropylpyridin-2-yl)(methyl)carbamate

Dissolve tert-butyl (5-fluoro-6-(prop-1-en-2-yl)pyridin-2-yl)(methyl)carbamate (1.4 g, 5.26 mmol) in anhydrous methanol (50.0 mL), then slowly add palladium carbon (0.14 g). After the addition is complete, replace the hydrogen three times, and then place the reaction system under a hydrogen atmosphere at 25°C and stir for 16 hours. LCMS shows that the raw materials are basically reacted, and the reaction solution is filtered. After concentration, it is separated and purified by column chromatography to obtain 1.3 g of the title compound as a yellow oil.

MS (ESI) m/z (M+H) ⁺ = 269.1.

Step 6: Preparation of 5-fluoro-6-isopropyl-N-methylpyridin-2-amine

Dissolve tert-butyl (5-fluoro-6-isopropylpyridin-2-yl)(methyl)carbamate (1.3 g, 4.84 mmol) in dichloromethane (50.0 mL), then slowly add trifluoroacetic acid (10 mL). After the addition is complete, stir the reaction system at 25 ° C for 16 hours. LCMS shows that the raw materials are basically reacted. The reaction solution is adjusted to pH = 8 by adding sodium bicarbonate aqueous solution. The aqueous phase is extracted with dichloromethane, and the organic phases are combined, washed with saturated brine, and dried over anhydrous sodium sulfate. After the solution is concentrated, it is separated and purified by column chromatography to obtain 700 mg of the title compound as a yellow solid.

MS (ESI) m/z (M+H) ⁺ =169.1.

Preparation Example A41: Preparation of N-(5-fluoro-6-isopropylpyridin-2-yl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

This preparation example can be prepared using a similar preparation method to the above-mentioned preparation example A28 using corresponding commercial reagents.

MS (ESI) m/z (M+H) ⁺ = 328.1.

Preparation Example A42: Preparation of N-(5-fluoro-6-methylpyridin-2-yl)-N,3-dimethyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 4-fluoro-N,5-dimethylpyridin-2-amine

4-Fluoro-5-methylpyridin-2-amine (1g, 7.93mmol), paraformaldehyde (0.48g, 15.86mmol), sodium methoxide (2.57g, 47.58mmol), and methanol (10mL) were added to the reaction bottle in sequence, stirred evenly, and reacted at 40°C for 2 hours, then the reaction system was cooled to room temperature, sodium borohydride (1.20g, 31.72mmol) was slowly added, stirred evenly, and reacted at room temperature overnight. After the reaction was complete, water was added to quench the reaction, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 1.05g of the title product as a white solid.

MS (ESI) m/z (M+H) ⁺ = 141.2.

Step 2: Preparation of 6-chloro-N-(5-fluoro-6-methylpyridin-2-yl)-N,3-dimethylpyridine-2-carboxamide

To a dry reaction tube, add 5-fluoro-N,6-dimethylpyridin-2-amine (743 mg, 5.30 mmol), 6-chloro-3-methylpyridine-2-carboxylic acid (909.37 mg, 5.3 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.03 g, 10.6 mmol) and dichloromethane (10 mL); the reaction mixture was reacted at room temperature for 2 hours. After the reaction was complete, it was extracted with dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 1.3 g of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 294.1.

Step 3: Preparation of N-(5-fluoro-6-methylpyridin-2-yl)-N,3-dimethyl-6-oxo-1,6-dihydropyridine-2-carboxamide

6-Chloro-N-(5-fluoro-6-methylpyridin-2-yl)-N,3-dimethylpyridine-2-carboxamide (1.2 g, 4.09 mmol), 2-(di-tert-butylphosphine)-3,6-dimethoxy-2',4',6'tri-isopropyl-1,1'-biphenyl (0.25 g, 0.51 mmol), 2-(dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'tri-isopropyl-1,1'-biphenyl (0.25 g, 0.51 mmol) and 1,1'-dimethylpyridine (0.3 g, 0.51 mmol) were added to the reaction tube in sequence. Propyl-1,1′-biphenyl (0.38 g, 6.13 mmol), cesium carbonate (2.67 g, 8.18 mmol), palladium acetate (2.67 g, 8.18 mmol), replace nitrogen three times, add 1,4-dioxane (8 mL), stir evenly, react at 90°C overnight, after the reaction is complete, add water to quench the reaction, extract with ethyl acetate three times, dry over anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by silica gel column to obtain 887 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 276.1.

Step 4: Preparation of N-(5-fluoro-6-methylpyridin-2-yl)-N,3-dimethyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

N-(5-fluoro-4-methylpyridin-2-yl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (800 mg, 2.91 mmol), potassium carbonate (1.21 g, 8.73 mmol), tetrabutylammonium bromide (0.094 g, 0.29 mmol), lithium bromide (0.76 g, 8.73 mmol) were added to the reaction bottle in sequence, and 3-bromopropyne (0.52 g, 4.37 mmol) and N,N-dimethylformamide (8 mL) were slowly added after mixing evenly, and then the reaction was allowed to react at 80°C overnight. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 380 mg of the title product as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 314.2.

Preparation Example B1: Preparation of tert-butyl N-(5-amino-3-fluoro-6-iodo-2-pyridyl)-N-tert-butoxycarbonylcarbamate

Step 1: Preparation of 3-fluoro-5-nitropyridin-2-amine

Dissolve 2-chloro-3-fluoro-5-nitropyridine (5.0 g, 28.32 mmol) in ammonia methanol solution (50 mL, 7 M), and heat the reaction system to 50 ° C and stir for 16 hours. LCMS shows that the reaction is complete. After the reaction solution is concentrated, 4.17 g of yellow solid compound 3-fluoro-5-nitropyridine-2-amine is obtained, which is directly used in the next step reaction.

MS (ESI) m/z (M+H) ⁺ = 158.0.

Step 2: Preparation of tert-butyl N-(3-fluoro-5-nitro-2-pyridyl)-N-tert-butoxycarbonylcarbamate

Dissolve 3-fluoro-5-nitropyridine-2-amine (5.0 g, 38.21 mmol) in dichloromethane (40 mL), add di-tert-butyl dicarbonate (14.39 g, 76.43 mmol) and 4-dimethylaminopyridine (7.77 g, 76.43 mmol) in turn, and stir at room temperature for 16 hours. LCMS shows that the raw materials are basically reacted. Dilute the reaction solution with water, adjust the pH to 6 with dilute hydrochloric acid, extract with dichloromethane, wash the combined organic phases with saturated brine and dry with anhydrous sodium sulfate. After concentration, column chromatography separation and purification obtain 4.2 g of the title compound as a yellow solid.

MS (ESI) m/z (M+Na) ⁺ =380.2.

Step 3: Preparation of N-(5-amino-3-fluoro-2-pyridyl)-N-tert-butoxycarbonyl-carbamic acid tert-butyl ester

Dissolve tert-butyl N-(3-fluoro-5-nitro-2-pyridyl)-N-tert-butoxycarbonylcarbamate (4.2 g, 11.76 mmol) in a mixed solvent of methanol (40 mL) and water (8 mL), add ammonium chloride (3.24 g, 58.82 mmol) and iron powder (3.29 g, 58.82 mmol). After the addition is completed, place the reaction system The mixture was stirred at 60°C for 2 hours. LCMS showed that the reaction of the starting material was basically complete. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated and purified by column chromatography to obtain 3.6 g of the title compound as a brown solid.

MS (ESI) m/z (M+H) ⁺ = 328.2.

Step 4: Preparation of tert-butyl N-(5-amino-3-fluoro-6-iodo-2-pyridyl)-N-tert-butoxycarbonylcarbamate

Dissolve N-(5-amino-3-fluoro-2-pyridyl)-N-tert-butoxycarbonyl-carbamic acid tert-butyl ester (3.5 g, 10.69 mmol) in acetonitrile (40 mL), add N-iodosuccinimide (2.41 g, 10.69 mmol), and after the addition is completed, stir the reaction system at 25 ° C for 16 hours. LCMS shows that the raw materials are basically reacted. The reaction solution is quenched by adding saturated sodium sulfite solution, the aqueous phase is extracted with ethyl acetate, and the combined organic phases are washed with saturated brine and dried over anhydrous sodium sulfate. After concentration, it is separated and purified by column chromatography to obtain 4.2 g of the title compound as a brown solid.

MS (ESI) m/z (M+Na) ⁺ =476.3.

Preparation Example B2: Preparation of tert-butyl (tert-butyloxycarbonyl)(3-fluoro-6-iodo-5-(2,2,2-trifluoroacetylamino)pyridin-2-yl)carbamate

Preparation Example B2 can be obtained by using a similar preparation method to the above Preparation Example A2 using corresponding commercial reagents.

MS (ESI) m/z (M+H) ⁺ = 550.2.

Preparation Example B3: Preparation of N-[(4-fluorophenyl)methyl]-N-(1-methyl-1H-pyrazol-3-yl)-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of N-[(4-fluorophenyl)methyl]-1-methyl-1H-pyrazol-3-amine

4-Fluorobenzaldehyde (2g, 16.11mmol), 1-methyl-1H-pyrazole-3-amine (2.35g, 24.16mmol), and methanol (30mL) were added to the reaction bottle in sequence, and a catalytic amount of acetic acid was added dropwise and stirred evenly. The reaction was allowed to react at 40°C for 2 hours, and then the reaction system was cooled to room temperature, and sodium cyanoborohydride (0.029g, 0.76mmol) was slowly added. After stirring evenly, the reaction was allowed to react overnight at room temperature. After the reaction was complete, water was added to quench the reaction, and the reaction was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 1.5g of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 206.1.

Step 2: Preparation of N-[(4-fluorophenyl)methyl]-N-(1-methyl-1H-pyrazol-3-yl)-6-oxo-1,6-dihydropyridine-2-carboxamide

To a dry reaction tube, add N-[(4-fluorophenyl)methyl]-1-methyl-1H-pyrazole-3-amine (1.5 g, 7.31 mmol), 6-oxo-1,6-dihydropyridine-2-carboxylic acid (1.02 g, 7.31 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.80 g, 14.62 mmol), dichloromethane (10 mL), and pyridine (2 mL). The reaction mixture was reacted at room temperature for 1 hour. After the reaction was complete, it was extracted with dichloromethane. The organic layers were combined and dried over anhydrous sodium sulfate, filtered and rotary evaporated. The crude product was purified by silica gel column to obtain 1.3 g of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 327.2.

Step 3: Preparation of N-[(4-fluorophenyl)methyl]-N-(1-methyl-1H-pyrazol-3-yl)-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

N-[(4-fluorophenyl)methyl]-N-(1-methyl-1H-pyrazol-3-yl)-6-oxo-1,6-dihydropyridine-2-carboxamide (1.27 g, 3.89 mmol), potassium carbonate (1.61 g, 11.67 mmol), tetrabutylammonium bromide (0.13 g, 0.39 mmol), lithium bromide (1.01 g, 11.67 mmol), N,N-dimethylformamide (20 mL) were added to the reaction bottle in sequence, and 3-bromopropyne (0.69 g, 5.83 mmol) was slowly added after mixing evenly, and then the reaction was reacted at 80°C for 12 hours. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 0.90 g of the title product as a yellow liquid.

MS (ESI) m/z (M+H) ⁺ = 365.2.

Preparation Examples B4 to B9:

Using corresponding commercial reagents and a similar preparation method to the above-mentioned Preparation Examples A36 and A39, Preparation Examples B4 to B9 can be prepared as shown in the following table.

Preparation Examples B10 to B18:

Using the corresponding commercial reagents and the similar preparation methods as the above-mentioned Preparation Examples A3, A5, and A7, Preparation Examples B10 to B18 can be prepared as shown in the following table.

Preparation Example B19: Preparation of N-(4,5-difluoro-2-(1-methylcyclopropyloxy)phenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 1,2-difluoro-4-(1-methylcyclopropyloxy)-5-nitrobenzene

Weigh 1,2,4-trifluoro-5-nitrobenzene (1.0 g, 5.65 mmol) and dissolve it in N,N-dimethylformamide (10 mL). Add cesium carbonate (1.8 g, 5.65 mmol) and 1-methylcyclopropanol (406.0 mg, 5.65 mmol) in turn at room temperature. After the addition, react at 60°C for 6 hours. After TLC shows that the reaction is complete, add 200 mL of water to the system, extract with ethyl acetate three times, combine the organic phases and backwash once with saturated brine, dry with anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained crude product is separated and purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 900 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 230.1.

Step 2: Preparation of 4,5-difluoro-2-(1-methylcyclopropyloxy)aniline

1,2-Difluoro-4-(1-methylcyclopropyloxy)-5-nitrobenzene (900.0 mg, 3.93 mmol) was dissolved in methanol (50 mL), and palladium carbon (180.0 mg, 10% (W/W)) was slowly added in batches at 0°C. After the addition, the system was reacted at room temperature for 6 hours. LCMS showed that the starting material was completely consumed. The system was filtered and concentrated to obtain 500.0 mg of the title compound as a colorless oil.

MS (ESI) m/z (M+H) ⁺ = 200.2.

