WO2025087291A1 - Irak4 targeted degradation agent compound, and use thereof - Google Patents

Abstract

The present disclosure relates to a IRAK4 degradation agent compound, and specifically provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof. The compound has the substituents and structural features described in the present application. The application also discloses a pharmaceutical composition comprising the compound represented by formula (I) or a pharmaceutically acceptable salt of the composition, and a pharmaceutical use of the compound or the pharmaceutically acceptable salt thereof.

Description

IRAK4 targeted degradation agent compounds and applications thereof

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the rights and priority of the following two Chinese invention patent applications, the entire contents of which are hereby incorporated by reference into this document in their entirety: Patent Application No. 202311383103.6 filed with the State Intellectual Property Office on October 23, 2023, and Patent Application No. 202410167580.7 filed with the State Intellectual Property Office on February 5, 2024.

Technical Field

The present disclosure relates to compounds or pharmaceutically acceptable salts thereof as IRAK4 degraders, methods for preparing the same, pharmaceutical compositions containing the same, and uses of the same or pharmaceutically acceptable salts thereof or the pharmaceutical compositions in preventing or treating diseases or conditions mediated by IRAK4.

Background Art

Interleukin-1 receptor kinase 4 (IRAK4) is a serine/threonine protein kinase, a core regulator of innate immune response, and plays an important role in activating the immune system. IL-1 receptors and Toll-like receptors are activated after binding to their ligands, and then recruit intracellular myeloid differentiation factor MyD88. MyD88 further recruits IRAK4 through its N-terminal death domain. After autophosphorylation, IRAK4 recruits and activates IRAK1 and IRAK2 to form a MyD88-IRAK4-IRAK1/2 complex, which transmits signals downstream to the E3 ubiquitin ligase TNF receptor-associated factor (TRAF6), activates the serine/threonine kinase TAK1, and further activates the NF-κB and MAPK signaling pathways, causing the release of a variety of inflammatory cytokines and proliferation-related factors. Studies have shown that overactivation of IRAK4 is associated with a variety of autoimmune diseases, such as atopic dermatitis, suppurative hidradenitis, and rheumatoid arthritis. However, IRAK4 deficiency is not lethal to individuals, and in adulthood, the risk of bacterial infection is reduced, which is no different from that of healthy people.

In addition, the scaffold function of IRAK4 can also regulate downstream signaling pathways, and is independent of its kinase function. Studies have shown that mouse macrophages with IRAK4 kinase function loss can still be induced to activate the NF-κB signaling pathway; IRAK4 kinase inhibitors cannot block the IRAK4-mediated NF-κB signaling pathway in human monocytes, suggesting that this may be related to the insufficient efficacy of IRAK4 kinase inhibitors under clinical development.

Proteolysis Targeting Chimeria (PROTAC) is a bifunctional molecule, one end of which is a small molecule inhibitor that recognizes the target protein through a linker, and the other end is an E3 ubiquitin ligase ligand that can recognize E3 ubiquitin ligase, thereby forming a ternary complex. After the target protein is ubiquitinated, it is degraded in vivo through the ubiquitin-proteasome pathway. Traditional IRAK4 small molecule inhibitors can only block the kinase activity of IRAK4, while IRAK4 PROTAC can degrade intracellular IRAK4, thereby simultaneously blocking the biological activity of IRAK4 kinase activity and scaffold function, and more effectively inhibiting the release of downstream inflammatory factors mediated by IRAK4. Therefore, it is necessary to develop new IRAK4 PROTAC drugs for the treatment of diseases or conditions related to IRAK4.

Summary of the invention

The present disclosure relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof:

TL-Linker-DIM

(I)

in:

The DIM is a ligand compound capable of binding to E3 ubiquitin ligase;

The Linker is a linking group that covalently binds at least one TL and at least one DIM;

The TL is a group as shown below:

in:

A ¹ , A ² , A ³ , A ⁴ are independently selected from CH, CR ¹ or N; R ¹ is selected from deuterium, halogen, CN, -C(O)NR ^1a R ^1b , NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₀ aryl, 5-10 membered heteroaryl or 4-14 membered heterocyclyl, wherein the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₀ aryl, 5-10 membered heteroaryl or 4-14 membered heterocyclyl is optionally substituted with one or more ^Ra ;

R ^1a is selected from H, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl, wherein the C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl is optionally substituted by one or more R ^1c ;

R ^1b is selected from H or C ₁ -C ₆ alkyl;

One of B ¹ and B ² is selected from C-NHR ² , C-OR ² or C-CHR ^2a R ^2b , and the other is selected from CH or N;

R ² , R ^2a , R ^2b are independently selected from H, deuterium, halogen, CN, NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-8 membered heterocyclyl, phenyl or 5-6 membered heteroaryl, wherein the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-8 membered heterocyclyl, phenyl or 5-6 membered heteroaryl is optionally substituted by one or more R ^b ;

Ring Q is a 5-10 membered heteroaromatic ring, and the 5-10 membered heteroaromatic ring is optionally substituted by one or more R ^c ;

One of B ³ and B ⁴ is C and is connected to ring Q, and the other is selected from CH or N;

X is selected from S, O or NR ³ ;

R ³ is selected from H, deuterium, halogen, CN, NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, phenyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl, wherein the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, phenyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl is optionally substituted with one or more R ^d ;

^Ra , ^Ric , ^Rb , ^Rc , ^Rd are independently selected from deuterium, halogen, OH, CN, _NH2 , =O, _C1 - _C6 alkyl, _C3 - _C6 cycloalkyl, 5-6-membered heteroaryl or 4-8-membered heterocyclyl, wherein the OH, _NH2 , _C1 - _C6 alkyl, C3- _C6 cycloalkyl, 5-6-membered heteroaryl or _4-8 -membered heterocyclyl is optionally substituted by one or more ^Re ;

R ^e is selected from deuterium, halogen, OH, NH ₂ , ═O, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, CN, COOH, C(O)(C ₁ -C ₃ alkyl), CONH ₂ , C(O)O(C ₁ -C ₃ alkyl), S(O) ₂ (C ₁ -C ₃ alkyl), C ₃ -C ₆ cycloalkyl or 4-6 membered heterocyclyl.

In some embodiments, one of B ¹ and B ² is C-NHR ² , and the other is selected from CH or N.

In some embodiments, B ¹ is C-NHR ² and B ² is selected from CH or N.

In some embodiments, B ¹ is C-NHR ² and B ² is N.

In some embodiments, ^B3 is C and is attached to ring Q, and ^B4 is selected from CH or N.

In some embodiments, ^B3 is C and is attached to ring Q, and ^B4 is CH.

In some embodiments, ^B3 is selected from CH or N, and ^B4 is C and is attached to ring Q.

In some embodiments, B ¹ is C-NHR ² , B ² is selected from CH or N, B ³ is C and is attached to ring Q, and B ⁴ is selected from CH or N.

In some embodiments, R ² , R ^2a , R ^2b are independently selected from H, deuterium, OH, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-7 membered heterocyclyl, phenyl or 5-6 membered heteroaryl, and the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-7 membered heterocyclyl, phenyl or 5-6 membered heteroaryl are optionally substituted with one or more R ^b .

In some embodiments, B ¹ is C-NHR ² , R ² is selected from H, deuterium, OH, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-7 membered heterocyclyl, phenyl or 5-6 membered heteroaryl, and the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-7 membered heterocyclyl, phenyl or 5-6 membered heteroaryl is optionally substituted with one or more R ^b .

In some embodiments, B ¹ is C-NHR ² , R ² is selected from H, deuterium, OH, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, or 4-7 membered heterocyclyl, wherein the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, or 4-7 membered heterocyclyl is optionally substituted with one or more R ^b .

In some embodiments, B ¹ is C-NHR ² , R ² is selected from H, deuterium, OH or C ₁ -C ₆ alkyl, said OH, NH ₂ or C ₁ -C ₆ alkyl being optionally substituted with one or more R ^b .

In some embodiments, B ¹ is C-NHR ² , and R ² is selected from 4-7 membered heterocyclyl or C ₁ -C ₄ alkyl, wherein the 4-7 membered heterocyclyl or C ₁ -C ₄ alkyl is optionally substituted with R ^b .

In some embodiments, B ¹ is C-NHR ² , and R ² is selected from tetrahydropyranyl, oxetanyl, or C ₁ -C ₄ alkyl optionally substituted with R ^b .

In some embodiments, B ¹ is C-NHR ² , and R ² is selected from tetrahydropyranyl, oxetanyl, -CH ₂ CN, or isopropyl.

In some embodiments, B ¹ is C-NHR ² , R ² is selected from Or _-CH2CN .

In some embodiments, B ¹ is C-NHR ² , and R ² is selected from C ₁ -C ₄ alkyl, such as isopropyl.

In some embodiments, A ¹ and A ⁴ are selected from CH, CR ¹ or N, and A ² and A ³ are selected from CH or CR ¹ .

In some embodiments, A ⁴ is selected from CH, CR ¹ or N, and A ¹ , A ² , A ³ are selected from CH or CR ¹ .

In some embodiments, A ⁴ is selected from CH or N, A ² is CR ¹ , and A ¹ and A ³ are both CH.

In some embodiments, A ⁴ is selected from CH, A ² is CR ¹ , and A ¹ and A ³ are both CH.

In some embodiments, A ¹ is selected from CH, CR ¹ or N, and A ² , A ³ , A ⁴ are selected from CH or CR ¹ .

In some embodiments, A ¹ is selected from CH or N, A ³ is CR ¹ , and A ² and A ⁴ are both CH.

In some embodiments, A ¹ is selected from CH, A ³ is CR ¹ , and A ² and A ⁴ are both CH.

In some embodiments, R ¹ is selected from deuterium, halogen, CN, -C(O)NR ^1a R ^1b , NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, or 4-7 membered heterocyclyl, wherein the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, or 4-7 membered heterocyclyl is optionally substituted with one or more ^Ra .

In some embodiments, R ¹ is selected from deuterium, halogen, CN, NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, or 4-7 membered heterocyclyl, wherein the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, or 4-7 membered heterocyclyl is optionally substituted with one or more ^Ra .

In some embodiments, R ¹ is selected from deuterium, halogen, CN, OH, or C ₁ -C ₄ alkyl, wherein the OH or C ₁ -C ₄ alkyl is optionally substituted with one or more ^Ra .

In some embodiments, R ¹ is selected from CN.

In some embodiments, R ^1a is selected from H or C _{1 -C 6} alkyl, which is optionally substituted with one or more R ^1c ; _R ^1b is selected from H or C ₁ -C ₆ alkyl.

In some embodiments, R ^1a is selected from C ₃ -C ₆ cycloalkyl, 5-6 membered heteroaryl or 4-7 membered heterocyclyl, wherein the C ₃ -C ₆ cycloalkyl, 5-6 membered heteroaryl or 4-7 membered heterocyclyl is optionally substituted with one or more R ^1c ; R ^1b is selected from H or C ₁ -C ₆ alkyl.

In some embodiments, Ring Q is a 5-6 membered heteroaryl ring optionally substituted with one or more R ^c .

In some embodiments, Ring Q is a 5-membered heteroaromatic ring optionally substituted with one or more R ^c , such as the following groups optionally substituted with one or more R ^c : pyrazolyl, imidazolyl, triazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl or thiadiazolyl.

In some embodiments, Ring Q is the following groups optionally substituted with one or more R ^c : thiadiazolyl, triazolyl, pyrazolyl.

In some embodiments, Ring Q is the following groups optionally substituted with one or more R ^c : thiadiazolyl, triazolyl.

In some embodiments, Ring Q is

In some embodiments, Ring Q is

In some embodiments, R ³ is selected from H, deuterium, halogen, CN, NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, said OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl optionally substituted with one or more R ^d .

In some embodiments, R ³ _is selected from H, deuterium, C ₁ -C ₆ alkyl _, or C _{3 -C 6} cycloalkyl _, which is optionally substituted with one or more R ^d .

In some embodiments, R ³ is selected from H, deuterium, or C ₁ -C ₄ alkyl optionally substituted with one or more R ^d .

In some embodiments, R ³ is selected from H or C ₁ -C ₄ alkyl.

In some embodiments, X is selected from S, O, or NH.

In some embodiments, X is selected from NR ³ .

In some embodiments, X is selected from NH.

In some embodiments, ^Ra , ^R1c , ^Rb , ^Rc , ^Rd are independently selected from deuterium, halogen, OH, CN, _NH2 , =O, _C1 - _C6 alkyl, _C3 - _C6 cycloalkyl, or 4-7 membered heterocyclyl, and the OH, _NH2 , _C1 - _C6 alkyl, _C3 - _C6 cycloalkyl, or 4-7 membered heterocyclyl is optionally substituted with one or more ^Re .

In some embodiments, ^Ra , ^Ric , ^Rb , ^Rc , ^Rd are independently selected from deuterium, halogen, OH, CN, _NH2 , =O, _C1 - _C6 alkyl, or 4-8 membered heterocyclyl, wherein said OH, _NH2 , _C1 - _C6 alkyl, or 4-8 membered heterocyclyl is optionally substituted with one or more ^Re .

In some embodiments, ^Ra , ^R1c , ^Rb , ^Rc , ^Rd are independently selected from deuterium, halogen, OH, CN, _NH2 , =0 or _C1 - _C6 alkyl, wherein said OH, _NH2 or _C1 - _C6 alkyl is optionally substituted with one or more ^Re .

In some embodiments, R ^b is CN.

In some embodiments, ^Re is selected from deuterium, halogen, OH, _NH2 , _C1 - _C3 alkyl, _C1 - _C3 alkoxy, COOH, C(O)( _C1 - _C3 alkyl), _CONH2 , C(O)O( _C1 - _C3 alkyl), _C3 - _C6 cycloalkyl, or 4-6 membered heterocyclyl.

In some embodiments, ^Re is selected from halogen, _C1 - _C3 alkyl, _C1 - _C3 alkoxy, COOH, C(O)( _C1 - _C3 alkyl), _CONH2 , or C(O)O( _C1 - _C3 alkyl).

In some embodiments, the TL is selected from the structure shown below (TL-1):

wherein B ² and B ⁴ are independently selected from CH or N, and A ¹ , A ² , A ³ , A ⁴ , X, R ² and Q are as defined above.

In some embodiments, the TL is selected from the structure shown below (TL-2):

wherein: B ² and B ⁴ are independently selected from CH or N, A ⁴ is selected from CH or N, n is selected from 0, 1 or 2, and R ¹ , R ² , X and ring Q are as defined above.

In some embodiments, the TL is further selected from the group shown below:

In some embodiments, the TL is further selected from the group shown below:

In some embodiments, the TL is selected from the following structures:

In some embodiments, the TL is

In some embodiments, the Linker is a linking group that covalently binds a TL and a DIM.

In some embodiments, the linker is selected from: -L ^A -, -L ^B -, -R ^1L -, -R ^2L -, -Q ¹ -, -Q ² -,

wherein: ^-LA- , ^-LB- are independently selected from a chemical bond, -O-, -S-, -NR 3'-, -CR ^4'R5'- , -CR 4'R5' ^- NR ^3'- , -CR ^{4'R5' -} O-, -C(O ⁾ -, -CR ^{4'R5' -} C(O)-, -S(O)-, -S(O) ^2- , -C(S ⁾ -, -C(O)O- or -C(O)NR ^6'- _;

R ^1L and R ^2L are independently selected from a chemical bond, -C(O)-, alkylene, heteroalkylene, alkenylene and alkynylene, wherein the alkylene, heteroalkylene, alkenylene and alkynylene are optionally substituted by a group selected from the group consisting of halogen, alkyl, alkoxy, haloalkyl, OH, hydroxyalkyl, CN, NH ₂ , =O, cycloalkyl, heterocyclyl, aryl, heteroaryl;

Q ¹ , Q ² , Q ³ and Q ⁴ are independently selected from cycloalkylene, heterocyclylene, arylene, heteroarylene or cycloalkenylene, wherein the cycloalkylene, heterocyclylene, arylene, heteroarylene and cycloalkenylene are each independently optionally substituted by a group selected from the following: halogen, alkyl, alkoxy, haloalkyl, OH, hydroxyalkyl, CN, NH ₂ , =O, cycloalkyl, heterocyclyl, aryl, heteroaryl;

R ^3′ is selected from H, alkyl, heteroalkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

R ^4′ and R ^5′ are each independently selected from H, halogen, alkyl, alkoxy, haloalkyl, OH, hydroxyalkyl, CN, NH ₂ , ═O, cycloalkyl, heterocyclyl, aryl or heteroaryl;

R ^6′ is selected from H, alkyl, heteroalkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl.

In some embodiments, the Linker is In some embodiments, the Linker is

In some embodiments, R ^1L and R ^2L are each independently selected from a bond, _-C (O)-, or C _{1 -C 3} alkylene, which is optionally substituted with C ₁ -C ₃ alkyl.

In some embodiments, Q ¹ , Q ² , Q ³ and Q ⁴ are independently selected from C ₃ -C ₈ cycloalkylene or 5-10 membered heterocyclylene, wherein the C ₃ -C ₈ cycloalkylene or 5-10 membered heterocyclylene is optionally substituted by a group selected from halogen, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, C ₁ -C ₃ haloalkyl, OH, C ₁ -C ₃ hydroxyalkyl or NH ₂ .

In some embodiments, the linker is selected from the following structures: bond, -O-, -C(O)-, -CH ₂ -,

In some embodiments, the linker is selected from: a bond, -O-, -C(O)-, -CH ₂ -,

In some embodiments, the Linker is selected from

In some embodiments, the Linker is selected from

In some embodiments, the Linker is selected from

In some embodiments, the DIM is selected from ligand compounds capable of binding to a cereblon-type E3 ubiquitin ligase.

In some embodiments, the DIM is selected from the structure shown in formula (DIM-1) or (DIM-2):

in:

Selected from

Y is a chemical bond, or Y is selected from _YA , O, NH, NR _E , C(O)O, C(O)NR _E ', NR _E'C (O), YA- _NH , _YA -NR _E , _{YA- C} (O), YA- _C (O)O, YA- _OC (O), YA-C(O)NR _E ' or YA- _{NR E'C} (O), wherein _YA is selected from _C1 - _C6 alkylene, _C2 - _C6 alkenylene or _C2 - _C6 alkynylene;

X' is selected from C(O) or C( _RA ) ₂ ; XA _{- XB} is selected from C( _RA )=N or C( _RA ) ₂ -C( _RA ) ₂ ;

Each _RA is independently selected from H or C ₁ -C ₃ alkyl, wherein the C ₁ -C ₃ alkyl is optionally substituted with C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl;

Each _RA ' is independently selected from _C1 - _C3 alkyl;

Each _RB is independently selected from H or _C1 - _C3 alkyl, or two _RBs together with the atoms to which they are attached form C(O), _C3 - _C6 cycloalkyl, _C3 - _C6 cycloalkenyl or 4-6 membered heterocyclyl;

R _C is selected from H, halogen or C ₁ -C ₃ alkyl;

Each R _D is independently selected from halogen, NO ₂ , NH ₂ , OH, COOH, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy;

Each _RE is independently selected from C1- _C6 alkyl, _C2 - _C6 alkenyl, _C3 - _C8 cycloalkyl, 3-8 membered heterocycloalkyl, C(O)-C1- _C6 alkyl _, C(O) _-C2 - _C6 alkenyl, C(O) _{-C3 -C8 cycloalkyl} or C(O)-3-8 membered heterocycloalkyl, and the _RE is optionally substituted by a group selected from the following: halogen halogen, N(R _a ) ₂ , NHC(O)R _a , NHC(O)OR _a , OR _b , C ₃ -C ₈ cycloalkyl, 3-8 membered heterocycloalkyl, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl, wherein the C ₃ -C ₈ cycloalkyl, 3-8 membered heterocycloalkyl, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl is optionally further substituted by a group selected from halogen, NH ₂ , CN, NO ₂ , OH, COOH, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy;

R _E 'is selected from H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₃ -C ₈ cycloalkyl or 3-8 membered heterocycloalkyl, wherein the C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₃ -C ₈ cycloalkyl or 3-8 membered heterocycloalkyl is optionally substituted by a group selected from the group consisting of halogen, N(R _a ) ₂ , NHC(O)R _a , NHC(O)OR _a , OR _b , C ₃ -C ₈ cycloalkyl, 3-8 membered heterocycloalkyl, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl, wherein the C ₃ -C _{8 cycloalkyl, 3-8 membered heterocycloalkyl, C 6 -C 10 aryl or 5-10 membered heteroaryl is optionally further substituted by a group selected from the group consisting of halogen, NH 2 , CN, NO 2 , OH, COOH, C 1 -C 6 alkyl, C 1 -C 6 alkenyl, C 3 -C 8 cycloalkyl or 3-8 membered} heterocycloalkyl C ₁ -C ₆ haloalkyl, C 1 -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy;

Each _Ra is independently selected from H or _C1 - _C6 alkyl;

R _b is selected from H or p-toluenesulfonyl;

t is selected from 0 or 1;

m is selected from 0, 1, 2 or 3;

p is selected from 0, 1 or 2.

In some embodiments, the DIM is further selected from the structure shown in formula (DIM-3) or (DIM-4):

wherein the ring A, Y, _RA , _RA ', _RB , _RC , _RD , m and p are as defined above.

In some embodiments, the DIM is further selected from the structures shown in formula (DIM-5), (DIM-6), (DIM-7) or (DIM-8):

wherein Y, X', _XA - _XB , _RA , _RA ', _RB , _RC , _RD , m and p are as defined above.

In some embodiments, the DIM is further selected from the structure shown in formula (DIM-9) or (DIM-10):

wherein Y, X', _XA - _XB , _RA , _RA ', _RB , _RC , _RD , m and p are as defined above.

In some embodiments, the DIM is selected from the structure shown in formula (DIM-11):

in:

_XC is selected from a chemical bond, _-CH2- , -CHCF3-, _-SO2- , _-S (O)-, -P(O)R'-, -P(O)(OR')-, -P(O)( _NR'2 )-, -C(O)-, -C(S)- or

_XD is selected from C, N or Si;

X _E is selected from a chemical bond, -CR' ₂ -, -NR'-, -O-, -S- or -SiR' ₂ -;

_RF is absent or is selected from H, _deuterium , halogen, CN, -OR', -SR', -S(O)R', -S(O) _2R ', -NR'2, -P(O)(OR') ₂ , -P(O)( _NR'2 )OR', -P(O)( _NR'2 ) ₂ , -Si(OH) _2R ', -Si(OH) _R'2 , _-SiR'3 , or _C1 - _C4 alkyl _;

Each _RG is independently selected from H, deuterium, _RH , halogen, CN, -NO2, _-OR ', -SR', _-NR'2 , _-SiR'3 , -S(O) _2R ', -S(O) _2NR'2 , -S(O)R _' , -C(O)R', -C(O)OR', -C(O) _NR'2 , -C(O)N(R')OR', -C(R') _2N (R')C(O)R', -C(R') _2N (R')C(O) _NR'2 , -OC(O)R', -OC(O) _NR'2 , -OP(O)R'2, -OP(O)(OR') ₂ , _-OP (O ₎ (OR')NR'2, -OP(O)( _NR'2 ) ₂ , -N(R')C(O)OR', -N(R')C(O)R', -N(R')C(O)NR' ₂ , -N(R')S(O) ₂ R' , -N(R')P(O)R' ₂ , -N(R')P(O)(OR') ₂ , -N(R')P(O)(OR')NR' ₂ or -N(R')P(O)(NR' ₂ ) ₂ ;

Each _RH is independently selected from _C1 - _C6 alkyl, phenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl;

Ring E, Ring F, Ring G are independently selected from phenyl, 5-6 membered heteroaryl, C ₅ -C ₇ cycloalkyl, C ₅ -C ₇ cycloalkenyl or 5-7 membered heterocyclyl, wherein Ring E, Ring F and Ring G are each optionally further substituted by =O;

L ¹ is selected from a chemical bond, a C ₁ -C ₃ alkylene group, a C ₂ -C ₃ alkenylene group or a C ₂ -C ₃ alkynylene group, wherein any _{one or} two methylene groups in the C ₁ -C ₃ alkylene group, the C ₂ -C ₃ alkenylene group or the C 2 -C 3 alkynylene group are optionally replaced by the following groups: -O-, -C(O)-, -C(S)-, -C(R') ₂ -, -CH(R')-, -C(F) ₂ -, -N(R')-, -S- or -S(O) ₂ -;

Each R' is independently selected from H, C ₁ -C ₆ alkyl, phenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl, or two R' and the atoms to which they are attached together form a 4-7 membered heterocyclyl or 5-6 membered heteroaryl;

q is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.

In some embodiments, the DIM is selected from the structure shown in formula (DIM-11'):

wherein X _C , _RF , _RG , q, ring E, ring F and ring G are as defined in formula (DIM-11).

In some embodiments, the DIM is selected from the structure shown in formula (DIM-12):

wherein: ring H is selected from C ₅ -C ₉ cycloalkyl, C ₅ -C ₉ cycloalkenyl or 5-9 membered heterocyclyl, said C ₅ -C ₉ cycloalkyl, C ₅ -C ₉ cycloalkenyl or 5-9 membered heterocyclyl is optionally substituted by =O; k is selected from 0, 1, 2, 3 or 4; X _C , X _D , X _E , _RF , _RG , L ¹ and ring E are as defined in formula (DIM-11).

In some embodiments, the DIM is selected from the structure shown in formula (DIM-12'):

wherein X _C , _RF , _RG , k, ring E and ring H are as defined in formula (DIM-12).

In some embodiments, the DIM is selected from the structure shown in formula (DIM-13):

wherein X _C , X _D , X _E , _RF , _RG , L ¹ , ring E and k are as defined in formula (DIM-12).

In some embodiments, the DIM is selected from the structure shown in (DIM-13'):

wherein X _C , _RF , _RG , ring E and k are as defined in formula (DIM-13).

