Fused ring compounds, processes for their preparation and uses thereof

Abstract

The present invention relates to compounds of Formula X, stereoisomers, tautomers, or mixtures thereof, pharmaceutically acceptable salts, co-crystals, polymorphs, or solvates of the compounds, or stable isotopic derivatives, metabolites, or prodrugs of the compounds. As NLRP3 modulators (e.g., agonists or partial agonists), these compounds are useful in the treatment of diseases of abnormal cell proliferation (e.g., cancer).

Description

[0001] This application is a divisional application of the application with the application number 201910313420.8, the title of which is “Fused ring compounds, preparation methods and uses thereof”, the filing date of which is April 18, 2019. TECHNICAL FIELD

[0002] The present application relates to a new class of fused ring compounds, stereoisomers, tautomers or mixtures thereof, pharmaceutically acceptable salts, co-crystals, polymorphs or solvates of the compounds, or stable isotope derivatives, metabolites or prodrugs of the compounds. The compounds of the present application are useful as NLRP3 modulators (e.g., agonists or partial agonists) for the treatment of diseases of abnormal cell proliferation (e.g., cancer). BACKGROUND

[0003] NLRP3 (NLR family pyrin domain containing 3) belongs to the NOD-like receptor family and is one of the most studied intracellular pattern recognition receptors in recent years. It is mainly expressed in macrophages and neutrophils and is involved in the body's innate immunity against pathogen infection and stress damage. NLRP3 inflammasome plays a very important role in inflammatory and metabolic diseases. Its overactivation can lead to immune diseases such as type 2 diabetes, rheumatoid arthritis and atherosclerosis. However, recent studies have shown that NLRP3 has an anti-tumor effect of inhibiting tumor growth and metastasis.

[0004] NLRP3 protein, after recognizing pathogen-associated molecular patterns (PAMPs) or endogenous damage-associated molecular patterns (DAMPs), its NOD domain oligomerizes and recruits proteins such as ASC and pro-caspase-1 to form a functional NLRP3 inflammasome. After pro-caspase-1 is cleaved and activated into caspase-1, caspase-1 cleaves a large number of pro-IL-1β and pro-IL-18 to convert them into active forms IL-1β and IL-18 and release them into the extracellular space, amplifying the inflammatory response. The excited NLRP3 inflammasome can significantly increase the levels of immune factors IL-1β and IL-18 in the tumor microenvironment, initiating natural immune killing and subsequent acquired immune response to exert its anti-tumor effect. Specifically, IL-1β can induce CD8+T cells to secrete interferon gamma (IFN-γ) and CD4+cells to secrete IL-17, resulting in effective anti-tumor immune effect; while IL-18 can promote NK cell maturation, activate the STAT1 downstream signaling pathway in immune cells, and enhance the killing function of immune cells. Clinical studies have shown that the down-regulation of NLRP3 is significantly negatively correlated with the prognosis of liver cancer patients. Preclinical studies have also shown that NLRP3-deficient mice have a higher incidence of colorectal tumors and more severe colorectal cancer liver metastasis. Therefore, NLRP3 plays an important role in the tumor microenvironment and can be used as a key target for tumor immunotherapy and a tumor prognosis marker.

[0005] Although NLRP3 agonists have the potential for tumor immunotherapy, only one compound is currently in clinical phase I research, and four published compound patent applications (WO2017184746, WO2017184735, WO2018152396, WO2019014402). Therefore, there is a need to develop new, efficient and low-toxic NLRP3 agonists to meet the clinical treatment needs. SUMMARY

[0006] The inventors of the present application have, through creative labor, obtained a new class of fused ring compounds that can be used as NLRP3 modulators (e.g., agonists) to directly bind or modify NLRP3 at the protein level, thereby enhancing the function of NLRP3 inflammasome by activating, stabilizing, changing the distribution of NLRP3, or other means, thereby providing the following invention:

[0007] In one aspect, the present application provides a compound having the structure shown in Formula X, a stereoisomer, tautomer, or mixture thereof, a pharmaceutically acceptable salt, co-crystal, polymorph, or solvate of the compound, or a stable isotope derivative, metabolite, or prodrug of the compound:

[0008]

[0009] wherein:

[0010] X 2 is C, N, O or S, and satisfies the following conditions:

[0011] (1) when X 2 is O or S, R 2 and R 6 are absent;

[0012] (2) when X 2 is N, R 2 and R 6 are not both present;

[0013] R 6 is selected from H, C 1-6 alkyl, C 3-8 cycloalkyl, which C 1-6 alkyl and C 3-8 cycloalkyl can be optionally substituted by one or more of halogen, OH, CN, NO2, C 1-4 alkoxy, C 1-4 hydroxyalkyl;

[0014] R 2 is selected from H, C 1-8 alkyl, C 2-8 heteroalkyl, C 1-4 hydroxyalkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 6-12 aryl, C 3-8 cycloalkyl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl, which C 1-8 alkyl, C 2-8 heteroalkyl, C 1-4 hydroxyalkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 6-12 aryl, C 3-8 cycloalkyl, 5-10 membered heteroaryl, 4-10 membered heterocyclyl can be optionally substituted by one or more of halogen, C 1-4 haloalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 4-7 membered heterocyclyl, CN, NO2, OR 37 , SR 37 , C(O)R 30 , C(O)NR 31 R 32 , NR 33 C(O)R 34 , C(O)OR30 OC(O)R 30 OC(O)NR 31 R 32 NR 33 C(O)NR 31 R 32 NR 31 R 32 ;

[0015] R 3 H, halogen, CN, NO2, C 1-8 alkyl, C 1-8 alkoxy, C 2-8 heteroalkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, 9-12 membered arylperheterocyclyl, 9-12 membered arylperheteroaryl, 9-12 membered arylpercycloalkyl, 9-12 membered heteroarylpercycloalkyl, CO2R 30 , C(O)NR 31 R 32 , NR 33 C(O)R 34 , NR 31 R 32 , OR 37 , SR 37 , C(O)R 30 , OC(O)R 30 , OC(O)NR 31 R 32 , NR 33 C(O)NR 31 R 32 , NR 33 C(O)OR 30 , C(=NR 38 )NR 31 R 32 , NR 33 C(=NR 38 )NR 31 R 32 , P(R 39 )2, P(OR 39 )2, P(O)R 39 R 40 , P(O)OR 39 OR 30 , S(O)R 35 , S(O)2R 35 , S(O)NR 31 R 32 , S(O)2NR 31 R 32 ; said C1-8 Alkyl, C 1-8 Alkoxy, C 2-8 Heteroalkyl, C 3-8 Cycloalkyl, 4-10 membered heterocyclic groups, C 6-12 Aryl, 5-10-membered heteroaryl, 9-12-membered aryl-heterocyclic, 9-12-membered aryl-heteroaryl, and 9-12-membered aryl-cycloalkyl groups may optionally be substituted by one or more of the following substituents: halogen, CN, NO2, C. 1-4 Alkyl, C 3-8 cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy groups, C 1-4 Hydroxyalkyl, 4-10 membered heterocyclic group, C 6-12 Aryl, 5-10 aryl, 9-12 aryl heterocyclic, CO2R 30 C(O)R 30 C(O)NR 31 R 32 NR 33 C(O)R 34 NR 31 R 32 S(O)R 35 S(O)2R 35 S(O)NR 31 R 32 S(O)2NR 31 R 32 OR 37 SR 37 OC(O)R 30 OC(O)NR 31 R 32 NR 33 C(O)NR 31 R 32 NR 33 C(O)OR 30 C(=NR) 38 )NR 31 R 32 NR 33 C(=NR 38 )NR 31 R 32 =NNR 31 R 32 、P(R 39 2. P(OR) 39 2. P(O)R 39 R 40 P(O)OR 39 OR 30 ;

