RU2793732C2 - Pde9 inhibitor and its use - Google Patents

Abstract

FIELD: medicine technology.

SUBSTANCE: group of inventions includes a compound of formula (I), or its pharmaceutically acceptable salts or stereoisomers, a pharmaceutical composition and use. In the formula (I), each of X₁, X₂, X₃ and X₄ independently represents CR₃ or N, and X₁ X₂, X₃ and X₄ do not simultaneously represent CR₃ ; R₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, carboxyl, cyano, halogen, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 alkylamino, (C-_1-6 alkyl)₂ amino , C_2-8 alkenyl, C_2-8 alkynyl, C_1-6 alkylsulfonyl, C_1-6 alkylthio, C_3-6 cycloalkyl, 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, piperazinyl and morpholinyl, C_{1 -6} alkylcarbonyl, aminocarbonyl, C_1-6 alkylaminocarbonyl, (C_1-6 alkyl)₂ aminocarbonyl, 4-6 membered heterocyclylcarbonyl selected from piperazinylcarbonyl and azetidinylcarbonyl, and pyrimidinyloxy, where C_1-6 alkyl, C_1-6 alkoxy , (C_1-6 alkyl)₂ amino, C_2-8 alkynyl, 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, piperazinyl and morpholinyl, aminocarbonyl, C_1-6 alkylaminocarbonyl and 4-6 membered heterocyclylcarbonyl, selected of piperazinylcarbonyl and azetidinylcarbonyl are unsubstituted or optionally substituted with one or two groups independently selected from a group consisting of hydroxy, amino, cyano, C_1-6 alkyl, C_1-6 alkoxy, (C_1-6 alkyl)₂ amino, C_1-6 alkylcarbonyloxy, C_3-6 cycloalkyl, halogenated C_1-6 alkyl and a 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl which is unsubstituted or optionally substituted with C_1-6 alkyl; L is a bond, -NH-(CH₂)_t -, and t is 0, 1, 2 or 3; ring A is selected from the group consisting of a 5-7 membered saturated monoheterocyclyl having an N atom as a heteroatom, a 7-11 membered saturated spiroheterocyclyl having 1 or 2 heteroatoms selected from O and N, and an aryl having from 6 to 14 carbon atoms; each R₁ is independently selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C_1-6 alkyl optionally substituted with hydroxy, C_1-6 alkoxy, and a 5-membered heteroaryl containing 1 to 3 heteroatoms selected from S and N; m is 0, 1, 2 or 3; and R₂ is hydrogen; with the proviso that: when X₁ is N and X₂, X₃ and X₄ are each independently CR₃, A is not phenyl; when X₁ and X₃ are each independently N and X₂ and X₄ are each independently CR₃, A is not phenyl; and when X₃ is N and X₁, X₂ and X₄ are each independently CR₃, A is not phenyl.

EFFECT: compound of formula (I) exhibiting the properties of a phosphodiesterase 9 (PDE9) inhibitor, which can be used to obtain a drug for the treatment or prevention of diseases mediated by PDE9.

18 cl, 8 tbl, 101 ex

Description

The present application claims the priority of Chinese Patent Application No. 201710900197.8 entitled "PDE9 INHIBITOR AND USE THEREFOF" filed by China National Intellectual Property Administration on September 28, 2017, Chinese Patent Application No. 201810203538.0 entitled "PDE9 INHIBITOR AND USE THEREFOF" filed by China National Intellectual Property Office on March 13, 2018, and Chinese Patent Application No. 201810871998.0 entitled "PDE9 INHIBITOR AND USE THEREFOF" filed by China National Intellectual Property Office on August 2, 2018, the contents of which are incorporated herein by reference in their entirety.

FIELD OF TECHNOLOGY

The present invention relates to the medical field, and in particular, the invention relates to a phosphodiesterase 9 inhibitor represented by formula (I), or a pharmaceutically acceptable salt, a stereoisomer thereof, and their use.

PRIOR ART

Phosphodiesterases (phosphodiesterase, abbreviated as PDE) belong to a class of proteases that selectively degrade the important second messengers cGMP (abbr. from the English "cyclic guanosine monophosphate", which means "cyclic guanosine monophosphate", abbreviated cGMP) and cAMP (abbr. from English "cyclic adenosine monophosphate", which means "cyclic adenosine monophosphate", abbreviated cAMP), that is, they participate in important physiological processes in the body. Based on gene sequence homology and selectivity for cGMP or cAMP, phosphodiesterases can be classified into 11 species (PDE1-PDE11). An important member of the PDE family is the phosphodiesterase PDE9A, which is highly expressed in the testis, brain, small intestine, skeletal muscle, heart, lung, thymus, and pancreas. Significant studies in recent years, many reports in the available literature and clinical data have shown that PDE9A inhibitors are suitable for the treatment of diseases associated with cognitive disorders caused by disorders of the central nervous system, such as Alzheimer's disease and schizophrenia, and neurodegenerative diseases of the brain.

Both nucleotides, cAMP and cGMP, are important second messengers that play a major role in cell signaling processes. They produce the initial activation of protein kinases: a protein kinase activated by cAMP is called protein kinase A (eng. protein kinase A, abbreviated as PKA), and a protein kinase activated by cGMP is called protein kinase G (eng. protein kinase G, abbreviated PKG). Activated PKA and PKG can phosphorylate various cellular effector proteins such as ion channels, G-protein coupled receptors, structural proteins, and transduction factors. Thus, through this mechanism, cAMP and cGMP can regulate most physiological processes in many organs. At the same time, cAMP and cGMP can also directly affect effector proteins, mediating the processes described above. It is well known that cGMP can directly affect ion acceptors, thus affecting the concentration of ions in cells. Phosphodiesterases (PDEs) hydrolyze cAMP and cGMP cyclic monophosphates and convert them into non-activated AMP and GMP monophosphates.

The human PDE9 phosphodiesterase was first cloned in 1998 and sequenced at the same time; it is a PDE with the highest selectivity known to date for cGMP. The binding constant (Km) of cGMP by phosphodiesterase PDE9 is 170 nM, the value of the binding constant of cAMP by phosphodiesterase PDE9 reaches 230,000 nM, that is, it has a selectivity 1000 times greater. Compared to PDE2A and PDE5A, the catalytic activity of PDE9 cannot be increased by exposure to cGMP because PDE9 does not have a cGMP binding site. Thus, PDE9 inhibitors can increase the base concentration of cGMP.

Conventional PDE inhibitors cannot inhibit human PDE9, and therefore drugs such as IBMX (IBMX, i.e., isobutyl methylxanthine), dipyridamole, SKF94120, rolipram, and vinpocetine do not or only very weakly inhibit PDE9 activity.

There is currently no PDE9 inhibitory drug available in the retail market; only a few inhibitors are in clinical trials and two of them are PDE9 inhibitors, PF-04447943 by Pfizer and BI-409306 by BI. These two compounds are currently in Phase I and II clinical trials.

SUMMARY OF THE INVENTION

One of the objectives of the invention is to provide a class of compounds suitable for use as PDE9 protease inhibitors, or pharmaceutically acceptable salts and stereoisomers thereof, which have high PDE9 protease inhibitory activity and selectivity and drug suitability (e.g., good pharmacokinetics). properties and higher stability in liver microsomes) that can be used to treat or prevent related PDE9-mediated diseases and play an important role in the treatment of diseases associated with cognitive impairment caused by disorders of the central nervous system.

Below are the technical solutions to the problem proposed by the present invention.

The present invention relates to a compound represented by formula (I) or pharmaceutically acceptable salts or stereoisomers thereof:

where each of X ₁ , X ₂ , X ₃ and X ₄ independently represents CR ₃ or N, and X ₁ , X ₂ , X ₃ and X ₄ do not simultaneously represent CR ₃ ;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl ) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4- 6-membered heterocyclyl, C _1-6 alkylcarbonyl, aminocarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, 4-6-membered heterocyclylcarbonyl and 5-6-membered heteroaryloxy, where C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _{1- 6} alkylthio, C _3-6 cycloalkyl, 4-6 membered heterocyclyl, C _1-6 alkylcarbonyl, aminocarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, 4-6 membered heterocyclylcarbonyl and 5-6 -membered heteroaryloxy are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkoxy _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonylamino, C _1-6 alkylsulfonylamino, C _1-6 alkylcarbonyloxy, C _3-6 cycloalkyl, C _2-8 alkynyl , halogenated C _1-6 alkyl, C _2-8 alkenyl, halogenated C _1-6 alkoxy, 4-6 membered heterocyclyl which is unsubstituted or optionally substituted by a substituent, and heteroaryl which is unsubstituted or optionally substituted by a substituent;

the substituent of the above 4-6 membered heterocyclyl optionally has a substituent, and the heteroaryl optionally has a substituent selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl and C _1-6 alkoxy;

L is a bond, -NH-(CH ₂ ) _t -, and t is 0, 1, 2 or 3;

ring A is selected from the group consisting of 3-12 membered heterocyclyl, aryl, 5-10 membered heteroaryl, 3-12 membered cycloalkyl and 3-12 membered cycloalkenyl, wherein the 3-12 membered heterocyclyl comprises a heteroatom selected from one of the following elements: O, S, N, or any combination thereof, and the S atom may optionally be oxidized to S(O) or S(O) ₂ and the 5-10 membered heteroaryl includes a heteroatom selected from one of the following elements : O, S, N or any combination thereof;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, 3-12 membered cycloalkyl, 3-12 -membered cycloalkenyl, 3-12-membered heterocyclyl, aryl and 5-10-membered heteroaryl, where C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, 3-12 membered cycloalkyl, 3-12 membered cycloalkenyl , 3-12 membered heterocyclyl, aryl and 5-10 membered heteroaryl are unsubstituted or optionally substituted with a group selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _{1- 6} alkoxy, C _1-6 alkoxy, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonylamino and C _1-6 alkylsulfonylamino;

m is 0, 1, 2 or 3; And

R ₂ is selected from the group consisting of hydrogen, C _1-6 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, and halogenated C _1-6 alkyl.

In one preferred embodiment, X ₂ is N and each of X ₁ , X ₃ and X ₄ is independently CR ₃ .

In another preferred embodiment, X ₄ is N and X ₁ , X ₂ and X ₃ are each independently CR ₃ .

Some embodiments of the present invention relate to the compound represented by formula (I), or its pharmaceutically acceptable salts or stereoisomers,

Where

each of X ₁ , X ₂ , X ₃ and X ₄ independently represents CR ₃ or N, and X ₁ , X ₂ , X ₃ and X ₄ do not simultaneously represent CR ₃ ;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl ) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4- 6-membered heterocyclyl, C _1-6 alkylcarbonyl, aminocarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, 4-6-membered heterocyclylcarbonyl and 5-6-membered heteroaryloxy, where C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _{1- 6} alkylthio, C _3-6 cycloalkyl, 4-6 membered heterocyclyl, C _1-6 alkylcarbonyl, aminocarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, 4-6 membered heterocyclylcarbonyl and 5-6 -membered heteroaryloxy are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkoxy _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonylamino, C _1-6 alkylsulfonylamino, C _1-6 alkylcarbonyloxy, C _3-6 cycloalkyl, C _2-8 alkynyl , halogenated C _1-6 alkyl, C _2-8 alkenyl and halogenated C _1-6 alkoxy;

the substituent of the above 4-6 membered heterocyclyl optionally has a substituent, and the heteroaryl optionally has a substituent selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl and C _1-6 alkoxy;

L is a bond, -NH-(CH ₂ ) _t -, and t is 0, 1, 2 or 3;

ring A is selected from the group consisting of 3-12 membered heterocyclyl, aryl and 5-10 membered heteroaryl, where the 3-12 membered heterocyclyl comprises a heteroatom selected from one of the following elements: O, S, N, or any combination thereof , and the S atom may optionally be oxidized to S(O) or S(O) ₂ , and the 5-10 membered heteroaryl includes a heteroatom selected from one of the following elements: O, S, N, or any combination thereof;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, amino, cyano, halogen, C _1-6 alkyl, C _1-6 alkoxy, 3-12 membered cycloalkyl, 3-12 membered heterocyclyl, aryl, 5- 10-membered heteroaryl, where C _1-6 alkyl, C _1-6 alkoxy, 3-12-membered cycloalkyl, 3-12-membered heterocyclyl, aryl and 5-10-membered heteroaryl are unsubstituted or optionally substituted with a group selected from the group , consisting of C _1-6 alkyl, C _1-6 alkoxy, 3-12 membered cycloalkyl, 3-12 membered heterocyclyl, aryl and 5-10 membered heteroaryl;

m is 0, 1, 2 or 3; And

R ₂ is hydrogen or C _1-6 alkyl.

In one preferred embodiment, X ₂ is N and each of X ₁ , X ₃ and X ₄ is independently CR ₃ .

In another preferred embodiment, X ₄ is N and X ₁ , X ₂ and X ₃ are each independently CR ₃ .

Some embodiments of the present invention relate to the compound represented by formula (I), or its pharmaceutically acceptable salts or stereoisomers:

Where

each of X ₁ , X ₂ , X ₃ and X ₄ independently represents CR ₃ or N, and X ₁ , X ₂ , X ₃ and X ₄ do not simultaneously represent CR ₃ ;

L is a bond, -NH-(CH ₂ ) _t -, and t is 0, 1, 2 or 3;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl ) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4- 6-membered heterocyclyl, C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl, where C _1-6 alkyl, _1-6 alkoxy, C _1-6 alkylamino, (C _{1 -6} alkyl) ₂ amino, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered heterocyclyl , C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro , halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkoxy C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _3-6 cycloalkyl, C _{1 -6} alkylcarbonylamino, C _1-6 alkylsulfonylamino, C _1-6 alkylcarbonyloxy and unsubstituted or C _1-6 alkyl substituted 4-6 membered heterocyclyl;

ring A is selected from the group consisting of 3-12 membered heterocyclyl, aryl, 5-10 membered heteroaryl, 3-12 membered cycloalkyl and 3-12 membered cycloalkenyl, wherein the 3-12 membered heterocyclyl comprises a heteroatom selected from one of the following elements: O, S, N, or any combination thereof, and the S atom may optionally be oxidized to S(O) or S(O) ₂ and the 5-10 membered heteroaryl includes a heteroatom selected from one of the following elements : O, S, N or any combination thereof;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, 3-12-membered cycloalkyl, 3-12 -membered cycloalkenyl, 3-12-membered heterocyclyl, aryl and 5-10-membered heteroaryl, where C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, 3-12 membered cycloalkyl, 3-12 membered cycloalkenyl , 3-12 membered heterocyclyl, aryl and 5-10 membered heteroaryl are unsubstituted or optionally substituted with a group selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _{1- 6} alkoxy, C _1-6 alkoxy, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonylamino and C _1-6 alkylsulfonylamino;

m is 0, 1, 2 or 3; And

R ₂ is selected from the group consisting of hydrogen, C _1-6 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, and halogenated C _1-6 alkyl.

In one preferred embodiment, X ₂ is N and each of X ₁ , X ₃ and X ₄ is independently CR ₃ .

In another preferred embodiment, X ₄ is N and X ₁ , X ₂ and X ₃ are each independently CR ₃ .

Some embodiments of the present invention relate to the compound represented by formula (II), or its pharmaceutically acceptable salts or stereoisomers:

Where

each of X ₂ , X ₃ and X ₄ independently represents CR ₃ or N, and X ₂ , X ₃ and X ₄ do not simultaneously represent CR ₃ ;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl ) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4- 6-membered heterocyclyl, C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl, where C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered heterocyclyl, C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkoxy C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _3-6 cycloalkyl, C _1-6 alkylcarbonylamino, C _1-6 alkylsulfonylamino, C _1-6 alkylcarbonyloxy and unsubstituted or C _1-6 alkyl substituted 4-6 membered heterocyclyl;

L is a bond, -NH-(CH ₂ ) _t -, and t is 0, 1, 2 or 3;

ring A is selected from the group consisting of 3-12 membered heterocyclyl, aryl, 5-10 membered heteroaryl, 3-12 membered cycloalkyl and 3-12 membered cycloalkenyl, wherein the 3-12 membered heterocyclyl comprises a heteroatom selected from one of the following elements: O, S, N, or any combination thereof, and the S atom may optionally be oxidized to S(O) or S(O) ₂ and the 5-10 membered heteroaryl includes a heteroatom selected from one of the following elements : O, S, N or any combination thereof;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, 3-12-membered cycloalkyl, 3-12 -membered cycloalkenyl, 3-12-membered heterocyclyl, aryl and 5-10-membered heteroaryl, where C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, 3-12 membered cycloalkyl, 3-12 membered cycloalkenyl , 3-12 membered heterocyclyl, aryl and 5-10 membered heteroaryl are unsubstituted or optionally substituted with a group selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _{1- 6} alkoxy, C _1-6 alkoxy, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonylamino and C _1-6 alkylsulfonylamino;

m is 0, 1, 2 or 3; And

R ₂ is selected from the group consisting of hydrogen, C _1-6 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, and halogenated C _1-6 alkyl.

In one preferred embodiment, X ₂ is N and X ₃ and X ₄ are each independently CR ₃ .

In another preferred embodiment, X ₄ is N and X ₂ and X ₃ are each independently CR ₃ .

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

each of X ₂ , X ₃ and X ₄ independently represents CR ₃ or N, and X ₂ , X ₃ and X ₄ do not simultaneously represent CR ₃ ;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, carboxyl, cyano, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered nitrogen-containing heterocyclyl, C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl, where C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _2-8 alkenyl, C _{2- 8} alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6-membered nitrogen-containing heterocyclyl, C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, cyano, halo, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonyloxy, C _3-6 cycloalkyl and 4-6 membered heterocyclyl which is unsubstituted or optionally substituted with C _1-6 alkyl;

L is a bond;

ring A is a 3-12 membered heterocyclyl, where the 3-12 membered heterocyclyl comprises a heteroatom selected from one of the following elements: O, S, N, or any combination thereof, and the S atom may be optionally oxidized to S(O) or S(O) ₂ ;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C _1-6 alkyl, C _1-6 alkoxy and 5-6 membered heteroaryl, where C _1-6 alkyl, C _1-6 alkoxy and 5-6 membered heteroaryl are unsubstituted or substituted with a hydroxy group;

m is 0, 1, or 2; And

R ₂ is hydrogen or C _1-6 alkyl.

In one preferred embodiment, X ₂ is N and X ₃ and X ₄ are each independently CR ₃ .

In another preferred embodiment, X ₄ is N and X ₂ and X ₃ are each independently CR ₃ .

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

X ₂ is N and X ₃ and X ₄ are each independently CR ₃ or N, preferably CR ₃ ;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, cyano, halogen, carboxyl, alkyl, alkoxy, alkylamino, (C _1-4 alkyl) ₂ amino, C _2-6 alkenyl, C _2-6 alkynyl, C _1-4 alkylcarbonyl, alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, C _1-4 alkylsulfonyl, C _1-4 alkylthio, aminocarbonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl, where alkyl, alkoxy, C _1-4 alkylamino , (C _1-4 alkyl) ₂ amino, C _2-6 alkenyl, C _2-6 alkynyl, C _{1-4 alkylcarbonyl, C 1-4 alkylaminocarbonyl} , (C _1-6 alkyl) ₂ aminocarbonyl, C _1-4 alkylsulfonyl , C _1-4 alkylthio, aminocarbonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, cyano, halogen, C _1-4 alkyl, C _{1- 4} alkoxy, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, cyclopropyl, C _1-4 alkylcarbonyloxy and 4-6 membered heterocyclyl which is unsubstituted or optionally substituted with C _1-6 alkyl;

L is a bond;

ring A is a 4-12-membered heterocyclyl, where the 4-12-membered heterocyclyl comprises a heteroatom selected from one or a combination of two of the following atoms: O, S and N, and contains at least one N, and ring A connected to L through an N atom, and the S atom may optionally be oxidized to S(O) ₂ ;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C _1-4 alkyl, C _1-4 alkoxy, pyrazolyl, thiazolyl and triazolyl, where C _1-4 alkyl, C _1-4 alkoxy, pyrazolyl , thiazolyl, and triazolyl are unsubstituted or substituted with a hydroxy group; And

m is 0, 1, or 2.

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

X ₂ is N, X ₃ is CR ₃ and X ₄ is CR ₃ or N, preferably CR ₃ ;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, cyano, halogen, carboxyl, alkyl, alkoxy, alkylamino, (C _1-4 alkyl) ₂ amino, C _2-6 alkenyl, C _2-6 alkynyl, C _1-4 alkylcarbonyl, alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, C _1-4 alkylsulfonyl, C _1-4 alkylthio, aminocarbonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl, where alkyl, alkoxy, C _1-4 alkylamino , (C _1-4 alkyl) ₂ amino, C _2-6 alkenyl, C _2-6 alkynyl, C _{1-4 alkylcarbonyl, C 1-4 alkylaminocarbonyl} , (C _1-6 alkyl) ₂ aminocarbonyl, C _1-4 alkylsulfonyl , C _1-4 alkylthio, aminocarbonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, cyano, halogen, C _1-4 alkyl, C _{1- 4} alkoxy, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, cyclopropyl, C _1-4 alkylcarbonyloxy and 4-6 membered heterocyclyl which is unsubstituted or optionally substituted with C _1-6 alkyl;

L is a bond;

ring A is a 4-12 membered heterocyclyl, wherein the 4-12 membered heterocyclyl comprises a heteroatom selected from one or a combination of two of the following atoms; O, S and N, and contains at least one N, and while the ring A is connected to L through the N atom, and the S atom can be optionally oxidized to S(O) ₂ ;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C _1-4 alkyl, C _1-4 alkoxy, pyrazolyl, thiazolyl and triazolyl, where C _1-4 alkyl, C _1-4 alkoxy, pyrazolyl , thiazolyl, and triazolyl are unsubstituted or substituted with a hydroxy group; And

m is 0, 1, or 2.

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

X ₂ is N, X ₃ is CR ₃ and X ₄ is CR ₃ or N, preferably CR ₃ ;

L is a bond;

ring A is a 4-7-membered monoheterocyclyl, where the 4-7-membered monoheterocyclyl comprises a heteroatom selected from one or a combination of two of the following atoms: O, S and N, and contains at least one N, and while ring A connected to L through an N atom, and the S atom may optionally be oxidized to S(O) ₂ ;

preferably ring A is a saturated 4-7 membered nitrogen-containing monoheterocyclyl, more preferably:

more preferably

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, halogen, C _1-4 alkyl, C _1-4 alkoxy, morpholinyl, C _2-6 alkenyl, C _1-4 alkylcarbonyl, C _1-4 alkylaminocarbonyl, (C _1-4 alkyl) ₂ aminocarbonyl and aminocarbonyl, where C _1-4 alkyl, C _1-4 alkoxy, morpholinyl, C _2-6 alkenyl, C _1-4 alkylcarbonyl, C _1-4 alkylaminocarbonyl, (C _1-4 alkyl) ₂ aminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, C _1-4 alkoxy, cyclopropyl, amino, alkylamino, (C _1-4 alkyl) ₂ amino, and 4-6- a membered heterocyclyl which is unsubstituted or optionally substituted with C _1-4 alkyl;

each R ₁ is independently selected from the group consisting of hydrogen, halogen, C _1-4 alkyl, C _1-4 alkoxy, pyrazolyl, thiazolyl and triazolyl, where C _1-4 alkyl, C _1-4 alkoxy, pyrazolyl, thiazolyl and triazolyl are unsubstituted or substituted with hydroxy; And

m is 0, 1, or 2.

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

X ₂ is N and X ₃ and X ₄ are each independently CR ₃ ;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, halogen, C _1-4 alkyl, C _1-4 alkoxy, C _2-6 alkenyl, C _1-4 alkylcarbonyl, C _1-4 alkylaminocarbonyl and aminocarbonyl, where C _1-4 alkyl, C _1-4 alkoxy, C _2-6 alkenyl, C _1-4 alkylcarbonyl, alkylaminocarbonyl and aminocarbonyl are

unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, C _1-4 alkoxy, cyclopropyl, and 4-6 membered heterocyclyl which is unsubstituted or substituted with C _1-6 alkyl;

L is a bond;

ring A is

each R ₁ is independently selected from the group consisting of hydrogen, C _1-4 alkyl and C _1-4 alkoxy; And

m is 0, 1, or 2.

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

X ₂ is N and X ₃ and X ₄ are each independently CR ₃ ;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, halogen, C _1-4 alkyl and morpholinyl, where C _1-4 alkyl is unsubstituted or substituted with one or more hydroxy groups;

L is a bond;

иring A is

And

each R ₁ is independently selected from the group consisting of pyrazolyl, thiazolyl and triazolyl.

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

L is a bond;

X ₂ is N and X ₃ and X ₄ are independently CR ₃ or N, preferably CR ₃ ;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, cyano, halogen, carboxyl, C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkylcarbonyl, C _2-6 alkynyl, C _1-4 alkylaminocarbonyl, (C _1-4 alkyl) ₂ aminocarbonyl, C _1-4 alkylthio, C _1-4 alkylsulfonyl, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, azetidinyl, morpholinyl, piperazinyl, C _2-6 alkenyl and cyclopropyl, where C _1-4 alkyl, alkoxy, C _1-4 alkylcarbonyl, C _2-6 alkynyl, C _1-4 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, C _1-4 alkylthio, C _{1- 4} alkylsulfonyl, alkylamino, (C _1-4 alkyl) ₂ amino, azetidinyl, morpholinyl, piperazinyl, C _2-6 alkenyl and cyclopropyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, halogen , C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, cyclopropyl and C _1-4 alkylcarbonyloxy;

ring A is a 7-12-membered spiroheterocyclyl, wherein the spiroheterocyclyl comprises one or more heteroatoms selected from the group consisting of O, S, and N and contains at least one N, and ring A is bonded to L through an N atom, and the S atom may optionally be oxidized to S(O) ₂ ; preferably, the 7-12 membered spiroheterocyclyl is a saturated 7-12 membered nitrogen-containing spiroheterocyclyl; more preferably, the saturated 7-12-membered nitrogen-containing spiroheterocyclyl is selected from the group consisting of:

preferably ring A is selected from the group consisting of

и

And

more preferably ring A is selected from the group consisting of

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

each of X ₂ , X ₃ and X ₄ independently represents CR ₃ or N;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, cyano, amino, halogen, carboxyl, C _1-4 alkyl, C _1-4 alkoxy, C _2-6 alkenyl, C _1-4 alkylcarbonyl, C _2-6 alkynyl, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, C _1-4 alkylaminocarbonyl, C _1-4 alkylthio, C _1-4 alkylsulfonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl, where C _1-4 alkyl , C _1-4 alkoxy, C _2-6 alkenyl, C _1-4 alkylcarbonyl, C _2-6 alkynyl, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, C _1-4 alkylaminocarbonyl, C _{1- 4} alkylthio, C _1-4 alkylsulfonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, halogen, C _1-4 alkyl, C _1-4 alkoxy , C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, cyclopropyl and C _1-4 alkylcarbonyloxy;

L is a bond;

иring A is selected from the group consisting of

And

m is 0.

In one preferred embodiment, X ₂ is N and X ₃ and X ₄ are each independently CR ₃ .

In another preferred embodiment, X ₄ is N and X ₂ and X ₃ are each independently CR ₃ .

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

each of X ₂ , X ₃ and X ₄ independently represents CR ₃ or N;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, cyano, halogen, carboxyl, alkyl, alkoxy, alkylamino, (C _1-4 alkyl) ₂ amino, C _2-6 alkenyl, C _2-6 alkynyl, alkylcarbonyl, alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, C _1-4 alkylsulfonyl, C _1-4 alkylthio, aminocarbonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl, where C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, C _2-6 alkenyl, C _2-6 alkynyl, C _1-4 alkylcarbonyl, C _1-4 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, C _1-4 alkylsulfonyl, C _1-4 alkylthio, aminocarbonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, cyano, halogen, C _1-4 alkyl , C _1-4 alkoxy, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, cyclopropyl, C _1-4 alkylcarbonyloxy, 4-6 membered heterocyclyl which is unsubstituted or optionally substituted with C _1-6 alkyl;

L is a bond;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C _1-6 alkyl, C _1-6 alkoxy, pyrazolyl, thiazolyl and triazolyl, where C _1-6 alkyl, C _1-6 alkoxy, pyrazolyl , thiazolyl, and triazolyl are unsubstituted or substituted with a hydroxy group;

m is 0, 1 or 2;

ring A is a group selected from the group consisting of:

preferably ring A is selected from the group consisting of

In one preferred embodiment, X ₂ is N and X ₃ and X ₄ are each independently CR ₃ .

In another preferred embodiment, X ₄ is N and X ₂ and X ₃ are each independently CR ₃ .

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

X ₂ is N and X ₃ and X ₄ are each independently CR ₃ or N;

L is -NH-(CH ₂ ) _t - or a bond, and t is 0, 1 or 2;

ring A is phenyl;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, carboxyl, cyano, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _2-8 alkenyl, C _2-6 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered nitrogen-containing heterocyclyl, C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl, where C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _2-8 alkenyl, C _{2- 6} alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered nitrogen heterocyclyl, C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, cyano, halo, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonyloxy, C _3-6 cycloalkyl and 4-6 membered heterocyclyl which is unsubstituted or substituted with C _1-6 alkyl;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl and C _1-6 alkoxy, where C _1-6 alkyl and C _1-6 alkoxy are unsubstituted or optionally substituted with a group selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl and C _1-6 alkoxy;

R ₂ is hydrogen or C _1-6 alkyl; And

m is 0, 1, or 2.

Some embodiments of the present invention relate to a compound represented by formula (I) or (II), or its pharmaceutically acceptable salts or stereoisomers,

Where

X ₂ is N and X ₃ and X ₄ are each independently CR ₃ or N;

L is a bond;

R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, carboxyl, cyano, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _2-8 alkenyl, C _2-6 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered nitrogen heterocyclyl, C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl, where C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _2-8 alkenyl, C _{2- 6} alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered nitrogen heterocyclyl, C _1-6 alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or more groups independently selected from the group consisting of hydroxy, amino, cyano, halo, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonyloxy, C _3-6 cycloalkyl and 4-6 membered heterocyclyl which is unsubstituted or substituted with C _1-6 alkyl;

L is a bond;

ring A is a 5-10 membered heteroaryl, and the 5-10 membered heteroaryl comprises a heteroatom selected from one of the following: O, S, N, or any combination thereof;

each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl, C _1-6 alkoxy, phenyl, and 5-6 membered heteroaryl, where C _1-6 alkyl, C _1-6 alkoxy, phenyl and 5-6 membered heteroaryl are unsubstituted or optionally substituted with a group selected from the group consisting of hydroxy, amino, carboxyl, cyano, nitro, halogen, C _1-6 alkyl and C _{1- 6} alkoxy;

m is 0, 1 or 2; And

R ₂ is hydrogen or C _1-6 alkyl.

Preferably, ring A is a 9-10 membered heteroaryl.

More preferably, ring A is a 9-10 membered nitrogen-containing heteroaryl.

Most preferably, ring A is

One of the embodiments of the present invention relates to the above compounds represented by formulas (I) or (II), or their pharmaceutically acceptable salts or stereoisomers, which are shown in Table 1:

The present invention also relates to a pharmaceutical composition comprising a compound represented by formula (I) or (II), or pharmaceutically acceptable salts or stereoisomers thereof, and one or more additional therapeutically active agents.

In one of the specific embodiments of the present invention, the composition can be used for the combined administration of a "therapeutically effective amount" of a compound represented by formulas (I) or (II), or its pharmaceutically acceptable salts or stereoisomers, together with one or more additional therapeutically active agents, such as sequential administration, simultaneous administration, or administration of a compound formulation comprising a compound of the invention, or pharmaceutically acceptable salts or stereoisomers thereof, and an additional therapeutically active agent.

The present invention also relates to a pharmaceutical composition comprising a compound represented by formulas (I) or (II), or pharmaceutically acceptable salts or stereoisomers thereof.

In some embodiments of the invention, the pharmaceutical composition may include one or more pharmaceutically acceptable carriers.

A pharmaceutically acceptable carrier according to the present invention may be one or more solid or liquid excipients or gel-like materials suitable for human consumption. The pharmaceutically acceptable carrier is preferably of sufficient purity and sufficiently low toxicity, is compatible with the compound or its pharmaceutically acceptable salts or stereoisomers of the present invention, and does not significantly impair their pharmacological action. For example, a pharmaceutically acceptable carrier may be a filler, a binder, a disintegrant, a lubricant, an aqueous solvent or a non-aqueous solvent, and the like.

The pharmaceutical composition of the present invention may be formulated into any pharmaceutically acceptable unit dosage form and administered to a patient or individual in need of such therapy by any suitable means of administration, such as oral, parenteral, rectal or inhalation administration. For oral administration, the formulation may be in the form of tablets, capsules, pills, granules, and the like. For parenteral administration, the formulation may be prepared as an injection solution, a sterile injection powder, and the like.

