WO2023061478A1

WO2023061478A1 - Tricyclic compound

Info

Publication number: WO2023061478A1
Application number: PCT/CN2022/125361
Authority: WO
Inventors: 麦万笋; 刘晓武; 金京海; 祝伟; 邹昊; 李正涛
Original assignee: 先声再明医药有限公司
Priority date: 2021-10-15
Filing date: 2022-10-14
Publication date: 2023-04-20
Also published as: CN118201923A

Abstract

Provided in the present disclosure are a compound as represented by formula (I) and a pharmaceutically acceptable salt thereof, a composition thereof, and the use thereof in the preparation of a drug for treating diseases related to CRBN protein. The compound can further be used as a synthetic intermediate for a bifunctional compound for targeting protein degradation, and thus can be used for preparing a bifunctional compound for targeting protein degradation.

Description

Tricyclic compounds

This application claims priority to the following earlier application, which is hereby incorporated by reference in its entirety:

The patent application number submitted to the State Intellectual Property Office of China on October 15, 2021 is 202111206934.7, and the invention name is the prior application of "tricyclic compounds".

technical field

The disclosure belongs to the field of medical technology, and specifically relates to a tricyclic compound, its composition, and its application in the preparation of drugs for treating diseases related to CRBN protein. The compound is used to prepare bifunctional PROTAC compounds for treatment of related diseases.

Background technique

CRBN (Cereblon) is a protein encoded by the CRBN gene in the human body. CRBN is widely expressed in testis, spleen, prostate, liver, pancreas, placenta, kidney, lung, skeletal muscle, ovary, small intestine, peripheral blood leukocytes, colon, brain and retina, while in brain tissue (including retina) and testis The expression in was significantly higher than that in other tissues.

Studies have found that CRBN is closely related to the metabolism and proliferation of normal cells and tumor cells. On the one hand, its existence ensures the normal metabolic function and normal physiological function of the ion channel, which is very important for maintaining cell growth and proliferation. On the other hand, CRBN is involved in the occurrence of many diseases such as cancer. Shi et al., J. Immunol. Res. Article ID 9130608 (2017). As an important target of anti-tumor and immunomodulator drugs, CRBN has been proven to have clear curative effects in various hematological malignancies such as multiple myeloma and chronic lymphocytic leukemia, and autoimmune diseases such as systemic lupus erythematosus. However, the existing meridamine drugs have many side effects, especially peripheral neuropathy. At present, it is necessary to develop new CRBN modulator drugs to improve clinical treatment effect, reduce clinical side effects, and benefit long-term use of patients.

In addition, CRBN can interact with damaged DNA-binding protein 1 (DDB1), form an E3 ubiquitin ligase complex with Cullin 4 (Cul4A) and the E2-binding protein ROC1 (also known as RBX1), in which, CRBN acts as the substrate receptor of CRBN-CRL4, and after binding to the substrate, the substrate protein undergoes proteolysis through the ubiquitin-proteasome pathway. See, Chang et al., Int. J. Biochem. Mol. Biol. 2(3):287-94 (2011).

The ubiquitin-proteasome pathway (UPP) is a key pathway for regulating key regulatory proteins and degrading misfolded or abnormal proteins. Ubiquitin molecules tag proteins for proteasomal degradation through covalent attachment of E3 ubiquitin ligases to terminal lysine residues, where the protein is digested into small peptides and eventually into its constituent amino acids with as building blocks for new proteins. UPP is important for multiple cellular processes, and if defective or out of balance, it contributes to the pathogenesis of a variety of diseases. Defective proteasomal degradation has been shown to be associated with a variety of clinical conditions including Alzheimer's disease, Parkinson's disease, Huntington's disease, muscular dystrophy, cardiovascular disease and cancer, among others. PROTAC (Proteolysis targeting chimeras) is a new technology for targeting and degrading target proteins developed based on the cell's own ubiquitin-proteasome pathway (UPP). PROTAC molecules are bifunctional molecules that can not only bind to target proteins, but also recruit E3 ubiquitin ligase to ubiquitinate target proteins, and then degrade target proteins through proteasomes. Therefore, CRBN ligands can also be used to prepare bifunctional PROTAC (proteolysis-directed chimera) compounds for the treatment of related diseases. Therefore, it is necessary to further develop small molecular compounds that can regulate CRBN and degrade specific proteins for the treatment of tumors. In addition, it is still necessary to find CRBN ligands that can be used as therapeutic agents, and at the same time, CRBN ligands can be further used to prepare bifunctional PROTAC compounds for the treatment of diseases.

Contents of the invention

The disclosure provides a compound represented by formula (I) and a pharmaceutically acceptable salt thereof:

in,

Y ¹ , Y ² , Y ³ are independently selected from CR ² or N;

X ¹ is selected from C, CR ^1a or N;

X ² is selected from CR ^2a , CR ^2a R ^2b , N or NR ^2a ;

X ³ is selected from O, S, C(=O), C(=S), S(=O) ₂ , S(O), CR ^3a R ^3b or NR ^3a ;

X ⁴ is selected from O, S, C(=O), C(=S), S(=O) ₂ , S(O), CR ^4a R ^4b or NR ^4a ;

R ^1a , R ^2a , R ^2b , R ^3a , R ^3b , R ^4a , R ^4b are independently selected from H, halogen, CN, OH, NH ₂ , -C(O) ^Rf , -C(O)N( R ^f ) ₂ , -C(O)OR ^f , -C(O)ON(R ^f ) ₂ , -NHC(O)R ^f , -NHC(O)OR ^f , -NHC(O)N(R ^f ) ₂ , -S(O) ₂ (R ^f ), C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, C ₆ -C ₁₀ aryl, 4-10 membered heterocyclyl or 5-10 OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, C ₆ -C ₁₀ aryl, 4-10 membered heterocyclyl or 5-10 membered heteroaryl Aryl is optionally ^substituted by R;

Or R ^1a , R ^4a and the cyclic group they are connected to form a carbon bridged ring or a bridged heterocyclic ring;

Or R ^2a , R ^3a and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

Or R ^3a , R ^4a and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

Or R ^2a , R ^2b and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

Or R ^3a , R ^3b and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

Or R ^4a , R ^4b and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

R ¹ is independently selected from halogen, CN, OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, 4-8 membered heterocyclyl, C ₆ -C ₁₀ aryl or 5- 10-membered heteroaryl, the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, 4-8 membered heterocyclic group, C ₆ -C ₁₀ aryl or 5-10 membered Heteroaryl is optionally substituted ^by R;

R ² is selected from H, halogen, CN, OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, 4-8 membered heterocyclyl, C ₆ -C ₁₀ aryl or 5- 10-membered heteroaryl, the OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, 4-8 membered heterocyclic group, C ₆ -C ₁₀ aryl or 5-10 membered Heteroaryl is optionally substituted ^by R;

Each R ^d is independently selected from halogen, CN, OH, NH ₂ , SH, C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-8 membered heterocyclyl;

Each R ^f is independently selected from H, halogen, CN, OH, NH ₂ , C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl or 4-8 membered heterocyclyl;

n is independently selected from 0, 1 or 2.

In some embodiments, X ² is selected from CR ^2a , CR ^2a R ^2b or NR ^2a .

In some embodiments, X ³ is selected from O, C(=O), CR ^3a R ^3b , or NR ^3a .

In some embodiments, X ³ is selected from C(=O) or CR ^3a R ^3b .

In some embodiments, X ⁴ is selected from O, C(=O), CR ^4a R ^4b , or NR ^4a .

In some embodiments, R ^1a , R ^2a , R ^2b , R ^3a , R ^3b , R ^4a , R ^4b are independently selected from H, halogen, OH, NH ₂ , -C(O) ^Rf , -C( O)N(R ^f ) ₂ , -C(O)OR ^f , -NHC(O)R ^f , -NHC(O)OR ^f , C ₁ -C ₆ alkyl, C ₅ -C ₆ cycloalkyl, C ₆ -C ₁₀ aryl 4-8 membered heterocyclic group or 5-6 membered heteroaryl group, said OH, NH ₂ , C ₁ -C ₆ alkyl, C ₅ -C ₆ cycloalkyl, C ₆ - C ₁₀ aryl 4-8 membered heterocyclic group or 5-6 membered heteroaryl group is optionally substituted by ^Rd .

In some embodiments, R ^1a , R ^2a , R ^2b , R ^3a , R ^3b , R ^4a , R ^4b are independently selected from H, halogen, OH, NH ₂ , -C(O)OR ^f or C ₁ - C ₆ alkyl, the OH, NH ₂ or C ₁ -C ₆ alkyl is optionally substituted by R ^d .

In some embodiments, R ^1a , R ^2a , R ^2b , R ^3a , R ^3b , R ^4a , R ^4b are independently selected from H, halogen, OH, NH ₂ , -C(O)OH, or C ₁ -C ₃ alkyl, the OH, NH ₂ or C ₁ -C ₃ alkyl is optionally substituted by R ^d .

In some embodiments, R ^1a , R ^4a and the cyclic group to which they are attached form a carbon-bridged ring.

In some embodiments, R ^2a , R ^3a and the atoms to which they are attached together form a cycloalkyl group.

In some embodiments, R ^2a , R ^3a and the atoms to which they are attached together form a C ₃ -C ₆ cycloalkyl group.

In some embodiments, R ^3a , R ^3b and the atoms to which they are attached together form a cycloalkyl group.

In some embodiments, R ^3a , R ^3b and the atoms they are connected to together form a C ₃ -C ₆ cycloalkyl group.

In some embodiments, R ¹ is independently _{selected from halogen, CN, OH, NH 2 , or C 1 -C 6} _alkyl _optionally _substituted _with _R ^d .

In some embodiments, R ² is _selected from H, halogen, CN, OH, NH ₂ , C ₁ _-C ₆ alkyl _optionally substituted with R ^d .

In some embodiments, ^R2 is selected from H or halogen.

In some embodiments, R ^is selected from H or F.

In some embodiments, n is independently selected from 0 or 1.

In some embodiments, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from a compound of formula (I') or a pharmaceutically acceptable salt thereof,

Wherein, X ¹ is selected from CR ^1a or N; X ² is selected from CR ^2a R ^2b or NR ^2a ;

R ^1a , R ^2a , R ^2b , X ³ , X ⁴ , Y ¹ , Y ² , Y ³ , R ¹ , n are as defined above.

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

The present disclosure also provides a compound represented by formula (II) and a pharmaceutically acceptable salt thereof:

Among them, [] indicates that L can be connected with X ² , X ³ , X ⁴ , Y ¹ , Y ² or Y ³ ;

X ¹ , X ² , X ³ , X ⁴ , Y ¹ , Y ² , Y ³ , R ¹ , n are as defined above;

When L is connected with Y ¹ , Y ² or Y ³ , Y ¹ , Y ² or Y ³ is C, when L is connected with X ² , X ² is C, CR ^2a or N, when L is connected with X ³ , X ³ is CR ^3a or N, when L is connected with X ⁴ , X ³ is CR ^4a or N;

L is selected from the following structural units:

-(A ^L ) _k -;

^{AL is} independently selected from bond, -O-, -S-, -SO-, -SO ₂ -, -SO ₂ NR ¹⁰ -, -C(R ¹⁰ ) ₂ -, -(CH ₂ ) _q -, - (O-CH ₂ -CH ₂ ) _q -, -NR ¹⁰ -, -NR ¹⁰ C(=O)-, -C(=O)-, -C(=S)-, -OC(=O)- , -OC(O)NR ¹⁰ -, -C(O)ONR ¹⁰ -, -CR ¹⁰ = CR ¹⁰ -, -C≡C-, -P(O)R ¹⁰ -, -P(O)OR ¹⁰ - ,

C ₃ -C ₁₀ cycloalkyl, 4-10 membered heterocyclic group, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl, said C ₃ -C ₁₀ cycloalkyl, 4-10 membered heterocyclic Base, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl are optionally substituted by R ¹¹ ;

R ¹⁰ is selected from H, OH, NH ₂ or C ₁ -C ₆ alkyl;

R ¹¹ is selected from halogen, CN, OH, NH ₂ or C ₁ -C ₆ alkyl;

k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

q is selected from 0, 1, 2, 3, 4, 5 or 6.

In some embodiments, k is selected from 1, 2, 3, 4, 5, 6, 7, 8 or 9.

In some embodiments,

Indicates a single key.

In some embodiments, q is selected from 1, 2, 3, 4, 5 or 6.