[0016] R 4 selected from C 1-15 alkyl, C 1-8 alkoxy, C 3-8 cycloalkyl; said C 1-15 alkyl, C 1-8 alkoxy, C 3-8 cycloalkyl can be optionally substituted by one or more of halogen, OH, CN, NO2, C 1-6 alkyl, C 1-4 haloalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy;

[0017] L is -(L 1 ) n -(L 2 ) p -(L 3 ) q - wherein L 1 , L 2 and L 3 are the same or different and each is independently selected from C 1-8 alkylene, C 2-8 alkenylene, C 2-8 alkynylene, C 1-8 alkyleneoxy, C 1-8 hydroxyalkylene, C 2-8 heteroalkylene, C 3-8 cycloalkylene, 4-10 membered heterocyclylene, C 6-12 arylene, 5-10 membered heteroarylene, O, S, NR 33 , SO, SO2, CO, C(R 36a R 36b ); said C 1-8 alkylene, C 2-8 alkenylene, C 2-8 alkynylene, C 1-8 alkyleneoxy, C 1-8 hydroxyalkylene, C 2-8 heteroalkylene, C 3-8 cycloalkylene, 4-10 membered heterocyclylene, C 6-12 arylene, 5-10 membered heteroarylene can be optionally substituted by one or more of halogen, OH, CN, NO2, C 1-6 alkyl, C 1-4 haloalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy, NR 31 R 32 ;

[0018] n, p, q are each independently selected from 0, 1 or 2; when n is 2, the two L 1 may be the same or different; when p is 2, the two L 2 may be the same or different; when q is 2, the two L 3 may be the same or different;

[0019] is selected from (I) wherein R 1 is selected from C 1-8 alkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, 9-12 membered arylheterocyclyl; said C 1-8 alkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, 9-12 membered arylheterocyclyl can be optionally substituted with one or more of the following substituents: halo, CN, NO2, C 1-4 alkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 hydroxyalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, 9-12 membered arylheterocyclyl, CO2R 30 , C(O)R 30 , C(O)NR 31 R 32 , NR 33 C(O)R 34 , NR 31 R 32 , OC(O)R 30 , OC(O)NR 31 R 32 , NR 33 C(O)NR 31 R 32 , NR 33 C(O)OR 30 , S(O)NR 31 R 32 , S(O)2NR 31 R 32 , S(O)R 35 , S(O)2R 35 , OR 37 , SR 37 ;

[0020] R 5each independently selected from H, halogen, C 1-6 alkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, said C 1-6 alkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy, C 3-8 cycloalkyl, 4-10 membered heterocyclyl can be optionally substituted with one or more of halogen, OH, CN, C 1-4 alkoxy, C 1-4 hydroxyalkyl, NR 31 R 32 ;

[0021] m is 0, 1 or 2;

[0022] and (II) wherein R 1 is as defined for formula (I); X 1 is CR 7 or N, R 7 is selected from H, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 2-8 heteroalkyl, C 3-8 cycloalkyl, 4-7 membered heterocyclyl, said C 1-6 alkyl, C 1-6 alkoxy, C 2-8 heteroalkyl, C 3-8 cycloalkyl, 4-7 membered heterocyclyl can be optionally substituted with one or more of halogen, OH, CN, C 1-4 alkoxy, C 1-4 hydroxyalkyl;

[0023] R 30 , R 37 , R 39 , R 40 each independently selected from hydrogen, C 1-8 alkyl, C 1-8 hydroxyalkyl, C 1-8 haloalkyl, C 1-8 alkoxy, C 1-8 haloalkoxy, C 2-8 heteroalkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, C 1-8 alkyl-C 6-12 aryl, C 1-8 alkyl-(5-10 membered heteroaryl); said C 1-8 alkyl, C 1-8hydroxyalkyl, C 1-8 haloalkyl, C 1-8 alkoxy, C 1-8 haloalkoxy, C 2-8 heteroalkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, C 1-8 alkyl-C 6-12 aryl, C 1-8 alkyl-(5-10 membered heteroaryl) can be optionally substituted with one or more of the following substituents: OH, CN, NO2, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 haloalkyl, halo, C 1-4 haloalkoxy, CO2(C 1-6 alkyl), CONR 31 R 32 , NR 31 R 32 , NR 33 C(O)R 34 , S(O)R 35 , S(O)2R 35 , S(O)NR 31 R 32 , S(O)2NR 31 R 32 ;

[0024] R 31 , R 32 , R 33 , R 34 are each independently selected from H, C 1-8 alkyl, C 1-8 hydroxyalkyl, C 1-8 alkoxy, C 2-8 heteroalkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl; or R 31 and R 32 together with the N atom to which they are attached form a 3-8 membered heterocyclyl; or R 33 and R 34 together with the C or N atom to which they are each attached form a 4-8 membered heterocyclyl; said C 1-8 alkyl, C 1-8 hydroxyalkyl, C 1-8 alkoxy, C 2-8 heteroalkyl, C 3-8 cycloalkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 6-12aryl, 5-10 membered heteroaryl, can be optionally substituted with one or more of the following: OH, CN, NO2, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 hydroxyalkyl, C 1-4 haloalkyl, C 1-4 haloalkoxy, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl;

[0025] R 35 selected from C 1-8 alkyl, C 1-8 hydroxyalkyl, C 1-8 haloalkyl, C 1-8 alkoxy, C 1-8 haloalkoxy, C 2-8 heteroalkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, C 1-8 alkyl-C 6-12 aryl, C 1-8 alkyl-(5-10 membered heteroaryl); said C 1-8 alkyl, C 1-8 hydroxyalkyl, C 1-8 haloalkyl, C 1-8 alkoxy, C 1-8 haloalkoxy, C 2-8 heteroalkyl, C 3-8 cycloalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, can be optionally substituted with one or more of the following: OH, CN, NO2, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 haloalkyl, halo, C 1-4 haloalkoxy, CO2(C 1-6 alkyl), CONR 31 R 32 , NR 31 R 32 , NR 33 C(O)R 34 , S(O)Me, S(O)2Me, S(O)NR 31 R 32 , S(O)2NR 31 R 32 ; wherein, R 31 , R 32 , R 33 , R 34 are as defined above;

[0026] R 36a and R 36b are the same or different, each independently selected from H, C 1-6 alkyl, C 1-6 alkoxy, C 1-8 hydroxyalkyl, C 1-8 haloalkyl; said C 1-6 alkyl, C 1-6 alkoxy, C 1-8 hydroxyalkyl, C 1-8 haloalkyl can optionally be substituted by one or more of the following: OH, CN, NH2, NHCH3, N(CH3)2; or R 36a and R 36b are connected, together with the carbon atom to which they are attached, form a 3-7 membered cycloalkyl or 4-7 membered heterocyclyl;

[0027] R 38 is selected from H, OH, CN, NO2, S(O)R 35 and S(O)2R 35 ;

[0028] when a plurality of R 30 are present, each R 30 may be the same or different;

[0029] when a plurality of R 31 are present, each R 31 may be the same or different;

[0030] when a plurality of R 32 are present, each R 32 may be the same or different;

[0031] when a plurality of R 33 are present, each R 33 may be the same or different;

[0032] when a plurality of R 34 are present, each R 34 may be the same or different;

[0033] when a plurality of R 35 are present, each R 35 may be the same or different;

[0034] when a plurality of R 37 are present, each R 37 may be the same or different;

[0035] when a plurality of R 38 are present, each R 38 may be the same or different;

[0036] when multiple R 39 each R 39 may be the same or different;

[0037] when multiple R 40 each R 40 may be the same or different.