The invention also relates to the use of a compound represented by formulas (I) or (II), or pharmaceutically acceptable salts or stereoisomers thereof, a pharmaceutical composition or a pharmaceutical composition for the preparation of a medicament for the treatment or prevention of a disease mediated by PDE9. In particular, a PDE9 mediated disease is a cognitive disorder caused by a disorder of the central nervous system. In particular, cognitive impairment includes: impaired perception, attention, memory, and learning ability, examples of which include, without limitation, Alzheimer's disease, schizophrenia, senile memory loss, vascular dementia, traumatic brain injury, stroke, post-stroke dementia, post-traumatic dementia , general attention impairment, attention impairment in children aggravated by problems related to learning and memory, Alzheimer's disease, Lewy body dementia, fronto-lobar dementia, corticobasal degeneration, amyotrophic lateral sclerosis, Huntington's chorea, multiple sclerosis, degeneration of the thalamus, Creutzfeldt-Jakob dementia, HIV dementia, schizophrenia, Korsakoff's psychosis, or depression or bipolar disorder.

The invention also relates to a compound represented by formulas (I) or (II), or to pharmaceutically acceptable salts or stereoisomers thereof, a pharmaceutical composition or pharmaceutical composition intended for use in the treatment or prevention of a disease.

The invention also relates to the use of a compound represented by formulas (I) or (II), or pharmaceutically acceptable salts or stereoisomers thereof, a pharmaceutical composition or pharmaceutical formulation for use in the treatment or prevention of a PDE9 mediated disease. In particular, a PDE9 mediated disease is a cognitive disorder caused by a disorder of the central nervous system. In particular, cognitive impairment includes: impaired perception, attention, memory, and learning ability, examples of which include, without limitation, Alzheimer's disease, schizophrenia, senile memory loss, vascular dementia, traumatic brain injury, stroke, post-stroke dementia, post-traumatic dementia , general attention impairment, attention impairment in children aggravated by problems related to learning and memory, Alzheimer's disease, Lewy body dementia, fronto-lobar dementia, corticobasal degeneration, amyotrophic lateral sclerosis, Huntington's chorea, multiple sclerosis, degeneration of the thalamus, Creutzfeldt-Jakob dementia, HIV dementia, schizophrenia, Korsakoff's psychosis, or depression or bipolar disorder.

The invention also relates to a method for the treatment or prevention of diseases. The method includes administering to a patient in need of appropriate therapy a therapeutically effective amount of a compound represented by formulas (I) or (II), or pharmacologically acceptable salts or stereoisomers thereof, a pharmaceutical composition or a pharmaceutical composition. The disease is a PDE9 mediated disease. In particular, a PDE9 mediated disease is a cognitive disorder caused by a disorder of the central nervous system. In particular, cognitive impairment includes: impaired perception, attention, memory, and learning ability, examples of which include, without limitation, Alzheimer's disease, schizophrenia, senile memory loss, vascular dementia, traumatic brain injury, stroke, post-stroke dementia, post-traumatic dementia , general attention impairment, attention impairment in children aggravated by problems related to learning and memory, Alzheimer's disease, Lewy body dementia, fronto-lobar dementia, corticobasal degeneration, amyotrophic lateral sclerosis, Huntington's chorea, multiple sclerosis, degeneration of the thalamus, Creutzfeldt-Jakob dementia, HIV dementia, schizophrenia, Korsakoff's psychosis, or depression or bipolar disorder.

INFORMATION CONFIRMING THE POSSIBILITY OF IMPLEMENTING THE INVENTION

As used herein, the term "halogen" means fluorine, chlorine, bromine, iodine and the like, preferably fluorine and chlorine.

As used herein, the term "halogenated" means that any hydrogen atom present in a substituent may be replaced by one or more halogen atoms, which may be the same or different. The term "halogen" is defined above.

The term "C _1-6 alkyl" as used herein means a straight or branched alkyl group obtained by removing one hydrogen atom from a hydrocarbon species containing from 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n- butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 1-methyl-2-methylpropyl, etc. d. The term "C _1-4 alkyl" refers to the above examples containing from 1 to 4 carbon atoms.

As used herein, the term "C _2-8 alkenyl" means a straight or branched or cyclic alkenyl group obtained by removing one hydrogen atom from an olefin containing from 2 to 8 carbon atoms and containing a carbon-carbon double bond such as vinyl, 1- propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1,3-butadienyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 1, 4-hexadienyl, etc.

As used herein, the term "C _2-8 alkynyl" means a straight or branched alkynyl group obtained by removing one hydrogen atom from an alkyne containing from 2 to 8 carbon atoms and containing a carbon-carbon triple bond such as ethynyl, propynyl, 2- butynyl, 2-pentynyl, 3-pentynyl, 4-methyl-2-pentynyl, 2-hexynyl, 3-hexynyl, etc.

As used herein, the term "C _1-6 alkoxy" means a group in which "C _1-6 alkyl", as defined above, is connected to the parent particle through an oxygen atom, i.e., the group "C _1-6 alkyl-O- such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, n-pentyloxy, neopentyloxy, n-hexyloxy, etc. The term "C _1-6 alkoxy" refers to the above examples containing from 1 to 4 carbon atoms, that is, the group "C _1-6 alkyl-O-".

The terms used in this work "C _1-6 alkylamino", "(C _1-6 alkyl) ₂ amino", "C _1-6 alkylcarbonylamino", "C _1-6 alkylsulfonylamino", "C _1-6 alkylaminocarbonyl", " (C _1-6 alkyl) ₂ aminocarbonyl", "C _1-6 alkoxycarbonyl", "C _1-6 alkylsulfonyl", "C _1-6 alkylthio", "C _1-6 alkylcarbonyl" mean, respectively, groups C _{1- 6} alkyl-NH-, (C _1-6 alkyl) (C _1-6 alkyl) N-, C _1-6 alkyl-C (O) -NH-, C _1-6 alkyl-S (O) ₂ -NH ₂ -, C _1-6 alkyl-NH-C (O) -, (C _1-6 alkyl) (C _1-6 alkyl) N-C (O) -, C _1-6 alkyl-O-C (O )-, C _1-6 alkyl-S(O) ₂ -, C _1-6 alkyl-S-, C _1-6 alkyl-C(O)-. The term "C _1-6 alkyl" is defined above and is preferably "C _1-4 alkyl".

As used herein, the term "fused ring structure" means a polycyclic structure formed by two or more ring structures connected as an ortho-fused, spirocyclic or bridged ring system. An ortho-fused ring means a fused ring structure formed by two or more ring structures containing two common adjacent atoms belonging to different rings (ie, containing one common bond). Bridged ring means a fused ring structure formed by two or more ring structures containing two common non-adjacent atoms belonging to different rings. A spirocyclic structure is a condensed cyclic structure formed by two or more cyclic structures containing one atom common to different cycles.

Unless otherwise indicated, the term "3-12-membered cycloalkenyl" as used herein includes all monocyclic and fused rings (including fused rings in the form of ortho-fused rings, spirocyclic compounds and bridged cyclic compounds) that can be formed, such as 3-8 membered monocycloalkenyl, 7-11 membered spirocycloalkenyl, 7-11 membered ortho-fused cycloalkenyl, bridged 6-11 membered cycloalkenyl and the like.

As used herein, the term "cycloalkyl" includes all monocyclic systems and fused ring systems (including fused rings in the form of ortho-fused rings, spirocyclic compounds, and bridged ring compounds) that can be formed. For example, "3-12 membered cycloalkyl" may be a monocyclic, bicyclic, or polycyclic cycloalkyl system (also referred to as a fused ring system). Unless otherwise indicated, a monocyclic system is a cyclic hydrocarbon group containing 3 to 8 carbon atoms. Examples of 3-8 membered cycloalkyls include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. Fused cycloalkyl includes ortho-fused cycloalkyl, bridged cycloalkyl, and spirocycloalkyl. The ortho-fused cycloalkyl may be 6-11 membered ortho-fused cycloalkyl and 7-10 membered ortho-fused cycloalkyl, and representative examples of such cycles include, without limitation, bicyclo[3.1.1]heptane, bicyclo[2 ,2,1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane and bicyclo[4.2.1]nonane. Spirocycloalkyl can be 7-12 membered spirocycloalkyl and 7-11 membered spirocycloalkyl, and examples of such systems include, without limitation:

The bridged cycloalkyl may be 6-11 membered bridged cycloalkyl and 7-10 membered bridged cycloalkyl, and examples of such systems include, without limitation:

The term "heterocyclyl" as used herein means a 3-12 membered non-aromatic cyclic group in which at least one carbon atom in the ring is substituted by a heteroatom selected from O, S and N, preferably substituted by 1-3 heteroatoms, wherein a carbon atom, a nitrogen atom and a sulfur atom can be oxidized.

The term "3-12 membered heterocyclyl" means a monocyclic heterocyclyl, bicyclic heterocyclyl or polycyclic heterocyclyl system (also referred to as a fused ring system) comprising saturated and partially saturated heterocyclyl, excluding aromatic rings. Unless otherwise stated, this definition includes all monocyclic rings, fused rings (including fused rings as ortho-fused rings, spirocyclic bridged ring systems), saturated rings, and partially saturated rings that may be formed.

Monoheterocyclyl may be 3-8 membered heterocyclyl, 3-8 membered saturated heterocyclyl, 3-6 membered heterocyclyl, 4-7 membered heterocyclyl, 5-7 membered heterocyclyl, 5-6 membered heterocyclyl, 5-6 oxygen-containing heterocyclyl, 3-8-membered nitrogen-containing heterocyclyl, 5-6-membered nitrogen-containing heterocyclyl, 5-6-membered saturated heterocyclyl, and the like. Examples of "3-8 membered saturated heterocyclyl" include, without limitation, aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydrothiophenyl, imidazolidinyl, pyrazolidinyl, 1,2-oxazolidinyl, 1,3-oxazolidinyl, 1 ,2-thiazolidinyl, 1,3-thiazolidinyl, tetrahydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, piperidinyl, piperazinyl, morpholinyl, 1,4-dioxanyl, 1,4-oxathianyl. Examples of "3-8 membered partially saturated heterocyclyl" include, without limitation, 4,5-dihydroisooxazolyl, 4,5-dihydrooxazolyl, 2,5-dihydrooxazolyl, 2,3-dihydrooxazolyl, 3,4-dihydro-2H-pyrrolyl, 2,3-dihydro-1H-pyrrolyl, 2,5-dihydro-1H-imidazolyl, 4,5-dihydro-1H-imidazolyl, 4,5-dihydro-1H-pyrazolyl, 4,5-dihydro-3H-pyrazolyl, 4,5-dihydrothiazolyl, 2,5-dihydrothiazolyl, 2H-pyranyl, 4H-pyranyl, 2H-thiopyranyl, 4H-thiopyranyl, 2,3,4,5-tetrahydropyridyl, 1,2-isooxazinyl, 1,4-isooxazinyl, or 6H-1,3-oxazinyl and the like. The fused heterocyclic ring includes ortho-fused heterocyclyl, spiroheterocyclyl, bridged heterocyclyl, and may be saturated, partially saturated, or unsaturated, but not aromatic. The fused heterocyclyl is a 5-6 membered monocyclic heterocyclic ring fused to a benzene ring, a 5-6 membered monocyclic cycloalkyl, a 5-6 membered monocyclic cycloalkenyl, a 5-6 membered monocyclic heterocyclyl, or a 5-6 membered monocyclic heteroaryl. The ortho-fused heterocyclyl may be 6-12 membered ortho-fused heterocyclyl, 7-10 membered ortho-fused heterocyclyl, 6-10 membered ortho-fused heterocyclyl, and 6-12 membered saturated ortho-fused heterocyclyl, representative examples which include, without limitation, 3-azabicyclo[3.1.0]hexyl, 3,6-diazabicyclo[3.2.0]heptyl, 3,8-diazabicyclo[4.2.0]octyl, 3,7- diazabicyclo[4,2,0]octyl, octahydropyrrolo[3,4-c]pyrrolyl, octahydropyrrolo[3,4-b]pyrrolyl, octahydropyrrolo[3,4-b][1,4]oxazinyl, octahydro-1H-pyrrolo [3,4-c]pyridinyl, 2,3-dihydrobenzofuran-2-yl, 2,3-dihydrobenzofuranyl-3-yl, indolin-1-yl, indolin-2-yl, indolin-3-yl, 2,3 -dihydrobenzothiophen-2-yl, octahydro-1H-indolyl, octahydrobenzofuranyl. Spiroheterocyclyl may be 6-12 membered spiroheterocyclyl, 7-11 membered spiroheterocyclyl, 7-11 membered saturated spiroheterocyclyl, 6-12 membered saturated spirocyclyl, and examples of such groups include, without limitation:

The bridged heterocyclyl may be 6-12 membered bridged heterocyclyl, 7-11 membered bridged heterocyclyl, and 6-12 membered saturated bridged heterocyclyl, and examples of such groups include, without limitation:

The term "aryl" as used herein means a cyclic aromatic group of 6 to 14 carbon atoms, including phenyl, naphthalene, phenanthrene, and the like.

As used herein, the term "heteroaryl" includes all monocyclic rings, fused rings, and all aromatic and partially aromatic systems that may be formed. For example, "5-10 membered heteroaryl" means an aromatic cyclic group in which at least one carbon atom in the ring is replaced by a heteroatom selected from O, S and N, preferably the ring contains from 1 to 3 heteroatoms, and when this carbon atom or sulfur atom may be oxidized, for example, the carbon atom is substituted with a C(O) group, and the sulfur atom is substituted with an S(O) or S(O) _{2 group} . Heteroaryl includes monocyclic heteroaryl and fused heteroaryl. Unless otherwise indicated, monocyclic heteroaryl may be 5-7 membered heteroaryl or 5-6 membered heteroaryl, and examples of monocyclic heteroaryl include, without limitation, furanyl, imidazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, oxazolyl, pyridyl, pyridazinyl , pyrimidinyl, pyrazinyl, pyrazolyl, pyrrolyl, tetrazolyl, thiadiazolyl, thienyl, triazolyl and triazinyl. In some embodiments, fused heteroaryl means a group in which a monocyclic heteroaryl is fused to phenyl, cycloalkenyl, heteroaryl, cycloalkyl, or heterocyclyl. The fused heteroaryl may be 8-12 membered ortho-fused heteroaryl or 9-10 membered ortho-fused heteroaryl. Examples of fused heteroaryls include, without limitation, benzimidazolyl, benzofuranyl, benzothienyl, benzoxadiazolyl, benzothiadiazolyl, benzothiazolyl, cinnolinyl, 5,6-dihydroquinolin-2-yl, 5,6-dihydroisoquinolin-1-yl, furopyridinyl, indazolyl, indolyl, isoindolyl, isoquinolyl, naphthyridinyl, purinyl, quinolyl, 5,6,7,8-tetrahydroquinol-2-yl, 5,6,7,8-tetrahydroquinolyl, 5,6,7,8-tetrahydroquinol-4-yl, 5,6, 7,8-tetrahydroisoquinol-1-yl, thienopyridinyl, 4,5,6,7-tetrahydro[c][1,2,5]oxadiazolyl and 6,7-dihydro[c][1,2,5]oxadiazol- 4(5H)keto.

As used herein, the term "pharmaceutically acceptable salts" refers to pharmaceutically acceptable addition salts (addition salts) and solvates of acids and bases. Pharmaceutically acceptable salts include salts of acids such as hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, sulfurous acid, formic acid, toluenesulfonic acid, methanesulfonic acid, nitric acid, benzoic acid, citric acid, tartaric acid, maleic acid, hydroiodic acid. , alkanoic acid (such as acetic acid, HOOC-(CH ₂ ) _n -COOH (where n is 0 to 4)), and the like. Base salts include sodium, potassium, calcium, ammonium and similar base salts. Many different non-toxic, pharmaceutically acceptable addition salts are known to those skilled in the art.

"Stereoisomer" of the compound of the present invention having the formula (I) means the enantiomer when the compound having the formula (I) contains an asymmetric carbon atom, the cis-trans isomer when the compound contains a carbon-carbon double bond or a cyclic structure, a tautomer if a ketone or oxime group is present in the compound. Included within the scope of the invention are enantiomers, diastereomers, racemic isomers, cis-trans isomers, tautomers, geometric isomers, epimers of the compound of formula (I), and mixtures thereof.

As used herein, the term "therapeutically effective amount" means that amount of the above compound, or a pharmaceutically acceptable salt, stereoisomer, composition, or pharmaceutical composition thereof, which can at least alleviate the symptoms of the patient's condition after administration to the patient. The actual amount, including "therapeutically effective amount", may vary depending on a number of circumstances, which include, without limitation, the particular condition being treated, the severity of the condition, the physical condition and health of the patient, and the route of administration. Experienced medical professionals can successfully determine the appropriate amount, using methods known in medicine.

DESCRIPTION OF EXAMPLES OF CARRYING OUT THE INVENTION

Abbreviations and designations used in this work mean the following: "DMF" means dimethylformamide; "CDI" means N,N'-carbonyldiimidazole (from the English "N,N'-carbonyldiimidazole"); "DIPEA" means N,N-diisopropylethylamine (from the English "N,N-diisopropylethylamine"); "EA" means ethyl acetate; "PE" means petroleum ether; "DIBAL-H" means diisobutylaluminum hydride (from the English "diisobutylaluminum hydride"); "THF" means tetrahydrofuran; "DHM" means dichloromethane; "TBAF" means tetrabutylammonium fluoride (from the English "tetrabutylammonium fluoride"); "DMAP" means 4-dimethylaminopyridine; "HATU" stands for 2-(7-oxobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate ); "AD-mix-β" means a mixture containing 0.0016 mol (DHQD) 2PHAL (hydrogenated quinidine 1,4-(2,3-naphthyridine) diester -naphthyridine)diether"), 0.4988 mol of powdered potassium carbonate, 0.4988 mol of potassium ferricyanide and 0.0007 mol of potassium osmate dihydrate; "DMAA" means dimethylacetamide; "MTBE" means methyl tert-butyl ether; "Boc" means tert-butyloxycarbonyl (from the English "tert-butyloxycarbonyl"); "TFA" means trifluoroacetic acid; "Xphos" means 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (from "2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl"); "DAST" means diethylaminosulfur trifluoride (from the English "diethylaminosulfur trifluoride"); "LiHMDS" means bis-trimethylsilylamine lithium (from the English "bistrimethylsilylamine lithium"); "TMSCF3" stands for trifluoromethyltrimethylsilane (from the English "trifluoromethyl trimethylsilane").

Receiving example 1

Synthesis of intermediate 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Step 1: Synthesis of 2H-pyrido[3,4-d][1,3]oxazine-2,4(1H)-dione

The starting material, 3-aminoisonicotinic acid (1000 mg, 7.240 mmol, 1.0 eq.) was dissolved in DMF (20 ml). The solution was cooled to 0°C, after which CDI (2000 mg, 12.334 mmol, 1.7 eq.) was added in portions. The system was slowly warmed to room temperature, the reaction was left to proceed overnight, and the completion of the reaction was determined by the method of LC-MS (liquid chromatography with mass spectrometry). The reaction solution was cooled to room temperature, and the resulting reaction solution was directly used in the next step without treatment.

Step 2: Synthesis of 4-hydroxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

To the reaction solution containing the intermediate 2H-pyrido[3,4-d][1,3]oxazine-2,4(1H)-dione (theoretically calculated amount 1188 mg, 7.240 mmol, 1 eq.) obtained in the above step, ethyl cyanoacetate (819 mg, 7.240 mmol) and triethylamine (1581 mg, 14.480 mmol, 2 eq.) were added. The mixture was heated to 150° C. and the reaction was allowed to proceed for about 4 hours, the completion of the reaction was determined by the LC-MS method. The reaction solution was cooled to 50° C. and concentrated to dryness under reduced pressure to give a red semi-solid. The semi-solid was cooled to 10° C. whereupon water (5 ml) was added and stirred. The pH of the system was adjusted to 1 by adding hydrochloric acid (1 mol/l). The mixture was stirred for 15 minutes and filtered with suction. The filter cake was washed with water, aspirated and dried at 40° C. under reduced pressure to give 4-hydroxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (889 mg, 66% yield).

Step 3: Synthesis of 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4-hydroxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (789 mg, 4.216 mmol, 1 eq.), phosphorus oxychloride (2908 mg, 18.970 mmol, 4, 5 eq.) and phosphorus pentachloride (1756 mg, 8.431 mmol, 2 eq.). The system was heated to 100°C, and the reaction was carried out for 1 hour; the completion of the reaction was determined by the LC-MS method. The reaction solution was cooled to room temperature, carefully added dropwise to ice until a large amount of solid precipitated, and then filtered with suction. The filter cake was washed with water and dried under reduced pressure at 40°C to give 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (515 mg, yield: 60.0%) .

Receiving Example 2

Synthesis of intermediate 4,6-dichloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Step 1: Synthesis of 6-chloro-2H-pyrido[3,4-d][1,3]oxazine-2,4(1H)-dione

5-Amino-2-chloroisonicotinic acid (30 g, 0.1738 mol, 1.0 eq.) was dissolved in N,N-dimethylformamide (300 ml) and then N,N'-carbonyldiimidazole (48 g, 0.2955 mol, 1.7 eq.). The system was slowly warmed to room temperature and the reaction was allowed to proceed overnight; the completion of the reaction was determined by the LC-MS method. The reaction solution was cooled to room temperature and injected directly into the next step without treatment.

Step 2: Synthesis of 6-chloro-4-hydroxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Triethylamine (35.182 g, 0.3478 mol, 2 eq.) and ethyl cyanoacetate (19.665 g, 0.1738 mol) were added to the reaction solution obtained above. The reaction in the system was left to proceed at 150°C for 3 hours; the completion of the reaction was determined by the LC-MS method. The reaction solution was cooled to room temperature and concentrated under reduced pressure. Water (200 ml) was added and the pH of the system was adjusted to 1 by adding hydrochloric acid (1 mol/l). The mixture was stirred for 15 minutes and filtered with suction. The filter cake was washed twice with EA and dried at 40° C. to give a light, brick red solid (25.655 g, yield: 66%).

Step 3: Synthesis of 4,6-dichloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-Chloro-4-hydroxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (5.0 g, 0.0226 mol, 1 eq.) and phosphorus oxychloride ( 15 ml). The reaction flask was placed in an oil bath which was heated to 100° C. and allowed to react for 6 minutes. The solid began to slowly dissolve and the pale yellow color began to gradually darken. The completion of the reaction was determined by LC-MS. The system was cooled to room temperature and the appropriate amount of DCM was added to the flask. The system was then poured into ice water (100 ml) and stirred for 10 minutes. The mixture was filtered with suction, the filter cake washed with methyl tert-butyl ether, suctioned off and dried in vacuo at 40° C. to give a pale yellow solid. A total of 25.655 g (0.1157 mol) of 6-chloro-4-hydroxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile was reacted in five portions to give 19.486 g of product (yield: 70 ,1%).

Example 1

Synthesis of 4-(azepan-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 3)

The starting material, 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (19 mg, 0.092 mmol, 1.0 eq.) was dissolved in DMF (0.7 ml), after to which were added azepan (17.4 mg, 0.204 mmol, 1.4 eq.) and DIPEA (48 mg, 0.370 mmol, 4 eq.). The mixture was heated to 100° C. and the reaction was allowed to proceed for 4 hours; the completion of the reaction was determined by the LC-MS method. The reaction solution was cooled to room temperature, whereby a solid precipitated, which was suction filtered. The filter cake was washed with water (20 ml), aspirated and dried in vacuo to give 4-(azepan-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (10.02 mg, 27.0%).

¹ H NMR (400 MHz, DMSO-d ₆ (DMSO - short for "dimethyl sulfoxide")) δ (ppm): 11.96 (s, 1H), 8.64 (s, 1H), 8, 31-8.32 (d, 1H), 7.75-7.77 (d, 1H), 3.80-3.83 (t, 4H), 1.84 (s, 4H), 1.71 ( s, 4H).

Molecular formula: C ₁₅ H ₁₆ N ₄ O, Molecular weight: 268.32, LC-MS (Pos, m/z)=269.2 [M+H ⁺ ].

Example 2

Synthesis of 2-oxo-4-(piperidin-1-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 4)

4-Chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (30 mg, 0.146 mmol, 1.0 eq.) was dissolved in DMF (1 ml), after which piperidine (17 .4 mg, 0.204 mmol, 1.4 eq.) and DIPEA (75.4 mg, 0.584 mmol, 4 eq.). The system was heated to 100°C and left to react for 1.5 hours; the completion of the reaction was determined by the LC-MS method. The reaction solution was cooled and filtered with suction. The filter cake was washed with water and dried in vacuo to give a yellow solid (10.02 mg, yield: 27.0%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.99 (s, 1H), 8.64 (s, 1H), 8.33-8.35 (d, 1H) , 7.58-7.59 (d, 1H), 3.60 (s, 4H), 1.72-1.76 (m, 6H).

Molecular formula: C ₁₄ H ₁₄ N ₄ O, Molecular weight: 254.29, LC-MS (Pos, m/z)=255.1 [M+H ⁺ ].

Example 3

Synthesis of 4-(benzylamino)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 10)

The starting material, 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (30 mg, 0.146 mmol, 1.0 eq.) was dissolved in DMF (0.7 ml), after to which were added benzylamine (22 mg, 0.204 mmol, 1.4 eq.) and DIPEA (75.4 mg, 0.584 mmol, 4 eq.). The system was heated to 100°C and left to react for 1.5 hours; the completion of the reaction was determined by the LC-MS method. Methyl tert-butyl ether (2 ml) was added to the reaction solution, and then water (2 ml) was added. The reaction solution was stirred until a solid precipitated, which was suction filtered. The filter cake was dried in vacuo to give a yellow solid, 4-(benzylamino)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (9 mg, yield: 22.3%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.62 (s, 1H), 8.79-8.82 (t, 1H), 8.60 (s, 1H) , 8.38-8.40 (d, 1H), 8.10-8.11 (d, 1H), 7.26-7.38 (m, 5H), 5.05-5.06 (d, 2H).

Molecular formula: C ₁₆ H ₁₂ N ₄ O, Molecular weight: 276.30, LC-MS (Pos, m/z)=277.02 [M+H ⁺ ].

Example 4

Synthesis of 4-((4-chlorobenzyl)amino)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 11)

4-Chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (30 mg, 0.146 mmol, 1.0 eq.) was dissolved in DMF (0.7 ml), after which ( 4-chlorophenyl)methylamine (29 mg, 0.204 mmol, 1.4 eq.) and DIPEA (75.4 mg, 0.584 mmol, 4 eq.). The system was heated to 100°C and left to react for 1.5 hours; the completion of the reaction was determined by the LC-MS method. After MTBE (2 ml) was added, the mixture was stirred and then water (2 ml) was added, stirred for 15 minutes, resulting in a solid precipitate which was suction filtered. The filter cake was washed with water and dried in vacuo to give a yellow solid (19 mg, yield: 41.9%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.56 (s, 1H), 8.80 (s, 1H), 8.60 (s, 1H), 8.38 -8.40 (d, 1H), 8.08-8.10 (d, 1H), 7.34-7.44 (t, 4H), 5.02 (s, 2H).

Molecular formula: C ₁₆ H ₁₁ ClN ₄ O, Molecular weight: 310.74 LC-MS (Pos, m/z)=309.1 [M+H ⁺ ].

Example 5

Synthesis of 2-oxo-4-(2-aza-spiro[3,5]nonan-2-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 13)

4-Chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.234 mmol, 1.0 eq.) was dissolved in DMF (1 ml), after which 2-aza was added -spiro[3,5]nonane (43 mg, 0.340 mmol, 1.4 eq.) and DIPEA (188 mg, 1.458 mmol, 4 eq.). The system was heated to 80°C and left to react for 2 hours; the completion of the reaction was determined by the LC-MS method. The reaction solution was cooled and filtered with suction. The filter cake was washed with water and dried in vacuo to give an off-white solid (27.42 mg, yield: 39.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.49 (s, 1H), 8.58 (s, 1H), 8.24-8.25 (d, 1H) , 7.74-7.76(d, 1H), 4.46(s, 1H), 1.69(s, 4H), 1.36-1.43(d, 6H).

Molecular formula: C ₁₇ H ₁₈ N ₄ O, Molecular weight: 294.36 LC-MS (Pos, m/z)=295.1 [M+H ⁺ ].

Example 6

Synthesis of 2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 16)

The starting material, 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.234 mmol, 1.0 eq.) was dissolved in DMF (1 ml), after which 7-aza-spiro[3,5]nonane hydrochloride (55 mg, 0.340 mmol, 1.4 eq.) and DIPEA (188 mg, 1.458 mmol, 6 eq.). The system was heated to 80°C and left to react for 2 hours; the completion of the reaction was determined by the LC-MS method. The reaction solution was cooled and filtered with suction. The filter cake was washed with water, suctioned off, and dried under reduced pressure to give a pale yellow solid, 2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro -1,7-naphthyridine-3-carbonitrile (23.24 mg, 33.7%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.98 (s, 1H), 8.64 (s, 1H), 8.32-8.34 (d, 1H) , 7.59-7.60 (d, 1H), 3.53 (s, 4H), 3.41-3.47 (m, 4H), 1.79-1.92 (m, 6H).

Molecular formula: C ₁₇ H ₁₈ N ₄ O, Molecular weight: 294.36 LC-MS (Pos, m/z)=295.1 [M+H ⁺ ].

Example 7

Synthesis of 4-((4-chlorophenyl)amino)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 18)

4-Chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (200 mg, 0.973 mmol, 1.0 eq.) was dissolved in DMF (3 ml), after which p-chloroaniline was added (174 mg, 1.362 mmol, 1.4 eq.) and DIPEA (752 mg, 3.893 mmol, 4 eq.). The system was heated to 80°C and left to react for 1.5 hours. The reaction solution was cooled, after which water (10 ml) was added and then extracted with EA. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was dissolved by adding a small amount of DCM and a methanol solution, and then a small amount of EA was added until a solid precipitated, which was filtered with suction. The filter cake was washed with a small amount of DCM and dried to give a yellow solid (44 mg, yield: 50.9%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.84 (s, 1H), 9.94 (s, 1H), 8.66 (s, 1H), 8.41 -8.42 (d, 1H), 8.10-8.12 (d, 1H), 7.47-7.49 (d, 2H), 7.35-7.37 (d, 2H).

Molecular formula: C ₁₅ H ₉ ClN ₄ O, Molecular weight: 296.71, LC-MS (Pos, m/z)=296.96 [M+H ⁺ ].

Example 8

Synthesis of (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 19)

Step 1: Synthesis of tert-butyl-(S)-3-hydroxypyrrolidine-1-carboxylate

(S)-pyrrolidin-3-ol (3.0 g, 24.28 mmol, 1.0 eq.) was dissolved in THF (54 ml), cooled to 0°C and then triethylamine (12.28 g , 121.38 mmol, 5 eq.). (Boc) ₂ O (5.83 g, 26.70 mmol, 1.1 eq.) was added dropwise to the mixture slowly with stirring. The system was slowly warmed to room temperature and allowed to react for 2 days. The reaction solution was concentrated under reduced pressure, after which water was added, stirred, and extracted with DCM three times. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a yellow oil, t-butyl-(S)-3-hydroxypyrrolidine-1-carboxylate (4.42 g, yield: 97.2%).

Step 2: Synthesis of tert-butyl-(S)-3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate

tert-Butyl-(S)-3-hydroxypyrrolidine-1-carboxylate (3.0 g, 16.02 mmol, 1 eq.) was placed in a reaction flask and dissolved by adding THF (30 ml). The system was cooled to 0° C. and then triethylamine (3.24 g, 32.04 mmol, 2 eq.) was added. Methansulfonyl chloride (2.2 g, 19.23 mmol, 1.2 eq.) was slowly added dropwise to the mixture. The mixture was slowly warmed to room temperature and left to react for 3 hours; completion of the reaction was determined by TLC (thin layer chromatography). The reaction solution was filtered with suction, and the filtrate was concentrated under reduced pressure to give a pale yellow oil, tert-butyl (S)-3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate, which was directly introduced into the next reaction step without purification .

Step 3: Synthesis of tert-butyl-(R)-3-(1H-pyrazol-1-yl)pyrrolidine-1-carboxylate

Pyrazole (1.2 g, 17.62 mmol, 1.1 eq.) was dissolved in DMAA (72 ml), cooled to 0°C, after which sodium hydride (2051 mg, 51.27 mmol, 3.2 eq. ., content 60%), and then the reaction was allowed to proceed for 1 hour under nitrogen atmosphere. tert-Butyl-(S)-3-((methylsulfonyl)oxy)pyrrolidine-1-carboxylate obtained in the above step was dissolved in a small amount of DMAA, and the resulting solution was slowly added to the reaction solution dropwise, heated to 100°C and left to react overnight under nitrogen. The reaction solution was cooled to room temperature, diluted with water, stirred, and extracted with EA. After separation of the liquids, the organic phases were combined, washed with water, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=20:1) to give a colorless oil, tert-butyl-(R)-3-(1H-pyrazol-1-yl)pyrrolidine-1-carboxylate (1.83 g , yield after two steps of synthesis: 48.2%).