In some embodiments, ^AL is independently selected from -O-, -S-, -SO-, -SO ₂ -, -SO ₂ NR ¹⁰ -, -C(R ¹⁰ ) ₂ -, -(CH ₂ ) _q -, -(O-CH ₂ -CH ₂ ) _q -, -NR ¹⁰ -, -NR ¹⁰ C(=O)-, -C(=O)-, -C(=S)-, -OC( ＝O)-, -OC(O)NR ¹⁰ -, -C(O)ONR ¹⁰ -, -CR ¹⁰ ＝CR ¹⁰ -, -C≡C-, -P(O)R ¹⁰ -, -P(O ) OR ¹⁰ -,

C ₃ -C ₁₀ cycloalkyl, 4-10 membered heterocyclic group, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl, said C ₃ -C ₁₀ cycloalkyl, 4-10 membered heterocyclic radical, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl is optionally substituted by R ¹¹ .

In some embodiments, ^AL is independently selected from -O-, -S-, -C(R ¹⁰ ) ₂ -, -(CH ₂ ) _q -, -(O-CH ₂ -CH ₂ ) _q -, -NR ¹⁰ -, -NR ¹⁰ C(=O)-, -C(=O)-, C ₃ -C ₁₀ cycloalkyl, 4-10 membered heterocyclyl, C ₆ -C ₁₀ aryl or 5- 10-membered heteroaryl, the C ₃ -C ₁₀ cycloalkyl, 4-10 membered heterocyclyl, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl is optionally substituted by R ¹¹ .

In some embodiments, ^AL is independently selected from a bond, -O-, -S-, -C(R ¹⁰ ) ₂ -, -(CH ₂ ) _q -, -(O-CH ₂ -CH ₂ ) _q -, -NR ¹⁰ -, -NR ¹⁰ C(=O)-, -C(=O)-, C ₃ -C ₁₀ cycloalkyl, 4-10 membered heterocyclyl, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl, said C ₃ -C ₁₀ cycloalkyl, 4-10 membered heterocyclyl, C ₆ -C ₁₀ aryl or 5-10 membered heteroaryl is optionally substituted by R ¹¹ .

In some embodiments, ^AL is independently selected from -O-, -(CH ₂ ) _q -, -(O-CH ₂ -CH ₂ ) _q -, -NR ¹⁰ -, -NR ¹⁰ C(=O) -, -C(=O)-, C ₃ -C ₁₀ cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl or C ₆ -C ₁₀ aryl, said C ₃ -C ₁₀ Cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl or C ₆ -C ₁₀ aryl is optionally substituted by R ¹¹ .

In some embodiments, ^AL is independently selected from a bond, -O-, -C(R ¹⁰ ) ₂ -, -(CH ₂ ) _q -, -(O-CH ₂ -CH ₂ ) _q -, -NR ¹⁰ -, -NR ¹⁰ C(=O)-, -C(=O)-, 5-6 membered heterocyclic group or 5-6 membered heteroaryl group, said 5-6 membered heterocyclic group or 5-6 The membered heteroaryl is optionally substituted by R ¹¹ .

In some embodiments, ^AL is independently selected from -O-, -(CH ₂ ) _q -, -(O-CH ₂ -CH ₂ ) _q -, -NR ¹⁰ -, -NR ¹⁰ C(=O) -, -C(=O)-, piperazinyl, triazolyl, cyclohexyl, or phenyl optionally substituted by R ¹¹ .

In some embodiments, ^AL is independently selected from a bond, -O-, -C(R ¹⁰ ) ₂ -, -(CH ₂ ) _q -, -(O-CH ₂ -CH ₂ ) _q -, -NR ¹⁰ -, -NR ¹⁰ C(=O)-, -C(=O)-, piperazinyl or triazolyl.

In some embodiments, L is selected from the following structural units:

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(5-10 heteroaryl)-(CH2) _q -C(=O)NH-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -NR ¹⁰ -;

-(O-CH ₂ -CH ₂ ) _q -(5-10 heteroaryl)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-;

-NR ¹⁰ -(CH ₂ ) _q -(5-10heteroaryl)-(CH ₂ ) _q -O-(CH ₂ ) _q -C(=O)-(CH ₂ ) _q -NR ¹⁰ -;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -NHC(=O)-;

-NR ¹⁰ -(CH ₂ ) _q -(5-10 heteroaryl)-(CH ₂ ) _q -NR ¹⁰ -;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NR ¹⁰ -;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-(CH ₂ ) _q -C(=O)-;

-NR ¹⁰ -(CH ₂ ) _q -(5-10heteroaryl)-(CH ₂ ) _q -O-(CH ₂ ) _q -C(=O)-(CH ₂ ) _q -;

-(4-10 membered heterocyclyl)-C(=O)-(CH ₂ ) _q -C(=O)NH-(CH ₂ ) _q -NR ¹⁰ -;

-(5-10 Heteroaryl)-C(=O)-(CH ₂ ) _q -C(=O)NH-(CH ₂ ) _q -NR ¹⁰ -;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -;

-O-(CH ₂ ) _q -(5-10 heteroaryl)-C(=O)-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-;

-O-(CH ₂ ) _q -(CH ₂ ) _q -;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -NR ¹⁰ -;

-O-(CH ₂ ) _q -(CH ₂ ) _q -O-;

-O-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -(CH ₂ ) _q -C(=O)-(CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(4-10 membered heterocyclyl)-(CH ₂ ) _q -NR ¹⁰ -;

-NR ¹⁰ -(CH ₂ ) _q -(4-10 membered heterocyclyl)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(4-10 membered heterocyclyl)-(CH ₂ ) _q -(CH ₂ ) _q -O-;

-OC(=O)-(C ₃ -C ₁₀ cycloalkyl)-(CH ₂ ) _q -O-;

-OC(=O)-(C ₃ -C ₁₀ cycloalkyl)-(CH ₂ ) _q -NH-;

-OC(=O)-(C ₃ -C ₁₀ cycloalkyl)-(CH ₂ ) _q -;

-(CH ₂ ) _q -NHC(=O)-NH-(C ₆ -C ₁₀ aryl)-;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -C(=O)-;

-NR ¹⁰ -(CH ₂ ) _q -C(=O)NH-(CH ₂ ) _q -;

The C ₃ -C ₁₀ cycloalkyl, C ₆ -C ₁₀ aryl, 5-10 heteroaryl or 4-10 membered heterocyclic group is optionally substituted by R ¹¹ , q is as defined above.

In some embodiments, L is selected from the following structural units:

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(5-10 heteroaryl)-(CH2) _q -C(=O)NH-(CH ₂ ) _q -O-;

-(O-CH ₂ -CH ₂ ) _q -(5-10 heteroaryl)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -NHC(=O)-;

-NR ¹⁰ -(CH ₂ ) _q -(5-10 heteroaryl)-(CH ₂ ) _q -NR ¹⁰ -;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NR ¹⁰ -;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-(CH ₂ ) _q -C(=O)-;

-(5-10 Heteroaryl)-C(=O)-(CH ₂ ) _q -C(=O)NH-(CH ₂ ) _q -NR ¹⁰ -;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -;

-O-(CH ₂ ) _q -(5-10 heteroaryl)-C(=O)-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-;

-O-(CH ₂ ) _q -(CH ₂ ) _q -;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(CH ₂ ) _q -NR ¹⁰ -;

-O-(CH ₂ ) _q -(CH ₂ ) _q -O-;

-O-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(4-10 membered heterocyclyl)-(CH ₂ ) _q -NR ¹⁰ -;

-NR ¹⁰ -(CH ₂ ) _q -(4-10 membered heterocyclyl)-(CH ₂ ) _q -O-;

-NR ¹⁰ -(CH ₂ ) _q -(4-10 membered heterocyclyl)-(CH ₂ ) _q -(CH ₂ ) _q -O-.

In some embodiments, L is selected from the following structural units:

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -(CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -triazolyl-(CH ₂ ) _q -C(=O)NH-(CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -NH-;

-(O-CH ₂ -CH ₂ ) _q -triazolyl-(CH ₂ ) _q -O-;

-O-(CH ₂ ) _q -(CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-;

-NH-(CH ₂ ) _q -triazolyl-(CH ₂ ) _q -O-(CH ₂ ) _q -C(=O)-(CH ₂ ) _q -NH-;

-NH-(CH ₂ ) _q -(CH ₂ ) _q -NHC(=O)-;

-O-(CH ₂ ) _q -(CH ₂ ) _q -;

-NH-(CH ₂ ) _q -triazolyl-(CH ₂ ) _q -NH-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NH-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-(CH ₂ ) _q -C(=O)-;

-NH-(CH ₂ ) _q -triazolyl-(CH ₂ ) _q -O-(CH ₂ ) _q -C(=O)-(CH ₂ ) _q -;

-piperazinyl-C(=O)-( _CH2 ) _q -C(=O)NH-( _CH2 ) _q -NH-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -;

-O-(CH ₂ ) _q -triazolyl-C(=O)-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -NH-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-;

-OC(=O)-cyclohexyl-( _CH2 ) _q -O-;

-OC(=O)-cyclohexyl-( _CH2 ) _q -NH-;

-OC(=O)-cyclohexyl-(CH ₂ ) _q -;

-Phenyl-NHC(=O)-NH-(CH ₂ ) _q -;

-NH-(CH ₂ ) _q -(CH ₂ ) _q -C(=O)-;

-NH-(CH ₂ ) _q -C(=O)NH-(CH ₂ ) _q -;

The triazolyl, piperazinyl, cyclohexyl or phenyl is optionally substituted by R ¹¹ , q is as defined above.

In some embodiments, L is selected from the following structural units:

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -(CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -triazolyl-(CH ₂ ) _q -C(=O)NH-(CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -NH-;

-(O-CH ₂ -CH ₂ ) _q -triazolyl-(CH ₂ ) _q -O-;

-O-(CH ₂ ) _q -(CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-;

-NH-(CH ₂ ) _q -(CH ₂ ) _q -NHC(=O)-;

-O-(CH ₂ ) _q -(CH ₂ ) _q -;

-NH-(CH ₂ ) _q -triazolyl-(CH ₂ ) _q -NH-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NH-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-(CH ₂ ) _q -C(=O)-;

-NH-(CH ₂ ) _q -triazolyl-(CH ₂ ) _q -O-(CH ₂ ) _q -C(=O)-(CH ₂ ) _q -;

-piperazinyl-C(=O)-( _CH2 ) _q -C(=O)NH-( _CH2 ) _q -NH-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -O-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -;

-O-(CH ₂ ) _q -triazolyl-C(=O)-;

-NH-(CH ₂ ) _q -(O-CH ₂ -CH ₂ ) _q -NHC(=O)-(CH ₂ ) _q -NH-;

-NH-( _CH2 ) _q- (O- _CH2 - _CH2 ) _q -NHC(=O)-.

In some embodiments, L is selected from the following structural units:

In some embodiments, the compound represented by formula (II) or a pharmaceutically acceptable salt thereof is selected from a compound of formula (II') or a pharmaceutically acceptable salt thereof,

R ^1a , R ^2a , R ^2b , X ³ , X ⁴ , Y ¹ , Y ² , Y ³ , R ¹ , n, L, [] are as defined above.

In some embodiments, the compound represented by formula (II) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

Further, the present disclosure also provides a pharmaceutical composition, which comprises the compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients.

Further, the present disclosure relates to the use of the compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a medicament for preventing or treating diseases mediated by CRBN.

Further, the present disclosure relates to the use of the compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the prevention or treatment of diseases mediated by CRBN.

Further, the present disclosure relates to a compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for preventing or treating CRBN-mediated related diseases.

Further, the present disclosure also relates to a method for treating CRBN-mediated related diseases, the method comprising administering to the patient a therapeutically effective dose of a compound represented by formula (I) or (II) described in the present disclosure or a pharmaceutically acceptable Pharmaceutical preparations of salts.

In another aspect, the present disclosure provides a method for treating mammals suffering from abnormal cell proliferation diseases, comprising administering a therapeutically effective amount of a compound represented by formula (I) or (II) or a pharmaceutically effective amount thereof to a mammal in need of the treatment, preferably a human. acceptable salts, or pharmaceutical compositions thereof.

In another aspect, the present disclosure provides the use of the compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a medicament for preventing or treating abnormal cell proliferation diseases.

In another aspect, the present disclosure provides the use of the compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in preventing or treating abnormal cell proliferation diseases.

In another aspect, the present disclosure provides a compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for preventing or treating abnormal cell proliferation diseases.

In some embodiments, the disorder of abnormal cell proliferation is selected from cancer.

In some embodiments, the cancer is selected from solid tumors, adenocarcinomas, or hematological tumors.

