Compounds as VAV1 degradation agents and use thereof

Abstract

Provided in the present invention are compounds as VAV1 degradation agents and the use thereof. Specifically provided are compounds represented by formula (II), and tautomers, stereoisomers, pharmaceutically acceptable salts or prodrugs thereof. The compounds provided by the present invention have good efficacy, and can be used for preparing drugs for treating or preventing VAV1-related diseases.

Description

Compounds as vav1 degraders and uses thereof

[0001] Priority information

[0002] The present disclosure claims priority to and the benefit of Chinese Patent Application No. 202411162599.9, filed August 22, 2024, Chinese Patent Application No. 202411547472.9, filed October 31, 2024, and Chinese Patent Application No. 2025110558012, filed July 29, 2025, to the Chinese National Intellectual Property Office, and incorporates by reference the entire contents of each of the foregoing applications herein. TECHNICAL FIELD

[0003] The present application provides a class of compounds as VAV1 degraders and uses thereof. Specifically provided are compounds represented by formula (II), tautomers, stereoisomers, pharmaceutically acceptable salts or prodrugs thereof. The compounds provided by the present application have good drug efficacy and can be prepared for drugs for treating or preventing VAV1 related diseases. BACKGROUND

[0004] Drug-induced targeted protein degradation (TPD) technology is an emerging drug treatment strategy. This technology takes advantage of two major protein degradation systems (ubiquitin proteasome system (UPS) and lysosomal degradation system) naturally present in the human body to destroy and degrade key proteins related to diseases, thereby achieving the effect of treating diseases. Currently, molecular glue, proteolytic targeting chimera, lysosome targeting chimera, autophagy targeting chimera, autophagy linking compound, and other technologies have been developed. Among them, multiple drugs based on the principle of molecular glue have been approved for marketing, and have achieved good efficacy. The success of molecular glue in clinical practice and its future potential have prompted pharmaceutical companies to pay increasing attention to the development of related drugs.

[0005] VAV family proteins, including VAV1, VAV2, and VAV3, are Rho family GTPase guanine nucleotide exchange factors (GEFs). VAV1 is a 95-kDa protein that is a positive regulator of T cell receptor (TCR) and B cell receptor (BCR) signaling. VAV1 is mainly expressed in hematopoietic cells of the human body, including T cells, B cells, monocytes, natural killer (NK) cells, granulocytes, and dendritic cells, while its family members VAV2 and VAV3 are more widely expressed. VAV1 is rapidly phosphorylated upon a variety of stimuli, such as stimulation of T cell receptors, B cell receptors, and various cytokine receptors. In hematopoietic-derived cells, such as T cells, B cells, natural killer cells, and osteoclasts, VAV1 regulates a variety of cellular functions and signaling pathways by activating certain GTPases. VAV1-mediated functions include gene transcription, development and activation of immune cells such as T cells and B cells.

[0006] Genome-wide CRISPR-Cas9 screening confirmed that VAV1 is an important positive regulator of T cell activation / function, and VAV1 promotes the proliferation of cells sensitive to TCR signaling pathway, such as human Jurkat T cells and primary human CD4+ and CD8+ T cells. In addition, VAV1 knockout mouse data show that VAV1 plays a key role in T / B lymphocyte function and antigen receptor signaling, especially in T cells. More importantly, VAV1 -deficient mouse thymocytes and splenic T cells show multiple defects in TCR-mediated signaling, such as impaired calcium (Ca 2+ ) mobilization and transcription factor activation. However, T cells with VAV1 GEF activity loss (VAV1 L334A / K335A) exhibit normal TCR-mediated Ca 2+ flux and nuclear factor of activated T cells (NFAT) activation. It is shown that both GEF activity and scaffold function of VAV1 play an important role in the TCR signaling pathway.

[0007] Genetic analysis found that rodents carrying VAV1 R63W variation showed lower susceptibility in experimental autoimmune encephalomyelitis (EAE) and palmitoyl-induced arthritis compared with wild type (WT). In the mouse model of antigen (methylated bovine serum albumin)-induced arthritis (AIA), VAV1 knockout mice showed less disease symptoms (such as inflammation, synovial thickening and cartilage degradation), reduced T cell proliferation and reduced joint infiltration of CD4+ T cells, neutrophils and macrophages than wild type mice. This again indicates that VAV1 plays an important role in T cell differentiation and function. Therefore, VAV1 can be a therapeutic target for immune-mediated diseases. SUMMARY

[0008] In a first aspect of the present application, the present application provides a compound represented by formula (II), a tautomer, a stereoisomer, a pharmaceutically acceptable salt or a prodrug thereof,

[0009] wherein,

[0010] R1 is halogen, OCF3, CN, C 2-6 alkyl, -OC 2-6 alkyl, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, 4-10 membered heterocycloalkyl, said C 2-6 alkyl, -OC 2-6 alkyl, C 3-6 cycloalkyl, C 2-6 alkynyl and 4-10 membered heterocycloalkyl are each independently optionally substituted with 1, 2, 3 or 4 R 1a ;

[0011] each R 1a is independently H, halogen, OH, NH2, CN, C 1-6 alkyl or haloC 1-6 alkyl;

[0012] R2, R3and R4are each independently H, halogen, NH2, CN, C 1-6 alkyl or haloC 1-6 alkyl;

[0013] X1is N or CR5;

[0014] X2is N or CR 51 ;

[0015] X3is N or CR7;

[0016] X4is N or CR 71 ;

[0017] R5, R 51 , R7and R 71 are each independently H, halogen, NH2, CN or C 1-6 alkyl or haloC 1-6 alkyl;

[0018] R8is H, D, halogen or C 1-6 alkyl;

[0019] L1is a single bond, -O-, -NH-, -O-C 1-3 alkyl-, -NH-C 1-3 alkyl-, -C 1-3 alkyl- or -C 3-6 cycloalkyl-, wherein said -NH-, -O-C 1- 3alkyl-, -NH-C 1-3 alkyl-, -C 1-3 alkyl- and -C 3-6 cycloalkyl- is independently optionally substituted with 1, 2, 3 or 4 R 1L ;

[0020] each R 1L is independently H, halogen, C 1-3 alkyl or C 3-6 cycloalkyl;

[0021] R6is CN, -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, -C 0-3 alkyl-S(=O)(=NR62 )R 63 C 1-6 Alkyl, 4-10-membered heterocyclic alkyl, 5-10-membered heterocyclic alkenyl, 6-10-membered aryl or 5-10-membered heteroaryl, wherein C 1-6 Alkyl, 4-10-membered heterocyclic alkyl, 5-10-membered heterocyclic alkenyl, 6-10-membered aryl, and 5-10-membered heteroaryl groups are each independently and optionally bounded by 1, 2, 3, 4, 5, 6, 7, or 8 R groups. 6a replace;

[0022] R 61 R 62 and R 63 H and C are independent of each other. 1-6 Alkyl or C 3-6 cycloalkyl;

[0023] Each R 6a The independent components are H, halogen, oxo (=O), thio (=S), OH, NH2, CN, and C, respectively. 1-6 Alkyl, Halogenated C 1-6 Alkyl or C 3-6 cycloalkyl;

[0024] Or, two Rs 61 Together with the N, P, or S atoms to which they are attached, they form a 4-10 membered heterocyclic alkyl group, wherein the 4-10 membered heterocyclic alkyl group is optionally surrounded by 1, 2, 3, or 4 R atoms. 6b replace;

[0025] Or, R 62 With R 63 Together with the N and S atoms attached thereto, they form a 5-10 membered heterocyclic alkenyl group, which is optionally surrounded by 1, 2, 3 or 4 R atoms. 6b replace;

[0026] Each R 6b They are, independently, H, halogen, oxo (=O), OH, NH2, CN, and C, respectively. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-6 cycloalkyl, -L2-C 1-6 Alkyl, -L2-C 3-6 Cycloalkyl or -L2-4-8-membered heterocycloalkyl, wherein C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-6 cycloalkyl, -L2-C 1-6 Alkyl, -L2-C 3-6 The cycloalkyl or -L2-4-8-membered heterocycloalkyl groups are each optionally substituted by 1, 2, 3 or 4 Rs, independently;

[0027] each L2is independently -0-, -N(R 2L )-, -S-, -S(=0)-, -S(=0)2-, or -C(=0)-;

[0028] R 2L is H or C 1-3 alkyl;

[0029] each R is independently H, halogen, OH, NH2, or CN;

[0030] when R1is halogen, R6is -C 0-3 alkyl P(=0)(R 61 )2, -C 0-3 alkyl-(N=)S(=0)(R 61 )2, -C 0-3 alkyl-S(=0)(=NR 62 )R 63 , 4-10 membered heterocycloalkyl, or 5-10 membered heterocycloalkenyl, each of which is independently optionally substituted with 1, 2, 3, or 4 R

[0031] The heteroatom groups in the "heterocycloalkyl", "heterocycloalkenyl", and "heteroaryl" groups comprise N, O, S, S(=0), S(=0)2, or S(=0)(=NH), the number of which is 1, 2, 3, or 4; when the number of the heteroatom groups is more than one, the heteroatom groups are the same or different.

[0032] In an optional embodiment of the present application, the above-mentioned compound, its tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug, the compound having the structure of formula (II):

[0033] wherein,

[0034] R1is halogen, OCF3, CN, C 2-6 alkyl, -OC 2-6 alkyl, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, 4-10 membered heterocycloalkyl, each of which is independently optionally substituted with 1, 2, 3, or 4 R 2-6 alkyl, -OC 2-6 alkyl, C 3-6 cycloalkyl, C 2-6 alkynyl, and 4-10 membered heterocycloalkyl are each independently optionally substituted with 1, 2, 3, or 4 R 1a ;

[0035] each R 1a is independently H, halogen, OH, NH2, CN, C 1-6alkyl or haloC 1-6 alkyl;

[0036] R2, R3and R4are each independently H, halogen, NH2, CN, C 1-6 alkyl or haloC 1-6 alkyl;

[0037] X1is N or CR5;

[0038] X2is N or CR 51 ;

[0039] X3is N or CR7;

[0040] X4is N or CR 71 ;

[0041] R5, R 51 , R7and R 71 are each independently H, halogen, NH2, CN or C 1-6 alkyl or haloC 1-6 alkyl;

[0042] R8is H, D, halogen or C 1-6 alkyl;

[0043] L1is a single bond, -O-, -NH-, -O-C 1-3 alkyl-, -NH-C 1-3 alkyl- or -C 1-3 alkyl-, wherein said -NH-, -O-C 1-3 alkyl-, -NH-C 1-3 alkyl- and -C 1-3 alkyl- are each independently optionally substituted with 1, 2, 3 or 4 R 1L ;

[0044] each R 1L is independently H, halogen, C 1-3 alkyl or C 3-6 cycloalkyl;

[0045] R6is CN, -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, -C 0-3 alkyl-S(=O)(=NR 62 )R 63 , C 1-6 alkyl, 4-10 membered heterocycloalkyl, 5-10 membered heterocycloalkenyl, 6-10 membered aryl or 5-10 membered heteroaryl, said C 1-6alkyl, 4-10 membered heterocycloalkyl, 5-10 membered heterocycloalkenyl, 6-10 membered aryl, and 5-10 membered heteroaryl are each independently optionally substituted with 1, 2, 3, or 4 R 6a substituted;

[0046] R 61 , R 62 , and R 63 are each independently H or C 1-6 alkyl;

[0047] each R 6a is independently H, halogen, oxo (=0), thioxo (C=S), OH, NH2, CN, C 1-6 alkyl, haloC 1-6 alkyl, or C 3- 6 cycloalkyl;

[0048] Alternatively, two R 61 and the P or S atom to which they are attached together form a 5-10 membered heterocycloalkyl group, which is optionally substituted with 1, 2, 3, or 4 R 6b substituted;

[0049] each R 6b is independently H, halogen, oxo (=0), OH, NH2, CN, C 1-6 alkyl, haloC 1-6 alkyl, C 3-6 cycloalkyl, -L2-C 1-6 alkyl, -L2-C 3-6 cycloalkyl, or -L2-4-8 membered heterocycloalkyl, each independently optionally substituted with 1, 2, 3, or 4 R 1-6 alkyl, haloC 1-6 alkyl, C 3-6 cycloalkyl, -L2-C 1-6 alkyl, -L2-C 3-6 cycloalkyl, or -L2-4-8 membered heterocycloalkyl, each independently optionally substituted with 1, 2, 3, or 4 R

[0050] each L2is independently -0-, -N(R 2L )-, -S-, -S(=0)-, -S(=0)2-, or -C(=0)-;

[0051] R 2L is H or C 1-3 alkyl;

[0052] each R is independently H, halogen, OH, NH2, or CN;

[0053] when R1is halogen, R6is -C 0-3 alkyl P(=0)(R 61 )2, -C0-3 alkyl-(N=)S(=0)(R 61 )2, -C 0-3 alkyl-S(=0)(=NR 62 )R 63 or 5-10 membered heterocycloalkenyl, which is substituted with at least one thioxo (C=S);

[0054] The heteroatom groups in the "heterocycloalkyl", "heterocycloalkenyl" and "heteroaryl" groups comprise N, O, S, S(=0), S(=0)2or S(=0)(=NH), the number of which is 1, 2, 3 or 4; when the number of the heteroatom groups is plural, the heteroatom groups are the same or different.

[0055] In an optional embodiment of the present application, the above-mentioned compound, its tautomer, stereoisomer, pharmaceutically acceptable salt or prodrug, the compound has the structure of formula (I)

[0056] wherein,

[0057] R1is halogen, OCF3, CN, C 2-6 alkyl, -OC 2-6 alkyl, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, 4-10 membered heterocycloalkyl, the C 2-6 alkyl, -OC 2-6 alkyl, C 3-6 cycloalkyl, C 2-6 alkynyl and 4-10 membered heterocycloalkyl are each independently optionally substituted with 1, 2, 3 or 4 R 1a ;

[0058] each R 1a is independently H, halogen, OH, NH2, CN, C 1-6 alkyl or halogenated C 1-6 alkyl;

[0059] R2, R3and R4are each independently H, halogen, NH2, CN, C 1-6 alkyl or halogenated C 1-6 alkyl;

[0060] R5and R7are each independently H, halogen, NH2, CN or C 1-6 alkyl or halogenated C 1-6 alkyl;

[0061] R8is H, D, halogen or C 1-6 alkyl;

[0062] L1is a single bond, -O-, -NH-, -O-C 1-3 alkyl-, -NH-C 1-3 alkyl- or -C 1-3 alkyl, wherein said -NH-, -O-C 1-3 alkyl-, -NH-C 1-3 alkyl- and -C 1-3 alkyl are each independently optionally substituted with 1, 2, 3 or 4 R 1L substituents;

[0063] each R 1L is independently H, halogen, C 1-3 alkyl or C 3-6 cycloalkyl;

[0064] R6is -P(=O)(R 61 )2, -(N=)S(=O)(R 61 )2, -S(=O)(=NR 62 )R 63 , CN, C 1-6 alkyl, 4-10 membered heterocycloalkyl, 5-10 membered heterocycloalkenyl, 6-10 membered aryl or 5-10 membered heteroaryl, said C 1-6 alkyl, 4-10 membered heterocycloalkyl, 5-10 membered heterocycloalkenyl, 6-10 membered aryl and 5-10 membered heteroaryl are each independently optionally substituted with 1, 2, 3 or 4 R 6a substituents;

[0065] R 61 , R 62 and R 63 are each independently H or C 1-6 alkyl;

[0066] each R 6a is independently H, halogen, oxo (=O), OH, NH2, CN, C 1-6 alkyl or halogenated C 1-6 alkyl;

[0067] or, two R 61 and the P or S atom to which they are attached together form a 5-10 membered heterocycloalkyl;

[0068] when R1is halogen, R6is -P(=O)(R 61 )2, -(N=)S(=O)(R 61 )2or -S(=O)(=NR 62 )R 63 ;

[0069] The heteroatom groups in the "heterocycloalkyl", "heterocycloalkenyl" and "heteroaryl" groups comprise N, O, S, S(=O), S(=O)2or S(=O)(=NH), the number of which is 1, 2, 3 or 4; when the number of the heteroatom groups is plural, the heteroatom groups are the same or different.

[0070] In an optional embodiment of the present application, the above R1is Cl, CN, -OCF3, -C≡C-CH3, 2-4 alkyl, C 3-4 alkyl, C 2-4 alkyl, C 3-4 alkyl, C 1a substituted.

[0071] In an optional embodiment of the present application, the above R1is Cl, CN, -OCF3, -C≡C-CH3,

[0072] In an optional embodiment of the present application, each of the above R 1a is independently H or F.