Step 4: Synthesis of (R)-1-(pyrrolidin-3-yl)-1H-pyrazole

tert-Butyl-(R)-3-(1H-pyrazol-1-yl)pyrrolidine-1-carboxylate (200 mg, 0.8428 mmol, 1.0 eq.) was dissolved in DCM (4 ml), cooled to 0 °C, then trifluoroacetic acid (2 ml) was added dropwise, after which the reaction was left to proceed for 2-3 hours; the completion of the reaction was determined by TLC. The reaction system was then concentrated under reduced pressure, and made basic by adding an appropriate amount of DIPEA to give the crude product, (R)-1-(pyrrolidin-3-yl)-1H-pyrazole, which was used directly in the next step.

Step 5: Synthesis of (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4-Chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (124 mg, 0.602 mmol, 1.0 eq.) was dissolved in DMAA (2 ml), after which the crude product was added, (R)-1-(pyrrolidin-3-yl)-1H-pyrazole obtained in the above step and DIPEA (467 mg, 3.612 mmol, 6 eq.). The system was heated to 80°C and left to react for 1.5 hours; the completion of the reaction was determined by the LC-MS method. The reaction solution was diluted with water (10 ml) and extracted with EA (50 ml×3). The organic phases were combined, washed with water (30 ml), dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure to give a brownish yellow solid. The solid was suspended by adding a small amount of DCM and EA and filtered with suction. The filter cake was washed with a small amount of DCM and dried in vacuo to give a brownish yellow solid, (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-2-oxo-1, 2-dihydro-1,7-naphthyridine-3-carbonitrile (35.96 mg, yield from two synthetic steps: 13.9%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.64 (s, 1H), 8.61 (s, 1H), 8.26-8.28 (d, 1H) , 7.97-7.98 (d, 1H), 7.89-7.90 (d, 1H), 7.50-7.51 (d, 1H), 6.30 (s, 1H), 5 .14-5.20 (m, 1H), 4.51-4.56 (q, 1H), 4.32-4.36 (q, 1H), 4.17-4.30 (m, 2H) , 2.43-2.49 (m, 2H).

Molecular formula: C ₁₆ H ₁₄ N ₆ O, Molecular weight: 306.33, LC-MS (Pos, m/z)=307.0 [M+H ⁺ ].

Example 9

Synthesis of 2-oxo-4-(phenylamino)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 21)

4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50.0 mg, 0.243 mmol, 1.0 eq) and aniline (31.7 mg, 0.340 mmol, 1 .4 eq.) was dissolved in N,N-dimethylformamide (1.0 ml), N,N-diisopropylethylamine (125.8 mg, 0.973 mmol, 4.0 eq.) was added and heated under reflux to 80 ° C in within 2 hours. The completion of the reaction was determined by TLC. The system was cooled to room temperature and filtered with suction. The filter cake was washed with water (4.0 ml) for 30 minutes, suction filtered and dried at 60° C. to give a yellow solid (10.0 mg, yield: 20%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.80 (s, 1H), 9.93 (s, 1H), 8.67 (s, 1H), 8.42 -8.41 (m, 1H), 8.15-8.14 (m, 1H), 7.46-7.42 (m, 2H), 7.35-7.29 (m, 3H).

Molecular formula: C ₁₅ H ₁₀ N ₄ O, Molecular weight: 262.27, LC-MS (Pos, m/z)=263.59 [M+H ⁺ ].

Example 10

Synthesis of (R)-4-(3-(1Н-pyrazol-1-yl)pyrrolidin-1-yl)-6-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile ( Connections 23)

Step 1: Synthesis of (R)-1-(pyrrolidin-3-yl)-1H-pyrazole hydrochloride

Tert-butyl-(R)-3-(1H-pyrazol-1-yl)pyrrolidine-1-carboxylate (13.8 mg, 0.0583 mmol, 1.0 eq.) was dissolved in ethanol (1.0 ml) and then a solution of hydrogen chloride in ethanol (25%, 1.0 ml) was added at 0°C. The system was slowly warmed to room temperature and stirred for 1 hour; the completion of the reaction was determined by TLC. The reaction solution was concentrated, and the crude product was introduced into the next step.

Step 2: Synthesis of (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3 -carbonitrile

The crude product obtained as above was dissolved in N,N-dimethylformamide (1.0 ml), after which N,N-diisopropylethylamine (32.3 mg, 0.250 mmol, 6.0 eq.) and 4,6-dichloro- 2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (10.0 mg, 0.0416 mmol, 1.0 eq.). The mixture was heated under reflux to 80°C for 2 hours; the completion of the reaction was determined by TLC. The reaction mixture was concentrated under reduced pressure. The crude product was successively purified by thin layer chromatography (DCM:MeOH=20:1) and silica gel column chromatography (DCM:MeOH=100:1-40:1) to give a yellow solid (6.49 mg, yield: 64.9% ).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.77 (s, 1H), 8.41 (s, 1H), 8.00 (m, 1H), 7.90 -7.89 (m, 1H), 7.52-7.51 (m, 1H), 6.31-6.30 (m, 1H), 5.18-5.15 (m, 1H), 4 .54-4.50 (m, 1H), 4.36-4.16 (m, 3H), 2.01-1.99 (m, 1H), 0.88 (m, 1H).

Molecular formula: C ₁₆ H ₁₃ N ₆ O, Molecular weight: 340.77, LC-MS (Pos, m/z)=341.0 [M+H ⁺ ].

Example 11

Synthesis of 2-oxo-4-(2-oxa-7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 24)

Step 1: Synthesis of 2-oxo-4-(2-oxa-7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50.0 mg, 0.243 mmol, 1.0 eq) and 2-oxa-7-aza-spiro[ 3,5]nonane (58.6 mg, 0.170 mmol, 0.7 eq.) was dissolved in N,N-dimethylformamide (1.0 ml) and then N,N-diisopropylethylamine (188.4 mg, 1.458 mmol, 6.0 eq.). The system was heated to 80°C under reflux for 2 hours, and the completion of the reaction was determined by TLC. The system was cooled to room temperature and filtered with suction. The filter cake was washed with water (4.0 ml) for 30 minutes, suction filtered and dried at 60° C. to give a yellow solid (10.0 mg, yield: 20%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.02 (s, 1H), 8.66 (s, 1H), 8.35-8.34 (m, 1H) , 7.59-7.58 (m, 1H), 4.42 (s, 4H), 3.56-3.54 (m, 4H), 2.07-2.05 (m, 4H).

Molecular formula: C ₁₆ H ₁₆ N ₄ O ₂ , Molecular weight: 296.33, LC-MS (Pos, m/z)=297.61 [M+H ⁺ ].

Example 12

Synthesis of 2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compounds 25)

4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50.0 mg, 0.243 mmol, 1.0 eq.) and 6-aza-spiro[2.5]octane hydrochloride (50.2 mg, 0.340 mmol, 1.4 eq.) was dissolved in N,N-dimethylformamide (1.0 ml) and N,N-diisopropylethylamine (188.4 mg, 1.458 mmol, 6.0 eq. ). The system was heated to 80°C under reflux for 2 hours; the completion of the reaction was determined by TLC. The reaction mixture was cooled to room temperature and filtered with suction. The filter cake was washed with water (4.0 ml) for 30 minutes, suction filtered and dried at 60° C. to give a yellow solid (18.0 mg, yield: 36%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.02 (s, 1H), 8.66 (s, 1H), 8.36-8.34 (m, 1H) , 7.65-7.64 (m, 1H), 3.66-3.63 (m, 4H), 1.62 (m, 4H), 0.44 (s, 4H).

Molecular formula: C ₁₆ H ₁₆ N ₄ O, Molecular weight: 280.33, LC-MS (Pos, m/z)=281.1 [M+H ⁺ ].

Example 13

Synthesis of 4-(4-ethylphenyl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compounds 26)

4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (300.0 mg, 1.459 mmol, 1.0 eq.), (4-ethylphenyl)boric acid (262.6 mg, 1.751 mmol, 1.2 eq.), potassium phosphate (619.4 mg, 2.918 mmol, 2.0 eq.) and 1,1'-bis-diphenylphosphinoferrocene palladium dichloride (47.5 mg, 0.0729 mmol, 0.05 eq.) was dissolved in dioxane (9.0 ml) and water (4.5 ml). The system was heated to 115° C. under reflux overnight under nitrogen, and the completion of the reaction was determined by TLC. The reaction mixture was filtered with suction, and the filtrate was extracted with ethyl acetate (10.0 ml×3), dried with anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=200:1-40:1) to give a yellow solid (100.0 mg, yield: 33.3%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.83 (s, 1H), 8.80 (s, 1H), 8.38-8.36 (m, 1H) , 7.49 (m, 1H), 7.16-7.15 (m, 1H), 2.77-2.75 (m, 2H), 1.30-1.26 (m, 3H).

Molecular formula: C ₁₇ H ₁₃ N ₃ O, Molecular weight: 275.31, LC-MS (Pos, m/z)=276.1 [M+H ⁺ ].

Example 14

Synthesis of 4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compounds 27)

Step 1: Synthesis of 4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4,6-dichloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (180 mg, 0.75 mmol, 1.0 eq) and 1-(pyrrolidin-3-yl) -1H-pyrazole (303 mg, 1.28 mmol, 1.7 eq., 57.6%) was dissolved in DMF (5 ml) and then DIPEA (387 mg, 3.0 mmol, 4.0 eq.) was added. . The system was stirred at 80°C for 2 hours and cooled. The reaction solution was purified by reverse phase column chromatography (acetonitrile: water: ammonia solution=30:100:0.05%) to give a yellow solid (200 mg, yield: 78.4%).

Step 2: Synthesis of 4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-methyl-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (112.0 mg, 0.33 mmol, 1.0 eq.), trimethylcyclotriboroxane (330.0 mg, 1.30 mmol, 4.0 eq., 50%), cesium carbonate (322.0 mg, 0.99 mmol, 3.0 eq.), potassium phosphate (70.0 mg, 0.33 mmol, 1.0 eq.) and Pd(dppf) ₂ Cl ₂ (120 mg, 0.16 mmol, 0.5 eq.) were dissolved in 1, 4-dioxane (4 ml). The system was heated to 105° C. and stirred overnight under nitrogen. The completion of the reaction was determined by TLC. The reaction solution was quenched (ie, quickly stopped the reaction) by adding H ₂ O (10 ml), stirred for 30 minutes, cooled to room temperature and filtered with suction. The filter cake was washed with a small amount of ethyl acetate. The filtrate was extracted with ethyl acetate (10.0 ml×3). After separation of the liquids, the organic phase was washed with brine (10.0 ml×2), dried with anhydrous Na ₂ SO ₄ , filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=80:1-60:1) to give a yellow solid (55.0 mg, yield: 52.1%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.04 (s, 1H), 8.69 (s, 1H), 7.57 (m, 3H), 6.34 (s, 1H), 5.11 (s, 1H), 4.49-4.64 (m, 3H), 4.49 (s, 1H), 2.60-2.67 (m, 5H).

Molecular formula: C ₁₇ H ₁₆ N ₆ O, Molecular weight: 320.14, LC-MS (Neg, m/z)=319.17 [M-H] ^- .

Example 15

Synthesis of 4-(4-(2-hydroxyethyl)piperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 28)

4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (60 mg, 0.29 mmol, 1.0 eq.) was dissolved in N,N-dimethylformamide (1 ml). Then N,N-diisopropylethylamine (112 mg, 0.87 mmol, 3.0 eq) and 2-(piperidin-4-yl)ethan-1-ol (38 mg, 0.29 mmol, 1.0 eq) were added. .). The system was heated to 80°C and left to react for 2 hours, cooled to room temperature, resulting in a solid precipitate, and then filtered. The filter cake was washed successively with tetrahydrofuran (1 ml) and petroleum ether (1 ml) and dried at 45° C. to give a yellow solid (16 mg, yield: 18%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.98 (s, 1H), 8.65 (s, 1H), 8.33-8.35 (d, 1H) , 7.58-7.80 (d, 1H), 4.40-4.42 (m, 1H), 3.83-3.86 (m, 2H), 3.48-3.53 (m, 2H), 3.37-3.43(m, 2H), 1.84-1.87(m, 2H), 1.74-1.80(m, 1H), 1.45-1.50( m, 4H).

Molecular formula: C ₁₆ H ₁₈ N ₄ O ₂ , Molecular weight: 298.35, LC-MS (Neg, m/z)=297.15 [M-H] ^- .

Example 16

Synthesis of 6-(methylthio)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 30)

Step 1: Synthesis of 2-bromo-5-nitroisonicotinic acid

2-bromo-4-methyl-5-nitropyridine (2.5 g, 11.58 mmol) was dissolved in concentrated sulfuric acid (25 ml). The system was cooled to 0°C in an ice bath, after which chromium trioxide (3.88 g, 38.8 mmol) was added, then slowly warmed to room temperature and stirred overnight. The reaction solution was poured into ice water (75 ml), stirred for 10 minutes, and filtered with suction to give a white solid (2.5 g, yield: 87.8%).

Step 2: Synthesis of 2-(methylthio)-5-nitroisonicotinic acid

2-bromo-5-nitroisonicotinic acid (1.5 g, 6.1 mmol) was dissolved in DMF (30 ml) and the system was cooled to 0°C in an ice bath, after which sodium thiomethylate (1.07 g, 15 .25 mmol), then slowly warmed to room temperature and stirred overnight; the completion of the reaction was determined by the LC-MS method. The reaction solution was concentrated under reduced pressure, poured into water (8 ml), adjusted to pH 2 with 1 mol/l hydrochloric acid until a solid formed, and then suction filtered to give a red solid (1.0 g crude).

Step 3: Synthesis of 5-amino-2-(methylthio)isonicotinic acid

2-(methylthio)-5-nitroisonicotinic acid (1.0 g crude product) was dissolved in ethanol (10 ml) and saturated aqueous ammonium chloride solution (5 ml) and iron powder (5.23 g, 93.5 mmol) were added . The system was heated to 70°C and left to react overnight; the completion of the reaction was determined by the LC-MS method. The mixture was filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase column chromatography to give the product (400 mg, yield after two synthetic steps: 35.6%).

Step 4: Synthesis of 6-(methylthio)-2H-pyrido[3,4-d][1,3]oxazine-2,4(1H)-dione

5-Amino-2-(methylthio)isonicotinic acid (300.0 mg, 1.63 mmol) was dissolved in DMF (3 ml) and cooled to 0° C. in an ice bath. CDI (449.4 mg, 2.77 mmol) was added. The system was slowly warmed to room temperature and stirred overnight; the completion of the reaction was determined by TLC. The product was obtained by concentration under reduced pressure and was directly introduced into the next step without purification.

Step 5: Synthesis of 4-hydroxy-6-methylthio-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-(methylthio)-2H-pyrido[3,4-d][1,3]oxazine-2,4(1H)-dione (300.0 mg crude product) was dissolved in DMF (3 ml). Ethyl cyanoacetate (193.6 mg, 1.71 mmol) and triethylamine (329.9 mg, 3.26 mmol) were added and the system was stirred at 150° C. overnight; the completion of the reaction was determined by TLC. The mixture was concentrated under reduced pressure, after which water (10 ml) was added and then adjusted to pH 1 with hydrochloric acid until a solid precipitated, which was filtered with suction to give the product (140.0 mg of crude product).

Step 6: Synthesis of 2,4-dichloro-6-methylthio-1,7-naphthyridine-3-carbonitrile

4-Hydroxy-6-methylthio-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (130.0 mg of crude product) was dissolved in phosphorus oxychloride (2 ml), after which phosphorus pentachloride ( 168.7 mg, 1.1 mmol) and then heated to 100° C. under reflux overnight. The completion of the reaction was determined by TLC. The reaction solution was poured into ice water (15 ml), adjusted to pH 1 by adding saturated sodium bicarbonate aqueous solution, and filtered with suction. The filtrate was extracted with ethyl acetate and concentrated. The crude product was combined with the filter cake and purified by silica gel column chromatography (PE:EA=20:1) to give a yellow solid (60.0 mg, yield after three steps: 13.7%).

Step 7: Synthesis of 4-chloro-6-methylthio-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

2,4-dichloro-6-methylthio-1,7-naphthyridine-3-carbonitrile (50.0 mg, 0.186 mmol) was dissolved in TFA (4 ml) and water (1 ml). The system was heated at 100°C for 2 hours, and the completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure to give a product (68.0 mg of crude product) which was directly used in the next step without purification.

Step 8: Synthesis of 6-methylthio-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4-Chloro-6-methylthio-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (60.0 mg of crude product) was dissolved in DMF (2 ml). 7-Aza-spiro[3,5]nonane hydrochloride (54.12 mg, 0.336 mmol) and DIPEA (93.53 mg, 0.72 mmol) were added. The mixture was heated at 70°C for 2 hours, and the completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure, after which water and ethyl acetate were added, and the liquid was separated. The aqueous phase was extracted with ethyl acetate. The organic phase was combined, washed with water and brine, dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=80:1, 60:1, 40:1) to give the product (50.0 mg, yield after two synthetic steps: 79%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.94 (s, 1H), 8.54 (s, 1H), 7.33 (s, 1H), 3.50 (s, 4H), 2.55 (s, 2H), 1.92-1.80 (m, 10H).

Molecular formula: C ₁₈ H ₂₀ N ₄ OS, Molecular weight: 340.45, LC-MS (Pos, m/z)=341.0 [M+H] ⁺ .

Example 17

Synthesis of 6-(methylsulfonyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 31)

Step 1: Synthesis of 6-(methylsulfonyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-methylthio-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (40.0 mg, 0 .12 mmol) was dissolved in dichloromethane (8 ml). The mixture was cooled to 0°C, after which meta-chloroperoxybenzoic acid (59.2 mg, 0.24 mmol) was added, and then slowly warmed to room temperature to start the reaction. The completion of the reaction was determined by TLC and LC-MS. Saturated aqueous sodium bicarbonate solution (10 ml) and dichloromethane (10 ml) were added. After separation of the liquids, the aqueous phase was extracted with dichloromethane, the organic phases were combined, washed with water, then with brine, dried with anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1, 80:1) to give a yellow solid (10.0 mg, yield: 20.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.44 (s, 1H), 8.75 (s, 1H), 8.19 (s, 1H), 3.57 (s, 4H), 3.26 (s, 3H), 1.94-1.82 (m, 10H).

Molecular formula: C ₁₈ H ₂₀ N ₄ O ₃ S, Molecular weight: 372.44, LC-MS (Pos, m/z)=373.1 [M+H] ⁺ .

Example 18

Synthesis of 6-(1,2-dihydroxyethyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 32)

2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (700.2 mg, 2.285 mmol, 1.0 eq.) was dissolved in a mixture of tert-butanol (30 ml) and water (30 ml). The system was cooled to 0°C in an ice bath, after which methanesulfonamide (217.4 mg, 2.285 mmol, 1.0 eq.) and AD-mix-β (7.76 g) were added, and then vigorously stirred at room temperature in during the night. The completion of the reaction was determined by TLC. The reaction solution was filtered with suction, and the filter cake was washed with water (30 ml×3), dried in vacuo to give a yellow solid (504.7 mg, yield: 64.9%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.97 (s, 1H), 8.60 (s, 1H), 7.77 (s, 1H), 5.56 (s, 1H), 4.69-4.65 (d, 2H), 3.65 (s, 5H), 3.50 (s, 1H), 1.63 (s, 4H), 0.45 ( s, 4H).

Molecular formula: C ₁₈ H ₂₀ N ₄ O ₃ , Molecular weight: 340.38, LC-MS (Pos, m/z)=341.08 [M+H] ⁺ .

Example 19

Synthesis of 6-(1,2-dihydroxyethyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 33)

Step 1: Synthesis of 2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-chloro-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (4.61 g, 14 .02 mmol, 1.0 eq.), potassium trifluorovinyl borate (3.05 g, 22.77 mmol, 3.0 eq.), cesium carbonate (13.70 g, 42.06 mmol, 3.0 eq.) , Pd(PPh ₃ ) ₄ (810.1 mg, 0.70 mmol, 0.05 eq.) and Pd(dppf)Cl ₂ (512.9 mg, 0.70 mmol, 0.05 eq.) were dissolved in mixtures of 1,4-dioxane (40 ml) and water (10 ml). The system was heated to 105° C. and stirred overnight under nitrogen. The completion of the reaction was determined by TLC. H ₂ O (20 ml) was added to the reaction solution, and the system was stirred for 30 minutes, cooled to room temperature and filtered with suction. The filter cake was washed with a small amount of dichloromethane and the filtrate was extracted with dichloromethane (20 ml×3). The organic phases were combined, washed with brine (20 ml×2), dried with anhydrous Na ₂ SO ₄ , filtered with suction and concentrated under reduced pressure. The crude product was first purified by silica gel column chromatography (DCM:MeOH=80:1-30:1) and then washed with ethyl acetate (10 ml) to give a yellow solid (2.3 g, yield: 51.27%).

Step 2: Synthesis of 6-(1,2-dihydroxyethyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridin- 3-carbonitrile

2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (2.3 g, 7 .2 mmol, 1.0 eq.) was dissolved in a mixture of tert-butanol (60 ml) and water (60 ml). The system was cooled to 0°C in an ice bath, then methanesulfonamide (682.0 mg, 7.2 mmol, 1.0 eq.) and AD-mix-β (10 g) were slowly added, followed by vigorous stirring at room temperature in during the night. The completion of the reaction was determined by TLC. The reaction solution was filtered with suction, the filter cake was washed with water (30 ml×3) and dried in vacuo to give a yellow solid (857.0 mg, yield: 33.6%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.89 (s, 1H), 8.58 (s, 1H), 7.77 (s, 1H), 5.56 (s, 1H), 4.68 (m, 2H), 3.73 (s, 1H), 3.57 (m, 4H), 1.84-2.01 (m, 10H).

Molecular formula: C ₁₉ H ₂₂ N ₄ O ₃ , Molecular weight: 354.17, LC-MS (Pos, m/z)=355.11 [M+H] ⁺ .

Example 20

Synthesis of 4-(4-ethylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compounds 34)

Step 1: Synthesis of tert-butyl 4-ethyl-4-hydroxypiperidine-1-carboxylate

tert-Butyl-4-oxo-piperidine-1-carboxylate (2.0 g, 038 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (10.0 ml), then the solution was added slowly at 0°C under nitrogen atmosphere ethyl magnesium chloride (10.04 ml, 20.075 mmol, 2.0 eq.) diluted with THF (10.0 ml). The mixture was slowly warmed to room temperature and left to react overnight. After TLC analysis showed completion of the reaction, the crude product was purified by silica gel column chromatography (PE:EA=10:1 to 1:1) to give the product (800.0 mg, yield: 40%).

Step 2: Synthesis of tert-butyl-4-ethyl-3,6-dihydropyridine-1(2H)-carboxylate

tert-Butyl-4-ethyl-4-hydroxypiperidine-1-carboxylate (800.0 mg, 3.488 mmol, 1.0 eq.) was dissolved in dichloromethane (5.0 ml) and then pyridine (827.9 mg, 10.466 mmol, 3.0 eq.). Dichlorosulfoxide (830.1 mg, 6.977 mmol, 2.0 eq.) was added slowly to the mixture at 0° C. under a nitrogen atmosphere. The mixture was slowly warmed to room temperature and left to react overnight. After TLC analysis showed completion of the reaction, the reaction solution was poured into cold water and extracted with ethyl acetate. The organic phase was washed twice with hydrochloric acid (1 mol/l), dried over anhydrous Na ₂ SO ₄ and concentrated. The crude product was purified by silica gel column chromatography (PE:EA=20:1-5:1) to give the product as a yellow oil (450.0 mg, yield: 56%).

Step 3: Synthesis of tert-butyl-4-ethylpiperidine-1-carboxylate

tert-Butyl-4-ethyl-3,6-dihydropyridine-1(2H)-carboxylate (450.0 mg, 2.129 mmol, 1.0 eq.) was dissolved in methanol (10.0 ml) and then Pd/C was added (225.0 mg). The air in the system was replaced with hydrogen, and the reaction was carried out for 4 days. After TLC analysis showed completion of the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was directly introduced into the next step.

Step 4: Synthesis of 4-ethylpiperidine hydrochloride

The crude product obtained as described above was dissolved in ethanol (3.0 ml), and hydrogen chloride in ethanol (25%, 3.0 ml) was added dropwise to the reaction solution. The reaction in the resulting mixture was left to proceed at room temperature for 2 hours. After TLC analysis showed completion of the reaction, the reaction solution was concentrated under reduced pressure, and the crude product (102.0 mg) was directly introduced into the next step.

Step 5: Synthesis of 4-(4-ethylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

The crude product from the previous step (102.0 mg, 0.681 mmol, 1.0 eq.) was dissolved in N,N-dimethylformamide (3.0 ml) and N,N-diisopropylethylamine (528.1 mg, 4.086 mmol) was added , 6.0 eq.) and 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (140.0 mg, 0.681 mmol, 1.0 eq.). The reaction in the resulting system was left to proceed at 80°C for 2 hours. After TLC analysis showed completion of the reaction, the mixture was concentrated under reduced pressure, suspended and washed with ethyl acetate (1.5 ml) and methyl tert-butyl ether (5.0 ml) and filtered with suction. The filter cake was washed with water to give a yellow solid (100.0 mg, yield: 52.1%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.00 (s, 1H), 8.65 (s, 1H), 8.35-8.34 (m, 1H) , 7.60-7.58 (m, 1H), 3.87-3.84 (m, 2H), 3.42-3.39 (m, 2H), 1.88-1.85 (m, 2H), 1.44-1.36 (m, 5H), 1.01-0.89 (m, 3H).

Molecular formula: C ₁₆ H ₁₈ N ₄ O, Molecular weight: 282.35, LC-MS (Pos, m/z)=283.2 [M+H] ⁺ .

Example 21

Synthesis of 4-(4,4-bis(hydroxymethyl)piperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (compound 38)

4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (200.0 mg, 0.973 mmol, 1.0 eq.), piperidine-4,4-diyldimethanol (265.1 mg, 1.459 mmol, 1.5 eq.) and DIPEA (1005.8 mg, 7.782 mmol, 8.0 eq.) were dissolved in DMF (5 ml). The mixture was stirred at 80° C. and left to react for 3 hours. The completion of the reaction was determined by TLC. The mixture was concentrated under reduced pressure, washed for 3 hours with the addition of water (10 ml) and ethyl acetate (10 ml) and filtered with suction. The filter cake was rinsed with a small amount of methanol to give a dark red solid (55.0 mg, yield: 18.0%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.94 (s, 1H), 8.65 (s, 1H), 8.36-8.34 (d, 1H) , 7.60-7.59 (d, 1H), 4.55 (t, 2H), 3.63 (t, 4H), 3.43-3.42 (d, 4H), 1.65 (t , 4H).

Molecular formula: C ₁₆ H ₁₈ N ₄ O ₃ , Molecular weight: 314.35, LC-MS (Pos, m/z)=315.04 [M+H] ⁺ .

Example 22

Synthesis of (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-morpholino-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile ( Compound 39)

(R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (60 .0 mg, 0.176 mmol), morpholine (36.85 mg, 0.423 mmol), Pd ₂ (dba) ₃ (6.7 mg, 0.007 mmol), sodium tert-butoxide (47.54 mg, 0.493 mmol) and XPhos (6.7 mg, 0.014 mmol) was dissolved in 1,4-dioxane (1 ml) and DMA (0.2 ml). The reaction in the resulting system was left to proceed under microwave heating to 110°C for 4 hours. The completion of the reaction was determined by TLC. The mixture was concentrated under reduced pressure, after which water was added and then extracted with ethyl acetate. The organic phases were combined, washed with water then brine, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=80:1, 60:1, 40:1, 20:1) to give a red solid (13.0 mg, 18.9%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.30 (s, 1H), 8.30 (s, 1H), 7.89-7.88 (d, 1H) , 7.50 (d, J=1.2 Hz, 1H), 7.15 (d, 1H), 6.30-6.29 (m, 1H), 5.17-5.15 (m, 1H ), 4.52-4.48(m, 1H), 4.36-4.32(m, 1H), 4.29-4.16(m, 2H), 3.74-3.72(m , 4H), 3.37 (s, 3H), 2.46-2.33 (m, 2H).

Molecular formula: C ₂₀ H ₂₁ N ₇ O ₂ , Molecular weight: 391.44, LC-MS (Pos, m/z)=392.1 [M+H] ⁺ .

Example 23

Synthesis of 4-(2-hydroxy-7-aza-spiro[3,5]nonan-7-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (compound 40)

4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.24 mmol, 1.0 eq.) was dissolved in DMF and 7-aza-spiro[ hydrochloride was added 3,5]nonan-2-ol (60.5 mg, 0.34 mmol, 1.4 eq.) and DIPEA (188.5 mg, 1.46 mmol, 6.0 eq.). The reaction in the resulting system was left to proceed at 80°C for 2 hours. The completion of the reaction was determined by LC-MS. MTBE (2 ml) and H ₂ O (2 ml) were added. The mixture was vibrated and suction filtered to give the product (37 mg, yield: 49.7%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.97 (s, 1H), 8.64 (s, 1H), 8.32-8.33 (m, 1H) , 7.57-7.59 (m, 1H), 4.97-4.98 (m, 1H), 4.12-4.19 (m, 1H), 3.50-3.55 (m, 4H), 2.21-2.26 (m, 2H), 1.74-1.75 (m, 4H), 1.64-1.69 (m, 2H).

Molecular formula: C ₁₇ H ₁₈ N ₄ O ₂ , Molecular weight: 310.36, LC-MS (Pos, m/z)=311.02 [M+H] ⁺ .

Example 24

Synthesis of 4-(2-cyano-7-aza-spiro[3,5]nonan-7-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (compound 42)

4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.24 mmol, 1.0 eq.) was dissolved in DMF and 7-aza-spiro[ hydrochloride was added 3,5]nonane-2-carbonitrile (51.2 mg, 0.34 mmol, 1.4 eq.) and DIPEA (188.5 mg, 1.46 mmol, 6.0 eq.). The reaction in the resulting system was left to proceed at 80° C. for 1.5 hours, and the completion of the reaction was determined by the LC-MS method. The mixture was filtered with suction to give the product (19 mg, yield: 24.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.00 (s, 1H), 8.64 (s, 1H), 8.32-8.34 (m, 1H) , 7.57-7.58 (m, 1H), 3.52-3.55 (m, 4H), 3.38-3.46 (m, 1H), 2.29-2.34 (m, 2H), 2.15-2.20 (m, 2H), 1.84-1.88 (m, 4H).

Molecular formula: C ₁₈ H ₁₇ N ₅ O, Molecular weight: 319.37, LC-MS (Pos, m/z)=320.05 [M+H] ⁺ .

Example 25

Synthesis of 4-(2,2-difluoro-7-aza-spiro[3,5]nonan-7-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 43)

Step 1: Synthesis of tert-butyl 2,2-difluoro-7-aza-spiro[3,5]nonane-7-carboxylate

tert-Butyl 2-oxo-7-aza-spiro[3,5]nonane-7-carboxylate (500.0 mg, 2.09 mmol) was dissolved in dichloromethane (4 ml). The system was cooled to 0°C in an ice bath, after which DAST (673.8 mg, 4.18 mmol) was added, slowly warmed to room temperature and stirred overnight. The completion of the reaction was determined by TLC. The reaction solution was added to a saturated sodium bicarbonate aqueous solution. The aqueous phase was extracted with dichloromethane. The organic phases were combined, washed with water then brine and concentrated. The crude product was purified by silica gel column chromatography (PE:EA=30:1, 25:1, 20:1) to give the product (170.0 mg, yield: 31.2%).

Step 2: Synthesis of 2,2-difluoro-7-aza-spiro[3,5]nonane

tert-Butyl 2,2-difluoro-7-aza-spiro[3,5]nonane-7-carboxylate (170.0 mg, 0.65 mmol) was dissolved in dichloromethane (3 ml). The system was cooled to 0°C in an ice bath, after which trifluoroacetic acid (1.5 ml) was added, slowly warmed to room temperature and stirred for 2 hours. The completion of the reaction was determined by TLC. The mixture was concentrated under reduced pressure to give the product (115.0 mg crude).

Step 3: Synthesis of 4-(2,2-difluoro-7-aza-spiro[3,5]nonan-7-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (95.6 mg, 0.465 mmol) and 2,2-difluoro-7-aza-spiro[3.5]nonane (104.72 mg of crude product) was dissolved in DMF (3 ml) and DIPEA (181.2 mg, 1.395 mmol) was added. The mixture was heated to 80°C and left to react for 2 hours; the completion of the reaction was determined by TLC. The reaction solution was poured into water (10 ml), filtered with suction, and the filter cake was washed with ethyl acetate and petroleum ether to give the product (50.0 mg).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.03 (s, 1H), 8.65 (s, 1H), 8.35-8.33 (d, 1H) , 3.58 (s, 4H), 2.51 (s, 2H), 1.99-1.88 (d, 4H), 1.26-1.24 (d, 2H).