Further, the present disclosure relates to a compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof, as a synthetic intermediate of a bifunctional compound targeting protein degradation, during the preparation of a bifunctional compound targeting protein degradation ( PROTAC molecules).

The present disclosure also provides a compound represented by formula (III) and a pharmaceutically acceptable salt thereof:

Wherein, [], X ¹ , X ² , X ³ , X ⁴ , Y ¹ , Y ² , Y ³ , R ¹ , L, and n are as defined above;

The PTM is a targeting protein binding moiety.

In some embodiments, the compound represented by formula (III) or a pharmaceutically acceptable salt thereof is selected from a compound represented by formula (III') or a pharmaceutically acceptable salt thereof:

R ^1a , R ^2a , R ^2b , X ³ , X ⁴ , Y ¹ , Y ² , Y ³ , R ¹ , n, L, PTM, [] are as defined above.

In some embodiments, the compound represented by formula (III) or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (IIIa) or a pharmaceutically acceptable salt thereof:

Wherein, X ¹ , X ² , X ³ , X ⁴ , Y ¹ , Y ² , Y ³ , R ¹ , n, L, and PTM are as defined above.

In some embodiments, the compound shown in formula (III) or its pharmaceutically acceptable salt is selected from the compound shown in formula (IIIb) or its pharmaceutically acceptable salt:

In some embodiments, the compound represented by formula (III) or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (IIIc) or a pharmaceutically acceptable salt thereof:

In some embodiments, the PTM is selected from the binding portion of the following targeting proteins: ALK, AR, BET1, BRAF, BRCA2, BRD4, BRD9, BTK, CBL, CCNE1, CCNE2, CCR4, CCR7, CCR9, CD47, CLDN18, CYP , DDR1, DMPK, EGFR, ERBB2, ERBB3, ERBB4, FGFR1, FGFR2, FGFR3, FGFR4, GSPT1, JAK1, JAK3, KIF18A, KRAS, LCK, MET, NTRK1, NTRK2, NTRK3, PCSK9, PKMYT1, PARP7, PARP14, RAD51 , RBM10, RET, RORA, STAT3, SOS1, TYK2, USP1 or USP14.

In some embodiments, the PTM is selected from the binding portion of the following targeting proteins: ALK, AR, BET1, BRAF, BRCA2, BRD4, BRD9, BTK, CBL, CCNE1, CCNE2, CCR4, CCR7, CCR9, CD47, CLDN18, CYP , DDR1, DMPK, EGFR, ERBB2, ERBB3, ERBB4, FGFR1, FGFR2, FGFR3, FGFR4, GSPT1, JAK1, JAK3, KIF18A, KRAS, LCK, MET, NTRK1, NTRK2, NTRK3, PCSK9, PKMYT1, PARP7, PARP14, RAD51 , RBM10, RET, RORA, SOS1, TYK2, USP1 or USP14.

In some embodiments, the compound represented by formula (III) 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 also provides a pharmaceutical composition, which comprises the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients.

In another aspect, the present disclosure provides a method for treating mammals suffering from abnormal cell proliferation, comprising administering a therapeutically effective amount of a compound represented by formula (III) or a pharmaceutically acceptable salt thereof to a mammal in need of the treatment, preferably a human , or a pharmaceutical composition thereof.

In another aspect, the present disclosure provides the use of the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a medicament for preventing or treating abnormal cell proliferation diseases.

In another aspect, the present disclosure provides the use of the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in preventing or treating abnormal cell proliferation diseases.

In another aspect, the present disclosure provides a compound represented by formula (III) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for preventing or treating abnormal cell proliferation diseases.

Unless otherwise stated, terms used in this disclosure have the following meanings, definitions of groups and terms recorded in this disclosure, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, examples The definitions of specific compounds in and etc. may be combined and combined with each other arbitrarily. A specific term should not be regarded as indeterminate or unclear if there is no special definition, but should be understood according to the ordinary meaning in the art. When a trade name appears herein, it is intended to refer to its corresponding trade name or its active ingredient.

In this article

Indicates the junction site.

Graphical representations of racemic or enantiomerically pure compounds herein are from Maehr, J. Chem. Ed. 1985, 62:114-120. Unless otherwise specified, with wedge and dotted keys

Indicates the absolute configuration of a stereocenter with black real and virtual bonds

Indicates the relative configuration of a stereocenter (such as the cis-trans configuration of an alicyclic compound).

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

The term "stereoisomer" refers to isomers resulting from differences in the arrangement of atoms in a molecule in space, 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, and thus 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 and Z geometric isomers, (-)- and (+)-enantiomers, (R)- and (S )-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic or other mixtures thereof, such as enantiomers or diastereomers Enriched mixtures, all of the above isomers and mixtures thereof are within the definition of the disclosed compounds. There may be additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms or asymmetric phosphorus atoms in substituents such as alkyl groups, and these isomers and their mixtures involved in all substituents are also included in Compounds of the disclosure are within the definition. Compounds of the present disclosure containing asymmetric atoms can be isolated in optically pure or racemic forms, optically pure forms 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 the specified atom are replaced by a substituent, as long as the valence of the specified atom is normal and the substituted compound is stable. When a substituent is oxo (ie, =0), it means that two hydrogen atoms are replaced, and oxo does not occur on an aromatic group.

The term "optional" or "optionally" means that the subsequently described event or circumstance can or cannot occur, and that description includes that said event or circumstance occurs and that it does not. For example, ethyl is "optionally" substituted with _halogen , meaning that the ethyl group can be unsubstituted ( _CH2CH3 ), monosubstituted ( _CH2CH2F _, _CH2CH2Cl _, etc.), polysubstituted ( _CHFCH2F , _CH2CHF2 , _CHFCH2Cl , _CH2CHCl2 _, _etc. ₎ or fully substituted ( _CF2CF3 , _CF2CCl3 , _CCl2CCl3 _, etc.) _. It will be appreciated by those skilled in the art that for any group containing one or more substituents, no sterically impossible and/or synthetically impossible substitution or substitution pattern is introduced.

When any variable (eg R ^a , R ^b ) occurs more than once in the composition or structure of a compound, its definition is independent at each occurrence. For example, if a group is substituted by 2 R ^b , each R ^b has independent 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 chemical bond or absence, it means that the two groups connected are directly connected. For example, when L in A-L-Z represents a bond, it means that the structure is actually A-Z.

When the linking group mentioned herein does not indicate its linking direction, its linking direction is arbitrary. For example when the structural unit

When L ¹ in is selected from "-C ₁ -C ₃ alkylene-O-", at this time L ¹ can be connected with ring Q and R ¹ from left to right to form "ring QC ₁ -C ₃ Alkylene-OR ¹ ", ring Q and R ¹ can also be linked from right to left to form "ring QOC ₁ -C ₃ alkylene-R ¹ ".

When a bond of a substituent cross-links two atoms in a ring, the substituent may be bonded to any atom on the ring. For example, the structural unit

Indicates that R ¹ can be substituted at any position on the ring, when R ¹ replaces the H of NH, the structure is

_Cm - _Cn herein refers to having an integer number of carbon atoms in the range of mn. For example, "C ₁ -C ₁₀ " means that the group can 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.

In this paper, the bonds depicted by solid and dashed lines

Indicates a single or double bond. For example, the structural unit

Include

The term "alkyl" refers to a hydrocarbon group having the general formula C _n H _2n+1 , and the alkyl group may be straight or branched. The term "C ₁ -C ₁₀ alkyl" is understood to mean a straight-chain or branched saturated 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-di Methylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc.; the term "C ₁ -C ₆ alkyl" can be understood as an alkyl group having 1 to 6 carbon atoms, specific examples 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, etc. The term "C ₁ -C ₃ alkyl" is understood to mean a straight-chain or branched saturated alkyl group having 1, 2 or 3 carbon atoms. The "C ₁ -C ₁₀ alkyl" may include "C ₁ -C ₆ alkyl" or "C ₁ -C ₃ alkyl", etc., and the "C ₁ -C ₆ alkyl" may further include " C ₁ -C ₃ alkyl".

The term "cycloalkyl" refers to a fully saturated carbocyclic ring in the form of a monocyclic ring, a double ring, a bridged ring, or a spiro ring. Unless otherwise indicated, the carbocycle is typically a 3 to 10 membered ring. The term "C ₃ -C ₁₀ cycloalkyl" is understood to mean a saturated monocyclic, fused, spiro or bridged ring having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Specific examples of said 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] decanyl, etc. The term "C ₃ -C ₁₀ cycloalkyl" may include "C ₃ -C ₆ cycloalkyl", and the term "C ₃ -C ₆ cycloalkyl" can be understood as representing a saturated monocyclic or bicyclic hydrocarbon ring, which has 3-6 carbon atoms, specific examples include but not limited to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl and the like.

The term "heterocyclic group" refers to a fully saturated or partially saturated (heteroaromatic not aromatic as a whole) monocyclic, cyclic, spiro or bridged ring group containing 12, 3, 4 or 5 heteroatoms or heteroatom groups (ie, heteroatom-containing atomic groups), said "heteroatoms or heteroatom groups" include but are not limited to nitrogen atoms (N), oxygen atoms (O), sulfur atoms (S), phosphorus atom (P), boron atom (B), -S(=O) ₂ -, -S(=O)-, -P(=O) ₂ -, -P(=O)-, -NH-, - S(=O)(=NH)-, -C(=O)NH- or -NHC(=O)NH-, etc. The term "3-10 membered heterocyclic group" refers to a heterocyclic group with 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms, and its ring atoms contain 1, 2, 3, 4 or 5 independently selected from the heteroatoms or heteroatom groups described above. "3-10 membered heterocyclic group" includes "4-7 membered heterocyclic group", wherein, specific examples of 4-membered heterocyclic group include but are not limited to azetidinyl or oxetanyl; 5-membered Specific examples of heterocyclic groups 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, tetrahydropyridyl or 4H-[1,3,4]thiadiazinyl; specific examples of 7-membered heterocyclyl include, but are not limited to, diazepanyl. The heterocyclic group can also be a bicyclic group, wherein, specific examples of the 5,5-membered bicyclic group include, but are not limited to, hexahydrocyclopenta[c]pyrrol-2(1H)-yl; 5,6-membered bicyclic group Specific examples include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-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 benzofused cyclic group of the above-mentioned 4-7 membered heterocyclic group, specific examples include but not limited to dihydroisoquinolyl and the like. "4-10 membered heterocyclic group" may include "5-10 membered heterocyclic group", "4-7 membered heterocyclic group", "5-6 membered heterocyclic group", "6-8 membered heterocyclic group" , "4-10 membered heterocycloalkyl", "5-10 membered heterocycloalkyl", "4-7 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", "6-8 membered "Heterocycloalkyl" and other ranges, "4-7 membered heterocyclyl" may further include "4-6 membered heterocyclyl", "5-6 membered heterocyclyl", "4-7 membered heterocyclyl" , "4-6 membered heterocycloalkyl", "5-6 membered heterocycloalkyl" and other ranges. Although some bicyclic heterocyclic groups in the present disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclic groups as a whole are non-aromatic.

The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π-electron system. The aryl group can 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 an aryl group having 6 to 20 carbon atoms. Especially rings with 6 carbon atoms (“C _aryl ”), such as phenyl; or rings with ₉ carbon atoms (“C aryl”), such as indanyl or indenyl; or rings with 10 a ring of 3 carbon atoms (“C ₁₀ aryl”), such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl; or a ring of 13 carbon atoms (“C ₁₃ aryl”), such as fluorenyl; or is a ring having 14 carbon atoms ("C ₁₄ aryl"), such as anthracenyl. The term "C ₆ -C ₁₀ aryl" is understood to mean an aryl group having 6 to 10 carbon atoms. Especially rings with 6 carbon atoms (“C _aryl ”), such as phenyl; or rings with ₉ carbon atoms (“C aryl”), such as indanyl or indenyl; or rings with 10 carbon atoms (“C ₁₀ aryl”), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl. The term "C ₆ -C ₂₀ aryl" may include "C ₆ -C ₁₀ aryl"

The term "heteroaryl" refers to an aromatic monocyclic or fused polycyclic ring system, which contains at least one ring atom selected from N, O, and S, and an aromatic ring group whose ring atoms are C. The term "5-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms, and it contains, 2, 3, 4 or 5, preferably 1, 2 or 3 heteroatoms independently selected from N, O and S. In particular, heteroaryl is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiazolyl Diazolyl, etc. and their benzo derivatives, such as benzofuryl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazole base, indazolyl, indolyl or isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, or triazinyl, etc., and their benzo derivatives, such as quinolinyl, quinazole Linyl or isoquinolinyl, etc.; or azocinyl, indolizinyl, purinyl, etc. and their benzo derivatives; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthalene Pyridyl, pteridyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, etc. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and which contains 1, 2 or 3, preferably 1-2, heteroatoms independently selected from N, O and S .