[0073] In an optional embodiment of the present application, the above R2, R3and R4are each independently H.

[0074] In an optional embodiment of the present application, the above R5, R 51 , R7and R 71 are each independently H.

[0075] In an optional embodiment of the present application, the above R5and R7are each independently H.

[0076] In an optional embodiment of the present application, the above R8is H or D.

[0077] In an optional embodiment of the present application, the above L1is a single bond, -C 1-3 alkyl-, -O-C 1-3 alkyl-, -O-C 3-6 alkyl-.

[0078] In an optional embodiment of the present application, the above L1is a single bond, -CH2-, -O-CH2- or

[0079] In an optional embodiment of the present application, the above L1is a single bond, -CH2- or -O-CH2-.

[0080] In an optional embodiment of the present application, each R6above is 4-8 membered heterocycloalkyl, 5-8 membered heterocycloalkenyl, or 5-6 membered heteroaryl, each of which is independently optionally substituted with 1, 2, 3, 4, or 5 R 6a substituents.

[0081] In an optional embodiment of the present application, each R 6a is independently H, oxo (=0), thioxo (=S), CH3, -CH2CH3, CHF2, CF3, or cyclopropyl.

[0082] In an optional embodiment of the present application, each R 6a is independently H, oxo (=0), thioxo (=S), CH3, CHF2, or cyclopropyl.

[0083] In an optional embodiment of the present application, each R 6a is independently H, oxo (=0), CH3, or CHF2.

[0084] In an optional embodiment of the present application, R6above is

[0085] In an optional embodiment of the present application, R6above is

[0086] In an optional embodiment of the present application, R6above is

[0087] In an optional embodiment of the present application, when R1is Cl, R6is -C 0-3 alkyl-P(=0)(R 61 )2, -C 0-3 alkyl-(N=)S(=0)(R 61 )2, or -C 0-3 alkyl-S(=0)(=NR 62 )R 63 .

[0088] In an optional embodiment of the present application, when R1is Cl, R6is 5-8 membered heterocycloalkenyl, which is substituted with at least one thioxo (=S). In an optional embodiment of the present application, the above when R1is Cl, R6is -P(=0)(R 61 )2, -(N=)S(=0)(R 61 )2, or -S(=0)(=NR 62 )R 63.

[0089] In an optional embodiment of the present application, the above R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, or -C 0-3 alkyl-S(=O)(=NR 62 )R 63 .

[0090] In an optional embodiment of the present application, the above R6is -P(=O)(R 61 )2, -(N=)S(=O)(R 61 )2, or -S(=O)(=NR 62 )R 63 .

[0091] In an optional embodiment of the present application, the above R 61 , R 62 , and R 63 are each independently H, CH3, -CH2CH3, or cyclopropyl.

[0092] In an optional embodiment of the present application, the above R 61 , R 62 , and R 63 are each independently H or CH3.

[0093] In an optional embodiment of the present application, the above R6is

[0094] In an optional embodiment of the present application, the above R6is

[0095] In an optional embodiment of the present application, the above R6is

[0096] In an optional embodiment of the present application, the above two R 61 , together with the P or S atom to which they are attached, form a 4-6 membered heterocycloalkyl, which is optionally substituted with 1, 2, 3, or 4 R 6b .

[0097] In an optional embodiment of the present application, the above two R 61 , together with the P or S atom to which they are attached, form a heterocycloalkyl, which is a 5-6 membered heterocycloalkyl.

[0098] In an optional embodiment of the present application, the above R 62With R 63 Together with the N and S atoms attached thereto, they form a 5-10 membered heterocyclic alkenyl group, which is optionally surrounded by 1, 2, 3 or 4 R atoms. 6b replace.

[0099] In an optional embodiment of the present invention, the above-mentioned R 6b They are, independently, H, halogen, oxo (=O), OH, NH2, CN, and C, respectively. 1-3 Alkyl, Halogenated C 1-6 Alkyl, C 3-6 cycloalkyl, -L2-C 1-3 Alkyl, -L2-C 3-5 Cycloalkyl or -L2-4-6-membered heterocycloalkyl, wherein C 1-3 Alkyl, Halogenated C 1-6 Alkyl, C 3-6 cycloalkyl, -L2-C 1-3 Alkyl, -L2-C 3-5 The cycloalkyl or -L2-4-6-membered heterocycloalkyl groups are each optionally substituted by 1, 2, 3 or 4 Rs, respectively.

[0100] In an optional embodiment of the present invention, the above-mentioned R 6b H and -C (=O)-C are independently represented respectively. 1-3 Alkyl, or -C(=O)-C 3-5 Cycloalkyl.

[0101] In an optional embodiment of the present invention, the above-mentioned R 6b They are independently H, -C(=O)-CH3, -C(=O)-CH2CH3 or -C(=O)-cyclopropyl.

[0102] In an optional embodiment of the present invention, each of the above-mentioned L2s is independently -O-, -N(R) 2L )- or -C(=O)-.

[0103] In an optional embodiment of the present invention, the above-mentioned R 2L For H.

[0104] In an optional embodiment of the present invention, R6 is -C. 0-3 Alkyl-P(=O)(R) 61 )2 or -C 0-3 Alkyl-(N=)S(=O)(R 61 )2, where two R 61 When R6 forms a heterocyclic alkyl group together with the P or S atom it is attached to, R6 is...

[0105] In an optional embodiment of the present invention, R6 is -C. 0-3 Alkyl-S(=O)(=NR) 62 )R 63 , where R 62 With R 63 When R6 forms a 5-10 membered heterocyclic alkenyl group together with the N and S atoms it is attached to, R6 is

[0106] In an optional embodiment of the present invention, R6 is -C. 0-3 Alkyl-P(=O)(R) 61 )2、-C 0-3 Alkyl-(N=)S(=O)(R 61 2. 4-10-membered heterocyclic alkyl or 5-10-membered heterocyclic alkenyl, wherein two R 61 When R6 forms a heterocyclic alkyl group together with the P or S atom it is attached to, R6 is...

[0107] In an optional embodiment of the present invention, R6 is -C. 0-3 Alkyl-P(=O)(R) 61 )2、-C 0-3 Alkyl-(N=)S(=O)(R 61 )2 or 5-10 membered heterocyclic alkenyl groups, wherein two R 61 When R6 forms a heterocyclic alkyl group together with the P or S atoms it is attached to, R6 is...

[0108] In an optional embodiment of the present invention, R6 is...

[0109] In an optional embodiment of the present invention, R6 is...

[0110] In an optional embodiment of the present invention, the above-mentioned two R 61 When R6 forms a heterocyclic alkyl group together with the P or S atoms it is attached to, R6 is...

[0111] In an optional embodiment of the present invention, R6 is...

[0112] In an optional embodiment of the present invention, R6 is...

[0113] In an alternative embodiment of the application, R6is as defined above, and R1is Cl.

[0114] In an alternative embodiment of the application, R6is a 5-8 membered heterocycloalkenyl group, said 5-8 membered heterocycloalkenyl group being optionally substituted with 1, 2, 3 or 4 R 6a , wherein at least one R 6a is thioxo (=S).

[0115] In an alternative embodiment of the application, when R1is Cl, R6is

[0116] In an alternative embodiment of the application, when R1is Cl, R6is

[0117] In an alternative embodiment of the application, the above compound is selected from the following structures:

[0118] said R1, R8being as defined in the application, wherein L1and R6satisfy one or more of the following conditions:

[0119] (1c) L1is a single bond, -CH2-, -O-CH2- or

[0120] (2c) R6is a 4-8 membered heterocycloalkyl group, a 5-8 membered heterocycloalkenyl group or a 5-6 membered heteroaryl group, said 4-8 membered heterocycloalkyl group, 5-8 membered heterocycloalkenyl group and 5-6 membered heteroaryl group being independently of each other optionally substituted with 1, 2, 3, 4 or 5 R 6a , wherein at least one R 6a is independently of each other H, oxo (=0), thioxo (=S), CH3, -CH2CH3, CHF2, CF3or cyclopropyl;

[0121] (3c) R6is

[0122] (4c) R6is -C 0-3 alkyl-P(=0)(R 61 )2, -C 0-3 alkyl-(N=)S(=0)(R 61 )2or -C 0-3 alkyl-S(=0)(=NR 62 )R 63 ; said R 61 , R 62and R 63 are each independently H, CH3, -CH2CH3, or cyclopropyl;

[0123] (5c) R6is

[0124] (6c) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, two R 61 and the P or S atom to which they are attached together form a heterocycloalkyl, said heterocycloalkyl is a 5-6 membered heterocycloalkyl, said 5-6 membered heterocycloalkyl is optionally substituted with 1, 2, 3, or 4 R 6b ; each R 6b is independently H, halogen, oxo (=O), OH, NH2, CN, C 1-3 alkyl, -L2-C 1-3 alkyl, -L2-C 3-5 cycloalkyl, or -L2-4-6 membered heterocycloalkyl; wherein L2 is -O-, -NH-, or -C(=O)-;

[0125] (7c) R6is ; said R 6b is hydrogen, L2-C 1-3 alkyl, L2-C 3-5 cycloalkyl, L2-4-6 membered heterocycloalkyl, wherein L2 is -C(=O)-;

[0126] (8c) R6is ; said R 6b is hydrogen,

[0127] (9c) R6is -C 0-3 alkyl-S(=O)(=NR 62 )R 63 ; said R 62 and R 63 together with the N and S atom to which they are attached form a 4-10 membered heterocycloalkyl, said 5-10 membered heterocycloalkyl is optionally substituted with 1, 2, 3, or 4 R 6b ; R6is

[0128] (10c) R6is

[0129] In an optional embodiment of the application, the above-mentioned compound is selected from the following structures:

[0130] said R1, R8 are as described in the present application, wherein L1and R6satisfy one or more of the following conditions:

[0131] (1c) L1is a single bond, -CH2- or -O-CH2-;

[0132] (2c) R6is 4-8 membered heterocycloalkyl, 5-8 membered heterocycloalkenyl or 5-6 membered heteroaryl, each independently optionally substituted with 1, 2, 3 or 4 R 6a ; each R 6a is independently H, oxo (=0), thioxo (C=S), CH3, CHF2or cyclopropyl;

[0133] (3c) R6is

[0134] (4c) R6is -C 0-3 alkyl-P(=0)(R 61 )2, -C 0-3 alkyl-(N=)S(=0)(R 61 )2or -C 0-3 alkyl-S(=0)(=NR 62 )R 63 ; each R 61 , R 62 and R 63 is independently H or CH3;

[0135] (5c) R6is

[0136] (6c) R6is -C 0-3 alkyl-P(=0)(R 61 )2, -C 0-3 alkyl-(N=)S(=0)(R 61 )2, two R 61 and the P or S atom to which they are attached together form a heterocycloalkyl, which is a 5-6 membered heterocycloalkyl, which is optionally substituted with 1, 2, 3 or 4 R 6b ; each R 6b is independently H, halogen, oxo (=0), OH, NH2, CN, C 1-3 alkyl, -L2-C 1-3 alkyl, -L2-C 3-5 cycloalkyl or -L2-4-6 membered heterocycloalkyl; wherein L2is -0-, -NH- or -C(=0)-;

[0137] (7c) R6is said R 6b is hydrogen, L2-C 1-3 alkyl, L2-C 3-5 cycloalkyl, L2-4-6 membered heterocycloalkyl, wherein L2is -C(=O)-;

[0138] (8c) R6is said R 6b is hydrogen,

[0139] (9c) R6is

[0140] In an optional embodiment of the application, the above compounds are selected from the following structures:

[0141] said L1and R8are as defined herein, wherein R6satisfies one or more of the following conditions:

[0142] (1d) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, or -C 0-3 alkyl-S(=O)(=NR 62 )R 63 ;

[0143] (2d) R6is

[0144] (3d) R6is 4-8 membered heterocycloalkyl or 5-6 membered heterocycloalkenyl, said 4-8 membered heterocycloalkyl and 5-6 membered heterocycloalkenyl being substituted with at least one thioxo (C=S);

[0145] (4d) R6is

[0146] In an optional embodiment of the application, the above compounds are selected from the following structures:

[0147] wherein R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, or -C 0-3 alkyl-S(=O)(=NR 62 )R 63 ;

[0148] and / or, R6is

[0149] and / or, R6is 5-6 membered heterocyclenyl, said 5-6 membered heterocyclenyl is substituted with at least one thio (C=S);

[0150] and / or, R6is said R8is as defined herein.

[0151] In an alternative embodiment of the application, L1is a single bond and R6is selected from

[0152] In an alternative embodiment of the application, R6is

[0153] In an alternative embodiment of the application, R6is

[0154] In an alternative embodiment of the application, R6is

[0155] In an alternative embodiment of the application, R6is

[0156] In an alternative embodiment of the application, R6is

[0157] In an alternative embodiment of the application, R6is

[0158] In an alternative embodiment of the application:

[0159] (1e) R6is 4-8 membered heterocycloalkyl, 5-8 membered heterocyclenyl or 5-6 membered heteroaryl, each independently optionally substituted with 1, 2, 3, 4 or 5 R 6a ; or,

[0160] (2e) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2 or -C 0-3 alkyl-S(=O)(=NR62 )R 63 , said R 61 , R 62 and R 63 are each independently H, C 1-6 alkyl or C 3-6 cycloalkyl; or,

[0161] (3e) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, said two R 61 and the P or S atom to which they are attached together form a 4-6 membered heterocycloalkyl group, said 4-6 membered heterocycloalkyl group being optionally substituted with 1, 2, 3 or 4 R 6b ; or,

[0162] (4e) R6is -C 0-3 alkyl-S(=O)(=NR 62 )R 63 , said R 62 and R 63 together with the N and S atom to which they are attached form a 5-10 membered heterocycloalkenyl group, said 5-10 membered heterocycloalkenyl group being optionally substituted with 1, 2, 3 or 4 R 6b ; or,

[0163] (5e) R6is a 4-8 membered heterocycloalkyl group or a 5-8 membered heterocycloalkenyl group, said 4-8 membered heterocycloalkyl group and 5-8 membered heterocycloalkenyl group being substituted with 1, 2, 3, 4, 5 or 6 R 6a , wherein at least one R 6a is thio (=S).

[0164] In an optional embodiment of the application:

[0165] (1f) R6is Preferably, R6is or,

[0166] (2f) R6is or,

[0167] (3f) R6is

[0168] Preferably, R6is or,

[0169] (4f) R6is or

[0170] (5f) R6 is

[0171] Preferably, R6 is

[0172] In an optional embodiment of the present application, the compound has the following structure:

[0173] R6 is selected from -(N=)S(=0)(R 61 )2, 4-7 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl; said R 61 each independently is CH3, CHF2, or cyclopropyl; said 4-7 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl is optionally substituted with 1, 2, 3, 4, or 5 R 6a substituents, wherein at least one R 6a is selected from thio(C=S); or, two R 61 and the S atom to which they are attached together form a 5-6 membered heterocycloalkyl, said 5-6 membered heterocycloalkyl is optionally substituted with 1, 2, 3, or 4 R 6b substituents;

[0174] R6 is selected from

[0175] Preferably, R6 is selected from

[0176] More preferably, R6 is selected from

[0177] In an optional embodiment of the present application, the compound has the following structure:

[0178] In a second aspect of the present application, there is provided a pharmaceutical composition comprising: a compound, a tautomer, a stereoisomer, a pharmaceutically acceptable salt, or a prodrug thereof as described in the first aspect of the present application; and a pharmaceutically acceptable excipient.

[0179] In a third aspect of the present application, there is provided use of a compound, a tautomer, a stereoisomer, a pharmaceutically acceptable salt, or a prodrug thereof as described in the first aspect of the present application, and a pharmaceutical composition as described in the second aspect of the present application in the manufacture of a medicament for treating or preventing a VAV1 -related disease.

[0180] In an optional embodiment of the present application, the VAV1 -related disease comprises cancer and autoimmune diseases.

[0181] In an optional embodiment of the present application, the above-mentioned medicament is used for treating or preventing cancer and autoimmune diseases.

[0182] In an optional embodiment of the present application, the above-mentioned VAV1 -related diseases include systemic lupus erythematosus, myasthenia gravis, periodontitis, type I diabetes, rheumatoid arthritis, multiple sclerosis, inflammatory enteritis, inflammatory bowel disease, autoimmune hepatitis, or psoriasis.

[0183] In an optional embodiment of the present application, the above-mentioned medicament is used for treating or preventing diseases including but not limited to systemic lupus erythematosus, myasthenia gravis, periodontitis, type I diabetes, rheumatoid arthritis, multiple sclerosis, inflammatory enteritis, inflammatory bowel disease, colitis, autoimmune hepatitis, or psoriasis.