Molecular formula: C ₁₇ H ₁₆ F ₂ N ₄ O, Molecular weight: 330.34, LC-MS (Pos, m/z)=331.1 [M+H] ⁺ .

Example 26

Synthesis of 6-amino-2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 44)

6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (30 mg, 0.095 mmol, 1.0 eq.), LiHMDS in THF (1 mol/l, 1 ml), Pd ₂ (dba) ₃ (9 mg, 0.0095 mmol, 0.1 eq.) and 2-(dicyclohexylphosphino)biphenyl (7 mg, 0.0191 mmol, 0.2 eq.) were placed in a microwave tube and heated under microwave radiation to 100° C. for 2 hours. The reaction solution was cooled, after which 1 mol/L hydrochloric acid (12 ml) was added and stirred for 20 minutes. The system was brought to a slightly alkaline reaction by adding aqueous sodium carbonate solution and extracted with EA. The organic phase was dried with anhydrous sodium sulfate and separated twice on a preparative thin layer chromatography plate (DCM:MeOH=30:1.15:1) to give the product (1.03 mg, yield: 3.7%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.55 (s, 1H), 8.13 (s, 1H), 6.71 (s, 1H), 5.93 (s, 1H), 7.32-7.34 (t, 1H), 3.54-3.56 (t, 4H), 1.96-2.03 (m, 4H), 1.63 (m , 2H), 0.86 (m, 2H).

Molecular formula: C ₁₆ H ₁₇ N ₅ O, Molecular weight: 295.35, LC-MS (Pos, m/z)=296.05 [M+H] ⁺ .

Example 27

Synthesis of 6-methylamino-2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (compound 45)

6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.159 mmol, 1.0 eq.), methylamine in THF (1 mol/l, 0.8 ml, 0.794 mmol, 5 eq.), Pd ₂ (dba) ₃ (6 mg, 0.006 mmol, 0.04 eq.), tert sodium -butoxide (46 mg, 0.476 mmol, 2.8 eq.) and Xphos (6 mg, 0.013 mmol, 0.08 eq.) were dissolved in 1,4-dioxane (1 ml) and DMAA (0.2 ml) . The reaction solution was heated under microwave radiation to 110°C for 4 hours, cooled, after which water was added and extracted with EA. The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1-50:1) to give the product (5.69 mg, yield: 11.6%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.54 (s, 1H), 8.21 (s, 1H), 6.54-6.55 (d, 1H) , 6.61 (s, 1H), 3.57 (m, 4H), 2.77-2.78 (d, 3H), 1.62 (m, 4H), 0.43 (m, 4H).

Molecular formula: C ₁₇ H ₁₉ N ₅ O, Molecular weight: 309.37, LC-MS (Pos, m/z)=310.17 [M+H] ⁺ .

Example 28

Synthesis of 6-dimethylamino-2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 46)

6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.159 mmol, 1.0 eq.), dimethylamine in THF (2 mol/l, 0.4 ml, 0.794 mmol, 5 eq.), Pd ₂ (dba) ₃ (6 mg, 0.006 mmol, 0.04 eq.), tert sodium -butanol (43 mg, 0.445 mmol, 2.8 eq.) and Xphos (6 mg, 0.013 mmol, 0.08 eq.) were dissolved in 1,4-dioxane (1 ml) and DMAA (0.2 ml) . The reaction solution was heated under microwave radiation to 110°C for 4 hours, cooled, after which water was added and extracted with EA. The organic phase was dried with anhydrous sodium sulfate. The crude product was isolated by preparative thin layer chromatography (DCM:MeOH=20:1) to give the product (26.63 mg, yield: 51.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.58 (s, 1H), 8.34 (s, 1H), 6.63 (s, 1H), 3.62 (s, 4H), 3.05 (s, 6H), 1.62 (m, 4H), 0.44 (m, 4H).

Molecular formula: C ₁₈ H ₂₁ N ₅ O, Molecular weight: 323.40, LC-MS (Pos, m/z)=324.1 [M+H] ⁺ .

Example 29

Synthesis of 6-isopropyl-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 47)

6-chloro-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.152 mmol, 1.0 eq.), dimethylamine in THF (2 mol/l, 0.38 ml, 0.76 mmol, 5 eq.), Pd ₂ (dba) ₃ (5.7 mg, 0.006 mmol, 0.04 eq. .), sodium tert-butoxide (41.5 mg, 0.43 mmol, 2.8 eq.) and Xphos (5.72 mg, 0.012 mmol, 0.08 eq.) were dissolved in 1,4-dioxane (1 ml) and DMAA (0.2 ml). The reaction solution was heated under microwave radiation to 110°C for 4 hours, cooled, after which water was added and extracted with EA. The organic phase was dried with anhydrous sodium sulfate. The crude product was isolated by preparative thin layer chromatography (developing agent: DCM : MeOH = 20:1) to give 6-isopropyl-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1 ,2-dihydro-1,7-naphthyridine-3-carbonitrile (11.56 mg, yield: 22.5%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.57 (s, 1H), 8.30 (s, 1H), 6.59 (s, 1H), 3.51 (s, 4H), 3.03 (s, 6H), 1.79-2.07 (m, 10H).

Molecular formula: C ₁₉ H ₂₃ N ₅ O, Molecular weight: 337.43, LC-MS (Pos, m/z)=338.15 [M+H] ⁺ .

Example 30

Synthesis of 6-((dimethylamino)methyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile ( Connection 48)

6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (10 mg, 0.03 mmol, 1.0 eq.) was dissolved in 1,2-dichloroethane (1 ml) and then a solution of dimethylamine in tetrahydrofuran (2 mol/l, 0.06 ml, 0.13 mmol, 4.0 eq.) was added. The mixture was stirred at room temperature for 1 hour. While cooling with an ice bath, sodium triacetoxyborohydride (21 mg, 0.10 mmol, 3.0 eq.) was added. The system was stirred at room temperature for 2 hours and quenched by adding water (1 ml). The mixture was extracted with dichloromethane (5 ml×2). The organic phases were combined and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane:methanol=15:1) to give an off-white solid (1.6 mg, yield: 14.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.03 (s, 1H), 8.62 (s, 1H), 7.66 (s, 1H), 3.64 (m, 6H), 2.26 (m, 6H), 1.62 (m, 4H), 0.45 (m, 4H).

Molecular formula: C ₁₉ H ₂₃ N ₅ O, Molecular weight: 337.19, LC-MS (Pos, m/z)=336.17 [M+H] ⁺ .

Example 31

Synthesis of 6-(azetidin-1-yl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 49)

6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.159 mmol, 1.0 eq.), azetidine hydrochloride (74 mg, 0.794 mmol, 5 eq.), Pd ₂ (dba) ₃ (6 mg, 0.006 mmol, 0.04 eq.), sodium tert-butoxide (119 mg, 1.239 mmol, 7.8 eq.) and Xphos (6 mg, 0.013 mmol, 0.08 eq.) were dissolved in 1,4-dioxane (1 ml) and DMAA (0.2 ml). The reaction solution was heated under microwave radiation to 110°C for 4 hours, cooled, after which water was added and extracted with EA. The organic phase was dried with anhydrous sodium sulfate. The crude product was purified by preparative thin layer chromatography (DCM:MeOH=20:1) to give the product (1.25 mg, yield: 2.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.65 (s, 1H), 8.28 (s, 1H), 6.42 (s, 1H), 3.92 -3.95 (t, 4H), 3.59-3.62 (t, 4H), 2.29-2.36 (m, 2H), 1.60 (m, 4H), 0.432 (m, 4H ).

Molecular formula: C ₁₉ H ₂₁ N ₅ O, Molecular weight: 335.41, LC-MS (Pos, m/z)=336.15 [M+H] ⁺ .

Example 32

Synthesis of 6-(3-hydroxyazetidin-1-yl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridin-3 -carbonitrile (Compound 50)

6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (200 mg, 0.64 mmol, 1.0 eq.) and azetidin-3-ol hydrochloride (278 mg, 2.54 mmol, 4.0 eq.) were dissolved in 1,4-dioxane (5 ml). Sodium butoxide (478 mg, 4.96 mmol, 7.8 eq.), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (24 mg, 0.051 mmol, 0.08 eq.) and Pd ₂ ( dba) ₃ (24 mg, 0.025 mmol, 0.04 eq.). The mixture was heated to 120° C. under nitrogen and allowed to react for 3 hours. The completion of the reaction was determined by LC-MS. The reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH = 20:1), then suspended and washed with EA and suction filtered to give the product (74 mg, yield: 33%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.64 (s, 1H), 8.27 (s, 1H), 6.44 (s, 1H), 5.64 -5.66 (t, 1H), 4.56-4.59 (m, 1H), 4.14-4.18 (t, 2H), 3.60-3.66 (m, 6H), 1 .59 (m, 6H), 0.43 (m, 4H).

Molecular formula: C ₁₉ H ₂₁ N ₅ O ₂ , Molecular weight: 351.41, LC-MS (Pos, m/z)=352.17 [M+H] ⁺ .

Example 33

Synthesis of 6-(3-hydroxyazetidin-1-yl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridin-3 -carbonitrile (Compound 51)

6-chloro-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (80 mg, 0.243 mmol, 1.0 eq.) and azetidin-3-ol hydrochloride (89 mg, 0.81 mmol, 3.3 eq.) were dissolved in 1,4-dioxane (5 ml). Sodium butoxide (70 mg, 0.73 mmol, 3.0 eq.), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (11 mg, 0.024 mmol, 0.1 eq.) and Pd ₂ ( dba) ₃ (12 mg, 0.012 mmol, 0.05 eq.). The mixture was heated to 120° C. under nitrogen and allowed to react for 3 hours. The completion of the reaction was determined by LC-MS. The reaction solution was filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=10:1) to give a yellow solid (9 mg, yield: 10%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.59 (s, 1H), 8.27 (s, 1H), 6.41 (s, 1H), 5.63 -5.65 (d, 1H), 4.56-4.63 (m, 1H), 4.14-4.18 (m, 2H), 3.63-3.67 (m, 2H), 3 .49 (s, 4H), 1.74-1.94 (m, 10H).

Molecular formula: C ₂₀ H ₂₃ N ₅ O ₂ , Molecular weight: 365.44, LC-MS (Pos, m/z)=366.14 [M+H] ⁺ .

Example 34

Synthesis of 6-(3-aminoazetidin-1-yl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridin-3 -carbonitrile (Compound 52)

Step 1: Synthesis of tert-butyl-(1-(3-cyano-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7- naphthyridin-6-yl)azetidin-3-yl)carbamate

6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (300 mg, 0.95 mmol, 1.0 eq.) and tert-butylazetidin-3-ylcarbamate (795.5 mg, 3.81 mmol, 4.0 eq.) were dissolved in 1,4-dioxane, after which Pd ₂ (dba) ₃ was added (36.1 mg, 0.04 mmol, 0.04 eq.), t-BuONa (643.3 mg, 6.67 mmol, 7.0 eq.) and Xphos (36.3 mg, 0.08 mmol , 0.08 eq.). The reaction in the resulting mixture was left to proceed at 120°C for 2 hours. The completion of the reaction was determined by LC-MS. The mixture was concentrated, and the crude product was purified by silica gel column chromatography (DCM:MeOH=50:1-20:1) to give the product (177 mg, yield: 41.2%).

Step 2: Synthesis of 6-(3-aminoazetidin-1-yl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7- naphthyridine-3-carbonitrile

tert-Butyl-(1-(3-cyano-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-6- yl)azetidin-3-yl)carbamate (177 mg, 0.39 mmol, 1.0 eq.) was dissolved in DCM and trifluoroacetic acid (1 ml) was added. The reaction in the resulting mixture was left to proceed at 25°C for half an hour. The completion of the reaction was determined by LC-MS. The mixture was concentrated, and the crude product was purified by silica gel column chromatography (DCM:MeOH=10:1) to give the product (25.5 mg, yield: 18.7%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 8.29 (s, 1H), 6.46 (s, 1H), 3.94-4.18 (m, 3H) , 3.67-3.70 (m, 2H), 3.60 (m, 4H), 1.60 (s, 4H), 1.09-1.13 (m, 2H), 0.81-0 .85 (m, 4H).

Molecular formula: C ₂₀ H ₂₂ N ₆ O, Molecular weight: 350.43, LC-MS (Pos, m/z)=351.15 [M+H] ⁺ .

Example 35

Synthesis of 6-(3-aminoazetidin-1-yl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3 -carbonitrile (Compound 53)

Step 1: Synthesis of (1-(3-cyano-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-6- yl)azetidin-3-yl)carbamate

6-chloro-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (200 mg, 0.61 mmol, 1.0 eq.) and tert-butyl azetidin-3-yl carbamate (105 mg, 0.61 mmol, 1.0 eq.) were dissolved in 1,4-dioxane (5 ml), after which tert- sodium butoxide (175 mg, 1.82 mmol, 3.0 eq.), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (25 mg, 0.061 mmol, 0.1 eq.) and Pd ₂ (dba ) ₃ (30 mg, 0.031 mmol, 0.05 eq.). The temperature was raised to 120°C, and the reaction in the resulting system was left to proceed for 3 hours in a nitrogen atmosphere. The completion of the reaction was determined by the LC-MS method. The reaction solution was filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=20:1) to give a yellow solid (140 mg, yield: 49%).

Step 2: Synthesis of 6-(3-aminoazetidin-1-yl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7- naphthyridine-3-carbonitrile

(1-(3-cyano-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridin-6-yl)azetidine- 3-yl)carbamate (140 mg, 0.30 mmol, 1.0 eq.) was dissolved in dichloromethane (4 ml), after which trifluoroacetate (1 ml) was added and left to react for 2 hours at room temperature. The completion of the reaction was determined by the LC-MS method. A solid precipitate formed. The mixture was filtered and the filter cake was rinsed with dichloromethane (5 ml×2) and dried at 45° C. to give the product (19 mg, yield: 17%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 8.26 (s, 1H), 6.39 (s, 1H), 4.10-4.14 (m, 2H) , 3.79-3.85 (m, 1H), 3.50-3.53 (m, 6H), 1.79-1.93 (m, 10H).

Molecular formula: C ₂₀ H ₂₄ N ₆ O, Molecular weight: 364.45, LC-MS (Pos, m/z)=365.16 [M+H] ⁺ .

Example 36

Synthesis of 6-(azetidin-1-yl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 54)

6-chloro-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (150 mg, 0.46 mmol, 1.0 eq.) and azetidine (130 mg, 2.28 mmol, 5.0 eq.) were dissolved in tetrahydrofuran (5 ml) and DMAA (1 ml), after which sodium tert-butoxide (343 mg, 3.56 mmol, 7.8 eq.), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (17 mg, 0.036 mmol, 0.08 eq.) and Pd ₂ (dba) ₃ (17 mg, 0.018 mmol, 0.04 eq.). The temperature was raised to 110° C. and the resulting system was allowed to react for 8 hours under nitrogen. The completion of the reaction was determined by the LC-MS method. The reaction solution was poured into water and extracted with EA. The organic phase was dried and concentrated. The crude product was purified by preparative thin layer chromatography (DCM:MeOH=20:1) to give the product (61 mg, yield: 38%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.60 (s, 1H), 8.27 (s, 1H), 6.37 (s, 1H), 3.90 -3.94 (t, 4H), 3.48 (s, 4H), 2.30-2.36 (m, 2H), 1.78-1.92 (m, 10H).

Molecular formula: C ₂₀ H ₂₃ N ₅ O, Molecular weight: 349.44, LC-MS (Pos, m/z)=350.12 [M+H] ⁺ .

Example 37

Synthesis of 6-chloro-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 55)

4,6-dichloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (500.0 mg, 2.08 mmol, 1.0 eq.), 7-aza-spiro[ hydrochloride 3,5]nonane (505.1 mg, 3.12 mmol, 1.5 eq.) and DIPEA (2153.6 mg, 16.66 mmol, 8.0 eq.) were dissolved in DMF (10 ml), stirred for the reaction to proceed for 3 hours at 80°C. The completion of the reaction was determined by TLC. The system was concentrated under reduced pressure, after which water (10 ml) and ethyl acetate (10 ml) were added until a suspension formed and washed for 3 hours, then filtered with suction. The filter cake was rinsed with a small amount of methanol to give a dark red solid (67.0 mg, yield: 9.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.11 (s, 1H), 8.45 (s, 1H), 3.54 (s, 4H), 1.86 -1.80 (d, 10H).

Molecular formula: C ₁₇ H ₁₇ ClN ₄ O, Molecular weight: 328.80, LC-MS (Pos, m/z)=329.15 [M+H] ⁺ .

Example 38

Synthesis of 6-morpholino-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 56)

6-chloro-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50.0 mg, 0.152 mmol), morpholine (32.23 mg, 0.37 mmol), Pd ₂ (dba) ₃ (5.7 mg, 0.006 mmol), sodium tert-butoxide (41.5 mg, 0.43 mmol) and XPhos ( 5.72 mg, 0.012 mmol) was dissolved in 1,4-dioxane (1 ml) and DMA (0.2 ml). In a protective atmosphere of nitrogen, the reaction in the resulting system was left to proceed under microwave heating to 110°C for 4 hours. The completion of the reaction was determined by TLC. The system was concentrated under reduced pressure, after which water was added and extracted with ethyl acetate. The organic phases were combined, washed with water, washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=80:1-20:1) to give a product (15.0 mg).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.67 (s, 1H), 8.35 (s, 1H), 6.78 (d, 1H), 3.76 -3.74 (t, 4H), 3.38-3.35 (t, 4H), 1.99-1.80 (m, 10H).

Molecular formula: C ₂₁ H ₂₅ N ₅ O ₂ , Molecular weight: 379.46, LC-MS (Pos, m/z)=380.17 [M+H] ⁺ .

Example 39

Synthesis of 6-(4-methylpiperazin-1-yl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridin-3 -carbonitrile (Compound 57)

6-chloro-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (80.0 mg, 0.243 mmol, 1.0 eq.), N-methylpiperazine (58.4 mg, 0.584 mmol, 2.4 eq.), sodium tert-butoxide (65.7 mg, 0.681 mmol, 2.8 eq.), tris( dibenzylideneacetone) dipalladium (9.2 mg, 0.00972 mmol, 0.04 eq.) and 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (9.3 mg, 0.0194 mmol, 0.08 eq. .) was dissolved in 1,4-dioxane (2.0 ml) and N,N-dimethylacetamide (0.4 ml) and stirred at 120° C. overnight under nitrogen atmosphere. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure, after which water (5.0 ml) was added, extracted with ethyl acetate (5.0 ml), dried with anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=80:1-20:1) to give a yellow solid (20.0 mg, yield: 25%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.64 (s, 1H), 8.33 (s, 1H), 6.77 (s, 1H), 3.52 -3.32 (m, 6H), 2.47 (s, 3H), 2.24 (m, 2H), 1.91-1.78 (m, 8H), 1.36 (m, 2H), 1.34-1.24 (m, 4H).

Molecular formula: C ₂₂ H ₂₈ N ₆ O, Molecular weight: 392.51, LC-MS (Pos, m/z)=393.21 [M+H] ⁺ .

Example 40

Synthesis of 2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 58)

Step 1: Synthesis of 6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4,6-dichloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (4.00 g, 16.66 mmol, 1.0 eq.), 6-aza-spiro[2 ,5]octane (2.22 g, 20.00 mmol, 1.2 eq.) and DIPEA (8.61 g, 66.66 mmol, 4.0 eq.) were dissolved in DMF (40 ml), stirred in for 3 hours at 80°C. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure, after which water (30 ml) and ethyl acetate (30 ml) were added until a suspension formed, washed for 3 hours, and filtered with suction. The filter cake was rinsed with a small amount of methanol to give a yellow solid (3.63 g, yield: 69.2%).

Step 2: Synthesis of 2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (4.63 g, 14 .71 mmol, 1.0 eq.), potassium trifluorovinylborate (5.91 g, 44.13 mmol, 3.0 eq.), cesium carbonate (14.38 g, 44.13 mmol, 3.0 eq.) , Pd(PPh ₃ ) ₄ (849.9 mg, 0.74 mmol, 0.05 eq.) and Pd(dppf)Cl ₂ (538.1 mg, 0.74 mmol, 0.05 eq.) were dissolved in mixtures of 1,4-dioxane (40 ml) and water (10 ml). The temperature was raised to 105° C. and the system was stirred to allow the reaction to proceed overnight under a nitrogen atmosphere. The completion of the reaction was determined by TLC. H ₂ O (20 ml) was added. The reaction solution was stirred for 30 minutes, cooled to room temperature and filtered with suction, and the filter cake was washed with a small amount of dichloromethane. The filtrate was extracted with dichloromethane (20 ml×3). The organic phases were combined, washed with brine (20 ml×2), dried with anhydrous Na ₂ SO ₄ , filtered with suction and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=80:1-30:1) and washed with ethyl acetate (10 ml) to give a yellow solid (1.28 g, yield: 28.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.09 (s, 1H), 8.65 (s, 1H), 7.590 (s, 1H), 6.96-6 .89 (m, 1H), 6.17 (d, J=16 Hz, 1H), 5.40 (d, J=12 Hz, 1H), 3.68 (m, 4H), 1.64 (m , 4H), 0.44 (m, 4H).

Molecular formula: C ₁₈ H ₁₈ N ₄ O, Molecular weight: 306.15, LC-MS (Neg, m/z)=305.06 [M-H] ^- .

Example 41

Synthesis of 6-cyclopropyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 59)

6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.16 mmol, 1.0 eq.) and cyclopropylboronic acid (55 mg, 0.64 mmol, 4.0 eq.) were dissolved in 1,4-dioxane (3 ml), after which cesium carbonate (155 mg, 0.48 mmol, 3.0 eq.), potassium phosphate (34 mg, 0.16 mmol, 1.0 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (12 mg, 0.016 mmol, 0.0 1 equivalent). The temperature was raised to 120°C, and the reaction in the resulting system was left to proceed for 3 hours in a nitrogen atmosphere. The completion of the reaction was determined by the LC-MS method. The resulting mixture was filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=20:1) to give the product (2.5 mg, yield: 4.9%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.89 (s, 1H), 8.52 (s, 1H), 7.47 (s, 1H), 3.65 (m, 4H), 2.16-2.25 (m, 1H), 1.63 (s, 4H), 0.93-0.95 (m, 2H), 0.86-0.87 (m , 2H), 0.44 (m, 4H).

Molecular formula: C ₁₉ H ₂₀ N ₄ O, Molecular weight: 320.40, LC-MS (Pos, m/z)=321.92 [M+H] ⁺ .

Example 42

Synthesis of 6-cyclopropyl-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 60)

6-chloro-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.15 mmol, 1.0 eq.) and cyclopropylboronic acid (53 mg, 0.61 mmol, 4.0 eq.) were dissolved in 1,4-dioxane (3 ml), after which cesium carbonate (149 mg, 0.46 mmol, 3.0 eq.), potassium phosphate (33 mg, 0.15 mmol, 1.0 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (11 mg, 0.015 mmol, 0.0 1 equivalent). In a protective atmosphere of nitrogen, the reaction in the resulting system was left to proceed under microwave heating to 120°C for 3 hours. The completion of the reaction was determined by the LC-MS method. The reaction solution was filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=20:1) to give the product (4 mg, yield: 8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.83 (s, 1H), 8.51 (s, 1H), 7.43 (s, 1H), 3.54 (s, 4H), 2.23-2.25 (m, 1H), 2.17-2.22 (m, 10H), 0.93-0.95 (m, 2H), 0.87-0 .88 (m, 2H).

Molecular formula: C ₂₀ H ₂₂ N ₄ O, Molecular weight: 334.42, LC-MS (Pos, m/z)=335.07 [M+H] ⁺ .

Example 43

Synthesis of 6-((2-(dimethylamino)ethyl)(methyl)amino)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1, 7-naphthyridine-3-carbonitrile (Compound 61)

6-chloro-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.15 mmol, 1.0 eq.) and N ¹ ,N ¹ ,N ² -trimethylethane-1,2-diamine (78 mg, 0.76 mmol, 5.0 eq.) were dissolved in tetrahydrofuran (2 ml) and DMAA ( 0.2 ml), after which sodium tert-butoxide (114 mg, 1.19 mmol, 7.8 eq.), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (6 mg, 0.012 mmol, 0.08 eq.) and Pd ₂ (dba) ₃ (6 mg, 0.006 mmol, 0.04 eq.). The temperature was raised to 120°C, and the reaction in the resulting system was left to proceed for 8 hours in a nitrogen atmosphere. The completion of the reaction was determined by the LC-MS method. The reaction solution was poured into water (10 ml) and extracted with EA. The organic phase was dried and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=10:1) to give the product (14 mg, yield: 23%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.65 (s, 1H), 8.28-8.32 (d, 1H), 6.57-9.69 ( d, 1H), 3.80 (s, 3H), 3.51 (s, 3H), 2.96-3.02 (t, 3H), 2.85 (s, 4H), 1.80-2 .03 (d, 10H), 1.24-1.47 (t, 4H).

Molecular formula: C ₂₂ H ₃₀ N ₆ O, Molecular weight: 394.52, LC-MS (Pos, m/z)=395.24 [M+H] ⁺ .

Example 44

Synthesis of 6-(hydroxymethyl)-2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 62)

Sodium periodate (634.3 mg, 2.966 mmol, 1.0 eq.) was dissolved in water (10 ml), then tetrahydrofuran (10 ml) was added, cooled to 0°C in an ice bath, after which 6-(1 ,2-dihydroxyethyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (504.7 mg , 1.483 mmol, 0.5 eq.) and stirred for a reaction for 3 hours at room temperature. The completion of the reaction was determined by TLC. The mixture was extracted with dichloromethane (10 ml×3). The organic phases were combined, washed with brine (10 ml×2), dried over anhydrous sodium sulfate and filtered with suction. The filtrate was concentrated under reduced pressure to give a yellow solid (350.0 mg, yield: 38.4%).

Step 2: Synthesis of 6-(hydroxymethyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (100.0 mg, 0.324 mmol, 1.0 eq.) was dissolved in methanol (15 ml), cooled to 0°C in an ice bath, after which sodium triacetylborohydride (206.2 mg, 0.973 mmol, 3.0 eq.) was added and stirred overnight at room temperature. The completion of the reaction was determined by TLC. The reaction solution was quenched by adding H ₂ O (10 ml), stirred for 30 minutes and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=50:1-20:1) to give a yellow solid (96.9 mg, yield: 96.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.97 (s, 1H), 8.59 (s, 1H), 7.72 (s, 1H), 5.58 -5.55 (t, 1H), 4.62-4.60 (d, 2H), 3.66-3.63 (t, 4H), 1.63 (m, 4H), 0.45 (m , 4H).

Molecular formula: C ₁₇ H ₁₈ N ₄ O ₂ , Molecular weight: 310.36, LC-MS (Pos, m/z)=311.08 [M+H] ⁺ .

Example 45

Synthesis of 6-methyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 63)

6-chloro-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0.16 mmol, 1.0 eq.) and trimethylcyclotriborane (160 mg, 0.64 mmol, 4.0 eq., 50%) were dissolved in 1,4-dioxane (3 ml), after which cesium carbonate (155 mg, 0 .48 mmol, 3.0 eq.), potassium phosphate (34 mg, 0.16 mmol, 1.0 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (12 mg, 0.016 mmol, 0.1 equiv.). The temperature was raised to 120°C, and the reaction in the resulting system was left to proceed for 3 hours in a nitrogen atmosphere. The completion of the reaction was determined by the LC-MS method. The reaction solution was filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=20:1) to give a light yellow solid (10 mg, yield: 21%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.92 (s, 1H), 8.55 (s, 1H), 7.45 (s, 1H), 3.63 (m, 4H), 2.53 (s, 3H), 1.63 (m, 4H), 0.44 (m, 4H).

Molecular formula: C ₁₇ H ₁₈ N ₄ O, Molecular weight: 294.36, LC-MS (Pos, m/z)=295.09 [M+H] ⁺ .

Example 46

Synthesis of 6-methoxy-2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 64)

4-Chloro-6-methoxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (139.4 mg of crude product) was dissolved in DMF (2 ml), after which 6-aza hydrochloride was added -spiro[2,5]octane (122.5 mg, 0.83 mmol) and DIPEA (231.1 mg, 1.78 mmol). The temperature was raised to 80°C, and the reaction in the resulting system was left to proceed for 2 hours. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure, after which ethyl acetate was added, washed with water, washed with saturated brine, and dried with anhydrous sodium sulfate. The crude product was purified by preparative thin layer chromatography (DCM:MeOH=20:1) to give the product (2.28 mg, yield: 1.23%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.82 (s, 1H), 8.32 (s, 1H), 6.98 (s, 1H), 3.88 (s, 3H), 3.63-3.61 (t, 4H), 1.60 (m, 4H), 0.43 (m, 4H).

Molecular formula: C ₁₇ H ₁₈ N ₄ O ₂ , Molecular weight: 310.36, LC-MS (Pos, m/z)=311.1 [M+H] ⁺ .

Example 47

Synthesis of 6-methyl-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 65)

Material 6-chloro-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50 mg, 0. 15 mmol, 1.0 eq.) and trimethylcyclotriborane (153 mg, 0.61 mmol, 4.0 eq., 50%) were dissolved in 1,4-dioxane (3 ml), after which cesium carbonate (148 mg, 0.47 mmol, 3.0 eq.), potassium phosphate (32 mg, 0.15 mmol, 1.0 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (11 mg, 0.015 mmol , 0.1 equiv.). The temperature was raised to 120°C, and the reaction in the resulting system was left to proceed for 3 hours in a nitrogen atmosphere. The completion of the reaction was determined by the LC-MS method. The reaction solution was filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=20:1) to give a light yellow solid (15 mg, yield: 32%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.88 (s, 1H), 8.55 (s, 1H), 7.41 (s, 1H), 3.52 (s, 4H), 2.52 (s, 3H), 1.81-1.86 (m, 10H).

Molecular formula: C ₁₈ H ₂₀ N ₄ O, Molecular weight: 308.39, LC-MS (Pos, m/z)=309.07 [M+H] ⁺ .

Example 48

Synthesis of 6-methoxy-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 66)

Step 1: Synthesis of 6-methoxy-2H-pyrido[3,4-d][1,3]oxazine-2,4(1H)-dione

5-Amino-2-methoxyisonicotinic acid (500.0 mg, 2.97 mmol) was dissolved in DMF (3.5 ml). The temperature was lowered to 0° C. with an ice bath and CDI (819.7 mg, 5.05 mmol) was added. The temperature was slowly raised to room temperature and the system was stirred overnight. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure to give a product (700.0 mg of crude product), which was directly used in the next step without purification.

Step 2: Synthesis of 4-hydroxy-6-methoxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-methoxy-2H-pyrido[3,4-d][1,3]oxazine-2,4(1H)-dione (576.3 mg of crude product) was placed in a flask (50 ml) followed by the addition of ethyl cyanoacetate ( 353.0 mg, 3.12 mmol) and triethylamine (601.1 mg, 5.94 mmol) were heated to 150° C. and allowed to react overnight. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure, after which water (10 ml) was added, adjusted to pH 1 with hydrochloric acid, resulting in precipitation of a solid, and filtered with suction to obtain a product (500.0 mg of a crude product).

Step 3: Synthesis of 2,4-dichloro-6-methoxy-1,7-naphthyridine-3-carbonitrile

4-hydroxy-6-methoxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (420.0 mg crude product) was dissolved in phosphorus oxychloride (1.34 g, 8.73 mmol) , after which was added phosphopentate (phosphopentate) (807.9 mg, 3.88 mmol), heated to 100°C and left to react overnight. The completion of the reaction was determined by the LC-MS method. The reaction solution was poured into ice water (15 ml), adjusted to pH 1 with saturated sodium bicarbonate aqueous solution, and suction filtered to give the product (260.0 mg crude product).

Step 4: Synthesis of 4-chloro-6-methoxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

2,4-dichloro-6-methoxy-1,7-naphthyridine-3-carbonitrile (100.0 mg of crude product) was dissolved in TFA (2 ml) and water (1 ml), heated to 100°C and left to react in within 2 hours. Since the reaction was not completed according to TLC, TFA (2 ml) was added again, and the reaction in the resulting system was left to proceed for 2 hours at 100°C. After completion of the reaction, which was determined by TLC, the reaction solution was concentrated under reduced pressure to obtain a product (50.0 mg of crude product), which was directly used in the next step without purification.

Step 5: Synthesis of 6-methoxy-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4-Chloro-6-methoxy-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50.0 mg crude product) was dissolved in DMF (2 ml), after which 7-aza hydrochloride was added α-spiro[3,5]nonane (48.2 mg, 0.298 mmol) and DIPEA (83.0 mg, 0.639 mmol) were heated to 80° C. and allowed to react overnight. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=80:1-40:1) to give the product (4.1 mg, yield after five steps: 0.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.76 (s, 1H), 8.31 (s, 1H), 6.95 (d, 1H), 3.88 (s, 3H), 3.51-3.50 (t, 4H), 1.93-1.80 (m, 10H).