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

The term "hydroxyl" refers to a -OH group.

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

The term "mercapto" refers to a -SH group.

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

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

The term "therapeutically effective amount" means (i) treating a particular disease, condition or disorder, (ii) alleviating, ameliorating or eliminating one or more symptoms of a particular disease, condition or disorder, or (iii) delaying The amount of a compound of the disclosure required for the onset of one or more symptoms of a particular disease, condition or disorder. The amount of a compound of the present disclosure that constitutes a "therapeutically effective amount" will vary 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 according to its own knowledge and this 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 human and animal tissues without excessive Toxicity, irritation, allergic reaction, or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to salts of pharmaceutically acceptable acids or bases, including salts formed between compounds and inorganic or organic acids, and salts formed between compounds and inorganic or organic bases.

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

The term "pharmaceutically acceptable excipients" refers to those excipients that have no obvious stimulating effect on the organism and will 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 and the like.

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

The disclosure also includes isotopically labeled compounds of the disclosure that are identical to those described herein, but with one or more atoms replaced by an atom of 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.

Certain isotopically labeled compounds of the disclosure (eg, ^with3H ^and14C ) are useful in compound and/or substrate tissue distribution assays. Tritiated ( ^ie3H ) and carbon-14 ( ^ie14C ) isotopes are especially 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 following procedures similar to those disclosed in the Schemes and/or Examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

The pharmaceutical composition of the present disclosure can be prepared by combining the compound of the present disclosure with suitable pharmaceutically acceptable auxiliary materials, for example, it 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, etc.

Typical routes of administration of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition 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 produced by methods well known in the art, such as conventional mixing methods, dissolving methods, granulating methods, emulsifying methods, freeze-drying methods and the like.

In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical compositions can be formulated by mixing the active compounds 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 and the like for oral administration to patients.

Solid oral compositions can be prepared by conventional methods of mixing, filling or tabletting. It can be obtained, for example, by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if desired, and processing the mixture into granules to obtain tablets Or the core of the sugar coating. Suitable auxiliary materials include but are not limited to: binders, diluents, disintegrants, lubricants, glidants or flavoring agents, etc.

The pharmaceutical composition may also be adapted for parenteral administration as a suitable unit dosage form of sterile solutions, suspensions or lyophilized products.

In all methods of administration of the compound of general formula I described herein, the daily dosage is 0.01 mg/kg to 200 mg/kg body weight, preferably 0.05 mg/kg to 50 mg/kg body weight, more preferably 0.1 mg/kg to 30 mg /kg body weight, in single or divided doses.

Detailed ways

The invention will be described in detail through the examples below, but it does not imply any unfavorable limitation to the present disclosure. The present disclosure has been described in detail herein, and its specific embodiments are also disclosed. For those skilled in the art, various changes and improvements can be made to the specific embodiments of the present disclosure without departing from the spirit and scope of the present disclosure. will be obvious. All reagents used in this disclosure are commercially available and used without further purification.

Unless otherwise specified, the ratios indicated for mixed solvents are volume mixing ratios.

Unless otherwise stated, % means wt%.

compound by hand or

The software is named, and the commercially available compounds adopt the supplier catalog name.

Compound structures were determined by nuclear magnetic resonance (NMR) and/or mass spectroscopy (MS). The unit of NMR shift is 10 ^-6 (ppm). The solvents measured by NMR are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is _{tetramethylsilane} (TMS); concentration.

The preparation method of the compound represented by the formula (f) of the present disclosure or a pharmaceutically acceptable salt thereof is as follows:

The preparation method of the compound represented by the present disclosure formula (f') or a pharmaceutically acceptable salt thereof is as follows:

Example 1: 3-(2-Carbonyl-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione

Step 1: 6,7,8,8a-Tetrahydrobenzo[cd]indol-2(1H)-one (1-2)

Dissolve benzo[cd]indol-2(1H)-one (2.0g, 11.8mmol) in trifluoroacetic acid (10mL), add palladium carbon (200mg, 10%wt), replace hydrogen three times, and heat to 50°C , reacted overnight, cooled to room temperature, filtered, concentrated to dryness, and reversed-phase purified to obtain 6,7,8,8a-tetrahydrobenzo[cd]indol-2(1H)-one (950mg, yield 50%, purity 96%). m/z (ESI): 174[M+H] ⁺ .

Step 2: 2,6-Bis(benzyloxy)pyridine (1-4)

Benzyl alcohol (1.1g, 10.2mmol) was added into N,N-dimethylformamide (10mL) at room temperature, and then sodium hydride (680mg, 17mmol) was added to react at room temperature for 1 hour. Compound 2,6-dichloropyridine (0.5 g, 3.4 mmol) was added and reacted overnight at room temperature. The reaction solution was added to ice water (30 mL). The resulting mixture was extracted with ethyl acetate (20mL*3), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and the solvent was distilled off under reduced pressure to obtain 2,6-bis(benzyl oxy)pyridine (960 mg). m/z(ESI):292[M+H] ⁺ .

Step 3: 2,6-Bis(benzyloxy)-3-iodopyridine (1-5)

At room temperature, 2,6-bis(benzyloxy)pyridine (960mg, 3.3mmol) and 1-iodopyrrolidine-2,5-dione (900mg g, 4.0mmol) were added to acetonitrile (10ml), and the resulting mixture The reaction was stirred and heated at 80°C for 8h. The reaction solution was cooled to room temperature, concentrated to dryness and purified by reverse-phase column chromatography (eluent: water: acetonitrile = 1:1) to obtain 2,6-bis(benzyloxy)-3-iodopyridine (550 mg, two steps Yield 39%, purity 95%). m/z (ESI): 418[M+H] ⁺ .

Step 4: 1-(2,6-bis(benzyloxy)pyridin-3-yl)-6,7,8,8a-tetrahydrobenzo[cd]indol-2(1H)-one (1- 6)

Add 6,7,8,8a-tetrahydrobenzo[cd]indol-2(1H)-one (10 mg, 57.8 μmol), 2,6-bis(benzyloxy)-3- Iodopyridine (36 mg, 86.7 μmol), copper iodide 2 (2.2 mg, 11.6 μmol) and potassium carbonate (24 mg, 173.4 μmol) were dissolved in DMF (1 mL), and the ligand trans-N,N'-dimethyl Diamino-1,2-cyclohexanediamine (3.3 mg, 23.2 μmol), vacuumed and replaced argon three times, heated to 100 ° C, reacted for 30 min, cooled to room temperature, reversed-phase column chromatography (water: acetonitrile = 1:1 ) was purified to obtain 1-(2,6-bis(benzyloxy)pyridin-3-yl)-6,7,8,8a-tetrahydrobenzo[cd]indol-2(1H)-one (21mg, Yield 79%). LC-MS: m/z (ESI): 463 [M+H] ⁺ .

Step 5: 3-(2-Oxo-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione (1)

1-(2,6-bis(benzyloxy)pyridin-3-yl)-6,7,8,8a-tetrahydrobenzo[cd]indol-2(1H)-one (21mg, 45.4umol ) was added into ethanol (5mL), palladium carbon (20mg, 100%wt) was added, vacuum was changed for three times for hydrogen, room temperature was reacted overnight, pad diatomaceous earth was filtered, and the filtrate was concentrated and dried by reverse phase column chromatography (eluent was Water: acetonitrile = 1:1) to obtain 3-(2-carbonyl-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)piperidine-2,6-di Ketone (12 mg, yield 93%). m/z(ESI):285[M+H] ⁺ .

¹ H NMR (300MHz, DMSO) δ10.94(s, 1H), 7.43(dd, J=16.3, 7.5Hz, 3H), 5.03–4.77(m, 1H), 4.43(dd, J=11.2, 3.7Hz ,1H),3.07–2.55(m,6H),2.44–2.25(m,1H),2.08–1.88(m,3H).

Example 2: 2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]tri Azolo[4,3-a][1,4]diazoheptin-6-yl)-N-(11-(4-(2,6-dioxopiperidin-3-yl)-5 -Oxo-3,3a,4,5-tetrahydropyrrolo[2,3,4-de]quinolin-1(2H)-yl)-11-carbonylundecyl)acetamide (compound 2)

Step 1: tert-butyl 3-(3-bromoanilino)propionate (2-2)

Compound 2-1 (30g, 174mmol), tert-butyl acrylate (22.4g, 174mmol) and acetic acid (2.09g, 34.9mmol) were added to a 250mL reaction flask, and stirred at 90°C for 12h. After the reaction, the reaction solution was concentrated to obtain crude compound 2-2 (50 g), which was directly used in the next reaction. m/z (ESI): 300[M+H] ⁺ .

Step 2: tert-butyl 3-((N-(3-bromophenyl)-4-methylphenyl)sulfonylamino)propionate (2-3)

Compound 2-2 (20 g, 66.6 mmol) was dissolved in pyridine (100 mL), and p-toluenesulfonyl chloride (25.4 g, 133 mmol) was slowly added dropwise at 0°C. The reaction was stirred at 25 °C for 2 h. After the reaction, slowly add saturated ammonium chloride aqueous solution (100mL) dropwise, use ethyl acetate (50mL*3) to extract the reaction solution, collect the organic phase, wash with saturated ammonium chloride aqueous solution (50mL), dry the organic phase over anhydrous sodium sulfate, After filtration, the filtrate was concentrated to obtain a crude product, which was subjected to column chromatography (petroleum ether: ethyl acetate = 10:1 as eluent) to obtain compound 2-3 (15 g, yield 50%). m/z (ESI): 454[M+H] ⁺ .

Step 3: 3-((N-(3-bromophenyl)-4-methylphenyl)sulfonylamino)propanoic acid (2-4)

Compound 2-3 (15g, 33.0mmol) was dissolved in a mixed solution of trifluoroacetic acid (15mL) and dichloromethane (30mL), and stirred at 25°C for 12h. The reaction solution was concentrated to obtain crude product 2-4 (14 g), which was directly used in the next reaction. m/z(ESI):398[M+H] ⁺ .

Step 4: 5-Bromo-2,3-dihydroquinolin-4(1H)-one (2-5)

Polyphosphoric acid (75g, 191mmol) was heated to 80°C, compound 2-4 (15g, 36.9mmol) was added, the temperature was raised to 110°C, and the reaction was carried out for 0.5h. After the reaction, pour the reaction solution into water (150mL), extract the reaction solution with ethyl acetate (80mL*3), collect the organic phase, wash with saturated ammonium chloride aqueous solution (50mL), and dry the organic phase with anhydrous sodium sulfate , filtered, and the filtrate was concentrated to obtain a crude product, and then subjected to column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain compound 2-5 (4.2 g, yield 25%). m/z(ESI):226[M+H] ⁺ .

Step 5: tert-butyl 5-bromo-4-oxo-3,4-dihydroquinoline-1(2H)-carboxylate (2-6)

Compound 2-5 (4.2g, 18.5mmol) was dissolved in dichloromethane (40mL), then added 4-dimethylaminopyridine (113mg, 0.9mmol), diisopropylethylamine (3.6g, 27.9mmol) and di-tert-butyl dicarbonate (4.05 g, 18.6 mmol). The reaction solution was heated to 50°C and refluxed for 16h. After the reaction, the reaction solution was concentrated to obtain a crude product, which was subjected to column chromatography (eluent: petroleum ether: ethyl acetate = 5:1) to obtain compound 2-6 (2.32 g, yield 77%). m/z(ESI):326[M+H] ⁺ .

Step 6: tert-butyl 4-amino-5-bromo-3,4-dihydroquinoline-1(2H)-carboxylate (2-7)

Dissolve compound 2-6 (2.1g, 6.44mmol) and ammonium acetate (6.6g, 85.6mmol) in methanol (20mL), react at 60°C for 6h, add sodium cyanoborohydride (2.43g, 38.6mmol) , Reacted at 60°C for 10h. After the reaction, slowly add saturated ammonium chloride aqueous solution (50mL) to the reaction solution, extract the reaction solution with ethyl acetate (80mL*3), collect the organic phase, wash with saturated ammonium chloride aqueous solution (50mL), anhydrous sulfuric acid The organic phase was dried with sodium, filtered, and the filtrate was concentrated to obtain a crude product, and then subjected to column chromatography (eluent: petroleum ether: ethyl acetate = 5:1, plus 0.5% (volume ratio) triethylamine) to obtain compound 2-7 (0.75g, yield 29%). m/z(ESI):327[M+H] ⁺ .