Step 3: Preparation of 6-chloro-N-(4,5-difluoro-2-(1-methylcyclopropyloxy)phenyl)pyridine-2-carboxamide

Dissolve 6-chloropyridine-2-carboxylic acid (394.0 mg, 2.51 mmol) in pyridine (5 mL), add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (480 mg, 2.51 mmol) and 4,5-difluoro-2-(1-methylcyclopropyloxy)aniline (500.0 mg, 2.51 mmol) in an ice bath, and react at room temperature overnight. After TLC shows that the reaction is complete, add 200 mL of water to the system, extract three times with ethyl acetate, combine the organic phases and backwash once with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained crude product is separated and purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 800 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 339.1.

Step 4: Preparation of 6-chloro-N-(4,5-difluoro-2-(1-methylcyclopropyloxy)phenyl)-N-methylpyridine-2-carboxamide

Weigh 6-chloro-N-(4,5-difluoro-2-(1-methylcyclopropyloxy)phenyl)pyridine-2-carboxamide (800mg, 2.37mmol) and dissolve it in N,N-dimethylformamide (10mL). Add sodium hydrogen sulfide (142.0mg, 3.56mmol) under ice bath, stir for 10min, add iodomethane (505mg, 3.56mmol) under this condition, and then naturally return to room temperature to react overnight. LCMS and TLC plates detected that the raw material reacted completely, the system was quenched with water, extracted with ethyl acetate, concentrated, and the crude product was separated by chromatography column (petroleum ether/ethyl acetate = 2/1) to obtain 600mg of the title compound as a black oil.

MS (ESI) m/z (M+H) ⁺ = 353.1.

Step 5: Preparation of N-(4,5-difluoro-2-(1-methylcyclopropyloxy)phenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Weigh 6-chloro-N-(4,5-difluoro-2-(1-methylcyclopropyloxy)phenyl)-N-methylpyridine-2-carboxamide (600.0 mg, 1.70 mmol), boric acid (158.0 mg, 2.56 mmol), palladium acetate (38.0 mg, 0.17 mmol), 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'tri-isopropyl-1,1'-biphenyl (165.0 mg, 0.34 mmol), and cesium carbonate (1.1 g, 3.40 mmol), then seal the system for nitrogen protection, add solvent 1,4-dioxane (10 mL) to dissolve, then ventilate the system with nitrogen three times, and then react at 90°C for 3 hours. LCMS monitors the complete reaction of the raw materials. LCMS and TLC plates showed that the raw material had reacted completely. The system was quenched with water, extracted with ethyl acetate, and concentrated. The resulting crude product was separated using a chromatography column (dichloromethane/methanol=20/1) to obtain 450 mg of the title compound.

MS (ESI) m/z (M+H) ⁺ = 335.1.

Step 6: Preparation of N-(4,5-difluoro-2-(1-methylcyclopropyloxy)phenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide

Weigh N-(4,5-difluoro-2-(1-methylcyclopropyloxy)phenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (450.0 mg, 1.35 mmol) and dissolve it in N,N-dimethylformamide (5 mL). Add cesium carbonate (438.0 mg, 1.35 mmol) and propargyl bromide (160.0 mg, 1.35 mmol) in turn at room temperature. After the addition, react at room temperature for 4 hours. After TLC shows that the reaction is complete, add 200 mL of water to the system, extract with ethyl acetate three times, combine the organic phases, backwash the organic phases with saturated sodium chloride solution once, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained crude product is separated and purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 200 mg of the title compound.

MS (ESI) m/z (M+H) ⁺ = 373.2.

Preparation Examples B20 to B24:

Using the corresponding commercial reagents and a similar preparation method to the above Preparation Example 19, Preparation Examples B20 to B24 can be prepared as shown in the following table.

Example 1: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Weigh 1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-oxo-1,6-dihydropyridine-2-carboxylic acid (60 mg, 0.090 mmol) and dissolve it in pyridine (10 mL). Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (25.88 mg, 0.14 mmol) and N-methylaniline (14.47 mg, 0.14 mmol). The system was reacted at room temperature for 12 hours. After LC-MS showed that the reaction was complete, water was added to the system, and dichloromethane was extracted 3 times. The organic phase was concentrated and purified by silica gel column to obtain 30 mg of the title compound as a white solid.

MS (ESI) m/z (M+H)+ = 758.3.

Step 2: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide (30 mg, 0.040 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (3 mL) was added dropwise under an ice-water bath, and the mixture was reacted at room temperature for 1.5 hours. After LC-MS showed that the reaction was complete, the system was concentrated, 7M ammonia methanol (5 mL) was added dropwise, and the mixture was heated to 50°C for 0.5 hours. LC-MS showed that the reaction was complete, the system was concentrated, the mixture was purified by reverse phase preparation, and lyophilized to obtain 3.63 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 388.1.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.41(s,1H),7.35(s,1H),7.20–7.02(m,4H),6.68(d,J＝7.3Hz,2H),6.34(d,J＝9.1Hz,1 H), 6.09 (s, 1H), 5.94 (d, J = 6.8Hz, 1H), 5.37 (s, 2H), 5.12 (s, 2H), 3.35 (s, 3H), 2.11 (s, 3H).

Example 2: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-isopropyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-isopropyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

N-isopropyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide (50.0 mg, 0.20 mmol), (5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methanol (109.5 mg, 0.20 mmol) and triphenylphosphine (10.4 mg, 0.040 mmol) were dissolved in tetrahydrofuran (5 mL), and diisopropyl azodicarboxylate (10.1 mg, 0.050 mmol) was added at room temperature. After the addition, the mixture was reacted at room temperature for 12 hours. After TLC showed that the reaction was complete, water was added to quench the reaction, and the mixture was extracted several times with dichloromethane and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane/methanol=10/1 (V:V)) to obtain 50.0 mg of the title compound as a light yellow solid.

(ESI) m/z (M+H) ⁺ = 787.0.

Step 2: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-isopropyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-isopropyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide (50.0 mg, 0.064 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1530.0 mg, 13.42 mmol) was added at room temperature. After the addition, the mixture was reacted at room temperature for 3 hours. After TLC showed that the reaction was complete, the mixture was concentrated under reduced pressure, and ammonia (119.2 mg, 7.00 mmol) in methanol (5 mL) was added. After the addition, the mixture was reacted at room temperature for 2 hours. After TLC showed that the reaction was complete, the mixture was concentrated under reduced pressure, separated and purified by preparative HPLC, and lyophilized to obtain 16.6 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 416.5.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.31(s,1H),7.36(s,1H),7.22–6.99(m,5H),6.62(d,J＝8.0Hz,1H),6.28(d,J＝9.1Hz,1H),6.06(s,1H), 6.03(d,J＝6.7Hz,1H),5.32(s,2H),5.13(s,2H),4.90(hept,J＝6.9Hz,1H),2.12(s,3H),1.29–0.89(m,6H).

Example 3: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-((5-chloropyridin-2-yl)methyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 1-[(5-(bis[(4-methoxyphenyl)methyl]amino)-6-methyl-1-[(2-(trimethylsilyl)ethoxy)methyl]-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl]-N-[(5-chloropyridin-2-yl)methyl]-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl-6-oxo-1,6-dihydropyridine-2-carboxylic acid (60 mg, 0.090 mmol) and 1-(5-chloropyridin-2-yl)-N-methylmethylamine (70.47 mg, 0.45 mmol) were weighed and dissolved in tetrahydrofuran (15 mL), and N-methylimidazole (74 mg, 0.9 mmol) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (252 mg, 0.9 mmol) were added in sequence. The system was reacted at 50°C for 0.5 hour. After LC-MS showed that the reaction was complete, water was added to the system, and ethyl acetate was extracted three times. The organic phase was concentrated and purified by silica gel column to obtain 52 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 807.3.

Step 2: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-((5-chloropyridin-2-yl)methyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

1-[(5-(bis[(4-methoxyphenyl)methyl]amino)-6-methyl-1-[(2-(trimethylsilyl)ethoxy)methyl]-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl]-N-[(5-chloropyridin-2-yl)methyl]-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (52 mg, 0.064 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (6 mL) was added dropwise under an ice-water bath, and the mixture was reacted at room temperature for 1.5 hours. After LC-MS showed that the reaction was complete, the system was concentrated, 7M ammonia methanol (10 mL) was added dropwise, and the mixture was heated to 60°C for 0.5 hours. LC-MS showed that the reaction was complete, the system was concentrated, purified by reverse phase preparation, and lyophilized to obtain 10.77 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 437.2.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.53–10.38(m,1H),8.66–8.49(m,1H),7.89–7.75(m,1H),7.58–7.33(m,1H),7.32–7.13(m,2H),6.61–6.48(m,1H),6.40–6.23(m ,1H),6.00–5.83(m,1H),5.62–5.45(m,1H),5.15–4.92(m,3H),4.66(s,1H),4.40–3.81(m,1H),2.88–2.62(m,3H),2.14–2.08(m,3H).

Example 4: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-(1,2,3,4-tetrahydroisoquinoline-2-carbonyl)pyridin-2(1H)-one

The title compound was prepared by using the corresponding commercial reagents and the product in the above preparation example as raw materials and using a preparation method similar to that in the above Example 3.

MS (ESI) m/z (M+H) ⁺ = 414.2.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.47–10.38(m,1H),7.56–7.40(m,1H),7.28–7.11(m,3H),7.08–6.91(m,2H),6.77–6.56(m,1H),6.39–6.22(m,1H),5.99–5.80(m,1H ),5.70–5.50(m,1H),5.11–4.87(m,4H),4.40–4.32(m,1H),3.97–3.50(m,1H),2.72–2.62(m,2H),2.37–2.26(m,1H),2.10–1.94(m,3H).

Example 5: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-(1,2,3,4-tetrahydroquinoline-1-carbonyl)pyridin-2(1H)-one

The title compound was prepared by using the corresponding commercial reagents and the product in the above preparation example as raw materials and using a preparation method similar to that in Example 1.

MS (ESI) m/z (M+H) ⁺ = 414.2.

^1H NMR(400MHz,DMSO-d6)δ10.48(s,1H),8.08–6.78(m,5H),6.75–6.31(m,2H) ,6.18–5.24(m,4H),5.10(s,2H),4.38–4.21(m,1H),2.79–2.62(m,2H),2.19 -2.04(m,4H),1.90–1.11(m,2H).

Example 6: Preparation of 1-((5-amino-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of tert-butyl (2-((6-(methyl(phenyl)carbamoyl)-2-oxopyridin-1(2H)-yl)methyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate

Tert-butyl (2-(hydroxymethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate (0.2g, 0.51mmol) and N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide (0.232g, 1.02mmol) were dissolved in tetrahydrofuran (20mL), triphenylphosphine (0.267g, 1.02mmol) was added under ice-water bath, and then diethyl azodicarboxylate (0.177g, 1.02mmol) was added dropwise, and stirred at this temperature for 12 hours. After LC-MS showed that the reaction was complete, water was added to quench, and ethyl acetate was extracted three times. The organic phases were combined and backwashed once with saturated brine, dried over anhydrous sodium sulfate, and purified on a silica gel column to obtain 0.15g of the title compound as an oil.

MS (ESI) m/z (M+H) ⁺ = 604.3.

Step 2: Preparation of 1-((5-amino-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Tert-butyl (2-((6-(methyl(phenyl)carbamoyl)-2-oxopyridin-1(2H)-yl)methyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate (150 mg, 0.25 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (6 mL) was added dropwise under an ice-water bath, and the mixture was reacted at room temperature for 1.5 hours. After LC-MS showed that the reaction was complete, the system was concentrated, 7M ammonia methanol (10 mL) was added dropwise, and the mixture was heated to 60°C for 0.5 hours. LC-MS showed that the reaction was complete, the system was concentrated, purified by reverse phase preparation, and lyophilized to obtain 50 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 374.2.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.75–10.31(m,1H),7.60–7.37(m,2H),7.29–7.02(m,4H),6.80–6.50(m ,2H),6.40–6.22(m,2H),6.10–5.94(m,2H),5.40–5.25(m,3H),3.36(s,3H).

Example 7: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-[2,2'-bipyridyl]-6(1H)-one

Step 1: Preparation of 1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-[2,2'-bipyridyl]-6(1H)-one

1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-bromopyridin-2(1H)-one (250.0 mg, 0.36 mmol) and 2-(tributylstannyl)pyridine (143.9 mg, 0.39 mmol) were dissolved in anhydrous toluene (5.0 mL), followed by the addition of tetrakis(triphenylphosphine)palladium (41.1 mg, 0.036 mmol). After the addition was completed, the reaction system was placed under nitrogen protection and heated to 110° C. and stirred for 16 hours. LCMS showed that the raw materials reacted completely. After the reaction solution was concentrated, it was separated and purified by column chromatography to obtain 250.0 mg of the brown colloid title compound for the next reaction.

MS (ESI) m/z (M+H) ⁺ = 702.4.

Step 2: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-[2,2'-bipyridyl]-6(1H)-one

1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-[2,2'-bipyridine]-6(1H)-one (150.0 mg, 0.21 mmol) was dissolved in trifluoroacetic acid (5.0 mL), and the reaction system was stirred at room temperature for 1 hour. LCMS showed that the raw material was completely reacted, and the reaction solution was concentrated to obtain a crude compound. The crude product was then dissolved in ammonia methanol solution (10.0 mL, 7 M), and the reaction solution was stirred at 60°C for 1 hour. LCMS showed that the raw material was completely reacted, and after concentration, 23.0 mg of the title compound was obtained as a light yellow solid by preparative separation.

MS (ESI) m/z (M+H) ⁺ = 332.1.