In some embodiments, the DIM is selected from the structure shown in formula (DIM-1), formula (DIM-3), formula (DIM-5), formula (DIM-12), formula (DIM-12'), formula (DIM-13) or formula (DIM-13').

In some embodiments, the DIM is selected from the structures shown in formula (DIM-3), formula (DIM-5), formula (DIM-12), formula (DIM-12'), formula (DIM-13) or formula (DIM-13').

In some embodiments, the DIM is selected from the following structures:

In some embodiments, the DIM is selected from the following structures:

In some embodiments, the DIM is selected from the following structures:

In some embodiments, the DIM is selected from the following structures:

In some embodiments, the compound of formula (I) of the present disclosure or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

On the other hand, the present disclosure provides a pharmaceutical composition comprising a compound or a specific compound represented by any of the above general formulae of the present disclosure or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

On the other hand, the present disclosure provides a method for treating a disease mediated by IRAK4 in a mammal, comprising administering a therapeutically effective amount of a compound represented by any of the above general formulas of the present disclosure or a specific compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof to a mammal, preferably a human, in need of such treatment.

On the other hand, the present disclosure provides a method for treating tumors, inflammatory diseases, neurodegenerative diseases or autoimmune diseases in mammals, comprising administering a therapeutically effective amount of a compound represented by any of the above general formulas or a specific compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof to a mammal, preferably a human, in need of such treatment.

On the other hand, the present disclosure provides use of a compound represented by any of the above general formulas or a specific compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating a disease mediated by IRAK4.

On the other hand, the present disclosure provides the use of a compound represented by any of the above general formulas or a specific compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a medicament for preventing or treating tumors, inflammatory diseases, neurodegenerative diseases or autoimmune diseases.

On the other hand, the present disclosure provides the use of a compound represented by any of the above general formulas or a specific compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in preventing or treating a disease mediated by IRAK4.

On the other hand, the present disclosure provides the use of the compound represented by any of the above general formulas or specific compounds or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof in preventing or treating tumors, inflammatory diseases, neurodegenerative diseases or autoimmune diseases.

In another aspect, the present disclosure provides a compound of any of the above general formulas or specific compounds or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for preventing or treating a disease mediated by IRAK4.

In another aspect, the present disclosure provides any of the above general formula compounds or specific compounds or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, for preventing or treating tumors, inflammatory diseases, neurodegenerative diseases or autoimmune diseases.

In some embodiments, the disease mediated by IRAK4 is selected from a tumor, an inflammatory disease, a neurodegenerative disease, or an autoimmune disease.

Any embodiment of any aspect of the present disclosure can be combined with other embodiments without contradiction. In addition, in any embodiment of any aspect of the present disclosure, any technical feature can be applied to the technical feature in other embodiments without contradiction.

Definitions and explanations of terms

Unless otherwise specified, the terms used in this disclosure have the following meanings. The groups and term definitions recorded in this disclosure, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be arbitrarily combined and combined with each other. A specific term should not be considered uncertain or unclear in the absence of a special definition, but should be understood according to the common meaning in the art. When a trade name appears in this article, it is intended to refer to its corresponding The product or its active ingredients.

In this article Indicates the connection site.

The term "capable of binding" means capable of measurably binding to a target (eg, a ligand of an E3 ubiquitin ligase is capable of forming a covalent bond with a cysteine of an E3 ubiquitin ligase, etc.).

The term "ubiquitin ligase" refers to a family of proteins that facilitate the transfer of ubiquitin to a specific substrate protein to target the substrate protein for degradation. E3 ubiquitin ligases, alone or in complex with E2 ubiquitin ligases, are responsible for transferring ubiquitin to target proteins. Typically, ubiquitin ligases participate in polyubiquitination, such that a second ubiquitin is attached to a first ubiquitin; a third ubiquitin is attached to a second ubiquitin, and so on. Polyubiquitination marks proteins for degradation by the proteasome. However, there are some ubiquitination events that are limited to monoubiquitination, in which ubiquitin ligases only add a single ubiquitin to a substrate molecule. Monoubiquitinated proteins are not targeted to the proteasome for degradation, but can change their cellular location or function, for example, by binding to other proteins with domains capable of binding ubiquitin. To complicate matters further, E3 ubiquitin ligases can target different lysines on ubiquitin to make chains.

The term "target protein" refers to proteins and peptides having any biological function or activity, including structural, regulatory, hormonal, enzymatic, genetic, immune, contractile, storage, transport, and signal transduction. In some embodiments, a target protein refers to a protein or polypeptide that binds to a compound of the present disclosure and can be degraded.

The term "tautomer" refers to functional group isomers resulting from the rapid movement of an atom in two positions in a molecule. Compounds of the present disclosure may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Tautomers generally exist in equilibrium, and attempts to separate a single tautomer usually result in a mixture whose physical and chemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical characteristics within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; while in phenols, the enol form predominates. The present disclosure encompasses all tautomeric forms of the compounds.

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

The compounds of the present disclosure may have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms or asymmetric double bonds, so the compounds of the present disclosure may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis and trans isomers, E-type and Z-type geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures or other mixtures thereof, such as mixtures enriched in enantiomers or diastereomers, all of which are within the definition of the compounds of the present disclosure and their mixtures. Additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms or asymmetric phosphorus atoms may be present in substituents such as alkyl, and all of these isomers and their mixtures involved in all substituents are also within the definition of the compounds of the present disclosure. Compounds of the present disclosure containing an asymmetric atom can be isolated in optically pure or racemic forms, which can be resolved from racemic mixtures or synthesized by using chiral starting materials or chiral reagents.

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are replaced, and oxo will not occur on an aromatic group.

The term "optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes both the occurrence and non-occurrence of the event or circumstance. For example, an ethyl group is "optionally" substituted with a halogen, which means that the ethyl group may be unsubstituted (CH ₂ CH ₃ ), monosubstituted (CH ₂ CH ₂ F, CH ₂ CH ₂ Cl, etc.), polysubstituted (CHFCH ₂ F, CH ₂ CHF ₂ , CHFCH ₂ Cl, CH ₂ CHCl ₂ , etc.) or fully substituted (CF ₂ CF ₃ , CF ₂ CCl ₃ , CCl ₂ CCl ₃ , etc.). It will be understood by those skilled in the art that for any group containing one or more substituents, no substitution or substitution pattern that is sterically impossible to exist and/or cannot be synthesized will be introduced.

The term "optionally substituted" means that the compound may be substituted or unsubstituted, and unless otherwise specified, the type and amount of the substituents may be any on the basis of what is chemically feasible.

The term "replaced" means that a specific atom or group can be replaced by another specified atom or group. For example, CH ₂ in -CH ₂ CH ₂ CH ₂ - can be replaced by O, S or NH to obtain -CH ₂ OCH ₂ -, -OCH ₂ CH ₂ -, -CH ₂ SCH ₂ -, -SCH ₂ CH ₂ -, -CH ₂ NHCH ₂ - or -NHCH ₂ CH ₂ -.

When any variable (e.g., ^Ra , ^Rb ) occurs more than once in the composition or structure of a compound, its definition in each case is independent. For example, if a group is substituted by 2 ^Rb , each ^Rb has independent options; for the group N( _C1 - _C6 alkyl) ₂ , when _C1 - _C6 alkyl is substituted by ^Rb , the two _C1 - _C6 alkyls have independent ^Rb options.

When the number of a linking group is 0, such as -(CH ₂ ) ₀ -, it means that the linking group is a bond.

When one of the variables is selected from a chemical bond or does not exist, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a bond, it means that the structure is actually AZ.

When the linking group mentioned in this article does not specify its connection direction, its connection direction is arbitrary. When L ¹ is selected from "C ₁ -C ₃ alkylene-O", L ¹ can connect ring Q and R ¹ from left to right to form "ring QC ₁ -C ₃ alkylene-OR ¹ ", or connect ring Q and R ¹ from right to left to form "ring QOC ₁ -C ₃ alkylene-R ¹ ".

_Cm - _Cn herein refers to an integer number of carbon atoms in the range of mn or m to n. For example, " _C1 - _C10 " means that the group may have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms or 10 carbon atoms. Similarly, m-membered to n-membered means that the number of ring atoms is m to n, for example, 5-14-membered ring includes 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered ring, 10-membered ring, 11-membered ring, 12-membered ring, 13-membered ring and 14-membered ring, and also includes any range from n to m, for example, 5-14-membered ring includes 6-14-membered ring, 6-11-membered ring, 5-10-membered ring, 6-10-membered ring, 6-8-membered ring, etc.

The term "alkyl" refers to a hydrocarbon group of the general formula _{CnH2n +1} , which is a straight or branched group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, and more preferably an alkyl group containing 1 to 6 carbon atoms. The term " _C1 - _C10 alkyl" is understood to mean a straight or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Specific examples of the alkyl group include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc.; the term "C ₁ - ... The term "C 1 -C ₄ alkyl" may be understood to mean an alkyl group having 1 to 6 carbon atoms, specific examples of which include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, and the like. The term "C ₁ -C ₄ alkyl" may be understood to mean a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, or 4 carbon atoms. The term "C ₁ -C ₃ alkyl" may be understood to mean a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, or 3 carbon atoms. The “C ₁ -C ₁₀ alkyl” may include “C ₁ -C ₆ alkyl”, “C ₁ -C ₄ alkyl” or “C ₁ -C ₃ alkyl”, and the “C ₁ -C ₆ alkyl” may further include “C ₁ -C ₄ alkyl” or “C ₁ -C ₃ alkyl”, and the “C ₁ -C ₄ alkyl” may further include “C ₁ -C ₃ alkyl”.

The term "heteroalkyl" refers to an alkyl group in which one or more _-CH2- are replaced by a heteroatom selected from NH, O and S, or one or more -CH- are replaced by N; wherein the alkyl group is as defined above.

The term "haloalkyl" refers to a group obtained by further replacing the alkyl with halogen, such as "C ₁ -C ₆ haloalkyl" refers to a C ₁ -C ₆ alkyl further replaced with halogen. The term "hydroxyalkyl" refers to a group obtained by further replacing the alkyl with OH.

The term "alkylene" refers to a saturated straight or branched aliphatic hydrocarbon group having two residues derived from the same carbon atom or two different carbon atoms of an alkane radical by removing two hydrogen atoms, and is a straight or branched group containing 1 to 20 carbon atoms, preferably an alkylene group containing 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12) carbon atoms, and more preferably an alkylene group containing 1 to 6 carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene, -CH(CH ₃ )-, -CH ₂ CH ₂ -, -CH(CH ₂ CH ₃ )-, -CH ₂ CH(CH ₃ )-, -CH ₂ CH ₂ CH ₂ -, -CH ₂ CH ₂ CH _{2 -} , and the like. The term "C ₁ -C ₆ alkylene group" is understood to mean an alkylene group having 1 to 6 carbon atoms. The term "C ₁ -C ₃ alkylene" is understood to mean an alkylene group having 1 to 3 carbon atoms. Preferably, "C ₁ -C ₆ alkylene" may include "C ₁ -C ₃ alkylene". The term "heteroalkylene" refers to an alkylene group in which one or more -CH ₂ - groups are substituted by heteroatoms selected from N, O and S; wherein the alkylene group is as defined above.

The term "alkoxy" refers to a monovalent group generated by the loss of a hydrogen atom from a hydroxyl group of a straight or branched alcohol, and can be understood as "alkyloxy" or "alkyl-O-", wherein the definition of alkyl is as described above. The term "C ₁ -C ₁₀ alkoxy" can be understood as "C ₁ -C ₁₀ alkyloxy" or "C _{1 -C 10} alkyl-O-"; the term "C ₁ -C ₆ alkoxy" can be understood as "C 1 -C ₆ alkyloxy" or "C ₁ -C ₆ alkyl-O-". The "C ₁ -C ₁₀ alkoxy" can include "C ₁ -C ₆ alkoxy" and "C ₁ -C ₃ alkoxy" and the like, and the "C ₁ -C ₆ alkoxy" can further include "C ₁ -C ₃ alkoxy".

The term "haloalkoxy" refers to a group obtained by further substituted with halogen by the alkoxy group. For example, "C ₁ -C ₆ haloalkoxy" refers to a C ₁ -C ₆ alkoxy group further substituted with halogen.

The term "alkenyl" refers to an unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms, which is straight or branched and has 2 to 20 carbon atoms and has at least one double bond. The term "C ₂ -C ₁₀ alkenyl" is understood to mean a straight or branched unsaturated monovalent hydrocarbon group, which contains one or more double bonds and has 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, and the term "C ₂ -C ₆ alkenyl" is understood to mean a straight or branched unsaturated monovalent hydrocarbon group, which contains one or more double bonds and has 2, 3, 4, 5 or 6 carbon atoms. "C ₂ -C ₁₀ alkenyl" is preferably "C ₂ -C ₆ alkenyl" or "C ₂ -C ₄ alkenyl", "C ₂ -C ₆ alkenyl" is further preferably "C ₂ -C ₄ alkenyl", and further preferably C ₂ or C ₃ alkenyl. It should be understood that in the case where the alkenyl group contains more than one double bond, the double bonds may be separated or conjugated from each other. Specific examples of the alkenyl group include, but are not limited to, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl or (Z)-1-methylprop-1-enyl, etc.

The term "alkenylene" refers to a residue derived from the same carbon atom or two different carbon atoms of a parent alkene by removing two hydrogen atoms, wherein alkenyl is as defined above. The term "C ₂ -C ₆ alkenylene" is understood as an alkenylene having 2 to 6 carbon atoms. The term "C ₂ -C ₃ alkenylene" is understood as an alkenylene having 2 or 3 carbon atoms. Preferably, "C ₂ -C ₆ alkenylene" includes "C ₂ -C ₃ alkenylene".

The term "alkynyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms, containing 2 to 20 carbon atoms and having at least one triple bond. The term "C ₂ -C ₁₀ alkynyl" may be understood to mean a linear or branched unsaturated monovalent hydrocarbon group containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. The term "C ₂ -C ₆ alkynyl" may be understood to mean a linear or branched unsaturated monovalent hydrocarbon group containing one or more triple bonds and having 2, 3, 4, 5 or 6 carbon atoms. Examples of "C ₂ -C ₆ alkynyl" include, but are not limited to, ethynyl (-C≡CH ₃ , -CH 2 C≡CH 3 , -CH ₂ C≡CH ), but-1-ynyl, but-2-ynyl or but-3-ynyl. “C ₂ -C ₁₀ alkynyl” may include “C ₂ -C ₆ alkynyl” or “C ₂ -C ₃ alkynyl”, and “C ₂ -C ₆ alkynyl” may include “C ₂ -C ₃ alkynyl”. Examples of “C ₂ -C ₃ alkynyl” include ethynyl (—C≡CH), prop-1-ynyl (—C≡CCH ₃ ) or prop-2-ynyl (propargyl).

The term "alkynylene" refers to a residue derived from the same carbon atom or two different carbon atoms of a parent alkyne by removing two hydrogen atoms, wherein the definition of alkynyl is as shown above. The term "C ₂ -C ₆ alkynylene" is to be understood as an alkynylene having 2 to 6 carbon atoms. The term "C ₂ -C ₃ alkynylene" is to be understood as an alkynylene having 2 or 3 carbon atoms. Preferably, "C ₂ -C ₆ alkynylene" includes "C ₂ -C ₃ alkynylene".

The term "cycloalkyl" refers to a fully saturated carbocyclic ring that exists in the form of a monocyclic, cyclic, bridged or spirocyclic ring. Unless otherwise indicated, the carbocyclic ring is generally a 3- to 10-membered ring. The term "C ₃ -C ₁₀ cycloalkyl" is understood to mean a saturated monovalent monocyclic, cyclic, spirocyclic or bridged ring having 3 to 10 carbon atoms. The term "C ₃ -C ₈ cycloalkyl" is understood to mean a saturated monovalent monocyclic, cyclic, spirocyclic or bridged ring having 3 to 8 carbon atoms. The term "C ₃ -C ₆ cycloalkyl" is understood to mean a saturated monovalent monocyclic, cyclic, spirocyclic or bridged ring having 3 to 6 carbon atoms, and specific examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. The term "C ₅ -C ₉ cycloalkyl" is understood to mean a saturated monovalent monocyclic, cyclic, spirocyclic or bridged ring having 5 to 9 carbon atoms. The term " _C5 - _C7 cycloalkyl" is understood to mean a saturated monovalent monocyclic, cyclopentyl, spiro or bridged ring having 5 to 7 carbon atoms. Specific examples of the cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decyl, and the like. The term " _C3 - _C10 cycloalkyl" may include " _C3 - _C8 cycloalkyl", " _C3 - _C6 cycloalkyl", " _C5 - _C9 cycloalkyl" or " _C5 - _C7 cycloalkyl", the term " _C3 - _C8 cycloalkyl" may include " _C3 - _C6 cycloalkyl" or " _C5 - _C7 cycloalkyl", the term " _C5 - _C9 cycloalkyl" may include " _C5 - _C7 cycloalkyl".

The term "cycloalkyloxy" may be understood as "cycloalkyl-O-". For example, "C ₃ -C ₆ cycloalkyloxy" may be understood as "C ₃ -C ₆ cycloalkyl-O-".

The term "cycloalkenyl" refers to a non-aromatic carbocyclic ring that is not fully saturated and exists in the form of a monocyclic ring, a cyclic ring, a bridged ring or a spirocyclic ring. Unless otherwise indicated, the carbocyclic ring is generally a 3-10 membered ring. Specific examples of the cycloalkenyl include, but are not limited to, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl or cycloheptadienyl. The term "C ₅ -C ₁₀ cycloalkenyl" refers to a non-aromatic carbocyclic ring that is not fully saturated and exists in the form of a monocyclic ring, a cyclic ring, a bridged ring or a spirocyclic ring, and has 5-10 carbon atoms. The term "C ₅ -C ₉ cycloalkenyl" refers to a non-aromatic carbocyclic ring that is not fully saturated and exists in the form of a monocyclic ring, a cyclic ring, a bridged ring or a spirocyclic ring, and has 5, 6, 7, 8 or 9 carbon atoms. The term "C ₅ -C ₇ cycloalkenyl" refers to a non-aromatic carbocyclic ring that is not fully saturated and exists in the form of a monocyclic ring, a cyclic ring, a bridged ring or a spirocyclic ring, etc., and has 5, 6 or 7 carbon atoms. The term "C ₃ -C ₆ cycloalkenyl" refers to a non-aromatic carbocyclic ring that is not fully saturated and exists in the form of a monocyclic ring, a cyclic ring, a bridged ring or a spirocyclic ring, etc., and has 3, 4, 5 or 6 carbon atoms. The term "C ₅ -C ₁₀ cycloalkenyl" may include "C ₅ -C ₉ cycloalkenyl" or "C ₅ -C ₇ cycloalkenyl", and the term "C ₅ -C ₉ cycloalkenyl" may include "C ₅ -C ₇ cycloalkenyl".

The term "heterocyclyl" refers to a fully saturated or partially saturated (not a heteroaromatic group having aromaticity as a whole) monovalent monocyclic, fused, spiro or bridged ring group, which contains 1, 2, 3, 4 or 5 heteroatoms or heteroatom groups (i.e., an atom group containing heteroatoms) in the ring atoms, wherein the "heteroatoms or heteroatom groups" include, but are not limited to, nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(=O) _2- , -S(=O)- and optionally substituted -NH-, -S(=O)(=NH)-, -C(=O)NH-, -C(=NH)-, -S(=O) _2NH- , S(=O)NH- or -NHC(=O)NH-, etc., and usually contains 3 to 20 ring atoms. The term "4-14 membered heterocyclyl" refers to a heterocyclyl having 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, and the ring atoms thereof contain 1 to 5 heteroatoms or heteroatom groups independently selected from the above. "4-14 membered heterocyclyl" may include "6-14 membered heterocyclyl", "6-11 membered heterocyclyl", "6-10 membered heterocyclyl", "6-8 membered heterocyclyl", "4-10 membered heterocyclyl", "4-8 membered heterocyclyl", "4-7 membered heterocyclyl", "4-6 membered heterocyclyl", "5-10 membered heterocyclyl", "5-9 membered heterocyclyl", "5-8 membered heterocyclyl" or "5-7 membered heterocyclyl". The term "5-10 membered heterocyclyl" may include "5-9 membered heterocyclyl", "5-8 membered heterocyclyl", "5-7 membered heterocyclyl", "6-10 membered heterocyclyl" or "6-8 membered heterocyclyl". The term "4-10 membered heterocyclyl" refers to a heterocyclyl having 4, 5, 6, 7, 8, 9 or 10 ring atoms, and the ring atoms thereof contain 1-5 heteroatoms or heteroatom groups independently selected from the above. "4-10 membered heterocyclyl" includes "4-8 membered heterocyclyl", "4-7 membered heterocyclyl" or "4-6 membered heterocyclyl", wherein specific examples of 4 membered heterocyclyl include but are not limited to azetidinyl or oxetanyl; specific examples of 5 membered heterocyclyl include but are not limited to tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl or 2,5-dihydro-1H-pyrrolyl; Specific examples of 6-membered heterocyclic groups include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridinyl or 4H-[1,3,4]thiadiazinyl; specific examples of 7-membered heterocyclic groups include, but are not limited to, diazepanyl. The heterocyclic group may also be a bicyclic group, wherein specific examples of 5,5-membered bicyclic groups include, but are not limited to, hexahydrocyclopenta[c]pyrrole-2(1H)-yl; specific examples of 5,6-membered bicyclic groups include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazine-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group may be a benzo-fused ring group of the above-mentioned 4-7-membered heterocyclic group, specific examples of which include, but are not limited to, dihydroisoquinolinyl and the like. The “4-10 membered heterocyclyl” may include the ranges of “5-10 membered heterocyclyl”, “5-9 membered heterocyclyl”, “5-8 membered heterocyclyl”, “5-7 membered heterocyclyl”, “5-6 membered heterocyclyl”, “6-10 membered heterocyclyl”, “6-8 membered heterocyclyl”, “4-8 membered heterocyclyl”, “4-7 membered heterocyclyl”, “4-6 membered heterocyclyl”, “4-10 membered heterocycloalkyl”, “5-10 membered heterocycloalkyl”, “4-7 membered heterocycloalkyl”, “5-6 membered heterocycloalkyl”, “6-8 membered heterocycloalkyl”, and the like, and the “4-7 membered heterocyclyl” may further include the ranges of “4-6 membered heterocyclyl”, “5-7 membered heterocyclyl”, “5-6 membered heterocyclyl”, “4-7 membered heterocyclyl”, “4-6 membered heterocycloalkyl”, “5-7 membered heterocycloalkyl”, “5-6 membered heterocycloalkyl”, and the like. Although some bicyclic heterocyclic groups in the present disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclic group as a whole is still non-aromatic.

The term "heterocyclyloxy" may be understood as "heterocyclyl-O-".

The term "heterocycloalkyl" refers to a fully saturated monovalent cyclic group in the form of a monocyclic, fused, bridged or spirocyclic ring, wherein the ring atoms of the ring contain 1, 2, 3, 4 or 5 heteroatoms or heteroatomic groups (i.e., an atomic group containing heteroatoms). The "heteroatoms or heteroatomic groups" include, but are not limited to, nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(＝O) ₂ -, -S(＝O)- and optionally substituted -NH-, -S(＝O)(＝NH)-, -C(＝O)NH-, -C(＝NH)-, -S(＝O) ₂ NH-, S(＝O)NH- or -NHC(＝O)NH-, etc., which usually contain 3 to 20 ring atoms. The term "3-10 membered heterocycloalkyl" refers to a heterocycloalkyl group having 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms, and containing 1 to 5 heteroatoms or heteroatom groups independently selected from the above-mentioned heteroatoms. “3-10 membered heterocycloalkyl” includes “3-8 membered heterocycloalkyl”, wherein specific examples of 4 membered heterocycloalkyl include but are not limited to azetidinyl, oxetanyl or thietanyl; specific examples of 5 membered heterocycloalkyl include but are not limited to tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, thiazolidinyl, imidazolidinyl or tetrahydropyrazolyl; specific examples of 6 membered heterocycloalkyl include but are not limited to piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, 1,4-thioxanyl, 1,4-dioxane, thiomorpholinyl, 1,3-dithianyl or 1,4-dithianyl; specific examples of 7 membered heterocycloalkyl include but are not limited to azepanyl, oxetanyl or thiepanyl.

The term "heterocycloalkyloxy" may be read as "heterocycloalkyl-O-".

The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group with a conjugated π electron system. The aryl group may have 6-20 carbon atoms, 6-14 carbon atoms or 6-12 carbon atoms. The term "C ₆ -C ₂₀ aryl" is understood to mean a monovalent aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 20 carbon atoms. In particular, it is a ring having 6 carbon atoms ("C ₆ aryl"), such as phenyl; or a ring having 9 carbon atoms ("C ₉ aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C ₁₀ aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl; or a ring having 13 carbon atoms ("C ₁₃ aryl"), such as fluorenyl; or a ring having 14 carbon atoms ("C ₁₄ aryl"), such as anthracenyl. The term "C ₆ -C ₁₀ aryl" is understood to mean a monovalent aromatic all-carbon monocyclic or bicyclic group having 6 to 10 carbon atoms, in particular a ring having 6 carbon atoms ("C ₆ aryl"), such as phenyl; or a ring having 9 carbon atoms ("C ₉ aryl"), such as indenyl; or a ring having 10 carbon atoms ("C ₁₀ aryl"), such as naphthyl.

The term "aryloxy" may be understood as "aryl-O-".