[0184] In an optional embodiment of the present application, the above-mentioned VAV1 -related diseases include but are not limited to systemic lupus erythematosus, myasthenia gravis, periodontitis, type I diabetes, rheumatoid arthritis, multiple sclerosis, inflammatory enteritis, inflammatory bowel disease, autoimmune hepatitis, psoriasis, and the like.

[0185] Additional aspects and advantages of the application will be set forth in part in the description that follows, and in part will become apparent to those skilled in the art upon examination of the following description, or can be learned by practice of the application.

[0186] Terms and Definitions

[0187] Unless otherwise defined, all terms and definitions used in the present application, including the specification and claims, are as follows.

[0188] As will be understood by those skilled in the art, according to the convention used in the art, in the structural formulae of the present application, is used to depict a chemical bond, which is the point of attachment of a moiety or substituent to the core structure or backbone structure.

[0189] Unless otherwise specified, the term "pharmaceutically acceptable" in reference to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0190] Unless otherwise specified, the term "pharmaceutically acceptable salt" refers to a non-toxic acid or base salt of a pharmaceutically acceptable salt, including salts of inorganic acids and bases, salts of organic acids and bases.

[0191] In addition to pharmaceutically acceptable salts, other salts are also contemplated. They can serve as intermediates in the purification of the compounds or in the preparation of other pharmaceutically acceptable salts, or they can be useful in the identification, characterization, or purification of the compounds of the present application.

[0192] Unless otherwise specified, the term "pharmaceutical composition" means one or more of the compounds described herein or a physiologically / pharmaceutically acceptable salt or prodrug thereof in combination with a further chemical component, such as a physiologically / pharmaceutically acceptable carrier and excipient. The purpose of a pharmaceutical composition is to facilitate administration of the compound to an organism.

[0193] Unless otherwise specified, the term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of classes of excipients include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, and diluents. Excipients can enhance the handling properties of a pharmaceutical formulation, i.e., make the formulation more amenable to direct compression by increasing flow and / or cohesion.

[0194] Unless otherwise specified, the term "prodrug" refers to compounds that can be converted under physiological conditions or by solvolysis to a compound of the present application that is biologically or pharmaceutically active. Prodrugs of the present application are prepared by modifying functional groups in such a way that their solubility characteristics are altered but the biologically or pharmaceutically useful properties are retained. The prodrugs of the present application are prepared in accordance with conventional methods and either are known or will be known by one skilled in the art. Prodrugs include compounds wherein a hydroxy or amino group in a compound of the present application is linked to any group that can be cleaved in vivo to form the free hydroxyl or amino group. Prodrugs of the present application are cleaved when a prodrug of the present application is administered to a mammalian subject to form the free hydroxyl or amino group.

[0195] Unless otherwise specified, the term "stereoisomer" refers to isomers that have the same molecular formula but different structures resulting from the different spatial arrangement of atoms. Stereoisomers include enantiomers, diastereomers, and conformational isomers.

[0196] Depending on the choice of starting materials and methods, the compounds of the application can be present in the form of one or more of possible isomers, such as geometric or stereoisomers, for example as pure optical isomers, or as mixtures of different isomers, such as racemates, diastereomeric mixtures, and enantiomeric mixtures. When describing compounds having optical activity, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule with respect to a single reference plane. The prefixes D and L or (+) and (-) are used to designate the plane polarized light that the compound will cause to rotate. (-) or L means that the compound is levorotatory. A compound of the prefix (+) or D is dextrorotatory. Except for these stereoisomers, these compounds are identical and indistinguishable from one another. Specific stereoisomers are also referred to as enantiomers, and mixtures of such isomers are often referred to as enantiomeric mixtures. A 50:50 mixture of enantiomers is referred to as a racemic mixture or racemate, and such mixtures can occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. Many geometric isomers of olefins, C=N double bonds, and the like can also exist; all such stable isomers are contemplated in the present application. When the compounds described herein contain olefinic double bonds, unless otherwise specified, both E and Z geometric isomers are encompassed. If the compound contains a disubstituted cycloalkyl group, the substituents can be in the cis- or trans- (or, alternatively, the Z- or E-) configuration.

[0197] When bonds to a chiral carbon in the formulas of the application are drawn in a straight line, it is understood that both the (R) and (S) configurations of the chiral carbon, and the enantiomerically pure compounds and mixtures resulting therefrom, are included within the scope of the general formula. The graphical representation of racemates or enantiomerically pure compounds herein is from Maehr, J. Chem. Ed. 1985, 62:114-120. Unless otherwise indicated, a wedge and a dashed line represent the absolute configuration of one stereogenic center.

[0198] Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. Compounds of the application containing asymmetrically substituted carbon atoms can exist in optically active form or as racemic mixtures. Resolution of racemic mixtures into their individual components can be achieved by any of a number of methods known in the art. Exemplary methods include fractional recrystallization using chiral resolving acids, which are optically active, salt-forming organic acids. Suitable resolving agents for fractional recrystallization procedures are, for example, optically active acids, such as the D and L forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid or the various optically active camphorsulfonic acids. Other suitable resolving agents for fractional crystallization procedures include the stereoisomerically pure forms of a-methyl- benzylamine (e.g., the S and R forms or diastereomeric pure forms), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, and the like. Resolution of racemic mixtures can also be achieved by elution of a chromatography column packed with an optically active resolving agent (e.g., a dinitrobenzoylphenylglycine). High performance liquid chromatography (HPLC) can be employed as can supercritical fluid chromatography (SFC). The choice of the particular method and elution conditions, as well as the choice of the chromatography column, can be selected by one skilled in the art based upon the structure of the compound and the results of the test. Further, any enantiomer or diastereomer of a compound described herein can be obtained by stereospecific synthesis from an optically pure starting material or reagent of known configuration.

[0199] The term "tautomer" refers to isomers of a molecule that differ only in the position of a proton, except where otherwise specified. Compounds of the application can exhibit tautomerism. Tautomeric compounds can exist in two or more interconvertible forms. Proton- shifting tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium and attempts to isolate a single tautomer usually result in a mixture whose physical properties are consistent with a mixture of compounds. The position of the equilibrium depends on the intramolecular chemistry. For example, in many aliphatic aldehydes and ketones, such as acetaldehyde, the keto form predominates; in phenols, the enol form predominates. The present application includes all tautomeric forms of the compounds. For example may interconvert.

[0200] Unless otherwise indicated, a wedged solid line bond and a wedged dashed line bond indicate the absolute configuration at a stereocenter, a straight solid line bond and a straight dashed line bond indicate the relative configuration at a stereocenter.

[0201] The compounds of the present application can contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds can be labeled with radioactive isotopes, such as deuterium (2H), tritium (3H), iodine-125 (125I) or C-14 (14C). All isotopic variations of the compounds of the present application, whether radioactive or not, are encompassed within the scope of the present application.

[0202] The term "effective amount" or "therapeutically effective amount" with respect to a drug or pharmacological agent refers to a sufficient amount of the drug or agent to provide the desired effect, without being toxic to the subject. For oral dosage forms of the present application, an "effective amount" of one active agent in a composition refers to the amount needed to achieve the desired effect in conjunction with the other active agent in the composition. The determination of an effective amount is dependent on the age and general condition of the subject, as well as the particular active agent, and an appropriate effective amount for a given case can be determined by one of ordinary skill in the art using routine testing.

[0203] The term "active ingredient," "therapeutic agent," "active agent," or "active agent" refers to a chemical entity that is effective in treating a disorder, disease, or condition of interest, unless otherwise specified.

[0204] The term "substituted," unless otherwise specified, means that any one or more hydrogen atoms on the particular atom is / are replaced with a substituent group, including heavy isotopes and variations of hydrogen, as long as the valency of the particular atom is not exceeded and the substituted compound is stable. When the substituent is oxo (i.e., =0), it can be represented as oxidation of carbon, nitrogen, and sulfur atoms, including but not limited to C(=0), S(=0), S(=0)2, or N(=0).

[0205] The term "optionally" or "optional," unless otherwise specified, means that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0206] The term "optionally substituted" means that the group can or can not be substituted and that the types and number of substituents are optional on a chemical basis.

[0207] When any variable (e.g., R) occurs more than one time in a compound, its definition in each instance is independent of the definition of the other occurrences. Thus, for example, if a group is substituted with 0-2 R groups, then the group is optionally substituted with up to two R groups, and the R group is independently selected at each occurrence. Also, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0208] The term "D" is used to represent the isotope deuterium (2H) unless otherwise specified. 2 H).

[0209] The term "C 1-6 alkyl" is used to denote a straight or branched saturated hydrocarbon group consisting of from 1 to 6 carbon atoms. The C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C6and C5alkyl groups and the like; which can be monovalent (e.g., methyl), divalent (e.g., methylene) or multivalent (e.g., methine). Examples of C 1-6 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl and the like.

[0210] The term "C 1-3 alkyl" is used to denote a straight or branched saturated hydrocarbon group consisting of from 1 to 3 carbon atoms. The C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups and the like; which can be monovalent (e.g., methyl), divalent (e.g., methylene) or multivalent (e.g., methine). Examples of C 1-3 alkyl groups include, but are not limited to, methyl (Me), methylene (-CH2-), ethyl (Et), propyl (including n-propyl and isopropyl) and the like.

[0211] The term "halo" is used interchangeably with the term "halogen substituted" when used alone or as part of another substituent.

[0212] "Haloalkyl" or "halogen substituted alkyl" means, unless otherwise specified, a branched and straight chain saturated aliphatic hydrocarbon radical having the specified number of carbon atoms, substituted by one or more halogens.

[0213] "C 2-6 alkenyl" is used to denote a straight or branched hydrocarbon group consisting of from 2 to 6 carbon atoms containing at least one carbon-carbon double bond, which can be located in any position on the group. The C 2-6 alkenyl group includes C 2-4 , C 2-3 , C4, C3and C2alkenyl groups and the like; which can be monovalent, divalent or multivalent. Examples of C 2-6 alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl and the like.

[0214] Unless otherwise specified, "C 2-6 "Alkyne" is used to denote a straight-chain or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon triple bond, which can be located at any position within the group. 2-6 Alkyne groups include C 2-4 C 2-3 C4, C3, and C2 alkynyl groups, etc. They can be monovalent, divalent, or polyvalent. 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, and penynyl.

[0215] Unless otherwise specified, the term "C" 1-6 "Alkoxy" refers to alkyl groups containing 1 to 6 carbon atoms that are attached to the rest of the molecule by an oxygen atom. The C 1-6 Alkoxy groups include C 1-4 C 1-3 C 1-2 C 2-6 C 2-4 C6, C5, C4, and C3 alkoxy groups, etc. 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), pentoxy (including n-pentoxy, isopentoxy and neopentoxy), hexoxy, etc.

[0216] Unless otherwise specified, the term "C" 1-3 "Alkoxy" refers to alkyl groups containing 1 to 3 carbon atoms that are attached to the rest of the molecule by an oxygen atom. The C 1-3 Alkoxy groups include C 1-2 C 2-3 Examples of C1-3 alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy).

[0217] Unless otherwise specified, the term "C" 1-3 "Alkylamino" refers to alkyl groups containing 1 to 3 carbon atoms that are attached to the rest of the molecule via an amino group. The C 1-3 Alkylamino groups include C 1-2 C3 and C2 alkylamino groups, etc. C 1-3 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -NHCH2CH2CH3, -NHCHCH3)2, etc.

[0218] Unless otherwise specified, the term "C"3-12 Cycloalkyl" denotes a saturated cyclic hydrocarbyl group consisting of 3 to 12 carbon atoms, including monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spiro, fused, and bridged rings. The C 3-12 Cycloalkyl includes C 3-10 , C 3-8 , C 3-6 , C 3-5 , C 4-8 , C 4- 6, C 4-5 , C 5-8 or C 5-6 cycloalkyl and the like; which can be monovalent, divalent, or multivalent. C 3-8 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [2.2.2]bicyclooctane, and the like.

[0219] Unless otherwise specified, the term "C 3-8 Cycloalkyl" denotes a saturated cyclic hydrocarbyl group consisting of 3 to 8 carbon atoms, including monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spiro, fused, and bridged rings. The C 3-8 Cycloalkyl includes C 3-6 , C 3-5 , C 4-8 , C 4-6 , C 4-5 , C 5-8 or C 5-6 cycloalkyl and the like; which can be monovalent, divalent, or multivalent. C 3-8 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [2.2.2]bicyclooctane, and the like.

[0220] Unless otherwise specified, the term "C 3-6 Cycloalkyl" denotes a saturated cyclic hydrocarbyl group consisting of 3 to 6 carbon atoms, which is monocyclic and bicyclic ring systems, the C 3-6 Cycloalkyl includes C 3-5 , C 4-5 and C 5-6 cycloalkyl and the like; which can be monovalent, divalent, or multivalent. C 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0221] Unless otherwise specified, Cn-n+m or Cn-Cn+m includes any one particular instance of n to n+m carbons, for example C 1-12 includes C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12Also included are any range within n to n+m, for example C 1-12 Also included are any range within n to n+m, for example C 1- 3, C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12 etc. Likewise, n-member to n+m-member means the number of atoms in the ring is n to n+m, for example 3-12 membered ring includes 3 membered, 4 membered, 5 membered, 6 membered, 7 membered, 8 membered, 9 membered, 10 membered, 11 membered, and 12 membered rings, also including any range within n to n+m, for example 3-12 membered ring includes 3-6 membered ring, 3-9 membered ring, 5-6 membered ring, 5-7 membered ring, 6-7 membered ring, 6-8 membered ring, and 6-10 membered ring, etc.

[0222] The term "heterocycloalkyl" when used alone or as part of another substituent group refers to a cycloalkyl group in which one or more (in some embodiments 1 to 3) carbon atoms are replaced by a heteroatom such as, but not limited to, N, NH, O, S, P, S(=0), S(=0)2, or S(=0)(=NH). The term "m-n membered heterocycloalkyl" is understood to mean a saturated ring having m to n atoms, wherein the hetero ring atoms are selected from N, O, S, P, preferably from N, O, or S. For example, the term "4-8 membered heterocycloalkyl" is understood to mean a saturated or partially saturated ring having 4 to 8 atoms, wherein 1, 2, 3, or 4 ring atoms are selected from N, NH, O, S, P, S(=0), S(=0)2, or S(=0)(=NH), preferably from N, O, or S. A "4-10 membered heterocyclyl" is then understood to mean a saturated ring having 4 to 10 atoms. When a prefix such as 4-8 membered or 4-10 membered is used to indicate a heterocycloalkyl group, the number of carbons also means to include the heteroatoms. Single, bi-, tri-, spiro, or bridged rings are included. Examples of heterocycloalkyl groups are: pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydropyridinyl, tetrahydropyrrolyl, azetidinyl, thiazolidinyl, oxazolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, azepanyl, diazepanyl, oxazepanyl, etc. The term "heterocycloalkyl" can be used interchangeably with the term "heteroalkyl ring".

[0223] The term "aromatic ring," by itself or in combination with other terms, means a monocyclic or polycyclic carbocyclic ring having from 6 to 20 carbon atoms, wherein at least one ring is aromatic. When one of the rings is non-aromatic, the group can be attached through an aromatic ring or through a non-aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl, and acenaphthyl. The term "aromatic ring" can be used interchangeably with the term "aryl."

[0224] The term "heteroaromatic ring," by itself or in combination with other terms, means a monocyclic or polycyclic carbocyclic ring wherein at least one ring atom is a heteroatom independently selected from oxygen, sulfur, and nitrogen, the remainder of the ring atoms being carbon, wherein at least one ring is aromatic. The group can be carbon-based or heteroatom-based (i.e., it can be C-attached or N-attached, as is possible). When one of the rings is non-aromatic, the group can be attached through an aromatic ring or through a non-aromatic ring. Examples of heteroaryl groups include, but are not limited to, imidazolyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, N-methylpyrrolyl, and tetrahydroquinoline. The term "heteroaromatic ring" can be used interchangeably with the terms "heteroaromatic ring," "heteroaryl," or "heteroaromatic group."

[0225] The term "5-6 membered heterocyclenyl," by itself or in combination with other terms, means an unsaturated or partially unsaturated cyclic group consisting of 5 to 6 ring atoms, but not an aromatic ring, 1, 2, 3, or 4 of which are heteroatoms independently selected from O, S, and N, the remainder of which are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen, and sulfur heteroatoms are optionally oxidized (i.e., C(=O), NO, and S(O)p, where p is 1 or 2), unless otherwise specified. In addition, for this "5-6 membered heterocyclenyl," a heteroatom can occupy the position of attachment of the heterocyclenyl group to the rest of the molecule. The 5-6 membered heterocyclenyl group includes 5-membered and 6-membered heterocyclenyl groups, and the like.