1H NMR (400MHz, DMSO-d6) δ10.37(s,1H),8.70(d,J＝4.7Hz,1H),7.85(td,J＝7.8,1.7Hz,1H),7.55–7.43(m,2H),7.43(d,J＝7.9Hz, 1H),7.22(s,1H),6.55(dd,J＝9.2,1.0Hz,1H),6.27(dd,J＝6.8,1.1Hz,1H),5.42(s,2H),5.26(s,1H),5.19(s,2H),2.03(s,3H).

Example 8: Preparation of 1-[(5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl]-6-(1-isopropylpyrazol-4-yl)pyridin-2-one

Step 1: Preparation of 1-[[5-[bis[(4-methoxyphenyl)methyl]amino]-6-methyl-1-(2-trimethylsilylethoxymethyl)pyrrolo[3,2-b]pyridin-2-yl]methyl]-6-(1-isopropylpyrazol-4-yl)pyridin-2-one

1-[[5-[Bis[(4-methoxyphenyl)methyl]amino]-6-methyl-1-(2-trimethylsilylethoxymethyl)pyrrolo[3,2-b]pyridin-2-yl]methyl]-6-bromo-pyridin-2-one (120.0 mg, 0.17 mmol), 1-isopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (40.6 mg, 0.17 mmol) and potassium phosphate (75.3 mg, 0.34 mmol) were dissolved in 1,4-dioxane (4.0 mL) and water (1.0 mL), and then [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (14.4 mg, 0.17 mmol) was added. After the addition was completed, the reaction system was placed under nitrogen protection and heated to 80°C and stirred for 16 hours. LCMS showed that the raw material was completely reacted. The reaction solution was concentrated and separated and purified by column chromatography to obtain 120.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 733.3.

Step 2: Preparation of 1-[(5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl]-6-(1-isopropylpyrazol-4-yl)pyridin-2-one

1-[[5-[Bis[(4-methoxyphenyl)methyl]amino]-6-methyl-1-(2-trimethylsilylethoxymethyl)pyrrolo[3,2-b]pyridin-2-yl]methyl]-6-(1-isopropylpyrazol-4-yl)pyridin-2-one (120.0 mg, 0.90 mmol) was dissolved in trifluoroacetic acid (3.0 mL), and the reaction system was stirred at room temperature for 1 hour. LCMS showed that the raw material was completely reacted, and the reaction solution was directly concentrated under reduced pressure to obtain a crude compound. The crude product was directly dissolved in ammonia methanol solution (2.0 mL, 7 M) and stirred at 60 ° C for 1 hour. LCMS showed that the raw material was completely reacted, and after concentration, 13.7 mg of the title compound was obtained as a light yellow solid by preparative separation.

MS(ESI)(M+H) ⁺ =m/z 363.1.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.49(s,1H),7.87(s,1H),7.49–7.44(m,2H),7.30(s,1H),6.45(dd,J＝9.1,1.3Hz,1H),6.27(dd,J＝6.9,1.4Hz ,1H),5.57–5.52(m,1H),5.25(s,2H),5.19(s,2H),4.48(hept,J＝6.6Hz,1H),2.10(s,3H),1.39(d,J＝6.7Hz,6H).

Example 9: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-((3,4-dihydroquinolin-1(2H)-yl)methyl)pyridin-2(1H)-one

Step 1: Preparation of tert-butyl N-(5-amino-3-methyl-6-(3-(2-oxo-6-[(1,2,3,4-tetrahydroquinolin-1-yl)methyl]-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)pyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Add 1-(prop-2-yn-1-yl)-6-[(1,2,3,4-tetrahydroquinolin-1-yl)methyl]-1,2-dihydropyridin-2-one (580.0 mg, 2.08 mmol), N-(5-amino-6-iodo-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamic acid tert-butyl ester (1.1 g, 2.50 mmol), tetrakistriphenylphosphine palladium (240.0 mg, 0.21 mmol), and cuprous iodide (48.0 mg, 0.25 mmol) to the reaction bottle in sequence. After the addition is completed, replace nitrogen three times, then add triethylamine (12 mL) and dichloromethane (5 mL). After stirring evenly, react at room temperature for 2 hours. After the reaction is complete, add water to quench the reaction, extract with ethyl acetate three times, dry over anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product was purified by silica gel column to obtain 700.0 mg of the title product as a brown solid.

MS (ESI) m/z (M+H) ⁺ = 600.4.

Step 2: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-((3,4-dihydroquinolin-1(2H)-yl)methyl)pyridin-2(1H)-one

To a dry reaction tube, tert-butyl N-(5-amino-3-methyl-6-(3-(2-oxo-6-[(1,2,3,4-tetrahydroquinolin-1-yl)methyl]-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)pyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (40.0 mg, 0.07 mmol) and potassium tert-butoxide (15.04 mg, 0.13 mmol) were added in sequence; nitrogen was replaced three times, tetrahydrofuran (1 mL) and N,N-dimethylformamide (0.3 mL) were added under nitrogen protection, and then the reaction mixture was added. The mixture should be placed in an oil bath at 80°C for 1 hour. After the reaction is completed, water is added to quench the reaction, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase concentrated. The crude product is purified by Pre-HPLC and freeze-dried to obtain 6.2 mg of the title product as a white solid.

MS (ESI) m/z (M+H) ⁺ = 400.2.

¹ H NMR (400 MHz, DMSO-d ₆ )δ10.50(s,1H),7.44–7.36(m,1H),7.31(s,1H),6.89(d,J＝7.2Hz,1H),6.75(t ,J＝7.6Hz,1H),6.46(t,J＝7.3Hz,1H),6.41(d,J＝9.0Hz,1H),6.12(d,J＝8.2Hz, 1H),6.00(d,J＝6.9Hz,1H),5.95(s,1H),5.35(s,2H),5.10(s,2H),4.55(s,2H) ,3.30–3.23(m,2H),2.71(t,J＝5.9Hz,2H),2.11(s,3H),1.90(p,J＝6.0Hz,2H).

The following examples can be prepared using corresponding commercial reagents and the products in the above-mentioned preparation examples as raw materials, using preparation methods similar to those in the above-mentioned examples.

The analytical data of the above described embodiment, including NMR and HPLC data, are as follows:

Example 25: Preparation of 1-[(5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl]-N-benzyl-N-[(5-fluoropyridin-2-yl)methyl]-6-oxo-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of tert-butyl N-(6-(3-(6-(benzyl[(5-fluoropyridin-2-yl)methyl]carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)-3-methyl-5-(trifluoroacetylamino)pyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Into the reaction tube, N-benzyl-N-[(5-fluoropyridin-2-yl)methyl]-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide (500 mg, 1.47 mmol), N-[(tert-butoxy)carbonyl]-N-(6-iodo-3-methyl-5-(trifluoroacetylamino)pyridin-2-yl)carbamic acid tert-butyl ester (0.88 g, 1.62 mmol), tetrakistriphenylphosphine palladium (170 mg, 0.15 mmol), cuprous iodide (34 mg, 0.18 mmol) were added in sequence. After replacing nitrogen three times, triethylamine (5 mL) and dichloromethane (5 mL) were added. After stirring evenly, the reaction was reacted at room temperature for 1 hour. After the reaction was complete, water was added to quench the reaction, and ethyl acetate was extracted three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain 520 mg of the title product.

MS (ESI) m/z (M+H) ⁺ = 793.3.

Step 2: Preparation of tert-butyl N-(2-[(6-(benzyl[(5-fluoropyridin-2-yl)methyl]carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl]-6-methyl-1H-pyrrolo[3,2-b]pyridin-5-yl)-N-[(tert-butoxy)carbonyl]carbamate

N-(6-(3-(6-(benzyl[(5-fluoropyridin-2-yl)methyl]carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)-3-methyl-5-(trifluoroacetylamino)pyridin-2-yl)-N-[(tert-butyloxy)carbonyl]carbamic acid tert-butyl ester (260 mg, 0.33 mmol), bistriphenylphosphine palladium dichloride (46 mg, 0.066 mmol), cuprous iodide (6.3 mg, 0.033 mmol) were added to the reaction tube, and nitrogen was replaced three times. Then triethylamine (5 mL) was added, and the reaction was stirred evenly and reacted at 120 degrees for 1 hour. After the reaction was complete, water was added to quench the reaction, and ethyl acetate was extracted three times, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was directly used in the next reaction without further purification.

MS (ESI) m/z (M+H) ⁺ = 697.3.

Step 3: Preparation of 1-[(5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl]-N-benzyl-N-[(5-fluoropyridin-2-yl)methyl]-6-oxo-1,6-dihydropyridine-2-carboxamide

To a dry reaction tube, add N-(2-[(6-(benzyl[(5-fluoropyridin-2-yl)methyl]carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl]-6-methyl-1H-pyrrolo[3,2-b]pyridin-5-yl)-N-[(tert-butyloxy)carbonyl]carbamic acid tert-butyl ester (130 mg, 0.19 mmol) and trifluoroacetic acid (3.93 g, 34.45 mmol), and place the mixture at room temperature to react. After the reaction is completed, add water to quench the reaction, extract with ethyl acetate three times, dry over anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by Pre-HPLC and freeze-dried to obtain 40 mg of the title product as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 497.3.

¹ H NMR (400 MHz, DMSO-d ₆ )δ10.42–10.31(m,1H),8.58–8.42(m,1H),7.71–7.55(m,1H),7.51–7.38(m,1 H),7.37–7.25(m,3H),7.25–7.18(m,2H),7.14–7.05(m,2H),6.57–6.49(m,1H ),6.50–6.37(m,1H),5.83–5.77(m,1H),5.41–5.26(m,1H),5.23–5.03(m,3H) ,4.98–4.75(m,1H),4.63–4.50(m,1H),4.44–4.15(m,2H),2.13–2.11(m,3H).

Example 27: 1-((5-amino-6-fluoro-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 1-(3-(3-amino-6-chloro-5-fluoropyridin-2-yl)prop-2-yn-1-yl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

6-Chloro-5-fluoro-2-iodopyridin-3-amine (120.0 mg, 0.44 mmol), N-methyl-6-oxo-N-phenyl-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide (117.2 mg, 0.44 mmol), tetrakis(triphenylphosphine)palladium (101.7 mg, 0.088 mmol), cuprous iodide (16.7 mg, 0.088 mmol) were weighed into a reaction tube, inert gas was replaced, triethylamine (89.0 mg, 0.88 mmol) and dichloromethane (5 mL) were injected to dissolve, inert gas was replaced again, and the reaction was carried out at room temperature for 1 hour after the addition was completed. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, extracted several times with dichloromethane, and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 411.9.

Step 2: Preparation of 1-(3-(6-chloro-5-fluoro-3-(trifluoroacetylamino)pyridin-2-yl)prop-2-yn-1-yl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

1-(3-(3-amino-6-chloro-5-fluoropyridin-2-yl)prop-2-yn-1-yl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide (170.0 mg, 0.41 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic anhydride (103.3 mg, 0.49 mmol) and triethylamine (82.9 mg, 0.82 mmol) were added at room temperature. After the addition, the mixture was reacted at room temperature for 2 hours. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, and the mixture was extracted several times with dichloromethane and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (petroleum ether/ethyl acetate = 1/1 (V:V)) to obtain 130.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 507.8.

Step 3: Preparation of 1-((5-chloro-6-fluoro-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

1-(3-(6-chloro-5-fluoro-3-(trifluoroacetylamino)pyridin-2-yl)prop-2-yn-1-yl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide (130.0 mg, 0.26 mmol) was dissolved in dichloromethane (3 mL), and bistriphenylphosphine palladium dichloride (36.5 mg, 0.052 mmol), cuprous iodide (9.9 mg, 0.052 mmol) and triethylamine (2.2 g, 21.64 mmol) were added at room temperature. After the addition, the inert gas was replaced, and the temperature was raised to 110°C for 1 hour. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, extracted with ethyl acetate several times, and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (petroleum ether/ethyl acetate = 1/2 (V:V)) to obtain 100.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 411.8.

Step 4: Preparation of 1-((5-chloro-6-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

1-((5-chloro-6-fluoro-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide (100.0 mg, 0.24 mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (19.2 mg, 0.48 mmol) was added at room temperature, and 2-(trimethylsilyl)ethoxymethyl chloride (44.0 mg, 0.26 mmol) was added dropwise at room temperature for 10 minutes. After the addition, the mixture was reacted at room temperature for 2 hours. After TLC showed that the reaction was complete, saturated aqueous ammonium chloride solution was added to the reaction system, and the mixture was extracted several times with ethyl acetate and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (petroleum ether/ethyl acetate = 2/1 (V:V)) to obtain 60.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 542.1.

Step 5: Preparation of tert-butyl N-(6-fluoro-2-((6-(methyl(phenyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate

1-((5-chloro-6-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide (60.0 mg, 0.11 mmol) was dissolved in tetrahydrofuran (5 mL), and tert-butyl carbamate (25.7 mg, 0.22 mmol), BrettPhos (11.8 mg, 0.022 mmol), BrettPhos-G3-Pd (19.9 mg, 0.022 mmol) and cesium carbonate (71.6 mg, 0.22 mmol) were added at room temperature. After the addition, the inert gas was replaced, and the temperature was raised to 90°C for 5 hours. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, extracted with ethyl acetate several times, and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (petroleum ether/ethyl acetate = 1/2 (V:V)) to obtain 40.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 622.7.