[0038] In certain embodiments, R 1 is selected from C 3-6 cycloalkyl, 4-7 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl; said C 3-6 cycloalkyl, 4-7 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl can be optionally substituted with one or more of the following substituents: halo, CN, C 1-4 alkyl, C 3-6 cycloalkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 hydroxyalkyl;

[0039] In certain embodiments, R 2 is selected from H, C 1-8 alkyl, C 2-8 heteroalkyl, C 1-4 hydroxyalkyl, said C 1-8 alkyl, C 2-8 heteroalkyl, C 1-4 hydroxyalkyl can be optionally substituted with one or more of the following substituents: halo, C 1-4 haloalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, 4-7 membered heterocyclyl, CN, OR 37 , SR 37 , C(O)R 30 , C(O)NR 31 R 32 , NR 33 C(O)R 34 , C(O)OR 30 , OC(O)R 30 , OC(O)NR 31 R 32 , NR 33 C(O)NR 31 R 32 , NR 31 R 32 ;

[0040] In certain embodiments, R 3 is selected from H, halo, CN, C1-6 alkyl, C 1-6 alkoxy, C 2-6 heteroalkyl, C 3-6 cycloalkyl, 4-8 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, 9-12 membered arylheterocyclyl, 9-12 membered arylheteroaryl, 9-12 membered arylcycloalkyl, 9-12 membered heteroarylcycloalkyl, -CO2R 30 , -C(O)R 30 , -C(O)NR 31 R 32 , -NR 33 C(O)R 34 , -NR 31 R 32 , NR 33 C(O)NR 31 R 32 , -S(O)2R 35 , -OR 37 ; said C 1-6 alkyl, C 1-6 alkoxy, C 2-6 heteroalkyl, C 3-6 cycloalkyl, 4-8 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, 9-12 membered arylheterocyclyl, 9-12 membered arylheteroaryl, 9-12 membered arylcycloalkyl, or 9-12 membered heteroarylcycloalkyl can be optionally substituted with one or more of the following substituents: halo, CN, C 1-4 alkyl, C 3-8 cycloalkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 1-4 hydroxyalkyl, 4-10 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl, 9-12 membered arylheterocyclyl, CO2R 30 , C(O)R 30 , C(O)NR 31 R 32 , NR 33 C(O)R 34 , NR 31 R 32 , S(O)2R 35 , S(O)NR 31 R 32 , S(O)2NR 31 R 32 , OR 37 , SR 37 , OC(O)R 30 , OC(O)NR31 R 32 , NR 33 C(O)NR 31 R 32 , NR 33 C(O)OR 30 ;

[0041] In certain embodiments, R 4 is selected from C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl; which C 1-4 alkyl, C 1-4 alkoxy, or C 3-6 cycloalkyl can be optionally substituted with one or more of halogen, OH, CN, C 1-6 alkyl, C 1-4 haloalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy;

[0042] In certain embodiments, R 7 is selected from H, halogen, C 1-4 alkyl, C 3-6 cycloalkyl, C 2-6 heteroalkyl, 4-7 membered heterocyclyl, which C 1-4 alkyl, C 3-6 cycloalkyl, C 2-6 heteroalkyl, or 4-7 membered heterocyclyl can be optionally substituted with one or more of halogen, OH, CN, C 1-4 alkoxy, C 1-4 hydroxyalkyl.

[0043] In certain embodiments, R 5 is selected from halogen, C 1-4 alkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, 4-7 membered heterocyclyl, which C 1-4 alkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, or 4-7 membered heterocyclyl can be optionally substituted with one or more of halogen, OH, CN, C 1-4 alkoxy, C 1-4 hydroxyalkyl, NR 31 R 32 ; and when multiple R 5 are present, each R 5 can be the same or different;

[0044] In certain embodiments, L is -(L 1 ) n -(L 2 ) p -(L 3 ) q - wherein L 1 , L 2 and L 3 are the same or different and each is independently selected from C 1-8 alkylene, C 1-4 alkylenoxy, C 1-4 hydroxyalkylene, C 2-6 heteroalkylene, C 3-6 cycloalkylene, 4-7 membered heterocyclylene, phenylene, 5-6 membered heteroarylene, O, S, -NR 33 , SO, SO2, CO, -C(R 36a R 36b ); said C 1-8 alkylene, C 1-4 alkylenoxy, C 1-4 hydroxyalkylene, C 2-6 heteroalkylene, C 3-6 cycloalkylene, 4-7 membered heterocyclylene, 5-6 membered heteroarylene can be optionally substituted with one or more of halogen, OH, CN, C 1-6 alkyl, C 1-4 haloalkyl, C 1-4 hydroxyalkyl, C 1-4 alkoxy, NR 31 R 32 ;

[0045] wherein R 30 , R 31 , R 32 , R 33 , R 34 , R 35 , R 37 , R 36a , R 36b are as defined above.

[0046] In certain embodiments, X 1 is CR 7 , R 7 is selected from H, halogen, C 1-4 alkyl, C 2-6 heteroalkyl, optionally said C 1-4 alkyl or C 2-6 heteroalkyl is substituted with one or more halogen.

[0047] In certain embodiments, X 2 is N or C, preferably N, and R2 H, C 1-8 alkyl, C 2-8 heteroalkyl, C 1-4 hydroxyalkyl, said C 1-8 alkyl, C 2-8 heteroalkyl, or C 1-4 hydroxyalkyl can be optionally substituted with one or more of halogen, C 1-4 hydroxyalkyl, C 1-4 alkoxy, 4-7 membered heterocyclyl, OR 37 , NR 31 R 32 .

[0048] In certain embodiments, R 2 is selected from H, C 1-4 alkyl, C 2-6 heteroalkyl, C 1-4 hydroxyalkyl, said C 1-4 alkyl, C 2-6 heteroalkyl or C 1-4 hydroxyalkyl can be optionally substituted with one or more of halogen, C 1-4 hydroxyalkyl, C 1-4 alkoxy, 4-7 membered heterocyclyl, OR 37 , NR 31 R 32 .

[0049] In certain embodiments, R 1 is selected from C 6-12 aryl and 5-10 membered heteroaryl, said C 6-12 aryl, 5-10 membered heteroaryl can be optionally substituted with one or more of halogen, CN, C 1-4 alkyl, C 3-6 cycloalkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 hydroxyalkyl;

[0050] In certain embodiments, R 1 is selected from phenyl and 5-6 membered heteroaryl, said phenyl or 5-6 membered heteroaryl can be optionally substituted with one or more of halogen, -CN, C 1-4 alkyl, C 3-6 cycloalkyl, C 1-4 haloalkyl, -C 1-4 alkoxy, -C 1-4 haloalkoxy, -C 1-4 hydroxyalkyl.

[0051] In certain embodiments, R 3 is selected from H, CN, C1-6 alkyl, C 3-6 cycloalkyl, 4-8 membered heterocyclyl, phenyl, 5-6 membered heteroaryl, -CO2R 30 , -C(O)R 30 , -S(O)2R 35 , -OR 37 , -NR 31 R 32 , NR 33 C(O)NR 31 R 32 , NR 33 C(O)R 34 ; said C 1-6 alkyl, C 3-6 cycloalkyl, 4-8 membered nitrogen-containing heterocyclyl, 4-8 membered oxygen-containing heterocyclyl, phenyl, 5-6 membered heteroaryl can be optionally substituted with one or more of the following substituents: C 1-4 alkyl, halogen, CN, C 3-8 cycloalkyl, C 1-3 alkoxy, OH, -S(O)2R 35 , -C(O)R 30 , -NR 31 R 32 , NR 33 C(O)NR 31 R 32 or NR 33 C(O)R 34 .

[0052] In certain embodiments, R 4 is selected from C 1-4 alkyl, C 3-6 cycloalkyl, said C 1-4 alkyl or C 3-6 cycloalkyl can be optionally substituted with one or more halogen.

[0053] In certain embodiments, R 5 is selected from halogen, C 1-4 alkyl, and m is 0 or 1.