Step 7: tert-butyl 5-oxo-3,3a,4,5-tetrahydropyrrolo[2,3,4-de]quinoline-1(2H)-carboxylate (2-8)

Compound 2-7 (0.75g, 2.2mmol), 1,1'-bis(diphenylphosphino)ferrocene (211mg, 0.38mmol), triethylamine (577mg, 5.70mmol), methanol (0.3mL) and palladium acetate (42.7mg, 0.19mmol) were sequentially added to DMF (3mL), carbon monoxide was replaced three times, and heated to 80°C under carbon monoxide (15psi) for 12h. After the reaction, add water (10mL) to the reaction solution, extract the reaction solution with ethyl acetate (20mL*3), collect the organic phase, wash with saturated ammonium chloride aqueous solution (30mL), dry the organic phase with anhydrous sodium sulfate, filter , the filtrate was concentrated to obtain a crude product, and then subjected to column chromatography (eluent: petroleum ether: ethyl acetate = 1:2) to obtain compound 2-8 (0.42 g, yield 75%). m/z(ESI):275[M+H] ⁺ .

Step 8: 4-(2,6-Bis(benzyloxy)pyridin-3-yl)-5-oxo-3,3a,4,5-tetrahydropyrrolo[2,3,4-de]quinone Phenyl-1(2H)-tert-butyl carboxylate (2-9)

Under nitrogen, compound 2-8 (150mg, 0.54mmol), cuprous iodide (34.7mg, 0.11mmol) and compound 1-5 (342mg, 0.82mmol) were dissolved in 1,4-dioxane (2mL ), react at 100°C for 12h. After the reaction was completed, the crude product was concentrated to obtain compound 2-9 (120 mg, yield 39%) by column chromatography (eluent: methanol: dichloromethane = 1:20). m/z(ESI):564[M+H] ⁺ .

Step 9: 4-(2,6-Dicarbonylpiperidin-3-yl)-5-oxo-3,3a,4,5-tetrahydropyrrolo[2,3,4-de]quinoline-1 (2H)-tert-butyl carboxylate (2-10)

Compound 2-9 (150mg, 0.26mmol) was dissolved in ethanol (10mL), palladium/carbon (16mg) was added, hydrogen was replaced 3 times, and reacted at 25°C for 16h under hydrogen (15psi). After the reaction, the crude product compound 2-10 (90mg) was obtained by filtration, which was directly used in the next reaction. m/z(ESI):386[M+H] ⁺ .

Step 10: 3-(5-Oxo-2,3,3a,5-tetrahydropyrrolo[2,3,4-de]quinolin-4(1H)-yl)piperidine-2,6-di Ketones (2-11)

Under nitrogen, compound 2-10 (60mg, 0.15mmol) was dissolved in TFA (1mL) and DCM (5mL) to obtain a mixed solution, and reacted at 25°C for 1h. After the reaction, the crude product was concentrated and purified by reverse-phase column chromatography (eluent: water: acetonitrile = 1:1) to obtain compound 2-11 (38 mg, yield 85%). m/z(ESI):286[M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ10.93 (br s, 1H), 7.14 (dt, J=2.4, 7.6Hz, 1H), 6.81–1 6.74 (m, 1H), 6.63–6.55 (m ,1H),6.25–6.13(m,1H),5.05–4.78(m,1H),4.54–4.38(m,1H),3.54–3.44(m,1H),2.93–2.78(m,1H),2.60 (br d,J=1.3Hz,2H),2.43–2.36(m,1H),2.32–2.10(m,1H),2.06-1.95(m,1H),1.22–1.02(m,1H).

Step 11: (11-(4-(2,6-dioxopiperidin-3-yl)-5-oxo-3,3a,4,5-tetrahydropyrrolo[2,3,4-de ]quinolin-1(2H)-yl)-11-carbonylundecyl)carbamate tert-butyl ester (2-12)

Dissolve 11-(tert-butoxycarbonylamino)undecanoic acid (10mg, 0.03mmol) in thionyl chloride (1mL), heat to 70°C for 10min and concentrate, then add dichloromethane (2mL). Compound 2-11 (1 mg, 0.03 mmol) was added to the above reaction solution, and then triethylamine (1 mg, 0.09 mmol) was added. React at 25°C for 1h. After the reaction was completed, the crude product compound 2-12 (17 mg) was obtained by concentration, which was directly used in the next reaction. m/z(ESI):569[M+H] ⁺ .

Step 12: 3-(1-(11-aminoundecanoyl)-5-oxo-2,3,3a,5-tetrahydropyrrolo[2,3,4-de]quinoline-4(1H )-yl)piperidine-2,6-dione (2-13)

Compound 2-12 (17mg, 0.03mmol) was dissolved in dichloromethane (2mL), then hydrogen chloride/1,4-dioxane solution (4M, 1mL) was added, and reacted at 25°C for 1h. After the reaction was completed, the crude product compound 2-13 (15 mg, hydrochloride) was obtained by concentration, which was directly used in the next reaction. m/z (ESI): 469[M+H] ⁺ .

Step 13: 2-((S)-4-(4-Chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazole And[4,3-a][1,4]diazoheptin-6-yl)-N-(11-(4-(2,6-dioxopiperidin-3-yl)-5- Oxo-3,3a,4,5-tetrahydropyrrolo[2,3,4-de]quinolin-1(2H)-yl)-11-carbonylundecyl)acetamide (Compound 2)

Compound 2-13 (10mg, 0.02mmol), (S)-2-[2,3,9-trimethyl-4-(4-chlorophenyl)-6H-thieno[3,2-f] [1,2,4]Triazolo[4,3-a][1,4]diazepine-6-yl]acetic acid (9mg, 0.02mmol) and 2-(7-azobenzotriazole Azole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (8mg, 0.02mmol) was dissolved in N,N-dimethylformamide (2mL), and diisopropylethylamine was added (8mg, 0.06mmol), react at 25°C for 1h. After the reaction, compound 2 (4 mg, yield 24%) was purified by reverse phase column chromatography (eluent: water: acetonitrile = 1: 1). m/z(ESI):851[M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ11.02(s, 1H), 8.49(s, 1H), 8.19(t, J=5.6Hz, 1H), 7.57–7.37(m, 7H), 5.01( ddd, J＝51.6, 12.9, 4.9Hz, 1H), 4.58–4.50(m, 2H), 3.27(dd, J＝14.9, 8.4Hz, 2H), 3.22–3.04(m, 4H), 2.97–2.83( m,1H),2.70(td,J＝4.5,3.7,2.6Hz,1H),2.66–2.58(m,7H),2.45-2.44(m,1H),2.43(s,3H),2.36-2.34( m,1H),2.09-2.01(m,1H),1.65(s,3H),1.37–1.20(m,14H).

Example 3: 1-(3-chloro-4-methylphenyl)-3-((1-(2,6-dioxopiperidin-3-yl)-2-oxo-1,2, 6,7,8,8a-Hexahydrobenzo[cd]indol-4-yl)methyl)urea

Step 1: 4-Bromo-6,7,8,8a-tetrahydrobenzo[cd]indol-2(1H)-one (3-1)

Silver nitrate (980mg, 5.77mmol) was dissolved in water (4mL) and added to a solution of compound 1-2 (1g, 5.77mmol) in acetic acid (8mL) solution, nitric acid (363mg, 5.77mmol) and liquid bromine (6.46g, 40.41mmol) was added to the reaction solution and reacted at 25°C for 16h. After the reaction, add saturated sodium sulfite aqueous solution dropwise to wash until the solution is clear, use ethyl acetate (20mL*3) to extract the reaction solution, collect the organic phase, wash with saturated ammonium chloride aqueous solution (30mL), dry the organic phase with anhydrous sodium sulfate, filter , the filtrate was concentrated to obtain a crude product, which was then purified by reverse-phase column chromatography (eluent: water: acetonitrile = 1:1) to obtain compound 3-1 (0.5 g, yield 34%). m/z(ESI):252[M+H] ⁺ .

Step 2: Dimethyl 2-(4-bromo-2-oxo-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)glutarate (3-2 )

Dissolve compound 3-1 (100mg, 0.4mmol) in N,N-dimethylformamide (2mL), add sodium hydride (20mg, 0.5mmol) and stir at 25°C for 0.5h, add 2-bromoglutaric acid After dimethyl ester (140mg, 0.6mmol), react at 25°C for 1h. After the reaction, compound 3-2 (100 mg, yield 61%) was purified by reverse phase column chromatography (eluent: water: acetonitrile = 1:1). m/z (ESI): 410[M+H] ⁺ .

Step 3: 2-(4-Bromo-2-oxo-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)glutaric acid (3-3)

Compound 3-2 (50mg, 0.12mmol) was dissolved in tetrahydrofuran (2mL) and water (2mL), lithium hydroxide (9mg, 0.36mmol) was added, and reacted at 25°C for 2h. After the reaction, the pH was adjusted to about 3 with concentrated hydrochloric acid, and purified by reverse phase column chromatography (eluent: water: acetonitrile = 1:1) to obtain compound 3-3 (40 mg, yield 85%). m/z (ESI): 382[M+H] ⁺ .

Step 4: 3-(4-Bromo-2-oxo-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione ( 3-4)

Under nitrogen, compound 3-3 (40mg, 0.1mmol), triethylamine (32mg, 0.3mmol), 1-hydroxybenzotriazole (32mg, 0.24mmol) were dissolved in dichloromethane (2mL), and added Trifluoroacetamide (12mg, 0.1mmol), after the temperature of the reaction solution dropped to 0°C, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (44mg, 0.23mmol) was added , Reacted at 25°C for 16h. After the reaction, compound 3-4 (30 mg, yield 82%) was purified by reverse phase column chromatography (eluent: water: acetonitrile = 1:1). m/z(ESI):363[M+H] ⁺ .

Step 5: ((1-(2,6-dioxopiperidin-3-yl)-2-oxo-1,2,6,7,8,8a-hexahydrobenzo[cd]indole- 4-yl) methyl) tert-butyl carbamate (3-5)

Under nitrogen, compound 3-4 (50mg, 0.13mmol), [(tert-butoxycarbonylamino)methyl] potassium trifluoroborate (65mg, 0.27mmol), palladium acetate (6mg, 0.027mmol), n-butyl Bis(1-adamantyl)phosphine (20mg, 0.55mmol) and cesium carbonate (134mg, 0.41mmol) were dissolved in 1,4-dioxane (2mL) and water (0.2mL), and reacted at 100°C for 1h . After the reaction, compound 3-5 (40 mg, yield 70%) was purified by reverse phase column chromatography (eluent: water: acetonitrile = 1: 1). m/z (ESI): 414[M+H] ⁺ .

Step 6: 3-(4-(Aminomethyl)-2-oxo-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)piperidine-2,6 - diketone (3-6)

Compound 3-5 (40mg) was dissolved in dichloromethane (2mL), hydrogen chloride/1,4-dioxane solution (4M, 1mL) was added, and reacted at 25°C for 1h. After the reaction, it was concentrated to obtain the crude product compound 3-6 (33 mg), which was directly used in the next reaction. m/z (ESI): 314[M+H] ⁺ .

Step 7: 1-(3-Chloro-4-methylphenyl)-3-((1-(2,6-dioxopiperidin-3-yl)-2-oxo-1,2,6 ,7,8,8a-Hexahydrobenzo[cd]indol-4-yl)methyl)urea (3)

Dissolve compound 3-6 (10mg, 0.028mmol), triethylamine (9mg, 0.085mmol) in N,N-dimethylformamide (2mL), add 3-chloro-4-methylbenzene isocyanate (6mg ,0.037mmol), reacted at 25°C for 1h. After the reaction, compound 3 (5 mg, yield 36%) was purified by reverse phase column chromatography (eluent: water: acetonitrile = 1:1). m/z (ESI): 481[M+H] ⁺ .

¹ H NMR (600MHz, DMSO-d ₆ )δ10.95(s, 1H), 8.74(s, 1H), 7.66(d, J=2.3Hz, 1H), 7.49(dd, J=12.1, 7.7Hz, 1H),7.39(dd,J=8.3,3.3Hz,1H),7.18(d,J=8.3Hz,1H),7.13(d,J=8.4Hz,1H),6.84–6.65(m,1H), 4.91(ddd, J=99.9, 13.3, 4.9Hz, 1H), 4.44(dd, J=12.0, 4.5Hz, 1H), 4.36(dd, J=15.7, 5.9Hz, 1H), 4.28(dt, J= 16.2,4.5Hz,1H),2.96(dt,J=17.1,8.2Hz,1H),2.84(qd,J=15.7,13.7,9.6Hz,1H),2.70(dt,J=18.0,8.9Hz,1H ),2.62–2.56(m,2H),2.46–2.28(m,1H),2.23(s,3H)2.17-2.10(m,1H),2.06–1.90(m,2H),1.15-1.04(m, 1H).