Step 6: Preparation of 1-((5-amino-6-fluoro-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-methyl-6-oxo-N-phenyl-1,6-dihydropyridine-2-carboxamide

Tert-butyl N-(6-fluoro-2-((6-(methyl(phenyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate (30.0 mg, 0.048 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (3.0 g, 26.84 mmol) was added at room temperature and stirred for 2 hours. After TLC showed that the reaction was complete, the solvent was concentrated to remove the solvent, and ammonia (0.36 g, 21.00 mmol) in methanol (3 mL) was added. After the addition was complete, the reaction was allowed to react at room temperature for 3 hours. After TLC showed that the reaction was complete, the solvent was concentrated to obtain a crude product, which was separated and purified by preparative HPLC to obtain 6.0 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 392.4.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.55(s,1H),7.51–7.41(m,1H),7.21–7.05(m,4H),6.74(d,J＝7.4Hz,2H),6.34(d,J =9.0Hz,1H),6.15(s,1H),5.96(d,J=6.6Hz,1H),5.55(s,2H),5.36(s,2H),3.37(s,3H).

Example 28: 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-4-methyl-6-(2-phenylpyrrolidine-1-carbonyl)pyridin-2(1H)-one

Step 1: Preparation of tert-butyl N-(5-amino-3-methyl-6-(3-(4-methyl-2-oxo-6-(2-phenylpyrrolidine-1-carbonyl)-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)pyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Weigh 4-methyl-6-(2-phenylpyrrolidine-1-carbonyl)-1-(prop-2-yn-1-yl)pyridin-2(1H)-one (180.0 mg, 0.56 mmol), tert-butyl N-(5-amino-6-iodo-3-methylpyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (252.0 mg, 0.56 mmol), tetrakistriphenylphosphine palladium (64.0 mg, 0.6 mmol), cuprous iodide (21.0 mg, 0.11 mmol), add solvent triethylamine (5 mL) and dichloromethane (2 mL), purify the system with nitrogen three times, and then react at room temperature for 3 hours. LCMS monitors that the reaction of the raw materials is complete. The system is quenched with water, extracted with ethyl acetate, concentrated, and the crude product is separated by chromatography (dichloromethane/methanol=20/1) to obtain 150 mg of the title compound as a purple-black solid.

MS (ESI) m/z (M+H) ⁺ = 642.3.

Step 2: Preparation of tert-butyl N-[(tert-butoxy)carbonyl]-N-(3-methyl-6-(3-(4-methyl-2-oxo-6-(2-phenylpyrrolidine-1-carbonyl)-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)-5-(trifluoroacetylamino)pyridin-2-yl)carbamate

Weigh tert-butyl N-(5-amino-3-methyl-6-(3-(4-methyl-2-oxo-6-(2-phenylpyrrolidine-1-carbonyl)-1,2-dihydropyrrolidine-1-yl)prop-1-yn-1-yl)pyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (150.0 mg, 0.23 mmol) and dissolve in dichloromethane (5 mL). Add triethylamine (95.7 μL, 0.728 g/mL, 0.69 mmol) and trifluoroacetic anhydride (32.0 μL, 1.511 g/mL, 0.23 mmol) in sequence at 0°C. After addition, return to room temperature and react overnight. After TLC shows that the reaction is complete, add 50 mL of water to the system, extract three times with ethyl acetate, combine the organic phases, backwash the organic phases once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained crude product was purified by column chromatography (petroleum ether/ethyl acetate=1/1 (V:V)) to obtain 150 mg of the title compound as a black solid.

MS (ESI) m/z (M+H) ⁺ = 738.3.

Step 3: Preparation of tert-butyl N-[(tert-butoxy)carbonyl]-N-(6-methyl-2-[(4-methyl-2-oxo-6-(2-phenylpyrrolidine-1-carbonyl)-1,2-dihydropyridin-1-yl)methyl]-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate

Weigh tert-butyl N-[(tert-butoxy)carbonyl]-N-(3-methyl-6-(3-(4-methyl-2-oxo-6-(2-phenylpyrrolidine-1-carbonyl)-1,2-dihydropyridine-1-yl)prop-1-yn-1-yl)-5-(trifluoroacetylamino)pyridin-2-yl)carbamate (150.0 mg, 0.23 mmol), add triethylamine (5 mL) and dichloromethane (1 mL), then add bistriphenylphosphine palladium dichloride (32.0 mg, 0.05 mmol) and cuprous iodide (19.0 mg, 0.10 mmol), purify the system with nitrogen three times, and then react at 120°C for 2 hours. LCMS monitors that the reaction of the raw materials is complete. The system is quenched with water, extracted with ethyl acetate, and concentrated. The crude product is separated by chromatography (dichloromethane/methanol=20/1) to obtain 100 mg of the title compound as a purple-black solid.

MS (ESI) m/z (M+H) ⁺ = 642.3.

Step 4: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-4-methyl-6-(2-phenylpyrrolidine-1-carbonyl)pyridin-2(1H)-one

Tert-butyl N-[(tert-butoxy)carbonyl]-N-(6-methyl-2-[(4-methyl-2-oxo-6-(2-phenylpyrrolidine-1-carbonyl)-1,2-dihydropyridin-1-yl)methyl]-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate (100.0 mg, 0.16 mmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (2 mL) was added successively at room temperature, and the system was reacted at room temperature for 3 hours after the addition. LCMS showed that the starting material was completely consumed, the system was concentrated, the crude product was purified by Pre-HPLC, and freeze-dried to obtain 50 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 442.1.

¹ H NMR(400MHz,Chloroform-d)δ9.71–9.18(m,1H),7.39–7.32(m,1H),7.29(s,1H),7.26–7.19(m,2H),6.91–6.80(m,1H),6.34–6.24(m,2H ),5.59–5.47(m,1H),5.33–5.15(m,3H),4.53(s,2H),4.10–3.83(m,2H),2.54–2.40(m,1H),2.23(s,3H),2.21(s,3H),2.17–1.65(m,4H).

The following examples can be prepared using corresponding commercial reagents and the products in the above-mentioned preparation examples as raw materials, using preparation methods similar to those in the above-mentioned examples.

The analytical data of the above described embodiment, including NMR and HPLC data, are as follows:

Example A1: Preparation of 1-[(5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl]-N-(2-chlorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of tert-butyl N-[(tert-butoxy)carbonyl]-N-(6-(3-(6-[(2-chlorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)-3-methyl-5-(trifluoroacetylamino)pyridin-2-yl)carbamate

Into the reaction tube, N-(2-chlorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide (60 mg, 0.20 mmol), N-[(tert-butoxy)carbonyl]-N-(6-iodo-3-methyl-5-(trifluoroacetylamino)pyridin-2-yl)carbamic acid tert-butyl ester (87 mg, 0.16 mmol), tetrakistriphenylphosphine palladium (23 mg, 0.02 mmol), and cuprous iodide (4 mg, 0.02 mmol) were added in sequence; after replacing nitrogen three times, triethylamine (1 mL) and Dichloromethane (0.5 mL) was added, stirred evenly, and the reaction was allowed to react at room temperature for 1 hour. After the reaction was complete, water was added to quench the reaction, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, filtered under reduced pressure, and the organic phase was concentrated. The crude product was purified by silica gel column to obtain about 60 mg of the title product, which can be directly used in the next step.

MS (ESI) m/z (M+H) ⁺ = 718.3.

Step 2: Preparation of tert-butyl N-[(tert-butoxy)carbonyl]-N-(2-[(6-[(2-chlorophenyl)(methyl)carbamoyl]-2-oxo-1,2-dihydropyridin-1-yl)methyl]-6-methyl-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate

To the reaction tube, add tert-butyl N-[(tert-butoxy)carbonyl]-N-(6-(3-(6-[(2-chlorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)-3-methyl-5-(trifluoroacetylamino)pyridin-2-yl)carbamate (60 mg, 0.084 mmol), tetrakistriphenylphosphine palladium (9.7 mg, 0.0084 mmol), cuprous iodide (1.6 mg, 0.0084 mmol), replace nitrogen three times, then add triethylamine (1 mL) and dichloromethane (0.5 mL), stir evenly, react at 120°C for 30 minutes, after the reaction is complete, add water to quench the reaction, extract three times with ethyl acetate, dry over anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by silica gel column to obtain 30 mg of the title compound as a brown solid.

MS (ESI) m/z (M+H) ⁺ = 622.3.

Step 3: Preparation of 1-[(5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl]-N-(2-chlorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

To a dry reaction tube, add tert-butyl N-[(tert-butoxy)carbonyl]-N-(2-[(6-[(2-chlorophenyl)(methyl)carbamoyl]-2-oxo-1,2-dihydropyridin-1-yl)methyl]-6-methyl-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate (30 mg, 0.048 mmol), trifluoroacetic acid (2 mL), and dichloromethane (2 mL), and the mixture is placed at room temperature to react for 1 hour. After the reaction is completed, add saturated sodium bicarbonate solution to quench the reaction, extract with ethyl acetate three times, dry over anhydrous sodium sulfate, filter under reduced pressure, and concentrate the organic phase. The crude product is purified by Pre-HPLC and lyophilized to obtain 9.7 mg of the title product as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 422.2.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.42(s,1H),7.62–7.55(m,1H),7.52(d,J＝8.0Hz,1H),7.47–7.39(m,1H),7.36(s,1H),7.24(t,J＝7.8Hz,1H),7.19(dd,J＝9.2,6.9Hz,1H),6 .37(d,J＝9.1Hz,1H),6.11(s,1H),5.96(d,J＝6.6Hz,1H),5.57(d,J＝14. 8Hz, 1H), 5.20 (d, J = 14.9Hz, 1H), 5.13 (s, 2H), 3.26 (s, 3H), 2.12 (s, 3H).

Example A2: Preparation of 1-((5-amino-6-chloro-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

The title compound was prepared by using the corresponding commercial reagents and the product in the above preparation example as raw materials and using a preparation method similar to that in the above Example A1.

MS (ESI) m/z (M+H) ⁺ = 426.1.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.62(s,1H),7.69(s,1H),7.22(dd,J＝9.2,6.8Hz,1H),6.97(t,J＝8.5Hz,2H),6.89(dd,J＝8.8,4.9Hz,2H) ,6.36(d,J＝9.2Hz,1H),6.13(d,J＝2.0Hz,1H),6.03(d,J＝6.7Hz,1H),5.63(s,2H),5.32(s,2H),3.33(s,3H).

Example A3: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-3-fluoro-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-3-fluoro-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

(5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methanol (200 mg, 0.37 mmol) and 3-fluoro-N-(4-fluorophenyl)-6-hydroxy-N-methylpyridine-2-carboxamide (146.65 mg, 0.55 mmol) were dissolved in tetrahydrofuran (10 mL), triphenylphosphine (145.57 mg, 0.55 mmol) was added under ice bath, diethyl azodicarboxylate (96.65 mg, 0.55 mmol) was slowly added dropwise, nitrogen was replaced three times, and the mixture was stirred at room temperature for 12 hours. After LC-MS showed that the reaction was complete, water was added to quench the mixture, and ethyl acetate was extracted three times. The organic phases were combined and backwashed once with saturated brine, dried over anhydrous sodium sulfate, and purified on a silica gel column to obtain 110 mg of the title compound as an oil.

MS (ESI) m/z (M+H) ⁺ = 794.4.

Step 2: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-3-fluoro-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

1-((5-(bis(4-methoxybenzyl)amino)-6-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-3-fluoro-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (110 mg, 0.14 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (6 mL) was added dropwise under ice bath, and the mixture was reacted at room temperature for 1.5 hours. LC-MS showed that the reaction was complete, the system was concentrated, 7M ammonia methanol (10 mL) was added dropwise, and the mixture was reacted at room temperature for 1.5 hours. LC-MS showed that the reaction was complete, the system was concentrated, the product was purified by reverse phase preparation, and lyophilized to obtain 24.09 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 424.1.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.41(s,1H),7.39–7.33(m,2H),7.31–7.19(m,1H),6.97(t,J＝8.7Hz,2H),6.86–6.79(m,1H),6.45(dd,J＝ 10.1,5.5Hz,1H),6.11(d,J＝2.0Hz,1H),5.36(d,J＝15.0Hz,1H),5.21–5.11(m,3H),3.36(s,3H),2.11(s,3H).

Example A4: Preparation of 1-((5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N,3-dimethyl-6-oxo-1,6-dihydropyridine-2-carboxamide

The title compound was prepared by using the corresponding commercial reagents and the product in the above preparation example as raw materials and using a preparation method similar to that in the above Example A3.

MS (ESI) m/z (M+H) ⁺ = 420.2.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.52–10.19(m,1H),7.53–7.26(m,3H),7.14–6.97(m,3H),6.63–6.20(m,1H),6.11–5 .95(m,1H),5.62–5.23(m,1H),5.16–4.90(m,3H),3.48–2.67(m,3H),2.15–1.77(m,6H).

Example A5: Preparation of 1-((6-amino-5-methyl-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of 1-(3-(6-bromo-5-methyl-2-(trifluoroacetylamino)pyridin-3-yl)prop-2-yn-1-yl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Weigh N-(6-bromo-3-iodo-5-methylpyridin-2-yl)-2,2,2-trifluoroacetamide (300.0 mg, 0.73 mmol), N-(4-fluorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide (207.5 mg, 0.73 mmol), tetrakis(triphenylphosphine)palladium (84.3 mg, 0.073 mmol), cuprous iodide (13.9 mg, 0.073 mmol) and triethylamine (147.7 mg, 1.46 mmol) in a reaction tube, replace nitrogen three times, add dichloromethane (5 mL) and replace nitrogen again, react at room temperature for 2 hours after addition. After TLC shows that the reaction is complete, add saturated brine to the reaction system, extract with dichloromethane several times, and dry over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane/methanol=15/1 (V:V)) to obtain 250.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 565.1.