[0054] In certain embodiments, L is -(L 1 ) n -(L 2 ) p -(L 3 ) q -, wherein L 1 , L 2 and L 3 are the same or different, and each is independently selected from C 1-4 alkylene, C 2-6heteroalkylene, 4-7 membered heterocyclylene, O, S, NR 33 C 1-4 alkylene, CO; said C 1-4 alkylene, C 2-6 heteroalkylene, 4-7 membered heterocyclylene, or C 1-4 alkylene can be optionally substituted with one or more of the following substituents: halo, OH, NR 31 R 32 .

[0055] n, p, q are each independently 0, 1, or 2.

[0056] In certain embodiments, R 30 is selected from H and C 1-4 alkyl; preferably, R 30 is selected from H, methyl, ethyl, propyl, and t-butyl.

[0057] In certain embodiments, R 37 is selected from H, C 1-8 alkyl (e.g., C 1-4 alkyl), 4-10 membered heterocyclyl (e.g., 4-7 membered heterocyclyl); said C 1-8 alkyl or 4-10 membered heterocyclyl can be optionally substituted with one or more of the following substituents: OH, CONR 31 R 32 , NR 31 R 32 , NR 33 C(O)R 34 ; in certain embodiments, R 37 is selected from H, methyl, ethyl, propyl, and t-butyl; wherein, R 31 , R 32 , R 33 , R 34 are as defined above.

[0058] In certain embodiments, R 31 , R 32 , R 33 , R 34 are each independently selected from H, C 1-8 alkyl (e.g., C 1-4 alkyl), C 1-8 hydroxyalkyl (e.g., C 1-4 hydroxyalkyl), C 1-8 alkoxy (e.g., C 1-4 alkoxy), 4-10 membered heterocyclyl (e.g., 4-7 membered heterocyclyl); said C 1-8 alkyl, C 1-8 hydroxyalkyl, C 1-8alkyl, C 1-4 alkyl, C 1-4 alkyl, C 1-4 alkyl, C 1-4 haloalkyl, C 33 haloalkyl, C 1-4 haloalkyl, C 33 haloalkyl, C

[0059] haloalkyl, C 31 haloalkyl, C 32 haloalkyl, C 33 haloalkyl, C 34 haloalkyl, C 1-4 haloalkyl, C 1-4 haloalkyl, C 1-4 haloalkyl, C 1-4 haloalkyl, C

[0060] haloalkyl, C 35 haloalkyl, C 1-8 haloalkyl, C 1-4 haloalkyl, C 1-8 haloalkyl, C 1-4 haloalkyl, C 31 haloalkyl, C 32 haloalkyl, C 35 haloalkyl, C

[0061] haloalkyl, C 36a haloalkyl, C 36b haloalkyl, C 1-6 haloalkyl, C 1-4 haloalkyl, C 1-6 haloalkyl, C 36a haloalkyl, C 36b haloalkyl, C 3-6cycloalkyl or 4-7 membered heterocyclyl, said C 3-6 Cycloalkyl or 4-7 membered heterocyclyl can be optionally substituted with one or more of OH, NH2, NHCH3, N(CH3)2.

[0062] In certain embodiments, R 38 is selected from H, CN and S(O)2R 35 .

[0063] In certain embodiments, X 1 is CH.

[0064] In certain embodiments, X 2 is N, R 6 is absent, R 2 is H or C 1-4 alkyl.

[0065] In certain embodiments, R 1 is selected from phenyl, pyrrolyl, oxazolyl, imidazolyl, pyrazolyl, pyridinyl and pyrimidinyl. In certain embodiments, R 1 is pyrazolyl, for example 1H-pyrazol-3-yl, 1H-pyrazol-1-yl.

[0066] In certain embodiments, R 2 is selected from H, methyl, ethyl, propyl, butyl, hydroxymethyl, hydroxyethyl, hydroxypropyl and hydroxybutyl, 2-methoxyethyl, 2-methoxypropyl.

[0067] In certain embodiments, R 3 is selected from H, C 1-4 alkyl, cyclopropyl, 4-7 membered heterocyclyl (for example piperazinyl, morpholinyl, piperidinyl, pyrrolidinyl, tetrahydrofuranyl, azetidinyl), phenyl, 5-6 membered heteroaryl (for example pyridinyl, pyrazolyl), OR 37 ; said C 1-4 alkyl, cyclopropyl, 4-7 membered nitrogen-containing heterocyclyl, phenyl, 5-6 membered heteroaryl can be optionally substituted with one or more (for example 1, 2, 3 or 4) methyl, F, Cl, CN, C 1-3 alkoxy, OH, S(O)2R 35 or C(O)R 30 .

[0068] In certain embodiments, L is -(L 1 ) n -(L 2 ) p -(L 3 ) q -, wherein L 1 , L 2 and L3 the same or different and each independently selected from the group consisting of methylene, ethylene, propylene, O, NR 33 , C 1-4 alkyleneoxy, CO.

[0069] In certain embodiments, R 4 is selected from the group consisting of methyl, isopropyl, cyclopropyl, trifluoromethyl.

[0070] In certain embodiments, R 7 is selected from the group consisting of H, F, Cl, methyl, cyclopropyl.

[0071] In certain embodiments, R 5 is selected from the group consisting of F, Cl, C 1-4 alkyl.

[0072] In certain embodiments, m is 0 or 1 ; preferably, m is 0.

[0073] In certain embodiments, n, p, q are each independently 0, 1 or 2.

[0074] In certain embodiments, -L-R 3 is selected from the group consisting of:

[0075]

[0076] In some embodiments, the compounds of the present application have the structure according to Formula III or Formula IV:

[0077]

[0078] R 1 , R 3 , R 4 , R 5 , X 1 , L, m are as defined above for Formula X. When X 2 is O or S, R 2 is absent; when X 2 is N, R 2 is as defined above for Formula X.

[0079] In some embodiments, the compounds of the present application have the structure according to Formula V or Formula VI:

[0080]

[0081] R 1 , R 2 , R 3 , R 4 , R 5 , X 1 , L, m are as defined above for Formula X.

[0082] In some embodiments, the compound of the present application has the structure of Formula V-A or Formula VI-A:

[0083]

[0084] R 1 , R 2 , R 3 , R 4 , L is defined as above for Formula X.

[0085] In embodiments of the present application, the compound of the present application is selected from, but not limited to:

[0086]

[0087] In another aspect, the present application provides a pharmaceutical composition comprising a compound as described above, a stereoisomer, a tautomer, or a mixture thereof of the compound, a pharmaceutically acceptable salt, a co-crystal, a polymorph, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound. Optionally, the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers.

[0088] In some embodiments, the pharmaceutical composition is used for preventing, alleviating, and / or treating a disease associated with NLRP3 inflammasome activity (e.g., a cell proliferation abnormality disease, e.g., a tumor disease).

[0089] Optionally, the pharmaceutical composition of the present application further comprises one or more second therapeutic agents. In certain embodiments, the second therapeutic agent includes other drugs for treating diseases such as tumors.

[0090] In another aspect, the present application provides a pharmaceutical preparation comprising a compound as described above, a stereoisomer, a tautomer, or a mixture thereof of the compound, a pharmaceutically acceptable salt, a co-crystal, a polymorph, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, or a pharmaceutical composition as described above.

[0091] In another aspect, the present application provides the use of a compound as described above, a stereoisomer, a tautomer, or a mixture thereof of the compound, a pharmaceutically acceptable salt, a co-crystal, a polymorph, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, or a pharmaceutical composition as described above for the preparation of a medicament for preventing, alleviating, and / or treating a disease associated with NLRP3 inflammasome activity (e.g., a cell proliferation abnormality disease, e.g., a tumor disease).