[0226] Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" are used interchangeably throughout the application, and the term "5-6 membered heteroaryl" means a monocyclic ring group consisting of 5 to 6 ring atoms having a conjugated pi-electron system, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the remainder carbon atoms. Of the nitrogen atoms, one can optionally be quaternized, and the nitrogen and sulfur heteroatoms can optionally be oxidized (i.e., NO and S(O)p, where p is 1 or 2). The 5-6 membered heteroaryl can be attached to the remainder of the molecule by a heteroatom or carbon atom. The 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryls. Examples of the 5-6 membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1, 2, 3-triazolyl, 2H-1, 2, 3-triazolyl, 1H-1, 2, 4-triazolyl and 4H-1, 2, 4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thiophenyl (including 2-thiophenyl and 3-thiophenyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

[0227] Unless otherwise specified, the term "halo" or "halogen" is fluorine, chlorine, bromine and iodine.

[0228] Further, it is to be noted that the description manner "independently" employed in the present application should be interpreted in a broad sense, unless explicitly indicated otherwise, that is, each individual described is independent of each other, and can be independently the same or different specific group. In more detail, the description manner "independently" can mean that in different groups, the specific options expressed between the same symbols do not affect each other, or it can mean that in the same group, the specific options expressed between the same symbols do not affect each other.

[0229] Unless otherwise specified, the term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine; cattle, sheep, horses, or primates, and most preferably humans.

[0230] The term "therapeutically effective amount" means, unless otherwise indicated, an amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response that is being sought in a tissue, system, animal, individual or human by a researcher, veterinarian, medical doctor or other clinician, which includes one or more of: (1) preventing the disease: for example, preventing a disease, disorder or condition from occurring in an individual that is predisposed to the disease, disorder or condition but has not yet been diagnosed with the disease pathology or symptoms; (2) inhibiting the disease: for example, arresting the development of a disease, disorder or condition in an individual that is experiencing or displaying the pathology or symptoms of the disease, disorder or condition (i.e., halting the pathology and / or symptoms); (3) relieving the disease: for example, causing the regression of the pathology and / or symptoms in an individual that is experiencing or displaying the pathology or symptoms of the disease, disorder or condition (i.e., reversing the pathology and / or symptoms).

[0231] The terms "treat" and other similar synonymous terms as used herein include the following meanings:

[0232] (i) preventing the disease or condition from occurring in a mammal, in particular, when such mammal is predisposed or has a predisposition to the disease or condition but has not yet been diagnosed as having it;

[0233] (ii) inhibiting the disease or condition, i.e., arresting its development;

[0234] (iii) relieving the disease or condition, i.e., causing the state of the disease or condition to regress; or

[0235] (iv) alleviating the symptoms of the disease or condition.

[0236] Abbreviations used in the present invention are defined as follows:

[0237] DC 50 : half maximal effective concentration, the concentration of a compound that brings a response halfway between the baseline and maximum at a given assay

[0238] M: molar concentration, e.g., 1 M hydrochloric acid means 1 mol / L hydrochloric acid solution

[0239] N: normality, e.g., 2 N hydrochloric acid means 2 mol / L hydrochloric acid solution

[0240] DMSO: dimethyl sulfoxide

[0241] Bpin: pinacol boronate. Beneficial effects

[0242] According to the embodiments of the present application, the present application has at least one of the following technical effects:

[0243] 1) The compound of the present application can induce the direct binding of VAV1 and CRBN in a dose-dependent manner;

[0244] 2) The compound of the present application shows excellent degradation effect on VAV1 protein in a dose-dependent manner;

[0245] 3) In CD3 / CD28 induced Jurkat cell and T cell activation model, the compound of the present application can significantly inhibit the production of IL-2, and the inhibition function is positively correlated with the dose;

[0246] 4) The compound of the present application shows excellent pharmacokinetic properties in mouse and rat pharmacokinetic tests, and has good drug properties;

[0247] 5) The compound of the present application shows excellent human liver microsomal stability, good thermodynamic solubility, no obvious CYP3A4 inhibition effect, and significant advantage in plasma protein binding rate, and has good drug properties;

[0248] 6) The test compound group of the present application shows significant alleviation of disease progression in the mouse intestinal inflammation model induced by adoptive naive T cells. DETAILED DESCRIPTION

[0249] The present application will be further described in conjunction with specific examples. It should be understood that the following description is only the most preferred embodiment of the present application, and should not be considered as a limitation on the scope of protection of the present application. On the basis of a full understanding of the present application, the experimental methods not specified in the following examples are generally carried out under conventional conditions, or under the conditions recommended by the manufacturer, and the skilled person in the art can make non-essential changes to the technical solutions of the present application. Such changes should be considered to be within the scope of protection of the present application.

[0250] Preparation of intermediate A

[0251] 3-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)phenyl)piperidine-2,6-dione

[0252] The synthesis route of intermediate A is as follows:

[0253] First step: 2-(3-bromo-2-chlorophenyl)acetonitrile (A2)

[0254] Into a reaction flask was placed 3-bromo-2-chlorobenzyl bromide (Al) (9.0 g, 31.65 mmol), acetonitrile (100 mL), trimethylsilyl cyanide (6.3 g, 63.3 mmol), and potassium carbonate (13 g, 100 mmol). The reaction was stirred at 80 °C for 16 h. After the reaction was completed, the reaction was cooled to room temperature, diluted with ethyl acetate (500 mL), washed with saturated aqueous sodium chloride solution (500 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V / V) = 10: 1) to give 2-(3-bromo-2-chlorophenyl)acetonitrile (A2).

[0255] LC-MS, M / Z (ESI): 230.1 [M+H] + .

[0256] Second step: methyl 4-(3-bromo-2-chlorophenyl)-4-cyanobutanoate (A3)

[0257] Into a reaction flask was placed 2-(3-bromo-2-chlorophenyl)acetonitrile (A2) (5.0 g, 21.74 mmol), methyl acrylate (3.74 g, 43.5 mmol), and tetrahydrofuran (100 mL), followed by sodium methoxide (110 mg, 2 mmol) under ice bath. The reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction was diluted with ethyl acetate (500 mL), washed with aqueous sodium chloride solution (500 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V / V) = 5: 1) to give methyl 4-(3-bromo-2-chlorophenyl)-4-cyanobutanoate (A3) (5.95 g, yield: 87%).

[0258] LC-MS, M / Z (ESI): 316.2 [M+H] + .

[0259] Third step: 3-(3-bromo-2-chlorophenyl)piperidine-2,6-dione (A4)

[0260] Methyl 4-(3-bromo-2-chlorophenyl)-4-cyanobutanoate (A3) (5.9 g, 18.7 mmol) was dissolved in acetic acid (100 mL), 0.5 mL of concentrated sulfuric acid was added at 20 °C, and the reaction was stirred at 90 °C for 6 h. After the reaction was completed, 2.0 g of sodium acetate was added at 20 °C, and stirred at room temperature for 30 min. The solvent was removed by distillation under reduced pressure, then diluted with ethyl acetate (500 mL), washed with saturated sodium chloride aqueous solution (500 mL x 1), saturated sodium bicarbonate aqueous solution (500 mL x 2), saturated sodium chloride aqueous solution (500 mL x 1), respectively, then the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 1:1) to give compound 3-(3-bromo-2-chlorophenyl)piperidine-2,6-dione (A4).

[0261] LC-MS, M / Z (ESI): 302.1 [M+H] + .

[0262] 1 H NMR (400 MHz, DMSO d6) δ 10.93 (s, 1H), 7.71 (dd, 1H), 7.37 (dd, 1H), 7.27 (t, 1H), 4.31 (dd, 1H), 2.77 (ddd, 1H), 2.58 - 2.49 (m, 1H), 2.31 (qd, 1H), 2.04 - 1.95 (m, 1H).

[0263] Fourth step: 3-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine- 2,6-dione

[0264] Compound 3-(3-bromo-2-chlorophenyl)piperidine-2,6-dione (A4) (4.6 g, 15.2 mmol) was placed in a reaction flask, then pinacol diboronic acid (7.6 g, 30 mmol), potassium acetate (4.5 g, 45.6 mmol) and 1,1'-bis(di-phenylphosphino)ferrocene palladium chloride (0.3 g, 0.45 mmol), 1,4-dioxane (70 mL) were added, and the reaction was stirred at 100 °C for 8 h under nitrogen protection. The reaction was cooled to room temperature, diluted with ethyl acetate (500 mL), washed with saturated sodium chloride aqueous solution (500 mL x 3), then the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 1:1) to give compound 3-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine-2,6-dione (intermediate A).

[0265] LC-MS, M / Z (ESI): 350.2 [M+H] + .

[0266] Example 1: Preparation of target compound 1

[0267] 3-[2-chloro-4'-(dimethylphosphoryl)[1,1'-biphenyl]-3-yl]piperidine-2,6-dione (Compound 1)

[0268] The synthetic route of target compound 1 is shown as follows:

[0269] First step: Synthesis of (4-bromophenyl)bis(methyl)(oxo)-λ 5 - phosphane (Compound 1-2)

[0270] Dissolve 4-bromo-iodobenzene (1.0 g, 3.5 mmol), dimethylphosphine oxide (900 mg, 10.5 mmol), triethylamine (1.8 g, 17.5 mmol), tris(dibenzylideneacetone)dipalladium (300 mg, 0.3 mmol), 4,5-bisdiphenylphosphino-9,9-dimethylxanthene (350 mg, 0.6 mmol) in 1,4-dioxane (15 mL), and stir the reaction solution at 60°C under nitrogen protection overnight. After the reaction is complete, dilute the reaction solution with saturated brine (60 mL), then extract with ethyl acetate (50 mL x 2), dry the combined organic phases over sodium sulfate, filter, and concentrate to obtain a crude product, which is purified by column chromatography (petroleum ether / ethyl acetate (V / V) = 1 / 1) to obtain (4-bromophenyl)bis(methyl)(oxo)-λ 5 - phosphane (Compound 1-2).

[0271] LC-MS, M / Z (ESI): 233.2 / 235.2 [M+H] +

[0272] Second step: Synthesis of dimethyl(oxo)[4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)phenyl]-λ 5 - phosphane (Compound 1-3)

[0273] Dissolve (4-bromophenyl)bis(methyl)(oxo)-λ 5- phosphine (compound 1-2) (250 mg, 1.0 mmol), bis(pinacolato)diboron (893 mg, 3.5 mmol), potassium acetate (330 mg, 3.5 mmol) and l,l'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (40 mg, 0.07 mmol) were dissolved in anhydrous 1,4-dioxane (5 mL) and the reaction was stirred at 60 °C for 2 h under nitrogen. The reaction was dissolved in ethyl acetate (50 mL) and washed with saturated brine (10 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product which was purified by column chromatography (petroleum ether / ethyl acetate (V / V) = 1 / 1) to give compound dimethyl(oxo)[4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl]-λ 5 - phosphine (compound 1-3).

[0274] LC-MS, M / Z (ESI): 281.2 [M+H]+

[0275] Third step: synthesis of 3-[2-chloro-4'-(dimethylphosphoryl)[l,l'-biphenyl]-3- yl]piperidine-2,6-dione (compound 1)

[0276] 3-(3-bromo-2-chlorophenyl)piperidine-2,6-dione (intermediate A4) (70 mg, 0.25 mmol), dimethyl(oxo)[4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl]-λ 5 - phosphine (compound 1-3) (100 mg, 0.36 mmol), l,l'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (31 mg, 0.04 mmol), potassium phosphate (88 mg, 0.82 mmol) were dissolved in anhydrous N,N-dimethylformamide (1.5 mL) and the reaction was stirred at 100 °C for 4 h. After the reaction was completed, the reaction was cooled to room temperature and filtered. The mother liquor was purified by preparative liquid chromatography (column: Phenomenex Synergi C18 100*25 mm*4 μm; solvent: A = water + 0.1 volume% formic acid (99%), B = acetonitrile; gradient: 5%-95% in 7 minutes) to give compound 3-[2-chloro-4'-(dimethylphosphoryl)[l,l'-biphenyl]-3-yl]piperidine-2,6-dione (compound 1).

[0277] LC-MS, M / Z (ESI): 376.2 [M+H] +

[0278] 1H NMR (400 MHz, DMSO-d6) δ 10.93 (s, 1H), 7.86-7.30 (m, 7H), 4.35-4.31 (m, 1H), 2.79-1.65 (m, 10H).

[0279] Example 2: Preparation of compound 2

[0280] 3-(2-chloro-4'-((dimethyl(oxo)-λ 6 -sulfoxylidene)amino)-[1,1'-biphenyl]-3-yl)piperidine-2,6-dione (compound 2)

[0281] The synthetic route of the target compound 2 is shown as follows:

[0282] First step: synthesis of compound ((4-bromophenyl)imino)dimethyl-λ 6 -sulfoxonium (compound 2-1)

[0283] Compound p-bromoiodobenzene (600.00 mg, 2.12 mmol) and dimethyl sulfoximine (237.06 mg, 2.54 mmol) were dissolved in anhydrous 1,4-dioxane solution (6 mL) at room temperature, then cesium carbonate (967.41 mg, 2.97 mmol), tris(dibenzylideneacetone)dipalladium (58.26 mg, 0.06 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (72.25 mg, 0.12 mmol) were added, and the reaction solution was stirred at 110°C under nitrogen protection for 12 hours. TLC monitoring showed that the raw material was completely reacted, the stirring was stopped, the reaction solution was reduced to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (10 mL x 5), the organic phase was collected, dried with anhydrous sodium sulfate, the organic phase was concentrated by reduced pressure distillation, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 100:50) to obtain compound ((4-bromophenyl)imino)dimethyl-λ 6 -sulfoxonium (compound 2-1).

[0284] LC-MS, M / Z (ESI): 249.9 [M+H] +

[0285] Second step: synthesis of compound dimethyl((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)imino)-λ 6 -sulfoxonium (compound 2-

[0286] 2).

[0287] At room temperature, the compound ((4-bromophenyl)imino)dimethyl-λ 6 Thione (compound 2-1) (300 mg, 1.21 mmol) and pinacol diboronate (460.52 mg, 1.81 mmol) were dissolved in anhydrous 1,4-dioxane solution (5 mL), and potassium acetate (355.95 mg, 3.62 mmol) and 1,1-bis(diphenylphosphine)diferropalladium dichloride (88.46 mg, 0.12 mmol) were added. The reaction solution was stirred in an oil bath at 90 °C for 12 hours under nitrogen protection. After the reaction of the reactants was complete as monitored by TLC, stirring was stopped, the reaction solution was cooled to room temperature, and then diluted with water (20 mL). Extraction was performed with ethyl acetate (10 mL × 5), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane:methanol (V / V) = 100:3) to give the compound dimethyl((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)imino)-λ 6 -Thione (compound 2-2).

[0288] LC-MS, M / Z (ESI): 296.0 [M+H] +

[0289] Step 3: Compound 3-(2-chloro-4'-((dimethyl(oxo)-λ) 6 Synthesis of (-sulfoxide)amino)-[1,1'-biphenyl]-3-yl)piperidine-2,6-dione (compound 2)

[0290] At room temperature, the compound dimethyl((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)imino)-λ 6 -Thione (compound 2-2) (150.00 mg, 0.51 mmol) and compound 3-(3-bromo-2-chlorophenyl)piperidine-2,6-dione (intermediate A4) (153.73 mg, 0.51 mmol) were dissolved in anhydrous N,N-dimethylformamide (2 mL), potassium phosphate (323.58 mg, 1.52 mmol) and 1,1-bis(diphenylphosphine)diferro-palladium dichloride (37.18 mg, 0.05 mmol) were added, and the reaction solution was stirred at 100 °C for 12 hours under nitrogen protection. After the reaction of the starting materials was complete as monitored by TLC, stirring was stopped, the reaction solution was cooled to room temperature, and then diluted with water (15 mL). Extraction was performed with ethyl acetate (5 mL × 5), the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 100:50) to give compound 3-(2-chloro-4'-((dimethyl(oxo)-λ)).6 -[(4'-((S-methylsulfinyl)amino)-[1,1'-biphenyl]-3-yl)amino]-[1,1'-biphenyl]-3-yl)- piperidine-2,6-dione (Compound 2).