Step 2: Preparation of 1-((6-bromo-5-methyl-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Weigh 1-(3-(6-bromo-5-methyl-2-(trifluoroacetylamino)pyridin-3-yl)prop-2-yn-1-yl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (200.0 mg, 0.35 mmol), cuprous iodide (6.6 mg, 0.035 mmol), bistriphenylphosphine palladium dichloride (24.5 mg, 0.035mmol) in a reaction tube, after replacing nitrogen, triethylamine (1460.0mg, 14.43mmol) and dichloromethane (1mL) were added, and nitrogen was replaced again. The system was heated to 120°C and reacted for 1 hour. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, extracted with ethyl acetate several times, and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane/methanol=10/1 (V:V)) to obtain 100.0mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 469.2.

Step 3: Preparation of tert-butyl 6-bromo-2-((6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-5-methyl-1H-pyrrolo[2,3-b]pyridine-1-carboxylate

1-((6-bromo-5-methyl-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (40.0 mg, 0.085 mmol) was dissolved in dichloromethane (2 mL), and di-tert-butyl dicarbonate (18.5 mg, 0.085 mmol), triethylamine (17.2 mg, 0.17 mmol) and 4-dimethylaminopyridine (2.0 mg, 0.017 mmol) were added at room temperature, and the mixture was reacted at room temperature for 2 hours. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, and the mixture was extracted several times with ethyl acetate and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane/methanol=15/1 (V:V)) to obtain 30.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 569.1.

Step 4: Preparation of tert-butyl 6-(((tert-butoxy)carbonyl)amino)-2-((6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-5-methyl-1H-pyrrolo[2,3-b]pyridine-1-carboxylate

6-Bromo-2-((6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-5-methyl-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (20.0 mg, 0.035 mmol), tert-butyl carbamate (8.2 mg, 0.070 mmol), BrettPhos-G ₃ -Pd (6.3 mg, 0.0070 mmol), BrettPhos (3.7 mg, 0.0070 mmol) and cesium carbonate (22.8 mg, 0.070 mmol) were weighed in a reaction tube in sequence, and nitrogen was replaced three times. After tetrahydrofuran (2 mL) was added and nitrogen was replaced again, the system was heated to 80°C and reacted for 3 hours. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, extracted several times with ethyl acetate, and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane/ethyl acetate = 1/1 (V:V)) to obtain 10.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 606.2.

Step 5: Preparation of 1-((6-amino-5-methyl-1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

6-(((tert-Butyloxy)carbonyl)amino)-2-((6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-5-methyl-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (10.0 mg, 0.017 mmol) was dissolved in dichloromethane (1 mL), and zinc bromide (19.1 mg, 0.085 mmol) was added at room temperature. After the addition, the mixture was reacted at room temperature for 8 hours. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, and the mixture was extracted several times with dichloromethane and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was separated and purified by preparative HPLC to obtain 1.45 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 406.2.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.36(s,1H),7.36(s,1H),7.20(dd,J＝9.2,6.8Hz,1H),7.07–6.95(m,4H),6.30(dd,J＝9.1,1.3Hz, 1H), 6.05 (d, J = 6.7Hz, 1H), 6.02 (d, J = 2.0Hz, 1H), 5.40 (s, 2H), 5.22 (s, 2H), 3.35 (s, 3H), 2.07 (s, 3H).

Example A6: Preparation of 1-((5-amino-6-bromo-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of tert-butyl N-(5-amino-3-bromo-6-(3-(6-[(4-fluorophenyl)(methyl)carbamoyl]-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)pyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Weigh tert-butyl N-(5-amino-3-bromo-6-iodopyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (200.0 mg, 0.4 mmol), N-(4-fluorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide (114.0 mg, 0.4 mmol), tetrakistriphenylphosphine palladium (46.0 mg, 0.04 mmol), cuprous iodide (15.0 mg, 0.08 mmol) in a dry reaction tube, add solvent triethylamine (5 mL) and dichloromethane (2 mL), purify the system with nitrogen three times, and then react at room temperature for 3 hours. LCMS monitors the reaction of the raw materials. The system is quenched with water, extracted with ethyl acetate, concentrated, and the obtained crude product is separated by chromatography column (dichloromethane/methanol=20/1) to obtain 210 mg of the title compound.

MS (ESI) m/z (M+H) ⁺ = 670.2.

Step 2: Preparation of tert-butyl N-(3-bromo-6-(3-(6-[(4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)-5-(trifluoroacetylamino)pyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate

Weigh tert-butyl N-(5-amino-3-bromo-6-(3-(6-[(4-fluorophenyl)(methyl)carbamoyl]-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)pyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (210.0 mg, 0.3 mmol) and dissolve in dichloromethane (5 mL). Add triethylamine (125.0 μL, 0.728 g/mL, 0.9 mmol) and trifluoroacetic anhydride (42.0 μL, 1.511 g/mL, 0.3 mmol) at 0°C. After addition, return the mixture to room temperature and react overnight. After TLC shows that the reaction is complete, add 10 mL of water to the system, extract with ethyl acetate three times, combine the organic phases, backwash the organic phases once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The obtained crude product was purified by column chromatography (petroleum ether/ethyl acetate=1/1 (V:V)) to obtain 170 mg of the title compound.

MS (ESI) m/z (M+H) ⁺ = 766.2.

Step 3: Preparation of tert-butyl N-(6-bromo-2-[(6-[(4-fluorophenyl)(methyl)carbamoyl]-2-oxo-1,2-dihydropyridin-1-yl)methyl]-1H-pyrrolo[3,2-b]pyridin-5-yl)-N-[(tert-butoxy)carbonyl]carbamate

Weigh tert-butyl N-(3-bromo-6-(3-(6-[(4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)-5-(trifluoroacetylamino)pyridin-2-yl)-N-[(tert-butoxy)carbonyl]carbamate (170.0 mg, 0.2 mmol) in a dry reaction tube, add triethylamine (5 mL) and dichloromethane (1 mL), then add bistriphenylphosphine palladium dichloride (15.0 mg, 0.02 mmol) and cuprous iodide (8.0 mg, 0.04 mmol). The system is purged with nitrogen three times, and then reacted at 120°C for 1 hour. LCMS monitors the reaction of the raw materials. The system is quenched with water, extracted with ethyl acetate, concentrated, and the obtained crude product is separated by chromatography column (dichloromethane/methanol=20/1) to obtain 45 mg of the title compound.

MS (ESI) m/z (M+H) ⁺ = 670.2.

Step 4: Preparation of 1-((5-amino-6-bromo-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Weigh tert-butyl N-(6-bromo-2-[(6-[(4-fluorophenyl)(methyl)carbamoyl]-2-oxo-1,2-dihydropyridin-1-yl)methyl]-1H-pyrrolo[3,2-b]pyridin-5-yl)-N-[(tert-butoxy)carbonyl]carbamate (45.0 mg, 0.067 mmol) and dissolve it in dichloromethane (2 mL). Add trifluoroacetic acid (2 mL) at room temperature. After the addition, the system is reacted at room temperature for 3 hours. LCMS shows that the starting material is completely consumed. The system is concentrated, and the crude product is purified by Pre-HPLC and freeze-dried to obtain 8.0 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 470.1.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.62(s,1H),7.84(s,1H),7.22(dd,J＝9.2,6.8Hz,1H),6.98(t,J＝8.5Hz,2H),6.95–6.85(m,2H), 6.36(dd,J＝9.2,1.3Hz,1H),6.12(s,1H),6.07–6.01(m,1H),5.55(s,2H),5.32(s,2H),3.33(s,3H).

Embodiment A7-A9:

The corresponding commercial reagents and the products in the above-mentioned preparation examples were used as raw materials, and a preparation method similar to that of the above-mentioned Example A6 was used to prepare the compounds of Examples A7-A9. The compound information is shown in the following table.

Example A10: Preparation of 1-((5-amino-6-isopropyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

1-((5-amino-6-(prop-1-en-2-yl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide (50 mg, 0.12 mmol) was dissolved in methanol (10 mL), and palladium carbon (8.0 mg, 10% (W/W)) was added at 0°C. After the addition, the hydrogen was replaced three times, and the reaction system was placed in a hydrogen atmosphere and reacted at room temperature for 3 hours. LCMS showed that the raw material was completely consumed, the system was concentrated, and the crude product was purified by Pre-HPLC and freeze-dried to obtain 3.5 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 434.2.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.27(s,1H),7.36(s,1H),7.13(dd,J＝9.2,6.8Hz,1H),6.82(t,J＝8.5Hz,2H),6.77–6.68(m,2H),6.29(d,J＝8.9Hz,1H), 6.01(d,J＝2.0Hz,1H),6.00–5.89(m,1H),5.27(s,2H),5.10(s,2H),3.26(s,3H),2.97–2.81(m,1H),1.11(d,J＝6.6Hz,6H).

Example A11: Preparation of 1-((5-amino-6-fluoro-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-(6-fluoro-1,2,3,4-tetrahydroquinoline-1-carbonyl)pyridin-2(1H)-one

The title compound was prepared by using the corresponding commercial reagents and the product in the above preparation example as raw materials and using a preparation method similar to the above Example A5.

MS (ESI) m/z (M+H) ⁺ = 436.2.

¹ H NMR (400MHz, DMSO-d ₆ )δ10.63(s,1H),8.13–6.94(m,4H),6.74–6.31(m,2H),6.16–5.49(m,5H),5 .41–5.04(m,1H),4.37–3.30(m,2H),3.12–2.65(m,2H),2.16–1.19(m,2H).

The following examples can be prepared using corresponding commercial reagents and the products in the above-mentioned preparation examples as raw materials, using preparation methods similar to those in the above-mentioned examples.

The analytical data of the above described embodiment, including NMR and HPLC data, are as follows:

Example A35: Preparation of 1-((1-acetyl-5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Step 1: Preparation of tert-butyl N-((tert-butoxy)carbonyl)-N-(6-(3-(6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)-3-methyl-5-(trifluoroacetylamino)pyridin-2-yl)carbamate

N-((tert-butoxy)carbonyl)-N-(6-iodo-3-methyl-5-(trifluoroacetylamino)pyridin-2-yl)carbamic acid tert-butyl ester (300.0 mg, 0.55 mmol), N-(4-fluorophenyl)-N-methyl-6-oxo-1-(prop-2-yn-1-yl)-1,6-dihydropyridine-2-carboxamide (156.4 mg, 0.55 mmol), tetrakistriphenylphosphine palladium (63.5 mg, 0.055 mmol) and cuprous iodide (10.5 mg, 0.055 mmol) were weighed into a reaction tube, and nitrogen was replaced three times. Triethylamine (111.3 mg, 1.10 mmol) and dichloromethane (5 mL) were added and nitrogen was replaced again. After addition, the reaction was allowed to react at room temperature for 1 hour. After TLC showed that the reaction was complete, saturated brine was added to the reaction system, extracted several times with ethyl acetate, and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane/ethyl acetate = 1/1 (V:V)) to obtain 350.0 mg of the title compound as a yellow-green solid.

MS (ESI) m/z (M+H) ⁺ = 702.2.

Step 2: Preparation of tert-butyl N-((tert-butoxy)carbonyl)-N-(2-((6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-6-methyl-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate

Weigh tert-butyl N-((tert-butoxy)carbonyl)-N-(6-(3-(6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)prop-1-yn-1-yl)-3-methyl-5-(trifluoroacetylamino)pyridin-2-yl)carbamate (350.0 mg, 0.50 mmol), bistriphenylphosphine palladium dichloride (70.1 mg, 0.10 mmol) and cuprous iodide (19.0 mg, 0.10 mmol) in a reaction tube, replace nitrogen three times, add triethylamine (1.5 g, 14.43 mmol) and dichloromethane (5 mL), replace nitrogen again, and heat the system to 120 ° C for 1 hour. After TLC shows that the reaction is complete, the reaction is reversed. Saturated brine was added to the system, extracted with ethyl acetate several times, and dried over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane/ethyl acetate = 1/1 (V:V)) to obtain 280.0 mg of the title compound as a yellow solid.

MS (ESI) m/z (M+H) ⁺ = 606.2.

Step 3: Preparation of tert-butyl N-(1-acetyl-2-((6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-6-methyl-1H-pyrrolo[3,2-b]pyridin-5-yl)-N-((tert-butoxy)carbonyl)carbamate

Dissolve tert-butyl N-((tert-butoxy)carbonyl)-N-(2-((6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-6-methyl-1H-pyrrolo[3,2-b]pyridin-5-yl)carbamate (40.0 mg, 0.066 mmol) in dichloromethane (5 mL), add triethylamine (13.4 mg, 0.13 mmol), 4-dimethylaminopyridine (1.6 mg, 0.013 mmol) and acetic anhydride (8.1 mg, 0.079 mmol) at room temperature, and react at room temperature for 8 hours. After TLC shows that the reaction is complete, concentrate the solvent, add saturated sodium chloride aqueous solution to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. The solvent was concentrated to obtain a crude product, which was purified by column chromatography (dichloromethane//ethyl acetate=2/1 (V:V)) to obtain 30.0 mg of the title compound as a light yellow solid.