Synthetic method 2 of compound 3-4:

Step 1: N-(6-bromo-1,2,3,4-tetrahydronaphthalen-1-yl)picolinamide (3-c)

Compound 3-b (3g, 13.27mmol), 2-pyridinecarboxylic acid (1.96g, 15.92mmol), 1-hydroxybenzotriazole (2.15g, 15.92mmol), 1-(3-dimethylaminopropyl) -3-Ethylcarbodiimide hydrochloride (3.05g, 15.92mmol) was dissolved in N,N-dimethylformamide (20mL), N,N-diisopropylethylamine (4.29g, 33.17mmol), stirred and reacted at 25°C for 16 hours. After the reaction, the reaction liquid was concentrated, and compound 3-c (4 g, yield 91%) was obtained by column chromatography (petroleum ether: ethyl acetate = 10:1). m/z (ESI): 331 [M+H]+.

Step 2: 4-Bromo-6,7,8,8a-tetrahydrobenzo[cd]indol-2(1H)-one (3-1)

Compound 3-c (200mg, 0.6mmol), cobalt acetate tetrahydrate (150mg, 0.6mmol), silver carbonate (499mg, 1.81mmol), pivalic acid (185mg, 1.81mmol), diethyl azodicarboxylate ( 420mg, 2.42mmol) was dissolved in trifluoroethanol (1.5mL) and placed in a polytetrafluoroethylene sealed tube. The reaction solution was stirred and reacted at 130° C. for 16 hours. After the reaction, the reaction solution was filtered, and the filtrate was purified by reverse-phase column chromatography (eluent: water: acetonitrile = 1:1) to obtain compound 3-1 (40 mg, yield 26%). m/z(ESI):252[M+H]+.

Step 3: Dimethyl 2-(4-bromo-2-oxo-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)glutarate (3-2 )

Dissolve compound 3-1 (100mg, 0.4mmol) in N,N-dimethylformamide (2mL), add sodium hydride (20mg, 0.5mmol) and stir at 25°C for 0.5h, add 2-bromopentanedi Dimethyl ester (140mg, 0.6mmol) was reacted at 25°C for 1h. After the reaction, compound 3-2 (100 mg, yield 61%) was purified by reverse phase column chromatography (eluent: water: acetonitrile = 1:1). m/z (ESI): 410[M+H] ⁺ .

Step 4: 2-(4-Bromo-2-oxo-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)glutaric acid (3-3)

Step 5: 3-(4-Bromo-2-oxo-6,7,8,8a-tetrahydrobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione ( 3-4)

Example 4: 6-(2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2, 4] Triazolo[4,3-a][1,4]diazoheptin-6-yl)acetamido)-N-((1-(2,6-dioxopiperidine-3- Base)-2-oxo-1,2,6,7,8,8a-hexahydrobenzo[cd]indol-4-yl)methyl)caproamide

Step 1: (6-(((1-(2,6-dioxopiperidin-3-yl)-2-oxo-1,2,6,7,8,8a-hexahydrobenzo[cd ]indol-4-yl)methyl)amino)-6-carbonylhexyl)carbamate tert-butyl ester (4-1)

Compound 3-6 (33mg, 0.094mmol), tert-butoxycarbonyl 6-aminocaproic acid (22mg, 0.094mmol) and 2-(7-azobenzotriazole)-N,N,N', N'-Tetramethyluronium hexafluorophosphate (39mg, 0.1mmol) was dissolved in N,N-dimethylformamide (2mL), diisopropylethylamine (36mg, 0.28mmol) was added, and at 25°C Reaction 1h. After the reaction, compound 4-1 (40 mg, yield 80%) was purified by reverse phase column chromatography (eluent: water: acetonitrile = 1: 1). m/z(ESI):527[M+H] ⁺ .

Step 2: 6-Amino-N-((1-(2,6-dioxopiperidin-3-yl)-2-oxo-1,2,6,7,8,8a-hexahydrobenzo [cd]indol-4-yl)methyl)hexanamide (4-2)

Compound 4-1 (40mg) was dissolved in dichloromethane (2mL), hydrogen chloride/1,4-dioxane solution (4M, 1mL) was added, and reacted at 25°C for 1h. After the reaction, it was concentrated to obtain the crude product compound 4-2 (30 mg), which was directly used in the next reaction. m/z (ESI): 427[M+H] ⁺ .

Step 3: 6-(2-((S)-4-(4-Chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4 ]triazolo[4,3-a][1,4]diazoheptin-6-yl)acetamido)-N-((1-(2,6-dioxopiperidin-3-yl )-2-oxo-1,2,6,7,8,8a-hexahydrobenzo[cd]indol-4-yl)methyl)caproamide (4)

Compound 4-2 (30mg, 0.064mmol), (S)-2-[2,3,9-trimethyl-4-(4-chlorophenyl)-6H-thieno[3,2-f] [1,2,4]triazolo[4,3-a][1,4]diazepine-6-yl]acetic acid (26mg, 0.064mmol) and 2-(7-azobenzotriazole Azole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (29mg, 0.077mmol) was dissolved in N,N-dimethylformamide (2mL), and diisopropylethylamine was added (25mg, 0.19mmol), react at 25°C for 1h. After the reaction, compound 4 (3 mg, yield 5%) was purified by reverse phase column chromatography (eluent: water: acetonitrile = 1:1). m/z (ESI): 809[M+H] ⁺ .

¹ H NMR (600MHz, DMSO-d ₆ ) δ10.95(d, J=7.0Hz, 1H), 8.29(t, J=6.0Hz, 1H), 8.19(t, J=5.7Hz, 1H), 7.49 (d,J=8.2Hz,2H),7.47–7.44(m,1H),7.42(d,J=8.1Hz,2H),7.31(dd,J=7.8,4.2Hz,1H),5.06–4.79( m,1H),4.51(t,J=7.1Hz,1H),4.42(dd,J=11.9,4.5Hz,1H),4.33(dt,J=16.0,5.1Hz,1H),4.24(dt,J ＝15.4,4.8Hz,1H),3.27-3.23(m,1H),3.20-3.17(m,1H),3.15-3.09(m,1H),2.86-2.80(m,1H),2.70–2.55(m ,7H),2.41(s,3H),2.17(t,J＝7.5Hz,2H),2.13-2.05(m,1H),2.03-1.97(m,3H),1.93-1.91(m,1H), 1.62(s,3H),1.56(t,J=7.5Hz,2H),1.48-1.43(m,2H),1.31(t,J=7.8Hz,2H),1.07(dd,J=17.1,12.1Hz ,1H).

Compounds other than those synthesized in Examples 1-4 can be synthesized by referring to the synthesis routes and source materials in Examples 1-4.

Biological activity and related properties test cases

Test example 1, Cereblon binding experiment

1. Experimental instruments and materials

仪器名称equipment name	设备厂家Equipment manufacturers	型号model
振荡器oscillator	BoxunBoxun	BSD-YX3400BSD-YX3400
读板仪Plate reader	PerkinElmerPerkinElmer	EnvisionEnvision
离心机centrifuge	EppendorfEppendorf	Eppendorf MixmateEppendorf Mixmate
化合物稀释及加样仪Compound Dilution and Sampler	PerkinElmerPerkinElmer	EchoEcho

The detection kit (HTRF Human Cereblon Binding Kits) used in the experiment is a kind of

Assay method for quantitative measurement of Cereblon WT ligand. The detection principle is based on HTRF technology. The specific labeled GST antibody (Euroum Cryptate, donor) is simultaneously combined with GST-labeled human Cereblon WT ligand and XL665-labeled lenalidomide tracer (acceptor), and excited by light source The donor triggers fluorescence resonance energy transfer (FRET) to the acceptor, and the acceptor emits fluorescence at a specific wavelength of 665nm. After adding the compound, it competes with XL665-labeled lenalidomide to prevent FRET from occurring. The FRET signal ratio is inversely proportional to the compound concentration.

Other reagents and consumables required for the experiment are as follows:

试剂Reagent	品牌brand	货号Item No.
HTRF Human Cereblon Binding KitsHTRF Human Cereblon Binding Kits	PromegaPromega	64BDCRBNPEG64BDCRBNPEG
384孔板384-well plate	PerkinElmerPerkinElmer	60072996007299

2. Experimental steps

The disclosed compound was dissolved in DMSO, and the stock concentration of the stock solution was 10 mM. Gradient dilution of the compound mother solution is carried out through the compound dilution and sample loading program of the Echo instrument. The total experimental system of the dilution program is 20 μL, the initial concentration of the compound to be tested is 100 μM, the initial concentration of the standard is 200 μM, 4 times dilution, 8 concentration points, DMSO content 1%. After the program is over, add 5 μL of the 1×9# dilution in the kit to each well, then add 5 μL of GST-labeled human Cereblon WT ligand, mix well, add 10 μL of HTRF detection reagent, and incubate at room temperature for 3 hours. The HTRF signal in each well was measured using an Envision plate reader. 100% binding inhibition was defined as the signal ratio under 200 μM standard lenalidomide treatment.

3. Data analysis

Calculate the ratio of acceptor and donor emission signals for each well:

Ratio＝Signal 665nm/Signal 620nm (ratio＝665nm signal/620nm signal)

CV(%)=Standard deviation/Mean Ratio (Deviation Coefficient (%)=Standard Deviation/Mean Ratio)

Cereblon binding inhibition%＝100%-100%*(Sample-Average_L)/(Average_H-L), that is, Cereblon binding inhibition rate (%)＝100%-100%×(Sample-L)/(H-L)

in:

H=Ave (DMSO treatment group);

L=Ave (200 μM lenalidomide standard treatment group).

The data was analyzed and processed by GraphPad Prism 9, the concentration-effect curve was fitted with a nonlinear four-parameter curve, and the IC ₅₀ of the compound was calculated:

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)*HillSlope))

in:

X: Log of cpd concentration (Log compound concentration);

Y:Percent inhibition(%inhibition)(inhibition percentage(%));

Bottom is the minimum suppression percentage;

Top is the maximum inhibition percentage;

HillSlope is the slope coefficient of the curve.

The binding ability of the disclosed compounds to Cereblon was determined by the above tests, and the measured IC ₅₀ values are shown in Table 1.

Table 1

化合物编号Compound number	IC ₅₀(μM) _IC50 (μM)
化合物1Compound 1	BB

A:<0.5μM; B:0.5-10μM; C:10-100μM; D:>100μM.

Test Example 2: MV-4-11 cell anti-proliferation activity experiment

1. Experimental instruments and materials

Instruments and Equipment

仪器名称equipment name	型号model	设备厂家Equipment manufacturers
离心机centrifuge	58005800	EppendorfEppendorf
细胞计数仪cell counter	IC1000IC1000	Count starCount star
倒置显微镜Inverted microscope	TS2TS2	NikonNikon
细胞培养箱cell culture incubator	1IVIOS 250i1IVIOS 250i	ThermoThermo
Envision酶标仪Envision microplate reader	Envision2105Envision2105	Perkin ElmerPerkin Elmer

Experimental reagents and consumables

2. Experimental steps

(1) Cell plating

The culture medium of the target cell MV-4-11 (CBP60522, Kebai) was removed, washed once with PBS, and then digested with trypsin-EDTA (0.25%) for 5 min. After digestion, add 10 mL of complete medium (IMDM+10% FBS) to neutralize the trypsin, pipette the cells, collect the cells, centrifuge at 1000 rpm for 5 min, count the cells, and adjust the cell density to 30,000 cells/mL. Take 90 μL of cell suspension and add it to a 96-well low adsorption plate. Add 200 μL of PBS to the edge wells of the 96-well plate. Centrifuge at 1000rpm for 5min to aggregate the cells into spheres and place them in a cell culture incubator for overnight culture.

(2) Cell dosing

The disclosed compound was dissolved in DMSO, and the stock concentration of the stock solution was 10 mM. Before administration, the compound was serially diluted 5-fold with DMSO, and a total of 8 gradient working solutions were used. Take 2 μL of each working solution of different concentrations, add it to the dilution plate of 198 μL medium, and mix well by pipetting. Take 10 μL of the medium containing the compound from the dilution plate and add it to the cell plate containing 90 μL of the cell suspension laid out the day before. nM. The positive control compound is dBET6. Add the diluted compound, 10 μL per well, and incubate at 37°C, 5% CO ₂ for 3 days.