The term "heteroaryl" refers to a monocyclic or fused polycyclic system with aromaticity, which contains at least one, preferably 1, 2, 3 or 4 ring atoms selected from N, O, S, and the remaining ring atoms are carbon 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14-membered aromatic ring radical. Heteroaryl is preferably a 5-10-membered, more preferably a 5- or 6-membered heteroaryl. The term "5-10-membered heteroaryl" is understood to include such monovalent monocyclic or bicyclic aromatic ring systems: it has 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms, and it contains 1-5, preferably 1-3 heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl, and the like, and benzo derivatives thereof, such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl, and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl or isoquinolyl, and the like; or azinyl, indolizinyl, purinyl, and the like, and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, and the like. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and containing 1-3, preferably 1-2 heteroatoms independently selected from N, O and S. The term "6 membered heteroaryl" refers to an aromatic ring system having 6 ring atoms, and containing 1-3, preferably 1-2 heteroatoms independently selected from N, O and S. The term "5-10 membered heteroaryl" may include "5-6 membered heteroaryl" or "6 membered heteroaryl", and the term "5-6 membered heteroaryl" may include "6 membered heteroaryl".

The term "halo" or "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "hydroxy" refers to an -OH group.

The term "cyano" refers to a -CN group.

The term "amino" refers to a _-NH2 group.

The term "nitro" refers to the _-NO2 group.

The term "therapeutically effective amount" means an amount of a compound of the present disclosure that (i) treats or prevents a particular disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of a compound of the present disclosure that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one skilled in the art based on their own knowledge and the present disclosure.

The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to a salt of a pharmaceutically acceptable acid or base, including a salt formed between a compound and an inorganic acid or an organic acid, and a salt formed between a compound and an inorganic base or an organic base.

The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure or their salts and pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present disclosure to an organism.

The term "pharmaceutically acceptable excipients" refers to those excipients that have no significant irritation to the organism and do not impair the biological activity and performance of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

The word "comprise" or "comprises" and its English variations such as comprises or comprising should be construed in an open and non-exclusive sense, ie, "including but not limited to".

The present disclosure also includes isotopically labeled compounds of the present disclosure that are identical to those described herein, but in which one or more atoms are replaced by atoms having an atomic mass or mass number different from that normally found in nature. Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ¹²³ I, ¹²⁵ I, and ³⁶ Cl, etc., respectively.

Certain isotopically labeled compounds of the disclosure (e.g., labeled with ³ H and ¹⁴ C) are useful in compound and/or substrate tissue distribution assays. Tritiated (i.e., ³ H) and carbon-14 (i.e., ¹⁴ C) isotopes are particularly preferred for their ease of preparation and detectability. Positron emitting isotopes, such as ¹⁵ O, ¹³ N, ¹¹ C, and ¹⁸ F, can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the disclosure can generally be prepared by the following procedures, similar to those disclosed in the schemes and/or examples below, by substituting an isotopically labeled reagent for an unlabeled reagent.

The pharmaceutical compositions of the present disclosure can be prepared by combining the compounds of the present disclosure with suitable pharmaceutically acceptable excipients, for example, they can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.

Typical routes of administration of the compounds of the present disclosure, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

The pharmaceutical composition of the present disclosure can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, emulsification methods, freeze-drying methods, and the like.

In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present disclosure to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, etc., for oral administration to patients.

Solid oral compositions can be prepared by conventional mixing, filling or tableting methods. For example, they can be obtained by mixing the active compound with a solid excipient, optionally grinding the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into particles to obtain a tablet or dragee core. Suitable excipients include, but are not limited to, adhesives, diluents, disintegrants, lubricants, glidants or flavoring agents, etc.

The pharmaceutical composition may also be suitable for parenteral administration, such as sterile solutions, suspensions or lyophilized products in appropriate unit dosage forms.

The dosage of the compound or composition used in the treatment methods of the present disclosure will generally vary with the severity of the disease, the patient's body weight and the relative efficacy of the compound, however, as a general guide, a suitable daily dosage of the compound of formula (I) described herein is 0.01 mg/kg to 1000 mg/kg.

The compounds of the present disclosure can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present disclosure.

DETAILED DESCRIPTION

The present disclosure is described in detail below by specific embodiments, but it is not intended to limit the present disclosure in any way. Various specific embodiments of the present disclosure have been described in detail herein, and it will be apparent to those skilled in the art that various changes and modifications may be made to the specific embodiments of the present disclosure without departing from the spirit and scope of the present disclosure.

The chemical reactions of the specific embodiments of the present disclosure are performed in a suitable solvent, which must be suitable for the chemical changes of the present disclosure and the reagents and materials required. In order to obtain the compounds of the present disclosure, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes based on the existing embodiments.

All reagents used in this disclosure were commercially available and used without further purification.

Unless otherwise specified, the ratios expressed for mixed solvents are volume mixing ratios. Unless otherwise specified, % means wt%.

The structure of the compound is determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS). The unit of NMR shift is 10 ^-6 (ppm). The solvents for NMR determination are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, D ₂ O, CF ₃ COOD, etc., and the internal standard is tetramethylsilane (TMS); "IC ₅₀ " refers to the half inhibitory concentration, which refers to the concentration when half of the maximum inhibitory effect is achieved.

Explanation of terms or abbreviations:

t-Bu: tert-butyl; t-BuOK: potassium tert-butoxide; t-BuNO ₂ : tert-butyl nitrite; TEA: triethylamine; THF: tetrahydrofuran; Me: methyl; MeOH: methanol; MeI: methyl iodide; EtOH: ethanol; ⁱ PrNH ₂ : isopropylamine; DCM: dichloromethane; DCE: dichloroethane; DMF: N,N-dimethylformamide; DIEA: N,N-diisopropylethylamine; NMP: N-methylpyrrolidone; HATU: O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate; Boc: tert-butyloxycarbonyl; Bn: benzyl; NaBH(OAc) ₃ : sodium triacetoxyborohydride; AcOH: acetic acid; EA/EtOAc: ethyl acetate; TFA: trifluoroacetic acid; Pd(PPh ₃ ) ₄ : tetrakis(triphenylphosphine)palladium; Pd(PPh ₃ ) ₂ Cl ₂ : bis(triphenylphosphine)palladium dichloride; Pd(dppf)Cl ₂ : 1,1-bis(diphenylphosphino)ferrocenepalladium chloride; Pd ₂ (dba) ₃ : tris(dibenzylideneacetone)dipalladium; Pd(OAc) ₂ : palladium acetate; NaOAc: sodium acetate; Py: pyridine; Ms: methanesulfonyl; MsCl: methanesulfonyl chloride; MsOH: methanesulfonic acid; Tf ₂ O: trifluoromethanesulfonic anhydride; dioxane: 1,4-dioxane; toluene/PhMe: toluene; acetone: acetone; DMSO: dimethyl sulfoxide; STAB: sodium triacetoxyborohydride; m-CPBA: m-chloroperbenzoic acid; DMK: acetone; TCFH: N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate; M: mole/liter; TBS: tert-butyldimethylsilyl; TBSCl: tert-butylchlorodimethylsilane; IMZ: imidazole; DMP: Dess-Martin periodinane; MPa: megapascal; Bn: benzyl; PMB: p-methoxybenzyl; DMP: Dess-Martin periodinane; RuPhos Pd G3: methanesulfonate (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl) palladium(II); RuPhos: 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl; JohnPhos: 2-(di-tert-butylphosphino)biphenyl; XantPhos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene; TAA: tert-amyl alcohol; MW: microwave; Cu(Tc): cuprous thiophene-2-carboxylate; BPin: 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl; Pd-PEPPSI-Ipent-Cl: dichloro[1,3-bis(2 ,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II); JohnPhos: 2-(di-tert-butylphosphino)biphenyl; NMI: 1-methylimidazole; PivOH: trimethylacetic acid; Xphos-Pd-G3: methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II); NIS: N-iodosuccinimide; ACN: acetonitrile; IBX: 2-iodoacylbenzoic acid; NADPH: reduced nicotinamide adenine dinucleotide phosphate; HEPES: 4-hydroxyethylpiperazineethanesulfonic acid; NMDG: N-methyl-D-glucamine.

The eluent mentioned below may be a mixed eluent formed by two or more solvents, the ratio of which is the volume ratio of each solvent.

Intermediate A: Synthesis of 4-(isopropylamino)-3-(5-(piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-5H-pyrido[3,2-b]indole-7-carbonitrile

Step 1: Synthesis of 5-bromo-2-chloro-N-isopropyl-3-nitropyridin-4-amine (A-2)

5-Bromo-2,4-dichloro-3-nitropyridine (A-1, 4.00 g, 14.71 mmol) was dissolved in N,N-dimethylformamide (20 mL), and triethylamine (2.98 g, 29.42 mmol) and isopropylamine (869.64 mg, 14.71 mmol) were added to react at room temperature for 3 h. LC-MS detection showed that the reaction was complete. The reaction solution was diluted with ethyl acetate (50 mL), then washed with saturated brine (20 mL × 3), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 0-30% gradient elution) to obtain the title compound (3.44 g).

MS m/z(ESI):294.0,296.0[M+H] ⁺ .

Step 2: Synthesis of 4-(5-bromo-4-(isopropylamino)-3-nitropyridin-2-yl)-3-fluorobenzonitrile (A-3)

5-Bromo-2-chloro-N-isopropyl-3-nitropyridine-4-amine (A-2, 3.44 g, 11.68 mmol) and 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (2.89 g, 11.68 mmol) were dissolved in a mixed solvent of 1,4-dioxane (30 mL) and water (3 mL), tetrakis(triphenylphosphine)palladium (1.35 g, 1.17 mmol) and sodium bicarbonate (1.96 g, 23.36 mmol) were added, and the mixture was heated to 100°C under argon protection and reacted overnight. LC-MS detection showed that the reaction was complete. The reaction solution was cooled to room temperature, diluted with ethyl acetate (30 mL) and filtered. The filtrate was washed with saturated brine (30 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 0-20% gradient elution) to give the title compound (1.66 g).

MS m/z(ESI):379.0,381.1[M+H] ⁺ .

Step 3: Synthesis of 4-(3-amino-5-bromo-4-(isopropylamino)pyridin-2-yl)-3-fluorobenzonitrile (A-4)

4-(5-bromo-4-(isopropylamino)-3-nitropyridin-2-yl)-3-fluorobenzonitrile (A-3, 1.66 g, 4.38 mmol) was dissolved in a mixed solvent of ethanol (50 mL) and water (5 mL), and ammonium chloride (2.34 g, 43.78 mmol) and reduced iron powder (1.22 g, 21.89 mmol) were added, and the temperature was raised to 60°C for overnight reaction. LC-MS detection showed that the reaction was complete. The reaction solution was cooled to room temperature, diluted with ethyl acetate (50 mL) and filtered, and the filtrate was washed with saturated brine (20 mL × 3), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 0-30% gradient elution) to obtain the title compound (1.06 g).

MS m/z(ESI):349.0,351.0[M+H] ⁺ .

Step 4: Synthesis of 3-bromo-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (A-5)

4-(3-amino-5-bromo-4-(isopropylamino)pyridin-2-yl)-3-fluorobenzonitrile (A-4, 1.06 g, 3.04 mmol) was dissolved in N,N-dimethylformamide (20 mL), cesium carbonate (2.97 g, 9.11 mmol) was added, and then the temperature was raised to 100°C for 6 h. LC-MS detection showed that the reaction was complete. The reaction solution was cooled to room temperature, diluted with ethyl acetate (30 mL) and filtered. The filtrate was washed with saturated brine (20 mL*3), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 0-30% gradient elution) to obtain the title compound (361 mg).

MS m/z(ESI):329.0,331.0[M+H] ⁺ .

Step 5: Synthesis of tert-butyl 4-(5-(7-cyano-4-(isopropylamino)-5H-pyrido[3,2-b]indol-3-yl)-1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (A-6)

3-Bromo-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (A-5, 100 mg, 304 μmol) and tert-butyl 4-(1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (164 mg, 608 μmol) were dissolved in 1,4-dioxane (3 mL), palladium acetate (7 mg, 30 μmol), cuprous iodide (12 mg, 61 μmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (35 mg, 61 μmol) and cesium carbonate (297 mg, 911 μmol) were added and the temperature was raised to 100 °C under argon protection to react overnight. LC-MS detection showed that the reaction was complete. The reaction solution was cooled to room temperature, diluted with dichloromethane (10 mL), washed with saturated brine (10 mL*3), and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (methanol/dichloromethane=0-10% gradient elution) to obtain the title compound (31 mg).

MS m/z(ESI):519.2[M+H] ⁺ .

Step 6: Synthesis of 4-(isopropylamino)-3-(5-(piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-5H-pyrido[3,2-b]indole-7-carbonitrile (Intermediate A)

Tert-butyl 4-(5-(7-cyano-4-(isopropylamino)-5H-pyrido[3,2-b]indol-3-yl)-1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (A-6, 26 mg, 50 μmol) was dissolved in 1,4-dioxane (1 mL), and a hydrogen chloride-1,4-dioxane solution (4 M, 0.5 mL) was added and the reaction was continued at room temperature for 1 h. LC-MS detection showed that the reaction was complete, and the reaction solution was concentrated to dryness under reduced pressure to obtain a crude product of the title compound (30 mg).

MS m/z(ESI):419.2[M+H] ⁺ .

Intermediate B: Synthesis of 4-(isopropylamino)-3-[1-(piperidin-4-yl)-1H-1,2,3-triazol-4-yl]-5H-pyrido[3,2-b]indole-7-carbonitrile

Step 1: Synthesis of 4-[5-[2-[tert-butyl(dimethyl)silyl]ethynyl]-4-(isopropylamino)-3-nitro-pyridin-2-yl]-3-fluoro-benzonitrile (B-1)

4-(5-bromo-4-(isopropylamino)-3-nitropyridin-2-yl)-3-fluorobenzonitrile (A-3, 750 mg, 1.98 mmol) was dissolved in 1,4-dioxane (10 mL), tert-butylethynyldimethylsilane (555 mg, 3.96 mmol), cuprous iodide (75.3 mg, 0.39 mmol) and cesium carbonate (1.29 g, 3.96 mmol) were added, and the mixture was heated to 100°C under argon protection and reacted overnight. LC-MS detection showed that the reaction was complete. The reaction solution was diluted with ethyl acetate (20 mL), and then washed with saturated brine (20 mL × 3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 0-10% gradient elution) to obtain the title compound (670 mg).

MS m/z(ESI):439.2[M+H] ⁺ .

Step 2: Synthesis of 4-[3-amino-5-[2-[tert-butyl(dimethyl)silyl]ethynyl]-4-(isopropylamino)-pyridin-2-yl]-3-fluoro-benzonitrile (B-2)

Dissolve 4-[5-[2-[tert-butyl(dimethyl)silyl]ethynyl]-4-(isopropylamino)-3-nitro-pyridin-2-yl]-3-fluoro-benzonitrile (B-1, 500 mg, 1.14 mmol) in a mixed solvent of ethanol (10 mL) and water (4 mL), add ammonium chloride (610 mg, 11.4 mmol) and reduced iron powder (318 mg, 5.7 mmol), and heat to 60 ° C to react overnight. LC-MS detection shows that the reaction is complete. The reaction solution is cooled to room temperature, filtered, and the filtrate is removed by rotation and extracted with ethyl acetate (20 mL) and water (20 mL). The organic phase is washed with saturated brine (10 mL×2), dried over anhydrous sodium sulfate, and filtered. The filtrate is concentrated to dryness under reduced pressure to obtain the title compound (460 mg).

MS m/z(ESI):409.2[M+H] ⁺ .

Step 3: Synthesis of 3-ethynyl-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (B-3)

4-[3-amino-5-[2-[tert-butyl(dimethyl)silyl]ethynyl]-4-(isopropylamino)-pyridin-2-yl]-3-fluoro-benzonitrile (B-2, 160 mg, 0.39 mmol) was dissolved in N,N-dimethylformamide (6 mL), cesium carbonate (383 mg, 1.17 mmol) was added, and then the temperature was raised to 100°C for 12 h. LC-MS detection showed that the reaction was complete. The reaction solution was cooled to room temperature, diluted with ethyl acetate (10 mL) and filtered. The filtrate was washed with saturated brine (10 mL×3), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate/petroleum ether = 0-50% gradient elution) to obtain the title compound (14.0 mg).

MS m/z(ESI):275.1[M+H] ⁺ .

Step 4: Synthesis of tert-butyl 4-[4-[7-cyano-4-(isopropylamino)-5H-pyrido[3,2-b]indol-3-yl]-1H-1,2,3-triazol-1-yl]piperidine-1-carboxylate (B-4)

3-Ethynyl-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (B-3, 8.0 mg, 29 μmol) and tert-butyl 4-azidopiperidine-1-carboxylate (6.6 mg, 29 μmol) were dissolved in tetrahydrofuran (0.5 mL), and cuprous thiophene-2-carboxylate (0.55 mg, 2.9 μmol) was added and reacted at room temperature overnight under argon protection. LC-MS detection showed that the reaction was complete, and the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (methanol/dichloromethane = 0-10% gradient elution) to obtain the title compound (4.0 mg).

MS m/z(ESI):501.3[M+H] ⁺ .

Step 5: Synthesis of 4-(isopropylamino)-3-[1-(piperidin-4-yl)-1H-1,2,3-triazol-4-yl]-5H-pyrido[3,2-b]indole-7-carbonitrile (Intermediate B)

4-[4-[7-cyano-4-(isopropylamino)-5H-pyrido[3,2-b]indol-3-yl]-1H-1,2,3-triazole-triazol-1-yl]piperidine-1-carboxylic acid tert-butyl ester (B-4, 4.0 mg, 7.9 μmol) was dissolved in ethyl acetate (0.5 mL), and hydrogen chloride-1,4-dioxane solution (4M, 0.02 mL) was added and the reaction was continued at room temperature for 2 h. LC-MS detection showed that the reaction was complete, and the reaction solution was concentrated to dryness under reduced pressure to obtain the title compound (3.5 mg).

MS m/z (ESI): 401.2 [M+H] ⁺ ;

¹ H NMR (400MHz, D ₂ O)δ8.51(s,1H),8.05(s,1H),7.82-7.80(m,1H),7.70(s,1H),7.20-7.18(m,1H),5.01-4.99(m,1H),4.21-4 .18(m,1H),3.67-3.63(m,2H),3.33-3.27(m,2H),2.58-2.55(m,2H),2.43-2.37(m,2H),1.38-1.36(m,6H).

Intermediate C: Synthesis of 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxaldehyde

Step 1: Synthesis of 2,6-bis(benzyloxy)-6'-fluoro-3,3'-bipyridine (C-2)

2-Fluoro-5-iodopyridine (C-1, 3 g, 13.5 mmol) and 2,6-dibenzyloxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (8.46 g, 20.3 mmol) were dissolved in 1,4-dioxane (80 mL), followed by the addition of water (20 mL), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (495 mg, 0.68 mmol) and cesium carbonate (6.60 g, 20.27 mmol), and stirred at 90° C. After LC-MS showed that the reaction was complete, water (200 mL) was added, and the mixture was extracted with ethyl acetate (40 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=5/1) to obtain the title compound (3.5 g).

MS m/z[ESI]:387.10[M+H] ⁺ .

Step 2: Synthesis of 3-(6-fluoropyridin-3-yl)piperidine-2,6-dione (C-3)

Dissolve 2,6-bis(benzyloxy)-6'-fluoro-3,3'-bipyridine (C-2, 3.5 g, 9.06 mmol) in ethyl acetate (100 mL), add palladium carbon (2.0 g), and stir under a hydrogen atmosphere. Stir the reaction at 50 ° C for 12 h. After LC-MS showed that the reaction was complete, filter and concentrate the filtrate under reduced pressure to obtain the crude title compound (1.8 g), which was directly used for the next step without purification.

MS m/z[ESI]:209.1[M+H] ⁺ .

Step 3: Synthesis of 3-(6-(4-(dimethoxymethyl)piperidin-1-yl)pyridin-3-yl)piperidine-2,6-dione (C-4)

3-(6-fluoropyridin-3-yl)piperidine-2,6-dione (C-3, 450 mg, 2.16 mmol) and 4-(dimethoxymethyl)piperidine (1.03 mg, 6.48 mmol) were dissolved in dimethyl sulfoxide (5 mL), N,N-diisopropylethylamine (558.70 mg, 4.32 mmol) was added, and the temperature was raised to 120°C for microwave reaction for 2.5 h. After LC-MS showed that the reaction was complete, 100 mL of ethyl acetate was added for dilution, and the organic phase was washed with water (30 mL) and saturated brine (30 mL), and dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/ethyl acetate = 1/1) to obtain the title compound (340 mg).

MS m/z(ESI):348.2[M+H] ⁺ .

Step 4: Synthesis of 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carbaldehyde (Intermediate C)

3-(6-(4-(dimethoxymethyl)piperidin-1-yl)pyridin-3-yl)piperidine-2,6-dione (C-4, 40 mg, 115.14 μmol) was dissolved in tetrahydrofuran (2 mL), diluted hydrochloric acid (2N, 575 μL) was added, and the temperature was raised to 60°C for 2 h. After the reaction was completed, the reaction solution was directly lyophilized to obtain the title compound (40.0 mg).

MS m/z(ESI):302.1[M+H] ⁺ .

Intermediate D: Synthesis of 1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidine-4-carboxaldehyde

At room temperature, 5-(4-(dimethoxymethyl)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (C-1, 30 mg, 0.072 mmol) was dissolved in acetone (1 mL). Aqueous hydrochloric acid solution (2 mL, 1 N) was added dropwise at room temperature, and the mixture was heated to 80°C and stirred for 1 hour. The reaction was complete after TLC monitoring. Saturated aqueous sodium bicarbonate solution (20 mL) was added for dilution, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined and concentrated in vacuo to obtain the crude title compound, which was directly used in the next step.

MS m/z(ESI):370.1[M+H] ⁺ .

Intermediate E: Synthesis of 1-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)piperidine-4-carboxaldehyde

Step 1: Synthesis of 3-(6-(4-(dimethoxymethyl)piperidin-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (E-2)

At room temperature, 3-(6-bromo-1-oxo-isoindolin-2-yl)piperidine-2,6-dione (E-1, 800 mg, 2.48 mmol) and 4-(dimethoxymethyl)piperidine (394.19 mg, 2.48 mmol) were dissolved in dioxane (10 mL), (SP-4-1)-[1,3-bis[2,6-bis(1-ethylpropyl)phenyl]-4,5-dichloro-1,3-dihydro-2H-imidazol-2-ylidene]dichloro(2-methylpyridine)palladium (145.57 mg, 173.30 μmol) and cesium carbonate (2.42 g, 7.43 mmol) were added, N ₂ was replaced, the temperature was raised to 100° C., and the reaction mixture was stirred at 100° C. for 16 hours. After the reaction is completed, dilute with dichloromethane, add 5% acetic acid solution to adjust the pH to 6-7, extract with dichloromethane, dry the organic phase with saturated brine and anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain a crude product, which is separated by column chromatography (dichloromethane: methanol = 15: 1) to obtain the title compound (468.4 mg).

MS m/z(ESI):402.1[M+H] ⁺

Step 2: Synthesis of 1-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)piperidine-4-carbaldehyde (Intermediate E)

At room temperature, 3-(6-(4-(dimethoxymethyl)piperidin-1-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (E-2, 37 mg, 0.092 mmol) was dissolved in acetone (1 mL). Aqueous hydrochloric acid solution (2 mL, 1 N) was added dropwise at room temperature, and the mixture was heated to 80°C and stirred for 1 hour. The reaction was monitored by a plate. Saturated aqueous sodium bicarbonate solution (20 mL) was added for dilution, and the mixture was extracted with dichloromethane (15 mL*3). The organic phases were combined and concentrated in vacuo to obtain the crude title compound (33 mg), which was used directly in the next step.

Intermediate F: Synthesis of 1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)piperidine-4-carboxaldehyde

Referring to the synthesis method of intermediate E, intermediate E-1 was replaced with 3-(5-bromo-1-oxo-isoindolin-2-yl)piperidine-2,6-dione, and intermediate F was prepared in the same manner.

Intermediate G: Synthesis of 1-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)piperidine-4-carboxaldehyde

Step 1: Synthesis of 3-(5-(4-(dimethoxymethyl)piperidin-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (G-2)

Under nitrogen protection, 3-(5-bromo-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (G-1, 500 mg, 1.48 mmol) and 4-(dimethoxymethyl)piperidine (470.86 mg, 2.96 mmol) were dissolved in dioxane (6 mL), dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (144 mg, 147.86 μmol) and cesium carbonate (2.41 g, 7.39 mmol) were added thereto, and the reaction was stirred at 100°C for 6 hours. LCMS showed that the reaction was complete. Ethyl acetate (10 mL) and water (10 mL×2) were added for extraction, the organic phases were combined, the organic phases were dried, filtered, and the filtrate was concentrated to obtain the title compound (20 mg).

Step 2: Synthesis of 1-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)piperidine-4-carbaldehyde (Intermediate G)

3-(5-(4-(dimethoxymethyl)piperidin-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione (G-2, 18 mg, 43.22 μmol) was dissolved in a mixed solution of tetrahydrofuran (2 mL) and water (0.4 mL), and hydrochloric acid (2M, 216.10 μL) was added, and the mixture was reacted at 60°C for 3 hours. The solvent was removed by concentration under reduced pressure, and appropriate amounts of acetonitrile and water were added and freeze-dried to obtain the title compound (15 mg).

MS m/z(ESI):371.00[M+H] ⁺

Intermediate H: Synthesis of 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2-azaspiro[3.5]nonane-7-carbaldehyde

Step 1: Synthesis of (2-azaspiro[3.5]nonan-7-yl)methanol (H-2)

At room temperature, tert-butyl 7-(hydroxymethyl)-2-azaspiro[3.5]nonane-2-carboxylate (H-1, 800 mg, 3.13 mmol) was dissolved in dichloromethane (4 mL). Hydrochloric acid dioxane solution (2 mL, 4 N) was added dropwise at room temperature and stirred at room temperature for 2 hours. The reaction solution was concentrated in vacuo to remove the solvent to obtain the crude title compound, which was used directly in the next step.