MS (ESI) m/z (M+H) ⁺ = 648.2.

Step 4: Preparation of 1-((1-acetyl-5-amino-6-methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-N-(4-fluorophenyl)-N-methyl-6-oxo-1,6-dihydropyridine-2-carboxamide

Dissolve tert-butyl N-(1-acetyl-2-((6-((4-fluorophenyl)(methyl)carbamoyl)-2-oxo-1,2-dihydropyridin-1-yl)methyl)-6-methyl-1H-pyrrolo[3,2-b]pyridin-5-yl)-N-((tert-butoxy)carbonyl)carbamate (20.0 mg, 0.031 mmol) in dichloromethane (3 mL), add trifluoroacetic acid (1530.0 mg, 13.42 mmol) at room temperature, and react at room temperature for 2 hours. After TLC shows that the reaction is complete, concentrate the solvent, add saturated sodium bicarbonate aqueous solution to the reaction system, extract with ethyl acetate several times, and dry over anhydrous sodium sulfate. Concentrate the solvent to obtain a crude product, which is separated and purified by preparative HPLC to obtain 5.8 mg of the title compound as a white solid.

MS (ESI) m/z (M+H) ⁺ = 448.2.

¹ H NMR (400MHz, DMSO-d ₆ )δ7.80(s,1H),7.36–7.31(m,1H),7.29–7.24(m,2H),7.14(t,J＝8.5Hz,2H),6.63(s,1H),6.35(d,J＝ 9.2Hz, 1H), 6.21 (d, J = 6.8Hz, 1H), 5.59 (s, 2H), 5.40 (s, 2H), 3.29 (s, 3H), 2.79 (s, 3H), 2.17 (s, 3H).

The following examples can be prepared using corresponding commercial reagents and the products in the above-mentioned preparation examples as raw materials, using a similar preparation method to the above-mentioned Examples 9, A1, A5, and A6.

The analytical data of the above described embodiment, including NMR and HPLC data, are as follows:

Experimental Example 1: Enzyme Activity Test

1. Prepare Assay buffer. The ingredients are: 20mM Bicine (Sigma-Aldrich, Cat#B8660), 25mM NaCl (Sigma, Cat#S7653), 2mM DTT (Sigma, Cat#43815), 0.005% gelatin (Sigma, Cat#V900863), 0.01% Tween-20, 2.6μM MTA (Sigma, Cat#D5011), pH adjusted to 7.6.

2. Preparation of reference compound (MRTX9768) and test compound. The stock solution concentrations of MRTX9768 (MedChemExpress, HY-138684, CAS No.: 2629314-68-5) and the test compound were both 10 mM. Appropriate amount of stock solution was diluted in DMSO to the following concentrations: 99.010, 34.650, 12.380, 3.663, 1.282, 0.458, 0.136, 0.048, 0.017, 0.005 μM, for a total of 10 concentration points.

3. Prepare solutions containing both PRMT5 enzyme (BPS, Cat#51045), histone H4 peptide substrate (customized by Gill Biochemical, no external catalog number), and solutions containing only histone H4 peptide substrate. Prepare enzyme-substrate solutions in Assay buffer, with PRMT5 enzyme concentration of 7.6 nM and histone H4 concentration of 0.32 μM. Prepare substrate solutions in Assay buffer, with histone H4 concentration of 0.32 μM, without PRMT5.

4. Dilute SAM (New England Biolabs, Cat#B9003S) with Assay buffer to a 2.6 μM solution for later use.

5. Preparation of the test solution. The components of the test solution are: LANCE Ultra Europium-anti-methyl-Histone H4 Arginine 3 (perkinelmer, product number TRF0414-M) with a concentration of 4 nM, LANCE Ultra ULigh-Streptavidin (perkinelmer, product number TRF0102-M) with a concentration of The concentration is 53.3 nM, the content of LANCE Detection Buffer (perkinelmer, product number CR97-100) is 10% (volume percentage), and the rest is ultrapure water.

6. Add 100nL compound solution or blank DMSO (positive well) to the 384-well plate, add 5μL enzyme-substrate solution or substrate solution, centrifuge at 1000rpm for 30 seconds, and incubate at 25 degrees Celsius for 30 minutes. Then add 5μL SAM solution, centrifuge at 1000rpm for 30 seconds, and incubate at 25 degrees Celsius for 90 minutes. At this time, the components in the test wells are: 3.8nM PRMT5, 160nM histone H4, 1.3μM SAM, 1% DMSO (containing different concentrations of compounds at the same time); the positive control wells are divided into: 3.8nM PRMT5, 160nM histone H4, 1.3μM SAM, 1% DMSO (without compounds); the negative control wells are divided into: 160nM histone H4, 1.3μM SAM, 1% DMSO (without compounds).

7. After incubation, add 10 μL of detection solution to all wells, incubate for 60 minutes, and then detect in TR-FRET mode (320/340 nm excitation, 665/620 nm detection) and record the results.

8. Result processing: Inhibition% = (1-(well to be tested-negative well)/(positive well-negative well))×100.

The log value of the concentration was used as the X-axis, and the percentage inhibition rate (Inhibition%) was used as the Y-axis. The log (inhibitor) vs. response-Variable slope of the analysis software GraphPad Prism 8 was used to fit the dose-effect curve, thereby obtaining the IC ₅₀ value of each compound on the enzyme activity. The experimental results are shown in Table 1 below:

Table 1 IC ₅₀ values of the compounds of the present invention against PRMT5

Note: ND means not detected

Conclusion: The above experiments show that the compounds of the present invention have excellent PRMT5 inhibitory activity.

Experimental Example 2: OCI-Ly19 (MTAP null) cell proliferation inhibition experiment

Cell plating:

1) Take the cells out of the incubator and place them on the workbench. Mix them by gently pipetting and count them using CounterStar.

2) Dilute the cells to the required density with fresh complete medium. OCI-Ly19 (Nanjing Kebai, catalog number: CBP60621, MEM+10% FBS+1% P/S) was plated at a density of 1×10^4 cells/well, 100 μL/well.

3) Use an electric dispenser to draw 100 μL of cell suspension into a 96-well plate and add 200 μL 1× PBS to the edge wells to prevent evaporation.

Formulated compound:

1) First, the stock solution of the compound was diluted from 10 mM to 2000, 600, 200, 60, 20, 6, 2, and 0.6 μM, with a DMSO content of 100%.

2) After thorough mixing, an electric dispenser was used to take out 2.6 μL of the above graded diluted compounds and DMSO (as a control) and added to 257.4 μL of culture medium. At this time, the mixture was diluted 100 times and the DMSO content was 1%.

3) After thoroughly mixing with a spray gun, take out the cells, set up two replicate wells for each concentration of each compound, add 100 μL of the compound diluted in step 2 to each of the two replicate wells, and make the final concentration of the compound: 10, 3, 1, 0.3, 0.1, 0.03, 0.01, 0.003, 0 μM, at this time, there are 200 μL of culture medium in each well of the cell culture plate, and the DMSO content is 0.5%.

4) The cells were returned to the 37°C, 5% CO ₂ incubator for continued culture and tested 5 days after compound treatment.

CTG test:

1) After culturing for a specific period of time, remove the cells and aspirate a portion of the culture medium to leave 50 μL of culture medium in each well. Use a dispenser to add 50 μL/well of CTG reagent (cell counting-Lite 2.0, Vazyme, DD1101-02).

2) Incubate on a shaker at room temperature for 15 minutes and allow to equilibrate at room temperature for 15 minutes.

3) Detection using a multifunctional microplate reader.

Data Analysis:

1) Calculate the cell viability, Cell viability % = As/Ac × 100%. As: test well detection value (culture medium containing cells, CTG, test compound), Ac: control well detection value (culture medium containing cells, CTG, no test compound).

2) Using the log value of the compound concentration as the X-axis and the cell viability (Cell viability %) as the Y-axis, a dose-effect curve was fitted to obtain the IC ₅₀ value of each compound's cell proliferation inhibitory activity.

Table 2 Proliferation inhibition activity of the compounds of the present invention on OCI-Ly19 cells

The test results show that the compounds of the present application have significant proliferation inhibition activity on MTAP-deficient cells. The inhibitory activity of the compounds of the present application on OCI-Ly19 cells is more than 10 to 20 times that of the prior art compounds; the inhibitory activity of some compounds of the present application on OCI-Ly19 cells is 20 to 50 times that of the prior art compounds; and the inhibitory activity of some compounds of the present application on OCI-Ly19 cells is 50 to 100 times that of the prior art compounds.

Experimental Example 3: Liver microsome metabolic stability test

Liver microsome source information:

The compound to be tested was prepared to 10mM with DMSO, 2μL was added to 198μL 50% acetonitrile/50% aqueous solution to obtain a 100μM solution. Liver microsomes were taken and prepared to 0.5mg/mL with PBS, NADPH cofactor (final concentration of 1mM) was added, and the mixture was preheated at 37°C for 10 minutes. 2.5μL of the prepared compound to be tested was taken, and 222.5μL of the above preheated mixture was added and placed in a 37°C water bath to start the reaction. At 0.5, 15, 30 and 45 minutes, the incubated centrifuge tubes were taken out and 25μL of the incubated liver microsome suspension (containing the compound) was taken out, and 5 times the volume of the stop solution was added to stop the reaction, and the supernatant was centrifuged at 3220g for 40 minutes. After taking the supernatant, the remaining compound content in the sample at each time point was detected by LC-MS/MS. After nonlinear linear regression of the % remaining drug amount-time, the half-life (t _1/2 ) of the compound in liver microsomes was calculated. Exemplary experimental results are shown in Table 3.

Table 3. Half-life (t _1/2 ) of the compounds of the present invention in liver microsomes

The test results show that the compounds of the present application show good in vitro metabolic stability in in vitro human, rat and mouse liver microsome stability tests (for example, the in vitro rat and mouse liver microsome metabolism t _1/2 are both greater than 30 min, and the in vitro human liver microsome metabolism t _1/2 is greater than 60 min).

Experimental Example 4: HCT116 WT cells (Source: Nanjing Kebai) activity test

On the first day, cell plating: after trypsinization, cells were resuspended to the required density with complete medium (McCoy5A containing 10% FBS. FBS brand is ExCell Bio, catalog number FND500; McCoy5A brand is BOSTER, catalog number PYG0025), mixed evenly, and added to a 96-well plate at 100 μL/well, with a cell density of 500 to 1000 cells per well, and returned to the incubator for cell attachment and growth. On the second day, test compounds were added: cells were starved with serum-free medium for 4 hours before adding compounds, and then complete medium containing corresponding concentrations of compounds was added, and cultured at 37°C, 5% CO ₂ for 120 hours. On the 7th day, the cells treated with the compound were taken out and equilibrated to room temperature, and a portion of the culture medium in the wells of the culture plate was aspirated to leave 50 μL of culture medium in each well, and then 50 μL of CTG reagent (cellcounting-Lite2.0, Vazyme, DD1101-02) was added to each well, and the cells were shaken at room temperature for 15 minutes to fully lyse the cells, and then allowed to stand at room temperature for 15 minutes, and the fluorescence intensity was detected. The calculation formula is: % Inhibition = 100-(the signal of the well with the compound to be tested-the signal of the well without cells and containing only the culture medium)/(the signal of the well with cells but without the compound-the signal of the well without cells and containing only the culture medium) × 100.

The dose-effect curve was fitted using the analysis software GraphPad Prism 5 to obtain the IC ₅₀ value of each compound on cell activity.

Table 4 Inhibitory activity of HCT116 WT cells

The test results show that the compounds of the present application have weak inhibitory activity against MTAP wild-type HCT116 cells, but show excellent selectivity against MTAP-deficient OCI-LY19 cells. For example, the IC ₅₀ of Example A16 against OCI-Ly19 cells is 24 nM, and the IC ₅₀ of Example A16 against MTAP wild-type HCT116 cells is 4121 nM, with a selectivity of more than 150 times.

Experimental Example 5: HCT116 MTAP WT and HCT116 MTAP-/- cell proliferation inhibition experiment

The inhibitory effect of the compounds of the present invention on tumor cell proliferation was tested using two cell lines, HCT116 MTAP WT (Nanjing Kebai Biotechnology Co., Ltd., CBP60028) and HCT116 MTAP-/-.

Cell plating:

1) HCT116 MTAP WT and HCT116 MTAP-/- cells were grown to 90% confluence in culture dishes in McCoy's 5A medium (BOSTER, Catalog No. PYG0025) containing 10% FBS (Ecocell, Catalog No. FND500), digested into single cells using trypsin, and the cells were counted.

2) Dilute the cells to the required density with fresh medium. The plating density of HCT116 MTAP WT cells is 120 cells/well, and the plating density of HCT116 MTAP-/- cells is 160 cells/well.

3) Use an electric dispenser to draw 100 μL of cell suspension into a 96-well plate and add 200 μL 1× PBS to the edge wells to prevent evaporation.

Formulated compound:

1) First, the stock solution of the compound was diluted from 10 mM to 4000, 1200, 400, 120, 40, 12, 4, and 1.2 μM, with a DMSO content of 100%.

2) Use an electric gun to take 1.3 μL of the above graded diluted compounds and DMSO (as a control well) and add them to 258.7 μL of culture medium. At this time, the dilution is 200 times and the DMSO content is 0.5%.