Molecular formula: C ₁₈ H ₂₀ N ₄ O ₂ , Molecular weight: 324.38, LC-MS (Pos, m/z)=325.1 [M+H] ⁺ .

Example 49

Synthesis of 6-ethyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 76)

2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (50.0 mg, 0.163 mmol) was dissolved in methanol (10 ml), after which carbon-supported palladium (10.0 mg) was added and stirred for 30 minutes at room temperature under a hydrogen atmosphere. The completion of the reaction was determined by TLC. The reaction solution was suction filtered. The filter cake was washed with a small amount of methanol. The filtrate was concentrated under reduced pressure to give the product (48.9 mg, yield: 97.2%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.94 (s, 1H), 8.60 (s, 1H), 7.44 (s, 1H), 3.65 (t, 4H), 2.85-2.79 (m, 2H), 1.63 (s, 4H), 1.23-1.27 (t, 3H), 0.44 (m, 4H).

Molecular formula: C ₁₈ H ₂₀ N ₄ O, Molecular weight: 308.39, LC-MS (Pos, m/z)=309.08 [M+H] ⁺ .

Example 50

Synthesis of 2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3,6-dicarbonitrile (Compound 77)

Step 1: Synthesis of 6-((hydroxyimino)methyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3 -carbonitrile

6-formyl-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (200 mg, 0.62 mmol, 1.0 eq.), hydroxylamine hydrochloride (66 mg, 0.93 mmol, 1.5 eq.) and potassium carbonate (112 mg, 0.8 mmol, 1.3 eq.) were dissolved in methanol (4, 5 ml) and water (1.5 ml), stirred for 2 hours at 65°C, cooled and filtered. The filter cake was washed with water (2 ml×2) to give a white solid (170 mg, yield: 81.36%).

Step 2: Synthesis of 2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3,6-dinitrile

6-((hydroxyimino)methyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (60 mg, 0.18 mmol, 1.0 eq.) was dissolved in acetic anhydride (3 ml) and stirred for 2 hours at 120°C. The reaction solution was cooled and filtered to give a yellow solid (18 mg, yield: 31.3%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.25 (s, 1H), 8.70 (s, 1H), 8.08 (s, 1H), 3.55 (m, 4H), 1.82-1.91 (m, 10H).

Molecular formula: C ₁₈ H ₁₇ N ₅ O, Molecular weight: 319.14, LC-MS (Neg, m/z) = 318.10 [M-H] ^- .

Example 51

Synthesis of 6-(aminomethyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile trifluoroacetate (Compound 78 )

6-formyl-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (100 mg, 0.31 mmol, 1.0 eq.) and tetraisopropyl titanate (440.2 mg, 1.55 mmol, 5.0 eq.) were dissolved in methanolic ammonia solution (10 ml), stirred for 18 hours at room temperature, cooled to 0° C in an ice bath, then sodium borohydride (350 mg, 1.24 mmol, 4.0 eq) was added slowly and stirred for 1 hour at room temperature. The completion of the reaction was determined by TLC. Water (1 ml) was added to stop the reaction. The crude product was purified by reverse phase column chromatography (acetonitrile: water: trifluoroacetic acid=30:100:0.05%) to give a yellow solid (16.27 mg, yield: 16.2%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.09 (s, 1H), 8.67 (s, 1H), 8.29 (br. s, 1H), 7 .66 (s, 1H), 4.25 (m, 2H), 3.63 (m, 4H), 1.88-1.83 (s, 10H).

Molecular formula: C ₁₈ H ₂₁ N ₅₀ , Molecular weight: 323.17, LC-MS (Neg, m/z) = 324.14 [M-H] ^- .

Example 52

Synthesis of 6-(methylthio)-2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 79)

Step 1: Synthesis of 4-chloro-6-(methylthio)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

2,4-dichloro-6-(methylthio)-1,7-naphthyridine-3-carbonitrile (180.0 mg, 0.666 mmol) was dissolved in a mixture of trifluoroacetate (4.0 ml) and water (1.0 ml) and stirred for 1 hour at 100°C. The completion of the reaction was determined by TLC. The mixture was concentrated under reduced pressure to give a dark red solid (180 mg crude) which was used directly in the next step.

Step 2: Synthesis of 6-(methylthio)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

4-chloro-6-(methylthio)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (167.7 mg, 0.666 mmol, 1.0 eq.), 6-aza-hydrochloride spiro[2.5]octane (118.1 mg, 0.800 mmol, 1.2 eq.) and DIPEA (688.9 mg, 5.331 mmol, 8.0 eq.) were dissolved in DMF (5 ml) and stirred for 3 hours at 80°C. The completion of the reaction was determined by TLC. The mixture was concentrated under reduced pressure to remove most of the DMF, dissolved in dichloromethane (5 ml) and concentrated under reduced pressure. This process was repeated 3 times. The crude product was purified by silica gel column chromatography (DCM:MeOH=100:1-60:1) to give a yellow solid (21.0 mg, yield: 9.66%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.95 (s, 1H), 8.56 (s, 1H), 7.37 (s, 1H), 3.65 -3.63 (t, 4H), 2.56 (s, 4H), 1.62 (m, 4H), 0.44 (m, 3H).

Molecular formula: C ₁₇ H ₁₈ N ₄ OS, Molecular weight: 326.42, LC-MS (Pos, m/z) = 327.20 [M+H] ⁺ .

Example 53

Synthesis of 2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (Compound 80)

Step 1: Synthesis of 2H-pyrido[3,2-d][1,3]oxazine-2,4(1H)-dione

3-aminopicolinic acid (460 mg, 3.3 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (8 ml), then N,N-carbonyldiimidazole (920 mg, 5.7 mmol, 1 ,7 eq.), stirred at room temperature for 24 hours and filtered. The filtrate was concentrated to give a yellow solid (235 mg, crude).

Step 2: Synthesis of 4-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

Ethyl cyanoacetate (172 mg, 1.5 mmol, 1.05 eq.) was dissolved in tetrahydrofuran (4 ml), then sodium hydride (128 mg, 3.2 mmol, 2.2 eq.) was added in portions while cooling with an ice bath and boiled at reflux for 30 minutes. During the boil, 2H-pyrido[3,2-d][1,3]oxazine-2,4-(1H)-dione (235 mg, 1.43 mmol, 1.0 eq.) was added and the reaction continued for within 18 hours. The reaction solution was cooled, poured into an ice bath, and adjusted to pH 3 to 4, whereby a solid precipitated, and filtered to give a black solid (130 mg, yield: 48.5%).

Step 3: Synthesis of 4-chloro-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

4-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (130 mg, 0.69 mmol, 1.0 eq.) was dissolved in phosphorus oxychloride (3 ml) and stirred at 80 °C for 2 hours. The reaction solution was cooled, poured into an ice bath, resulting in a black solid, and filtered to give the product as a black solid (60 mg, crude).

Step 4: Synthesis of 2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

4-chloro-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (38 mg, 0.18 mmol, 1.0 eq) and 6-aza-spiro[2.5] hydrochloride octane (30 mg, 0.20 mmol, 1.1 eq.) was dissolved in DMF (2 ml), after which triethylamine (73 mg, 0.72 mmol, 4.0 eq.) was added. The reaction solution was heated to 80° C. and stirred for 2 hours. The reaction solution was cooled and purified by reverse phase column chromatography (acetonitrile: water: ammonia solution=30:100:0.05%) to give a white solid (15.1 mg, yield: 30.0%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.66 (s, 1H), 8.49 (d, J=4 Hz, 1H), 7.67-7.64 (m, 1H), 7.60-7.58 (m, 1H), 3.88 (m, 4H), 1.60 (m, 4H), 0.41 (m, 4H).

Molecular formula: C ₁₆ H ₁₆ N ₄₀ , Molecular weight: 280.33, LC-MS (Pos, m/z) = 281.35 [M+H] ⁺ .

Example 54

Synthesis of 4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (compound 81)

The crude material, 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (300 mg, 1.46 mmol, 1.0 eq.), was dissolved in DMF (3 ml) and DIPEA (1.13 g, 8.76 mmol, 6.0 eq.) and 4-methoxy-4-methylpiperidine trifluoroacetate (496 mg, 2.04 mmol, 1.4 eq.) were added. The reaction was carried out at 80°C for 2 hours. The completion of the reaction was determined by LC-MS. The reaction solution was extracted by adding water (10 ml) and dichloromethane (10 ml×3). The organic phase was washed with water (10 ml×3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=20:1) to give a yellow solid (210 mg, yield: 48%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.99 (s, 1H), 8.65 (s, 1H), 8.33-8.34 (d, 1H) , 7.61-7.62 (d, 1H), 3.59-3.62 (m, 4H), 3.19 (s, 3H), 1.90-1.92 (d, 2H), 1 .77-1.82 (m, 2H), 1.21 (s, 3H).

Molecular formula: C ₁₆ H ₁₈ N ₄ O ₂ , Molecular weight: 298.35, LC-MS (Pos, m/z) = 299.14 [M+H] ⁺ .

Example 55

Synthesis of 6-(2-hydroxypropan-2-yl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridin-3 -carbonitrile (compound 82)

Step 1: Synthesis of 6-acetyl-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-(1-hydroxyethyl)-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (220 mg , 0.65 mmol, 1.0 eq.) was dissolved in dichloromethane (4 ml), after which Dess-Martin oxidant (552 mg, 1.3 mmol, 2.0 eq.) was added and stirred at room temperature for 18 hours. Water (10 ml) and dichloromethane (30 ml) were added. The organic phase was separated, washed with brine, dried and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane:methanol=30:1) to give the product (170 mg, yield: 77.8%).

Step 2: Synthesis of 6-acetyl-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-acetyl-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (60 mg, 0.18 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (2 ml). Methylmagnesium chloride (3.0 mol/l solution in tetrahydrofuran, 0.12 ml, 0.37 mmol, 2.0 eq.) was added under nitrogen protection at -10° C. and the mixture was heated to 0° C. and stirred for 2 hours, quenched with saturated aqueous ammonium chloride solution (5 ml) while cooling with an ice bath, after which ethyl acetate (20 ml) was added. After separation of the liquids, the organic phase was dried and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane:methanol=60:1) to give the product (17.4 mg, yield: 27.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 8.74 (s, 1H), 7.73 (s, 1H), 3.65-3.84 (m, 4H) , 1.84-2.07 (m, 10H), 1.47-1.77 (m, 6H).

Molecular formula: C ₂₀ H ₂₄ N ₄ O ₂ , Molecular weight: 352.19, LC-MS (Neg, m/z) = 351.15 [M-H] ^- .

Example 56

Synthesis of 3-cyano-2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (compound 83)

6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (92.8 mg, 0.301 mmol, 1.0 eq.) was dissolved in formic acid (10 ml) and cooled to 0°C using an ice bath. Hydrogen peroxide (30%, 2.4 ml, 24.08 mmol, 80.0 eq.) was then added dropwise. After the completion of the dropwise addition, the reaction mixture was stirred overnight with ice bath cooling. The completion of the reaction was determined by TLC. The reaction solution was filtered, the filter cake was washed with water (3 ml×3) and dried at 40° C. for 2 hours to obtain a yellow solid (46.6 mg, yield: 47.7%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 13.24 (s, 1H), 12.32 (s, 1H), 8.70 (s, 1H), 8.34 (s, 1H), 3.68 (t, 4H), 1.63 (s, 4H), 0.46 (s, 4H).

Molecular formula: C ₁₇ H ₁₆ N ₄ O ₃ , Molecular weight: 324.34, LC-MS (Pos, m/z) = 325.10 [M+H] ⁺ .

Example 57

Synthesis of 6-(2-hydroxypropan-2-yl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridin-3 -carbonitrile (compound 84)

6-acetyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (277.6 mg, 0 .86 mmol, 1.0 eq.) was dissolved in anhydrous tetrahydrofuran (20 ml) and cooled to -10° C. under nitrogen. Then a solution of methylmagnesium chloride in tetrahydrofuran (3 mol/l, 1.2 ml, 3.44 mmol, 4.0 eq.) was added dropwise. After completion of the dropwise addition, the mixture was stirred to react for 4 to 5 hours under cooling with an ice bath (0°C). The completion of the reaction was determined by TLC. The reaction was stopped by adding a saturated aqueous solution of NH ₄ Cl (10 ml). The mixture was extracted with dichloromethane (10 ml x 3). The organic phase was separated from the reaction mixture, washed with brine (10 ml × 2), dried over anhydrous sodium sulfate, filtered with suction and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=40:1-15:1) to give a yellow solid (257.4 mg, yield: 88.3%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.93 (s, 1H), 8.60 (s, 1H), 7.95 (s, 1H), 5.35 (s, 1H), 3.66-3.64 (t, 4H), 1.63 (s, 4H), 1.45 (s, 6H), 0.45 (s, 4H).

Molecular formula: C ₁₉ H ₂₂ N ₄ O ₂ , Molecular weight: 338.41, LC-MS (Pos, m/z) = 339.20 [M+H] ⁺ .

Example 58

Synthesis of 6-(hydroxymethyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (compound 85)

Step 1: Synthesis of 6-formyl-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Sodium periodate (1.63 g, 7.6 mmol, 1.0 eq.) was dissolved in water (50 ml), after which tetrahydrofuran (50 ml) was added, cooled to 0°C with an ice bath, then 6-( 1,2-dihydroxyethyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (2.7 g, 7.6 mmol, 0.5 eq.) and stirred at room temperature for 3 hours. The completion of the reaction was determined by TLC. The mixture was extracted with dichloromethane (100 ml x 3). The organic phase was washed with brine (40 ml×2), dried with anhydrous sodium sulfate, suction filtered, concentrated under reduced pressure to give the product (857.0 mg, yield: 34.89%).

Step 2: Synthesis of 6-(hydroxymethyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

6-formyl-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (100.0 mg, 0.324 mmol, 1.0 eq.) was dissolved in methanol (15 ml), cooled to 0°C using an ice bath, after which sodium borohydride (36.8 mg, 0.973 mmol, 3.0 eq.) was added and stirred at room temperature. temperature during the night. The completion of the reaction was determined by TLC, and the reaction was stopped by adding H ₂ O (10 ml). The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH=50:1-20:1) to obtain a product (96.9 mg, yield: 96.3%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.97 (s, 1H), 8.58 (s, 1H), 7.68 (s, 1H), 5.58 -5.55 (m, 1H), 4.60 (d, 2H), 3.66-3.63 (t, 4H), 1.94-1.81 (s, 10H).

Molecular formula: C ₁₈ H ₂₀ N ₄ O ₂ , Molecular weight: 324.16, LC-MS (Neg, m/z) = 323.15 [M-H] ^- .

Example 59

Synthesis of 6-(1-hydroxy-2-methylpropan-2-yl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7 -naphthyridine-3-carbonitrile (compound 86)

Step 1: Synthesis of 1-tert-butyl-3-ethyl-2-(4-(methoxycarbonyl)-5-nitropyridin-2-yl)malonate

The starting material, tert-butyl ethyl malonate (41.7 g, 222 mmol, 1.2 eq.), was dissolved in anhydrous DMF (100 ml), cooled to 0° C. with an ice bath, stirred for half an hour, after which NaH (14.8 g, 370 mmol, 2 eq.) was added, stirred for half an hour, then methyl 2-chloro-5-nitroisonicotinate (40.0 g, 185 mmol, 1.0 eq.) was slowly added and stirred at room temperature for 5 hours. The completion of the reaction was determined by TLC, and the reaction was stopped by adding H ₂ O at 0°C. The reaction solution was concentrated and extracted with EA (3×500 ml). The organic phase was dried with anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1) to give the product (35 g, yield: 51.4%).

Step 2: Synthesis of methyl 2-(2-ethoxy-2-oxoethyl)-5-nitroisonicotinate

Intermediate 1-tert-butyl-3-ethyl-2-(4-(methoxycarbonyl)-5-nitropyridin-2-yl)malonate (36.0 g, 97.8 mmol, 1.0 eq) was dissolved in trifluoroacetic acid (55.7 g, 0.5 mol, 5 eq.) at 0°C, stirred at room temperature for half an hour, and the completion of the reaction was determined by TLC. Saturated aqueous sodium bicarbonate solution was added dropwise at 0°C, stirred for half an hour and extracted with EA (3×500 ml). The organic phase was dried with anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 10:1) to give the product (23 g, yield: 87.7%).

Step 3: Synthesis of methyl 2-(1-ethoxy-2-methyl-1-oxopropan-2-yl)-5-nitroisonicotinate

The intermediate methyl 2-(2-ethoxy-2-oxoethyl)-5-nitroisonicotinate (10.7 g, 40.2 mmol, 1.0 eq.) was dissolved in DMF (100 ml), stirred at 0°C for half an hour, after which NaH (1.1 g, 44.2 mmol, 1.1 eq.) was added, slowly warmed to room temperature, stirred for half an hour, then cooled to 0 ° C, then CH ₃ I was slowly added dropwise (6.2 g, 44.2 mmol, 1.1 eq.), warmed to room temperature and stirred for 1 hour. The completion of the reaction of the starting materials was monitored by TLC, and the temperature was lowered to 0°C. The operation described above was repeated. The completion of the reaction was determined by LC-MS. The reaction was stopped by adding H ₂ O at 0°C. The reaction solution was stirred for half an hour, concentrated and extracted with EA (3×100 ml). The organic phase was dried with anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=10:1) to give the product (5.3 g, yield: 44.5%).

Step 4: Synthesis of methyl 5-amino-2-(1-ethoxy-2-methyl-1-oxopropan-2-yl)isonicotinate

The intermediate methyl 2-(1-ethoxy-2-methyl-1-oxopropan-2-yl)-5-nitroisonicotinate (5.0 g, 16.9 mmol, 1.0 eq) was dissolved in methanol (30 ml) and carbon-supported palladium (500.0 mg) was added thereto. The atmosphere was expelled from the system with hydrogen, and the reaction was carried out at room temperature. The completion of the reaction was determined by LC-MS. The reaction solution was filtered through Celite. The filtrate was concentrated, after which ethyl acetate (30 ml) was added, dried with anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure to give the product (4.3 g, yield: 95.6%).

Step 5: Synthesis of methyl 5-(2-cyanoacetamido)-2-(1-ethoxy-2-methyl-1-oxopropan-2-yl)isonicotinate

Intermediate methyl 5-amino-2-(1-ethoxy-2-methyl-1-oxopropan-2-yl)isonicotinate (4.0 g, 15.0 mmol, 1.0 eq) and cyanoacetic acid (2, 6 g, 30.0 mmol, 2.0 eq.) was dissolved in DMF (20 ml), after which EDCI (8.6 g, 45.0 mmol, 3.0 eq.) was added at 0°C and left to react at room temperature. The completion of the reaction was determined by LC-MS. The reaction solution was concentrated, then water was added and extracted with EA (3×100 ml). The organic phase was dried with anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=2:1) to give the product (3.4 g, yield: 68.1%).

Step 6: Synthesis of ethyl 2-(3-cyano-4-hydroxy-2-oxo-1,2-dihydro-1,7-naphthyridin-6-yl)-2-methylpropionate

Intermediate methyl 5-(2-cyanoacetamido)-2-(1-ethoxy-2-methyl-1-oxopropan-2-yl)isonicotinate (4.0 g, 12.0 mmol, 1.0 eq) and ethylate sodium (1.8 g, 26.4 mmol, 2.2 eq.) was successively added to ethanol (30 ml) at 0° C. and stirred at room temperature. The completion of the reaction was determined by LC-MS. The reaction solution obtained as above was added dropwise to ice water (150 ml), adjusted to pH 2, resulting in precipitation of a large amount of solid, then filtered, and the filter cake was dried to give the product (3.4 g, yield: 94.1%).

Step 7: Synthesis of ethyl 2-(4-chloro-3-cyano-2-oxo-1,2-dihydro-1,7-naphthyridin-6-yl)-2-methylpropionate

Intermediate ethyl 2-(3-cyano-4-hydroxy-2-oxo-1,2-dihydro-1,7-naphthyridin-6-yl)-2-methylpropionate (2.0 g, 6.6 mmol, 1 0 eq.) was dissolved in phosphorus oxychloride (5 ml) at 0°C, and the reaction was carried out at 100°C for half an hour. The completion of the reaction was determined by the LC-MS method, and the temperature was lowered to 0°C. The reaction solution was quenched by slowly adding water and extracted with EA (3×80 ml). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. Trifluoroacetic acid (3 ml) was added to the crude product and refluxed. The completion of the reaction of the dichloro-substituted product was monitored by LC-MS. The temperature was lowered to 0°C. Water was slowly added dropwise to the reaction solution. The mixture was extracted with EA (3×20 ml). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give the product (1.3 g, yield: 61.7%).

Step 8: Synthesis of ethyl 2-(3-cyano-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-6 -yl)-2-methylpropionate

Intermediate ethyl 2-(4-chloro-3-cyano-2-oxo-1,2-dihydro-1,7-naphthyridin-6-yl)-2-methylpropionate (800.0 mg, 2.5 mmol, 1 0 eq.) was dissolved in DMF (5 ml), then 6-aza-spiro[2.5]octane hydrochloride (404.2 mg, 2.75 mmol, 1.1 eq.) and DIPEA (1. 9 g, 15.0 mmol, 6.0 eq.) and left to react at 80°C for half an hour. The completion of the reaction was determined by TLC. The reaction solution was concentrated, after which water was added and extracted with EA (3×60 ml). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (PE:EA=3:2) to give the product (500.0 mg, yield: 50.7%).

Step 9: Synthesis of 6-(1-hydroxy-2-methylpropan-2-yl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro- 1,7-naphthyridine-3-carbonitrile

Intermediate ethyl 2-(3-cyano-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridin-6-yl) -2-methylpropionate (400.0 mg, 1.01 mmol, 1.0 eq.) was dissolved in anhydrous 2-methyltetrahydrofuran (5 ml), DIBAL-H (1.5 mol/l solution in toluene, 3.4 ml, 5.05 mmol, 5.0 eq.), then gradually warmed to room temperature, stirred for 6 hours, and quenched by adding water dropwise at 0°C, then concentrated and extracted with EA (3×60 ml). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (PE:EA=3:2) to give a yellow solid (150.0 mg, yield: 42.2%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.93 (s, 1H), 8.64 (s, 1H), 7.58 (s, 1H), 4.65 (t, 1H), 3.63-3.66 (m, 4H), 3.53 (d, 2H), 1.63 (s, 4H), 1.28 (s, 6H), 0.45 ( s, 4H).

Molecular formula: C ₂₀ H ₂₄ N ₄ O ₂ , Molecular weight: 352.44, LC-MS (Pos, m/z) = 353 [M+H] ⁺ .

Example 60

Synthesis of 6-(1-hydroxyethyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 87)

Step 1: Synthesis of 6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Sodium periodate (4.00 g, 18.70 mmol, 2.0 eq.) was dissolved in water (10 ml), after which tetrahydrofuran (100 ml) was added, cooled to 0° C. in an ice bath, then 6- (1,2-dihydroxyethyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (3, 18 g, 9.35 mmol, 1.0 eq) and stirred at room temperature for 3 hours. The completion of the reaction was determined by TLC, and the reaction solution was extracted with dichloromethane (20 ml×3). The organic phase was washed with brine (20 ml×2), dried with anhydrous sodium sulfate and filtered with suction, then the filtrate was concentrated under reduced pressure to give a yellow solid (1.50 g, yield: 52.0%).

Step 2: Synthesis of 6-(1-hydroxyethyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridin-3- carbonitrile

6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (1.00 g, 3 .24 mmol, 1.0 eq.) was dissolved in anhydrous tetrahydrofuran (50 ml), cooled to -10° C. under nitrogen, after which a solution of methylmagnesium chloride in tetrahydrofuran (3 mol/l, 4.3 ml, 12 .97 mmol, 4.0 eq.) and then stirred in an ice bath to allow the reaction to proceed for a period of 4 to 5 hours. The completion of the reaction was determined by TLC. The reaction solution was quenched by adding saturated aqueous NH ₄ Cl (20 ml) and extracted with dichloromethane (20 ml×3). The organic phase was washed with brine (20 ml×2), dried over anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=40:1-15:1) to give a yellow solid (0.95 g, yield: 90.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.96 (s, 1H), 8.59 (s, 1H), 7.78 (s, 1H), 5.50 -5.49 (d, 1H), 4.81-4.77 (m, 1H), 3.65 (s, 4H), 1.63 (s, 4H), 1.40-1.38 (d , 3H), 0.45 (s, 4H).

Molecular formula: C ₁₈ H ₂₀ N ₄ O ₂ , Molecular weight: 324.38, LC-MS (Pos, m/z) = 325.10 [M+H] ⁺ .

Example 61

Synthesis of 6-(cyclopropyl(hydroxy)methyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 90)

Intermediate 6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (200.0 mg, 0.65 mmol, 1.0 eq.) was dissolved in dry THF (5 ml), and to the reaction solution thus obtained at −10° C., under nitrogen atmosphere, was added cyclopropylmagnesium bromide (1.95 ml, 1.95 mmol, 3 0 equiv.). The reaction mixture was slowly warmed to 0°C and stirred for 1 hour. After completion of the reaction, the reaction solution was quenched by adding a saturated aqueous ammonium chloride solution, concentrated and extracted with ethyl acetate (3×30 ml). The organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (DCE (dichloroethane): MeOH=100:1-75:1) to give an off-white solid (92 mg, yield: 40.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.96 (br, 1H), 8.60 (s, 1H), 7.74 (s, 1H), 5.44 (s, 1H), 4.24-4.27 (m, 1H), 3.65 (t, 4H), 1.63 (s, 4H), 1.12-1.18 (m, 1H), 0.45 (s, 4H), 0.40-0.43 (m, 4H).

Molecular formula: C ₂₀ H ₂₂ N ₄ O ₂ , Molecular weight: 350.42, LC-MS (m/z) = 351 [M+H] ⁺ .

Example 62

Synthesis of 4-methoxy-4-methylpiperidine trifluoroacetate intermediate

Step 1: Synthesis of tert-butyl 4-hydroxy-4-methylpiperidine-1-carboxylate

The crude material tert-butyl-4-oxo-piperidine-1-carboxylate (5.0 g, 25 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (25 ml), after which methylmagnesium chloride ( 9 ml, 27 mmol, 1.1 eq.). The reaction was carried out for 2 hours, and the completion of the reaction was determined by TLC. The pH of the reaction solution was adjusted to 4 by adding dilute hydrochloric acid, after which water (30 ml) was added and extracted with ethyl acetate (30 ml×3). The organic phase was dried, filtered and concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE:EA=5:1) to give the product (5.2 g, yield: 96%).

Step 2: Synthesis of tert-butyl 4-methoxy-4-methylpiperidine-1-carboxylate

The intermediate tert-butyl 4-hydroxy-4-methylpiperidine-1-carboxylate (500 mg, 2.32 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (5 mL), after which sodium hydride (186 mg, 4.0 equiv.) was added. 64 mmol, 2.0 eq.), and the reaction proceeded for 1 hour, after which was added iodomethane (659 mg, 4.64 mmol, 2.0 eq.), left to react for 8 hours. The completion of the reaction was determined by TLC. Water (10 ml) was added to the reaction flask, and the reaction solution was extracted with ethyl acetate (20 ml×3). The organic phase was dried, filtered and concentrated under reduced pressure; the crude product was purified by silica gel column chromatography (PE:EA = 20:1) to give the product (306 mg, yield: 57%).

Step 3: Synthesis of 4-methoxy-4-methylpiperidine trifluoroacetate

The intermediate tert-butyl 4-methoxy-4-methylpiperidine-1-carboxylate (500 mg, 2.18 mmol, 1.0 eq.) was dissolved in dichloromethane (4 ml), after which trifluoroacetic acid (3 ml) and left to react for 1 hour. The completion of the reaction was determined by TLC; the reaction solution was concentrated under reduced pressure to give the product (530 mg, yield: 100%).

Example 63

Synthesis of 6-(1-hydroxyprop-2-yn-1-yl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7 -naphthyridine-3-carbonitrile (Compound 91)

Intermediate 6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-acetonitrile (200.0 mg, 0.65 mmol, 1.0 eq.) was dissolved in anhydrous THF (5 ml), and ethynylmagnesium bromide (6.48 ml, 3.25 mmol, 5 0 equiv.). The reaction solution was slowly warmed to 0° C. and stirred for 1 hour. Upon completion of the reaction, the solution was quenched by adding a saturated aqueous ammonium chloride solution, concentrated and extracted with ethyl acetate (3×30 ml). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated; the crude product was purified by silica gel column chromatography (DCE:MeOH=100:1-75:1) to give a white solid (40 mg, yield: 18.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.05 (br. s, 1H), 8.61 (s, 1H), 7.86 (s, 1H), 6 .36 (d, 1H), 5.39-5.42 (m, 1H), 3.64 (s, 4H), 3.51 (d, 1H), 1.64 (s, 4H), 0. 45 (m, 4H).

Molecular formula: C ₁₉ H ₁₈ N ₄ O ₂ , Molecular weight: 334.38, LC-MS (m/z) = 335 [M+H] ⁺ .

Example 64

Synthesis of 2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-6-(2,2,2-trifluoro-1-hydroxyethyl)-1,2-dihydro-1,7 -naphthyridine-3-carbonitrile (Compound 92)

6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (300 mg, 0.973 mmol, 1.0 eq.) was dissolved in anhydrous N,N-dimethylacetamide (20 ml), cooled to -10°C under nitrogen atmosphere, after which TMSCF ₃ (1.44 ml, 9.730 mmol, 10.0 eq.) was added, anhydrous TBAF in tetrahydrofuran (0.96 ml, 0.96 mmol) was added dropwise, and then stirred at -10° C. overnight for a reaction to proceed. The completion of the reaction was determined by TLC. Then, dichloromethane (20 ml) was added to the reaction solution, washed with saturated brine (20 ml × 2) and water (20 ml × 4), dried with anhydrous magnesium sulfate, filtered with suction, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCE (dichloroethane): MeOH=30:1-15:1) to give a yellow solid (116 mg, yield: 31.5%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.11 (s, 1H), 8.65 (s, 1H), 7.87 (s, 1H), 7.13 -7.12 (d, 1H), 5.24-5.22 (t, 1H), 3.64 (s, 4H), 1.63 (s, 4H), 0.45 (m, 4H).

Molecular formula: C ₁₈ H ₁₇ F ₃ N ₄ O ₂ , Molecular weight: 378.36, LC-MS (Pos, m/z) = 379.20 [M+H] ⁺ .

Example 65

Synthesis of ethyl 1-(3-cyano-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridin-6-yl)acetate (Compound 93)

6-(1-hydroxyethyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (100 mg , 0.308 mmol, 1.0 eq.) was dissolved in dichloromethane (15 ml), cooled to 0°C in an ice bath, after which acetic anhydride (63 mg, 0.616 mmol, 2.0 eq.), triethylamine (125 mg , 1.233 mmol, 4.0 eq.) and DMAP (19 mg, 0.154 mmol, 0.5 eq.), then stirred for reaction for 5 minutes in an ice bath (0°C) and then warmed to room temperature and stirred for 2 hours. The completion of the reaction was determined by TLC. The reaction solution was quenched by adding saturated aqueous NaHCO ₃ (10 ml) and extracted with dichloromethane (10 ml×3). The organic phases were combined, washed with brine (10 ml×2), dried over anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin layer chromatography (DCM:MeOH=10:1) to give an orange-yellow solid (98 mg, yield: 86.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.05 (s, 1H), 8.64 (s, 1H), 7.62 (s, 1H), 5.90 -5.85 (q, 1H), 3.64 (s, 4H), 2.06 (s, 3H), 1.63 (s, 4H), 1.55-1.53 (d, 3H), 0.45 (s, 4H).

Molecular formula: C ₂₀ H ₂₂ N ₄ O ₃ , Molecular weight: 366.42, LC-MS (Pos, m/z) = 367.20 [M+H] ⁺ .

Example 66

Synthesis of 6-(1-hydroxyethyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 94)

6-formyl-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (530.0 mg, 1 .65 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (10 ml), cooled to -10°C, then 3 mol/l methylmagnesium chloride (1.7 ml, 5.1 mmol, 3.0 eq. ) and stirred for 2 hours in an ice bath. The completion of the reaction was determined by TLC. The reaction solution was quenched by adding a saturated ammonium chloride aqueous solution (10 ml) and extracted with ethyl acetate (20 ml×2). The organic phases were combined and concentrated under reduced pressure; the crude product was purified by silica gel column chromatography (DCM:MeOH=50:1-20:1) to give the product (56.1 mg, yield: 10.1%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.92 (s, 1H), 8.58 (s, 1H), 7.73 (s, 1H), 5.49 (d, 1H, J=4.64 Hz), 4.71-4.82 (m, 1H), 3.50-3.61 (m, 4H), 1.79-1.98 (m, 10H ), 1.34 (d, 3H, J=6.48 Hz).

Molecular formula: C ₁₉ H ₂₂ NO ₂ , Molecular weight: 338.17, LC-MS (Neg, m/z) = 337.15 [M-H] ^- .