[0291] LC-MS, M / Z (ESI): 391.0 [M+H] +

[0292] 1 H NMR (400 MHz, DMSO-d6) δ 10.92 (s, 1H), 7.39 - 7.31 (m, 1H), 7.31 - 7.26 (m, 2H), 7.26 - 7.21 (m, 2H), 7.03 - 6.95 (m, 2H), 4.33 (dd, 1H), 3.26 (s, 6H), 2.78 (ddd, 1H), 2.56 (t, 1H), 2.32 (qd, 1H), 2.10 - 1.96 (m, 1H).

[0293] Example 3: Preparation of the target compound 3

[0294] 3-(2-chloro-4'-((S-methylsulfinyl)amino)methyl)-[1,1'-biphenyl]-3-yl)piperidine-2,6- dione (Compound 3)

[0295] The synthetic route of Compound 3 is shown as follows:

[0296] First step: synthesis of (4-bromobenzyl)(methyl)sulfane (Compound 3-2)

[0297] Put 1-bromo-4-(bromomethyl)benzene (3-1) (3.0 g, 12.09 mmol) in a reaction bottle, add ethanol (100 mL), and add sodium methanethiolate (4.2 g, 60 mmol) under ice bath. Stir the reaction solution under reflux condition for 16 h. After completion, cool the reaction solution to room temperature, concentrate under reduced pressure, and purify the residue by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 10:1) to obtain (4-bromobenzyl)(methyl)sulfane (Compound 3-2).

[0298] LC-MS, M / Z (ESI): 217.0 [M+H] + .

[0299] Second step: synthesis of (4-bromobenzyl)(imino)(methyl)-λ 6 -sulfone (3-3)

[0300] Intermediate (4-bromobenzyl)(methyl)sulfane (3-2) (2.2 g, 10.2 mmol), iodobenzene diacetate (8.1 g, 25 mmol), ammonium carbonate (3.8 g, 40 mmol) were dissolved in methanol (100 mL) and the reaction was stirred at room temperature for 2 h under nitrogen. After completion, the reaction was diluted with ethyl acetate (500 mL), washed with saturated aqueous sodium chloride (500 mL x 3), then the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 1:2) to give compound (4-bromobenzyl)(imino)(methyl)-λ 66 sulfone (3-3) (0.62 g, yield: 25%).

[0301] LC-MS, M / Z (ESI): 248.1 [M+H] + .

[0302] Step 3: Synthesis of 3-(2-chloro-4'-((S-methylsulfonylimino)methyl)-[1,1'-biphenyl]-3- yl)piperidine-2,6-dione (Compound 3)

[0303] Intermediate (4-bromobenzyl)(imino)(methyl)-λ 6 sulfone (3-3) (100 mg, 0.40 mmol) was placed in a reaction flask, followed by the addition of intermediate 3-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine- 2,6-dione (Intermediate A) (205 mg, 0.60 mmol), potassium phosphate (255 mg, 1.20 mmol) and 1,1'-bis(di-phenylphosphino)ferrocene palladium chloride (30 mg, 0.04 mmol) and 1,4-dioxane (4 mL). The reaction was stirred at 100 °C for 8 h under nitrogen. The reaction was cooled to room temperature, diluted with ethyl acetate (100 mL), washed with saturated aqueous sodium chloride (100 mL x 3), then the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (dichloromethane:methanol (V / V) = 20:1) to give compound 3-(2-chloro-4'-((S-methylsulfonylimino)methyl)-[1,1'-biphenyl]-3-yl)piperidine-2,6-dione (Compound 3) (18 mg, yield: 12%).

[0304] LC-MS, M / Z (ESI): 391.1 [M+H] + .

[0305] 1H NMR (400 MHz, DMSO-d6) δ 10.90 (s, 1H), 7.49 (d, 2H), 7.42 - 7.36 (m, 3H), 7.34 (dd, 1H), 7.29 (dd, 1H), 4.47 - 4.35 (m, 2H), 4.32 (dd, 1H), 3.66 (s, 1H), 2.84 - 2.67 (m, 4H), 2.56 - 2.47 (m, 1H), 2.39 - 2.22 (m, 1H), 2.07 - 1.96 (m, 1H).

[0306] Example 4: Preparation of the target compound 4

[0307] 3-(2-chloro-4'-(2-thioxopyridin-1(2H)-yl)-[1,1'-biphenyl]-3-yl)piperidine-2,6-dione

[0308] The synthetic route of compound 4 is shown as follows:

[0309] First step: 1-(4-bromophenyl)pyridine-2(1H)-thione (compound 4-2)

[0310] 1-(4-bromophenyl)pyridine-2(1H)-one (compound 4-1) (300 mg, 1.20 mmol) was placed in a reaction bottle, then Lawesson's reagent (1.0 g, 2.4 mmol), toluene (10 mL) were added, and the reaction was refluxed at 110°C under nitrogen protection for 2 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate (300 mL), extracted with saturated aqueous sodium chloride solution (300 mL x 3), then the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol (V / V) = 20:1) to obtain compound 1-(4-bromophenyl)pyridine-2(1H)-thione (compound 4-2).

[0311] LC-MS, M / Z (ESI): 265.9 [M+H] + .

[0312] Second step: 3-(2-chloro-4'-(2-thioxopyridin-1(2H)-yl)-[1,1'-biphenyl]-3-yl)piperidine-2,6-dione (compound 4)

[0313] Intermediate 1-(4-bromophenyl)pyridine-2(1H)-thione (4-2) (120 mg, 0.45 mmol) was placed in a reaction flask, followed by the addition of 3-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine-2,6-dione (Intermediate A) (210 mg, 0.60 mmol), potassium phosphate (286 mg, 1.35 mmol), 1,1'-bis(di-phenylphosphino)ferrocene palladium chloride (36 mg, 0.05 mmol) and 1,4-dioxane (4 mL). After the addition was completed, the reaction was stirred at 100 °C for 6 hours under nitrogen protection. The reaction was cooled to room temperature, then the reaction was diluted with ethyl acetate (200 mL), washed with saturated aqueous sodium chloride solution (200 mL x 3), then the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane:methanol (V / V) = 10:1) to obtain compound 3-(2-chloro-4'-(2-thioxopyridin-1(2H)-yl)-[1,1'-biphenyl]-3-yl)piperidine-2,6-dione (compound 4).

[0314] LC-MS, M / Z (ESI): 409.0 [M+H] + .

[0315] 1 H NMR (400 MHz, DMSO-d6) δ 10.93 (s, 1H), 8.06-8.02 (m, 1H), 7.60-7.51 (m, 3H), 7.51-7.28 (m, 6H), 6.85 (td, 1H), 4.42-4.32 (m, 1H), 2.85-2.73 (m, 1H), 2.54 (dt, 1H), 2.34 (ddd, 1H), 2.10-2.01 (m, 1H).

[0316] Example 5: Synthesis of compound 5

[0317] 3-{2-chloro-4'-[1-methyl-5-sulfanyl-3-(trifluoromethyl)-1,5-dihydro-4H-1,2,4-triazol-4-yl][1,1'-biphenyl]-3-yl}piperidine-2,6-dione

[0318] The synthetic route of compound 5 is as follows:

[0319] First step: 2-[(4-bromophenyl)aminosulfonyl]-2-methylhydrazine-1-carboxylic acid tert-butyl ester (compound 5-3)

[0320] Compound 5-3) was obtained by dissolving 2-methylhydrazine-1-carboxylic acid tert-butyl ester (5-2) (303 mg, 2.07 mmol) in anhydrous dichloromethane (10 mL) at room temperature, then slowly adding 1-bromo-4-isothiocyanatobenzene (5-1) (500 mg, 1.73 mmol) at 0 °C, and stirring the reaction solution at 25 °C for 12 hours after the addition was completed. After the reaction was completed, the reaction solution was concentrated, and the obtained crude product was purified by column chromatography (petroleum ether: ethyl acetate (V / V) = 100:1 to 1:1) to obtain 2-[(4-bromophenyl)aminothioyl]-2-methylhydrazine-1-carboxylic acid tert-butyl ester (compound 5-3).

[0321] LC / MS (ESI) (m / z): 360.0 (M+H) + .

[0322] Second step: N-(4-bromophenyl)-1-methylhydrazine-1-thioamide (compound 5-4)

[0323] Compound 5-3) was obtained by dissolving 2-methylhydrazine-1-carboxylic acid tert-butyl ester (5-2) (303 mg, 2.07 mmol) in anhydrous dichloromethane (10 mL) at room temperature, then slowly adding 1-bromo-4-isothiocyanatobenzene (5-1) (500 mg, 1.73 mmol) at 0 °C, and stirring the reaction solution at 25 °C for 12 hours after the addition was completed. After the reaction was completed, the reaction solution was concentrated, and the obtained crude product was purified by column chromatography (petroleum ether: ethyl acetate (V / V) = 100:1 to 1:1) to obtain 2-[(4-bromophenyl)aminothioyl]-2-methylhydrazine-1-carboxylic acid tert-butyl ester (compound 5-3).

[0324] LC / MS (ESI) (m / z): 260 (M+H) + .

[0325] Third step: 4-(4-bromophenyl)-2-methyl-5-(trifluoromethyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (compound 5-5)

[0326] Compound 5-3) was obtained by dissolving 2-methylhydrazine-1-carboxylic acid tert-butyl ester (5-2) (303 mg, 2.07 mmol) in anhydrous dichloromethane (10 mL) at room temperature, then slowly adding 1-bromo-4-isothiocyanatobenzene (5-1) (500 mg, 1.73 mmol) at 0 °C, and stirring the reaction solution at 25 °C for 12 hours after the addition was completed. After the reaction was completed, the reaction solution was concentrated, and the obtained crude product was purified by column chromatography (petroleum ether: ethyl acetate (V / V) = 100:1 to 1:1) to obtain 2-[(4-bromophenyl)aminothioyl]-2-methylhydrazine-1-carboxylic acid tert-butyl ester (compound 5-3).

[0327] LC / MS (ESI) (m / z): 339.0 (M+H) + .

[0328] Fourth Step: 3-{2-Chloro-4'-[1-methyl-5-sulfanyl-3-(trifluoromethyl)-1,5-dihydro-4H-1,2,4-triazol-4-yl][1,1'-biphenyl]-3-yl}piperidine-2,6-dione (Compound 5)

[0329] Fourth Step: 3-{2-Chloro-4'-[1-methyl-5-sulfanyl-3-(trifluoromethyl)-1,5-dihydro-4H-1,2,4-triazol-4-yl][1,1'-biphenyl]-3-yl}piperidine-2,6-dione (Compound 5)

[0330] LC / MS (ESI) (m / z): 481.0 (M+H) + ;

[0331] 1 H NMR (400 MHz, DMSO-d6) δ 10.93 (s, 1H), 7.60 (q, 4H), 7.43 - 7.37 (m, 3H), 4.35 (dd, 1H), 3.82 (s, 3H), 2.82 - 2.74 (m, 1H), 2.55 - 2.49 (m, 1H), 2.32 (dd, 1H), 2.06 - 2.01 (m, 1H).

[0332] Example 6: Preparation of the target compound 6

[0333] 3-{2-Chloro-4'-[(1-oxo-1λ6 - (thiacyclopropan-1 -ylidene) amino] [1,1 '-biphenyl]-3-yl} piperidine-2, 6-dione (Compound 6)

[0334] The synthetic route of the target compound 6 is shown as follows:

[0335] First step: 1-[(4-bromophenyl)imino]-1λ 6 - Synthesis of thiacyclopentan-1-one (6-2)

[0336] 4-bromo-iodobenzene (800 mg, 2.8 mmol), 1-imino-1λ 6 - thiacyclopentan-1-one (6-1) (388 mg, 3.2 mmol), cesium carbonate (1.4 g, 4.2 mmol), tris(dibenzylideneacetone)dipalladium (260 mg, 0.3 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (350 mg, 0.6 mmol) were dissolved in 1,4-dioxane (10 mL), the reaction was stirred at 100 °C for 5 h under nitrogen protection. After the reaction was completed, the reaction was cooled to room temperature, diluted with a brine solution (60 mL), then extracted with ethyl acetate (50 mL*2), the organic phase was combined, dried over sodium sulfate, concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether / ethyl acetate (V / V) = 3 / 1) to obtain 1-[(4-bromophenyl)imino]-1λ 6 - thiacyclopentan-1-one (6-2).

[0337] LC-MS, M / Z (ESI): 274.2 / 276.2 [M+H]+

[0338] Second step: 1-{[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]imino}-1λ 6 - Synthesis of thiacyclopentan-1-one (6-3)

[0339] 1-[(4-bromophenyl)imino]-1λ 6Thionitropentan-1-one (6-2) (200 mg, 0.7 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis-1,3,2-dioxaborane (893 mg, 3.5 mmol), potassium acetate (330 mg, 3.5 mmol), and 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (II) (40 mg, 0.07 mmol) were dissolved in anhydrous 1,4-dioxane (5 mL). The reaction mixture was stirred at 60 °C for 2 h under nitrogen protection. The reaction solution was cooled to room temperature, and ethyl acetate (20 mL) was added. The mixture was then washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (petroleum ether / ethyl acetate (V / V) = 2 / 1) to give compound 1-{[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl]imino}-1λ 6 -Thiocyclopentane-1-one (6-3).

[0340] LC-MS, M / Z (ESI): 322.2 [M+H]+

[0341] Step 3: 3-{2-chloro-4'-[(1-oxo-1λ] 6 Synthesis of thiocyclopropane-1-methylene)amino][1,1'-biphenyl]-3-yl}piperidine-2,6-dione (compound 6)

[0342] 3-(3-bromo-2-chlorophenyl)piperidin-2,6-dione (A4) (100 mg, 0.33 mmol), 1-{[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl]imino}-1λ 6 Thionitropentan-1-one (6-3) (200 mg, 0.6 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (II) (31 mg, 0.04 mmol), and potassium phosphate (88 mg, 0.82 mmol) were dissolved in anhydrous N,N-dimethylformamide (1.5 mL). The reaction mixture was stirred at 100 °C for 4 h. After the reaction was complete, the reaction mixture was cooled to room temperature, filtered, and then the filtrate was purified by preparative liquid chromatography (column: Phenomenex Synergi C18 100*25 mm*4 μm; solvent: A = water + 0.1 vol% formic acid (99%), B = acetonitrile; gradient: 5%-95%, 7 min) to give compound 3-{2-chloro-4'-[(1-oxo-1λ 6 -Thiocyclopropane-1-methylene)amino][1,1'-biphenyl]-3-yl}piperidine-2,6-dione (compound 6).

[0343] 1 H NMR (400 MHz, DMSO-d6) δ 10.93 (s, 1H), 7.36-6.90 (m, 7H), 4.35-4.31 (m, 1H), 3.30-2.00 (m, 12H).

[0344] LC-MS, M / Z (ESI): 417.20 [M+H] +

[0345] Example 7: Preparation of the target compound 7

[0346] 3-[2-chloro-4'-(1,3-dimethyl-5-sulfenyl-1,5-dihydro-4H-1,2,4-triazol-4-yl)[1,1'-biphenyl]-3-yl]piperidine-2,6-dione

[0347] The synthetic route of the target compound 7 is shown as follows:

[0348] First step: synthesis of 4-(4-bromophenyl)-2,5-dimethyl-2,4-dihydro-3H-1,2,4-triazol-3-one (7-1)

[0349] Dissolve 4-(4-bromophenyl)-5-methyl-2,4-dihydro-3H-1,2,4-triazol-3-one (compound 7-1) (156.0 mg, 0.61 mmol) in N,N-dimethylformamide (8.0 mL), and add sodium hydride (49.1 mg, 1.23 mmol, 60%) under nitrogen protection at 0°C. After stirring for 10 minutes, add iodomethane (261.5 mg, 1.84 mmol), and then naturally restore to room temperature and stir overnight. After the reaction is completed, cool the reaction solution to 0°C, quench the reaction by adding water (15 mL), extract with ethyl acetate (20 mL x 3), combine the organic phases, wash with water (20 mL), and then saturated brine (20 mL), dry over anhydrous sodium sulfate, filter and concentrate, and purify the residue by silica gel column separation (petroleum ether: ethyl acetate (V / V) = 1:2) to obtain compound 4-(4-bromophenyl)-2,5-dimethyl-2,4-dihydro-3H-1,2,4-triazol-3-one (7-2).

[0350] LC-MS, M / Z (ESI): 268.1 / 270.1 [M+H] + .