3) Use an electric dispenser to take 100 μL of the diluted compound in step 2) and add it to the wells of the plated cells to make the final concentrations of the compound: 10, 3, 1, 0.3, 0.1, 0.03, 0.01, 0.003, 0.001, 0 μM, and set up 2 replicate wells for each concentration. At this time, there are 200 μL of culture medium in each well of the cell culture plate, and the DMSO content is 0.25%.

4) The cell culture plate was returned to the 37°C, 5% CO ₂ incubator and tested 10 days after compound treatment.

CTG test:

1) After 10 days of culture, remove the cells, aspirate all the culture medium, and add 100 μL/well of the pre-prepared detection solution (CTG reagent (cell counting-Lite 2.0, Vazyme, DD1101-03) and culture medium mixed in a ratio of 1:1) using a dispenser.

2) Incubate on a shaker at room temperature for 15 minutes and allow to equilibrate at room temperature for 15 minutes.

3) Use a multifunctional microplate reader (BGM, FSX) was used to detect the fluorescence signal intensity.

Data Analysis:

1) Calculate the cell viability, Cell viability % = As/Ac × 100%. As: fluorescence intensity detection value of the test well (culture medium containing cells, CTG, test compound), Ac: fluorescence intensity detection value of the control well (culture medium containing cells, CTG, DMSO).

2) Using the log value of the compound concentration as the X-axis and the cell viability (Cell viability %) as the Y-axis, a dose-effect curve was fitted to obtain the IC ₅₀ value of each compound's cell proliferation inhibitory activity.

Table 5 Proliferation inhibitory activity of the compounds of the present invention on HCT116 MTAP WT and HCT116 MTAP-/- cells

The test results show that the compounds of the present application have weak inhibitory activity on MTAP wild-type HCT116 cells, but significantly stronger inhibition on MTAP-deficient HCT116 cells, reflecting good selectivity.

Claims (39)

The compound represented by formula (I), its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts:
in, represents any single or double bond allowed by the valence; X ₁ and X ₂ are independently selected from NR ^X , S, N, CR ^X1 ; R ^X and R ^X1 are independently selected from H, halogen, -C ₁ ～C ₆ alkyl, deuterium-substituted -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl, -C(O)-C ₁ ～C ₃ alkyl; X ₃ is selected from N, CR ³ ; R ³ is selected from H, halogen, -C ₁ ～C ₆ alkyl, -C(O)NH ₂ , -C(O)NH(C ₁ ～C ₃ alkyl), -C(O)N(C ₁ ～C ₃ alkyl) ₂ ; X ₄ is selected from N, CR ⁵ ; R ⁷ is selected from H, halogen, -CN, -NO ₂ , -NH ₂ , -C ₁ ～C ₆ alkyl, deuterium-substituted -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl; L is selected from a bond, n is independently selected from 0, 1, and 2; R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ～C ₆ alkyl, -C ₂ ～C ₆ alkenyl, halogen-substituted -C ₁ ～C ₆ alkyl, -OC ₁ ～C ₆ alkyl, 3-6-membered carbocyclic ring, 5-10-membered heteroaromatic ring, 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring, aromatic ring may be optionally substituted by one or more halogen, -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -OC ₁ ～C ₃ alkyl; R ⁴ and R ⁶ are independently selected from hydrogen, -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl, -C ₀ ～C ₂ alkylene-(3-10 membered carbocyclic ring), -C ₀ ～C ₂ alkylene-(4-11 membered heterocyclic ring), -C ₀ ～C ₂ alkylene-(6-10 membered aromatic ring) or -C ₀ ～C ₂ alkylene-(5-10 membered heteroaromatic ring); wherein the alkyl, alkylene, carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ⁴¹ ; R ⁴¹ is selected from hydrogen, halogen, amino, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₆ alkyl, -C ₂ ～C ₆ alkenyl, -C ₂ ～C ₆ alkynyl, halogen-substituted -C ₁ ～C ₆ alkyl, halogen-substituted -C ₂ ～C ₆ alkenyl, halogen-substituted -C ₂ ～C ₆ alkynyl, -OC ₁ ～C ₆ alkyl; R ² is selected from a 3-10-membered carbocyclic ring, a 4-11-membered heterocyclic ring, a 6-10-membered aromatic ring or a 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ²¹ ; ^R21 is selected from hydrogen, halogen, nitro, cyano, oxo, _-NH2 , -C(O) _NH2 , -C(O)H, -C(O)OH, _-C1 ~ _C6 alkyl, _-C2 ~ _C6 alkenyl, _-C2 ~ _C6 alkynyl, halogen-substituted _-C1 ~ _C6 alkyl, halogen-substituted _-C2 ~ _C6 alkenyl, halogen-substituted _-C2 ~ _C6 alkynyl, _-OC1 ~ _C6 alkyl, -OC2~C6 _alkenyl , _-OC2 ~C6 alkynyl, -O _- halogen-substituted _C1 ~ _C6 alkyl, -O-halogen-substituted _C2 ~ _{C6 alkenyl} , -O-halogen-substituted _C2 ~C6 _6- alkynyl, -O-(3-6-membered carbocyclic ring), 3-10-membered carbocyclic ring, 4-11-membered heterocyclic ring, 6-10-membered aromatic ring or 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ^c ; R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ~ C ₆ alkyl, -C ₂ ~ C ₆ alkenyl, -C ₂ ~ C ₆ alkynyl, halogen-substituted -C ₁ ~ C ₆ alkyl, halogen-substituted -C ₂ ~ C ₆ alkenyl, halogen-substituted -C ₂ ~ C ₆ alkynyl, -OC ₁ ~ C ₆ alkyl, -OC ₂ ~ C ₆ alkenyl, -OC ₂ ~ C ₆ alkynyl, -O-halogen-substituted C ₁ ~ C ₆ alkyl, -O-halogen-substituted C ₂ ~ C ₆ alkenyl or -O-halogen-substituted C ₂ ~ C ₆ alkynyl. The compound according to claim 1, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that the compound of formula (I) is as shown in formula (IIa) or formula (IIb):
in, R ¹ , R ² , R ⁷ , ^RX , ^RX1 , L, X ₃ and X ₄ are as described in claim 1. The compound according to claim 1, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that the compound of formula (I) is as shown in formula (IIc), formula (IId), formula (IIe) or formula (IIf):
in, R ¹ , R ² , R ³ , R ⁷ , ^RX , ^RX1 , L and X ₄ are as described in claim 1. The compound according to claim 1, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that the compound of formula (I) is as shown in formula (Ia):
in, represents any single or double bond allowed by the valence; R ¹ , R ² , R ⁵ , X ₁ , X ₂ and L are as described in claim 1. The compound according to any one of claims 1 to 3, its stereoisomer, deuterated substance, solvate, and pharmaceutically acceptable salt, characterized in that: R ⁷ is selected from H, halogen, -CN, -NO ₂ , -NH ₂ , -C ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl substituted with deuterium, -C ₁ ～C ₃ alkyl substituted with halogen. The compound according to any one of claims 1 to 3, its stereoisomer, deuterated substance, solvate, and pharmaceutically acceptable salt, characterized in that: R ³ is selected from H, halogen, -C ₁ -C ₃ alkyl, -C(O)NH ₂ , -C(O)NH(C ₁ -C ₃ alkyl), -C(O)N(C ₁ -C ₃ alkyl) ₂ . The compound according to any one of claims 1 to 3, its stereoisomer, deuterated substance, solvate, and pharmaceutically acceptable salt, characterized in that: X ₄ is selected from N, CR ⁵ ; ^R5 is selected from hydrogen, halogen, _-C1 ~ _C3 alkyl, _-C2 ~ _C4 alkenyl, halogen-substituted _-C1 ~ _C3 alkyl, _-OC1 ~ _C3 alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring and aromatic ring may be optionally substituted by one or more halogen, _-C1 ~ _C3 alkyl, halogen-substituted _-C1 ~ _C3 alkyl and -OC1~ _C3 alkyl _. The compound according to any one of claims 1 to 7, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ^X and R ^X1 are each independently selected from H, halogen, -C ₁ ～C ₃ alkyl, deuterium-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, and -C(O)CH ₃ . The compound according to any one of claims 1 to 8, its stereoisomer, deuterated substance, solvate, pharmaceutically acceptable salt, characterized in that: R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ~ C ₃ alkyl, -C ₂ ~ C ₄ alkenyl, halogen-substituted -C ₁ ~ C ₃ alkyl, -OC ₁ ~ C ₃ alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring and aromatic ring may be optionally substituted by one or more halogen, -C ₁ ~ C ₃ alkyl, halogen-substituted -C ₁ ~ C ₃ alkyl and -OC ₁ ~ C ₃ alkyl. The compound according to claim 1, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that the compound of formula (I) is as shown in formula (Ia):
in, X ₁ and X ₂ are independently selected from NH, S, N, and CH; represents any single or double bond allowed by the valence; L is selected from a bond, n is independently selected from 0, 1, and 2; R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl, -C ₂ ～C ₆ alkenyl, -OC ₁ ～C ₆ alkyl, 3-6-membered carbocyclic ring, 5-10-membered heteroaromatic ring, 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring, aromatic ring may be optionally substituted by one or more halogen, -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -OC ₁ ～C ₃ alkyl; for example, R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl; R ⁴ and R ⁶ are independently selected from hydrogen, -C ₁ ～C ₆ alkyl, halogen-substituted -C ₁ ～C ₆ alkyl, -C ₀ ～C ₂ alkylene-(3-10 membered carbocyclic ring), -C ₀ ～C ₂ alkylene-(4-11 membered heterocyclic ring), -C ₀ ～C ₂ alkylene-(6-10 membered aromatic ring) or -C ₀ ～C ₂ alkylene-(5-10 membered heteroaromatic ring); wherein the alkyl, alkylene, carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ⁴¹ ; R ⁴¹ is selected from hydrogen, halogen, amino, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₆ alkyl, -C ₂ ～C ₆ alkenyl, -C ₂ ～C ₆ alkynyl, halogen-substituted -C ₁ ～C ₆ alkyl, halogen-substituted -C ₂ ～C ₆ alkenyl, halogen-substituted -C ₂ ～C ₆ alkynyl, -OC ₁ ～C ₆ alkyl; R ² is selected from a 3-10-membered carbocyclic ring, a 4-11-membered heterocyclic ring, a 6-10-membered aromatic ring or a 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ²¹ ; ^R21 is selected from hydrogen, halogen, nitro, cyano, oxo, _-NH2 , -C(O) _NH2 , -C(O)H, -C(O)OH, _-C1 ~ _C6 alkyl, _-C2 ~ _C6 alkenyl, _-C2 ~ _C6 alkynyl, halogen-substituted _-C1 ~ _C6 alkyl, halogen-substituted _-C2 ~ _C6 alkenyl, halogen-substituted _-C2 ~ _C6 alkynyl, _-OC1 ~ _C6 alkyl, -OC2~C6 _alkenyl , _-OC2 ~C6 alkynyl, -O _- halogen-substituted _C1 ~ _C6 alkyl, -O-halogen-substituted _C2 ~ _{C6 alkenyl} , -O-halogen-substituted _C2 ~C6 _6- alkynyl, -O-(3-6-membered carbocyclic ring), 3-10-membered carbocyclic ring, 4-11-membered heterocyclic ring, 6-10-membered aromatic ring or 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ^c ; R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ~ C ₆ alkyl, -C ₂ ~ C ₆ alkenyl, -C ₂ ~ C ₆ alkynyl, halogen-substituted -C ₁ ~ C ₆ alkyl, halogen-substituted -C ₂ ~ C ₆ alkenyl, halogen-substituted -C ₂ ~ C ₆ alkynyl, -OC ₁ ~ C ₆ alkyl, -OC ₂ ~ C ₆ alkenyl, -OC ₂ ~ C ₆ alkynyl, -O-halogen-substituted C ₁ ~ C ₆ alkyl, -O-halogen-substituted C ₂ ~ C ₆ alkenyl or -O-halogen-substituted C ₂ ~ C ₆ alkynyl. The compound according to claim 10, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that the compound of formula (Ia) is as shown in formula (Ie):
Wherein, R ¹ , R ² , R ⁵ and L are as described in claim 10. The compound according to claim 10, its stereoisomer, deuterated form, solvate, and pharmaceutically acceptable salt, characterized in that the compound (Ia) is represented by formula (Ib), formula (Ic) or formula (Id):
Wherein, R ¹ , L and R ² are as described in claim 10. The compound according to any one of claims 1 to 12, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ² is selected from a 3-7-membered carbocyclic ring, a 4-6-membered monocyclic heterocyclic ring, a 9-11-membered fused heterocyclic ring, a 6-10-membered aromatic ring, or a 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ²¹ ; ^R21 is selected from hydrogen, halogen, nitro, cyano, oxo, _-NH2 , -C(O) _NH2 , -C(O)H, -C(O)OH, _-C1 ~ _C3 alkyl, _-C2 ~ _C3 alkenyl, _-C2 ~ _C3 alkynyl, halogen-substituted _-C1 ~ _C3 alkyl, halogen-substituted _-C2 ~ _C3 alkenyl, halogen-substituted _-C2 ~ _C3 alkynyl, _-OC1 ~ _C3 alkyl, -OC2~C3 _alkenyl , _-OC2 ~C3 alkynyl, -O _- halogen-substituted _C1 ~ _C3 alkyl, -O-halogen-substituted _C2 ~ _{C3 alkenyl} , -O-halogen-substituted _C2 ~C3 _3- alkynyl, -O-(3-6-membered carbocyclic ring), 3-10-membered carbocyclic ring, 4-11-membered heterocyclic ring, 6-10-membered aromatic ring or 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ^c ; R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, -C ₂ ～C ₄ alkenyl, -C ₂ ～C ₄ alkynyl, halogen-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₂ ～C ₄ alkenyl, halogen-substituted -C ₂ ～C ₄ alkynyl, and -OC ₁ ～C ₃ alkyl. The compound according to claim 13, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ² is selected from wherein R ²¹ is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -OC ₁ ～C ₃ alkyl, -O-halogen-substituted C ₁ ～C ₃ alkyl, -O-(3-6-membered carbocyclic ring), 3-10-membered carbocyclic ring, 4-11-membered heterocyclic ring, 6-10-membered aromatic ring or 5-10-membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ^c ; y = 0, 1, 2 or 3; R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, -C ₂ ～C ₄ alkenyl, -C ₂ ～C ₄ alkynyl, halogen-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₂ ～C ₄ alkenyl, halogen-substituted -C ₂ ～C ₄ alkynyl, and -OC ₁ ～C ₃ alkyl. The compound according to claim 14, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ²¹ is selected from hydrogen, halogen, -OC ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl, -O-halogen substituted C ₁ ～C ₃ alkyl, For example, selected from hydrogen, halogen, -OC ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl, The compound according to any one of claims 1 to 12, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: L is selected from a bond, R ⁴ and R ⁶ are independently selected from hydrogen, -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -C ₀ ～C ₁ alkylene-(3-6 membered carbocyclic ring), -C ₀ ～C ₁ alkylene-(4-6 membered heterocyclic ring), -C ₀ ～C ₁ alkylene-(6-10 membered aromatic ring), -C ₀ ～C ₁ -(5-10 membered heteroaromatic ring); wherein the alkyl, alkylene, carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ⁴¹ ; R ⁴¹ is selected from hydrogen, halogen, amino, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ -C ₃ alkyl, -C ₂ -C ₃ alkenyl, -C ₂ -C ₃ alkynyl, halogen-substituted -C ₁ -C ₃ alkyl, halogen-substituted -C ₂ -C ₆ alkenyl, halogen-substituted -C ₂ -C ₆ alkynyl, and -OC ₁ -C ₆ alkyl. The compound according to claim 16, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ⁴ and R ⁶ are independently selected from hydrogen, -C ₁ to C ₃ alkyl, -C ₁ to C ₃ alkyl substituted with halogen, 3-6-membered carbocyclic ring, -CH ₂ -(3-6-membered carbocyclic ring), 4-6-membered heterocyclic ring, -CH ₂ -(4-6-membered heterocyclic ring), 6-10-membered aromatic ring, -CH ₂ -(6-10-membered aromatic ring), 5-10-membered heteroaromatic ring, -CH ₂ -(5-10-membered heteroaromatic ring); wherein the alkyl, carbocyclic ring, heterocyclic ring, aromatic ring, heteroaromatic ring may be optionally substituted by one or more R ⁴¹ ; R ⁴¹ is selected from hydrogen, halogen, methyl, ethyl, isopropyl. The compound according to claim 17, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ⁴ and R ⁶ are independently selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, The compound according to any one of claims 1 to 11, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ² is selected from R ²¹ is selected from hydrogen, halogen, -OC ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl, -O-halogen substituted C ₁ ～C ₃ alkyl, y = 0, 1, 2 or 3; R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, -C ₂ ～C ₄ alkenyl, -C ₂ ～C ₄ alkynyl, halogen-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₂ ～C ₄ alkenyl, halogen-substituted -C ₂ ～C ₄ alkynyl, -OC ₁ ～C ₃ alkyl; L is selected from a bond, R ⁴ and R ⁶ are independently selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, R ⁴¹ is selected from hydrogen, halogen, methyl, ethyl, isopropyl; R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ~ C ₃ alkyl, -C ₂ ~ C ₄ alkenyl, halogen-substituted -C ₁ ~ C ₃ alkyl, -OC ₁ ~ C ₃ alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring and aromatic ring may be optionally substituted by one or more halogen, -C ₁ ~ C ₃ alkyl, halogen-substituted -C ₁ ~ C ₃ alkyl and -OC ₁ ~ C ₃ alkyl. The compound according to any one of claims 1 to 3, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ² is selected from R ²¹ is selected from hydrogen, halogen, -OC ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl, -O-halogen substituted C ₁ ～C ₃ alkyl, y = 0, 1, 2 or 3; R ^c is selected from hydrogen, halogen, nitro, cyano, oxo, -NH ₂ , -C(O)NH ₂ , -C(O)H, -C(O)OH, -C ₁ ～C ₃ alkyl, -C ₂ ～C ₄ alkenyl, -C ₂ ～C ₄ alkynyl, halogen-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₂ ～C ₄ alkenyl, halogen-substituted -C ₂ ～C ₄ alkynyl, -OC ₁ ～C ₃ alkyl; L is selected from a bond, R ⁴ and R ⁶ are independently selected from hydrogen, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, R ⁴¹ is selected from hydrogen, halogen, methyl, ethyl, isopropyl; R ^X , R ^X1 are independently selected from H, halogen, -C ₁ ～C ₃ alkyl, deuterium-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -C(O)CH ₃ ; X ₄ is selected from N, CR ⁵ ; R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ～C ₃ alkyl, -C ₂ ～C ₄ alkenyl, halogen-substituted -C ₁ ～C ₃ alkyl, -OC ₁ ～C ₃ alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring and aromatic ring may be optionally substituted by one or more halogen, -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl, -OC ₁ ～C ₃ alkyl; R ⁷ is selected from H, halogen, -CN, -NO ₂ , -NH ₂ , -C ₁ ～C ₃ alkyl, deuterium-substituted -C ₁ ～C ₃ alkyl, halogen-substituted -C ₁ ～C ₃ alkyl; R ³ is selected from H, halogen, -C ₁ -C ₃ alkyl, -C(O)NH ₂ , -C(O)NH(C ₁ -C ₃ alkyl), -C(O)N(C ₁ -C ₃ alkyl) ₂ . The compound according to any one of claims 1 to 20, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: L is selected from a bond, The compound according to any one of claims 1 to 11, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: ^R1 and ^R5 are independently selected from hydrogen, halogen, _-C1 - _C3 alkyl, halogen-substituted _-C1 - _C3 alkyl, _-C2 - _C4 alkenyl, _-OC1 - _C3 alkyl, 3-4-membered carbocyclic ring, 5-6-membered heteroaromatic ring or 6-10-membered aromatic ring; wherein the carbocyclic ring, heterocyclic ring, heteroaromatic ring or aromatic ring may be optionally substituted by one or more substituted by halogen, -C ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl substituted by halogen, or -OC ₁ ～C ₃ alkyl; for example, R ¹ and R ⁵ are independently selected from hydrogen, halogen, -C ₁ ～C ₃ alkyl, or -C ₁ ～C ₃ alkyl substituted by halogen. The compound according to any one of claims 1 to 11, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ¹ and R ⁵ are independently selected from hydrogen, methyl, ethyl, Cl, Br, I, F, -CF ₃ , -CH ₂ F, -CHF ₂ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ , -CH ₂ CF ₃ , -Cl, -Br, -OCH ₃ , Phenyl, For example, R ¹ and R ⁵ are each independently selected from hydrogen, methyl, ethyl, F, -CF ₃ , -CH ₂ F, -CHF ₂ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ or -CH ₂ CF ₃ . The compound according to claims 1-20, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ¹ is selected from hydrogen, halogen, -C ₁ ～C ₃ alkyl or halogen-substituted -C ₁ ～C ₃ alkyl. The compound according to claim 24, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: ^R1 is selected from hydrogen, methyl _, ethyl, Cl, Br, I, F, _-CF3 , _-CH2F , _-CHF2 , _{-CH2CH2F , -CH2CHF2 or -CH2CF3 .} The compound according to any one of claims 1 to 20, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ²¹ is selected from H, F, Cl, Br, -CF ₃ , -OCH ₃ , -OCF ₃ , methyl, ethyl, isopropyl, The compound according to any one of claims 1 to 20, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ² is selected from For example, selected from The compound according to claim 12, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ¹ is selected from hydrogen, halogen, -C ₁ ～C ₃ alkyl or halogen-substituted -C ₁ ～C ₃ alkyl; L is selected from R ⁴ is selected from hydrogen, -C ₁ ～C ₃ alkyl, -C ₁ ～C ₃ alkyl substituted with halogen, 3-6 membered carbocyclic ring or 4-6 membered heterocyclic ring; R ² is selected from a 3-7 membered carbocyclic ring, a 4-10 membered heterocyclic ring, a 6-10 membered aromatic ring or a 5-10 membered heteroaromatic ring; wherein the carbocyclic ring, heterocyclic ring, aromatic ring or heteroaromatic ring may be optionally substituted by one or more R ²¹ . The compound according to claim 28, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ¹ is selected from hydrogen, methyl, ethyl, F, Cl, Br, I, -CF ₃ , -CH ₂ F, -CHF ₂ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ or -CH ₂ CF ₃ ; L is selected from ^R4 is selected from methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl or cyclopentyl; R ² is selected from The compound according to claims 1-3, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: ^R7 is selected from H, F, Cl, Br, I, -CN, -NO2, _{-NH2 ,} methyl, ethyl _, propyl, _isopropyl , -CF3, _-CH2F , _{-CHF2 , -CH2CH2F} , _-CH2CHF2 , _-CH2CF3 , _{-CH2D , -CH2CD3 .} The compound according to claims 1-3, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: ^R3 is selected from H, F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, -C(O) _NH2 , -C(O) _NHCH3 , -C(O)N( _CH3 ) ₂ . The compound according to claims 1-4, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that: R ^X and R ^X1 are each independently selected from H, F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, -CF ₃ , -CH ₂ F, -CHF ₂ , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ , -C(O)CH ₃ , -CH ₂ CF ₃ , -CD ₃ , -CH ₂ D, -CH ₂ CD ₃ . The compound according to claim 1, its stereoisomers, deuterated substances, solvates, and pharmaceutically acceptable salts, characterized in that the compound is selected from:





A pharmaceutical composition comprising a therapeutically effective amount of the compound according to any one of claims 1 to 33, its stereoisomers, deuterated substances, solvates, pharmaceutically acceptable salts, and pharmaceutically acceptable carriers and/or excipients. Use of the compound according to any one of claims 1 to 33, its stereoisomer, deuterated form, solvate, pharmaceutically acceptable salt, or the pharmaceutical composition according to claim 34 in the preparation of a medicament for treating a PRMT5-mediated disease. The use according to claim 35, characterized in that the PRMT5-mediated disease is a tumor. The use according to claim 36 is characterized in that the tumor is associated with MTAP gene deletion and/or MTA accumulation. The use according to claim 36 or claim 37, characterized in that the tumor is selected from neuroma, adenocarcinoma, adrenal cancer, anal cancer, angiosarcoma, appendix cancer, benign monoclonal gamma disease, bile duct cancer, bladder cancer, brain cancer, bronchial cancer, carcinoid tumor, cervical cancer, choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer, epithelial cancer, ependymoma, endothelial sarcoma, endometrial cancer, esophageal cancer, Ing's sarcoma, eye cancer, hypereosinophilia, gallbladder cancer, gastric cancer, gastrointestinal stromal tumors, head and neck cancer, oral cancer, leukemia, lymphoma, multiple myeloma, heavy chain disease, hemangioblastoma, inflammatory myofibroblastic tumor, immune cell amyloidosis, kidney cancer, liver cancer, lung cancer, leiomyosarcoma, mastocytosis, myelodysplastic syndrome, mesothelioma, myeloproliferative diseases, essential thrombocythemia, idiopathic extramedullary metaplasia, chronic idiopathic myelofibrosis, hypereosinophilic syndrome, neuroblastoma, neurofibroma, neuroendocrine cancer, osteosarcoma, ovarian cancer, papillary adenocarcinoma, penile cancer, breast cancer, pancreatic cancer, skin cancer, cancer with homozygous deletion of methylthioadenosine phosphorylase gene. The use according to claim 36 or claim 37 is characterized in that the tumor is selected from lymphangiosarcoma, lymphangioendothelioma, hemangioma, meningioma, astrocytoma, oligodendroglioma, medulloblastoma, cervical adenocarcinoma, colon cancer, rectal cancer, colorectal adenocarcinoma, Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcomas, uterine cancer, uterine sarcoma, esophageal adenocarcinoma, Barrett's adenocarcinoma, intraocular melanoma, retinoblastoma, gastric adenocarcinoma, head and neck squamous cell carcinoma, oral squamous cell carcinoma, laryngeal cancer, pharyngeal cancer, nasopharyngeal carcinoma, oropharyngeal cancer, acute lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, chronic neutrophilic leukemia, follicular lymphoma, chronic lymphocytic leukemia, small lymphocytic lymphoma, lymph node marginal zone B cell lymphoma, splenic marginal zone B B-cell lymphoma, primary mediastinal B-cell lymphoma, Burkitt's lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma and primary central nervous system lymphoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma, alpha chain disease, gamma chain disease, mu chain disease, Wilms' tumor, renal cell carcinoma, hepatocellular carcinoma, bronchogenic carcinoma, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, leiomyosarcoma, systemic mastocytosis, neurofibromatosis type 1 or 2, schwannomatosis, gastroenteropancreatic neuroendocrine tumors, cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma, neurofibrosarcoma.