Example 67

Synthesis of 6-acetyl-2-oxo-4-(6-aza-spiro[2,5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compounds 95)

6-(1-hydroxyethyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (0, 85 g, 2.62 mmol, 1.0 eq.) was dissolved in anhydrous dichloromethane (60 ml), cooled to 0°C under nitrogen atmosphere, after which Dess-Martin oxidizer (2.22 g, 5.24 mmol, 2.0 eq.) and stirred at room temperature overnight. The completion of the reaction was determined by TLC. The reaction solution was quenched with saturated aqueous NaHCO ₃ (20 ml) and suction filtered through celite. The filtrate was extracted with dichloromethane (20 ml x 3). After liquid separation, the organic phase was washed with brine (20 ml×2), dried over anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=80:1-50:1) to give the product as a yellow solid (0.35 g, yield: 41.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.35 (s, 1H), 8.72 (s, 1H), 8.25 (s, 1H), 3.68 (t, 4H), 2.64 (s, 3H), 1.64 (s, 4H), 0.46 (s, 4H).

Molecular formula: C ₁₈ H ₁₈ N ₄ O ₂ , Molecular weight: 322.37, LC-MS (Pos, m/z) = 323.10 [M+H] ⁺ .

Example 68

Synthesis of 3-cyano-N-methyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-6-formamide (Compound 96)

Intermediate 3-cyano-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (130 mg, 0 .4 mmol, 1.0 eq.) was dissolved in N,N-dimethylformamide (2 ml), after which N,N-diisopropylethylamine (155 mg, 1.2 mmol, 3.0 eq.) and HATU (228 mg , 0.6 mmol, 1.5 eq.) and left to react at room temperature for 1 hour, after which methylamine in methanol (0.5 ml) was added and left to react at room temperature for 2 hours. The completion of the reaction was determined by LC-MS. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH=15:1) to obtain a product (50 mg, yield: 37%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.19 (s, 1H), 8.72-8.73 (m, 1H), 8.32 (s, 1H) , 3.66-3.68 (m, 4H), 2.83-2.84 (d, 3H), 1.64 (m, 4H), 0.46 (s, 4H).

Molecular formula: C ₁₈ H ₁₉ N ₅ O ₂ , Molecular weight: 337.38 LC-MS (Pos, m/z) = 338.16 [M+H] ⁺ .

Example 69

Synthesis of 3-cyano-N-(2-hydroxyethyl)-2-oxo-4-(6-aza-spiro[2.5]oct-6-yl)-1,2-dihydro-1,7-naphthyridin-6 -carboxamide (Compound 97)

Intermediate 3-cyano-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (162 mg, 0 .5 mmol, 1.0 eq.) was dissolved in N,N-dimethylformamide (2 ml), after which triethylamine (152 mg, 1.5 mmol, 3.0 eq.) and HATU (285 mg, 0.75 mmol, 1.5 eq.) and left to react at room temperature for 1 hour, after which ethanolamine (31 mg, 0.5 mmol, 1.0 eq.) was added and left to react at room temperature for 2 hours. The completion of the reaction was determined by LC-MS. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH=15:1) to obtain a product (60 mg, yield: 33%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.24 (s, 1H), 8.61-8.65 (m, 2H), 8.33 (s, 1H) , 4.77-4.80 (m, 1H), 3.66-3.69 (m, 4H), 3.51-3.56 (m, 2H), 3.38-3.42 (m, 2H), 1.65 (m, 4H), 0.46 (s, 4H).

Molecular formula: C ₁₉ H ₂₁ N ₅ O ₃ , Molecular weight: 367.41 LC-MS (Pos, m/z) = 368.28 [M+H] ⁺ .

Example 70

Synthesis of 4-(4-hydroxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 106)

Step 1: Synthesis of 4-methylpiperidin-4-ol trifluoroacetate

The crude material, tert-butyl 4-hydroxy-4-methylpiperidine-1-carboxylate (380 mg, 1.77 mmol, 1.0 eq.), was dissolved in dichloromethane (5 ml), after which trifluoroacetic acid was added at 0° C. acid (3 ml) and left to react for 1 hour. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure to give the product (404 mg, yield: 100%).

Step 2: Synthesis of 4-(4-hydroxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

The intermediate 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (200 mg, 0.97 mmol, 1.0 eq.) was dissolved in DMF (3 ml), after which N,N-diisopropylethylamine (753 mg, 5.87 mmol, 6.0 eq.) and 4-methylpiperidin-4-ol trifluoroacetate (312 mg, 1.36 mmol, 1.4 eq.) and the reaction was carried out at 80 °C for 2 hours. The completion of the reaction was determined by LC-MS. The reaction solution was added dropwise to water (5 ml), stirred for 10 minutes, and filtered with suction. The filter cake was then suspended in dichloromethane (5 ml), suction filtered and dried at 50° C. to give the product as a yellow solid (136 mg, yield: 49%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.95 (s, 1H), 8.62-8.65 (d, 1H), 8.33-8.34 ( d, 1H), 7.59-7.61 (d, 1H), 4.56-4.61 (m, 1H), 3.68-3.79 (m, 2H), 3.58-3, 62 (d, 2H), 1.77-1.83 (m, 2H), 1.66-1.76 (m, 2H), 1.24 (s, 3H).

Molecular formula: C ₁₅ H ₁₆ N ₄ O ₂ , Molecular weight: 284.32 LC-MS (Pos, m/z) = 285.13 [M+H] ⁺ .

Example 71

Synthesis of 6-ethyl-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 107)

Step 1: Synthesis of 6-chloro-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 4,6-dichloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (2.0 g, 8.33 mmol, 1.0 eq.) was dissolved in DMF (10 ml) , after which DIPEA (6.45 g, 50 mmol, 6.0 eq.) and 4-methoxy-4-methylpiperidine trifluoroacetate (2.2 g, 9.16 mmol, 1.1 eq.) were added and left to react at 80°C for 2 hours. The completion of the reaction was determined by LC-MS. Then, water (10 ml) was added to the reaction solution, which was extracted with dichloromethane (10 ml×3). The organic phase was washed with water (10 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the product as a yellow solid (2.7 g, crude).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.11 (s, 1H), 8.45 (s, 1H), 7.61 (s, 1H), 3.61 -3.59 (m, 4H), 3.18 (s, 3H), 1.91-1.88 (m, 2H), 1.81-1.76 (m, 2H), 1.21 (s , 3H).

Molecular formula: C ₁₆ H ₁₇ N ₄ O ₂ Cl, Molecular weight: 332.79 LC-MS (Pos, m/z) = 333.7 [M+H] ⁺ .

Step 2: Synthesis of 4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 6-chloro-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (2.7 g crude, 8 11 mmol, 1.0 eq.) was dissolved in 1,4-dioxane (20 ml) and H ₂ O (5 ml), after which potassium vinyltrifluoroborate (1.63 g, 12.17 mmol, 1.5 eq. ), cesium carbonate (3.965 g, 12.17 mmol, 1.5 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (297 mg, 0.41 mmol, 0.05 eq.), and left to react at 100°C for 8 hours in a nitrogen atmosphere. The completion of the reaction was determined by LC-MS. Then, water (20 ml) was added to the reaction solution, which was extracted with dichloromethane (30 ml×3). The organic phase was dried with anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH=70:1) to give the product as a yellow solid (1.15 g, yield: 43%) .

Step 3: Synthesis of 6-ethyl-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (150 mg, 0.46 mmol, 1 0 eq.) was dissolved in methanol (5 ml). Pd/C (100 mg) was added to the reaction solution. The air in the system was replaced with hydrogen three times, and the reaction was carried out for 1 hour in a hydrogen atmosphere. The completion of the reaction was determined by LC-MS. The reaction solution was filtered with suction, and the filtrate was concentrated under reduced pressure to obtain the product (120 mg, yield: 80%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.89 (s, 1H), 8.59 (s, 1H), 7.41 (s, 1H), 3.60 -3.62 (m, 4H), 3.19 (s, 3H), 2.79-2.84 (m, 2H), 1.89-1.93 (m, 2H), 1.75-1 .82 (m, 2H), 1.22-1.27 (m, 6H).

Molecular formula: C ₁₈ H ₂₂ N ₄ O ₂ , Molecular weight: 326.40 LC-MS (Pos, m/z) = 327.26 [M+H] ⁺ .

Example 72

Synthesis of 6-(1-hydroxyethyl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 108)

Step 1: Synthesis of 6-(1,2-dihydroxyethyl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (500 mg, 1.542 mmol, 1.0 eq.) was dissolved in tert-butanol (10 ml) and water (10 ml), after which methanesulfonamide (147 mg, 1.542 mmol, 1.0 eq.) and AD-mix-β (6.0 g) were added and left react at room temperature for 12 hours. The completion of the reaction was determined by LC-MS. Then, water (10 ml) was added to the reaction solution, which was extracted with dichloromethane (30 ml×3). The organic phase was dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the product (552 mg, yield: 100%).

Step 2: Synthesis of 6-formyl-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 6-(1,2-dihydroxyethyl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (552 mg, 1.542 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (10 ml) and water (2 ml), after which sodium periodate (650 mg, 3.084 mmol, 2.0 eq.) was added and left to react for 4 hours. The completion of the reaction was determined by LC-MS. Then, water (10 ml) was added to the reaction solution, which was extracted with ethyl acetate (20 ml×3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH = 60:1) to give the product as a yellow solid (160 g, yield from two synthetic steps: 32%).

Step 3: Synthesis of 6-(1-hydroxyethyl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 6-formyl-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (160 mg, 0.49 mmol, 1 0 eq.) was dissolved in tetrahydrofuran (5 ml), then methylmagnesium chloride (1 ml) was added dropwise at 0° C. and left to react for 1 hour. The completion of the reaction was determined by LC-MS. Then, water (10 ml) was added to the reaction solution, which was extracted with ethyl acetate (20 ml×3). The organic phase was dried with anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (DCM:MeOH=40:1) to give the product (108 mg, yield: 64%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.93 (s, 1H), 8.58 (s, 1H), 7.72 (s, 1H), 5.48 -5.49 (d, 1H), 4.75-4.81 (m, 1H), 3.56-3.65 (m, 4H), 3.20 (s, 3H), 1.91-1 .95 (m, 2H), 1.73-1.79 (m, 2H), 1.38-1.40 (d, 3H), 1.23 (s, 3H).

Molecular formula: C ₁₈ H ₂₂ N ₄ O ₃ , Molecular weight: 342.40 LC-MS (Pos, m/z) = 343.17 [M+H] ⁺ .

Example 73

Synthesis of 6-(2-hydroxypropan-2-yl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 109)

Step 1: Synthesis of 6-acetyl-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 6-(1-hydroxyethyl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (187 mg, 0. 55 mmol, 1.0 eq.) was dissolved in dry dichloromethane (5 ml). The temperature was lowered to 0-5° C. and Dess-Martin oxidizer (463.5 mg, 1.10 mmol, 2.0 eq.) was added. Upon completion of the addition, the temperature of the mixture was naturally allowed to rise to room temperature and allowed to react for 2 hours. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (MeOH:DCM=1:100-1:50) to give the product (185.8 mg, yield: 100%).

Step 2: Synthesis of 6-(2-hydroxypropan-2-yl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3- carbonitrile

Intermediate 6-acetyl-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (185.8 mg, 0.55 mmol , 1.0 eq.) was dissolved in N,N-dimethylacetamide (3 ml) and the temperature was lowered to -10-0°C. A solution of methylmagnesium chloride in tetrahydrofuran at a concentration of 3 mol/l (0.6 ml, 3.0 eq.) was added dropwise to the resulting mixture under nitrogen atmosphere. Upon completion of the addition, the mixture was allowed to warm to room temperature and stirred overnight. TLC analysis indicated the presence of a large amount of starting materials remaining. To the reaction solution was added a solution of methyl magnesium chloride in tetrahydrofuran at a concentration of 3 mol/l (0.6 ml, 3.0 equiv.), and the reaction was left to proceed for 3 hours, a solution of methyl magnesium chloride in tetrahydrofuran at a concentration of 3 mol/l (0.6 ml, 3.0 eq.), the reaction was left to proceed for 2 hours, cooled to 0-10°C, adjusted to pH 5-6 with acetic acid and concentrated. The crude product was purified by silica gel column chromatography (MeOH:DCM=1:100-1:70) to give the product (63.9 mg, yield: 32.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.91 (s, 1H), 8.59 (s, 1H), 7.89 (s, 1H), 5.35 (s, 1H), 3.62-3.60 (m, 4H), 3.20 (s, 3H), 1.95-1.92 (m, 2H), 1.80-1.73 (m , 2H), 1.45 (s, 6H), 1.23 (s, 3H).

Molecular formula: C ₁₉ H ₂₄ N ₄ O ₃ , Molecular weight: 356.43 LC-MS (Pos, m/z) = 357.25 [M+H] ⁺ .

Example 74

Synthesis of (S)-4-(3-(1Н-1,2,4-triazol-1-yl)pyrrolidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3- carbonitrile (Compound 112)

Step 1: Synthesis of tert-butyl-(S)-3-(1H-1,2,4-triazol-1-yl)pyrrolidine-1-carboxylate

The starting material, tert-butyl-(R)-3-hydroxypyrrolidine-1-carboxylate (1.0 g, 5.34 mmol, 1.0 eq.), was dissolved in anhydrous tetrahydrofuran (20 ml) and 1H-1, 2,4-triazole (368 mg, 5.34 mmol, 1.0 eq.) and triphenylphosphine (2.8 g, 10.68 mmol, 2.0 eq.). Upon completion of the addition, the temperature was lowered to 0° C. and diethyl azodicarboxylate (1.86 g, 10.68 mmol, 2.0 eq.) was added dropwise. Upon completion of the addition, the reaction was carried out at room temperature for 12 hours. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (EA:PE=1:15-1:2) to give the product (559 mg, yield: 44%).

Step 2: Synthesis of (S)-1-(pyrrolidin-3-yl)-1H-1,2,4-triazole hydrochloride

The intermediate tert-butyl-(S)-3-(1H-1,2,4-triazol-1-yl)pyrrolidine-1-carboxylate (559 mg, 2.34 mmol, 1.0 eq) was dissolved in anhydrous methanol (5 ml), after which a 30% solution of hydrogen chloride in ethanol (5 ml) was added and left to react at room temperature for 2 hours until a white solid precipitated. The completion of the reaction was determined by TLC. The reaction solution was concentrated under reduced pressure, after which water (10 ml) was added and extracted with EA (3×5 ml). The aqueous phase was lyophilized to give an off-white solid (410 mg, yield: 100%).

Step 3: Synthesis of (S)-4-(3-(1H-1,2,4-triazol-1-yl)pyrrolidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine -3-carbonitrile

The intermediate 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (346.2 mg, 1.68 mmol, 1.0 eq.) was dissolved in DMF (3 ml) and added (S)-1-(pyrrolidin-3-yl)-1H-1,2,4-triazole hydrochloride (410 mg, 2.34 mmol, 1.4 eq) and DIPEA (1.1 g, 8.4 mmol, 5.0 eq.). Upon completion of the addition, the temperature was raised to 80°C and the reaction was carried out for 2 hours. The completion of the reaction was determined by the LC-MS method, and the temperature was lowered to room temperature. The reaction solution was separated by reverse phase column chromatography (0.1% aqueous hydrochloric acid: acetonitrile = 70:30) to give the product (226 mg, yield: 43.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.68 (s, 1H), 8.70 (s, 1H), 8.62 (s, 1H), 8.28 -8.27 (d, 1H), 8.04 (s, 1H), 7.98-7.97 (s, 1H), 5.33-5.30 (m, 1H), 4.61-4 .57 (m, 1H), 4.35-4.28 (m, 2H), 4.23-4.17 (m, 1H), 2.55-2.51 (m, 1H), 2.50 -2.47 (m, 1H).

Molecular formula: C ₁₅ H ₁₃ N ₇ O, Molecular weight: 307.21 LC-MS (Pos, m/z) = 307.98 [M+H] ⁺ .

Example 75

Synthesis of 2-oxo-4-(3-(thiazol-2-yl)pyrrolidin-1-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 113)

Step 1: Synthesis of tert-butyl-3-(thiazol-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate

Starting material, tert-butyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (1.0 g, 3.39 mmol, 1.0 eq.) was dissolved in 1,4-dioxane (20 ml) and 2-bromothiazole (666.6 mg, 4.06 mmol, 1.2 eq.), anhydrous sodium carbonate was added (897.5 mg, 8.47 mmol, 2.5 eq.) and water (4 ml). The air in the system was purged with nitrogen three times and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (247.8 mg, 0.34 mmol, 0.1 eq.) was added to the reaction. The air in the system was expelled three times with nitrogen, the temperature was raised to 80°C and left to react for 2 hours. The completion of the reaction was determined by TLC. The temperature was lowered to room temperature. Water (10 ml) was added to the mixture and extracted with ethyl acetate (50 ml x 3). The mixture was separated, the organic phases were combined, dried over anhydrous sodium sulfate and filtered. The mother liquor was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (EA:PE=1:30-1:20) to give a pale yellow oily product (719 mg, yield: 84.2%).

Step 2: Synthesis of tert-butyl 3-(thiazol-2-yl)pyrrolidine-1-carboxylate

The intermediate tert-butyl-3-(thiazol-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (719 mg, 2.84 mmol, 1.0 eq.) was dissolved in anhydrous ethanol (20 ml ), after which 10% palladium on a carbon carrier (0.5 g) was added. Upon completion of the addition, the air in the system was expelled with hydrogen three times, the temperature was raised to 50°C, and the reaction proceeded for 12 hours. The completion of the reaction was determined by TLC, and the temperature was lowered to room temperature. The reaction solution was filtered, and the filter cake was rinsed with ethanol. The mother liquor was concentrated under reduced pressure to give a pale yellow oily product (653 mg, yield: 90.5%).

Step 3: Synthesis of 2-(pyrrolidin-3-yl)thiazole hydrochloride

The intermediate tert-butyl 3-(thiazol-2-yl)pyrrolidine-1-carboxylate (653 mg, 2.57 mmol, 1.0 eq.) was dissolved in anhydrous methanol (2 ml), after which 30% a solution of hydrogen chloride in ethanol (5 ml) and left to react at room temperature for 2 hours. The completion of the reaction was determined by LC-MS. The reaction solution was concentrated under reduced pressure to give a product (880 mg of crude product), which was directly introduced into the next step without purification.

Step 4: Synthesis of 2-oxo-4-(3-(thiazol-2-yl)pyrrolidin-1-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (210 mg, 1.02 mmol, 1.0 eq.) was dissolved in DMF (2 ml) and into the reaction mixture 2-(pyrrolidin-3-yl)thiazole hydrochloride (195 mg, 1.02 mmol, 1.0 eq) and DIPEA (792.7 mg, 6.14 mmol, 6.0 eq) were added. Upon completion of the addition, the temperature was raised to 80°C, and the mixture was left to react for 2 hours. The completion of the reaction was determined by the LC-MS method, and the temperature was lowered to room temperature. The mixture was purified by preparative HPLC (high performance liquid chromatography, also called high pressure liquid chromatography) (0.1% aqueous trifluoroacetic acid: acetonitrile = 70:30), lyophilized, dissolved by adding water (1 ml), adjusted to approximately pH 8 by adding a saturated sodium carbonate aqueous solution until a solid precipitated, and then filtered. The filter cake was dried to give the product (92 mg, yield: 27.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.63 (s, 1H), 8.61 (s, 1H), 8.28-8.27 (d, 1H) , 7.99-7.98 (d, 1H), 7.79-7.78 (d, 1H), 7.70-7.69 (d, 1H), 4.48-4.44 (m, 1H), 4.34-4.30 (m, 1H), 4.27-4.15 (m, 2H), 4.06-4.00 (m, 1H), 2.50 (m, 1H) , 2.30-2.27 (m, 1H).

Molecular formula: C ₁₆ H ₁₃ N ₅ OS, Molecular weight: 323.37 LC-MS (Pos, m/z) = 346.14 [M+H] ⁺ .

Example 76

Synthesis of 2-(methylthio)-6-oxo-8-(6-aza-spiro[2,5]octan-6-yl)-5,6-dihydropyrido[3,2-d]pyrimidine-7-carbonitrile (Compound 129)

Step 1: Synthesis of 6-(methylthio)-2H-pyrimido[5,4-d][1,3]oxazine-2,4-(1H)-dione

4-Amino-2-(methylthio)-pyrimidine-5-carboxylic acid (2.00 g, 10.80 mmol, 1.0 eq.) was dissolved in anhydrous tetrahydrofuran (100.0 ml), after which the N,N'-carbonyldiimidazole (3.50 g, 21.60 mmol, 2.0 eq.) was stirred at room temperature, reacting for 16 hours. The completion of the reaction was determined by TLC. The solid was filtered off and the filtrate was taken directly to the next reaction.

Step 2: Synthesis of 8-hydroxy-2-(methylthio)-6-oxo-5,6-dihydropyrido[3,2-d]pyrimidine-7-carbonitrile

Sodium hydride (60%, 1.88 g, 46.96 mmol, 4.35 eq.) was added slowly to anhydrous tetrahydrofuran (100.0 ml), stirred for 10 minutes and slowly added dropwise while cooling with an ice bath ethyl cyanoacetate (3.50 g, 30.95 mmol, 2.86 eq.). The mixture was stirred under nitrogen at 75° C. for 20 minutes, then 6-(methylthio)-2H-pyrimido[5,4-d][1,3]oxazine-2,4-(1H)- was slowly added dropwise. dione in THF and then stirred at 75° C. overnight under nitrogen. The completion of the reaction was determined by TLC. Then, ice water (30 ml) was added to the reaction solution, adjusted to pH 3-4 with concentrated hydrochloric acid, and filtered with suction to obtain a yellow solid (880.0 mg, yield after two synthetic steps: 34.8%).

Step 3: Synthesis of 8-chloro-2-(methylthio)-6-oxo-5,6-dihydropyrido[3,2-d]pyrimidine-7-carbonitrile

8-hydroxy-2-(methylthio)-6-oxo-5,6-dihydropyrido[3,2-d]pyrimidine-7-carbonitrile (705 mg, 3.01 mmol) was added to a mixed solvent consisting of POCl ₃ ( 10 ml) and 1,2-dichloroethane (20 ml) and stirred at 75°C; the reaction proceeded for 3-4 hours in a nitrogen atmosphere. The completion of the reaction was determined by TLC. The reaction solution was cooled in an ice bath, after which ice water (10 ml) was added, followed by stirring and filtering with suction to obtain the product (400.0 mg, yield: 52.6%).

Step 4: Synthesis of 2-(methylthio)-6-oxo-8-(6-aza-spiro[2,5]octan-6-yl)-5,6-dihydropyrido[3,2-d]pyrimidine-7- carbonitrile

8-chloro-2-(methylthio)-6-oxo-5,6-dihydropyrido[3,2-d]pyrimidine-7-carbonitrile (400 mg, 1.58 mmol, 1.0 eq.), hydrochloride 6- aza-spiro[2.5]octane (280.5 mg, 1.90 mmol, 1.2 eq.) and DIPEA (1636.7 mg, 12.66 mmol, 8.0 eq.) were added to DMF (20 ml) and stirred at 80°C, carrying out the reaction for 3 hours. Upon completion of the reaction according to TLC, the mixture was concentrated under reduced pressure, after which EA (5 ml) and water (10 ml) were added, stirred for 2 hours and filtered with suction. The filter cake was suspended by repeated addition of EA (5 ml) over 1 hour, and the product was obtained by filtration (59.0 mg, yield: 11.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.58 (s, 1H), 8.69 (s, 1H), 3.90 (t, 4H), 2.53 (s, 3H), 1.61 (t, 4H), 0.42 (s, 4H).

Molecular formula: C ₁₆ H ₁₇ N ₅ OS, Molecular weight: 327.41 LC-MS (Pos, m/z) = 328.10 [M+H] ⁺ .

Example 77

Synthesis of 6-(cyclopropyl(hydroxy)methyl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 130)

Intermediate 6-formyl-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (500 mg, 1.53 mmol, 1 0 eq.) was dissolved in anhydrous tetrahydrofuran (20 ml) and the temperature was lowered under nitrogen to -10°C. A solution of cyclopropylmagnesium bromide in tetrahydrofuran at a concentration of 1 mol/l (4.6 ml, 4.60 mmol, 3 eq.) was added dropwise to the reaction mixture. Upon completion of the addition, the reaction was carried out at 0°C for 3 hours. After that, according to LC-MS data, 20% of the starting materials remained in the mixture, so a solution of cyclopropylmagnesium bromide in tetrahydrofuran at a concentration of 1 mol/l (3 ml, 3 mmol, 2 eq.) was again added, and the reaction was continued for a period of from 2 up to 3 hours. After that, according to LC-MS, 10% of the starting materials remained in the mixture, so the pH of the mixture was adjusted to approximately 5-6 by adding acetic acid, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (MeOH:DCM=1:100-1:40) to give the product (225.7 mg, yield: 40.0%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.93 (s, 1H), 8.59 (s, 1H), 7.68 (s, 1H), 5.42 -5.40 (d, 1H), 4.24-4.22 (m, 1H), 3.63-3.60 (m, 4H), 3.19 (s, 3H), 1.95-1 .91 (m, 2H), 1.79-1.72 (m, 2H), 1.23 (s, 3H), 1.23 (s, 1H), 0.42 (m, 4H).

Molecular formula: C ₂₀ H ₂₄ N ₄ O ₃ , Molecular weight: 368.44 LC-MS (Pos, m/z) = 369.40 [M+H] ⁺ .

Example 78

Synthesis of 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-N-methyl-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxamide (Compound 131)

Step 1: Synthesis of 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid

Intermediate 6-formyl-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (681 mg, 2.09 mmol, 1 0 eq.) was dissolved in formic acid (5 ml) and the temperature was lowered to -5 to 0°C. 30% hydrogen peroxide (1.32 ml, 10.44 mmol, 5 eq.) was added to the reaction mixture. Upon completion of the addition, the reaction was carried out at 0° C. for 12 hours, then 30% hydrogen peroxide (1.32 ml, 10.44 mmol, 5 eq.) was added and left to react at room temperature for 2-3 hours. The completion of the reaction was determined by TLC. The reaction solution was poured into methyl tert-butyl ether (50 ml) resulting in the precipitation of a pale yellow solid which was filtered off. The filter cake was dried to give the product (300 mg, yield: 42.0%).

Step 2: Synthesis of 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-N-methyl-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxamide

Intermediate 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (300 mg, 0.88 mmol, 1.0 eq.) was dissolved in anhydrous N,N-dimethylacetamide (3 ml) and then DIPEA (565.8 mg, 4.38 mmol, 5.0 eq.) was added. Upon completion of the addition, the reaction mixture was cooled to 0°C, then HATU (499.7 mg, 1.31 mmol, 1.5 eq.) was added, stirred at room temperature for a time ranging from half an hour to 1 hour, after which methylamine hydrochloride (118.2 mg, 1.75 mmol, 2.0 eq.) and allowed to react at room temperature for 1 hour, resulting in the formation of a solid precipitate. The completion of the reaction was determined by TLC. Water (50 ml) was added to the reaction solution, stirred for 5 minutes, and filtered. The filter cake was rinsed with water, added to ethyl acetate (10 ml) and refluxed for 1 hour, filtered hot and dried to give the product (199 mg, yield: 63.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.21 (s, 1H), 8.74-8.73 (s, 1H), 8.64 (s, 1H) , 8.27 (s, 1H), 3.64-3.62 (m, 4H), 3.20 (s, 3H), 2.84-2.83 (d, 3H), 1.96 (m , 1H), 1.79-1.77 (m, 2H), 1.24 (s, 3H).

Molecular formula: C ₁₈ H ₂₁ N ₅ O ₃ , Molecular weight: 355.40 LC-MS (Pos, m/z) = 356.26 [M+H] ⁺ .

Example 79

Synthesis of 4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 132)

Intermediate 6-chloro-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (20.1 g, 60.40 mmol , 1.0 eq.) was dissolved in 1,4-dioxane (600 ml) and H ₂ O (150 ml), after which potassium trifluorovinylborate (12.14 g, 90.6 mmol, 1.5 eq.) was added, cesium carbonate (58 g, 181.2 mmol, 3.0 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (4.4 g, 6.04 mmol, 1.0 eq.) and left to react at 100°C for 8 hours in a nitrogen atmosphere. The completion of the reaction was determined by LC-MS. Then, water (20 ml) was added to the reaction solution, which was extracted with dichloromethane (30 ml×3). The organic phase was washed with water (10 ml*3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure; the crude product was purified by silica gel column chromatography (DCM:MeOH=50:1) to give the product (14.63 g, yield: 74%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.03 (s, 1H), 8.64 (s, 1H), 7.56 (s, 1H), 6.89 -6.96 (m, 1H), 6.15-6.19 (m, 1H), 5.39-5.42 (m, 1H), 3.61-3.64 (m, 4H), 3 .19 (s, 3H), 1.77-1.93 (m, 4H), 1.21 (s, 3H).

Molecular formula: C ₁₈ H ₂₀ N ₄ O ₂ , Molecular weight: 324.38 LC-MS (Pos, m/z) = 325.16 [M+H] ⁺ .

Example 80

Synthesis of 6-(1-hydroxy-2-methylpropyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridin-3 -carbonitrile (Compound 134)

Step 1: Synthesis of 6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Sodium periodate (4.00 g, 18.70 mmol, 2.0 eq.) was dissolved in water (10 ml), after which tetrahydrofuran (100 ml) was added, cooled to 0° C. in an ice bath, then 6- (1,2-dihydroxyethyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (3, 18 g, 9.35 mmol, 1.0 eq) and stirred at room temperature for 3 hours. Upon completion of the reaction, as shown by TLC, the mixture was extracted with dichloromethane (20 ml × 3). The organic phase was separated, washed with brine (20 ml × 2), dried with anhydrous Na ₂ SO ₄ (0.8 g), filtered with suction and concentrated under reduced pressure, obtaining the product as a yellow solid (1.50 g, yield: 52.0%).

Step 2: Synthesis of 6-(1-hydroxy-2-methylpropyl)-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7- naphthyridine-3-carbonitrile

6-formyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (968.0 mg, 3 ,14 mmol, 1.0 eq.) was dissolved in freshly distilled tetrahydrofuran (100 ml), cooled to -30°C under nitrogen in a dry ice/ethanol bath, and then a solution of isopropylmagnesium chloride in tetrahydrofuran (2 mol/ L, 7.85 ml, 15.70 mmol, 5.0 eq.). Upon completion of the dropwise addition, the mixture was stirred at -30°C and left to react for 30 minutes. The completion of the reaction was determined by TLC. The reaction solution was quenched with saturated aqueous NH ₄ Cl (30 ml) and extracted with dichloromethane (30 ml×3). The organic phase was separated, washed with brine (30 ml × 2), dried with anhydrous Na ₂ SO ₄ (0.8 g), filtered with suction and concentrated under reduced pressure; the crude product was purified by silica gel column chromatography (DCM:MeOH=40:1-20:1) to give a yellow solid (564.0 mg, yield: 50.9%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.96 (s, 1H), 8.59 (s, 1H), 7.71 (s, 1H), 5.36 -5.35 (d, 1H), 4.47-4.45 (t, 1H), 3.66-3.63 (t, 4H), 2.09-2.01 (m, 1H), 1 .63 (s, 4H), 0.90-0.88 (d, 3H), 0.75-0.73 (d, 3H), 0.45 (m, 4H).

Molecular formula: C ₂₀ H ₂₄ N ₄ O ₂ , Molecular weight: 352.44 LC-MS (Pos, m/z) = 353.26 [M+H] ⁺ .

Example 81

Synthesis of N-(2-aminoethyl)-3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxamide hydrochloride ( connection 138)

Step 1: Synthesis of tert-butyl (2-(3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine)-6- formylamino)ethyl)carbamate

Intermediate 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (200 mg, 0.58 mmol, 1.0 eq.), HATU (333 mg, 0.88 mmol, 1.5 eq.) and DIPEA (376 mg, 1.76 mmol, 3.0 eq.) were dissolved in DMAA (2 ml), stirred at room temperature for 30 minutes, then added tert-butyl-(2-aminoethyl)carbamate (281 mg, 1.76 mmol, 2.0 eq.) and left to react at room temperature for 1 hour. The completion of the reaction was determined by LC-MS. Then, water (10 ml) was added to the reaction solution, which was extracted with dichloromethane (10 ml×3). The organic phase was washed with water (10 ml×3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=30:1) to give the product (220 mg, yield: 78.3%).