[0092] In another aspect, the present application provides use of a compound as described above, a stereoisomer, a tautomer, or a mixture thereof of the compound, a pharmaceutically acceptable salt, a co-crystal, a polymorph, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, or a pharmaceutical composition as described above, for the preparation of a formulation for modulating (e.g., increasing) the activity of NLRP3 inflammasome.

[0093] In some embodiments, the formulation is administered to a subject (e.g., a mammal; e.g., a bovine, equine, ovine, porcine, canine, feline, rodent, primate; e.g., a human) in vivo to increase the NLRP3 inflammasome activity in cells in the subject; or, the formulation is administered to cells (e.g., a cell line or cells from a subject) in vitro to increase the NLRP3 inflammasome activity in the cells.

[0094] In another aspect, the present application provides a method of modulating (e.g., increasing) the activity of NLRP3 inflammasome in a cell, comprising administering to the cell an effective amount of a compound as described above, a stereoisomer, a tautomer, or a mixture thereof of the compound, a pharmaceutically acceptable salt, a co-crystal, a polymorph, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, or a pharmaceutical composition as described above, or a pharmaceutical formulation as described above.

[0095] In another aspect, the present application provides a kit for modulating (e.g., increasing) the activity of NLRP3 inflammasome, comprising a compound of the present application, a stereoisomer, a tautomer, or a mixture thereof of the compound, a pharmaceutically acceptable salt, a co-crystal, a polymorph, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, or a pharmaceutical composition as described above, or a formulation as described above.

[0096] In another aspect, the present application provides a method of preventing, alleviating, and / or treating a disease (e.g., a neoplastic disease) associated with the activity of NLRP3 inflammasome, comprising administering to a subject in need thereof a therapeutically and / or prophylactically effective amount of a compound of the present application, a stereoisomer, a tautomer, or a mixture thereof of the compound, a pharmaceutically acceptable salt, a co-crystal, a polymorph, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, or a pharmaceutical composition as described above, or a formulation as described above.

[0097] Optionally, the method further comprises administering to the subject in need thereof one or more second therapeutic agents. In some embodiments, the second therapeutic agent comprises other drugs for treating diseases such as neoplasms.

[0098] In the present invention, the tumor diseases include, but are not limited to, lung cancer, pancreatic cancer, breast cancer, head and neck cancer, liver cancer, melanoma, glioma.

[0099] In some embodiments, the compounds of the present invention are full agonists; in some embodiments, the compounds of the present invention are partial agonists.

[0100] Definitions of terms

[0101] Unless otherwise defined herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. References herein to technical terms used herein are intended to refer to the technical terms as commonly understood by those in the art, including variations or substitutions of techniques or replacements of equivalent techniques that would be apparent to those skilled in the art. Although the following terms are believed to be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain the present invention.

[0102] The term "agonist" refers to a compound that binds to a receptor and activates it to elicit a downstream biological effect or response, including full agonists and partial agonists. A full agonist can activate the receptor and produce a maximal effect or E max ). A partial agonist can bind to and activate the receptor, but produces a partial effect relative to the full agonist. When a full agonist and a partial agonist coexist, the partial agonist can sometimes act as a partial antagonist by competing with the full agonist for the binding site on the receptor or other mechanisms. The potency (measured by EC 50 ) of a partial agonist can be higher or lower than that of a full agonist. The NLRP3 agonists of the present invention include NLRP3 full agonists and NLRP3 partial agonists.

[0103] The term "NLRP3" is the full name of NLR family pyrin domain containing 3, which is an inflammasome. In the present invention, when referring to "NLRP3", the meaning includes nucleic acids, polynucleotides, oligonucleotides, sense and antisense polynucleotide strands, complementary sequences, short peptides, polypeptides, proteins, homologous or heterologous molecules, isoforms, precursors, mutants, variants, derivatives, various splice bodies, alleles, different species, and activated fragments, etc. of NLRP3.

[0104] The terms "comprising," "containing," "having," "including," "involving," and "including" and other similar forms are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0105] The term "halo" means substituted with a halogen atom, said "halogen" includes F, CI, Br, or I.

[0106] The term "alkyl" is a straight-chain or branched saturated aliphatic hydrocarbon group. The terms "C 1-15 alkyl," "C 1-8 alkyl," "C 1-6 alkyl," and "C 1-4 alkyl" mean a straight-chain or branched alkyl group having 1 to 15 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, and 1 to 4 carbon atoms, respectively, such as methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, or n-hexyl. The alkyl group can be optionally substituted with one or more (such as 1 to 3) same or different substituents.

[0107] The term "alkylene" means a saturated divalent hydrocarbon radical, straight-chained or branched, obtained by removing two hydrogen atoms from an alkyl group, containing the number of carbon atoms indicated. For example, alkylene of 1 to 8 carbon atoms, such as methylene (-CH2-), ethylene (-CH2CH2-), isopropylene (-CH(CH3)CH2-), and the like; the alkylene group can be optionally substituted with one or more (such as 1 to 3) same or different substituents.

[0108] The term "haloalkyl" means an alkyl group substituted with one or more (such as 1 to 3) same or different halogen atoms, the terms "C 1-8 haloalkyl," "C 1-6 haloalkyl," and "C 1-4 haloalkyl" mean a haloalkyl group having 1 to 8 carbon atoms, 1 to 6 carbon atoms, and 1 to 4 carbon atoms, respectively, such as -CF3, -C2F5, -CHF2, -CH2F, -CH2CF3, -CH2CI, or -CH2CH2CF3, and the like.

[0109] The term "hydroxyalkyl" means a group formed by the substitution of one or more hydrogen atoms of an alkyl group with one or more hydroxyl groups, such as C 1-4 hydroxyalkyl or C 1-3 hydroxyalkyl, examples of which include, but are not limited to, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, -CH(OH)CH3, and the like.

[0110] The term "alkenyl" means a monovalent straight or branched chain hydrocarbon group containing one or more carbon-carbon double bonds, such as -CH=CH2, -CH2CH=CH2, -C(CH3)=CH2, -CH2-CH=CH-CH3, and the like, which alkenyl group can be optionally substituted with one or more (such as 1 to 3) of the same or different substituents.

[0111] The term "alkenylene" means a divalent straight or branched chain aliphatic group containing one or more carbon-carbon double bonds, containing the specified number of carbon atoms, such as 2 to 8 carbon atoms, such as -CH=CH-, -CH2CH=CH-, -C(CH3)=CH-, and the like, which alkenylene group can be optionally substituted with one or more (such as 1 to 3) of the same or different substituents.

[0112] The term "alkynyl" means a monovalent straight or branched chain hydrocarbon group having one or more carbon-carbon triple bonds, including but not limited to ethynyl, 2- propynyl, 2-butynyl, and 1,3-butadiynyl, and the like, which alkynyl group can be optionally substituted with one or more (such as 1 to 3) of the same or different substituents.

[0113] The term "alkynylene" means a divalent straight or branched chain hydrocarbon group having one or more carbon-carbon triple bonds, containing the specified number of carbon atoms, such as 2 to 8 carbon atoms, including but not limited to and the like, which alkynylene group can be optionally substituted with one or more (such as 1 to 3) of the same or different substituents.

[0114] The term "alkoxy" means an group in which an oxygen atom is inserted between an alkyl group (as defined above) at any reasonable position, preferably C 1-8 alkoxy, C1-C6alkoxy, C1-C4alkoxy, or C1-C3alkoxy. Representative examples of C1-C6alkoxy include, but are not limited to, methoxy, ethoxy, propyloxy, isopropoxy, n- propoxy, isopropoxy, n-butoxy, isobutoxy, t-butoxy, pentoxy, hexyloxy, -CH2-OCH3, and the like, which alkoxy group can be optionally substituted with one or more (such as 1 to 3) of the same or different substituents.