Step 2: Synthesis of 2-(2,6-dioxopiperidin-3-yl)-5-(7-(hydroxymethyl)-2-azaspiro[3.5]nonan-2-yl)isoindoline-1,3-dione (H-3)

2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (864.6 mg, 3.13 mmol), (2-azaspiro[3.5]nonane-7-yl)methanol (H-2, crude product, 3.13 mmol) and diisopropylethylamine (2.02 g, 15.65 mmol) were dissolved in dimethyl sulfoxide (5 mL), the temperature was raised to 100°C, and the reaction mixture was stirred at 100°C for 4 hours. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with water (100 mL), filtered, the filter cake was collected, and separated by silica gel column chromatography (dichloromethane/methanol = 100/1 to 100/2) to obtain the title compound (1.20 g).

MS m/z(ESI):412.2[M+H] ⁺

Step 3: Synthesis of 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2-azaspiro[3.5]nonane-7-carbaldehyde (Intermediate H)

Under ice bath, 2-(2,6-dioxopiperidin-3-yl)-5-(7-(hydroxymethyl)-2-azaspiro[3.5]nonane-2-yl)isoindoline-1,3-dione (H-3, 500.0 mg, 1.22 mmol) was dissolved in dichloromethane (10 mL), and Dess-Martin periodinane (776.2 mg, 1.83 mmol) was slowly added, and the reaction mixture was stirred at 0°C for 1 hour, warmed to room temperature, and stirred for 2 hours. After the reaction was completed, it was concentrated and separated and purified by silica gel column chromatography (dichloromethane/methanol=100/1-100/2) to obtain the title compound (450.0 mg).

MS m/z(ESI):410.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ ) δ 11.06 (s, 1H), 9.59 (s, 1H), 7.63 (d, J = 8.4Hz, 1H), 6.76 (d, J = 2.0Hz, 1H), 6.63 (dd, J ₁ = 2.0Hz, J ₂ = 8.0Hz, 1H), 5.05 (dd, J ₁ =5.6Hz,J ₂ ＝12.8Hz,1H),3.72(s,4H),2.93-2.83(m,1H),2.60-2.47(m,2H),2.35-2.29(m, 1H),2.03-1.97(m,1H),1.91-1.80(m,4H),1.63-1.47(m,2H),1.43-1.34(m,2H).

Intermediate I: Synthesis of 1-(4-((2,6-dioxopiperidin-3-yl)oxy)phenyl)piperidine-4-carboxaldehyde

Step 1: Synthesis of 1-(4-benzyloxyphenyl)-4-(dimethoxymethyl)piperidine (I-2)

1-(Benzyloxy)-4-iodobenzene (I-1, 3 g, 9.67 mmol), 4-(dimethoxymethyl)piperidine (4.62 g, 29.02 mmol), 2-(di-tert-butylphosphine)biphenyl (577.31 mg, 1.93 mmol), potassium tert-butoxide (3.26 g, 29.02 mmol) and tris(dibenzylideneacetone)dipalladium (885.81 mg, 967.34 μmol) were dissolved in tert-amyl alcohol (40 mL) and stirred at 100 °C overnight. After LC-MS showed that the reaction was complete, saturated chlorine was added. Add ammonium hydroxide aqueous solution (50 mL), extract with ethyl acetate (50 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter and concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether/ethyl acetate=5/1) to give the title compound (2.14 g).

MS m/z(ESI):342.2[M+H] ⁺

Step 2: Synthesis of 4-(4-(dimethoxymethyl)piperidin-1-yl)phenol (I-3)

1-(4-Benzyloxyphenyl)-4-(dimethoxymethyl)piperidine (I-2, 1.99 g, 5.83 mmol) was dissolved in ethyl acetate (40 mL), 10% palladium on carbon (55% wt H ₂ O, 400 mg) was added, and the mixture was reacted at 60° C. overnight under hydrogen protection. After LC-MS showed that the reaction was complete, the reaction solution was diluted with ethyl acetate (60 mL) and filtered through diatomaceous earth, the filter cake was rinsed with ethyl acetate (20 mL), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=4/1) to obtain the title compound (1.43 g).

MS m/z(ESI):252.2[M+H] ⁺

Step 3: Synthesis of 3-(4-(4-(dimethoxymethyl)piperidin-1-yl)phenoxy)piperidine-2,6-dione (I-4)

4-(4-(dimethoxymethyl)piperidin-1-yl)phenol (I-3, 1.38 g, 5.49 mmol) was dissolved in tetrahydrofuran (20 mL), sodium hydride (329.46 mg, 8.24 mmol, purity 60%) was added under ice bath, 3-bromopiperidine-2,6-dione (1.37 g, 7.14 mmol) was added after stirring for 15 minutes, and the reaction was stirred at room temperature overnight. After LC-MS showed that the reaction was complete, the reaction solution was added dropwise to saturated aqueous ammonium chloride solution (40 mL) to quench the residual sodium hydrogen, and then extracted with ethyl acetate (30 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=2/3) to obtain the title compound (1.62 g).

MS m/z(ESI):363.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.88(s,1H),6.94-6.83(m,4H),5.02-4.97(m,1H),4.09(d,J＝6.7Hz,1H),3.58-3.49(m,2H),3.28( s,6H),2.73-2.64(m,1H),2.56-2.51(m,4H),2.23-2.03(m,2H),1.75-1.67(m,2H),1.41-1.23(m,2H).

Step 4: Synthesis of 1-(4-((2,6-dioxopiperidin-3-yl)oxy)phenyl)piperidine-4-carbaldehyde (Intermediate I)

At room temperature, 3-(4-(4-(dimethoxymethyl)piperidin-1-yl)phenoxy)piperidine-2,6-dione (50 mg, 0.138 mmol) was dissolved in acetone (1 mL). Aqueous hydrochloric acid solution (2 mL, 1 N) was added dropwise at room temperature, and the mixture was heated to 80°C and stirred for 1 hour. The reaction was complete after TLC monitoring. Saturated aqueous sodium bicarbonate solution (20 mL) was added for dilution, and the mixture was extracted with dichloromethane (15 mL × 3). The organic phases were combined and concentrated in vacuo to obtain the crude title compound, which was directly used in the next step.

Intermediate L: Synthesis of 4-(isopropylamino)-3-(5-(piperazin-1-yl)-1,3,4-thiadiazol-2-yl)benzofurano[3,2-b]pyridine-7-carbonitrile

Step 1: Synthesis of 4-bromo-3-((2-chloro-4-nitropyridin-3-yl)oxy)benzonitrile (L-2)

4-Bromo-3-hydroxybenzonitrile (L-1, 10 g, 50.50 mmol) was dissolved in tetrahydrofuran (100 mL), sodium hydrogen (4.04 g, 101.00 mmol, purity 60%) was added under ice bath, stirred for 30 minutes, then 2-chloro-3-fluoro-4-nitro-pyridine (8.92 g, 50.50 mmol) was added and stirred at room temperature for 2 hours, and the reaction was monitored by LCMS. After the reaction was completed, the reaction solution was diluted with ethyl acetate (10 mL × 3), washed with saturated brine (30 mL) under water, the organic phase was dried, concentrated to dryness, and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain the title compound (15.38 g).

MS m/z(ESI):355.9[M+H] ⁺

Step 2: Synthesis of 3-((4-amino-2-chloropyridin-3-yl)oxy)-4-bromobenzonitrile (L-3)

4-Bromo-3-((2-chloro-4-nitropyridin-3-yl)oxy)benzonitrile (L-2, 15.59 g, 43.97 mmol) was dissolved in ethanol (15 mL) and water (6 mL), and iron powder (12.28 g, 219.86 mmol) and ammonium chloride (23.52 g, 439.72 mmol) were added, and stirred at 60°C overnight. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, extracted with ethyl acetate (30 mL×3) and water (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (dichloromethane/methanol=20/1) to obtain the title compound (5.61 g).

MS m/z(ESI):325.9[M+H] ⁺

Step 3: Synthesis of 3-((4-amino-2-chloropyridin-3-yl)oxy)-4-(trimethylstannyl)benzonitrile (L-4)

3-((4-amino-2-chloropyridin-3-yl)oxy)-4-bromobenzonitrile (L-3, 1.36 g, 4.19 mmol) was dissolved in 1,4-dioxane (30 mL), tetrakistriphenylphosphine palladium (484.21 mg, 419.03 μmol) and hexamethyltin (1.51 g, 4.61 mmol) were added, and the mixture was reacted at 100°C overnight under argon protection. LC-MS detection showed that the reaction was complete, and the reaction solution was added with saturated sodium chloride aqueous solution (30 mL), extracted with ethyl acetate (30 mL×3), the organic phases were combined, dried with anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=2/1) to obtain the title compound (1.5 g).

MS m/z(ESI):410.0[M+H] ⁺

Step 4: Synthesis of 4-aminobenzofurano[3,2-b]pyridine-7-carbonitrile (L-5)

3-((4-amino-2-chloropyridin-3-yl)oxy)-4-(trimethyltinyl)benzonitrile (L-4, 1.4 g, 3.43 mmol) was dissolved in 1,4-dioxane (50 mL), and methanesulfonic acid (2-dicyclohexylphosphine-2',4',6'-triisopropyl-1,1'-biphenyl) (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (580.23 mg, 685.49 μmol) was added, and the mixture was reacted at 100°C overnight under argon protection. LC-MS detection showed that the reaction was complete, and the reaction solution was added with saturated sodium chloride aqueous solution (50 mL), extracted with ethyl acetate (50 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (dichloromethane/methanol=10/1) to obtain the title compound (423 mg).

MS m/z(ESI):210.0[M+H] ⁺

Step 5: Synthesis of 4-aminobenzofurano[3,2-b]pyridine-7-carbonitrile (L-6)

4-Aminobenzofurano[3,2-b]pyridine-7-carbonitrile (L-5, 370 mg, 1.77 mmol) was dissolved in acetonitrile (10 mL), acetic acid (106 mg, 1.77 mmol) and N-iodosuccinimide (796 mg, 3.54 mmol) were added, and stirred at 70°C for 12 hours. The solvent was removed by concentration under reduced pressure, ethyl acetate (5 mL×3) and water (10 mL) were added for extraction, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/methanol=20/1) to obtain the title compound (439 mg).

MS m/z(ESI):336.1[M+H] ⁺

Step 6: Synthesis of 3-iodo-4-(isopropylamino)benzofurano[3,2-b]pyridine-7-carbonitrile (L-7)

4-Aminobenzofurano[3,2-b]pyridine-7-carbonitrile (L-6, 503 mg, 1.50 mmol) was dissolved in N,N-dimethylformamide (6 mL), sodium hydrogen (180 mg, 4.50 mmol, purity 60%) was added to the solution at 0 ° C, and after the reaction solution was warmed to room temperature, N,N-dimethylformamide solution containing 2-iodopropane (766 mg, 4.51 mmol) was added dropwise at 80 ° C, and the reaction was carried out for 6 hours. LCMS showed that the main product was the product. The reaction solution was cooled to room temperature and quenched with crushed ice. The product was extracted with dichloromethane (2×20 mL) and water (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 10/1) to obtain the title compound (71 mg).

MS m/z(ESI):378.18[M+H] ⁺

Step 7: Synthesis of tert-butyl 4-(5-(7-cyano-4-(isopropylamino)benzofurano[3,2-b]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (L-8)

Under nitrogen atmosphere, 3-iodo-4-(isopropylamino)benzofurano[3,2-b]pyridine-7-carbonitrile (L-7, 60 mg, 159.08 μmol) and tert-butyl 4-(1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (43 mg, 159.08 μmol) were dissolved in dioxane (0.5 mL), cesium carbonate (207 mg, 636.30 μmol), cuprous iodide (6 mg, 31.82 μmol), palladium acetate (7 mg, 31.18 μmol) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (37 mg, 63.95 μmol) and trimethylacetic acid (5 mg, 48.96 μmol) were added, and the mixture was heated to 100° C. and stirred for 12 hours. LCMS showed that the mixture was mainly the product. The solvent was removed by concentration under reduced pressure, and the residue was extracted with ethyl acetate (5 mL×3) and water (20 mL). The residue was purified by thin layer chromatography (dichloromethane/methanol=10/1) twice to give the title compound (26 mg).

MS m/z(ESI):520.6[M+H] ⁺

Step 8: Synthesis of 4-(isopropylamino)-3-(5-(piperazin-1-yl)-1,3,4-thiadiazol-2-yl)benzofurano[3,2-b]pyridine-7-carbonitrile (Intermediate L)

4-[5-[7-cyano-4-(isopropylamino)benzofurano[3,2-b]pyridin-3-yl]-1,3,4-thiadiazol-2-yl]piperazine-1-carboxylic acid tert-butyl ester (L-8, 20 mg, 38.49 μmol) was dissolved in a mixed solution of dichloromethane (3 mL) and trifluoroacetic acid (0.6 mL) and stirred at room temperature for 2 hours. LCMS showed that it was mainly the product. The solvent was removed by concentration to obtain the title compound (16 mg).

MS m/z(ESI):420.10[M+H] ⁺

Intermediate M: Synthesis of 1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-6-yl)piperidine-4-carboxaldehyde

Step 1: Synthesis of 6-bromo-3-iodo-1-methyl-1H-indazole (M-2)

The compound 6-bromo-3-iodo-1H-indazole (M-1, 5.0 g, 15.5 mmol) and potassium hydroxide (2.17 g, 38.71 mmol) were dissolved in acetone (30 mL), the temperature was lowered to 0°C, methyl iodide (3.30 g, 23.22 mmol) was added to the reaction solution, and the reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction solution was filtered and the filter cake was concentrated under reduced pressure. Water (30 mL) was added for dilution and ethyl acetate (50 mL × 3) was used for extraction. The organic phase was washed with saturated brine (30 mL × 2), separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain the title compound (3.15 g).

MS m/z(ESI):336.9[M+H] ⁺

Step 2: Synthesis of 3-(2,6-di(benzyloxy)pyridin-3-yl)-6-bromo-1-methyl-1H-indazole (M-3)

At room temperature, compound 6-bromo-3-iodo-1-methyl-1H-indazole (M-2, 3.15 g, 9.35 mmol), 2,6-di(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (3.48 g, 8.35 mmol), cesium carbonate (5.44 g, 16.69 mmol) and 1,1'-di(diphenylphosphino)ferrocenepalladium dichloride (0.18 g, 0.25 mmol) were dissolved in tetrahydrofuran/water (90 mL/15 mL), nitrogen was protected, the temperature was raised to 80°C, and the reaction mixture was stirred at 80°C for 14 hours. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with water (30 mL), and extracted with ethyl acetate (30 mL×3). The organic phase was washed with saturated brine (20 mL x 2), separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain the title compound (2.9 g).

Step 3: Synthesis of 3-(2,6-di(benzyloxy)pyridin-3-yl)-6-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazole (M-4)

At room temperature, compound 3-(2,6-di(benzyloxy)pyridin-3-yl)-6-bromo-1-methyl-1H-indazole (M-3, 2.9 g, 5.81 mmol), 4-(dimethoxymethyl)piperidine (1.87 g, 11.77 mmol), cesium carbonate (5.75 g, 17.64 mmol), 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (0.27 g, 0.59 mmol) and methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium (II) (0.49 g, 0.59 mmol) were dissolved in toluene (40 mL). Under nitrogen protection, the temperature was raised to 100°C and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated under reduced pressure, diluted with water (30 mL), and extracted with ethyl acetate (3 0 mL × 3). The organic phase was washed with saturated brine (20 mL × 2), separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain the title compound (2.07 g).

MS m/z(ESI):579.4[M+H] ⁺

Step 4: Synthesis of 3-(6-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazol-3-yl)piperidine-2,6-dione (M-5)

At room temperature, 3-(2,6-di(benzyloxy)pyridin-3-yl)-6-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazole (M-4, 717 mg, 1.24 mmol) was dissolved in tetrahydrofuran/anhydrous ethanol (7 mL/7 mL), followed by the addition of palladium carbon (70 mg) and palladium hydroxide (70 mg). The reaction mixture was stirred at room temperature for 12 hours under a hydrogen atmosphere (0.1 MPa). The reaction solution was filtered through a pad of celite, and the filtrate was concentrated under reduced pressure to obtain the title compound (471 mg).

MS m/z(ESI):401.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.85(s,1H),7.47(d,J＝8.8Hz,1H),6.90(dd,J＝1.2Hz,9.2Hz,1H),6.82(s,1H),4.26-4.24(m,1H),4.10(d,J＝6.4Hz,1H),3. 88(s,3H),3.80(d,J＝12.4Hz,2H),3.28(s,6H),2.71-2.59(m,4H),2.30-2.14(m,2H),1.73(d,J＝10.4Hz,3H),1.38-1.36(m,2H).

Step 5: Synthesis of 1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-6-yl)piperidine-4-carbaldehyde (Intermediate M)

At room temperature, 3-(6-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazol-3-yl)piperidine-2,6-dione (M-5, 40 mg, 0.100 mmol) was dissolved in acetone (1 mL). Aqueous hydrochloric acid solution (2 mL, 1 N) was added dropwise at room temperature, and the mixture was heated to 80°C and stirred for 1 hour. The reaction was monitored by TLC to be complete. Saturated aqueous sodium bicarbonate solution (20 mL) was added for dilution, and the mixture was extracted with dichloromethane (15 mL×3). The organic phases were combined and concentrated in vacuo to obtain the crude title compound, which was directly used in the next step.

Intermediate N: Synthesis of 1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)piperidine-4-carboxaldehyde

Step 1: Synthesis of 7-bromo-3-iodo-1H-indazole (N-2)

At room temperature, 7-bromo-1H-indazole (N-1,5 g, 25.38 mmol) was dissolved in N,N-dimethylformamide (60 mL), potassium hydroxide (2.15 g, 38.27 mmol) and iodine (9.71 g, 38.27 mmol) were added at 0 ° C. After the reaction solution was stirred at room temperature for 16 h, it was quenched with water (100 mL), and then quenched with saturated sodium sulfite aqueous solution (100 mL) and stirred for 15 min. The reaction mixture was filtered and the filter was washed with water to obtain the title compound (8 g).

MS m/z(ESI):322.9[M+H] ⁺

Step 2: Synthesis of 7-bromo-3-iodo-1-methyl-1H-indazole (N-3)

At room temperature, 7-bromo-3-iodo-1H-indazole (N-2, 8g, 24.8mmol) was dissolved in THF (150mL), and a 1M THF solution of potassium tert-butoxide (49.70mL, 49.70mmol) was added dropwise at 0°C for 1 hour. At 0°C, a solution of iodomethane (6.35g, 44.72mmol) in tetrahydrofuran (50mL) was added dropwise. After addition, the reaction mixture was stirred at 15°C for 14 hours. The reaction solution was filtered and concentrated, and the crude product was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1) to obtain the title compound (5.4g).

MS m/z(ESI):336.9[M+H] ⁺

Step 3: Synthesis of 3-(2,6-di(benzyloxy)pyridin-3-yl)-7-bromo-1-methyl-1H-indazole (N-4)

At room temperature, 7-bromo-3-iodo-1-methyl-1H-indazole (N-3,3g, 8.9mmol), 2,6-di(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (4.09g, 9.8mmol), potassium carbonate (2.46g, 17.8mmol) and 1,1'-di(diphenylphosphino)ferrocenepalladium dichloride (653.68mg, 0.893mmol) were dissolved in 1,4-dioxane/water (30mL/3mL), nitrogen was replaced, the temperature was raised to 90℃, and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with water (50mL), and extracted with ethyl acetate (50mL×3). The organic phase was washed with saturated brine (20mL×2), separated, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate=5/1) to give the title compound (2.58 g).

MS m/z(ESI):500.1[M+H] ⁺

Step 4: Synthesis of 3-(2,6-di(benzyloxy)pyridin-3-yl)-7-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazole (N-5)

At room temperature, 3-(2,6-di(benzyloxy)pyridin-3-yl)-7-bromo-1-methyl-1H-indazole (N-4, 2.58 g, 5.16 mmol), 4-(dimethoxymethyl)piperidine (1.64 g, 10.32 mmol), cesium carbonate (5.05 g, 15.48 mmol), 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (241 mg, 0.516 mmol) and methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium (II) (452 mg, 0.540 mmol) were dissolved in 1,4-dioxane (40 mL), nitrogen was replaced, the temperature was raised to 100°C, and the reaction mixture was stirred at 100°C for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated under reduced pressure, diluted with water (30 mL), and extracted with ethyl acetate (30 mL × 3). The organic phase was washed with saturated brine (20 mL × 2), separated, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate = 3/1) to obtain the title compound (1.16 g).

MS m/z(ESI):579.4[M+H] ⁺

Step 5: Synthesis of 3-(7-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazol-3-yl)piperidine-2,6-dione (N-6)

At room temperature, 3-(7-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazol-3-yl)piperidine-2,6-dione (N-5, 980 mg, 1.69 mmol) was dissolved in tetrahydrofuran/anhydrous ethanol (7 mL/7 mL), followed by addition of palladium carbon (90 mg) and palladium hydroxide (90 mg), hydrogen replacement, and the reaction mixture was stirred at 25°C for 16 hours. The reaction solution was filtered through a pad of celite, and the filtrate was concentrated under reduced pressure to obtain the title compound (590 mg).

MS m/z(ESI):401.1[M+H] ⁺

Step 6: Synthesis of 1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)piperidine-4-carbaldehyde (Intermediate N)

At room temperature, 3-(7-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazol-3-yl)piperidine-2,6-dione (N-6, 37 mg, 0.092 mmol) was dissolved in acetone (1 mL). Aqueous hydrochloric acid solution (2 mL, 1 N) was added dropwise at room temperature, and the mixture was heated to 80°C and stirred for 1 hour. The reaction was monitored by TLC to be complete. Saturated aqueous sodium bicarbonate solution (20 mL) was added for dilution, and the mixture was extracted with dichloromethane (15 mL*3). The organic phases were combined and concentrated in vacuo to obtain the crude title compound, which was used directly in the next step.

Synthesis of Intermediate O: N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(4-formylpiperidin-1-yl)benzamide

Step 1: Synthesis of methyl 4-(4-(dimethoxymethyl)piperidin-1-yl)-2-fluorobenzoate (O-2)

At room temperature, methyl 4-bromo-2-fluorobenzoate (O-1, 2.0 g, 8.58 mmol) and 4-(dimethoxymethyl)piperidine (2.05 g, 12.87 mmol) were dissolved in 1,4-dioxane (30 mL), cesium carbonate (5.58 g, 17.16 mmol), 2-dicyclohexylphosphino-2',6'-diisopropoxybiphenyl (401 mg, 0.86 mmol) and methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium (II) (719 mg, 0.86 mmol) were added, the reaction system was replaced with nitrogen three times, and the reaction mixture was stirred at 100 ° C for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (30 mL x 2), the organic phases were combined, washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product, which was separated by column chromatography (petroleum ether/ethyl acetate = 5:1) to obtain the title compound (1.44 g).

MS m/z(ESI):312.1[M+H] ⁺

Step 2: Synthesis of 4-(4-(dimethoxymethyl)piperidin-1-yl)-2-fluorobenzoic acid (O-3)

At room temperature, methyl 4-(4-(dimethoxymethyl)piperidin-1-yl)-2-fluorobenzoate (O-2, 1.44 g, 4.63 mmol) was dissolved in tetrahydrofuran (10 mL), methanol (10 mL) and water (10 mL), and lithium hydroxide monohydrate (486 mg, 11.58 mmol) was added. The mixture was reacted at room temperature for 2 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and water (20 mL) was added to dilute it. The pH was adjusted to about 3-4 with 2N hydrochloric acid aqueous solution. Solids precipitated, filtered, and the filter cake was washed with water to obtain a crude product. The crude product was dissolved in ethyl acetate (20 mL), washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound (1.30 g).

MS m/z(ESI):298.1[M+H] ⁺

Step 3: Synthesis of 4-(4-(dimethoxymethyl)piperidin-1-yl)-N-(2,6-dioxopiperidin-3-yl)-2-fluorobenzamide (O-4)

At room temperature, 4-(4-(dimethoxymethyl)piperidin-1-yl)-2-fluorobenzoic acid (O-3, 1.30 g, 4.37 mmol) and 3-aminopiperidine-2,6-dione hydrochloride (862 mg, 5.24 mmol) were dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (1.69 g, 13.11 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.99 g, 5.24 mmol) were added. The mixture was reacted at room temperature for 1 hour. After the reaction, the reaction solution was diluted with water (30 mL), extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was separated by column chromatography (petroleum ether/ethyl acetate = 1:1) to obtain the title compound (1.30 g).

MS m/z(ESI):408.2[M+H] ⁺

Step 4: Synthesis of N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(4-formylpiperidin-1-yl)benzamide (Intermediate O)

At room temperature, 4-(4-(dimethoxymethyl)piperidin-1-yl)-N-(2,6-dioxopiperidin-3-yl)-2-fluorobenzamide (O-4, 700 mg, 1.72 mmol) was dissolved in acetone (9 mL), and aqueous hydrochloric acid solution (3 mL, 2N) was added, and the temperature was raised to 80°C and stirred for 1 hour. After the reaction was completed, the reaction solution was cooled to room temperature, saturated aqueous sodium bicarbonate solution was added to adjust the pH to 7-8, and extracted with dichloromethane (50 mL x 3), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound (520 mg).

MS m/z(ESI):362.1[M+H] ⁺

Intermediate P: Synthesis of 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxylic acid

Step 1: Synthesis of tert-butyl 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxylate (P-1)

3-(6-fluoropyridin-3-yl)piperidine-2,6-dione (C-3, 200 mg, 960.67 μmol) was dissolved in dimethyl sulfoxide (1 mL), and tert-butyl piperidine-4-carboxylate (534 mg, 2.88 mmol) and N,N-diisopropylethylamine (248 mg, 1.92 mmol, 334.65 μL) were added, and microwave reaction was performed at 120 ° C for 2 h. LCMS showed that the main product was the product. Ethyl acetate (5 mL) and water were added to the reaction solution for extraction, washed with saturated brine, dried over anhydrous sodium sulfate, purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1), and concentrated under reduced pressure to remove the solvent to obtain the title compound (217 mg).