Step 2: Synthesis of N-(2-aminoethyl)-3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6 hydrochloride -carboxamide

Hydrochloride of Compound 138 formylamino)ethyl)carbamate (220 mg, 0.45 mmol, 1.0 eq.) was dissolved in methanol (3 ml), then a solution of hydrogen chloride in ethanol (25%, 2 ml) was added and left to react at room temperature for 2 hours. The completion of the reaction was determined by TLC. A solid precipitated out of the solution and the resulting mixture was filtered. The filter cake was dried to give the product (150 mg, yield: 79%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.28 (s, 1H), 9.01-9.03 (m, 1H), 8.68 (s, 1H) , 8.29 (s, 1H), 7.91 (s, 3H), 3.55-3.64 (m, 6H), 3.21 (s, 3H), 3.00-3.02 (m , 2H), 1.94-1.97 (d, 2H), 1.73-1.80 (d, 2H), 1.24 (s, 3H).

Molecular formula: C ₁₉ H ₂₄ N ₆ O ₃ , Molecular weight: 384.44 LC-MS (Pos, m/z) = 385.19 [M+H] ⁺ .

Example 82

Synthesis of 3-cyano-N-(2-(dimethylamino)ethyl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6- carboxamide (Compound 139)

Intermediate 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (200 mg, 0.58 mmol, 1.0 eq.), HATU (333 mg, 0.88 mmol, 1.5 eq.) and DIPEA (226 mg, 1.76 mmol, 3.0 eq.) were dissolved in DMAA (2 ml), stirred at room temperature for 30 minutes, after which was added N,N-dimethylethylenediamine (103 mg, 1.16 mmol, 2.0 eq.) and left to react at room temperature for 1 hour. The completion of the reaction was determined by LC-MS. Water (10 ml) was added to the reaction solution, and the reaction solution was extracted with dichloromethane (10 ml×3). The organic phase was washed with water (10 ml×3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure; the crude product was purified by silica gel column chromatography (DCM:MeOH=20:1) to give the product (86 mg, yield: 36%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.20 (s, 1H), 8.79 (s, 1H), 8.66 (s, 1H), 8.28 (s, 1H), 3.62-3.64 (d, 4H), 3.50-3.51 (d, 2H), 3.21 (s, 3H), 2.76 (s, 2H), 2.44 (s, 6H), 1.94-1.97 (d, 2H), 1.74-1.81 (m, 2H), 1.25 (s, 3H).

Molecular formula: C ₂₁ H ₂₈ N ₆ O ₃ , Molecular weight: 412.49 LC-MS (Pos, m/z) = 413.22 [M+H] ⁺ .

Example 83

Synthesis of 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-N-(2-(pyrrolidin-1-yl)ethyl)-1,2-dihydro-1,7- naphthyridine-6-carboxamide (Compound 140)

Intermediate 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (200 mg, 0.58 mmol, 1.0 eq.), HATU (333 mg, 0.88 mmol, 1.5 eq.) and DIPEA (226 mg, 1.76 mmol, 3.0 eq.) were dissolved in DMAA (2 ml), stirred at room temperature for 30 minutes, after which 2-(pyrrolidin-1-yl)ethan-1-amine (134 mg, 1.16 mmol, 2.0 eq.) was added and left to react at room temperature for 1 hour. The completion of the reaction was determined by LC-MS. Then, water (10 ml) was added to the reaction solution, which was extracted with dichloromethane (10 ml×3). The organic phase was washed with water (10 ml×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure; the crude product was purified by silica gel column chromatography (DCM:MeOH=20:1) to give the product (106 mg, yield: 41%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.26 (s, 1H), 9.08-9.11 (m, 1H), 8.67 (s, 1H) , 8.30 (s, 1H), 3.63-3.64 (d, 8H), 3.06-3.20 (m, 6H), 1.73-1.98 (m, 9H), 1 .25 (s, 3H).

Molecular formula: C ₂₃ H ₃₀ N ₆ O ₃ , Molecular weight: 438.53 LC-MS (Pos, m/z) = 439.24 [M+H] ⁺ .

Example 84

Synthesis of 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-N-((1-methylpiperidin-4-yl)methyl)-2-oxo-1,2-dihydro-1,7 trifluoroacetate -naphthyridine-6-carboxamide (Compound 141)

Trifluoroacetate compound 141

Intermediate 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (200 mg, 0.58 mmol, 1.0 eq.) was dissolved in anhydrous N,N-dimethylacetamide (2 ml), after which DIPEA (226.3 mg, 1.75 mmol, 3.0 eq.) and HATU (333.1 mg, 0.0 eq.) were added. 88 mmol, 1.5 eq.), stirred at room temperature for 0.5-1 hour, after which (1-methylpiperidin-4-yl)methanamine (150 mg, 1.17 mmol, 2.0 eq. ) and left to react for 1 hour at room temperature. After starting materials were still detectable by LC-MS, (1-methylpiperidin-4-yl)methanamine (150 mg, 1.17 mmol, 2.0 eq) was added again and allowed to react again for 2 hours. The mixture was purified by preparative HPLC (0.1% aqueous trifluoroacetic acid:acetonitrile = 70:30) to give the product (68.8 mg, yield: 20.7%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.22 (s, 1H), 9.00-8.99 (s, 1H), 8.95-8.94 ( s, 1H), 8.65 (s, 1H), 8.28 (s, 1H), 3.63-3.62 (m, 4H), 3.43-3.40 (m, 2H), 3 .23 (s, 3H), 2.95-2.83 (m, 2H), 2.75-2.74 (m, 2H), 1.97-1.94 (m, 2H), 1.85 -1.80 (m, 2H), 1.78-1.75 (m, 3H), 1.24 (s, 3H).

Molecular formula: C ₂₄ H ₃₂ N ₆ O ₃ , Molecular weight: 452.56 LC-MS (Pos, m/z) = 453.45 [M+H] ⁺ .

Example 85

Synthesis of trifluoroacetate 6-carboxamide

(Compound 142)

Compound 142 trifluoroacetate Intermediate 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acids (200 mg, 0. 58 mmol, 1.0 eq.) was dissolved in anhydrous N,N-dimethylacetamide (2 ml), after which DIPEA (226.3 mg, 1.75 mmol, 3.0 eq.) and HATU (333.1 mg , 0.88 mmol, 1.5 eq.), stirred at room temperature for 0 to 1 hour, and then 1-methylazetidine-3-amine (100.6 mg, 1.17 mmol, 2.0 eq.) and left to react at room temperature for 12 hours. The crude product was purified by preparative HPLC (0.1% aqueous trifluoroacetic acid: acetonitrile = 70:30) to give the product (113.13 mg, yield: 37.1%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.27 (s, 1H), 9.59-9.53 (s, 2H), 8.68 (s, 1H) , 8.27 (s, 1H), 4.90-4.86 (m, 1H), 4.45 (m, 2H), 4.16 (m, 2H), 3.63-3.62 (m , 4H), 3.20 (s, 3H), 2.91 (s, 3H), 1.96-1.93 (m, 2H), 1.79-1.72 (m, 2H), 1, 24 (s, 3H).

Molecular formula: C ₂₁ H ₂₆ N ₆ O ₃ , Molecular weight: 410.48 LC-MS (Pos, m/z) = 411.40 [M+H] ⁺ .

Example 86

Synthesis of 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-N-(1-methylpiperidin-4-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6 -carboxamide (Compound 143)

Starting material, 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (200 mg, 0.58 mmol, 1.0 eq.), was dissolved in N,N-dimethylformamide (2 ml), after which N,N-diisopropylethylamine (226 mg, 1.75 mmol, 3.0 eq.) and HATU (333 mg, 0.87 mmol, 1.5 eq.), left to react at room temperature for 1 hour, then 1-methylpiperidine-4-amine (67 mg, 0.58 mmol, 1.0 eq.) was added and left to react at room temperature for 2 hours. The completion of the reaction was determined by LC-MS. The reaction solution was purified by preparative HPLC (0.1% aqueous trifluoroacetic acid: acetonitrile = 70:30) to give the product (49 mg, yield: 19%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.22 (s, 1H), 9.37 (s, 1H), 8.90 (m, 1H), 8.67 (s, 1H), 8.27 (s, 1H), 4.02-4.04 (m, 1H), 3.62-3.64 (m, 4H), 3.46-3.48 (m , 2H), 3.21 (s, 3H), 3.09 (m, 1H), 2.78 (s, 3H), 1.88-2.01 (m, 6H), 1.73-1, 80 (m, 2H), 1.24 (s, 3H).

Molecular formula: C ₂₃ H ₃₀ N ₆ O ₃ , Molecular weight: 438.53 LC-MS (Pos, m/z) = 439.37 [M+H] ⁺ .

Example 87

Synthesis of 3-cyano-N-(2,3-dihydroxypropyl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxamide (Compound 144)

Intermediate 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (200 mg, 0.58 mmol, 1.0 eq.) was dissolved in anhydrous N,N-dimethylacetamide (2 ml), after which DIPEA (226.3 mg, 1.75 mmol, 3.0 eq.) and HATU (333.1 mg, 0.0 eq.) were added. 88 mmol, 1.5 eq.), stirred at room temperature for 0.5 to 1 hour, then 3-aminopropane-1,2-diol (106.4 mg, 1.17 mmol, 2 0 eq.) and left to react at room temperature for 12 hours. The crude product was purified by preparative HPLC (0.1% aqueous trifluoroacetic acid: acetonitrile = 70:30) and lyophilized to give a sample (93.79 mg). The sample was dissolved in water, adjusted to pH 8 with aqueous sodium bicarbonate, extracted with n-butanol (20 ml×5), and the organic phase was concentrated to give the product (47.2 mg, yield: 19.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 8.42-8.39 (s, 1H), 8.36 (s, 1H), 8.08 (s, 1H) , 4.96 (s, 1H), 4.66 (s, 1H), 3.49 (m, 1H), 3.47-3.45 (m, 6H), 3.25-3.23 (m , 2H), 3.19 (s, 3H), 1.91-1.88 (m, 2H), 1.75-1.70 (m, 2H), 1.23 (s, 3H).

Molecular formula: C ₂₀ H ₂₅ N ₅ O ₅ , Molecular weight: 415.45 LC-MS (Neg, m/z) = 414.34 [M-H] ^- .

Example 88

Synthesis of 4-(4-methoxy-4-methylpiperidin-1-yl)-6-(2-methoxyethoxy)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 145)

Step 1: Synthesis of methyl 2-(2-methoxyethoxy)-5-nitroisonicotinate

The crude material, ethylene glycol monomethyl ether (3.5 g, 46.17 mmol, 1.0 eq.), was dissolved in tetrahydrofuran (50 ml), cooled to 0°C, after which sodium hydride (3.7 g, 92.34 mmol, 2.0 eq.). The reaction proceeded for 1 hour, after which was added methyl-2-chloro-5-nitroisonicotinate (10.0 g, 46.17 mmol, 1.0 eq.). The completion of the reaction was determined by TLC (PE:EA = 5:1). The reaction solution was poured into ice water (100 ml) and quenched. The aqueous phase was extracted with ethyl acetate (100 ml×2), the organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (PE:EA=5:1) to give the product as a pale yellow oil (1.8 g, yield: 15%).

Step 2: Synthesis of methyl 5-amino-2-(2-methoxyethoxy)isonicotinate

The intermediate methyl 2-(2-methoxyethoxy)-5-nitroisonicotinate (1.8 g, 7.02 mmol, 1.0 eq.) was dissolved in methanol (10 mL) followed by the addition of 10% palladium on carbon (500 mg), were supplied with hydrogen and left to react overnight at room temperature. The completion of the reaction was determined by TLC (PE:EA = 3:1). The reaction solution was filtered and concentrated, and the crude product was purified by silica gel column chromatography (PE:EA=5:1) to give the product as a pale yellow solid (1.2 g, yield: 75%).

Step 3: Synthesis of methyl 5-(2-cyanoacetamido)-2-(2-methoxyethoxy)isonicotinate

Intermediate methyl 5-amino-2-(2-methoxyethoxy)isonicotinate (1.2 g, 5.3 mmol, 1.0 eq.) and cyanoacetic acid (901 mg, 10.6 mmol, 2.0 eq.) was dissolved in dichloromethane (20 ml), after which EDCI (3.04 g, 15.9 mmol, 3.0 eq.) was added and left to react at room temperature for 2 hours. The completion of the reaction was determined by LC-MS. The reaction solution was poured into ice water (30 ml) and quenched. The aqueous phase was extracted with dichloromethane (30 ml×2), the organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was suspended in methyl tert-butyl ether to give the product as a pale yellow solid (1.2 g, yield: 77%).

Step 4: Synthesis of 4-hydroxy-6-(2-methoxyethoxy)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

The intermediate methyl 5-(2-cyanoacetamido)-2-(2-methoxyethoxy)isonicotinate (1.2 g, 4.09 mmol, 1.0 eq.) was dissolved in THF (20 mL), after which sodium hydride ( 327 mg, 8.18 mmol, 2.0 eq.), then heated to 80°C and left to react for 4 hours. The completion of the reaction was determined by LC-MS. The reaction solution was cooled to about 0° C., adjusted to pH 2 with a 2 mol/L aqueous hydrochloric acid solution so that no solid precipitated, and then filtered. The filter cake was dried at 50° C. under normal pressure to give the product as a yellow solid (800 mg, yield: 75%).

Step 5: Synthesis of 4-chloro-6-(2-methoxyethoxy)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 4-hydroxy-6-(2-methoxyethoxy)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (800 mg, 3.06 mmol, 1.0 eq) was dissolved in oxychloride phosphorus (8 ml), heated to 100°C and left to react for 1 hour. The completion of the reaction was determined by LC-MS. The reaction solution was poured into ice water (20 ml) and quenched. The aqueous phase was extracted with dichloromethane (30 ml×3), the organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was suspended in methyl tert-butyl ether to give the product as a pale yellow solid (180 mg, yield: 21%).

Step 6: Synthesis of 4-(4-methoxy-4-methylpiperidin-1-yl)-6-(2-methoxyethoxy)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

The intermediate 4-chloro-6-(2-methoxyethoxy)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (180 mg, 0.64 mmol, 1.0 eq) was dissolved in N ,N-dimethylformamide (2 mL) followed by the addition of N,N-diisopropylethylamine (332 mg, 2.58 mmol, 4.0 eq.) and 4-methoxy-4-methylpiperidine (110 mg, 0.97 mmol, 1 0 eq.), heated to 80°C and then left to react for two hours. The completion of the reaction was determined by LC-MS. The reaction solution was poured into ice water (20 ml) and quenched. The aqueous phase was extracted with dichloromethane (30 ml×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (DCM:MeOH=15:1) to give the product as a pale yellow oil (60 mg, yield: 25%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.78 (s, 1H), 8.28 (s, 1H), 6.94 (s, 1H), 4.37 (m, 2H), 3.67 (m, 2H), 3.56-3.58 (m, 4H), 3.30 (s, 3H), 3.18 (s, 3H), 1.88- 1.91 (m, 2H), 1.75-1.80 (m, 2H), 1.21 (s, 3H).

Molecular formula: C ₁₉ H ₂₄ N ₄ O ₄ , Molecular weight: 372.43 LC-MS (Pos, m/z) = 373.3 [M+H] ⁺ .

Example 89

Synthesis of 3-cyano-N,N-dimethyl-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-6-carboxamide (Compound 149)

3-cyano-2-oxo-4-(6-aza-spiro[2.5]octan-6-yl)-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (256.4 mg, 0.790 mmol, 1.0 eq.) was dissolved in DMF (20 ml), cooled to 0°C in an ice bath, after which HATU (450.8 mg, 1.186 mmol, 1.5 eq.) was added and then DIPEA ( 613.0 mg, 4.743 mmol, 6.0 eq.) and dimethylamine hydrochloride (193.3 mg, 2.371 mmol, 3.0 eq.). Upon completion of the addition, the mixture was stirred at room temperature for 30 minutes. The completion of the reaction was determined by TLC. Water (100 ml) was added to the mixture and extracted with ethyl acetate (20 ml×3). The organic phases were combined, washed with brine (10 ml×4), dried with anhydrous sodium sulfate, suction filtered, concentrated under reduced pressure and purified by preparative thin layer chromatography (DCM:MeOH = 8:1) to give the product as a yellow solid substances (91.0 mg, yield: 32.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.19 (s, 1H), 8.61 (s, 1H), 7.89 (s, 1H), 3.68 -3.65 (t, 4H), 3.06 (s, 3H), 3.02 (s, 3H), 1.61 (s, 4H), 0.44 (s, 4H).

Molecular formula: C ₁₉ H ₂₁ N ₅ O ₂ , Molecular weight: 351.41 LC-MS (Pos, m/z)=352.20 [M+H] ⁺ .

Example 90

Synthesis of 4-(4-(2-hydroxyethyl)piperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 153)

Intermediate 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (60 mg, 0.29 mmol, 1.0 eq.) was dissolved in N,N-dimethylformamide (1 ml) , and N,N-diisopropylethylamine (112 mg, 0.87 mmol, 3.0 eq.) was added to the reaction mixture, followed by 2-(piperidin-4-yl)ethan-1-ol (38 mg, 0.29 mmol, 1.0 eq.). The temperature was raised to 80°C, and the reaction was carried out for 2 hours. The mixture was cooled to room temperature, resulting in a solid precipitate, which was then filtered. The filter cake was rinsed successively with THF (1 ml) and petroleum ether (1 ml) and dried at 45° C. to give a yellow solid (16 mg, yield: 18%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.98 (s, 1H), 8.65 (s, 1H), 8.33-8.35 (d, 1H) , 7.58-7.80(d, 1H), 4.40-4.42(m, 1H), 3.83-3.86(m, 2H), 3.48-3.53(m, 2H), 3.37-3.43(m, 2H), 1.84-1.87(m, 2H), 1.74-1.80(m, 1H), 1.45-1.50( m, 4H).

Molecular formula: C ₁₆ H ₁₈ N ₄ O ₂ , Molecular weight: 298.35 LC-MS (Neg, m/z)=297.15[MH] ^- .

Example 91

Synthesis of (R)-4-(3-(1H-1,2,4-triazol-1-yl)pyrrolidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3- carbonitrile (Compound 154)

Step 1: Synthesis of tert-butyl (R)-3-(1H-1,2,4-triazol-1-yl)pyrrolidine-1-carboxylate

The starting material, tert-butyl-(S)-3-hydroxypyrrolidine-1-carboxylate (1.0 g, 5.34 mmol, 1.0 eq.), was dissolved in anhydrous tetrahydrofuran (20 ml) and the reaction mixture was added 1H-1,2,4-triazole (552.7 mg, 8.01 mmol, 1.5 eq.) and triphenylphosphine (2.8 g, 10.68 mmol, 2.0 eq.). Upon completion of the addition, the temperature was lowered to 0° C. and diethyl azodicarboxylate (1.86 g, 10.68 mmol, 2.0 eq.) was added dropwise. Upon completion of the addition, the reaction was carried out at room temperature for 12 hours. The completion of the reaction was determined by TLC. Saturated aqueous sodium carbonate solution (10 ml) was added to the resulting solution, the solution mixture was extracted with ethyl acetate (50 ml×2), and the layers were separated. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (EA:PE=1:10-1:2) to give the product (1.05 g, yield: 83.3%).

Step 2: Synthesis of (R)-1-(pyrrolidin-3-yl)-1H-1,2,4-triazole hydrochloride

The intermediate tert-butyl-(R)-3-(1H-1,2,4-triazol-1-yl)pyrrolidine-1-carboxylate (1.05 g, 4.41 mmol, 1.0 eq) was dissolved in anhydrous methanol (4 ml), after which a 30% solution of hydrogen chloride in ethanol (10 ml) was added and left to react at room temperature for 2 hours until a white solid precipitated. The completion of the reaction was determined by TLC. The reaction solution was filtered, the filter cake was rinsed with ethyl acetate and dried to give the product (770 mg, yield: 100%).

Step 3: Synthesis of (R)-4-(3-(1H-1,2,4-triazol-1-yl)pyrrolidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine -3-carbonitrile

The intermediate 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (200 mg, 0.97 mmol, 1.0 eq.) was dissolved in DMF (2 ml), and in the reaction (R)-1-(pyrrolidin-3-yl)-1H-1,2,4-triazole hydrochloride (238 mg, 1.36 mmol, 1.4 eq) and DIPEA (753.7 mg, 5 .84 mmol, 6.0 eq.). Upon completion of the addition, the temperature was raised to 80°C, and the reaction was carried out for 2 hours. The completion of the reaction was determined by LC-MS. The reaction solution was cooled to room temperature and purified by preparative HPLC (0.1% aqueous trifluoroacetic acid: acetonitrile = 70:30) to give the product (98.6 mg, yield: 33%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.67 (s, 1H), 8.69 (s, 1H), 8.62 (s, 1H), 8.28 -8.27 (d, 1H), 8.03 (s, 1H), 7.98-7.96 (s, 1H), 5.33-5.28 (m, 1H), 4.61-4 .57 (m, 1H), 4.35-4.29 (m, 2H), 4.23-4.17 (m, 1H), 2.56-2.53 (m, 1H), 2.48 -2.47 (m, 1H).

Molecular formula: C ₁₅ H ₁₃ N ₇ O, Molecular weight: 307.21 LC-MS (Pos, m/z)=307.98 [M+H] ⁺ .

Example 92

Synthesis of 4-(R)-3-(1Н-pyrazol-1-yl)pyrrolidin-1-yl)-6-(1-hydroxyethyl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3 -carbonitrile (Compound 156)

Step 1: Synthesis of (R)-1-(pyrrolidin-3-yl)-1H-pyrazole hydrochloride

The starting material, tert-butyl-(R)-3-(1H-pyrazol-1-yl)pyrrolidine-1-carboxylate (2.7 g, 11.37 mmol), was dissolved in anhydrous methanol (4 ml), and in the reaction mixture was added a 30% solution of hydrogen chloride in ethanol (10 ml). Upon completion of the addition, the reaction was carried out at room temperature for 2 hours. The completion of the reaction was determined by TLC. The mixture was concentrated under reduced pressure to give an oily product (3.29 g crude).

Step 2: Synthesis of (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3 -carbonitrile

The intermediate 4-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (2.27 g, 9.46 mmol, 1.0 eq.) was dissolved in DMF (8 ml), and (R)-1-(pyrrolidin-3-yl)-1H-pyrazole hydrochloride (3.29 g crude) and DIPEA (7.3 g, 56.76 mmol, 6.0 eq.) were added to the reaction mixture. Upon completion of the addition, the temperature was raised to 80°C, and the reaction was carried out for 2 hours.

The completion of the reaction was determined by the LC-MS method, and the temperature was lowered to room temperature. The reaction solution was poured into water (50 ml), stirred for half an hour, filtered. The filter cake was rinsed with water, then suspended in methyl tert-butyl ether and filtered; the filter cake was dried to give the product (2.35 g, yield from two synthetic steps: 73.4%).

Step 3: Synthesis of (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-2-oxo-6-vinyl-1,2-dihydro-1,7-naphthyridine-3 -carbonitrile

Intermediate (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-chloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile ( 2.3 g, 6.75 mmol, 1.0 eq.), cesium carbonate (6.6 g, 20.25 mmol, 3.0 eq.) and potassium vinyltrifluoroborate (1.3 g, 10.12 mmol, 1.5 eq) was dissolved in a solvent mixture of 1,4-dioxane (50 ml) and water (50 ml). The air in the system was displaced with nitrogen three times and then [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (493 mg, 0.68 mmol, 0.1 eq.) was added to the reaction mixture. The air in the system was purged with nitrogen three times, the mixture was refluxed and left to react for 12 hours. The completion of the reaction was determined by TLC, and the temperature was lowered to room temperature. Ethyl acetate (50 ml) and water (10 ml) were added, stirred for 10 minutes and filtered. The filter cake was rinsed with ethyl acetate and the liquid was separated. The aqueous phase was extracted with dichloromethane (50 ml×3), the organic phases were combined, dried over anhydrous sodium sulfate and filtered. The mother liquor was concentrated under reduced pressure, suspended in methyl tert-butyl ether, which resulted in the precipitation of a pale yellow solid, and then filtered. The filter cake was dried to give the product (1.45 g, yield: 65.9%).

Step 4: Synthesis of 4-((R)-3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-(1,2-dihydroxyethyl)-2-oxo-1,2-dihydro-1 ,7-naphthyridine-3-carbonitrile

Intermediate (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-2-oxo-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile ( 453 mg, 1.36 mmol, 1.0 eq.) was dissolved in a mixture of solvents of tert-butanol (7 ml) and water (7 ml), after which methanesulfonamide (129.5 mg, 1.36 mmol, 1.0 eq.) and AD-mix-β (5.4 g). Upon completion of the addition, the reaction in the resulting mixture was left to proceed at room temperature for 12 hours. The completion of the reaction was determined by LC-MS. The reaction solution was directly introduced into the next step.

Step 5: Synthesis of (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-formyl-2-oxo-1,2-dihydro-1,7-naphthyridine-3 -carbonitrile

Sodium periodate (582.6 mg, 2.72 mmol, 2.0 eq.) and tetrahydrofuran (5 ml) were added to the reaction solution obtained in the previous step. After completion of the addition, the reaction was carried out for 4 hours at room temperature, and the completion of the reaction was determined by the LC-MS method. Water (20 ml) was added and the liquid separated. The aqueous phase was extracted with dichloromethane (50 ml×4), the organic phases were combined, dried and filtered. The filtrate was concentrated under reduced pressure to give a pale yellow solid. The solid was suspended in methyl tert-butyl ether, filtered and the filter cake dried to give the product (215 mg, yield: 47.2%).

Step 6: Synthesis of 4-(R)-3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-(1-hydroxyethyl)-2-oxo-1,2-dihydro-1,7- naphthyridine-3-carbonitrile

Intermediate (R)-4-(3-(1H-pyrazol-1-yl)pyrrolidin-1-yl)-6-formyl-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile ( 215 mg, 0.64 mmol, 1.0 eq.) was dissolved in anhydrous tetrahydrofuran (3 ml), cooled to -10 ~ 0°C under nitrogen atmosphere, then a solution of methylmagnesium chloride in tetrahydrofuran (3 mol/l, 0 .32 ml, 0.96 mmol, 1.5 eq.). Upon completion of the addition, the reaction was carried out for 12 hours at room temperature. The completion of the reaction was determined by LC-MS. The pH of the reaction solution was adjusted to approximately 8 by dropwise addition of a saturated aqueous ammonium chloride solution, and extracted with dichloromethane (50 ml×4). The organic phases were combined, dried, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (MeOH:DCM = 1:100 to 1:40) to give the product (44.6 mg, yield: 19.8%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.59 (s, 1H), 8.54 (s, 1H), 8.00 (s, 1H), 7.90 -7.89(d, 1H), 7.58-7.56(d, 1H), 7.51(d, 1H), 6.31-6.30(d, 1H), 6.29(d , 1H), 5.46-5.45(m, 1H), 5.20(d, 1H), 4.79-4.73(m, 1H), 4.57-4.50(m, 1H ), 4.37-4.20 (m, 1H), 2.48 (m, 2H), 1.38-1.37 (d, 3H).

Molecular formula: C ₁₈ H ₁₈ N ₆ O ₂ , Molecular weight: 350.38 LC-MS (Pos, m/z)=351.22[M+H] ⁺ .

Example 93

Synthesis of 4-(4-methoxy-4-methylpiperidin-1-yl)-6-methyl-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 158)

Intermediate 6-chloro-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (1.3 g, 3.91 mmol , 1.0 eq.), cesium carbonate (3.8 g, 11.73 mmol, 3.0 eq.) and trimethylcyclotriborane (50% in THF, 3.9 g, 15.62 mmol, 4.0 eq. ) was dissolved in 1,4-dioxane (20 ml). Upon completion of the addition, the air in the system was expelled three times with nitrogen, and then [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (286 mg, 0.39 mmol, 0.1 eq.) was added to the reaction mixture. Upon completion of the addition, the air in the system was purged with nitrogen three times and the mixture was refluxed for 12 hours. When starting materials were still present in the system by TLC, additional trimethylcyclotriborane (50% solution in THF, 3.9 g, 15.62 mmol, 4.0 eq.) and dichloride [1,1'-bis (diphenylphosphino)ferrocene]palladium (286 mg, 0.39 mmol, 0.1 eq.) and boiling was continued for 4 hours. When no starting material remained in the system by TLC, the system was cooled to room temperature, after which water (50 ml) and dichloromethane (100 ml) were added, stirred for 5 minutes, which led to the appearance of a solid precipitate, which was filtered. The filter cake was rinsed with dichloromethane. The liquid was separated, the aqueous phase was extracted with dichloromethane (100 ml×3), the organic phases were combined, dried over anhydrous sodium sulfate and filtered. The mother liquor was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (MeOH:DCM=1:100-1:50) to give the product (309.9 mg, yield: 25.4%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.88 (s, 1H), 8.55 (s, 1H), 7.42 (s, 1H), 3.61 -3.59 (m, 4H), 3.19 (s, 3H), 2.53 (s, 3H), 1.92-1.89 (m, 2H), 1.82-1.75 (m , 2H), 1.22(s, 3H).

Molecular formula: C ₁₇ H ₂₀ N ₄ O ₂ , Molecular weight: 312.37 LC-MS (Pos, m/z)=313.25 [M+H] ⁺ .

Example 94

Synthesis of 6-(cyclopropyl(hydroxy)methyl)-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (Compound 160)

Step 1: Synthesis of 6-chloro-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 4,6-dichloro-2-oxo-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (8.0 g, 33.3 mmol, 1.0 eq) was dissolved in DMF (100 ml) , after which the starting material, 7-aza-spiro[3,5]nonane hydrochloride (6.5 g, 40.0 mmol, 1.2 eq.), and DIPEA (17.2 g, 133.2 mmol , 4.0 eq.), and left to react at 80°C for half an hour. The completion of the reaction was determined by TLC. The reaction solution was concentrated, after which water and ethyl acetate (40 ml/40 ml) were added, stirred overnight and filtered to give a yellow solid (10.0 g, yield: 91.5%).

Step 2: Synthesis of 2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 6-chloro-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (5.0 g, 15.2 mmol, 1.0 eq.) was dissolved in 1,4-dioxane (50 ml). Starting materials, potassium vinylfluoroborate (6.1 g, 45.6 mmol, 3.0 eq.), cesium carbonate (14.8 g, 45.6 mmol, 3.0 eq.) and dichloride [1,1'- bis(diphenylphosphino)ferrocene]palladium (1.1 g, 15.2 mmol, 1.0 eq.) was dissolved in water (10 ml) and added to the reaction solution obtained as above. The reaction mixture was refluxed overnight under nitrogen atmosphere. The completion of the reaction was determined by LC-MS. The reaction solution was concentrated and extracted with EA (3×100 ml). The organic phase was dried with anhydrous sodium sulfate, filtered with suction and concentrated under reduced pressure to give the product (4.5 g, yield: 92.5%).

Step 3: Synthesis of 6-formyl-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridine-3-carbonitrile

Intermediate 2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-6-vinyl-1,2-dihydro-1,7-naphthyridine-3-carbonitrile (2.0 g, 6.2 mmol, 1.0 eq.) was dissolved in tert-butanol (50 ml) and water (40 ml), after which Ad-mix-β (20 g, 10 eq.) and methanesulfonamide were added successively at 0°C. (1.2 g, 12.5 mmol, 2.0 eq.) and stirred at room temperature for 48 hours. The completion of the reaction was determined by LC-MS. Sodium periodate (4.0 g, 18.6 mmol, 3.0 eq.) was dissolved in water (10 ml) at 0° C. and added dropwise to the above reaction solution. The reaction solution was stirred at room temperature for 12 hours. The completion of the reaction was determined by LC-MS. The reaction solution was extracted with EA (3×100 ml). The organic phase was dried with anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=40:1) to give the product as a yellow solid (1.0 g, yield: 50.4%).

Step 4: Synthesis of 6-(cyclopropyl(hydroxy)methyl)-2-oxo-4-(7-aza-spiro[3.5]nonan-7-yl)-1,2-dihydro-1,7-naphthyridin- 3-carbonitrile

Intermediate 6-formyl-2-oxo-4-(7-aza-spiro[3,5]nonan-7-yl)-1,2-digid PC-1,7-naphthyridine-3-carbonitrile (500.0 mg , 1.6 mmol, 1.0 eq.) was dissolved in 2-methyltetrahydrofuran (5 ml), then cyclopropylmagnesium bromide (7.5 ml, 7.75 mmol, 5 eq. ) and stirred at -10°C for 12 hours. The completion of the reaction was determined by LC-MS. The reaction solution was quenched with saturated aqueous ammonium chloride at 0° C. and extracted with EA (3×30 ml). The organic phase was dried with anhydrous sodium sulfate, filtered with suction, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=30:1-20:1) to give the product (80.0 mg, yield: 13.7%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.92 (br, 1H), 8.58 (s, 1H), 7.69 (s, 1H), 5.41 (d, 1H), 4.23-4.26 (m, 1H), 3.54 (s, 4H), 1.80-1.99 (m, 9H), 1.11-1.24 (m , 2H), 0.42 (s, 4H).

Molecular formula: C ₂₁ H ₂₄ N ₄ O ₂ , Molecular weight: 364.45 LC-MS (m/z)=365 [M+H] ⁺ .