[0115] The term "alkyleneoxy" means a divalent alkoxy group, such as -OCH2-, -OCH(CH3)CH2-, -OCH2CH2O-, -CH2CH2O-, and the like, which alkyleneoxy group can be optionally substituted with one or more (such as 1 to 3) of the same or different substituents.

[0116] The term "heteroalkyl" refers to an optionally substituted alkyl group having two carbon atoms or more, having one or more skeletal chain atoms selected from an atom other than carbon, e.g., oxygen, nitrogen, sulfur, phosphorus, or combinations thereof. The numerical ranges given are the number of carbons in the chain, e.g., C 2-8 Heteroalkyl groups contain 2 to 8 carbon atoms. For example, -CH2OCH2CH3, -CH2NHCH2CH3, or -CH2N(Me)CH2CH3is designated as a C3heteroalkyl group. The term "heteroalkylene" denotes the corresponding divalent radical, e.g., -CH2OCH2CH2-.

[0117] The term "fused ring" or "fused" refers to a ring system formed by two or more cyclic structures sharing two adjacent atoms with each other.

[0118] The term "spirocyclic" refers to a ring system formed by two or more cyclic structures sharing one ring atom with each other.

[0119] The term "bridged ring" refers to a ring system formed by two or more cyclic structures sharing two non-adjacent atoms with each other.

[0120] The term "cycloalkyl" refers to a saturated or unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring group, e.g., "C 3-8 "Cycloalkyl" refers to a cycloalkyl group having from 3 to 8 ring-forming carbon atoms, e.g., C 3-6 Cycloalkyl groups can be monocycloalkyl groups, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and bicycloalkyl groups, e.g., C 3-8 spirocycloalkyl, C 3-8 bridged cycloalkyl, C 3-8 fused cycloalkyl, C 3-6 spirocycloalkyl, C 3-6 bridged cycloalkyl, C 3-6 fused cycloalkyl. In the present application, cycloalkyl groups can be optionally substituted with one or more (such as 1 to 3) substituents, which can be the same or different. Carbon atoms on a cycloalkyl group are optionally oxidized (i.e., forming C=O).

[0121] The term "cycloalkylene" refers to a cycloalkyl group as defined herein having two monovalent radical centers obtained by removal of two hydrogen atoms from the same carbon atom or from two different carbon atoms of the parent cycloalkyl group. Typical cycloalkylene groups include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene, cyclooctylene, cyclononylene, cyclohexenylene, and the like.

[0122] The term "aryl" refers to an all-carbon monocyclic or fused ring polycyclic aromatic group having a conjugated π-electron system. As used herein, the term "C 6-12"Aryl" refers to an aryl group containing 6 to 12 carbon atoms, such as phenyl or naphthyl. The aryl group may optionally be substituted by one or more (such as 1 to 3) identical or different substituents (e.g., halogen, OH, CN, NO2, C1-C6 alkyl, etc.).

[0123] The term "arylene" refers to an aryl group as defined herein, having two monovalent group centers obtained by removing two hydrogen atoms from the same carbon atom or two different carbon atoms of the parent aryl group. Typical arylene groups include, but are not limited to, phenylene and naphthylene.

[0124] The term "aryl cycloalkyl" refers to a cyclic group formed by an aryl group and a cycloalkyl group (e.g., a monocycloalkyl group) sharing two adjacent atoms, wherein the connection point with other groups can be on the aryl group or on the cycloalkyl group. The term "9-12-membered aryl cycloalkyl" refers to an aryl cycloalkyl group containing a total of 9-12 ring atoms, such as phenylcyclopentyl or phenylcyclohexyl.

[0125] The term "heterocyclic group" refers to a monocyclic or polycyclic (e.g., fused, spirocyclic, or bridged) group having two or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) carbon atoms and one or more (e.g., 1, 2, 3, or 4) heteroatoms, including but not limited to oxygen, nitrogen, and sulfur atoms, wherein the carbon atoms and heteroatoms on the heterocyclic group are optionally oxidized (oxo) (e.g., forming C=O, S(=O) or S(=O)2). For example, "3-14 membered heterocyclic group" refers to a heterocyclic group containing 3-14 ring atoms, including but not limited to 4-10 membered heterocyclic groups, 4-7 membered heterocyclic groups, 5-6 membered heterocyclic groups, 4-7 membered nitrogen-containing heterocyclic groups, 4-7 membered oxygen-containing heterocyclic groups, 4-7 membered sulfur-containing heterocyclic groups, 5-6 membered nitrogen-containing heterocyclic groups, 5-6 membered oxygen-containing heterocyclic groups, and 5-6 membered sulfur-containing heterocyclic groups. The "nitrogen-containing heterocyclic group," "oxygen-containing heterocyclic group," and "sulfur-containing heterocyclic group" optionally also contain one or more other heteroatoms selected from oxygen, nitrogen, and sulfur. Examples of 3-14 membered heterocyclic groups include, but are not limited to, ethylene oxide, aziridinyl, aziridine, oxadiazinyl, tetrahydrofuranyl, pyrrolylalkyl, pyrrolidone, imidazoalkyl, pyrazolylalkyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazine, and trithianyl. wait

[0126] The term “subheterocyclic group” refers to a heterocyclic group as defined herein, which has two monovalent group centers obtained by removing two hydrogen atoms from the same carbon atom or two different carbon atoms, one carbon atom and one heteroatom, or two heteroatoms.

[0127] In the present application, the heterocyclyl group can form a fused ring structure with a heterocyclyl group or a cycloalkyl group, and the point of attachment of the fused ring structure to other groups can be on either the heterocyclyl group or the cycloalkyl group. Thus, the heterocyclyl group of the present application also includes, but is not limited to, heterocyclyl fused heterocyclyl, heterocyclyl fused cycloalkyl, mono-heterocyclyl fused mono-heterocyclyl, mono-heterocyclyl fused mono-cycloalkyl, for example, 3-7 membered (mono)heterocyclyl fused 3-7 membered (mono)heterocyclyl, 3-7 membered (mono)heterocyclyl fused (mono)cycloalkyl, 3-7 membered (mono)heterocyclyl fused C 4-6 mono-heterocyclyl fused mono-cycloalkyl, for example, 3-7 membered (mono)heterocyclyl fused 3-7 membered (mono)heterocyclyl, 3-7 membered (mono)heterocyclyl fused (mono)cycloalkyl, 3-7 membered (mono)heterocyclyl fused C

[0128] In the present application, the heterocyclyl group also includes bridged heterocyclyl and spiroheterocyclyl.

[0129] The term "bridged heterocycle" refers to a cyclic structure containing one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen atoms, nitrogen atoms, sulfur atoms) formed by two saturated rings sharing two non-adjacent ring atoms, including but not limited to 7-10 membered bridged heterocycles, 8-10 membered bridged heterocycles, 7-10 membered nitrogen-containing bridged heterocycles, 7-10 membered oxygen-containing bridged heterocycles, 7-10 membered sulfur-containing bridged heterocycles, and the like, for example The "nitrogen-containing bridged heterocycle", "oxygen-containing bridged heterocycle", "sulfur-containing bridged heterocycle" optionally further contains one or more additional heteroatoms selected from oxygen, nitrogen, sulfur.

[0130] The term "spiroheterocycle" refers to a cyclic structure containing one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen atoms, nitrogen atoms, sulfur atoms) formed by two or more saturated rings sharing one ring atom, including but not limited to 5-10 membered spiroheterocycles, 6-10 membered spiroheterocycles, 6-10 membered nitrogen-containing spiroheterocycles, 6-10 membered oxygen-containing spiroheterocycles, 6-10 membered sulfur-containing spiroheterocycles, and the like, for example The "nitrogen-containing spiroheterocycle", "oxygen-containing spiroheterocycle", "sulfur-containing spiroheterocycle" optionally further contains one or more additional heteroatoms selected from oxygen, nitrogen, sulfur. The term "6-10 membered nitrogen-containing spiroheterocycle" refers to a spiroheterocycle containing a total of 6-10 ring atoms and at least one of which is a nitrogen atom.