MS m/z(ESI):374.10[M+H] ⁺

Step 2: Synthesis of 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxylic acid (Intermediate P)

1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxylic acid tert-butyl ester (P-1, 171 mg, 457.90 μmol) was dissolved in a mixed solution of dichloromethane (2 mL) and trifluoroacetic acid (1 mL) and reacted at room temperature for 2 hours. The reaction was monitored by LCMS. After the reaction was completed, the solvent was removed by concentration under reduced pressure to obtain the title compound (145 mg).

MS m/z(ESI):318.10[M+H] ⁺

Intermediate Q: Synthesis of 1-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)piperidine-4-carboxaldehyde

Referring to the synthesis method of intermediate G, intermediate G-1 was replaced with 3-(4-bromo-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione, and intermediate Q was prepared in the same manner.

Intermediate R: Synthesis of 1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperidine-4-carboxaldehyde

Step 1: Synthesis of 3-((6-bromo-1-methyl-1H-indazol-3-yl)amino)propanoic acid (R-2)

To a stirred solution of 6-bromo-1-methyl-1H-indazole-3-amine (R-1, 10.00 g, 44.248 mmol) in glacial acetic acid (6.29 g, 104.87 mmol) and water (30 mL) was added acrylic acid (3.189 g, 44.248 mmol) at room temperature. The mixture was stirred at 105 ° C. under nitrogen protection for 20 h. The residue obtained by concentration was separated by silica gel column chromatography (dichloromethane/methanol=30/1) to give the title compound (4.2 g).

MS m/z(ESI):298.14[M+H] ⁺

Step 2: Synthesis of 1-(6-bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (R-3)

At room temperature, 3-((6-bromo-1-methyl-1H-indazol-3-yl)amino)propionic acid (R-2, 4.2 g, 14.14 mmol) and urea (4.24 g, 70.71 mmol) were dissolved in glacial acetic acid (50.0 mL), nitrogen was replaced, the temperature was raised to 140°C, and the reaction mixture was stirred at 140°C for 16 hours. After cooling to room temperature, concentrated hydrochloric acid (10.0 mL) was added and the reaction was heated to 140°C again and stirred for 30 minutes. After the reaction was completed, the residue was concentrated and separated by silica gel column chromatography (dichloromethane/methanol=30/1) to obtain the title compound (1.3 g).

MS m/z(ESI):323.15[M+H] ⁺

Step 3: Synthesis of 1-(6-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (R-4)

At room temperature, 1-(6-bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (R-3, 500 mg, 1.548 mmol), 4-(dimethoxymethyl)piperidine (492.26 mg, 3.096 mmol), cesium carbonate (1.51 g, 4.644 mmol) and [1,3-bis[2,6-bis(1-ethylpropyl)phenyl]-4,5-dichloro-1,3-dihydro-2H-imidazol-2-ylidene]dichloro(2-methylpyridine)palladium (78.03 mg, 0.093 mmol) were dissolved in 1,4-dioxane (10 mL), nitrogen was replaced, the temperature was raised to 100°C, and the reaction mixture was stirred at 100°C for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and dichloromethane (30 mL) was added to dilute it. AcOH (2 mL, 2N) was added dropwise to adjust the pH to 6. The solution was diluted with water and the organic phase was washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (dichloromethane/methanol = 15/1) to obtain the title compound (430 mg).

MS m/z(ESI):402.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.50(s,1H),7.45(d,J＝8.8Hz,1H),6.93-6.83(m,2H),4.10(d,J＝6.4Hz,1H),3.90-3.87(m,5 H),3.80(d,J＝12.4Hz,2H),3.28(s,6H),2.74-2.71(m,4H),1.76-1.73(m,3H),1.40-1.38(m,2H).

Step 4: Synthesis of 1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperidine-4-carbaldehyde (Intermediate R)

1-(6-(4-(dimethoxymethyl)piperidin-1-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (R-4, 30 mg, 74.73 μmol) was dissolved in tetrahydrofuran (1 mL), diluted hydrochloric acid (2M, 373.64 μL) was added, heated to 60°C, and reacted for 2 hours. LCMS monitoring showed that the reaction was complete, and the reaction solution was concentrated under reduced pressure, and an appropriate amount of water was added and freeze-dried to obtain the crude title compound (25 mg), which was directly carried out to the next step.

Intermediate S: Synthesis of 3-(5-(piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-((tetrahydro-2H-pyran-4-yl)amino)-5H-pyrido[3,2-b]indole-7-carbonitrile

Step 1: Synthesis of 5-bromo-2-chloro-3-nitro-N-(tetrahydro-2H-pyran-4-yl)pyridin-4-amine (S-1)

Dissolve 5-bromo-2,4-dichloro-3-nitropyridine (2g, 7.36mmol) in N,N-dimethylformamide (10mL), add tetrahydropyran-4-amine (744.04mg, 7.36mmol) and triethylamine (1.12g, 11.03mmol, 1.54mL), and stir at room temperature for 3 hours. Monitor the reaction by LCMS. After the reaction is completed, add the reaction solution to an appropriate amount of water, extract with ethyl acetate (20mL*3), wash the organic phase three times with saturated brine, dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify by silica gel column chromatography (petroleum ether/ethyl acetate=10/1) to obtain the title compound (2.08g).

MS m/z(ESI):336.0/338.0[M+H] ⁺

Step 2: Synthesis of 4-(5-bromo-3-nitro-4-((tetrahydro-2H-pyran-4-yl)amino)pyridin-2-yl)-3-fluorobenzonitrile (S-2)

Dissolve 5-bromo-2-chloro-3-nitro-N-(tetrahydro-2H-pyran-4-yl)pyridin-4-amine (1.8 g, 5.35 mmol) in 1,4-dioxane (20 mL), add tetrakistriphenylphosphine palladium (618.00 mg, 534.81 μmol), 3-fluoro-4-[4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl]benzonitrile (1.32 g, 5.35 mmol), stir at room temperature for 12 hours. LCMS monitoring shows that the reaction is relatively complete. Add appropriate amount of water and dichloromethane (20 mL*3) to the reaction solution for extraction, combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and obtain the residue by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain the title compound (800 mg).

MS m/z(ESI):421.0/423.0[M+H] ⁺

Step 3: Synthesis of tert-butyl 4-(5-(6-(4-cyano-2-fluorophenyl)-5-nitro-4-((tetrahydro-2H-pyran-4-yl)amino)pyridin-3-yl)-1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (S-3)

Under nitrogen atmosphere, 4-(5-bromo-3-nitro-4-((tetrahydro-2H-pyran-4-yl)amino)pyridin-2-yl)-3-fluorobenzonitrile (700 mg, 1.66 mmol) was dissolved in 1,4-dioxane (5 mL), and tert-butyl 4-(1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (449.28 mg, 1.66 mmol) and trimethylacetic acid (50. 92mg, 498.55μmol), cuprous iodide (63.30mg, 332.37μmol), palladium acetate (74.62mg, 332.37μmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (384.63mg, 664.74μmol) and cesium carbonate (1.62g, 4.99mmol), add and heat to 100℃ and stir for 12 hours. LCMS shows that the reaction is complete. The reaction solution is filtered with diatomaceous earth, and the filtrate is concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1) to obtain the title compound (120mg).

MS m/z(ESI):611.2[M+H] ⁺

Step 4: Synthesis of tert-butyl 4-(5-(5-amino-6-(4-cyano-2-fluorophenyl)-4-((tetrahydro-2H-pyran-4-yl)amino)pyridin-3-yl)-1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (S-4)

Tert-butyl 4-(5-(6-(4-cyano-2-fluorophenyl)-5-nitro-4-((tetrahydro-2H-pyran-4-yl)amino)pyridin-3-yl)-1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (120 mg, 196.51 μmol) was dissolved in tetrahydrofuran (5 mL) and ethanol (5 mL), 10% palladium carbon (55% wt H ₂ O, 24 mg) was added, and the mixture was heated to 50° C. under a hydrogen atmosphere and reacted overnight. LCMS monitoring showed that the reaction was complete. The reaction solution was filtered using diatomaceous earth, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (dichloromethane/methanol=100/1) to obtain (44 mg).

MS m/z(ESI):581.3[M+H] ⁺

Step 5: Synthesis of tert-butyl 4-(5-(7-cyano-4-((tetrahydro-2H-pyran-4-yl)amino)-5H-pyrido[3,2-b]indol-3-yl)-1,3,4-thiadiazol-2-yl)piperazine-1-carboxylate (S-5)

4-(5-(5-amino-6-(4-cyano-2-fluorophenyl)-4-((tetrahydro-2H-pyran-4-yl)amino)pyridin-3-yl)-1.3,4-thiadiazol-2-yl)piperazine-1-carboxylic acid tert-butyl ester (44 mg, 75.77 μmol) and cesium carbonate (123.44 mg, 378.87 μmol) were dissolved in N,N-dimethylformamide (1 mL), and the temperature was raised to 100°C for overnight reaction. LCMS monitoring showed that the reaction was complete, and an appropriate amount of water was added to the reaction solution, and ethyl acetate (5 mL*3) was used for extraction. The organic phase was washed three times with saturated brine, and after reduced pressure concentration, thin layer chromatography purification (dichloromethane/methanol=10/1) was obtained (3.3 mg).

MS m/z(ESI):561.3[M+H] ⁺

Step 6: Synthesis of 3-(5-(piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-((tetrahydro-2H-pyran-4-yl)amino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Intermediate S)

Dissolve tert-butyl 4-[5-[7-cyano-4-[tetrahydropyran-4-ylamino]-5H-pyrido[3,2-b]indol-3-yl]-1,3,4-thiadiazol-2-yl]piperazine-1-carboxylate (3.3 mg, 5.89 μmol) in methanol (1.99 mL) and add a 1,4-dioxane solution of hydrochloric acid (4M, 29.43 μL). React at room temperature for 3 hours. LCMS monitoring shows reaction. Filter the solid precipitated in the reaction solution and collect the solid to obtain the title compound (1.3 mg).

MS m/z(ESI):461.1[M+H] ⁺

Intermediate T: Synthesis of 3-(5-(3,8-diazabicyclo[3.2.1]octan-3-yl)-1,3,4-thiadiazol-2-yl)-4-isopropylamino-5H-pyrido[3,2-b]indole-7-carbonitrile

Step 1: Synthesis of tert-butyl 3-(1,3,4-thiadiazol-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (T-2)

Dissolve 2-bromo-1,3,4-thiadiazole (2g, 12.12mmol) in N,N-dimethylformamide (20mL), add tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (T-1, 2.83g, 13.33mmol) and N,N-diisopropylethylamine (3.13g, 24.24mmol, 4.22mL), heat to 100℃ and stir for 12 hours. LCMS monitoring shows that the reaction is complete. Add appropriate amount of water to the reaction solution, extract with ethyl acetate (30mL*3), wash the organic phase three times with saturated brine, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain a residue for silica gel column chromatography Spectral purification (petroleum ether/ethyl acetate=1/1) gave the title compound (3 g).

MS m/z(ESI):297.1[M+H] ⁺

Step 2: Synthesis of tert-butyl 3-(5-(6-(4-cyano-2-fluorophenyl)-4-isopropylamino-5-nitropyridin-3-yl)-1,3,4-thiadiazol-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (T-3)

Under nitrogen atmosphere, 4-(5-bromo-4-isopropylamino-3-nitropyridin-2-yl)-3-fluorobenzonitrile (A-3, 1.02 g, 2.69 mmol) was dissolved in 1,4-dioxane (30 mL), and tert-butyl 3-(1,3,4-thiadiazol-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (T-2, 1.2 g, 4.05 mmol) and pivalic acid ( 82.70mg, 809.75μmol), cuprous iodide (102.81mg, 539.83μmol), palladium acetate (121.20mg, 539.83μmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (624.71mg, 1.08mmol) and cesium carbonate (2.64g, 8.10mmol), add and heat to 100℃ and stir for 12 hours. LCMS monitoring shows that the reaction is complete. The reaction solution is filtered through diatomaceous earth, the filtrate is collected, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1) to obtain the title compound (700mg).

MS m/z(ESI):595.2[M+H] ⁺

Step 3: Synthesis of tert-butyl 3-(5-(5-amino-6-(4-cyano-2-fluorophenyl)-4-isopropylaminopyridin-3-yl)-1,3,4-thiadiazol-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (T-4)

3-(5-(6-(4-cyano-2-fluorophenyl)-4-isopropylamino-5-nitropyridin-3-yl)-1,3,4-thiadiazol-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (T-3, 700 mg, 1.18 mmol) was dissolved in tetrahydrofuran (5 mL) and ethanol (5 mL), 10% palladium carbon (55% wt H ₂ O, 140 mg) was added, and the mixture was heated to 50° C. under a hydrogen atmosphere for 3-5 hours. LCMS monitoring showed that the reaction solution was filtered using diatomaceous earth, the filtrate was collected and concentrated under reduced pressure, and then purified by silica gel column chromatography (dichloromethane/methanol=10/1) to obtain the title compound (505 mg).

m/z:ES ⁺ [M+H] ⁺ =565.3

Step 4: Synthesis of tert-butyl 3-(5-(7-cyano-4-isopropylamino-5H-pyrido[3,2-b]indol-3-yl)-1,3,4-thiadiazol-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (T-5)

Tert-butyl 3-(5-(5-amino-6-(4-cyano-2-fluorophenyl)-4-(isopropylamino)pyridin-3-yl)-1,3,4-thiadiazol-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (T-4, 75 mg, 132.82 μmol) and cesium carbonate (216 mg, 662.94 μmol) were dissolved in N,N-dimethylformamide (2 mL). The mixture was stirred at 100° C. for 16 hours. The filtrate was filtered and concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (dichloromethane/methanol=20/1) to give the title compound (30 mg).

MS m/z(ESI):545.20[M+H] ⁺

Step 5: Synthesis of 3-(5-(3,8-diazabicyclo[3.2.1]octan-3-yl)-1,3,4-thiadiazol-2-yl)-4-isopropylamino-5H-pyrido[3,2-b]indole-7-carbonitrile (Intermediate T)

3-(5-(7-cyano-4-isopropylamino-5H-pyrido[3,2-b]indol-3-yl)-1,3,4-thiadiazol-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (T-5, 30 mg, 55.08 μmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added thereto. The mixture was stirred at 25 ° C for 16 hours. The mixture was filtered and concentrated to dryness to give the title compound (40 mg).

MS m/z(ESI):445.10[M+H] ⁺ .

Intermediate V: Synthesis of 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)-4-hydroxypiperidine-4-carboxaldehyde

Step 1: Synthesis of 3-(6-(4-hydroxy-4-(hydroxymethyl)piperidin-1-yl)pyridin-3-yl)piperidine-2,6-dione (V-1)

3-(6-fluoropyridin-3-yl)piperidine-2,6-dione (C-3, 450 mg, 2.16 mmol) and 4-(hydroxymethyl)piperidin-4-ol (1.03 mg, 7.85 mmol) were dissolved in dimethyl sulfoxide (5 mL), N,N-diisopropylethylamine (558.70 mg, 4.32 mmol) was added, and the temperature was raised to 120°C for microwave reaction for 2.5 h. After LC-MS showed that the reaction was complete, 100 mL of ethyl acetate was added for dilution, and the organic phase was washed with water (30 mL) and saturated brine (30 mL), and dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/ethyl acetate = 1/1) to obtain the title compound (320 mg).

Step 2: Synthesis of 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)-4-hydroxypiperidine-4-carbaldehyde (Intermediate V)

At room temperature, 3-(6-(4-hydroxy-4-(hydroxymethyl)piperidin-1-yl)pyridin-3-yl)piperidine-2,6-dione (V-1, 32 mg, 0.10 mmol) was dissolved in dimethyl sulfoxide (1 mL), and 2-iodoacylbenzoic acid (56 mg, 0.20 mmol) was slowly added under stirring. The reaction was stirred at room temperature for 2 hours. After the reaction was completed, it was concentrated under reduced pressure. The residue was separated by silica gel column chromatography (dichloromethane/methanol=10/1) to give the title compound (22 mg).

Example 1: Synthesis of 3-(5-(4-((1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazole-2-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 1)

The crude product of 4-(isopropylamino)-3-(5-(piperazine-1-yl)-1,3,4-thiadiazol-2-yl)-5H-pyrido[3,2-b]indole-7-carbonitrile (Intermediate A, 15 mg) and 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxaldehyde (Intermediate C, 15 mg, 50 μmol) were dissolved in N,N-dimethylformamide (1 mL), sodium acetate (9 mg, 107 μmol) was added and reacted at room temperature for 4 h, and then sodium triacetoxyborohydride (15 mg, 71 μmol) was added and the reaction continued overnight. LC-MS detection showed that the reaction was complete. The reaction solution was dropped into a saturated sodium bicarbonate solution (4 mL), and a solid precipitated. It was filtered and the filter cake was purified by silica gel column chromatography (methanol/dichloromethane = 0-10% gradient elution) to obtain the title compound (2 mg).

MS m/z (ESI): 704.3 [M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ )δ11.41(s,1H),10.73(s,1H),8.89(d,J＝9.0Hz,1H),8.43(s,1H),8.25(d,J＝8.1Hz,1H),8.05(s,1H),7.94 (d,J＝2.5Hz,1H),7.59-7.52(m,1H),7.40-7.32(m,1H),6.78(d,J＝8.9Hz,1H),4.61-4.50(m,1H),4.26(d,J＝ 12.9Hz,2H),3.77-3.67(m,1H),3.60-3.53(m,3H),2.80(t,J＝12.3Hz,2H),2.74-2.60(m,1H),2.58-2.50(m ,5H),2.24(d,J＝6.8Hz,2H),2.23-2.09(m,1H),2.04-1.94(m,1H),1.84-1.76(m,6H),1.33(d,J＝6.2Hz,6H).

Example 2: Synthesis of 3-(5-(4-(((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 2)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate D, and compound 2 was prepared in the same manner.

MS m/z (ESI): 772.3 [M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ )δ11.44(s,1H),11.07(s,1H),8.94(d,J＝8.9Hz,1H),8.44(s,1H),8.26(d,J＝8.1Hz,1H),8.05(t,J＝1.0Hz,1H),7.66(d,J ＝8.5Hz,1H),7.58(dd,J＝8.1,1.4Hz,1H),7.32(d,J＝2.3Hz,1H),7.24(d,J＝9.0Hz,1H),5.07(dd,J＝12.9,5.4Hz,1H),4.59 -4.47(m,1H),4.06(d,J＝13.1Hz,2H),3.57(s,4H),2.97(q,J＝14.8,13.6Hz,2H),2.92-2.82(m,1H),2.60(d,J＝2.0Hz,1H) ,2.57-2.51(m,6H),2.23(d,J＝6.7Hz,2H),2.07-1.96(m,1H),1.93-1.79(m,3H),1.52-1.41(m,1H),1.33(d,J＝6.1Hz,6H).

Example 3: Synthesis of 3-(1-(1-((1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidin-4-yl)methyl)piperidin-4-yl)-1H-1,2,3-triazol-4-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 3)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and compound 3 was prepared in the same manner.

MS m/z (ESI): 686.3 [M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ )δ11.34(s,1H),10.79(s,1H),8.93(s,1H),8.66(s,1H),8.30–8.24(m,1H),8.19–8.12(m,1H),8. 02(s,1H),7.98–7.93(m,1H),7.59–7.53(m,1H),7.39–7.34(m,1H),6.82–6.77(m,1H),4.63–4.56( m,1H),4.49–4.33(m,2H),4.32–4.23(m,2H),3.77–3.67(m,2H),3.03–2.98(m,1H),2.85–2.60(m,4 H),2.26–2.08(m,7H),2.01–1.95(m,1H),1.83–1.76(m,2H),1.29–1.24(m,6H),1.19–1.05(m,4H).

Example 4: Synthesis of 3-(1-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-yl)-1H-1,2,3-triazol-4-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 4)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and intermediate C was replaced by intermediate D, and compound 4 was prepared in the same manner.

MS m/z(ESI):754.3[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ )δ11.34(s,1H),11.06(s,1H),8.93(s,1H),8.66(s,1H),8.30–8.21(m,1H),8.20–8.08(m,1H),8.06–7.9 8(m,1H),7.70–7.62(m,1H),7.59–7.51(m,1H),7.35–7.28(m,1H),7.28–7.20(m,1H),5.12–5.01(m,1H), 4.66–4.55(m,1H),4.47–4.35(m,1H),4.13–3.99(m,2H),3.05–2.91(m,4H),2.91–2.83(m,1H),2.62–2.5 4(m,2H),2.27–2.21(m,2H),2.21–1.97(m,7H),1.92–1.79(m,3H),1.31–1.24(m,6H),1.22–1.12(m,2H).

Example 5: Synthesis of 3-(1-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 5)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and intermediate C was replaced by intermediate E, and compound 5 was prepared in the same manner.

MS m/z(ESI):740.4[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.96(s,1H),10.18(s,1H),9.11(s,1H),8.90(s,1H),8.65(s,1H),8.26(s,1H),7 .74–7.65(m,1H),7.45–7.38(m,1H),7.32–7.26(m,1H),7.20(s,2H),5.12–5.07(m,1 H),4.38–4.18(m,2H),3.84–3.77(m,3H),3.08(s,2H),2.82–2.78(m,2H),2.60–2.56 (m,3H),1.99–1.93(m,3H),1.43–1.41(m,8H),1.24–1.23(m,6H),0.86–0.83(m,4H).

Example 6: Synthesis of 3-(1-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)piperidin-4-yl)methyl)piperidin-4-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 6)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and intermediate C was replaced by intermediate F, and compound 6 was prepared in the same manner.

MS m/z(ESI):740.4[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.35(s,1H),11.01-10.90(m,1H),8.95-8.86(m,1H),8.68-8.62(m,1H),8.29-8.23(m,1H),8.18 -8.07(m,1H),8.05-8.00(m,1H),7.58-7.48(m,2H),7.08-7.01(m,2H),5.08-5.00(m,1H),4.64-4.5 6(m,1H),4.46-4.36(m,1H),4.36-4.16(m,3H),3.93-3.86(m,2H),3.04-2.96(m,2H),2.96-2.76(m, 4H),2.27-2.22(m,2H),2.21-2.08(m,6H),1.85-1.80(m,2H),1.29-1.26(m,6H),1.26-1.21(m,4H).

Example 7: Synthesis of 3-(1-(1-(2-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo(d)imidazol-5-yl)piperidin-4-yl)methyl)piperidin-4-yl)-1H-1,2,3-triazol-4-yl)-4-(isopropylamino)-5H-pyrido(3,2-b)indole-7-carbonitrile (Compound 7)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and intermediate C was replaced by intermediate G, and compound 7 was prepared in the same manner.

MS m/z(ESI):755.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.34(s,1H),11.06(s,1H),8.94(s,1H),8.66(s,1H),8.29–8.22(m,1H),8.19–8.13(m,1H),8.05–8.00(m,1H),7.58 –7.53(m,1H),6.95–6.90(m,1H),6.85–6.82(m,1H),6.67–6.61(m,1H),5.32–5.26(m,1H),4.65–4.57(m,1H),4.45–4.3 9(m,1H),3.64–3.57(m,2H),3.05–2.98(m,2H),2.68–2.62(m,3H),2.28–2.24(m,2H),2.19–2.12(m,5H),2.09–2.06(m, 1H),2.02–1.96(m,2H),1.87–1.80(m,2H),1.72–1.63(m,2H),1.29–1.26(m,6H),1.25–1.23(m,3H),0.87–0.80(m,2H).

Example 8: Synthesis of 3-(1-(1-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2-azaspiro[3.5]non-7-yl)methyl)piperidin-4-yl)-1H-1,2,3-triazol-4-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 8)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and intermediate C was replaced by intermediate H, and compound 8 was prepared in the same manner.

MS m/z(ESI):794.4[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.35(s,1H),11.06(s,1H),8.94-8.89(m,1H),8.68-8.64(m,1H),8.29-8.23(m,1H),8.19-8.11(m,1H),8.03 -7.99(m,1H),7.66-7.61(m,1H),7.57-7.53(m,1H),6.78-6.74(m,1H),6.66-6.61(m,1H),5.09-5.01(m,1H),4.6 7-4.55(m,1H),4.47-4.36(m,1H),3.77-3.68(m,4H),3.06-2.84(m,3H),2.64-2.53(m,2H),2.28-2.09(m,7H),1 .95-1.88(m,2H),1.78-1.72(m,2H),1.58-1.47(m,3H),1.29-1.26(m,6H),1.25-1.21(m,2H),1.04-0.91(m,2H).

Example 9: Synthesis of 3-(1-(1-((1-(4-((2,6-dioxopiperidin-3-yl)oxy)phenyl)piperidin-4-yl)methyl)piperidin-4-yl)-1H-1,2,3-triazol-4-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 9)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and intermediate C was replaced by intermediate I, and compound 9 was prepared in the same manner.

MS m/z(ESI):701.30[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.28(s,1H),10.82(s,1H),8.86(s,1H),8.59(s,1H),8.21-8.16(m,1H),8.12-8.06(m, 1H),7.98-7.94(m,1H),7.52-7.45(m,1H),6.87-6.77(m,4H),4.97-4.90(m,1H),4.58-4.4 8(m,1H),4.40-4.29(m,1H),3.50-3.42(m,2H),2.98-2.89(m,2H),2.66-2.46(m,5H),2.21 -2.15(m,2H),2.14-2.00(m,8H),1.80-1.70(m,2H),1.22-1.19(m,6H),1.18-1.10(m,2H).

Example 10: Synthesis of 3-(5-(4-((1-(5-(2,4-dioxohexahydropyrimidin-1-yl)-pyridin-2-yl)-piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazole-2-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 10)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate J (prepared according to patent WO2023023255A1), and compound 10 was prepared in the same way.