[0351] Second step: synthesis of 4-(4-bromophenyl)-2,5-dimethyl-2,4-dihydro-3H-1,2,4- triazol-3-thione (7-3) 4-(4-bromophenyl)-2,5-dimethyl-2,4-dihydro-3H-1,2,4-triazol-3-one (7-2) (143.5 mg, 0.54 mmol) was dissolved in toluene (10 mL), and Lawesson's reagent (324.7 mg, 0.80 mmol) was added. The reaction was heated to 100 °C and stirred overnight. After the reaction was completed, it was cooled to room temperature, washed with water (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The reaction was purified by silica gel column separation (petroleum ether: ethyl acetate (V / V) = 8:1) to obtain compound 4-(4-bromophenyl)-2,5-dimethyl-2,4-dihydro-3H-1,2,4-triazol-3-thione (7-3) (78.5 mg, yield 51.2%).

[0352] LC-MS, M / Z (ESI): 284.1 / 286.1 [M+H] + .

[0353] Third step: synthesis of 2,5-dimethyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]- 2,4-dihydro-3H-1,2,4-triazol-3-thione (7-4) 4-(4-bromophenyl)-2,5-dimethyl-2,4-dihydro-3H-1,2,4-triazol-3-thione (7-3) (39.8 mg, 0.14 mmol) was dissolved in 1,4-dioxane (6 mL) under nitrogen protection. Pinacol diborane (55.0 mg, 0.22 mmol) and anhydrous potassium acetate (42.0 mg, 0.43 mmol) were added, followed by the addition of 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (10.5 mg, 0.014 mmol). The reaction was heated to 100 °C under nitrogen protection for 3.5 hours. After the reaction was completed, the reaction was cooled to room temperature, concentrated under reduced pressure, diluted with dichloromethane (20 mL), washed with water (10 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain crude compound 2,5-dimethyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]-2,4-dihydro-3H-1,2,4-triazol-3-thione (7-4).

[0354] LC-MS, M / Z (ESI): 332.1 [M+H]+.

[0355] Fourth step: synthesis of 3-[2-chloro-4'-(1,3-dimethyl-5-sulfenyl-1,5-dihydro-4H-1,2,4-triazol-4-yl)[1,1'- biphenyl]-3-yl]piperidine-2,6-dione (compound 7)

[0356] Dissolve 2,5-dimethyl-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]-2,4- dihydro-3H-1,2,4-triazole-3-thione (7-4) (45.7 mg, 0.14 mmol) in 1,4-dioxane (10 mL), add 3-(3-bromo-2-chlorophenyl)piperidine-2,6-dione (86 mg, 0.28 mmol) and potassium phosphate (76.0 mg, 0.36 mmol), then add 1,1'-bis(diphenylphosphino)ferrocenedichloropalladium (10.5 mg, 0.014 mmol), and stir the reaction solution at 100 °C for 12 hours under nitrogen protection. After the reaction is completed, cool the reaction solution to room temperature, concentrate under reduced pressure, and separate and purify the crude product on a silica gel column (dichloromethane:methanol (V / V) = 10:1) to obtain compound 3-[2-chloro-4'-(1,3-dimethyl-5-sulfenyl-1,5-dihydro-4H-1,2,4-triazol-4-yl)[1,1'-biphenyl]-3-yl]piperidine-2,6-dione (compound 7).

[0357] LC-MS, M / Z (ESI): 427.1 [M+H] + .

[0358] 1 H NMR (600 MHz, DMSO d6) δ 10.94 (s, 1H), 7.61 (d, 2H), 7.54 (d, 2H), 7.42 (dt, 3H), 4.37 (dd, 1H), 3.71 (s, 3H), 2.84 - 2.77 (m, 1H), 2.58 - 2.53 (m, 1H), 2.38-2.30 (m, 1H), 2.17 (s, 3H), 2.10 - 2.04 (m, 1H).

[0359] Example 8: Preparation of target compound 8

[0360] 3-[2-chloro-4'-(4-methyl-5-oxo-7-sulfanyl-4,6-diazaspiro[2.4]heptan-6-yl)[1,1'-biphenyl]-3-yl]piperidine-2,6-dione (compound 8)

[0361] The synthetic route of target compound 8 is shown as follows:

[0362] First step: synthesis of 6-(4-bromophenyl)-4-methyl-7-sulfanylidene-4,6-diazaspiro[2.4]heptan-5-one (8-1)

[0363] To a solution of 4-bromophenyl isothiocyanate (5-1) (300 mg, 1.4 mmol) in DMSO (10 mL) was added 1-(methylamino)cyclopropane-1-carboxylic acid (180 mg, 1.65 mmol), triethylamine (167 mg, 1.65 mmol) and DMAP (15 mg, 0.07 mmol) at room temperature, the reaction was heated to 65 °C and stirred for 12 h. After the reaction was completed, it was cooled to room temperature, diluted with dichloromethane (100 mL), washed with saturated aqueous sodium bicarbonate solution (50 mL*2), and the organic phase was concentrated under reduced pressure to give the crude product, which was purified by column chromatography (petroleum ether / ethyl acetate (V / V) = 1 / 0 to 1 / 1) to give compound 6-(4-bromophenyl)-4-methyl-7- thioureido-4,6-diazaspiro[2.4]heptan-5-one (compound 8-1).

[0364] LC-MS, M / Z (ESI): 311.1 [M+H] +

[0365] Second step: synthesis of 4-methyl-7-sulfanyl-6-[4-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)phenyl]-4,6-diazaspiro[2.4]heptan-5-one (8-2)

[0366] To a solution of 6-(4-bromophenyl)-4-methyl-7-thioxomethylen-4,6-diazaspiro[2.4]heptan- 5-one ((8-1) (150 mg, 0.48 mmol) in 1,4-dioxane (5 mL) was added pinacol diboronic acid (203 mg, 0.8 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (22 mg, 0.03 mmol) and potassium acetate (150.0 mg, 1.5 mmol) at room temperature, the reaction was stirred at 100 °C for 18 h under argon protection. The reaction was poured into water (10 mL), then extracted with ethyl acetate (2 mL*3), the organic phase was combined and washed with saturated brine (2 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give compound 4-methyl-7-sulfanyl-6-[4-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)phenyl]-4,6-diazaspiro[2.4]heptan-5-one (8-2) (crude), which was used directly in the next step.

[0367] LC-MS, M / Z (ESI): 359.2 [M+H] +

[0368] Step 3: Synthesis of 3-[2-chloro-4'-(4-methyl-5-oxo-7-sulfanyl-4,6-diazaspiro[2.4]heptan- 6-yl)[1,1'-biphenyl]-3-yl]piperidine-2,6-dione (Compound 8)

[0369] To a solution of 4-methyl-7-sulfanyl-6-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl]-4,6-diazaspiro[2.4]heptan-5-one crude (8-2) (150 mg, 0.48 mmol) and 3-(3- bromo-2-chlorophenyl)piperidine-2,6-dione (177 mg, 0.58 mmol) in 1,4-dioxane (5 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (22 mg, 0.03 mmol) and potassium phosphate (186 mg, 1.35 mmol), the reaction was stirred at 100 °C for 18 h under argon protection. The reaction was poured into water (10 mL), then extracted with ethyl acetate (2 mL*3). The organic phase was combined and washed with saturated brine (2 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude, which was purified by paper cup liquid chromatography (column: Phenomenex Synergi C18 100*25 mm*4 μm; solvent: A = water + 0.1 volume% formic acid (99%), B = acetonitrile; gradient: 5%-95%, 7 minutes) to give 3-[2-chloro-4'-(4-methyl-5-oxo-7-sulfanyl-4,6-diazaspiro[2.4]heptan-6-yl)[1,1'- biphenyl]-3-yl]piperidine-2,6-dione (Compound 8).

[0370] 1 H NMR (400 MHz, DMSO-d6) 10.94 (s, 1H), 7.54 (d, 2H), 7.47 - 7.36 (m, 5H), 4.37 (dd, 1H), 3.13 (s, 3H), 2.85 - 2.76 (m, 1H), 2.61 - 2.52 (m, 1H), 2.35 (dt, 1H), 2.09 - 2.03 (m, 1H), 1.96 - 1.90 (m, 2H), 1.52-1.49 (m, 2H)

[0371] LC-MS, M / Z (ESI): 454.1 [M+H] +

[0372] Example 9: Preparation of target compound 9

[0373] 3-(2-chloro-4'-{[cyclopropyl(methyl)(oxo)-lambda 6- thioalkyl]amino} [1,1'-biphenyl]-3-yl) piperidine-2,6-dione (Compound 9)

[0374] The synthetic route of the target compound 9 is shown as follows:

[0375] First step: 1-bromo-4-{[cyclopropyl(methyl)(oxo)-λ 6 Synthesis of 1-bromo-4-{[cyclopropyl(methyl)(oxo)-λ

[0376] Compound p-bromoiodobenzene (200.00 mg, 0.71 mmol) and cyclopropyl(imino)(methyl)-L6 sulfonamidone (95 mg, 0.8 mmol) were dissolved in anhydrous 1,4-dioxane solution (6 mL) at room temperature, then cesium carbonate (461 mg, 1.4 mmol), tris(dibenzylideneacetone)dipalladium (58.26 mg, 0.06 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (72.25 mg, 0.12 mmol) were added, and the reaction solution was stirred at 110°C under nitrogen protection for 12 hours. TLC monitoring showed that the raw material was completely reacted, the stirring was stopped, the reaction solution was reduced to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (10 mL x 5), the organic phase was collected, dried with anhydrous sodium sulfate, the organic phase was concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate (V / V) = 100:50) to obtain compound 1-bromo-4-{[cyclopropyl(methyl)(oxo)-λ 6 - thioalkyl]amino} [1,1'-biphenyl]-3-yl) piperidine-2,6-dione (Compound 9)

[0377] LC-MS, M / Z (ESI): 274.2 [M+H] +

[0378] Second step: 2-(4-{[cyclopropyl(methyl)(oxo)-λ 6 Synthesis of 2-(4-{[cyclopropyl(methyl)(oxo)-λ

[0379] Compound 1-bromo-4-{[cyclopropyl(methyl)(oxo)-λ 6To a solution of 2-(4-{[cyclopropyl(methyl)(oxo)-λ6-sulfanyl]amino}phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (9-2) (crude) (70 mg, 0.26 mmol) and 3-(3-bromo-2-chlorophenyl)piperidine-2,6-dione (Intermediate A4) (118 mg, 0.39 mmol) in 1,4-dioxane (5 mL) was added [1,1 '-bis(diphenylphosphino)ferrocene]dichloropalladium (22 mg, 0.03 mmol) and potassium phosphate (186 mg, 1.35 mmol) and the reaction stirred at 100 °C under nitrogen for 18 h. The reaction was poured into water (10 mL) and extracted with ethyl acetate (2 mL*3). The organic phases were combined, washed with saturated brine (2 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude. The crude was purified by preparative liquid chromatography (column: Phenomenex Synergi C18 100*25 mm*4 μm; Solvents: A = water + 0.1 % formic acid (99%), B = acetonitrile; Gradient: 5-95% over 7 minutes) to give 3-(2-chloro-4'-{[cyclopropyl(methyl)(oxo)-λ6-sulfanyl]amino}[1,1 '-biphenyl]-3-yl)piperidine-2,6-dione (Compound 9). 6 2-(4-{[cyclopropyl(methyl)(oxo)-λ6-sulfanyl]amino}phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (9-2) (crude) was used directly in the next step.

[0380] LC-MS, M / Z (ESI): 322.2 [M+H] +

[0381] Third step: 3-(2-chloro-4'-{[cyclopropyl(methyl)(oxo)-λ6-sulfanyl]amino}[1,1 '-biphenyl]-3-yl)piperidine-2,6-dione (Compound 9) 6 2-(4-{[cyclopropyl(methyl)(oxo)-λ6-sulfanyl]amino}phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (9-2) (crude) was used directly in the next step.

[0382] To a solution of 2-(4-{[cyclopropyl(methyl)(oxo)-λ6-sulfanyl]amino}phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (9-2) (crude) (70 mg, 0.26 mmol) and 3-(3-bromo-2-chlorophenyl)piperidine-2,6-dione (Intermediate A4) (118 mg, 0.39 mmol) in 1,4-dioxane (5 mL) was added [1,1 '-bis(diphenylphosphino)ferrocene]dichloropalladium (22 mg, 0.03 mmol) and potassium phosphate (186 mg, 1.35 mmol) and the reaction stirred at 100 °C under nitrogen for 18 h. The reaction was poured into water (10 mL) and extracted with ethyl acetate (2 mL*3). The organic phases were combined, washed with saturated brine (2 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude. The crude was purified by preparative liquid chromatography (column: Phenomenex Synergi C18 100*25 mm*4 μm; Solvents: A = water + 0.1 % formic acid (99%), B = acetonitrile; Gradient: 5-95% over 7 minutes) to give 3-(2-chloro-4'-{[cyclopropyl(methyl)(oxo)-λ6-sulfanyl]amino}[1,1 '-biphenyl]-3-yl)piperidine-2,6-dione (Compound 9). 6 3-(2-chloro-4'-{[cyclopropyl(methyl)(oxo)-λ6-sulfanyl]amino}[1,1 '-biphenyl]-3-yl)piperidine-2,6-dione (Compound 9).

[0383] 1H NMR (400 MHz, DMSO-d6) δ 10.90 (s, 1H), 7.39-7.32 (m, 2H), 7.27 (dd, 2H), 7.21 (t, 2H), 6.98 (d, 1H), 4.31 (dd, 1H), 3.20 (s, 3H), 2.85-2.81 (m, 1H), 2.79 - 2.73 (m, 1H), 2.55 - 2.50 (m, 1H), 2.32 - 2.27 (m, 1H), 2.05 - 2.00 (m, 1H), 1.18 (dt, 1H), 1.13 - 1.08 (m, 1H), 1.07 - 0.98 (m, 2H).

[0384] LC-MS, M / Z (ESI): 417.1 [M+H] +

[0385] Example 10: Preparation of compound 10:

[0386] 3-(2-chloro-4'-{[4-(cyclopropanecarbonyl)-1-oxo-1λ 6 - thiamethane-1 -methylidene] amino} [1, 1 '-biphenyl]-3-yl)piperidine-2,6-dione (compound 10)

[0387] Reference Example 6: Preparation of compound 3-{2-chloro-4'-[(1-oxo-1λ 6 - thiamethane-1 -methylidene) amino] [1, 1 '-biphenyl]-3-yl}piperidine-2,6-dione (compound 6) was synthesized according to the procedure described in Reference Example 5 to give 3-(2-chloro-4'-{[4-(cyclopropanecarbonyl)-1-oxo-1λ 6 - thiamethane-1 -methylidene] amino} [1, 1 '-biphenyl]-3-yl)piperidine-2,6-dione (compound 10); LC-MS, M / Z (ESI): 500.1 [M+H] + .

[0388] Example 11 : Preparation of compound 11 :

[0389] 3-[2-chloro-4'-(3,4,4-trimethyl-5-oxo-2-sulfanylimidazolidin-1-yl)[1,1'-biphenyl]-3- yl]piperidine-2,6-dione (compound 11)

[0390] 3-[2-chloro-4'-(3,4,4-trimethyl-5-oxo-2-sulfanylimidazolidin-1-yl)[1,1'-biphenyl]-3- yl]piperidine-2,6-dione (Compound 11) was synthesized according to the procedure described in Reference Example 8 for the preparation of 3-[2-chloro-4'-(4-methyl-5-oxo-7- sulfanyl-4,6-diazaspiro[2.4]heptan-6-yl)[1,1'-biphenyl]-3-yl]piperidine-2,6-dione (Compound 8); LC-MS, M / Z (ESI): 456.1 [M+H] + .

[0391] The following compounds were synthesized according to the procedure described in Reference Example 1:

[0392] Biological test

[0393] Test Example 1: Compound-induced binding of VAV1 to CRBN

[0394] Experimental method

[0395] 1) Cell line construction and culture. VAV1-SmBiT expression plasmid and CRBN-LgBiT expression plasmid were constructed, respectively, and VAV1-SmBiT and CRBN-LgBiT were inserted into the genome of HEK293 cells using a lentivirus system to construct a cell line (HEK293-VAV1-SmBiT-CRBN-LgBiT) stably expressing VAV1-SmBiT and CRBN-LgBiT proteins. The culture medium used for culture was DMEM medium containing inactivated 10% fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin, and the HEK293-VAV1-SmBiT-CRBN-LgBiT cells were cultured in a 37°C, 5% CO2 incubator. When the cell confluence reached 80-90%, the cells were subcultured. Cells in the logarithmic growth phase were used for plating, and HEK293-VAV1-SmBiT-CRBN-LgBiT cells were plated in a 96-well plate at 100 μL of culture medium per well, 20,000-30,000 cells per well, and incubated overnight.