Example 95

Synthesis of 6-(1-hydroxy-2-methylpropan-2-yl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-3 -carbonitrile (Compound 161)

Step 1: Synthesis of ethyl 2-(3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridin-6-yl)- 2-methylpropionate

Intermediate ethyl 2-(4-chloro-3-cyano-2-oxo-1,2-dihydro-1,7-naphthyridin-6-yl)-2-methylpropionate (600.0 mg, 1.87 mmol, 1 0 eq.) was dissolved in DMF (5 ml), after which the starting material, 4-methyl-4-methoxypiperidine hydrochloride (339.4 mg, 2.05 mmol, 1.1 eq.), and DIPEA (1 .4 g, 11.2 mmol, 6.0 eq.) and left to react at 80°C for half an hour. The completion of the reaction was determined by TLC. The reaction solution was concentrated, after which water was added and extracted with EA (3×60 ml). The organic phases were combined, dried with anhydrous sodium sulfate, filtered, and concentrated, and the crude product was purified by silica gel column chromatography (PE:EA=3:2) to give the product (300.0 mg, yield: 38.9%).

Step 2: Synthesis of 6-(1-hydroxy-2-methylpropan-2-yl)-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7- naphthyridine-3-carbonitrile

Intermediate ethyl 2-(3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridin-6-yl)-2-methylpropionate (300.0 mg, 0.73 mmol, 1.0 eq.) was dissolved in anhydrous 2-methyltetrahydrofuran (5 ml), then DIBAL-H (solution with a concentration of 1.5 mol /l in toluene, 2.4 ml, 3.65 mmol, 5.0 eq.), then warmed to room temperature and stirred for 6 hours. The reaction was stopped by adding dropwise water at 0°C, concentrated and extracted with EA (3×60 ml). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (PE:EA = 3:2) to give the product (60.0 mg, yield: 22.2%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 11.88 (s, 1H), 8.63 (s, 1H), 7.53 (s, 1H), 4.65 (t, 1H), 3.59-3.62 (m, 4H), 3.54 (d, 2H), 3.19 (s, 3H), 1.91-1.95 (m, 2H), 1.73-1.78(m, 2H), 1.28(s, 6H), 1.23(s, 3H).

Molecular formula: C ₂₀ H ₂₆ N ₄ O ₃ , Molecular weight: 370.45 LC-MS (Pos, m/z)=371 [M+H] ⁺ .

Example 96

Synthesis of 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-N,N-dimethyl-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxamide (Compound 162)

Intermediate 3-cyano-4-(4-methoxy-4-methylpiperidin-1-yl)-2-oxo-1,2-dihydro-1,7-naphthyridine-6-carboxylic acid (200 mg, 0.58 mmol, 1.0 eq.), HATU (333 mg, 0.88 mmol, 1.5 eq.) and DIPEA (376 mg, 1.76 mmol, 3.0 eq.) were dissolved in DMAA (2 ml), stirred at room temperature for 30 minutes, after which dimethylamine hydrochloride (95 mg, 1.16 mmol, 2.0 eq.) was added and the reaction was left to proceed at room temperature for 1 hour. The completion of the reaction was determined by LC-MS. Water (10 ml) was added and the mixture was extracted with dichloromethane (10 ml×3). The organic phase was washed with water (10 ml×3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (DCM:MeOH=50:1) to give the product (120 mg, yield: 55%).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 12.16 (s, 1H), 8.61 (s, 1H), 7.83 (s, 1H), 3.61 -3.63 (m, 4H), 3.19 (s, 3H), 3.03-3.06 (d, 6H), 1.90-1.93 (d, 2H), 1.71-1 .78 (m, 2H), 1.22 (s, 3H).

Molecular formula: C ₁₉ H ₂₃ N ₅ O ₃ , Molecular weight: 369.18 LC-MS (Pos, m/z)=370.43 [M+H] ⁺ .

Other compounds can be obtained by the methods described above. Characterization data for some other compounds of the present invention are shown in Table 2 below.

The present invention will become more clear after considering the following experimental examples. However, a person skilled in the art should understand that the above content of experimental examples is given to illustrate the invention and should not limit or limit the scope of the invention as disclosed in the appended claims.

Experimental Example 1

Method for assessing the enzymatic properties of PDE9

Test substance: Compounds according to the invention prepared according to the respective embodiments of the invention

1. Experimental materials and equipment Enzyme PDE9A2 (BPS, cat. no.: 60090) 384-well plate (Perkin Elmer, cat. no.: 6007279)

2. Procedure of the experiment

Preparation of Compounds: Compounds were prepared for long-term storage as a 10 mM stock solution of compounds in DMSO (dimethyl sulfoxide). The resulting compound stock solution was diluted 100 times with DMSO to give a working compound stock solution at a concentration of 100 μM, and then the working compound stock solution was diluted 3 times with DMSO to give 8-10 concentration gradients of the diluted compound stock solution (100×).

Incubation with Compounds: An Echo pipetting system was used to pipette a diluted stock solution of a compound into a 384-well plate to pipette very small amounts of liquid. To each compound well was added 200 panoliters of diluted compound stock solution and 10 μl of PDE9A2 enzyme solution. After centrifugation at 1000 rpm for 1 minute, the mixture was incubated for 15 minutes at room temperature. Then 10 µl of the substrate mixture was added. After centrifugation at 1000 rpm for 1 minute, the mixture was incubated with shock for 30 minutes at room temperature. Finally, stop solution was added to stop the reaction in the system. The mixture was incubated with shock for 60 minutes at room temperature. At maximum reading (Max), the compound was displaced by solvent. At the minimum reading (Min), both compound and enzyme solution were replaced with solvent.

Measurements: Fluorescence values (F) at 480 nm/535 nm were determined using a microplate reader.

Calculation: The rate of inhibition was calculated according to the following formula, and the IC ₅₀ values were approximated using GraphPad Prism 5.0 software:

3. The test results are presented in Table 3:

The data presented in Table 3 show that the compounds of the present invention have a very satisfactory PDE9 enzyme inhibitory activity that is of a magnitude suitable for potential clinical use.

Experimental Example 2

Pharmacokinetic evaluation of the compounds according to the invention in experiments on dogs

Test compound: Compounds according to the invention prepared according to the respective embodiments of the invention

1. Animal Administration and Sampling

To prepare the solution, test compound 25 was dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD )) physiological saline solution. A solution of compound 25 was administered intragastrically (orally) to beagle dogs at a dosage of 2.0 mg/kg. Blood sampling was performed at the following time points: 0 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after administration.

To prepare the solution, test compound 25 was dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD) physiological saline. A solution of compound 25 was administered to beagle dogs at a dosage of 1 mg/kg via intravenous bolus injection. Blood samples were taken at the following time points: 0 minutes, 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after administration.

To prepare the solution, test compound 65 was dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD) physiological saline. A solution of compound 65 was administered intragastrically to beagle dogs at a dosage of 2.0 mg/kg. Blood sampling was performed at the following time points: 0 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after administration.

To prepare the solution, test compound 65 was dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD) physiological saline. Compound 65 solution was administered to beagle dogs at a dosage of 1 mg/kg via intravenous bolus injection. Blood samples were taken at the following time points: 0 minutes, 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after administration.

To prepare the solution, test compound 107 was dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD) physiological saline. A solution of compound 107 was administered intragastrically to beagle dogs at a dosage of 1.0 mg/kg. Blood sampling was performed at the following time points: 0 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after administration.

To prepare the solution, test compound 107 was dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD) physiological saline. Compound 107 solution was administered to beagle dogs at a dosage of 1.0 mg/kg via intravenous bolus injection. Blood samples were taken at the following time points: 0 minutes, 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after administration.

To prepare the solution, test compound 158 was dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD) physiological saline. A solution of compound 158 was administered intragastrically to beagle dogs at a dosage of 1.0 mg/kg. Blood sampling was performed at the following time points: 0 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after administration.

To prepare the solution, test compound 158 was dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD) physiological saline. Compound 158 solution was administered to beagle dogs at a dosage of 1.0 mg/kg via intravenous bolus injection. Blood samples were taken at the following time points: 0 minutes, 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after administration.

Approximately 1 ml of blood was taken from a vein of the upper limb.

The collected blood was placed in an anticoagulation tube containing EDTA-K2 (ethylenediaminetetraacetic acid-K2). Blood samples were centrifuged at 1800 g for 5 min at 4° C. to obtain plasma samples. Plasma was prepared within 30 minutes after blood sampling and stored in a freezer at -80°C until testing.

2. Sample analysis method

The test plasma sample was removed from the freezer, which was kept at -80°C, left to warm at room temperature and vortexed for 5 minutes. Twenty (20) µl of the plasma sample was carefully pipetted into a 1.5 ml centrifuge tube, after which 200 µl of the working solution of the internal standard (tolbutamide in methanol) at a concentration of 100 ng/ml was added, mixed, vortexed for 5 minutes and then centrifuged at 12,000 rpm for 5 minutes. 50 μl of the liquid above the pellet was taken and carefully placed with a pipette into 96-well plates, in which water was previously placed in an amount of 150 μl/well; vortexed for 5 minutes and analyzed by LC-MS/MS (liquid chromatography with tandem mass spectrometry). The injection volume of Compound 25, Compound 65, Compound 107 and Compound 158 was 20, 10, 20 and 10 μl, respectively.

3. Method of data processing

Compound concentrations were determined using the Analyst 1.6.3 software supplied by AB SCIEX. Microsoft Excel was used to calculate the mean, standard deviation, coefficient of variation, and other parameters (directly issued by Analyst 1.6.3 and not calculated).

Pharmacokinetic parameters were calculated using Pharsight Phoenix 6.1 NCA software (T _max is the mean).

4. Results

Experimental results are presented in Table 4:

As can be seen from the data above in the Table, the compounds according to the invention have very good pharmacokinetic characteristics.

Experimental Example 3

Evaluation of the Stability of Compounds According to the Invention in Human Liver Microsome

Objective: Evaluation of the stability of the compounds according to the invention in the human liver microsome.

Test substance: Compounds according to the present invention and compound 1-8 discussed in patent WO2017019723A1 having the following structural formula:

The composition of the incubated system is presented in Table 5:

Experiment procedure:

(1) Liver microsomes (20 mg protein/ml) were removed from the freezer kept at -80°C, placed in a thermostatically controlled water bath at 37°C for 3 minutes pre-incubation, and thawed for further use.

(2) The composition of the mixed solution of the incubation system (without compound and β-NADPH) was prepared according to the ratios shown in Table 4 "Composition of the incubation system", and pre-incubated for 2 minutes in a thermostatically controlled water bath at a temperature of 37°C.

(3) Control group (without β-NADPH): 30 µl of water and 30 µl of compound working solution (10 µM, solvent: 1% DMSO aqueous solution) were added to 240 µl of the mixed solution of the incubation system obtained in step (2) , vortexed for 30 seconds and mixed; the total volume of the reaction mixture was 300 µl. The sample was duplicated. The mixture was incubated in a thermostatically controlled water bath at a temperature of 37°C and time was started. Samples were taken at time points 0 minutes and 60 minutes.

(4) Sample group: 70 μl of β-NADPH solution (10 mM) and 70 μl of compound working solution (10 μM) were added to 560 μl of the incubation system mixed solution obtained in step (2). The total volume of the reaction mixture was 700 μl. The mixture was vortexed for 30 seconds, mixed, and the well was duplicated. The mixture was incubated in a thermostatically controlled water bath at a temperature of 37°C, and time was started. Samples were taken at time points 0 minutes, 5 minutes, 10 minutes, 20 minutes, 30 minutes, 60 minutes from the start.

(5) After vortexing for 3 minutes, the sample was centrifuged at 12,000 rpm for 5 minutes.

(6) 50 μl of liquid above the pellet was taken, 150 μl of water was added thereto, vortexed, mixed, and analyzed by LC-MS/MS.

Data analysis:

The half-life (t _1/2 ) and the rate of elimination (CI) were calculated according to the first order dynamics formula below:

Ct=C ₀ *e ^-kt

t _1/2 \u003d ln2 / k \u003d 0.693 / k

Cl _int =V _d *k

V _d =1/(protein content in liver microsomes).

Notes: k is the slope of the logarithmic dependence of the amount of compound remaining on time, and V _d is the observed volume of distribution.

The results are presented in Table 6.

The above results show that the compounds of the present invention have a slower clearance rate from the human liver microsome than the compounds of the prior art.

Experimental Example 4

Pharmacokinetic evaluation of the compounds according to the invention in experiments with rats

Objective: Evaluation of the in vivo pharmacokinetic parameters of compounds in male SD rats (Sprague Dawley rats) (purchased from Beijing Vital River Animal Technology Co., Ltd.) and evaluation of the bioavailability of the compounds.

Animal administration and sampling:

To prepare the solution, test compounds 65, 90, 107, 158 were dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD) physiological saline. A solution of compounds 65, 90, 107, 158 was administered intragastrically to SD rats at a dosage of 5.0 mg/kg. Blood samples were taken at the following time points: 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours after administration.

To prepare the solution, test compounds 65, 90, 107, 158 were dissolved in a mixture of 2% DMSO + 10% PEG400 + 88% (28% HP-β-CD) physiological saline. A solution of compounds 65, 90, 107, 158 was administered to SD rats at a dosage of 1 mg/kg via intravenous bolus injection. Blood sampling was performed at the following time points: 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours after administration.

To prepare the solution, test compound 76 was dissolved in a mixture of 30% DMF + 30% PEG400 + 40% saline. Compound 76 solution was administered intragastrically to SD rats at a dosage of 5.0 mg/kg. Blood samples were taken at the following time points: 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours after administration.

To prepare the solution, test compound 76 was dissolved in a mixture of 30% DMF + 30% PEG400 + 40% saline. Compound 76 solution was administered to SD rats at a dosage of 1 mg/kg via intravenous bolus injection. Blood sampling was performed at the following time points: 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours after administration.

To prepare the solution, test compound 25 was dissolved in a mixture of 5% DMSO + 20% (30% solyutol) + 2% 1M NaOH + 73% saline. Compound 25 solution was administered intragastrically to SD rats at a dosage of 5.0 mg/kg. Blood samples were taken at the following time points: 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours after administration.

To prepare the solution, test compound 25 was dissolved in a mixture of 5% DMSO + 20% (30% solyutol) + 2% 1M NaOH + 73% saline. Compound 25 solution was administered to SD rats at a dosage of 1 mg/kg via intravenous bolus injection. Blood sampling was performed at the following time points: 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours after administration.

To prepare the solution, test compounds 84 and 96 were dissolved in a mixture of 5% DMSO + 10% PEG400 + 85% (28% HP-β-CD) saline. A solution of compounds 84 and 96 was administered intragastrically to SD rats at a dosage of 5.0 mg/kg. Blood samples were taken at the following time points: 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours after administration.

To prepare the solution, test compounds 84 and 96 were dissolved in a mixture of 5% DMSO + 10% PEG400 + 85% (28% HP-β-CD) saline. A solution of compounds 84 and 96 was administered to SD rats at a dosage of 1 mg/kg via intravenous bolus injection. Blood sampling was performed at the following time points: 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours after administration.

The animals were fixed. The tail of the animal was heated in a water bath for 10 minutes before each sampling moment. Approximately 100 μl of blood was taken from the tail vein, which after collection was placed in an anticoagulation tube containing EDTA-K2. Blood samples were centrifuged at 8000 rpm at 4° C. for 6 minutes to obtain plasma samples. Plasma was prepared within 30 minutes after blood sampling and stored in a freezer at -80°C until testing.

Sample analysis method

The test plasma sample was removed from the freezer, which was kept at -80°C, left to warm at room temperature and vortexed for 5 minutes. Twenty (20) µl of the plasma sample was carefully pipetted into a 1.5 ml centrifuge tube, after which 200 µl of the working solution of the internal standard (tolbutamide in methanol) at a concentration of 100 ng/ml was added, mixed, vortexed for 5 minutes and then centrifuged at 12,000 rpm for 5 minutes. 50 μl of the liquid above the pellet was taken and carefully placed with a pipette into 96-well plates, in which water was previously placed in an amount of 150 μl/well; vortexed for 5 minutes and analyzed by LC-MS/MS. The injection volume of compounds 25, 65, 76, 84, 90, 96, 107, 154 and 158 was 20, 20, 5, 10 µl, respectively.

Data processing method

The concentration of the test samples was determined using the Analyst 1.6.1 software supplied by AB SCIEX. Microsoft Excel was used to calculate the mean, standard deviation, coefficient of variation, and other parameters (directly output by Analyst 1.6.1 and not calculated). Pharmacokinetic parameters were calculated using Pharsight Phoenix 6.1 NCA software (T _max is the mean).

The results are presented in Table 7:

As can be seen from the data above in the Table, the compounds according to the invention have very good pharmacokinetic characteristics.

Experimental Example 5

Method for assessing the enzymatic properties of PDE

Test substance: Compounds according to the invention prepared according to the respective embodiments of the invention

1. Experimental materials and equipment

Enzyme PDE1A1 (BPS, catalog no.: 60010)

PDE3A enzyme (BPS, catalog no.: 60030)

Enzyme PDE5A1 (BPS, catalog no.: 60050)

Enzyme PDE8A1 (BPS, catalog no.: 60080)

384-well plate (Perkin Elmer, catalog no.: 6007279)

2. Procedure of the experiment

Preparation of Compounds: Compounds were prepared for long-term storage as a 10 mM stock solution of compounds in DMSO.

The resulting compound stock solution was diluted 100 times with DMSO to give a working compound stock solution at a concentration of 100 μM, and then the working compound stock solution was diluted 3 times with DMSO to give 10 concentration gradients of the diluted compound stock solution (100x).

Incubation with Compounds: An Echo pipetting system was used to pipette a diluted stock solution of a compound into a 384-well plate to pipette very small amounts of liquid. To each compound well was added 200 panoliters of diluted compound stock solution and 10 μl of PDE1A1, PDE3A, PDE5A1 and PDE8A1 enzyme solution. After centrifugation at 1000 rpm for 1 minute, the mixture was incubated for 15 minutes at room temperature. Then 10 µl of the substrate mixture was added. After centrifugation at 1000 rpm for 1 minute, the mixture was incubated with shock for 30 minutes at room temperature. Finally, stop solution was added to stop the reaction in the system. The mixture was incubated with shock for 60 minutes at room temperature. At maximum reading (Max), the compound was displaced by solvent. At the minimum reading (Min), both compound and enzyme solution were replaced with solvent.

Measurements: Fluorescence values (F) at 480 nm/535 nm were determined using a microplate reader.

Calculation: The rate of inhibition was calculated according to the following formula, and the IC ₅₀ values were approximated using GraphPad Prism 5.0 software:

3. Test results are presented in Table 8:

Experimental Conclusion: The compounds of the present invention have no significant inhibitory effect on the PDE1A1, PDE3A, PDE5A15 or PDE8A1 enzymes and have a relatively high selectivity for PDE9.

It should be noted that, unless otherwise indicated, each of the Experimental Examples discussed above may be implemented in a conventional manner known in the art, and a third party may also be involved to perform each of the Experimental Examples; for example, Experimental Examples 1 and 5 discussed above are provided by a third party.

The above are only preferred embodiments of the present invention, and the present invention is not limited to them. Any modifications, equivalents, improvements and similar changes consistent with the subject and scope of the present invention should be included in the scope of the present invention.

Claims (106)

1. The compound represented by formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof,

Where each of X ₁ , X ₂ , X ₃ and X ₄ independently represents CR ₃ or N, and X ₁ X ₂ , X ₃ and X ₄ do not simultaneously represent CR ₃ ; R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, carboxyl, cyano, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, piperazinyl and morpholinyl, C _{1- 6} alkylcarbonyl, aminocarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, 4-6 membered heterocyclylcarbonyl selected from piperazinylcarbonyl and azetidinylcarbonyl, and pyrimidinyloxy, where C _1-6 alkyl, C _1-6 alkoxy, (C _1-6 alkyl) ₂ amino, C _2-8 alkynyl, 4-6 membered saturated nitrogen containing heterocyclyl selected from azetidinyl, piperazinyl and morpholinyl, aminocarbonyl, C _1-6 alkylaminocarbonyl and 4-6 membered heterocyclylcarbonyl selected from piperazinylcarbonyl and azetidinylcarbonyl are unsubstituted or optionally substituted with one or two groups independently selected from the group consisting of hydroxy, amino, cyano, C _1-6 alkyl, C _1-6 alkoxy, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonyloxy, C _3-6 cycloalkyl, halogenated C _1-6 alkyl and 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl which is unsubstituted or optionally substituted with C _1-6 alkyl; L is a bond, -NH-(CH ₂ ) _t -, and t is 0, 1, 2 or 3; ring A is selected from the group consisting of a 5-7 membered saturated monoheterocyclyl having an N atom as a heteroatom, a 7-11 membered saturated spiroheterocyclyl having 1 or 2 heteroatoms selected from O and N, and an aryl having from 6 to 14 carbon atoms; each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C _1-6 alkyl optionally substituted with hydroxy, C _1-6 alkoxy, and a 5-membered heteroaryl containing 1 to 3 heteroatoms selected from S and N; m is 0.1, 2 or 3; And R ₂ is hydrogen; provided that: when X ₁ is N and X ₂ , X ₃ and X ₄ are each independently CR ₃ , A is not phenyl; when X ₁ and X ₃ are each independently N and X ₂ and X ₄ are each independently CR ₃ , A is not phenyl; And when X ₃ is N and X ₁ , X ₂ and X ₄ are each independently CR ₃ , A is not phenyl. 2. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 1, Where each of X ₁ , X ₂ , X ₃ and X ₄ independently represents CR ₃ or N, and X ₁ , X ₂ , X ₃ and X ₄ do not simultaneously represent CR ₃ ; R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, carboxyl, cyano, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered saturated nitrogen heterocyclyl selected from azetidinyl, piperazinyl and morpholinyl, C _{1- 6} alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl, where C 1-6 alkyl _, C _1-6 alkoxy, (C _1-6 alkyl) ₂ amino, C _2-8 alkynyl, 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, piperazinyl and morpholinyl, C _1-6 alkylaminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or two groups independently selected from the group consisting of hydroxy, amino, cyano, C _{1- 6} alkyl, C _1-6 alkoxy, (C _1-6 alkyl) ₂ amino, C _3-6 cycloalkyl, C _1-6 alkylcarbonyloxy and unsubstituted or C _1-6 alkyl substituted 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, pyrrolidinide and piperidinyl; L is a bond, -NH-(CH ₂ ) _t , and t is 0, 1, 2 or 3; ring A is selected from the group consisting of a 5-7 membered saturated monoheterocyclyl having an N atom as a heteroatom, a 7-11 membered saturated spiroheterocyclyl having 1 or 2 heteroatoms selected from O and N, and an aryl having from 6 to 14 carbon atoms; each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C _1-6 alkyl optionally substituted with hydroxy, C _1-6 alkoxy, and a 5-membered heteroaryl containing 1 to 3 heteroatoms selected from S and N; m is 0.1, 2 or 3; And R ₂ is hydrogen, provided that: when X ₁ is N and X ₂ , X ₃ and X ₄ are each independently CR ₃ , A is not phenyl; when X ₁ and X ₃ are each independently N and X ₂ and X ₄ are each independently CR ₃ , A is not phenyl; And when X ₃ is N and X ₁ , X ₂ and X ₄ are each independently CR ₃ , A is not phenyl. 3. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 2, wherein the compound has the structure represented by formula (II):

Where X ₂ , X ₃ , X ₄ , R ₁ , R ₂ , R ₃ , ring A, L and m are as defined in paragraph 2, and X ₂ , X ₃ and X ₄ do not simultaneously represent CR ₃ ; provided that when X ₃ is N and X ₂ and X ₄ are each independently CR ₃ , A is not phenyl. 4. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 3, Where each of X ₂ , X ₃ and X ₄ independently represents CR ₃ or N, and X ₂ , X ₃ and X ₄ do not simultaneously represent CR ₃ ; R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, carboxyl, cyano, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 alkylamino, (C _1-6 alkyl) ₂ amino, C _2-8 alkenyl, C _2-8 alkynyl, C _1-6 alkylsulfonyl, C _1-6 alkylthio, C _3-6 cycloalkyl, 4-6 membered saturated nitrogen heterocyclyl selected from azetidinyl, piperazinyl and morpholinyl, C _{1- 6} alkylcarbonyl, C _1-6 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl and aminocarbonyl, where C 1-6 alkyl _{, C 1-6 alkoxy, (C 1-6 alkyl ) 2} amino, C _2-8 alkynyl, 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, piperazinyl and morpholinyl, C _1-6 alkylaminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or two groups independently selected from the group consisting of hydroxy, amino, cyano, C _{1- 6} alkyl, C _1-6 alkoxy, (C _1-6 alkyl) ₂ amino, C _1-6 alkylcarbonyloxy, C _3-6 cycloalkyl and unsubstituted or substituted C _1-6 alkyl 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl; L is a bond; ring A is a 5-7 membered saturated monoheterocyclyl having an N atom as a heteroatom, or a 7-11 membered saturated spiroheterocyclyl having 1 or 2 heteroatoms selected from O and N; each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C _1-6 alkyl optionally substituted with hydroxy, C _1-6 alkoxy, and a 5-membered heteroaryl having 1 to 3 heteroatoms selected from S and N; m is 0, 1 or 2; And R ₂ is hydrogen. 5. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 4, Where X ₂ is N and each of X ₃ and X ₄ is independently CR ₃ or N; R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, cyano, halogen, carboxyl, C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, C _2-6 alkenyl, C _2-6 alkynyl, C _1-4 alkylcarbonyl, C _1-4 alkylaminocarbonyl, (C _1-6 alkyl) ₂ aminocarbonyl, C _1-4 alkylsulfonyl, C _1-4 alkylthio, aminocarbonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl, where C _1-4 alkyl, C _1-4 alkoxy, (C _1-4 alkyl) ₂ amino, C _2-6 alkynyl, C _1-4 alkylaminocarbonyl, aminocarbonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl are unsubstituted or optionally substituted with one or two groups independently selected from the group consisting of hydroxy, amino, cyano, C _1-4 alkyl, C _1-4 alkoxy, (C _1-4 alkyl) ₂ amino, cyclopropyl, C _{1- 4} alkylcarbonyloxy and an unsubstituted or C _1-4 alkyl substituted 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl; L is a bond; ring A is a 5-7-membered saturated monoheterocyclyl having an N atom as a heteroatom, or a 7-11-membered saturated spiro-heterocyclyl having 1 or 2 heteroatoms selected from O and N, wherein the 7-11-membered saturated spiro-heterocyclyl contains at least one N, and ring A is connected to L through an N atom; each R ₁ is independently selected from the group consisting of hydrogen, hydroxy, cyano, halogen, C _1-4 alkyl optionally substituted with hydroxy, C _1-4 alkoxy, pyrazolyl, thiazolyl, and triazolyl; And m is 0, 1, or 2. 6. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 4 or 5, Where X ₂ is N, X ₃ is CR ₃ and X ₄ is CR ₃ or N; R ₃ at each occurrence is independently selected from the group consisting of hydrogen, halogen, C _1-4 alkyl, C _1-4 alkoxy, morpholinyl, C _2-6 alkenyl, C _1-4 alkylaminocarbonyl, (C _1-4 alkyl) ₂ aminocarbonyl and aminocarbonyl, wherein C _1-4 alkyl, C _1-4 alkoxy, morpholinyl, C _1-4 alkylaminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or two groups independently selected from the group consisting of hydroxy, C _1-4 alkoxy , cyclopropyl, amino (C _1-4 alkyl) ₂ amino and unsubstituted or substituted C _1-4 alkyl 4-6 membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl; L is a bond; ring A is a 5-7 membered saturated monoheterocyclyl having an N atom as a heteroatom, ring A is connected to L through an N atom; each R ₁ is independently selected from the group consisting of hydrogen, halogen, C _1-4 alkyl optionally substituted with hydroxy, C _1-4 alkoxy, pyrazolyl, thiazolyl and triazolyl; And m is 0, 1, or 2. 7. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 6, wherein ring A is selected from the group consisting of

8. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 6, Where X ₂ is N and X ₃ and X ₄ are each independently CR ₃ ; R ₃ at each occurrence is independently selected from the group consisting of hydrogen, halogen, C _1-4 alkyl, C _1-4 alkoxy, C _2-6 alkenyl, C _1-4 alkylaminocarbonyl and aminocarbonyl, where C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkylaminocarbonyl and aminocarbonyl are unsubstituted or optionally substituted with one or two groups independently selected from the group consisting of hydroxy, amino, C _1-4 alkyl, C _1-4 alkoxy, cyclopropyl, (C _{1 -4} alkyl) ₂ amino and unsubstituted or substituted C _1-4 alkyl 4-6-membered saturated nitrogen-containing heterocyclyl selected from azetidinyl, pyrrolidinyl and piperidinyl; L is a bond; ring A is

each R ₁ is independently selected from the group consisting of hydrogen, C _1-4 alkyl optionally substituted with hydroxy, and C _1-4 alkoxy; And m is 0, 1, or 2. 9. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 6, Where X ₂ is N and X ₃ and X ₄ are each independently CR ₃ ; R ₃ at each occurrence is independently selected from the group consisting of hydrogen, halogen, C _1-4 alkyl and morpholinyl, where C _1-4 alkyl is unsubstituted or substituted with one or two hydroxy groups; L is a bond; Ring A is

And each R ₁ is independently selected from the group consisting of pyrazolyl, thiazolyl and triazolyl. 10. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 4 or 5, Where R ₃ at each occurrence is independently selected from the group consisting of hydrogen, amino, cyano, halogen, carboxyl, C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkylcarbonyl, C _2-6 alkynyl, C _1-4 alkylaminocarbonyl, (C _1-4 alkyl) ₂ aminocarbonyl, C _1-4 alkylthio, C _1-4 alkylsulfonyl, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, azetidinyl, morpholinyl, piperazinyl, C _2-6 alkenyl and cyclopropyl, where C _1-4 alkyl, C _1-4 alkoxy, C _2-6 alkynyl, C _1-4 alkylaminocarbonyl, (C _1-4 alkyl) ₂ amino, azetidinyl, morpholinyl, piperazinyl and cyclopropyl are unsubstituted or optional substituted with one or two groups independently selected from the group consisting of hydroxy, amino, C _1-4 alkyl, (C _1-4 alkyl) ₂ amino, cyclopropyl, and C _1-4 alkylcarbonyloxy; L is a bond; ring A is a 7-11 membered saturated spiroheterocyclyl having 1 or 2 heteroatoms selected from O and N, where the 7-11 membered saturated spiroheterocyclyl contains at least one N and ring A is connected to L through an N atom; each R ₁ is independently selected from the group consisting of hydrogen, cyano, halogen, hydroxy, and unsubstituted or hydroxy-substituted C _1-4 alkyl; And m is 0, 1, or 2. 11. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 10, wherein ring A is selected from the group consisting of

12. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 10, Where X ₂ is N and X ₃ and X ₄ are each independently CR ₃ or N; R ₃ at each occurrence is independently selected from the group consisting of hydrogen, cyano, amino, halogen, carboxyl, C _1-4 alkyl, C _1-4 alkoxy, C _2-6 alkenyl, C _1-4 alkylcarbonyl, C _2-6 alkynyl, C _1-4 alkylamino, (C _1-4 alkyl) ₂ amino, C _1-4 alkylaminocarbonyl, C _1-4 alkylthio, C _1-4 alkylsulfonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl, where C _1-4 alkyl , C _1-4 alkoxy, C _2-6 alkynyl, (C _1-4 alkyl) ₂ amino, C _1-4 alkylaminocarbonyl, cyclopropyl, azetidinyl, morpholinyl and piperazinyl are unsubstituted or optionally substituted with one or two groups independently selected from the group , consisting of hydroxy, amino, C _1-4 alkyl, (C _1-4 alkyl) ₂ amino, cyclopropyl and C _1-4 alkylcarbonyloxy; L is a bond; ring A is selected from the group consisting of

And m is 0. 13. A compound or a pharmaceutically acceptable salt or stereoisomer thereof according to claim 4 or 5, Where L is a bond; X ₂ is N and each of X ₃ and X ₄ is independently CR ₃ or N; And Ring A is selected from the group consisting of:

14. The compound or its pharmaceutically acceptable salt or stereoisomer according to any one of paragraphs. 1-4, where the compound is selected from the group consisting of:

15. A pharmaceutical composition that inhibits PDE9, comprising a therapeutically effective amount of a compound or a pharmaceutically acceptable salt or stereoisomer thereof according to any one of paragraphs. 1-14 and a pharmaceutically acceptable carrier. 16. The use of a compound or a pharmaceutically acceptable salt or stereoisomer thereof according to any one of paragraphs. 1-14 or a pharmaceutical composition according to claim 15 for the preparation of a medicament for the treatment or prevention of a disease mediated by PDE9, 17. Use according to claim 16, wherein the PDE9 mediated disease is a cognitive disorder caused by a disorder of the central nervous system. 18. Use according to claim 17, wherein the cognitive disorder is selected from the group consisting of impaired perception, attention, memory, and learning ability.