MS m/z(ESI):705.2[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.53–11.40(m,1H),10.34(s,1H),8.97–8.90(m,1H),8.43(s,1H),8.29–8.22(m,1H),8.0 9–8.00(m,2H),7.60–7.53(m,1H),7.50–7.44(m,1H),6.87–6.80(m,1H),4.60–4.51(m,1H),4. 32–4.24(m,2H),3.71–3.66(m,2H),3.62–3.47(m,4H),2.86–2.77(m,2H),2.73–2.66(m,3H),2 .26–2.19(m,2H),1.87–1.72(m,4H),1.36–1.31(m,6H),1.11–1.05(m,2H),0.79–0.73(m,2H).

Example 11: Synthesis of 3-(5-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazole-2-yl)-4-(isopropylamino)-5H-pyrido(3,2-b)indole-7-carbonitrile (Compound 11)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate K (prepared according to patent WO2022099117A1), and compound 11 was prepared in the same way.

MS m/z(ESI):772.3(M+H) ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.45(s,1H),11.10(s,1H),8.98–8.92(m,1H),8.44(s,1H),8.28–8.23(m,1H),8.05(s,1H),7.72–7.6 5(m,1H),7.60–7.55(m,1H),7.36–7.31(m,2H),5.14–5.05(m,1H),4.59–4.51(m,1H),3.74–3.68(m,2H), 3.58–3.54(m,3H),2.92–2.87(m,2H),2.57–2.53(m,4H),2.30–2.26(m,2H),2.06–1.99(m,1H),1.88–1.8 3(m,2H),1.40–1.39(m,1H),1.34–1.32(m,6H),1.24–1.22(m,2H),0.97–0.93(m,1H),0.86–0.82(m,3H).

Example 12: Synthesis of 3-(5-(4-((1-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-isopropylamino-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 12)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate G, and compound 12 was prepared in the same manner.

MS m/z(ESI):773.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.56(s,1H),11.09(s,1H),9.02(s,1H),8.49(s,1H),8.33–8.25(m,1H),8.08(s,1H),7.65–7.57 (m,1H),6.99–6.92(m,1H),6.86(s,1H),6.70–6.63(m,1H),5.35–5.25(m,2H),4.64–4.51(m,1H),4. 12(s,1H),3.75–3.58(m,6H),3.33–3.30(m,5H),2.96–2.83(m,2H),2.72–2.68(m,3H),2.65–2.59(m ,2H),2.22–2.14(m,1H),2.04–1.96(m,2H),1.89–1.82(m,2H),1.37–1.32(m,6H),1.25–1.21(m,1H).

Example 13: Synthesis of 3-(5-(4-((1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazole-2-yl)-4-isopropylaminobenzofurano[3,2-b]pyridine-7-carbonitrile (Compound 13)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate L, and compound 13 was prepared in the same manner.

MS m/z(ESI):705.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.95(s,1H),9.35(s,1H),8.69-8.61(m,1H),8.54-8.48(m,1H),8.45-8.36 (m,1H),7.97-7.93(m,1H),7.35-7.32(m,1H),7.11-7.06(m,1H),4.84-4.71(m, 1H),4.40-4.28(m,2H),4.13-4.07(m,2H),3.99-3.88(m,4H),2.76-2.62(m,2H ),2.23-2.14(m,3H),2.04-1.98(m,10H),1.42-1.40(m,6H),1.36-1.35(m,2H).

Example 14: 3-(5-(4-((1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-6-yl)piperidin-4-yl)methyl)piperazine-1- Synthesis of 1,3,4-thiadiazol-2-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 14)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate M, and compound 14 was prepared in the same manner.

MS m/z(ESI):757.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.62–11.40(m,1H),10.86(s,1H),8.97–8.87(m,1H),8.40(s,1H),8.30–8.19(m,1H),8 .04(s,1H),7.53–7.46(m,2H),6.94–6.89(m,1H),6.86–6.82(m,1H),4.78–4.68(m,1H),4. 28–4.24(m,1H),3.89(s,3H),3.83–3.77(m,2H),3.61–3.54(m,4H),2.80–2.60(m,5H),2.5 9–2.53(m,4H),2.36–2.23(m,3H),2.20–2.13(m,1H),1.91–1.72(m,4H),1.36–1.30(m,6H).

Example 15: Synthesis of 3-(5-(4-((1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-isopropylamino-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 15)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate N, and compound 15 was prepared in the same manner.

MS m/z(ESI):757.2[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.46(s,1H),10.88(s,1H),9.00–8.86(m,1H),8.44(s,1H),8.30–8.21(m,1H),8.05 (s,1H),7.62–7.53(m,1H),7.47–7.32(m,2H),7.09–6.94(m,3H),4.59–4.49(m,1H),4.3 6–4.31(m,1H),4.28–4.22(m,4H),3.67–3.53(m,4H),2.79–2.55(m,3H),2.32(s,3H),2 .24–2.13(m,2H),2.08(s,3H),1.94–1.86(m,2H),1.41–1.30(m,6H),0.93–0.81(m,3H).

Example 16: Synthesis of 3-(5-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-isopropylamino-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 16)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate E, and compound 16 was prepared in the same manner.

MS m/z(ESI):758.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.65–11.28(m,1H),10.97(s,1H),8.98–8.87(m,1H),8.41(s,1H),8.28–8.19(m,1H),8.04(s,1H),7.57–7. 48(m,1H),7.44–7.39(m,1H),7.28–7.24(m,1H),7.18–7.14(m,1H),5.12–5.06(m,1H),4.74–4.59(m,1H),4.37– 4.17(m,3H),3.81–3.72(m,2H),3.59–3.53(m,3H),3.18–3.15(m,1H),2.97–2.85(m,2H),2.77–2.69(m,2H),2. 42–2.30(m,2H),2.30–2.20(m,2H),2.03–1.95(m,2H),1.87–1.72(m,4H),1.51–1.41(m,2H),1.35–1.30(m,6H).

Example 17: Synthesis of 3-(5-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 17)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate O, and compound 17 was prepared in the same manner.

MS m/z(ESI):764.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.43(s,1H),10.83(s,1H),8.97-8.90(m,1H),8.47-8.42(m,1H), 8.28-8.23(m,1H),8.07-8.03(m,1H),8.03-7.95(m,1H),7.67-7.54(m ,2H),6.85-6.69(m,2H),4.78-4.68(m,1H),4.58-4.46(m,1H),3.92- 3.85(m,2H),3.62-3.52(m,4H),2.90-2.81(m,2H),2.58-2.52(m,3H), 2.28-2.20(m,2H),2.21-1.96(m,3H),1.87-1.75(m,3H),1.38-1.29(m,6H),1.29-1.07(m,4H).

Example 18: Synthesis of 3-(5-(4-(1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carbonyl)piperazine-1-yl)-1,3,4-thiadiazol-2-yl)-4-isopropylamino-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 18)

4-(Isopropylamino)-3-(5-(piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-5H-pyrido[3,2-b]indole-7-carbonitrile (Intermediate A, 153 mg, 365.54 μmol) was dissolved in N,N-dimethylformamide (2 mL), 1-methylimidazole (75 mg, 913.85 μmol) was added to make it alkaline, and N,N,N',N' was added. -Tetramethylchloroformamidine hexafluorophosphate (215 mg, 767.63 μmol), after stirring at room temperature for 1 hour, 1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidine-4-carboxylic acid (Intermediate P, 116 mg, 365.57 μmol) was dissolved in N,N-dimethylformamide and then adjusted to alkaline with 1-methylimidazole, reacted at room temperature for 1 hour, and the reaction was monitored by LCMS. The title compound (92 mg) was obtained by preparative liquid chromatography (YMC C18, 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 0.5% ammonium bicarbonate) and acetonitrile (acetonitrile content 50%-80%) as eluent).

MS m/z(ESI):718.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.46(s,1H),10.80(s,1H),9.01-8.90(m,1H),8.49-8.41(m,1H),8.29-8.23(m,1H),8.08-8.02(m,1H), 7.99-7.92(m,1H),7.61-7.54(m,1H),7.42-7.35(m,1H),6.86-6.79(m,1H),4.61-4.49(m,1H),4.35-4.25(m ,2H),3.83-3.71(m,3H),3.70-3.59(m,4H),3.59-3.52(m,2H),3.02-2.85(m,3H),2.73-2.62(m,1H),2.24- 2.12(m,1H),2.03-1.95(m,1H),1.77-1.69(m,2H),1.63-1.55(m,2H),1.38-1.31(m,6H),1.28-1.22(m,1H).

Example 19: Synthesis of 3-(5-(4-((1-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-isopropylamino-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 19)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate Q, and compound 19 was prepared in the same manner.

MS m/z(ESI):773.2[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.46(s,1H),11.11(s,1H),8.98-8.92(m,1H),8.49-8.41(m,1H),8.30-8.22(m,1H),8.09-8.02(m,1H), 7.62-7.55(m,1H),7.03-6.95(m,1H),6.94-6.82(m,2H),5.41-5.31(m,1H),4.59-4.51(m,1H),3.66-3.54(m ,5H),3.18-3.09(m,2H),2.98-2.80(m,2H),2.77-2.63(m,3H),2.59-2.53(m,2H),2.34-2.27(m,2H),2.09- 1.95(m,2H),1.90-1.81(m,2H),1.76-1.63(m,2H),1.38-1.28(m,6H),1.28-1.20(m,3H),0.87-0.81(m,1H).

Example 20: Synthesis of 3-(5-(4-((1-(3-(2,4-dioxohexahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-isopropylamino-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 20)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate R, and compound 20 was prepared in the same manner.

MS m/z(ESI):758.2[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.44(s,1H),10.49(s,1H),8.97–8.88(m,1H),8.44(s,1H),8.30–8 .19(m,1H),8.05(s,1H),7.61–7.53(m,1H),7.49–7.38(m,1H),6.96–6 .87(m,1H),6.85–6.77(m,1H),4.61–4.49(m,1H),3.94–3.86(m,4H),3 .84–3.79(m,2H),3.62–3.53(m,3H),2.84–2.65(m,4H),2.58–2.51(m, 5H),2.35–2.20(m,2H),2.14–1.97(m,1H),1.90–1.72(m,3H),1.36–1.29(m,5H),1.28–1.20(m,2H),0.89–0.76(m,1H).

Example 21: Synthesis of 3-(5-(4-((1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-((tetrahydro-2H-pyran-4-yl)amino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 21)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate S, and compound 21 was prepared in the same manner.

MS m/z(ESI):746.2[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.81(s,1H),9.13–8.84(m,1H),8.45(s,1H),8.32–8.21(m,1H),8.06(s,1H),8.00–7.90(m,1H) ,7.54(s,1H),7.40–7.30(m,1H),6.85–6.74(m,1H),6.05–5.91(m,1H),5.37–5.24(m,1H),5.04–4.9 3(m,1H),4.38–4.18(m,2H),3.89–3.68(m,2H),3.65–3.48(m,4H),2.86–2.63(m,4H),2.38–1.93(m, 7H),1.91–1.72(m,3H),1.60–1.43(m,2H),1.32–1.20(m,4H),1.18–1.07(m,2H),0.91–0.78(m,1H).

Example 22: Synthesis of 3-(5-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-((tetrahydro-2H-pyran-4-yl)amino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 22)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate S, and intermediate C was replaced by intermediate E, and compound 22 was prepared in the same manner.

MS m/z(ESI):800.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.55(s,1H),10.98(s,1H),9.16–8.83(m,1H),8.45(s,1H),8.35–8.20(m,1H),8.06(s,1H),7.61–7 .52(m,1H),7.46–7.38(m,1H),7.30–7.23(m,1H),7.19–7.12(m,1H),5.40–4.96(m,2H),4.57(s,1H),4. 46–4.19(m,2H),3.94–3.73(m,4H),3.65–3.52(m,5H),2.96–2.86(m,1H),2.79–2.65(m,3H),2.64–2.54 (m,4H),2.27–2.23(m,1H),2.09–1.97(m,3H),1.89–1.73(m,3H),1.57–1.45(m,2H),1.29–1.20(m,4H).

Example 23: Synthesis of 3-(5-(8-((1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)piperidin-4-yl)methyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-1,3,4-thiadiazol-2-yl)-4-isopropylamino-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 23)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate T, and compound 23 was prepared in the same manner.

MS m/z(ESI):731.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ12.04(s,1H),10.87(s,1H),8.71(s,1H),8.43–8.42(m,1H),8.20(s,1H),7.95–7.9 4(m,1H),7.72–7.69(m,1H),7.61–7.60(m,1H),7.56–7.53(m,1H),7.10–7.08(m,1H), 4.72–4.64(m,1H),4.32–4.25(m,4H),3.95–3.80(m,5H),3.01–2.95(m,4H),2.69–2.5 6(m,2H),2.28–2.16(m,3H),2.03–1.91(m,5H),1.42–1.40(m,6H),1.35–1.26(m,3H).

Example 24: Synthesis of 3-(1-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)methyl)piperidin-4-yl)-1H-1,2,3-triazol-4-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 24)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and intermediate C was replaced by intermediate K, and compound 24 was prepared in the same manner.

MS m/z(ESI):754.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.36(s,1H),11.09(s,1H),8.94(s,1H),8.66(s,1H),8.25(d,J＝8.1Hz,1H),8.15(d,J＝9.4Hz,1H ),8.03(s,1H),7.71-7.66(m,1H),7.58-7.54(m,1H),7.37-7.31(m,2H),5.14-5.06(m,1H),4.60(s,1 H),4.47-4.36(m,1H),3.72(d,J＝11.7Hz,2H),3.01(d,J＝9.4Hz,2H),2.94-2.81(m,4H),2.28(d,J＝7. 0Hz,2H),2.23-1.98(m,8H),1.85(d,J＝12.6Hz,2H),1.76(s,1H),1.27(d,J＝6.2Hz,6H),1.23(s,2H).

Example 25: Synthesis of 3-(1-(1-((1-(5-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)pyridin-2-yl)piperidin-4-yl)methyl)piperidin-4-yl)-1H-1,2,3-triazol-4-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 25)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and intermediate C was replaced by intermediate J, and compound 25 was prepared in the same manner.

MS m/z(ESI):687.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.35(s,1H),10.33(s,1H),8.92(s,1H),8.67(s,1H),8.26(d,J＝8.0Hz,1H),8.04(d ,J＝8.3Hz,2H),7.57(d,J＝8.0Hz,1H),7.48(d,J＝9.2Hz,1H),7.00-6.80(m,1H),4.52-4. 37(m,1H),4.30(s,2H),3.81-3.58(m,2H),3.10-2.94(m,2H),2.92-2.76(m,3H),2.78-2 .64(m,3H),2.40-2.29(m,2H),2.29-2.02(m,7H),1.91-1.71(m,4H),1.29-1.25(m,6H).

Example 26: Synthesis of 3-(1-(1-((1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)piperidin-4-yl)methyl)piperidin-4-yl)-1H-1,2,3-triazol-4-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 26)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate B, and intermediate C was replaced by intermediate U, and compound 26 was prepared in the same manner.

MS m/z(ESI):686.2[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.38(s,1H),10.25(s,1H),8.93(s,1H),8.66(s,1H),8.26(d,J=8.1Hz,1H),8.15(d, J＝9.4Hz,1H),8.03(t,J＝1.0Hz,1H),7.56(dd,J＝8.1,1.4Hz,1H),7.17–7.09(m,2H),6.97 –6.90(m,2H),4.60(s,1H),4.50–4.35(m,1H),3.77–3.64(m,5H),3.03-3.01(m,2H),2.6 8(t,J＝5.2Hz,4H),2.21(d,J＝38.1Hz,9H),1.82(d,J＝12.7Hz,3H),1.27(d,J＝6.2Hz,6H).

Example 27: Synthesis of 3-(5-(4-((1-(5-(2,6-dioxopiperidin-3-yl)pyridin-2-yl)-4-hydroxypiperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-(isopropylamino)-5H-pyrido[3,2-b]indole-7-carbonitrile (Compound 27)

Referring to the synthesis method of Example 1, intermediate C was replaced by intermediate V, and compound 27 was prepared in the same manner.

MS m/z(ESI):720.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.52(s,1H),10.82(s,1H),9.04–8.78(m,1H),8.46–8.23(m,1H),8.12–7.92(m,1H),7.70–7.03(m,3H),6.86–6.63(m,1H),5.43–5.2 0(m,1H),4.65–4.27(m,2H),3.96–3.67(m,2H),3.51(s,6H),2.82–2.62(m,3H),2.44–1.84(m,7H),1.68–1.29(m,8H),0.98–0.75(m,3H).

Example 28: Synthesis of 3-(5-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-(isopropylamino)benzofurano[3,2-b]pyridine-7-carbonitrile (Compound 28)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate L, and intermediate C was replaced by intermediate E, and compound 28 was prepared in the same manner.

MS m/z(ESI):379.8[(M+2H)/2] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.97(s,1H),9.19–8.97(m,1H),8.51(s,1H),8.41(s,1H),8.31–8.16(m,1H),7.95–7.75(m,1H),7.49 –7.39(m,1H),7.33–7.22(m,1H),7.22–7.12(m,1H),5.20–5.01(m,1H),4.82–4.63(m,1H),4.39–4.29(m,1 H),4.28–4.14(m,1H),3.83–3.70(m,2H),3.67–3.52(m,3H),3.01–2.81(m,2H),2.81–2.64(m,3H),2.46– 2.30(m,2H),2.29–2.13(dt,J＝25.2,7.7Hz,2H),2.04–1.69(m,5H),1.49–1.11(m,9H),0.94–0.76(m,1H).

Example 29: Synthesis of 3-(5-(4-((1-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-5)-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-(isopropylamino)benzofurano[3,2-b]pyridine-7-carbonitrile (Compound 29)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate L, and intermediate C was replaced by intermediate G, and compound 29 was prepared in the same manner.

MS m/z(ESI):774.60[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.05(s,1H),9.12–8.96(m,1H),8.53(s,1H),8.43(s,1H),8.29–8.23(m,1H),7.92–7.85 (m,1H),6.95–6.89(m,1H),6.85–6.79(m,1H),6.66–6.59(m,1H),5.31–5.24(m,1H),4.77–4 .68(m,1H),3.67–3.46(m,6H),2.97–2.80(m,2H),2.74–2.53(m,8H),2.31–2.22(m,2H),2.0 6–1.93(m,2H),1.87–1.79(m,2H),1.76–1.60(m,2H),1.44–1.33(m,6H),1.31–1.25(m,2H).

Example 30: Synthesis of 3-(5-(4-((1-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4)-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-(isopropylamino)benzofurano[3,2-b]pyridine-7-carbonitrile (Compound 30)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate L, and intermediate C was replaced by intermediate Q, and compound 30 was prepared in the same manner.

MS m/z(ESI):774.40[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ11.10(s,1H),9.11–9.01(m,1H),8.53(s,1H),8.44(s,1H),8.32–8.21(m,1H),7.94–7.83(m,1H),7.04–6.83(m,3H),5.39–5.31(m, 1H),4.78–4.67(m,1H),3.69–3.52(m,6H),3.18–3.07(m,2H),2.95–2.83(m,2H),2.75–2.54(m,8H),2.35–2.26(m,2H),2.04–1.93(m, 2H),1.90–1.80(m,2H),1.77–1.59(m,2H),1.43–1.28(m,6H).

Example 31: Synthesis of 3-(5-(4-((1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-6-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-(isopropylamino)benzofurano[3,2-b]pyridine-7-carbonitrile (Compound 31)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate L, and intermediate C was replaced by intermediate M, and compound 31 was prepared in the same manner.

MS m/z(ESI):379.3[(M+2H)/2] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.85(s,1H),9.09–9.03(m,1H),8.58–8.51(m,1H),8.47–8.40(m,1H),8.30–8.23(m,1H),7.92–7. 86(m,1H),7.51–7.43(m,1H),6.95–6.89(m,1H),6.87–6.81(m,1H),4.82–4.64(m,2H),4.29–4.22(m, 1H),3.94–3.85(m,3H),3.85–3.76(m,2H),3.58(s,3H),2.80–2.70(m,2H),2.66–2.58(m,2H),2.37–2 .22(m,3H),2.20–2.11(m,1H),1.91–1.69(m,4H),1.47–1.35(m,6H),1.23(s,3H),0.89–0.76(m,2H).

Example 32: Synthesis of 3-(5-(4-((1-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-(isopropylamino)benzofurano[3,2-b]pyridine-7-carbonitrile (Compound 32)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate L, and intermediate C was replaced by intermediate N, and compound 32 was prepared in the same manner.

MS m/z(ESI):379.3[(M+2H)/2] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.87(s,1H),9.08–9.01(m,1H),8.54–8.50(m,1H),8.43–8.40(m,1H),8.28–8.22(m, 1H),7.91–7.84(m,1H),7.41–7.34(m,1H),7.03–6.99(m,2H),4.79–4.66(m,1H),4.36–4 .30(m,1H),4.27–4.23(m,3H),3.64–3.52(m,4H),3.26(s,2H),2.75–2.61(m,5H),2.60– 2.53(m,4H),2.37–2.27(m,3H),2.22–2.11(m,2H),1.96–1.85(m,3H),1.41–1.35(m,6H).

Example 33: Synthesis of 3-(5-(4-((1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperidin-4-yl)methyl)piperazin-1-yl)-1,3,4-thiadiazol-2-yl)-4-(isopropylamino)benzofurano[3,2-b]pyridine-7-carbonitrile (Compound 33)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate L, and intermediate C was replaced by intermediate R, and compound 33 was prepared in the same manner.

MS m/z(ESI):759.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.49(s,1H),9.06(d,J＝8.4Hz,1H),8.54(s,1H),8.43(s,1H),8.27(d,J＝8.0Hz,1 H),7.89(dd,J＝8.0,1.3Hz,1H),7.44(d,J＝9.0Hz,1H),6.94–6.78(m,2H),4.79–4.69 (m,1H),3.93–3.77(m,6H),3.58(s,3H),2.80–2.69(m,3H),2.55(m,3H),2.26(m,1H) ,1.85(d,J＝13.2Hz,2H),1.38(d,J＝6.4Hz,4H),1.31–1.26(m,6H),1.25–1.24(m,4H).

Example 34: Synthesis of 4-(4-((4-(5-(7-cyano-4-(isopropylamino)benzofurano[3,2-b]pyridin-3-yl)-1,3,4-thiadiazol-2-yl)piperazin-1-yl)methyl)piperidin-1-yl)-N-(2,6-dioxopiperidin-3-yl)-2-fluorobenzamide (Compound 34)

Referring to the synthesis method of Example 1, intermediate A was replaced by intermediate L, and intermediate C was replaced by intermediate O, and compound 34 was prepared in the same manner.

MS m/z(ESI):765.3[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.83(s,1H),9.62(s,1H),9.05(d,J＝8.5Hz,1H),8.54(s,1H),8.43(s,1H),8.27(d,J＝8.0Hz,1H),8.0 0(d,J＝7.3Hz,1H),7.89(d,J＝8.0Hz,1H),7.62(s,1H),6.81(d,J＝8.9Hz,1H),4.73(d,J＝6.8Hz,1H),3.90 (d,J＝12.4Hz,3H),3.79(d,J＝13.5Hz,1H),3.57(s,3H),3.02(t,J＝12.1Hz,2H),2.88–2.75(m,4H),2.23( d,J＝6.7Hz,3H),2.18–1.96(m,4H),1.90(d,J＝13.3Hz,2H),1.80(d,J＝12.4Hz,2H),1.38(d,J＝6.4Hz,6H).

Biological activity and related properties test examples

Test Example 1 HiBiT method to detect the degradation effect of compounds on IRAK4 in THP-1-HiBiT-IRAK4 overexpressing cells

THP-1-HiBiT-IRAK4 cells (constructed by Shanghai Medicilon) were inoculated in a 96-well flat-bottom white plate (Corning, Catalog No.: 3917) with an inoculation density of 60,000 cells/25 μL/well. Then 25 μL/well of concentration gradient compounds (final concentration of the compound was: starting concentration 5 μM, 5-fold gradient dilution, a total of 8 concentrations) were added, mixed and incubated in a 37°C, 5% CO ₂ incubator for 24 h. The experimental settings were as follows: sample treatment group (cells + concentration gradient compounds), solvent control group (cells + solvent control), blank control group (only culture medium was added) 50 μL Nano-Glo HiBiT Lytic Detection buffer (Promega, Catalog No.: N3040) was added and incubated at room temperature (350 rpm) for 10 min, and the fluorescence value RLU was read using an enzyme reader (PE Envision). The degradation rate of the compound on IRAK4 % = (1-(RLU value of the sample treatment group-RLU value of the blank control group)/(RLU value of the solvent control group-RLU value of the blank control group)) × 100%, the degradation rate of the compound at each concentration and the maximum degradation rate _Dmax were calculated. The nonlinear fitting of the compound concentration-degradation rate was performed using Graphpad Prism 9 software to obtain the degradation activity curve of the compound, and the half-degradation concentration ( _DC50 ) of the compound was calculated. The larger the _Dmax value and the smaller the _DC50 value, the stronger the degradation activity of the compound. The _Dmax and _DC50 values of the tested compounds are shown in Table 1.

Table 1. D _max and DC ₅₀ of IRAK4 degraders

Test Example 2 Determination of Metabolic Stability of the Disclosed Compounds in Liver Microsomes

1. Test materials and instruments

1. Source of liver microsomes: human liver microsomes (Corning 452161), CD-1 mouse liver microsomes (Corning 452701)

2.Na ₂ HPO ₄ (Shanghai Test 20040618)

3. KH ₂ PO ₄ (Shanghai Test No. 10017608)

4.NADPH (Roche diagnostics gmbh 10107824001)

5. Tolbutamide (Sigma-aldrich T0891-100G)

6. Positive control compound Verapamil (Aladdin V303890-100mg)

7.AB5500+ LC-MS

2. Test steps

1. Preparation of 100 mM phosphate buffer (PBS): Weigh 7.098 g Na ₂ HPO ₄ , add 500 mL pure water and ultrasonically dissolve, as solution A. Weigh 3.400 g KH ₂ PO ₄ , add 250 mL pure water and ultrasonically dissolve, as solution B. Place solution A on a stirrer and slowly add solution B until the pH reaches 7.4 to prepare 100 mM PBS buffer.