(3)

Cell Viability Assay detection

The cells were taken out of the incubator and returned to room temperature for 30 min. Add 50 μL of

For the Cell Viability Assay reagent, shake and mix for 10 minutes and then read the plate with an Envision microplate reader.

3. Data analysis

The anti-proliferation activity of the disclosed compound on MV-4-11 cells is determined by the above test, and the cell growth inhibition rate of each sample well is calculated according to the original data.

Inhibition rate % = 100% * (1 - sample reading / reference average reading)

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)*HillSlope))

in:

X: Log compound concentration;

Y: inhibition rate (%);

Bottom is the minimum suppression percentage;

Top is the maximum inhibition percentage;

HillSlope is the slope coefficient of the curve.

Table 2 shows the antiproliferative activity of the disclosed compounds on MV-4-11 cells.

Table 2 Compounds of the present disclosure have antiproliferative activity on MV-4-11 cells

化合物编号Compound number	IC ₅₀(nM) IC ₅₀ (nM)
化合物2Compound 2	13.3613.36
化合物4Compound 4	15.9815.98

Test example 3: Experiment on the degradation activity of BRD4 protein in MV-4-11 cells

1. Experimental instruments and materials

Instruments and Equipment

仪器名称equipment name	设备厂家Equipment manufacturers	型号model
离心机centrifuge	EppendorfEppendorf	58005800
细胞计数仪cell counter	Count starCount star	IC1000IC1000
倒置显微镜Inverted microscope	NikonNikon	TS2TS2
细胞培养箱cell culture incubator	ThermoThermo	1IVIOS 250i1IVIOS 250i
电泳仪Electrophoresis	Bio-RadBio-Rad	16450501645050
转膜仪Film transfer instrument	Bio-RadBio-Rad	Trans-Blot Turbo(1704150)Trans-Blot Turbo(1704150)
Azure WB成像系统Azure WB imaging system	Azure biosystemsAzure biosystems	SapphireSapphire

Experimental reagents and consumables

2. Experimental steps

(1) Cell plating

The culture medium of MV-4-11 (CBP60522, Kebai) was removed, rinsed with PBS, and then digested with trypsin-EDTA (0.25%) for 5 min. After the digestion is complete, add 10 mL of complete medium (IMDM+10% FBS) to neutralize the trypsin, pipette the cells, collect the cells, centrifuge at 1000 rpm for 5 min, count the cells, and adjust the cell density to 500,000 cells/mL. Take 2mL of cell suspension and add it to a 12-well plate, and put it in a cell culture incubator for overnight culture.

(2) Cell dosing

The disclosed compound was dissolved in DMSO, and the stock concentration of the stock solution was 10 mM. The compound mother solution was serially diluted (4-fold dilution), with 8 concentration points, DMSO was used as a negative control group, and the DMSO content was 0.1%. Mix the compound and cell suspension and incubate at 37°C, 5% CO ₂ for 6 hours.

(3) Western Blot

After 6 hours of drug treatment, discard the supernatant, wash the cells with 1000 μL pre-cooled PBS, and then add 100 μL RIPA lysate (25 mM Tris-HCl, pH=7.5, 150 mM NaCl, 1% NP-40, 1 mM EDTA, pH=8.0 , 1mM PMSF, 1mM Na ₃ VO ₄ and 1x Protease Inhibitor Cocktail-P2714, Sigma), placed on ice for 10min to lyse. The samples after cell lysis were collected and centrifuged at 12,000 rpm at 4°C for 30 min, and the concentration of each protein sample was determined using a BCA kit for protein quantification. Add an appropriate amount of 5x SDS-PAGE protein loading buffer to the collected protein samples, and heat in a metal bath at 100°C for 10 minutes to fully denature the proteins. Load the protein sample into the sample hole of the SDS-PAGE gel, and electrophoresis at a constant voltage of 120V for 60min. After the electrophoresis, the gel was transferred to the PVDF membrane using a Bio-bad membrane transfer instrument. After the membrane was transferred, the bands were cut according to the size of the target protein, placed in 5% BSA for blocking at room temperature for 1 hour, and then BRD4 (1:3000 ) and β-actin (1:3000) antibodies, incubated overnight at 4°C. The antibody was recovered the next day, and the strip was washed 3 times with PBST (PBS containing 0.1% Tween-20), 10 min each time. After washing, the diluted Goat Anti-Rabbit IgG H&L (goat anti-rabbit secondary antibody) (1:3000) was added and incubated at room temperature for 1 h. The strips were washed 3 times with PBST (PBS containing 0.1% Tween-20). Use SuperSignal West Atto ultra-sensitive ECL luminescent fluid to detect protein.

3. Data analysis

The compound of the present disclosure has the BRD4 protein degradation activity in MV-4-11 cells determined by the above test, using the Image J software to read the gray value of the target sample band, and then calculate the protein degradation rate of each sample well according to the original data.

Degradation rate (%)=100%*(1-sample reading/DMSO reference reading)

in:

All raw data were analyzed and processed by GraphPad Prism 9, and the concentration-effect curve was fitted with a nonlinear four-parameter curve, and the DC ₅₀ of the compound was calculated:

Y＝Bottom+(Top-Bottom)/(1+10^((LogDC ₅₀ -X)*HillSlope))

in:

X: Log compound concentration;

Y: degradation rate (%);

Bottom is the minimum degradation percentage;

Top is the maximum degradation percentage;

HillSlope is the slope coefficient of the curve.

Table 3 shows the measured DC ₅₀ values of the disclosed compounds.

Table 3 Compounds of the present disclosure have activity on BRD4 protein degradation

化合物编号Compound number	最大降解率(％)Maximum degradation rate (%)	DC ₅₀(nM) DC ₅₀ (nM)
化合物2Compound 2	88.1388.13	3.373.37
化合物4Compound 4	78.5778.57	6.656.65

Test Example 4: Anti-proliferation activity experiment of CAL51 cells

1. Experimental instruments and materials

Instruments and Equipment

Experimental reagents and consumables

2. Experimental steps

(1) Cell plating

The culture medium of the target cell CAL51 (CBP60360, Kebai) was removed, washed once with PBS, and then digested with trypsin-EDTA (0.25%) for 5 min. After the digestion is complete, add 10 mL of complete medium (DMEM+10% FBS) to neutralize the trypsin, pipette the cells, collect the cells, centrifuge at 300 g for 5 min, count, and adjust the cell density to 40,000 cells/mL. Take 90uL of cell suspension and add it to a 96-well low adsorption plate. Add 200uL of PBS to the edge well. Centrifuge at 300g for 5min to aggregate the cells into spheres and put them in the cell culture incubator overnight.

(2) Cell dosing

The disclosed compound was dissolved in DMSO, and the stock concentration of the stock solution was 10 mM. Before administration, the DMSO gradient was diluted 10 times, and a total of 6 gradient working solutions were used. Take 2uL of each working solution of different concentrations, add it to the dilution plate of 198uL medium, and mix by pipetting. Take 10uL of the medium containing the compound from the dilution plate and add it to the cell plate containing 90uL of the cell suspension laid out the day before, and the final concentration of each gradient compound is 10,000, 1000, 100, 10, 1, 0.1nM. The positive control was CC-885. Add the diluted compound, 10uL per well, centrifuge and put it in a carbon dioxide incubator for 3 days.

(3)

3D Cell Viability Assay detection

The cells were taken out of the incubator and returned to room temperature for 30 min. Add 50uL of

3D Cell Viability Assay reagent, shake and mix for 10 minutes and then read the plate with Envision microplate reader.

3. Data analysis

The anti-proliferation activity of the disclosed compound on CAL51 cells is determined by the above test, and the cell growth inhibition rate of each sample well is calculated according to the original data.

Inhibition rate % = 100% * (1 - sample reading / reference average reading)

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)*HillSlope))

in:

X: Log compound concentration;

Y: inhibition rate (%);

Bottom is the minimum suppression percentage;

Top is the maximum inhibition percentage;

HillSlope is the slope coefficient of the curve.

Compound CC-885 is prepared with reference to 5.76 synthetic method in WO2008027542, and its structure is as follows:

Table 4 shows the antiproliferative activity of the disclosed compounds on CAL51 cells.

Table 4 Compounds of the present disclosure have antiproliferative activity on CAL51 cells

化合物编号Compound number	IC ₅₀(nM) IC ₅₀ (nM)
化合物3Compound 3	889889

Claims

A compound shown in formula (I) and pharmaceutically acceptable salts thereof:

in,

Y 1 , Y 2 , Y 3 are independently selected from CR 2 or N;

X 1 is selected from C, CR 1a or N;

X 2 is selected from CR 2a , CR 2a R 2b , N or NR 2a ;

X 3 is selected from O, S, C(=O), C(=S), S(=O) 2 , S(O), CR 3a R 3b or NR 3a ;

X 4 is selected from O, S, C(=O), C(=S), S(=O) 2 , S(O), CR 4a R 4b or NR 4a ;

R 1a , R 2a , R 2b , R 3a , R 3b , R 4a , R 4b are independently selected from H, halogen, CN, OH, NH 2 , -C(O) Rf , -C(O)N( R f ) 2 , -C(O)OR f , -C(O)ON(R f ) 2 , -NHC(O)R f , -NHC(O)OR f , -NHC(O)N(R f ) 2 , -S(O) 2 (R f ), C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, C 6 -C 10 aryl, 4-10 membered heterocyclyl or 5-10 OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, C 6 -C 10 aryl, 4-10 membered heterocyclyl or 5-10 membered heteroaryl Aryl is optionally substituted by R;

Or R 1a , R 4a and the cyclic group they are connected to form a carbon bridged ring or a bridged heterocyclic ring;

Or R 2a , R 3a and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

Or R 3a , R 4a and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

Or R 2a , R 2b and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

Or R 3a , R 3b and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

Or R 4a , R 4b and the atoms they are connected to together form a cycloalkyl or heterocyclic group;

R 1 is independently selected from halogen, CN, OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, 4-8 membered heterocyclyl, C 6 -C 10 aryl or 5- 10-membered heteroaryl, the OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, 4-8 membered heterocyclic group, C 6 -C 10 aryl or 5-10 membered Heteroaryl is optionally substituted by R;

R 2 is selected from H, halogen, CN, OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, 4-8 membered heterocyclyl, C 6 -C 10 aryl or 5- 10-membered heteroaryl, the OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, 4-8 membered heterocyclic group, C 6 -C 10 aryl or 5-10 membered Heteroaryl is optionally substituted by R;

Each R d is independently selected from halogen, CN, OH, NH 2 , SH, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-8 membered heterocyclyl;

Each R f is independently selected from H, halogen, CN, OH, NH 2 , C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl or 4-8 membered heterocyclyl;

n is independently selected from 0, 1 or 2.
The compound represented by formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein: X 2 is selected from CR 2a , CR 2a R 2b or NR 2a .
According to claim 1 or 2, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein: X 3 is selected from O, C(=O), CR 3a R 3b or NR 3a ; or, wherein X 3 is selected from C(=O) or CR 3a R 3b .
The compound represented by formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein: X 4 is selected from O, C(=O), CR 4a R 4b or NR 4a .
According to any one of claims 1-4, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein: R 1a , R 4a and the cyclic group connected to them form a carbon-bridged ring.
According to any one of claims 1-5, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein: R 2a , R 3a and the atoms they are connected to together form a C 3 -C 6 cycloalkyl group.
According to any one of claims 1-6, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein: R 3a , R 3b and the atoms they are connected to together form a C 3 -C 6 cycloalkyl group.
According to any one of claims 1-7, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein: R 1 is independently selected from halogen, CN, OH, NH 2 or C 1 -C 6 Alkyl, the OH, NH 2 or C 1 -C 6 alkyl is optionally substituted by R d .
According to any one of claims 1-8, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein: R 2 is selected from H, halogen, CN, OH, NH 2 or C 1 -C 6 Alkyl, the OH, NH 2 or C 1 -C 6 alkyl is optionally substituted by R d .
The compound represented by formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1-9, wherein: n is independently selected from 0 or 1.
According to the compound shown in formula (I) or its pharmaceutically acceptable salt according to any one of claims 1-10, wherein: the compound shown in formula (I) or its pharmaceutically acceptable salt is selected from formula (I) ') compound or a pharmaceutically acceptable salt thereof,

Wherein, X 1 is selected from CR 1a or N; X 2 is selected from CR 2a R 2b or NR 2a ;