[0131] The term "aryl and heterocyclyl" refers to cyclic groups formed by aryl and heterocyclyl groups sharing two adjacent carbon atoms with each other, the point of attachment to other groups being on either the aryl or heterocyclyl group. Wherein aryl and heterocyclyl are as defined above. For example, as used herein, the term "9-12 membered aryl and heterocyclyl" means groups of aryl and heterocyclyl groups containing a total of 9-12 ring atoms, including but not limited to 9-10 membered benzo-heterocyclyl groups, for example, phenyl and 5-8 membered heterocyclyl groups, for example, phenyl and 5-6 membered heterocyclyl groups, for example, benzo 5-6 membered monocyclic heterocyclyl groups, benzo 5-6 membered nitrogen-containing monocyclic heterocyclyl groups, benzo 5-6 membered oxygen-containing monocyclic heterocyclyl groups, benzo 5-6 membered sulfur-containing monocyclic heterocyclyl groups, the "nitrogen-containing heterocyclyl groups", "oxygen-containing heterocyclyl groups", "sulfur-containing heterocyclyl groups" optionally further containing one or more additional heteroatoms selected from oxygen, nitrogen, sulfur. The carbon atoms and heteroatoms on the heterocyclyl groups are optionally oxo (e.g., forming C=0, S(=0), or S(=0)2).

[0132] Examples include, but are not limited to: indazolyl,

[0133] The term "heteroaryl" refers to monocyclic or polycyclic aromatic groups containing one or more heteroatoms which are the same or different, including monocyclic heteroaryl groups and bicyclic or polycyclic ring systems containing at least one heteroaromatic ring (an aromatic ring system containing at least one heteroatom), which can have, for example, 5, 6, 7, 8, 9, or 10 ring atoms. The heteroatoms can be oxygen, nitrogen, or sulfur. The carbon atoms and heteroatoms on the heteroaryl groups can be optionally oxo (e.g., forming C=0, S(=0), or S(=0)2). For example, "5-10 membered heteroaryl" means heteroaryl groups containing 5 to 10 ring atoms, including 5-6 membered heteroaryl groups, 5-6 membered monocyclic heteroaryl groups, 5-10 membered nitrogen-containing heteroaryl groups, 5-10 membered oxygen-containing heteroaryl groups, 5-10 membered sulfur-containing heteroaryl groups, 5-6 membered nitrogen-containing heteroaryl groups, 5-6 membered oxygen-containing heteroaryl groups, 5-6 membered sulfur-containing heteroaryl groups, 5-6 membered nitrogen-containing monocyclic heteroaryl groups, 5-6 membered oxygen-containing monocyclic heteroaryl groups, 5-6 membered sulfur-containing monocyclic heteroaryl groups. The "nitrogen-containing heteroaryl groups", "oxygen-containing heteroaryl groups", "sulfur-containing heteroaryl groups", "nitrogen-containing monocyclic heteroaryl groups", "oxygen-containing monocyclic heteroaryl groups", "sulfur-containing monocyclic heteroaryl groups" optionally further contain one or more additional heteroatoms selected from oxygen, nitrogen, sulfur. Examples include, but are not limited to, thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, triazolyl, tetrazolyl, oxadiazolyl, thiadiazolyl, and the like, or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and the like, as well as 5-10 membered annulated groups containing these groups.

[0134] The term "heteroarylene" refers to a divalent radical of a heteroaryl group as defined above, obtained by the removal of two hydrogen atoms from a single carbon atom or two different carbon atoms of the parent heteroaryl group, or one hydrogen atom from a carbon atom and one hydrogen atom from a nitrogen atom of the parent heteroaryl group.

[0135] In the present application, a heteroaryl group (e.g., a mono-heteroaryl group) can share two adjacent atoms with an aryl group (e.g., a monocyclic aryl group, e.g., phenyl), a heterocyclyl group (e.g., a monocyclic heterocyclyl group), a cycloalkyl group (e.g., a monocyclic cycloalkyl group), or another heteroaryl group (e.g., another mono-heteroaryl group) to form a fused ring structure, which can be attached at any of the heteroaryl rings or other rings, including but not limited to (mono)heteroaryl-(mono)heteroaryl, (mono)heteroaryl-(mono)cycloalkyl, (mono)heteroaryl-(mono)cycloalkyl, and (mono)heteroaryl-(mono)cycloalkyl, e.g., 5-6 membered (mono)heteroaryl fused with 5-6 membered (mono)heteroaryl, 5-6 membered (mono)heteroaryl fused with phenyl, 5-6 membered (mono)heteroaryl fused with 5-6 membered (mono)heterocyclyl, or 5-6 membered (mono)heteroaryl fused with C 4-6 (monocyclic) cycloalkyl, e.g., 5-6 membered heteroaryl fused with cyclobutyl, 5-6 membered heteroaryl fused with cyclopentyl, 5-6 membered heteroaryl fused with cyclohexyl, and the like.

[0136] The term "aryl-heteroaryl" refers to a fused ring group of an aryl group (e.g., a monocyclic aryl group, e.g., phenyl) and a heteroaryl group (e.g., a mono-heteroaryl group, e.g., a 5-6 membered mono-heteroaryl group), which can be attached at the aryl ring or at the heteroaryl ring. The "aryl-heteroaryl" includes but is not limited to monocyclic aryl-heteroaryl. The term "9-12 membered aryl-heteroaryl" refers to an aryl-heteroaryl group containing a total of 9-12 ring atoms, e.g., benzene-5-6 membered nitrogen-containing mono-heteroaryl.

[0137] The term "heteroaryl-cycloalkyl" refers to a fused ring group of a heteroaryl group (e.g., a mono-heteroaryl group, e.g., a 5-6 membered mono-heteroaryl group) and a cycloalkyl group (e.g., a C 4-6 monocyclic cycloalkyl group), which can be attached at the heteroaryl ring or at the cycloalkyl group. The "heteroaryl-cycloalkyl" includes but is not limited to mono-heteroaryl-monocyclic cycloalkyl. The term "9-10 membered heteroaryl-cycloalkyl" refers to a heteroaryl-cycloalkyl group containing a total of 9-10 ring atoms, e.g., 4-6 membered nitrogen-containing mono-heteroaryl-C 4-6 monocyclic cycloalkyl.

[0138] ​The term "substituted" means that one or more (e.g., 1, 2, 3, or 4) hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency is not exceeded, and that the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0139] If a substituent is described as "optionally substituted" the substituent can be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with a selection of one or more of a list of substituents, then one or more hydrogens on the carbon (to the extent there are any hydrogens present) can each be replaced with an independently selected optional substituent. If a nitrogen of a substituent is described as being optionally substituted with a selection of one or more of a list of substituents, then one or more hydrogens on the nitrogen (to the extent there are any hydrogens present) can each be replaced with an independently selected optional substituent.

[0140] If a substituent is described as being "independently selected from" a group, each substituent is selected independently of the other(s). Thus, each substituent can be the same or different from the other (other) substituent(s).

[0141] As used herein, the term "one or more" means 1 or more than 1, e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, under reasonable conditions.

[0142] Unless indicated, as used herein, the point of attachment of a substituent can be from any suitable position of the substituent.