2. Preparation of reaction system

Prepare the reaction system according to the table below

Table 2 Reaction system preparation information

3. Pre-incubate the reaction system in a 37°C water bath for 10 minutes. Add 40 μL 10 mM NADPH solution (NADPH is dissolved in 100 mM phosphate buffer) to the reaction system, and the final concentration of NADPH is 1 mM. Use 40 μL phosphate buffer instead of NADPH solution as a negative control. The role of the negative control is to exclude the influence of the chemical stability of the compound itself.

4. Add 4 μL 100 μM of the disclosed compound and the positive control compound verapamil to the reaction system to start the reaction. The final concentration of the compound is 1 μM.

5. After vortexing and mixing thoroughly at 0.5, 15, 30, 45 and 60 minutes, take out 50 μL of the incubation sample and terminate the reaction with 4 times of ice acetonitrile containing internal standard (50 ng/mL tolbutamide). Centrifuge the sample at 4000 rpm for 15 minutes. After centrifugation, transfer 50 μL of the supernatant to the injection plate, add 150 μL of ultrapure water and mix well for LC-MS/MS analysis.

All data were calculated using Microsoft Excel. Peak areas were detected by extracting ion spectra. The in vitro half-life (t _1/2 ) of the parent drug was detected by linear fitting the natural logarithm of the parent drug elimination percentage with time.

The in vitro half-life (t _1/2 ) was calculated by the slope (k):

in vitro t _1/2 =0.693/k

The t _1/2 values calculated by the above formula are shown in Table 3.

Table 3 Half-life values of the disclosed compounds in liver microsomes

Test Example 3 Determination of the Potential Inhibitory Effect of the Disclosed Compounds on Voltage-Gated Potassium Ion Channel hERG

1. Materials and Reagents

1 Experimental Materials and Instruments

1.1 Materials and Instruments

2 Cell lines and culture

HEK293 cell line (Cat. No. K1236) stably expressing hERG ion channel was purchased from Invitrogen. The cell line was cultured in a medium containing 85% DMEM, 10% dialyzed fetal bovine serum, 0.1mM MEM non-essential amino acid solution, 100U/mL penicillin-streptomycin solution, 25mM HEPES, 5μg/mL blasticidin and 400μg/mL geneticin. When the cell density grew to 40% to 80% of the bottom area of the culture dish, trypsin was used for digestion and passage, and passage was performed three times a week. Before the experiment, the cells were cultured in a 6cm culture dish at a density of 5×10 ⁵ , 1μg/mL doxycycline was added for induction for 48 hours, and then the cells were digested and inoculated on glass slides for subsequent manual patch clamp experiments.

3 Preparation of test compounds

1) The test compound was dissolved in DMSO and prepared into a stock solution with a final concentration of 10 mM.

2) The stock solution was diluted with DMSO at a gradient ratio of 1:3 to form three other intermediate concentration solutions, with concentrations of 3.33 mM, 1.11 mM and 0.37 mM, respectively.

3) Before the experiment, the stock solution and the intermediate concentration solution of the test compound were diluted 1000 times with NMDG extracellular solution to obtain a series of working solutions with concentrations of 10 μM, 3.33 μM, 1.11 μM and 0.37 μM. At the same time, the 10 mM stock solution was diluted 333.33 times with NMDG extracellular solution to obtain a 30 μM working solution. The content of DMSO in the working solution was 0.1-0.3% (volume ratio).

(Note: The DMSO content in the working solution should be controlled within 1% (volume ratio) to avoid cytotoxicity.)

4) After the working solution is prepared, observe with the naked eye whether there is precipitation or turbidity in the working solution. If there is, it may be due to the poor solubility of the compound in physiological solution. It can be further ultrasonicated in a water bath for 30 minutes to improve the clarity of the solution.

5) The potential inhibitory effect of the test substance on hERG channel at five concentrations of 30 μM, 10 μM, 3.33 μM, 1.11 μM and 0.37 μM was determined, and the dose-effect curve was fitted and the IC ₅₀ was calculated.

2. Experimental Methods

1) Place the small glass slide containing HEK293 cells in the culture dish into the perfusion tank of the microscope operation table.

2) Place the appropriate cell in the center of the field of view under an Olympus IX51, IX71 or IX73 inverted microscope, use the ×10 objective to find the tip of the glass electrode and place it in the center of the field of view. Then use the micromanipulator to move the electrode down while adjusting the coarse focusing knob to slowly bring the electrode closer to the cell.

3) When approaching the cell, switch to a ×40 objective lens for observation and use the micromanipulator to fine-tune the gear to gradually bring the electrode closer to the cell surface.

4) Apply negative pressure to form a seal with a resistance higher than 1 GΩ between the electrode tip and the cell membrane.

5) Compensate the instantaneous capacitive current C _fast in voltage clamp mode. Then repeatedly apply short negative pressure to break the membrane, and finally form a whole-cell recording mode.

6) Under the condition that the membrane potential is clamped at -60 mV, the slow capacitive current C _slow , cell membrane capacitance (C _m ) and input membrane resistance (R _a ) are compensated respectively.

7) After the cells are stable, the clamping voltage is changed to -90 mV, the sampling frequency is set to 20 kHz, and the filtering frequency is 10 kHz. The detection conditions for leakage current are: the clamping voltage is changed to -80 mV, and the time course is 500 ms.

8) The hERG current test method is as follows: apply a 4.8-second depolarization command voltage to depolarize the membrane potential from -80mV to +30mV, then instantly apply a 5.2-second repolarization voltage to reduce the membrane potential to -50mV to remove channel inactivation, so that the hERG tail current can be observed. The peak value of the tail current is the magnitude of the hERG current.

9) The hERG current used to detect the test compound was recorded for 120 seconds before administration to evaluate the stability of the hERG current generated by the test cells. Only stable cells within the acceptable range of the evaluation criteria can enter the subsequent compound testing.

10) Determine the inhibitory effect of the test compound on hERG current: First, the hERG current measured in the extracellular fluid containing 0.1% DMSO is used as the detection baseline. After the hERG current remains stable for at least 5 minutes, the solution containing the test compound is perfused around the cells in sequence from low concentration to high concentration. After each perfusion, wait for about 5 minutes to allow the compound to fully act on the cells and record the hERG current synchronously. After the recorded current tends to stabilize, record the last 5 hERG current values, and take the average value as the final current value at a specific concentration. After testing the compound, add 150nM dofelide to the same cell to completely inhibit its current as a positive control for the cell. At the same time, the positive compound dofelide is synchronously detected using the same patch clamp system before and after the test compound experiment to ensure the reliability and sensitivity of the entire detection system.

3. Data Analysis

The data were exported by PatchMaster software and analyzed according to the following steps:.

1) After perfusing blank solvent or 30 μM, 10 μM, 3.33 μM, 1.11 μM and 0.37 μM compound working solutions, the average values of the 5 stable continuous current values were calculated and used as the "tail current size _blank " and "tail current size _compound " respectively.

2) The current suppression percentage is calculated using the following formula.

3) The dose-effect curve was fitted using Graphpad Prism 8.0 software and the _IC50 value was calculated.

The inhibition of hERG by the disclosed compounds is shown in Table 4 below. The test data show that the hERG inhibition activity of the disclosed compounds is poor and the potential safety risk in the future is relatively small.

Table 4 Inhibitory activity of the disclosed compounds on voltage-gated potassium channel hERG

Test Example 4 Determination of Plasma Protein Binding Rate of the Disclosed Compounds

1. Test materials and instruments

1. CD-1 mouse plasma (BioIVT)

2. Human plasma (from healthy Asian volunteers from Weifang High-tech Industrial Development Zone People's Hospital (collected in the hospital with ethical approval))

3.Na ₂ HPO ₄ (Sigma S5136-500G)

4.NaH ₂ PO ₄ (Sigma S3139-500G)

5.NaCl (Sigma S5886-IKG)

6. Positive control compound ketoconazole (Sigma Lot #SLBB5223V)

7. Alprazolam (Ceriliant FE12172003)

8. Labetalol (NIFDC 100484-201001)

9. Ketoprofen (NIFDC 100337-201104)

10. Polycarbonate centrifuge tube (BECKMAN, 343778)

11. Ultracentrifuge rotor (BECKMAN, MLA-150)

12. Optima TM MAX-XP Ultracentrifuge (BECKMAN)

2. Test steps

1. Prepare DMSO working solutions of test compounds and positive control compounds at a concentration of 1 mM.

2. The water bath, incubator and ultracentrifuge are all set to 37°C, and the rotor is pre-incubated in the 37°C incubator for about 30 minutes.

3. Thaw frozen plasma (stored at -80°C) in a room temperature water bath, centrifuge at 3,220 g for 10 minutes to remove clots, and collect the supernatant into a new tube. Check and record the pH of the plasma.

4. Pre-incubate the plasma in a 37°C water bath for 5 minutes.

5. Add 3 μL of 1 mM working solution to 2997 μL of preheated plasma to obtain a spiked plasma sample with a final concentration of 1 μM.

6. Transfer 1 mL of spiked plasma sample to two ultracentrifuge tubes (n=2). Balance the ultracentrifuge tubes in a preheated rotor and incubate at 37°C, 5% _CO2 for 30 minutes. After incubation, centrifuge the ultracentrifuge tubes at 800,000 g for 3 hours at 37°C.

7. Transfer 50 μL of spiked plasma samples to 6 0.6 mL tubes respectively. The samples were incubated at 37 ° C, 5% CO ₂ for 0, 0.5 and 3.5 hours. At the specified time point, 50 μL of PBS was added, mixed thoroughly, and then 400 μL of acetonitrile containing internal standard (IS, 100nM alprazolam, 500nM labetalol and 2μM ketoprofen) was added to precipitate the protein. The sample was vortexed for 5 minutes and then centrifuged at 20,000g for 15 minutes. The 0.5 hour sample was a non-centrifuged control sample.

8. After ultracentrifugation, take 50 μL from the center of the ultracentrifuge tube as the post-ultracentrifugation sample, and then add 50 μL of blank plasma to the sample. Then treat the sample the same as the non-centrifuged control sample. Take 100 μL of the supernatant to the injection plate, add 100 μL of ultrapure water and mix well for LC-MS/MS analysis.

9. Data Analysis: All calculations were performed using Microsoft Excel. The percentage of binding of the test compound and the control compound was calculated as follows:

Free rate % = (peak area ratio of _{ultracentrifuged samples} /peak area ratio of _{non-centrifuged control samples} ) × 100%

Binding rate % = 100% - free rate %

Table 5 Protein binding rate of the disclosed compounds in CD-1 mouse/human plasma

Test Example 5 Determination of membrane permeability and transport properties of the disclosed compounds

The membrane permeability and transport properties of the disclosed compounds were determined using the following test methods.

1. Test materials and instruments

1. Caco-2 cells (ATCC)

2. HEPES (Solarbio 804D049), Penicillin/Streptomycin (Solarbio 20200109) and Trypsin/EDTA (Solarbio), PBS (Solarbio 20200620)

3. Fetal bovine serum (FBS) (Sigma WXBD0055V), fluorescent yellow (Sigma MKCJ3738), NaHCO ₃ (Sigma SLBZ4647)

4. Hank’s Balanced Salt Solution (HBSS) (Gibco 2085528) and Non-Essential Amino Acids (NEAA) (Gibco 2211548), Trypsin/EDTA (Gibco 2120732)

5. High glucose DMEM medium (Corning 20319014)

6.96-well HTS transwell plate(Corning,3391)

7. Resistance tester (Millipore, ERS-2)

8. Vision(Nexcelom Bioscience)

9. Infinite 200PRO ELISA reader (Tecan, Infinite M200PRO)

10. Positive control compounds: metoprolol (Sinopharm 100084-201403), erythromycin (MCE 84550), and cimetidine (Sinopharm 100158-201406)

11. Kanamycin (Merck QR11795)

12. Triple QuadTM 5500+ LC-MS (AB Inc)

13.CO ₂ incubator (Thermo 371)

14.T-75 culture flask (Thermo 156499)

2. Test steps

1. Caco-2 cell culture

1) Preparation of transport buffer (HBSS containing 25 mM HEPES, pH 7.4): Accurately weigh 5.958 g HEPES and 0.35 g NaHCO ₃ , add 900 mL pure water to dissolve them, then add 100 mL 10× HBSS and stir evenly, adjust the pH to 7.4 with sodium hydroxide, and pass through filter.

2) Preparation of Caco-2 cell culture medium: FBS, penicillin, streptomycin, kanamycin and NEAA were added to high-glucose DMEM culture medium to prepare a cell culture medium containing 10% FBS, 0.1 mg/mL streptomycin, 100 units of penicillin, 0.6 μg/mL kanamycin and 1×NEAA.

3) Cultivate cells in a T-75 culture flask in a 37°C, 5% CO ₂ incubator. Discard the culture medium when the cells reach 80-90% density. Rinse the cells with 5 mL PBS, add 1.5 mL trypsin/EDTA, and then incubate in a 37°C incubator for 5-10 minutes until the cells fall off in a quicksand-like state. Finally, neutralize the trypsin/EDTA with Caco-2 cell culture medium.

4) Centrifuge the cell suspension at 120 x g for 10 minutes and discard the supernatant.

5) Add cell culture medium to resuspend the cells and adjust the density of the cell suspension to 6.86×10 ⁵ cells/mL.

2. Caco-2 Cell Seeding

1) Add 50 μL of culture medium to each well of the Transwell chamber, add 25 mL of Caco-2 cell culture medium to the lower layer, and place in a 37°C, 5% _CO2 incubator to preheat for 1 hour.

2) Add 50 μL of cell suspension to each well of the preheated Transwell chamber, and the final seeding density is 2.4×10 ⁵ cells/cm ² .

3) Culture for 14-18 days, changing the culture medium every other day, and changing the culture medium within 48 hours after the initial seeding. The culture medium must be changed the day before the experiment.

3. Assessing Monolayer Cell Membrane Integrity:

1) After 14 days of culture, the cells were confluent and differentiated and ready for transport experiments.

2) Use a resistor meter to measure the resistance of the single-layer membrane and record the resistance of each hole.

3) After the measurement, re-incubate the Transwell culture plate

4) Calculate TEER value:

TEER value = TEER (Ω) measured value × membrane area (cm ² )

The resistance of the cell monolayer membrane is <230Ω·cm ² , indicating that the cell monolayer membrane has poor compactness and cannot be used for the test.

4. Transport Experiment

1) Dilute the 10 mM DMSO stock solution of the disclosed compound or the positive control compound with DMSO to obtain a 2 mM stock solution, and then dilute the 2 mM stock solution with transport buffer to obtain a 10 μM working solution of the disclosed compound or the positive control compound.

2) Take out the Caco-2 cell plate from the incubator, wash the Transwell plate twice with preheated transport buffer, and then incubate it in a 37°C incubator for 30 minutes.

3) To determine the transport rate of the compound from the top to the base (A→B), add 108 μL of the working solution of the compound to the Transwell chamber (top), and immediately take out 8 μL of sample from the top to 72 μL of transport buffer, and add 240 μL of stop solution containing internal standard to stop the transport as the initial top sample. At the same time, add 300 μL of transport buffer to the receiving end (base). The experiment is set up with two samples.

4) To determine the transport rate of the compound from the base end to the top end (B→A), add 308 μL of the compound working solution to the receiving end (base end), and immediately take out 8 μL of sample from the base end to 72 μL of transport buffer, and add 240 μL of stop solution containing internal standard to stop the transport as the initial base end sample. At the same time, add 300 μL of transport buffer to the Transwell chamber (top end). The experiment is set up with two samples.

5) Place the cell culture plate in a 37°C CO ₂ incubator and incubate for 2 hours.

6) After the transport experiment, take 8 μL of sample from the dosing end (i.e., the top end in the A→B direction and the base end in the B→A direction) and add it to 72 μL of transport buffer, then add 240 μL of stop solution containing internal standard to terminate the transport. Take 80 μL of sample from the receiving end (i.e., the base end in the A→B direction and the top end in the B→A direction) and add it to 240 μL of stop solution containing internal standard, vortex at 1000 rpm for 10 minutes, and centrifuge at 3,220 g for 30 minutes. Take 100 μL of supernatant to the sample injection plate, add 100 μL of ultrapure water to mix, and use it for LC-MS/MS analysis.

7) After the transport experiment, measure the fluorescence value, prepare 10mM fluorescein stock solution with water, and then dilute it to 100μM with transport buffer. Add 100μL of fluorescein solution to the Transwell chamber (top), add 300μL of transport buffer to the base, and incubate in a _CO2 incubator at 37°C for 30 minutes. Take 80μL of solution from the base to a 96-well plate, and measure the cell fluorescence value (to detect membrane integrity) with an ELISA reader at an excitation wavelength of 485nm and an emission wavelength of 530nm.

The fluorescence value of Caco-2 cell monolayer was calculated using the following formula:

LY Leakage＝{I _acceptor ×0.3/(I _acceptor ×0.3+I _donor ×0.1)}×100%

I _acceptor refers to the fluorescence density on the receiving side (0.3 mL), and I _donor refers to the fluorescence density on the donating side (0.1 mL). LY>1.0% indicates poor cell monolayer membrane tightness, and the corresponding results will be excluded from the evaluation.

The peak areas of the compound on the dosing side and the receiving side were measured. The apparent permeability coefficient (P _app , unit: cm/s) and efflux ratio (ER) of the compound were calculated:

P _app ={V _A ×[drug] _acceptor /(Area×incubation time×[drug] _{initial dono} }

_VA is the volume of the receiving end solution (A→B is 0.3 mL, B→A is 0.1 mL), Area is the Transwell-96 plate membrane area (0.143 cm ² ); incubation time is the incubation time (unit: s).

P _app(BA) is the apparent permeability from the base to the apex; P _app(AB) is the apparent permeability from the apex to the base.

The calculated apparent permeability coefficient and efflux ratio of the disclosed compound are shown in Table 6. The test data show that the disclosed compound has good Caco-2 permeability.

Table 6 Apparent permeability coefficient and efflux ratio of the compounds disclosed herein

Test Example 6 Inhibitory Effects of the Disclosed Compounds on CYP2C9, CYP2D6, and CYP3A4 Enzyme Activities

The inhibition of the compounds on the enzyme activities of CYP2C9, CYP2D6, and CYP3A4 was determined using the following test method.

1. Test materials and instruments

1. Human liver microsomes (Corning 452117)

2.Na ₂ HPO ₄ (Shanghai Test 20040618)

3. KH ₂ PO ₄ (Shanghai Test No. 10017608)

4.NADPH (Roche diagnostics gmbh 10107824001)

5. Positive substrates: diclofenac sodium (Dalian Meilun MB1277), dextromethorphan hydrobromide (Dalian Meilun MB1568) and midazolam (Cerilliant M-908)

6. Positive inhibitors sulfaphenazole (Dalian Meilun MB3276), quinidine (Dalian Meilun MB1702) and ketoconazole (Dalian Meilun MB1132)

7. Tolbutamide (Sigma-aldrich T0891-100G)

8. AB Sciex Triple Quad 5500+ LC/MS

Two test steps

1. Preparation of 100 mM phosphate buffer (PBS): Weigh 7.098 g Na ₂ HPO ₄ , add 500 mL pure water and ultrasonically dissolve, as solution A. Weigh 3.400 g KH ₂ PO ₄ , add 250 mL pure water and ultrasonically dissolve, as solution B. Place solution A on a stirrer and slowly add solution B until the pH reaches 7.4 to prepare 100 mM PBS buffer.

2. Prepare 10 mM NADPH solution with 100 mM PBS buffer. Dilute 10 mM stock solution of the disclosed compound with DMSO to obtain 200× concentration of compound working solution (10000, 3333.3, 1111.1, 370.37, 123.46, 41.15, 13.7, 0 μM). Dilute positive inhibitor stock solution with DMSO to obtain 200× concentration of positive inhibitor working solution (sulfaphenazole, 1000, 300, 100, 30, 10, 3, 0 μM; quinidine/ketoconazole, 100, 30, 10, 3, 1, 0.3, 0 μM). Prepare 200× concentration of substrate working solution (1200 μM diclofenac, 400 μM dextromethorphan and 200 μM midazolam) with water, acetonitrile or acetonitrile/methanol.

3. Take 2μL 20mg/ml liver microsome solution, 1μL substrate working solution, 1μL compound working solution and 176μL PBS buffer, mix well, and pre-incubate in a 37℃ water bath for 10 minutes. Add 1μL positive inhibitor working solution to the positive control group instead of the compound working solution. At the same time, pre-incubate 10mM NADPH solution in a 37℃ water bath for 10 minutes. After 10 minutes, take 20μL NADPH and add it to each well to start the reaction. Incubate at 37℃ for 5 minutes (CYP2C9), 20 minutes (CYP2D6) or 5 minutes (CYP3A4). All incubation samples are set up as double samples. After incubation for the corresponding time, add 400μL ice methanol containing internal standard (50ng/mL tolbutamide) to all samples to terminate the reaction. Vortex to mix, centrifuge at 3220g and 4℃ for 30 minutes. After centrifugation, transfer 50 μL of supernatant to the sample injection plate, add 100 μL of ultrapure water and mix well for LC-MS/MS analysis.

The IC ₅₀ values of the disclosed compounds against CYP2C9, CYP2D6 and CYP3A4 were calculated by Excel XLfit 5.3.1.3 and are shown in Table 7. The test data show that the disclosed compounds have no significant inhibitory effect on CYP2C9, CYP2D6 and CYP3A4.

Table 7 _IC50 values of the disclosed compounds against CYP2C9, CYP2D6 and CYP3A4

Test Example 7 Detection of the pharmacokinetic properties of the disclosed compounds in mice

1. Test material

1. Healthy adult CD-1 male mice were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

2. PEG400 (polyethylene glycol 400), HP-β-CD (hydroxypropyl-β-cyclodextrin), hydrochloric acid, tolbutamide, methanol, acetonitrile, propylene glycol and formic acid were purchased from Merck (USA).

3. K ₂ EDTA anticoagulant tube was purchased from Jiangsu Xinkang Medical Instrument Co., Ltd.

2. Test Methods

1. Drug preparation: Weigh a certain amount of the disclosed compound and dissolve it in 20% PEG400 + 4% 1M HCl + 76% (20% HP-β-CD) to prepare a 1 mg/mL solution for oral administration. Dilute the oral administration solution with 20% HP-β-CD to 0.2 mg/mL for intravenous administration.

2. Administration method: Oral gavage group: CD-1 mice were given oral gavage without fasting, and the dosage was 10 mg/kg. Intravenous group: CD-1 mice were given intravenous administration without fasting, and the dosage was 1 mg/kg.

3. Animal experiment: After oral or intravenous administration to mice, more than 20 μL of blood was collected from the eye socket at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h after administration and added to K ₂ EDTA anticoagulant tubes. The plasma was separated by centrifugation at 12000 rpm, 4°C for 5 minutes and stored at -20°C.

4. Determine the content of the test compound in mouse plasma after oral administration of different concentrations of drugs: melt the sample at room temperature, vortex for 1 min; quantitatively transfer 10 μL to a 2 mL 96-well plate, add 50 μL of internal standard solution (10 ng/mL tolbutamide acetonitrile solution) and 50 μL of precipitant (acetonitrile: methanol 7:3), oscillate (1000 rpm×3 min); centrifuge (4000 rpm×15 min), transfer 80 μL of supernatant to a 2 mL 96-well plate, add 80 μL of diluent (water), and oscillate (1000 rpm×3 min). Quantitative detection was performed using an LC-MS/MS system (AB Sciex Triple Quad 6500+).

5. Data processing: The pharmacokinetic parameters were calculated using the non-compartmental statistical moment method of Phoenix WinNonlin 8.0 software (Certara, USA). The test results are shown in Tables 8 and 9. The test data show that the disclosed compounds have good oral absorption in mice.

Table 8 Pharmacokinetic parameters of the compounds after single oral administration in mice

Note: NA stands for Not Applicable.

Table 9 Pharmacokinetic parameters of the compound after single intravenous administration in mice

Test Example 8 Detection of Pharmacokinetic Properties of the Disclosed Compounds in Rats

1. Test material

1. Healthy adult SD male rats were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

2.PEG400 (polyethylene glycol 400), HP-β-CD (hydroxypropyl-β-cyclodextrin), hydrochloric acid, tolbutamide, methanol, acetonitrile, propylene glycol Diols and formic acid were purchased from Merck (USA).

3. K ₂ EDTA anticoagulant tube was purchased from Jiangsu Xinkang Medical Instrument Co., Ltd.

2. Test Methods

1. Drug preparation: Weigh a certain amount of the disclosed compound and dissolve it in 20% PEG400 + 4% 1M HCl + 76% (20% HP-β-CD) to prepare a 1 mg/mL solution for oral administration. Dilute the oral administration solution with 20% HP-β-CD to 0.2 mg/mL for intravenous administration.

2. Administration method: Oral gavage group: SD rats were fasted and the dosage was 10 mg/kg. Intravenous group: SD rats were fasted and the dosage was 1 mg/kg.

3. Animal experiment: After oral or intravenous administration to rats, 100 μL of blood was collected from the jugular vein at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h after administration and added to an anticoagulant tube. The plasma was separated by centrifugation at 12000 rpm, 4°C for 5 minutes and stored at -20°C.