R 1a , R 2a , R 2b , X 3 , X 4 , Y 1 , Y 2 , Y 3 , R 1 , and n are as defined in any one of claims 1-10.
According to claim 1, the compound shown in formula (I) or its pharmaceutically acceptable salt, wherein: the compound shown in formula (I) or its pharmaceutically acceptable salt is selected from the following compounds or its pharmaceutically acceptable salt Salt:
A compound represented by formula (II) and a pharmaceutically acceptable salt thereof:

Among them, [] indicates that L can be connected with X 2 , X 3 , X 4 , Y 1 , Y 2 or Y 3 ;

X 1 , X 2 , X 3 , X 4 , Y 1 , Y 2 , Y 3 , R 1 , n are as defined in claim 1;

When L is connected with Y 1 , Y 2 or Y 3 , Y 1 , Y 2 or Y 3 is C, when L is connected with X 2 , X 2 is C, CR 2a or N, when L is connected with X 3 , X 3 is CR 3a or N, when L is connected with X 4 , X 3 is CR 4a or N;

L is selected from the following structural units:

-(A L ) k -;

AL is independently selected from bond, -O-, -S-, -SO-, -SO 2 -, -SO 2 NR 10 -, -C(R 10 ) 2 -, -(CH 2 ) q -, - (O-CH 2 -CH 2 ) q -, -NR 10 -, -NR 10 C(=O)-, -C(=O)-, -C(=S)-, -OC(=O)- , -OC(O)NR 10 -, -C(O)ONR 10 -, -CR 10 = CR 10 -, -C≡C-, -P(O)R 10 -, -P(O)OR 10 - ,
C 3 -C 10 cycloalkyl, 4-10 membered heterocyclic group, C 6 -C 10 aryl or 5-10 membered heteroaryl, said C 3 -C 10 cycloalkyl, 4-10 membered heterocyclic Base, C 6 -C 10 aryl or 5-10 membered heteroaryl are optionally substituted by R 11 ;

R 10 is selected from H, OH, NH 2 or C 1 -C 6 alkyl;

R 11 is selected from halogen, CN, OH, NH 2 or C 1 -C 6 alkyl;

k is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

q is selected from 0, 1, 2, 3, 4, 5 or 6.
According to claim 13, the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, wherein: AL is independently selected from bond, -O-, -S-, -C(R 10 ) 2 -, - (CH 2 ) q -, -(O-CH 2 -CH 2 ) q -, -NR 10 -, -NR 10 C(=O)-, -C(=O)-, C 3 -C 10 cycloalkane base, 4-10 membered heterocyclic group, C 6 -C 10 aryl group or 5-10 membered heteroaryl group, the C 3 -C 10 cycloalkyl group, 4-10 membered heterocyclic group, C 6 -C 10 Aryl or 5-10 membered heteroaryl is optionally substituted by R 11 ;

Alternatively, AL is independently selected from a bond, -O-, -C(R 10 ) 2 -, -(CH 2 ) q -, -(O-CH 2 -CH 2 ) q -, -NR 10 -, - NR 10 C(=O)-, -C(=O)-, 5-6 membered heterocyclic group or 5-6 membered heteroaryl group, the 5-6 membered heterocyclic group or 5-6 membered heteroaryl group optionally substituted by R 11 ;

Alternatively, AL is independently selected from a bond, -O-, -C(R 10 ) 2 -, -(CH 2 ) q -, -(O-CH 2 -CH 2 ) q -, -NR 10 -, - NR 10 C(=O)-, -C(=O)-, piperazinyl or triazolyl;

Alternatively, AL is independently selected from -O-, -(CH 2 ) q -, -(O-CH 2 -CH 2 ) q -, -NR 10 -, -NR 10 C(=O)-, -C (=O)-, C 3 -C 10 cycloalkyl, 5-6 membered heterocyclic group, 5-6 membered heteroaryl or C 6 -C 10 aryl, said C 3 -C 10 cycloalkyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl or C 6 -C 10 aryl is optionally substituted by R 11 .
According to the compound represented by formula (II) or a pharmaceutically acceptable salt thereof according to claim 13 or 14, wherein: L is selected from the following structural units:

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-(CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(CH 2 ) q -NHC(=O)-(CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(5-10 heteroaryl)-(CH2) q -C(=O)NH-(CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-(CH 2 ) q -NR 10 -;

-(O-CH 2 -CH 2 ) q -(5-10 heteroaryl)-(CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-;

-NR 10 -(CH 2 ) q -(5-10heteroaryl)-(CH 2 ) q -O-(CH 2 ) q -C(=O)-(CH 2 ) q -NR 10 -;

-NR 10 -(CH 2 ) q -(CH 2 ) q -NHC(=O)-;

-NR 10 -(CH 2 ) q -(5-10 heteroaryl)-(CH 2 ) q -NR 10 -;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -NR 10 -;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -O-(CH 2 ) q -C(=O)-;

-NR 10 -(CH 2 ) q -(5-10heteroaryl)-(CH 2 ) q -O-(CH 2 ) q -C(=O)-(CH 2 ) q -;

-(4-10 membered heterocyclyl)-C(=O)-(CH 2 ) q -C(=O)NH-(CH 2 ) q -NR 10 -;

-(5-10 Heteroaryl)-C(=O)-(CH 2 ) q -C(=O)NH-(CH 2 ) q -NR 10 -;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -;

-O-(CH 2 ) q -(5-10 heteroaryl)-C(=O)-;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-(CH 2 ) q -NR 10 -;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-;

-O-(CH 2 ) q -(CH 2 ) q -;

-NR 10 -(CH 2 ) q -(CH 2 ) q -;

-NR 10 -(CH 2 ) q -(CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(CH 2 ) q -NR 10 -;

-O-(CH 2 ) q -(CH 2 ) q -O-;

-O-(CH 2 ) q -(O-CH 2 -CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-(CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(O-CH 2 -CH 2 ) q -(CH 2 ) q -C(=O)-(CH 2 ) q -NHC(=O)-(CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(4-10 membered heterocyclyl)-(CH 2 ) q -NR 10 -;

-NR 10 -(CH 2 ) q -(4-10 membered heterocyclyl)-(CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(4-10 membered heterocyclyl)-(CH 2 ) q -O-;

-NR 10 -(CH 2 ) q -(4-10 membered heterocyclyl)-(CH 2 ) q -(CH 2 ) q -O-;

-OC(=O)-(C 3 -C 10 cycloalkyl)-(CH 2 ) q -O-;

-OC(=O)-(C 3 -C 10 cycloalkyl)-(CH 2 ) q -NH-;

-OC(=O)-(C 3 -C 10 cycloalkyl)-(CH 2 ) q -;

-(CH 2 ) q -NHC(=O)-NH-(C 6 -C 10 aryl)-;

-NR 10 -(CH 2 ) q -(CH 2 ) q -C(=O)-;

-NR 10 -(CH 2 ) q -C(=O)NH-(CH 2 ) q -;

The C 3 -C 10 cycloalkyl, C 6 -C 10 aryl, 5-10 heteroaryl or 4-10 membered heterocyclic group is optionally substituted by R 11 ,

q is as defined in claim 13 or 14.
According to the compound represented by formula (II) or a pharmaceutically acceptable salt thereof according to any one of claims 13-15, wherein: L is selected from the following structural units:

-NH-(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-(CH 2 ) q -O-;

-NH-(CH 2 ) q -(CH 2 ) q -NHC(=O)-(CH 2 ) q -O-;

-NH-(CH 2 ) q -triazolyl-(CH 2 ) q -C(=O)NH-(CH 2 ) q -O-;

-NH-(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-(CH 2 ) q -NH-;

-(O-CH 2 -CH 2 ) q -triazolyl-(CH 2 ) q -O-;

-O-(CH 2 ) q -(CH 2 ) q -O-;

-NH-(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-;

-NH-(CH 2 ) q -triazolyl-(CH 2 ) q -O-(CH 2 ) q -C(=O)-(CH 2 ) q -NH-;

-NH-(CH 2 ) q -(CH 2 ) q -NHC(=O)-;

-O-(CH 2 ) q -(CH 2 ) q -;

-NH-(CH 2 ) q -triazolyl-(CH 2 ) q -NH-;

-NH-(CH 2 ) q -(O-CH 2 -CH 2 ) q -NH-;

-NH-(CH 2 ) q -(O-CH 2 -CH 2 ) q -O-(CH 2 ) q -C(=O)-;

-NH-(CH 2 ) q -triazolyl-(CH 2 ) q -O-(CH 2 ) q -C(=O)-(CH 2 ) q -;

-piperazinyl-C(=O)-( CH2 ) q -C(=O)NH-( CH2 ) q -NH-;

-NH-(CH 2 ) q -(O-CH 2 -CH 2 ) q -O-;

-NH-(CH 2 ) q -(O-CH 2 -CH 2 ) q -;

-O-(CH 2 ) q -triazolyl-C(=O)-;

-NH-(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-(CH 2 ) q -NH-;

-NH-(CH 2 ) q -(O-CH 2 -CH 2 ) q -NHC(=O)-;

-OC(=O)-cyclohexyl-( CH2 ) q -O-;

-OC(=O)-cyclohexyl-( CH2 ) q -NH-;

-OC(=O)-cyclohexyl-(CH 2 ) q -;

-Phenyl-NHC(=O)-NH-(CH 2 ) q -;

-NH-(CH 2 ) q -(CH 2 ) q -C(=O)-;

-NH-(CH 2 ) q -C(=O)NH-(CH 2 ) q -;

The triazolyl, piperazinyl, cyclohexyl or phenyl is optionally substituted by R,

q is as defined in any one of claims 13-15.
The compound represented by formula (II) or a pharmaceutically acceptable salt thereof according to any one of claims 13-16, wherein: k is selected from 1, 2, 3, 4, 5, 6, 7, 8 or 9 .
According to any one of claims 13-17, the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, wherein: the compound represented by formula (II) or a pharmaceutically acceptable salt thereof is selected from the group consisting of formula (II) ') compound or a pharmaceutically acceptable salt thereof,

Wherein, X 1 is selected from CR 1a or N; X 2 is selected from CR 2a R 2b or NR 2a ;

R 1a , R 2a , R 2b , X 3 , X 4 , Y 1 , Y 2 , Y 3 , R 1 , n, L, [] are as defined in any one of claims 13-17.
According to claim 13, the compound shown in formula (II) or its pharmaceutically acceptable salt, wherein: the compound shown in formula (II) or its pharmaceutically acceptable salt is selected from the following compounds or its pharmaceutically acceptable salt Salt:
A pharmaceutical composition, which comprises the compound represented by formula (I) or formula (II) or a pharmaceutically acceptable salt thereof according to any one of claims 1-19, and pharmaceutically acceptable auxiliary materials.
The compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof in any one of claims 1-19, or the pharmaceutical composition of claim 20 is used in the preparation of prevention or treatment of related diseases mediated by CRBN use in medicines.
The compound shown in any one of claims 1-19 formula (I) or (II) or a pharmaceutically acceptable salt thereof, as the synthetic intermediate of the bifunctional compound targeting protein degradation, in the preparation of targeting protein degradation Use of bifunctional compounds.
A compound represented by formula (III) and pharmaceutically acceptable salts thereof:

Wherein, [], X 1 , X 2 , X 3 , X 4 , Y 1 , Y 2 , Y 3 , R 1 , L, n are as defined in any one of claims 13-17;

The PTM is a targeting protein binding moiety.
According to claim 23, the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, wherein: PTM is selected from the following binding parts of targeting proteins: ALK, AR, BET1, BRAF, BRCA2, BRD4, BRD9, BTK , CBL, CCNE1, CCNE2, CCR4, CCR7, CCR9, CD47, CLDN18, CYP, DDR1, DMPK, EGFR, ERBB2, ERBB3, ERBB4, FGFR1, FGFR2, FGFR3, FGFR4, GSPT1, JAK1, JAK3, KIF18A, KRAS, LCK , MET, NTRK1, NTRK2, NTRK3, PCSK9, PKMYT1, PARP7, PARP14, RAD51, RBM10, RET, RORA, STAT3, SOS1, TYK2, USP1 or USP14.
According to claim 23, the compound represented by formula (III) or its pharmaceutically acceptable salt, wherein: the compound represented by formula (III) or its pharmaceutically acceptable salt is selected from the following compounds or its pharmaceutically acceptable salt Salt:
A pharmaceutical composition, comprising the compound represented by formula (III) or a pharmaceutically acceptable salt thereof according to any one of claims 23-25, and pharmaceutically acceptable auxiliary materials.
The compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof according to any one of claims 1-19, or the pharmaceutical composition described in claim 20, or the pharmaceutical composition described in any one of claims 23-25 Use of the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 26 in the preparation of medicines for preventing or treating abnormal cell proliferation diseases.