[0396] 2) Compound dilution. The compound was dissolved in DMSO to a concentration of 10 mM. Subsequently, the compound was gradient-diluted with DMSO to concentrations of 2000 μM, 500 μM, 125 μM, 32.25 μM, 7.81 μM, 1.95 μM, 0.49 μM, 0.12 μM, and 0.031 μM, respectively.

[0397] 3) Drug addition. Take 1 μL of the diluted compound and add it to 1 mL of complete medium, mix well, and then take 100 μL and add it to a 96-well plate, so that the working concentration is 1000 nM, 250 nM, 62.5 nM, 15.6 nM, 3.91 nM, 0.98 nM, 0.24 nM, 0.061 nM, 0.015 nM, and 0 nM.

[0398] 4) Activity detection. Detection is performed after 8 hours of drug treatment, and the detection method is according to the NanoBiT Protein: Protein Interaction System instruction book of promega company.

[0399] 5) EC 50 Calculation. Fluorescence intensity (Fold change) = (Lumninence experiment - Lumninence blank) / (Lumninence DMSO - Lumninence blank).

[0400] Experimental conclusion: The compound of the present application can induce the direct binding of VAV1 and CRBN in a dose-dependent manner.

[0401] Test example 2: Effect of the compound on VAV1 protein of HEK293 cells

[0402] Experimental method

[0403] 1) Cell line construction and culture. The C-terminal of VAV1 is inserted into a Hibit tag, and a lentiviral system is used to insert VAV1-Hibit into the genome of HEK293 cells to construct a cell line stably expressing VAV1-Hibit protein. The culture medium used for culture is DMEM medium containing inactivated 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin, and the HEK293-VAV1-Hibit cells are cultured in a 37°C, 5% CO2 incubator. The cell confluence rate reaches 80-90% after subculture. Cells in the logarithmic growth phase are used for plating, and HEK293-VAV1-Hibit cells are plated in a 96-well plate, 100 μL of medium per well, 20,000-30,000 cells per well, and incubated overnight.

[0404] 2) Compound dilution. The compound is dissolved in DMSO to a concentration of 10 mM. Then the compound is gradient-diluted with DMSO to concentrations of 2000 μM, 500 μM, 125 μM, 32.25 μM, 7.81 μM, 1.95 μM, 0.49 μM, 0.12 μM, and 0.031 μM.

[0405] 3) Drug addition. Take 1 μL of the diluted compound and add it to 1 mL of complete medium, mix well, and then take 100 μL and add it to a 96-well plate, so that the working concentration is 1000 nM, 250 nM, 62.5 nM, 15.6 nM, 3.91 nM, 0.98 nM, 0.24 nM, 0.061 nM, 0.015 nM, and 0 nM.

[0406] 6) Activity detection. Detection is performed after 24 hours of drug treatment, and the detection method is according to the instructions of the HiBiT Lytic Detection System of Promega.

[0407] 7) DC 50 Calculation. Relative VAV1 (%) = (Lumninence experiment - Lumninence blank) / (Lumninence DMSO - Lumninence blank), calculated according to the fitting of log(inhibitor) vs. response - Variable slope (four parameters).

[0408] Experimental conclusion: The compound of the present application shows excellent degradation effect on VAV1 protein and is dose-dependent.

[0409] Test Example 3: Degradation of VAV1 of Jurkat cells by the compound

[0410] Jurkat cells are cultured in RPMI-1640 medium containing inactivated 10% fetal bovine serum, 100 U / ml penicillin and 100 μg / ml streptomycin in a 37℃, 5% CO2 incubator, and the cell density reaches 1×10^6, then subcultured and divided into bottles. Tumor cells in the logarithmic growth phase are plated in a 12-well plate at a density of 7×10 5 The drug is diluted and added to the cell culture medium, so that the final concentration of the drug is 1000 nM, 250 nM, 62.5 nM, 15.6 nM, 3.9 nM, 0.98 nM, 0.24 nM, and 0 nM. The cells are treated with the drug for 24 hours. The cells treated with the drug for 24 hours are taken out of the incubator and transferred to a 1.5 mL EP tube, and the supernatant is removed by centrifugation. Add 150 μL of RIPA lysis buffer to each tube and incubate on ice for 30 minutes. Run the protein gel for detection. Anti-VAV1 is diluted 1:1000, Anti-GAPDH is diluted 1:5000, and HRP secondary antibody is diluted 1:10000, and incubate at 4℃ overnight. RT for 1 hour. Detect protein expression by chemiluminescence and analyze gray value by ImageJ. The experimental results are shown in Table 1.

[0411] ​Table 1 Compound degradation activity of VAV1 protein in Jurkat cells.

[0412] *DC50≤10nM is“A”;10nM<DC50≤30nM is“B1”, 30nM<DC50≤100nM is“B2”;100nM<DC50≤1000nM is“C”;DC50>1000nM is“D”.

[0413] Experimental conclusion: The compound of the present application shows excellent degradation effect on the degradation of VAV1 protein in Jurkat cells, and shows dose-dependent.

[0414] Test example 4: Compound inhibits CD3 / CD28-induced Jurkat cell activation

[0415] Jurkat cells were cultured in RPMI-1640 medium containing inactivated 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin at 37℃ in a 5% CO2 incubator, and the cell density reached 1×10^6, then passaged and divided into bottles. Cells in logarithmic growth phase were plated at 40000 in a 96-well plate, and different concentrations of compounds were added. After 24 hours of treatment, 5 μg / mL CD3 antibody and 1 μg / mL CD28 antibody were added, and after 24 hours of continuous culture. Take the cell supernatant and detect the IL-2 content by ELISA.

[0416] Experimental conclusion: In the CD3 / CD28-induced Jurkat cell activation model, the compound of the present application can significantly inhibit the production of IL-2, and the inhibition function is positively correlated with the dose.

[0417] Test example 5: Compound inhibits CD3 / CD28-induced human T cell activation

[0418] Fresh blood sample was diluted with equal volume of PBS, then slowly added into 50 mL centrifuge tube containing 15 mL Lymphoprep (Stemcell, #7851), without breaking the interface, centrifuged at 1000 g, 5 min, 25 min, without brake. The white blood cell layer (PBMC) was collected, washed twice with PBS, centrifuged at 350 g for 10 min, and the supernatant was discarded. T cells were sorted from PBMC using T cell sorting kit (Stemcell, #17951), and the cell density was adjusted to 2.6 x 106 / mL, 75 μL was added to each well of a U-bottom 96-well plate (Corning, #3799), and 75 μL of the diluted test compound was added, and the blank group was added with an equal amount of DMSO in 1640 complete medium, mixed and incubated for 24 hours. Then the cells were transferred to a 96-well plate coated with Anti-Human CD3 (5 μg / mL, BD, #555329), and 50 μL of Anti-Human CD28 (1 μg / mL, BD, #555725) was added, mixed and incubated for another 48 hours. Finally, the supernatant was collected, and the IL-2 level was detected using an IL-2 ELISA kit (BD, #555190).

[0419] Experimental conclusion: In the CD3 / CD28-induced human T cell activation model, the compound of the present application can significantly inhibit the production of IL-2, and the inhibition function is positively correlated with the dose.

[0420] Test Example 6: Mouse pharmacokinetic test

[0421] The mouse pharmacokinetic test used male ICR mice, 20-25 g, and the mice were fasted overnight. Three mice were taken, and 10 mg / kg was administered orally by gavage. Blood was collected before administration and at 15, 30 minutes, and 1, 2, 4, 8, and 24 hours after administration. The blood sample was centrifuged at 6800 g for 6 minutes at 2-8°C, and the plasma was collected and stored at -80°C. The plasma at each time point was mixed with 3-5 times the amount of internal standard acetonitrile solution, vortexed for 1 minute, centrifuged at 13000 rpm for 10 minutes at 4°C, and the supernatant was mixed with 3 times the amount of water. An appropriate amount of the mixture was taken for LC-MS / MS analysis. The main pharmacokinetic parameters were analyzed using WinNonlin 7.0 software non-compartment model.

[0422] Table 2: Compound mouse PK parameters

[0423] The results of the mouse pharmacokinetic test showed that the compound of the present application exhibited excellent pharmacokinetic properties and good drug development potential.

[0424] Test Example 7: Rat pharmacokinetic test

[0425] Rat pharmacokinetic test, using male SD rats, 180-240g, fasting overnight. Take 3 rats, oral gavage administration of 10mg / kg. Before administration and after administration of 15, 30 minutes and 1, 2, 4, 8, 24 hours, blood sampling. Blood samples 6800g, 2-8℃ centrifugation for 6 minutes, collect plasma, stored at -80℃. Take the plasma at each time point, add 3-5 times the amount of internal standard acetonitrile solution, mix, vortex mix for 1 minute, 13000r / min, 4℃ centrifugation for 10 minutes, take the supernatant, add 3 times the amount of water, take the appropriate amount of mixture for LC-MS / MS analysis. The main pharmacokinetic parameters are analyzed by WinNonlin 7.0 software non-compartment model.

[0426] The results of the rat pharmacokinetic test show that the compound of the application has excellent rat pharmacokinetic properties and good drug development potential.

[0427] Test Example 8: Human liver microsomal stability test

[0428] The human liver microsomal stability test is detected by co-incubation of the compound with human liver microsomes in vitro. First, the test compound is prepared into a 10mM stock solution in DMSO solvent, then the compound is diluted to 0.5mM using acetonitrile. Dilute human liver microsomes (Corning) into a microsomal / buffer solution using PBS, and dilute the 0.5mM compound into a working solution using the solution, the compound concentration in the working solution is 1.5μM, and the human liver microsomal concentration is 0.75mg / mL. Take a deep well plate, add 30μL of the working solution to each well, then add 15μL of preheated 6mM NADPH solution to start the reaction, and incubate at 37℃. At 0, 5, 15, 30, and 45 minutes of incubation, add 135μL of acetonitrile to the corresponding wells to terminate the reaction. After the last 45-minute time point, vortex the deep well plate for 10 minutes (600rpm / min) and then centrifuge for 15 minutes. After centrifugation, take the supernatant, add purified water 1:1, and then perform LC-MS / MS detection. Obtain the peak area ratio of the compound to the internal standard at each time point, compare the peak area ratios of the compound at 5, 15, 30, and 45 minutes with the peak area ratio at 0 minutes, calculate the remaining percentage of the compound at each time point, and calculate T1 / 2 using Graphpad 5 software.

[0429] The results of the human liver microsomal stability test show that the compound of the application has excellent human liver microsomal stability and good drug development potential.

[0430] Test Example 9: Thermodynamic solubility test

[0431] The solubility of the compounds was evaluated by standardized in vitro methods. The control compounds progesterone and diclofenac were purchased from Sigma. PBS (pH 7.4), FaSSIF, FeSSIF and FaSSGF buffers were freshly prepared in the laboratory according to standard recipes and used within the specified time. The solubility determination procedure was as follows: about 1 mg of compound was weighed into a glass bottle, the corresponding buffer (1 mL / mg) was added, a stirring bar was added and sealed, and the bottle was shaken at 25°C, 1100 rpm for 24 hours. After incubation, the stirring bar was removed, and the sample was filtered under vacuum and diluted in gradient of 100, 1000, 10000 times, and the diluent was a mixture of water / acetonitrile (1:1) containing an internal standard. The standard was dissolved in DMSO and diluted in the same way to concentrations of 10, 1, 0.1 μg / mL. All samples were placed in an autosampler and analyzed using LC-MS / MS. Data processing was performed using Excel software, and the solubility was calculated by comparison with the standard.

[0432] The results of the thermodynamic solubility test show that the compounds of the present application have good thermodynamic solubility and good drug properties.

[0433] Test Example 10: Inhibition test of compounds on cytochrome P450

[0434] The inhibition potential of the compounds on cytochrome P450 (CYP450) subtype CYP3A4 (2 substrates, midazolam and testosterone) was detected. The test compound was first prepared into a 10 mM stock solution in DMSO solvent, and the CYP3A4 inhibitor ketoconazole was prepared into a 10 mM, 2.5 mM, 2.5 mM stock solution in DMSO solvent. The test compound and ketoconazole were diluted with acetonitrile to a final concentration of 400 times (compound: 10 μM, ketoconazole: 2.5 μM).

[0435] The NADPH cofactor (66.7 mg NADPH in 10 mL potassium phosphate buffer) and the substrates were prepared in a 4-fold final concentration of potassium phosphate buffer (0.1 M, pH 7.4). The final concentration of the CYP3A4 substrate midazolam was 320 μM, and the final concentration of the CYP3A4 substrate testosterone was 20 μM.

[0436] Human liver microsomes solution was prepared in potassium phosphate buffer on ice at a concentration of 0.2 mg / mL. The test compound and control inhibitor solutions were prepared in human liver microsomes solution on ice at a 2x final concentration. 30 μL of test compound and control inhibitor solutions were added to the test wells, and 15 μL of substrate was added in duplicate. The 96-well assay plate and NADPH solution were incubated at 37 °C for 5 minutes, and 15 μL of pre-warmed 8 mM NADPH solution was added to the assay plate to initiate the reaction. The CYP3A4 assay plate was pre-incubated at 37 °C for 5 minutes. The reaction was terminated by adding 120 μL of acetonitrile, and after quenching, the plate was shaken on a shaker (IKA, MTS2 / 4) for 10 minutes at 600 rpm / min, and then centrifuged for 15 minutes. After centrifugation, the supernatant was taken, and 1:1 purified water was added, and then LC-MS / MS detection was performed to obtain the ratio of compound peak area to internal standard peak area. The peak area ratio of the compound was compared with the peak area ratio of the control inhibitor, and the inhibition rate was calculated.

[0437] The results of the cytochrome P450 inhibition test show that the compound of the present application has no obvious CYP3A4 (two substrates, midazolam and testosterone) inhibition effect, and has good drug property.

[0438] Test Example 11: Test of plasma protein binding by equilibrium dialysis method

[0439] First, the human or other species' plasma samples were stored in a -20℃ refrigerator, thawed in a 37℃ water bath before use, and kept on wet ice for standby. The working solution of the test compound was prepared using DMSO, with a stock concentration of 10 mM and a final concentration of 2 μM. The thawed plasma was centrifuged to remove suspended impurities and precipitates, and the pH was adjusted to the range of 7.0-8.0. According to the manufacturer's instructions, the pretreated dialysis membrane was installed in the dialysis device and the installation was completed. When preparing the zero-time control sample, the blank plasma was mixed with the working solution of the test compound, and a vortex mixer was used to mix at 1000 rpm for 2 minutes to obtain a final concentration of 2 μM, which was immediately transferred to a 96-well plate as the T=0 time point control sample, and the remaining mixture was placed in a constant temperature incubator for continuous incubation. To determine the stability of the compound in the plasma, the remaining mixture was incubated in a 37℃ constant temperature shaking incubator for 5 hours, and 50 μL of the sample was transferred to a 96-well plate for subsequent analysis after incubation. The dialysis operation was balanced according to the manufacturer's instructions to assemble the dialysis device, the plasma sample was added to the dialysis cavity, and dialysis was performed with an equal volume of PBS buffer. The experiment was repeated, the device was covered with an air-permeable cover, and incubation was performed at 37℃ and 100 rpm for 5 hours. After incubation, samples were taken from the plasma cavity and the buffer cavity and transferred to a 96-well plate for analysis. During sample processing, 500 μL of 80% acetonitrile / methanol solution containing appropriate internal standards was added to the collected samples and mixed for 2 minutes. The protein was precipitated and the compound was released by adding 500 μL of 80% acetonitrile / methanol solution containing appropriate internal standards, vortexing for 10 minutes, and centrifuging at 4000 rpm for 10 minutes. 100 μL of supernatant was transferred to a new 96-well plate, mixed with 300 μL of distilled water, and then subjected to LC-MS / MS analysis. All samples were automatically peak area integrated, and the analyte peak area and internal standard peak area were exported to an Excel table. The free rate, binding rate and recovery rate of the compound were calculated according to the following formula: Free rate (% Unbound) = (buffer cavity peak area ratio / plasma cavity peak area ratio) x 100; Binding rate (% Bound) = 100-% Unbound; Recovery rate (% Recovery) = (buffer cavity peak area ratio + plasma cavity peak area ratio) / total sample peak area ratio x 100; Remaining amount (% Remaining) = 5-hour peak area ratio / 0-hour peak area ratio x 100.

[0440] The experimental results show that the compound of the present application has a significant advantage in plasma protein binding rate.