4. Determine the content of the test compound in rat plasma after oral or intravenous administration of different concentrations of drugs: take the sample and melt it at room temperature, vortex for 1 minute; quantitatively transfer 10μL to a 2mL 96-well plate, add 50μL internal standard solution (10ng/mL tolbutamide acetonitrile solution) and 50μL precipitant (acetonitrile: methanol 7:3), oscillate (1000rpm×3min); centrifuge (4000rpm×15min), transfer 80μL of supernatant to a 2mL 96-well plate, add 80μL diluent (water), and oscillate (1000rpm×3min). Quantitative detection was performed using an LC-MS/MS system (AB Sciex Triple Quad 6500+).

5. Data processing: The pharmacokinetic parameters were calculated using the non-compartmental statistical moment method of Phoenix WinNonlin 8.0 software (Certara, USA). The test results are shown in Tables 10 and 11. The test data show that the disclosed compounds have good oral absorption in rats.

Table 10 Pharmacokinetic parameters of the compounds after single oral administration in rats

Table 11 Pharmacokinetic parameters of the compounds after single intravenous administration in rats

Claims (29)

A compound represented by formula (I) or a pharmaceutically acceptable salt thereof,
TL-Linker-DIM
(I) in: The DIM is a ligand compound capable of binding to E3 ubiquitin ligase; The Linker is a linking group that covalently binds at least one TL and at least one DIM; The TL is a group as shown below:
in: A ¹ , A ² , A ³ , A ⁴ are independently selected from CH, CR ¹ or N; R ¹ is selected from deuterium, halogen, CN, -C(O)NR ^1a R ^1b , NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₀ aryl, 5-10 membered heteroaryl or 4-14 membered heterocyclyl, wherein the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₆ -C ₁₀ aryl, 5-10 membered heteroaryl or 4-14 membered heterocyclyl is optionally substituted with one or more ^Ra ; R ^1a is selected from H, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl, wherein the C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl is optionally substituted by one or more R ^1c ; R ^1b is selected from H or C ₁ -C ₆ alkyl; One of B ¹ and B ² is selected from C-NHR ² , C-OR ² or C-CHR ^2a R ^2b , and the other is selected from CH or N; R ² , R ^2a , R ^2b are independently selected from H, deuterium, halogen, CN, NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-8 membered heterocyclyl, phenyl or 5-6 membered heteroaryl, wherein the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-8 membered heterocyclyl, phenyl or 5-6 membered heteroaryl is optionally substituted by one or more R ^b ; Ring Q is a 5-10 membered heteroaromatic ring, wherein the 5-10 membered heteroaromatic ring is optionally substituted by one or more R ^c ; One of B ³ and B ⁴ is C and is connected to ring Q, and the other is selected from CH or N; X is selected from S, O or NR ³ ; R ³ is selected from H, deuterium, halogen, CN, NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, phenyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl, wherein the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, phenyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl is optionally substituted with one or more R ^d ; ^Ra , ^Ric , ^Rb , ^Rc , ^Rd are independently selected from deuterium, halogen, OH, CN, _NH2 , =O, _C1 - _C6 alkyl, _C3 - _C6 cycloalkyl, 5-6-membered heteroaryl or 4-8-membered heterocyclyl, wherein the OH, _NH2 , _C1 - _C6 alkyl, C3- _C6 cycloalkyl, 5-6-membered heteroaryl or _4-8 -membered heterocyclyl is optionally substituted by one or more ^Re ; R ^e is selected from deuterium, halogen, OH, NH ₂ , ═O, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, CN, COOH, C(O)(C ₁ -C ₃ alkyl), CONH ₂ , C(O)O(C ₁ -C ₃ alkyl), S(O) ₂ (C ₁ -C ₃ alkyl), C ₃ -C ₆ cycloalkyl or 4-6 membered heterocyclyl. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein one of ^B1 and ^B2 is C- ^NHR2 and the other is selected from CH or N; or, ^B1 is C- ^NHR2 and ^B2 is selected from CH or N; or, ^B1 is C- ^NHR2 and ^B2 is N. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein ^B3 is C and is connected to ring Q, and ^B4 is selected from CH or N; or, ^B3 is C and is connected to ring Q, and ^B4 is CH; or, ^B3 is selected from CH or N, and ^B4 is C and is connected to ring Q. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein R ² , R ^2a , and R ^2b are independently selected from H, deuterium, OH, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-7 membered heterocyclyl, phenyl or 5-6 membered heteroaryl, and the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-7 membered heterocyclyl, phenyl or 5-6 membered heteroaryl is optionally substituted by one or more R ^b . The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein B ¹ is C-NHR ² , R ² is selected from H, deuterium, OH, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-7 membered heterocyclyl, phenyl or 5-6 membered heteroaryl, and the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, 4-7 membered heterocyclyl, phenyl or 5-6 membered heteroaryl is optionally substituted by one or more R ^b ; or, B ¹ is C-NHR ² , R ² is selected from H, deuterium, OH, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl or 4-7 membered heterocyclyl, and the OH, NH ₂ , C ₁ -C 6 alkyl, C 3 -C 6 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by one or more R ^b ; or, B ¹ is C-NHR ² , R 2 is selected from H, deuterium, OH, C 1 -C ₆ alkyl, C _{3 -C 6} cycloalkyl or 4-7 membered heterocyclyl ² is selected from H, deuterium, OH or C ₁ -C ₆ alkyl, The OH, _NH2 or _C1 - _C6 alkyl is optionally substituted by one or more ^Rb ; or, ^B3 is C and connected to ring Q, ^B4 is selected from CH or N; or, ^B1 is C- ^NHR2 , ^R2 is selected from tetrahydropyranyl, oxetanyl or _C1 - _C4 alkyl optionally substituted by ^Rb . The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein ^A1 and ^A4 are selected from CH, ^CR1 or N, and ^A2 and ^A3 are selected from CH or ^CR1 ; or, ^A4 is selected from CH, ^CR1 or N, and ^A1 , ^A2 and ^A3 are selected from CH or ^CR1 ; or, ^A4 is selected from CH or N, ^A2 is ^CR1 , and ^A1 and ^A3 are both CH; or, ^A4 is selected from CH, ^A2 is ^CR1 , and ^A1 and ^A3 are both CH; or, ^A1 is selected from CH, ^CR1 or N, and ^A2 , ^A3 and ^A4 are selected from CH or ^CR1 ; or, ^A1 is selected from CH or N, ^A3 is ^CR1 , and ^A2 and ^A4 are both CH; or, ^A1 is selected from CH, ^A3 is ^CR1 , and ^A2 and ^A4 are both CH. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein R ¹ is selected from deuterium, halogen, CN, -C(O)NR ^1a R ^1b , NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl or 4-7 membered heterocyclyl, and the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by one or more ^Ra ; or, R ¹ is selected from deuterium, halogen, CN, NO ₂ , OH, NH ₂ , C 1 -C ₆ alkyl, C ₃ -C ₆ cycloalkyl or 4-7 membered heterocyclyl, and the OH, NH 2 , C ₁ -C _{6 alkyl, C 3 -C 6 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by one or more Ra; or, R 1 is selected from deuterium, halogen, CN, NO 2 , OH, NH 2 , C 1 -C 6} ^{alkyl ,} _{C 3 -C 6 cycloalkyl} or 4-7 membered heterocyclyl is optionally substituted by one or more Ra; -C ₄ alkyl, said OH or C ₁ -C ₄ alkyl is optionally substituted by one or more ^Ra . The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein R ^1a is selected from H or C ₁ -C ₆ alkyl, and the C ₁ -C ₆ alkyl is optionally substituted by one or more R ^1c ; R ^1b is selected from H or C ₁ -C ₆ alkyl; or, R ^1a is selected from C ₃ -C ₆ cycloalkyl, 5-6-membered heteroaryl or 4-7-membered heterocyclyl, and the C ₃ -C ₆ cycloalkyl, 5-6-membered heteroaryl or 4-7-membered heterocyclyl is optionally substituted by one or more R ^1c ; R ^1b is selected from H or C ₁ -C ₆ alkyl. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, wherein ring Q is a 5-6 membered heteroaromatic ring optionally substituted by one or more R ^c ; or, ring Q is a 5 membered heteroaromatic ring optionally substituted by one or more R ^c , such as the following groups optionally substituted by one or more R ^c : pyrazolyl, imidazolyl, triazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl or thiadiazolyl; or, ring Q is the following groups optionally substituted by one or more R ^c : thiadiazolyl, triazolyl, pyrazolyl. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, wherein R ³ is selected from H, deuterium, halogen, CN, NO ₂ , OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, and the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl are optionally substituted by one or more R ^d ; or, R ³ is selected from H, deuterium, C ₁ -C ₆ alkyl or C ₃ -C ₆ cycloalkyl, and the C ₁ -C ₆ alkyl or C ₃ -C ₆ cycloalkyl are optionally substituted by one or more R ^d ; or, R ³ is selected from H, deuterium or C ₁ -C ₄ alkyl optionally substituted by one or more R ^d . The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 10, wherein X is selected from S, O or NH; or X is selected from NR ³ ; or X is selected from NH. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein ^Ra , ^R1c , ^Rb , ^Rc , and ^Rd are independently selected from deuterium, halogen, OH, CN, _NH2 , =O, _C1 - _C6 alkyl, _C3 - _C6 cycloalkyl, or 4-7-membered heterocyclyl, and the OH, _NH2 , _C1 - _C6 alkyl, _C3 - _C6 cycloalkyl, or 4-7-membered heterocyclyl is optionally substituted by one or more ^Re ; or, ^Ra , ^R1c , ^Rb , ^Rc , and ^Rd are independently selected from deuterium, halogen, OH, CN, _NH2 , =O, or _C1 - _C6 alkyl, and the OH, _NH2 , or _C1 - _C6 alkyl is optionally substituted by one or more ^Re . The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein ^Re is selected from deuterium, halogen, OH, _NH2 , _C1 - _C3 alkyl, _C1 - _C3 alkoxy, COOH, C(O)( _C1 - _C3 alkyl), _CONH2 , C(O)O( _C1 - _C3 alkyl), _C3 - _C6 cycloalkyl or 4-6 membered heterocyclyl; or, ^Re is selected from halogen, _C1 - _C3 alkyl, _C1 - _C3 alkoxy, COOH, C(O)( _C1 - _C3 alkyl), _CONH2 or C(O)O( _C1 - _C3 alkyl). The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 13, wherein the TL is selected from the structure shown in the following (TL-1):
wherein B ² and B ⁴ are independently selected from CH or N, and A ¹ , A ² , A ³ , A ⁴ , X, R ² and Q are as defined in any one of claims 1-13. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 13, wherein the TL is selected from the structure shown in the following (TL-2):
wherein: B ² and B ⁴ are independently selected from CH or N, A ⁴ is selected from CH or N, n is selected from 0, 1 or 2, and R ¹ , R ² , X and ring Q are as defined in any one of claims 1-13. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 15, wherein the TL is selected from one of the following structures: The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 16, wherein the Linker is a linking group that covalently binds a TL and a DIM. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 17, wherein the linker is selected from: -L ^A -, -L ^B -, -R ^1L -, -R ^2L -, -Q ¹ -, -Q ² -, or The Linker is selected from Linker or The Linker is wherein: ^-LA- , ^-LB- are independently selected from a chemical bond, -O-, -S-, -NR 3'-, -CR ^4'R5'- , -CR 4'R5' ^- NR ^3'- , -CR ^{4'R5' -} O-, ^{-C(O)-, -CR 4'R5' - C} (O)-, -S(O)-, -S(O) ^2- , -C(S ⁾ -, -C(O)O- or -C(O)NR ^6'- _; R ^1L and R ^2L are independently selected from a chemical bond, -C(O)-, alkylene, heteroalkylene, alkenylene and alkynylene, wherein the alkylene, heteroalkylene, alkenylene and alkynylene are optionally substituted by a group selected from the group consisting of halogen, alkyl, alkoxy, haloalkyl, OH, hydroxyalkyl, CN, NH ₂ , =O, cycloalkyl, heterocyclyl, aryl, heteroaryl; Q ¹ , Q ² , Q ³ and Q ⁴ are independently selected from cycloalkylene, heterocyclylene, arylene, heteroarylene or cycloalkenylene, wherein the cycloalkylene, heterocyclylene, arylene, heteroarylene and cycloalkenylene are each independently optionally substituted by a group selected from the following: halogen, alkyl, alkoxy, haloalkyl, OH, hydroxyalkyl, CN, NH ₂ , =O, cycloalkyl, heterocyclyl, aryl, heteroaryl; R ^3′ is selected from H, alkyl, heteroalkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R ^4′ and R ^5′ are each independently selected from H, halogen, alkyl, alkoxy, haloalkyl, OH, hydroxyalkyl, CN, NH ₂ , ═O, cycloalkyl, heterocyclyl, aryl or heteroaryl; R ^6′ is selected from H, alkyl, heteroalkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18, wherein the DIM is selected from a ligand compound capable of binding to a cereblon-type E3 ubiquitin ligase. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 19, wherein the DIM is selected from the structure shown in formula (DIM-1) or (DIM-2):
in: Selected from Y is a chemical bond, or Y is selected from _YA , O, NH, NR _E , C(O)O, C(O)NR _E ', NR _E'C (O), YA- _NH , _YA -NR _E , _{YA- C} (O), YA- _C (O)O, YA- _OC (O), YA-C(O)NR _E ' or YA- _{NR E'C} (O), wherein _YA is selected from _C1 - _C6 alkylene, _C2 - _C6 alkenylene or _C2 - _C6 alkynylene; X' is selected from C(O) or C( _RA ) ₂ ; XA _{- XB} is selected from C( _RA )=N or C( _RA ) ₂ -C( _RA ) ₂ ; Each _RA is independently selected from H or C ₁ -C ₃ alkyl, wherein the C ₁ -C ₃ alkyl is optionally substituted with C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl; Each _RA ' is independently selected from _C1 - _C3 alkyl; Each _RB is independently selected from H or _C1 - _C3 alkyl, or two _RBs together with the atoms to which they are attached form C(O), _C3 - _C6 cycloalkyl, _C3 - _C6 cycloalkenyl or 4-6 membered heterocyclyl; R _C is selected from H, halogen or C ₁ -C ₃ alkyl; Each R _D is independently selected from halogen, NO ₂ , NH ₂ , OH, COOH, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy; Each _RE is independently selected from C1- _C6 alkyl, _C2 - _C6 alkenyl, _C3 - _C8 cycloalkyl, 3-8 membered heterocycloalkyl, C(O)-C1- _C6 alkyl _, C(O) _-C2 - _C6 alkenyl, C(O) _{-C3 -C8 cycloalkyl} or C(O)-3-8 membered heterocycloalkyl, and the _RE is optionally substituted by a group selected from halogen, N(R _a ) ₂ , NHC(O)R _a , NHC(O)OR _a , OR _b , C ₃ -C ₈ cycloalkyl, 3-8 membered heterocycloalkyl, C ₆ -C ₁₀ aryl, or 5-10 membered heteroaryl, wherein the C ₃ -C ₈ cycloalkyl, 3-8 membered heterocycloalkyl, C ₆ -C ₁₀ aryl, or 5-10 membered heteroaryl is optionally further substituted by a group selected from the group consisting of halogen, NH ₂ , CN, NO ₂ , OH, COOH, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, or C ₁ -C ₆ haloalkoxy; R _E 'is selected from H, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₃ -C ₈ cycloalkyl or 3-8 membered heterocycloalkyl, wherein the C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₃ -C ₈ cycloalkyl or 3-8 membered heterocycloalkyl is optionally substituted by a group selected from the group consisting of halogen, N(R _a ) ₂ , NHC(O)R _a , NHC(O)OR _a , OR _b , C ₃ -C ₈ cycloalkyl, 3-8 membered heterocycloalkyl, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl, wherein the C ₃ -C _{8 cycloalkyl, 3-8 membered heterocycloalkyl, C 6 -C 10 aryl or 5-10 membered heteroaryl is optionally further substituted by a group selected from the group consisting of halogen, NH 2 , CN, NO 2 , OH, COOH, C 1 -C 6 alkyl, C 1 -C 6 alkenyl, C 3 -C 8 cycloalkyl or 3-8 membered} heterocycloalkyl C ₁ -C ₆ haloalkyl, C 1 -C ₆ alkoxy or C ₁ -C ₆ haloalkoxy; Each _Ra is independently selected from H or _C1 - _C6 alkyl; R _b is selected from H or p-toluenesulfonyl; t is selected from 0 or 1; m is selected from 0, 1, 2 or 3; p is selected from 0, 1 or 2. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 19, wherein the DIM is selected from the structure represented by formula (DIM-11):
in: _XC is selected from a chemical bond, _-CH2- , -CHCF3-, _-SO2- , _-S (O)-, -P(O)R'-, -P(O)(OR')-, -P(O)( _NR'2 )-, -C(O)-, -C(S)- or _XD is selected from C, N or Si; X _E is selected from a chemical bond, -CR' ₂ -, -NR'-, -O-, -S- or -SiR' ₂ -; _RF is absent or is selected from H, _deuterium , halogen, CN, -OR'-, -SR'-, -S(O)R'-, -S(O)2R'-, -NR'2-, -P(O)(OR') ₂ , -P(O)( _{NR'2 )} OR'-, -P(O)( _NR'2 ) ₂ , -Si(OH) _2R ', -Si(OH) _R'2 , _-SiR'3 , or _C1 - _C4 alkyl _; Each _RG is independently selected from H, deuterium, _RH , halogen, CN, -NO2, _-OR ', -SR', _-NR'2 , _-SiR'3 , -S(O) _2R ', -S(O) _2NR'2 , -S(O)R _' , -C(O)R', -C(O)OR', -C(O) _NR'2 , -C(O)N(R')OR', -C(R') _2N (R')C(O)R', -C(R') _2N (R')C(O) _NR'2 , -OC(O)R', -OC(O) _NR'2 , -OP(O)R'2, -OP(O)(OR') ₂ , _-OP (O ₎ (OR')NR'2, -OP(O)( _NR'2 ) ₂ , -N(R')C(O)OR', -N(R')C(O)R', -N(R')C(O)NR' ₂ , -N(R')S(O) ₂ R' , -N(R')P(O)R' ₂ , -N(R')P(O)(OR') ₂ , -N(R')P(O)(OR')NR' ₂ or -N(R')P(O)(NR' ₂ ) ₂ ; Each _RH is independently selected from _C1 - _C6 alkyl, phenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl; Ring E, Ring F, Ring G are independently selected from phenyl, 6-membered heteroaryl, C ₅ -C ₇ cycloalkyl, C ₅ -C ₇ cycloalkenyl, 5-7-membered heterocyclyl or 5-6-membered heteroaryl, wherein Ring E, Ring F and Ring G are each optionally further substituted by =O; L ¹ is selected from a chemical bond, a C ₁ -C ₃ alkylene group, a C ₂ -C ₃ alkenylene group or a C ₂ -C ₃ alkynylene group, wherein any _{one or} two methylene groups in the C ₁ -C ₃ alkylene group, the C ₂ -C ₃ alkenylene group or the C 2 -C 3 alkynylene group are optionally replaced by the following groups: -O-, -C(O)-, -C(S)-, -C(R') ₂ -, -CH(R')-, -C(F) ₂ -, -N(R')-, -S- or -S(O) ₂ -; Each R' is independently selected from H, C ₁ -C ₆ alkyl, phenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl, or two R' and the atoms to which they are attached together form a 4-7 membered heterocyclyl or 5-6 membered heteroaryl; q is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 19, wherein the DIM is selected from the structure shown in formula (DIM-12):
Wherein: Ring H is selected from C ₅ -C ₉ cycloalkyl, C ₅ -C ₉ cycloalkenyl or 5-9 membered heterocyclyl, wherein said C ₅ -C ₉ cycloalkyl, C ₅ -C ₉ cycloalkenyl or 5-9 membered heterocyclyl is optionally substituted by =O; k is selected from 0, 1, 2, 3 or 4; _XC is selected from a chemical bond, _-CH2- , -CH( _CF3 )-, _-SO2- , -S(O)-, -P(O)R'-, -P(O)(OR')-, -P(O)( _NR'2 )-, -C(O)-, -C(S)- or _XD is selected from C, N or Si; X _E is selected from a chemical bond, -CR' ₂ -, -NR'-, -O-, -S- or -SiR' ₂ -; _RF is absent or is selected from H, _deuterium , halogen, CN, -OR', -SR', -S(O)R', -S(O) _2R ', -NR'2, -P(O)(OR') ₂ , -P(O)( _NR'2 )OR', -P(O)( _NR'2 ) ₂ , -Si(OH) _2R ', -Si(OH) _R'2 , _-SiR'3 , or _C1 - _C4 alkyl _; Each _RG is independently selected from H, deuterium, _RH , halogen, CN, -NO2, _-OR ', -SR', _-NR'2 , _-SiR'3 , -S(O) _2R ', -S(O) _2NR'2 , -S(O)R _' , -C(O)R', -C(O)OR', -C(O) _NR'2 , -C(O)N(R')OR', -C(R') _2N (R')C(O)R', -C(R') _2N (R')C(O) _NR'2 , -OC(O)R', -OC(O) _NR'2 , -OP(O)R'2, -OP(O)(OR') ₂ , _-OP (O ₎ (OR')NR'2, -OP(O)( _NR'2 ) ₂ , -N(R')C(O)OR', -N(R')C(O)R', -N(R')C(O)NR' ₂ , -N(R')S(O) ₂ R' , -N(R')P(O)R' ₂ , -N(R')P(O)(OR') ₂ , -N(R')P(O)(OR')NR' ₂ or -N(R')P(O)(NR' ₂ ) ₂ ; L ¹ is selected from a chemical bond, a C ₁ -C ₃ alkylene group, a C ₂ -C ₃ alkenylene group or a C ₂ -C ₃ alkynylene group, wherein any _{one or} two methylene groups in the C ₁ -C ₃ alkylene group, the C ₂ -C ₃ alkenylene group or the C 2 -C 3 alkynylene group are optionally replaced by the following groups: -O-, -C(O)-, -C(S)-, -C(R') ₂ -, -CH(R')-, -C(F) ₂ -, -N(R')-, -S- or -S(O) ₂ -; Ring E is selected from phenyl, 5-6 membered heteroaryl, C ₅ -C ₇ cycloalkyl, C ₅ -C ₇ cycloalkenyl or 5-7 membered heterocyclyl, and the ring E is optionally further substituted by =O; Each R' is independently selected from H, _C1 - _C6 alkyl, phenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl, or two R' and the atoms to which they are attached together form a 4-7 membered heterocyclyl or 5-6 membered heteroaryl. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 19, wherein the DIM is selected from the structure represented by formula (DIM-13):
wherein k is selected from 0, 1, 2, 3 or 4; _XC is selected from a chemical bond, _-CH2- , -CH( _CF3 )-, _-SO2- , -S(O)-, -P(O)R'-, -P(O)(OR')-, -P(O)( _NR'2 )-, -C(O)-, -C(S)- or _XD is selected from C, N or Si; X _E is selected from a chemical bond, -CR' ₂ -, -NR'-, -O-, -S- or -SiR' ₂ -; _RF is absent or is selected from H, _deuterium , halogen, CN, -OR', -SR', -S(O)R', -S(O) _2R ', -NR'2, -P(O)(OR') ₂ , -P(O)( _NR'2 )OR', -P(O)( _NR'2 ) ₂ , -Si(OH) _2R ', -Si(OH) _R'2 , _-SiR'3 , or _C1 - _C4 alkyl _; Each _RG is independently selected from H, deuterium, _RH , halogen, CN, -NO2, _-OR ', -SR', _-NR'2 , _-SiR'3 , -S(O) _2R ', -S(O _{)2NR'2 ,} -S(O)R', -C(O)R', -C(O)OR', -C(O)NR' ₂ , -C(O)N(R')OR' , -C(R') ₂ N(R')C(O)R' , -C(R') ₂ N(R')C(O)NR' ₂ , -OC(O)R' , -OC(O)NR' ₂ , -OP(O)R' ₂ , -OP(O)(OR') ₂ , -OP(O)(OR')NR' ₂ , -OP(O)(NR' ₂ ) ₂ , -N(R')C(O)OR', -N(R')C(O)R', -N(R')C(O)NR' ₂ , -N(R')S(O) ₂ R', -N(R')P(O)R' ₂ , -N(R')P(O)(OR') ₂ , -N(R')P(O)(OR')NR' ₂ or -N(R')P(O)(NR' ₂ ) ₂ ; L ¹ is selected from a chemical bond, a C ₁ -C ₃ alkylene group, a C ₂ -C ₃ alkenylene group or a C ₂ -C ₃ alkynylene group, wherein any _{one or} two methylene groups in the C ₁ -C ₃ alkylene group, the C ₂ -C ₃ alkenylene group or the C 2 -C 3 alkynylene group are optionally replaced by the following groups: -O-, -C(O)-, -C(S)-, -C(R') ₂ -, -CH(R')-, -C(F) ₂ -, -N(R')-, -S- or -S(O) ₂ -; Ring E is selected from phenyl, 5-6 membered heteroaryl, C ₅ -C ₇ cycloalkyl, C ₅ -C ₇ cycloalkenyl or 5-7 membered heterocyclyl, and the ring E is optionally further substituted by =O; Each R' is independently selected from H, _C1 - _C6 alkyl, phenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl, or two R' and the atoms to which they are attached together form a 4-7 membered heterocyclyl or 5-6 membered heteroaryl. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein the compound has a structure selected from one of the following:

A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 24, and a pharmaceutically acceptable excipient. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 24 or a pharmaceutical composition according to claim 25 in the preparation of a medicament for preventing or treating a disease mediated by IRAK4. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 24, or a pharmaceutical composition according to claim 25 for preventing or treating a disease mediated by IRAK4. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 24, or a pharmaceutical composition according to claim 25, in preventing or treating a disease mediated by IRAK4. A method for treating a disease mediated by IRAK4 in a mammal, comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 24, or a pharmaceutical composition according to claim 25 to a mammal, preferably a human, in need of such treatment.