[0441] Test Example 12: Evaluation of the pharmacodynamic effect of the compound in a mouse inflammatory bowel disease model induced by adoptive T cell transfer

[0442] This example aims to evaluate the therapeutic potential of the compound in a mouse inflammatory bowel disease model. The model used is an inflammatory bowel disease model induced by adoptive T cell transfer, which has good clinical relevance.

[0443] Experimental Methods:

[0444] 1) Isolate spleen from BALB / c donor mice, prepare single cell suspension, sort CD4 + CD25 - cells by EasySep kit, sort CD4 + CD45RB + cells by flow cytometry, wash with PBS and resuspend to 1.5 x 10 6 / mL. Then transfer to immunodeficient recipient mice (CB17-SCID mice) by tail vein injection.

[0445] 2) Randomly group the recipient mice on day 15 after cell transfer, and give them compounds or control solvent respectively. Test compounds are administered by oral gavage, at a dose range of 0.1-10 mg / kg, once daily for 20 days.

[0446] 3) During the experiment, record the changes in body weight, stool character and activity of the mice. After cell inoculation, score DAI twice a week for the first and second weeks, and score DAI three times a week for the third to fifth weeks. The DAI score is the sum of the body weight loss and stool consistency scores, through the changes in animal body weight and stool consistency.

[0447] 4) Sacrifice the mice on day 35, and collect the colon tissues. At the endpoint, collect the colon, measure the length, wash with PBS and remove the feces, weigh after blotting, and calculate the weight to length ratio. At the endpoint dissection, clean the colon contents, and save for pathological scoring.

[0448] The results show that the compound group of the present application significantly alleviates the disease progression of the mouse intestinal inflammation model induced by adoptive naive T cells.

Claims

1. A compound represented by Formula (II), a tautomer, a stereoisomer, a pharmaceutically acceptable salt, or a prodrug thereof, ###00001### (II) wherein, R1is halogen, OCF3, CN, C 2-6 alkyl, -OC 2-6 alkyl, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, 4-10 membered heterocycloalkyl, said C 2-6 alkyl, -OC 2-6 alkyl, C 3-6 cycloalkyl, C 2-6 alkynyl and 4-10 membered heterocycloalkyl are each independently optionally substituted with 1, 2, 3, or 4 R 1a substituents; Each R 1a They are, independently, H, halogen, OH, NH2, CN, and C. 1-6 Alkyl or halogenated C 1-6 alkyl; R2, R3and R4are each independently H, halogen, NH2, CN, C 1-6 alkyl or haloC 1-6 alkyl; X1is N or CR5; X2is N or CR 51 ; X3is N or CR7; X4is N or CR 71 ; R5, R 51 , R7and R 71 are each independently H, halogen, NH2, CN or C 1-6 alkyl or halogen C 1-6 alkyl; R8is H, D, halogen or C 1-6 alkyl; L1is a single bond, -O-, -NH-, -O-C 1-3 alkyl-, -NH-C 1-3 alkyl-, -C 1-3 alkyl- or -C 3-6 cycloalkyl-, wherein said -NH-, -O-C 1- 3alkyl-, -NH-C 1-3 alkyl-, -C 1-3 alkyl- and -C 3-6 cycloalkyl- are each independently optionally substituted with 1, 2, 3 or 4 R 1L substituents; each R is independently H, halogen, C 1L each R is independently H, halogen, C 1-3 alkyl or C 3-6 cycloalkyl; R6 represents CN, -C 0-3 Alkyl-P(=O)(R) 61 )2、-C 0-3 Alkyl-(N=)S(=O)(R 61 )2、-C 0-3 Alkyl-S(=O)(=NR) 62 )R 63 C 1-6 Alkyl, 4-10-membered heterocyclic alkyl, 5-10-membered heterocyclic alkenyl, 6-10-membered aryl or 5-10-membered heteroaryl, wherein C 1-6 Alkyl, 4-10-membered heterocyclic alkyl, 5-10-membered heterocyclic alkenyl, 6-10-membered aryl, and 5-10-membered heteroaryl groups are each independently and optionally bounded by 1, 2, 3, 4, 5, 6, 7, or 8 R groups. 6a replace; R 61 , R 62 , and R 63 are each independently H, C 1-6 alkyl or C 3-6 cycloalkyl; Each R 6a The independent components are H, halogen, oxo (=O), thio (=S), OH, NH2, CN, and C, respectively. 1-6 Alkyl, Halogenated C 1-6 Alkyl or C 3-6 cycloalkyl; or two R 61 and the P or S atom to which they are attached together form a 4-10 membered heterocycloalkyl group, which is optionally substituted by 1, 2, 3 or 4 R 6b substituents; Or, R 62 With R 63 Together with the N and S atoms attached thereto, they form a 5-10 membered heterocyclic alkenyl group, which is optionally bounded by 1, 2, 3 or 4 R atoms. 6b replace; each R is independently H, halogen, oxo (=0), OH, NH2, CN, C 6b each R is independently H, halogen, oxo (=0), OH, NH2, CN, C 1-6 alkyl, haloC 1-6 alkyl, C 3-6 cycloalkyl, -L2-C 1-6 alkyl, -L2-C 3-6 cycloalkyl or -L2-4-8 membered heterocycloalkyl, each of which is independently optionally substituted with 1, 2, 3, or 4 R 1-6 alkyl, haloC 1-6 alkyl, C 3-6 cycloalkyl, -L2-C 1-6 alkyl, -L2-C 3-6 cycloalkyl or -L2-4-8 membered heterocycloalkyl, each of which is independently optionally substituted with 1, 2, 3, or 4 R each L2is independently -O-, -N(R 2L )-, -S-, -S(=O)-, -S(=O)2-, or -C(=O)-; R 2L is H or C 1-3 alkyl; each R is independently H, halogen, OH, NH2, or CN; with the proviso that when R1is halogen, R6is -C 0-3 alkyl P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, -C 0-3 alkyl-S(=O)(=NR 62 )R 63 , 4-10 membered heterocycloalkyl or 5-10 membered heterocycloalkenyl, said 4-10 membered heterocycloalkyl and 5-10 membered heterocycloalkenyl being substituted with at least one thioxo (C=S); the heteroatom groups in the "heterocycloalkyl", "heterocycloalkenyl", and "heteroaryl" comprise N, O, S, S(=O), S(=O)2, or S(=O)(=NH), the number of the heteroatom groups is 1, 2, 3, or 4; when the number of the heteroatom groups is plural, the heteroatom groups are the same or different.

2. The compound, tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug thereof of claim 1, wherein, which satisfies one or more of the following conditions: (1) R1is Cl, CN, -OCF3, C 2-4 alkyl, C 3-4 alkyl, C 2-4 alkyl, C 3-4 alkyl, C 1a substituted; (2) each R is independently H or F; 1a are each independently H or F; (3) R6is 4-8 membered heterocycloalkyl, 5-8 membered heterocycloalkenyl, or 5-6 membered heteroaryl, each independently optionally substituted with 1, 2, 3, 4, or 5 R 6a substituents; (4) Each R 6a They can be independently H, oxo (=O), thio (=S), CH3, -CH2CH3, CHF2, CF3 or cyclopropyl; (5) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, or -C 0-3 alkyl-S(=O)(=NR 62 )R 63 ; (6) R 61 , R 62 , and R 63 are each independently H, CH3, -CH2CH3, or cyclopropyl; (7) two R 61 and the P or S atom to which they are attached together form a 4-6 membered heterocycloalkyl group, optionally substituted by 1, 2, 3 or 4 R 6b groups; (8)R 62 With R 63 Together with the N and S atoms attached thereto, they form a 5-10 membered heterocyclic alkenyl group, which is optionally bounded by 1, 2, 3 or 4 R atoms. 6b replace; (9) each R 6b are each independently H, halogen, oxo (=0), OH, NH2, CN, C 1-3 alkyl, haloC 1-6 alkyl, C 3-6 cycloalkyl, -L2-C 1-3 alkyl, -L2-C 3-5 cycloalkyl or -L2- 4-6 membered heterocycloalkyl, said C 1-3 alkyl, haloC 1-6 alkyl, C 3-6 cycloalkyl, -L2-C 1-3 alkyl, -L2-C 3-5 cycloalkyl or -L2- 4-6 membered heterocycloalkyl are each independently optionally substituted with 1, 2, 3, or 4 R; (10) each L2is independently -O-, -N(R 2L )- or -C(=O)-; (11) R 2L is H; (12) R6is 4-8 membered heterocycloalkyl or 5-8 membered heterocycloalkenyl, said 4-8 membered heterocycloalkyl and 5-8 membered heterocycloalkenyl being substituted with 1, 2, 3, 4, 5 or 6 R 6a substituents, wherein at least one R 6a is thio (=S); (13) L1is selected from the group consisting of a single bond, -C 1-3 alkyl-, -O-C 1-3 alkyl-, -C 3-6 cycloalkyl-.

3. The compound, tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug thereof of claim 1, wherein which satisfies one or more of the following conditions: (1a) R1is Cl, CN, -OCF3, -C≡C-CH3, (2a) R2, R3, and R4are each independently H; (3a) R5, R 51 , R7and R 71 are each independently H; (4a) L1is a single bond, -CH2-, -O-CH2-, or (5a) R6is (6a) R6is (7a) each R 6b are each independently H, -C(=O)-C 1-3 alkyl, or -C(=O)-C 3-5 cycloalkyl; (8a) each R 6b are each independently H, -C(=0)-CH3, -C(=0)-CH2CH3, or -C(=0)-cyclopropyl; (9a) when R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, 4-10 membered heterocycloalkyl, or 5-10 membered heterocycloalkenyl, wherein two R 61 and the P or S atom to which they are attached together form a heterocycloalkyl group, R6is (10a) R6 is (11a) R8is H or D; (12a) R6is -C 0-3 alkyl-S(=0)(=NR 62 )R 63 wherein R 62 and R 63 together with the N and S atoms to which they are attached form a 5-10 membered heterocycloalkenyl group, R6is 4. The compound, tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug thereof of claim 1, wherein, which satisfies one or more of the following conditions: (1 b) when R1is CI, R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, or -C 0-3 alkyl-S(=O)(=NR 62 )R 63 ; (2b) when R1is Cl, R6is 4-8 membered heterocycloalkyl or 5-8 membered heterocycloalkenyl, the 4-8 membered heterocycloalkyl and 5-8 membered heterocycloalkenyl being substituted with at least one thioxo (=S); (3b) R6is (4b) when R1is Cl, R6is 5. The compound, tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug thereof of claim 1, wherein, The compound is selected from the following structures: the definitions of R1, R8are as described in claim 1, wherein L1and R6satisfy one or more of the following conditions: (1c) L1is a single bond, -CH2-, -0-CH2-, or (2c) R6is 4-8 membered heterocycloalkyl, 5-8 membered heterocycloalkenyl, or 5-6 membered heteroaryl, each independently optionally substituted with 1, 2, 3, 4, or 5 R 6a substituents; each R 6a is independently H, oxo (=0), thioxo (=S), CH3, -CH2CH3, CHF2, CF3, or cyclopropyl. (3c) R6is (4c) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, or -C 0-3 alkyl-S(=O)(=NR 62 )R 63 ; each of R 61 , R 62 , and R 63 is independently H, CH3, -CH2CH3, or cyclopropyl; (5c) R6is (6c) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, two R 61 groups together with the P or S atom to which they are attached form a heterocycloalkyl group, said heterocycloalkyl group being a 4-6 membered heterocycloalkyl group, said 4-6 membered heterocycloalkyl group being optionally substituted with 1, 2, 3, or 4 R 6b groups; each R 6b is independently H, halogen, oxo (=O), OH, NH2, CN, C 1-3 alkyl, -L2-C 1-3 alkyl, -L2-C 3-5 cycloalkyl, or -L2-4-6 membered heterocycloalkyl; wherein L2is -O-, -NH-, or -C(=O)-; (7c) R6is The R 6b Hydrogen, L2-C 1-3 Alkyl, L2-C 3-5 Cycloalkyl, L2-4-6-membered heterocycloalkyl, wherein L2 is -C(=O)-; (8c) R6is R1is hydrogen, 6b hydrogen, (9c)R6 is -C 0-3 Alkyl-S(=O)(=NR) 62 )R 63 The R 62 With R 63 Together with the N and S atoms attached thereto, they form a 5-10 membered heterocyclic alkenyl group, wherein the 5-10 membered heterocyclic alkyl group is optionally surrounded by 1, 2, 3 or 4 R atoms. 6b When replacing, R6 is (10c) R6is 6. The compound, tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug thereof of claim 1, wherein The compound is selected from the following structures: the definitions of L1and R8are as described in claim 1, wherein R6satisfy one or more of the following conditions: (1d) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, or -C 0-3 alkyl-S(=O)(=NR 62 )R 63 ; (2d) R6is (3d) R6is 4-8 membered heterocycloalkyl or 5-6 membered heterocycloalkenyl, the 4-8 membered heterocycloalkyl and 5-6 membered heterocycloalkenyl being substituted with at least one thioxo (=S); (4d) R6is 7. The compound, tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug thereof of claim 1, wherein which satisfies one or more of the following conditions: (1e) R6is 4-8 membered heterocycloalkyl, 5-8 membered heterocycloalkenyl, or 5-6 membered heteroaryl, each of which is independently optionally substituted with 1, 2, 3, 4, or 5 R 6a substituents; or, (2e) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, or -C 0-3 alkyl-S(=O)(=NR 62 )R 63 , wherein each R 61 , R 62 and R 63 is independently H, C 1-6 1-6 alkyl, or C 3-6 3-6 cycloalkyl; or, (3e) R6is -C 0-3 alkyl-P(=O)(R 61 )2, -C 0-3 alkyl-(N=)S(=O)(R 61 )2, both R 61 groups and the P or S atom to which they are attached together form a 4-6 membered heterocycloalkyl group, which is optionally substituted with 1, 2, 3, or 4 R 6b groups; or, (4e)R6 is -C 0-3 Alkyl-S(=O)(=NR) 62 )R 63 The R 62 With R 63 Together with the N and S atoms attached thereto, they form a 5-10 membered heterocyclic alkenyl group, which is optionally bounded by 1, 2, 3 or 4 R atoms. 6b Replace; or, (5e) R6is 4-8 membered heterocycloalkyl or 5-8 membered heterocycloalkenyl, which 4-8 membered heterocycloalkyl and 5-8 membered heterocycloalkenyl is substituted with 1, 2, 3, 4, 5 or 6 R 6a substituents, wherein at least one R 6a is thio (= S).

8. The compound, tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug thereof of claim 1, wherein, which satisfies one or more of the following conditions: (1f) R6is Preferably, or, (2f) R6is or, (3f) R6is Preferably, R6 is or, (4f) R6is or (5f) R6is Preferably, R6 is 9. The compound, tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug thereof of claim 1, wherein, R6is selected from -(N=)S(=0)(R 61 )2, 4-7 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl; said R 61 is each independently CH3, CHF2, or cyclopropyl; said 4-7 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl is optionally substituted with 1, 2, 3, 4, or 5 R 6a , wherein at least one R 6a is selected from thio (C=S); or, two R 61 , together with the S atom to which they are attached, form a 5-6 membered heterocycloalkyl, said 5-6 membered heterocycloalkyl is optionally substituted with 1, 2, 3, or 4 R 6b ; R6is selected from Preferably, R6is selected from More preferably, R6is selected from 10. The compound, tautomer, stereoisomer, pharmaceutically acceptable salt, or prodrug thereof of claim 1, wherein, The compound has the structure:

11. A pharmaceutical composition, characterized by, a therapeutically effective dose of the compound according to any one of claims 1-10, a tautomer, a stereoisomer, a pharmaceutically acceptable salt, or a prodrug thereof, and a pharmaceutically acceptable excipient.

12. Use of the compound according to any one of claims 1-10, a tautomer, a stereoisomer, a pharmaceutically acceptable salt, or a prodrug thereof, or the pharmaceutical composition according to claim 11 in the manufacture of a medicament for treating or preventing a VAV1 -related disease.

13. Use according to claim 12, characterized in that, The VAV1 -related disease comprises cancer or autoimmune disease.

14. Use according to claim 12, characterized in that, The VAV1 -related disease comprises systemic lupus erythematosus, myasthenia gravis, periodontitis, type I diabetes, rheumatoid arthritis, multiple sclerosis, inflammatory enteritis, inflammatory bowel disease, autoimmune hepatitis, or psoriasis. The VAV1 -related disease comprises systemic lupus erythematosus, myasthenia gravis, periodontitis, type I diabetes, rheumatoid arthritis, multiple sclerosis, inflammatory enteritis, inflammatory bowel disease, autoimmune hepatitis, or psoriasis.

Images