[0143] The present application also includes all pharmaceutically acceptable isotopically enriched compounds of the present application, which are identical to those recited herein, but for one or more atoms having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. Examples of isotopes suitable for inclusion in the compounds of the application include, but are not limited to, isotopes of hydrogen, such as 2 H, 3 H); isotopes of carbon, such as 11 C, 13 C and 14 C); isotopes of chlorine, such as 36 Cl); isotopes of fluorine, such as 18 F); isotopes of iodine, such as 123 I and 125 I); isotopes of nitrogen, such as 13 N and 15 N); isotopes of oxygen, such as 15 O, 17 O and 18 O); isotopes of phosphorus, such as32 P); and isotopes of sulfur (e.g., ... 35 S). The term "stable isotopic derivative" refers to a stable compound formed by replacing one or more atoms in the compound of the present invention with atoms having the same atomic number but different atomic mass or mass number from the dominant atomic mass or mass number in nature.

[0144] The term "stereoisomer" refers to isomers of a compound formed due to the presence of at least one asymmetric center. In compounds having one or more (e.g., 1, 2, 3, or 4) asymmetric centers, racemic mixtures, single enantiomers, diastereomer mixtures, and single diastereomers can occur. Specific individual molecules can also exist as geometric isomers (cis / trans). The compounds of this invention can exist as mixtures of two or more different structural forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. For example, nitroso-oximes can exist in equilibrium in solution in the following tautomer forms:

[0145]

[0146] It should be understood that the scope of this application covers all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%).

[0147] Unless otherwise specified, the compounds of the present invention are intended to exist as stereoisomers (including cis and trans isomers), optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, trans-blocking isomers, or mixtures thereof. The compounds of the present invention may exhibit more than one type of isomerism and may consist of mixtures thereof (e.g., racemic mixtures and diastereomer pairs).

[0148] This invention encompasses all possible crystalline forms or polymorphs of the compounds of this invention, which may be a single polymorph or a mixture of more than one polymorph in any proportion. It should also be understood that certain compounds of this invention may exist in a free form for therapeutic purposes, or, where appropriate, in their pharmaceutically acceptable derivative forms. In this invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, solvates, metabolites, or prodrugs that, upon administration to a patient in need, can directly or indirectly provide the compounds of this invention or their metabolites or residues. Therefore, when referring to "compounds of this invention" herein, it is also intended to cover the various derivative forms of the compounds described above.

[0149] Pharmaceutically acceptable salts of the compounds of the present application include both acid and base addition salts. For example, hydrochloride, hexafluorophosphate, meglumine salts, and the like. A review of suitable salts is available in "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, 2002).

[0150] "Pharmaceutically acceptable carrier" in the present application means a diluent, adjuvant, excipient or vehicle with which a therapeutic agent is administered, and which is suitable for use in contact with the tissue of humans and / or other animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio, as well as nontoxic to the recipients thereof.

[0151] Pharmaceutically acceptable carriers that can be employed in the pharmaceutical compositions of the application include, but are not limited to, sterile aqueous, nonaqueous, and mixed media broken down into excipients, adjuvants, vehicles, and agents, for example, water and oils, including those of both vegetable and animal origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water is an exemplary carrier when the pharmaceutical composition is administered intravenously. Saline and aqueous dextrose and glycerol can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, white

[0152] The pharmaceutical compositions of the present application can act systemically and / or topically. To this end, they can be administered by suitable routes, such as by injection, intravenously, intraarterially, subcutaneously, intraperitoneally, intramuscularly, intratumorally, or transdermally; or orally, buccally, nasally, transmucosally, topically, in the form of an ophthalmic preparation, or by inhalation.

[0153] For these administration routes, the pharmaceutical compositions of the present application can be administered in suitable dosage forms.

[0154] The dosage forms include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, syrups.

[0155] The term "effective amount" as used herein refers to an amount of a compound that, when administered, will relieve to some extent one or more of the symptoms of the disorder being treated.

[0156] Dosage regimens can be adjusted to provide the optimum desired response. For example, a single bolus can be administered, several divided doses can be administered over time or the dose can be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is noted that dose values can vary as to the type and severity of the condition being alleviated, and can include single or multiple doses. It is further understood that for any particular individual, specific dose regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions.

[0157] The amount of a compound of the present application that is administered can depend on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the judgment of the prescribing physician. In general, an effective dose is in the range of about 0.0001 to about 50 mg per kg body weight per day, e.g., about 0.01 to about 10 mg / kg / day (single or divided doses). For a 70-kg human, this amount would correspond to about 0.007 mg / day to about 3500 mg / day, e.g., about 0.7 mg / day to about 700 mg / day. In some instances, dosage levels less than the lower limit of the aforesaid range can be more than adequate, while in other cases still larger doses can be employed, or the stated larger doses can be administered as a fraction or in multiple doses over time.

[0158] The amount or quantity of a compound of the present application in a pharmaceutical composition can be about 0.01 mg to about 1000 mg, suitably 0.1-500 mg, preferably 0.5-300 mg.

[0159] The term "treating" as used herein, means reversing, alleviating, or ameliorating a disorder or condition to which such term applies, or one or more symptoms of such disorder or condition, or preventing such disorder or condition, or one or more symptoms of such disorder or condition, unless otherwise indicated.

[0160] "Individual" as used herein includes a human or non-human animal. Exemplary human individuals include a human individual (referred to as a patient) suffering from a disease (e.g., a disease described herein) or a normal individual. "Non-human animals" in the present application include all vertebrates, e.g., non-mammals (e.g., birds, amphibians, reptiles) and mammals, e.g., non-human primates, farm animals, and / or domestic animals (e.g., sheep, dog, cat, cow, pig, etc.).

[0161] The compounds of the present application can exist in solvate (preferably hydrate) form, wherein the compound of the present application contains a polar solvent, in particular, for example, water, methanol or ethanol, as an integral part of the crystal lattice of said compound. The amount of polar solvent, in particular, water, can be present in stoichiometric or non-stoichiometric amounts.

[0162] Also included within the scope of the present application are metabolites of compounds of the present application, i.e., substances formed by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, and the like, of the compounds of the present application when administered to an animal (e.g., human). The present application also includes compounds formed by a process comprising contacting a compound of the present application with a mammal for a period of time sufficient to yield a metabolic product of the compound.

[0163] The present application further includes within its scope prodrugs of the compounds of the present application, which are certain derivatives of the compounds of the present application that have less or no pharmacological activity themselves but are converted to the active compounds of the present application after administration to the body. Generally, such prodrugs will be functional derivatives of the compounds of the present application, which are readily converted to the active compounds of the present application in vivo. For examples of prodrugs, see "Pro-drugs as Novel Delivery Systems", Vol. 14 of the A.C.S. Symposium Series (T. Higuchi and V. Stella) and "Bioreversible Carriers in Drug Design," Pergamon Press, 1987 (ed. E. B. Roche, American Pharmaceutical Association). Prodrugs of the present application can be prepared using certain moieties known in the art as "pro-moieties" (e.g., as described in "Design of Prodrugs", H. Bundgaard (Elsevier, 1985)) to replace appropriate functionalities present in the compounds of the present application.

[0164] The present application also encompasses compounds of the present application that contain protecting groups. It can be necessary and / or desirable to protect sensitive or reactive groups m any of the molecules involved in synthesizing compounds of the present application, and / or to protect vulnerable sites of these molecules, by forming chemically protected forms of the compounds of the application at any stage during the preparation. This can be achieved by means of conventional protecting groups, such as those described in Protective Groups in Organic Chemistry, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which are incorporated herein by reference. The protecting groups can be removed at a suitable subsequent stage using methods known from the art.

[0165] As used herein represents a single or double bond.

[0166] As used herein represents a double bond position is not defined, but still guarantees that the ring in which it is located is aromatic.

[0167] As used herein represents the presence or absence of a bond in the structural formula.

[0168] As used herein "room temperature" means 15-30 °C.

[0169] Preparation method

[0170] Synthesis of compound V-A

[0171]

[0172] R 1 , R 2 , R 3 , R 4 , L are defined as in formula V above, but n + p + q in L is not 0, and in R 3 -L-COOH and R 3 -L-COCl, -L-R 3 is attached to the carbon atom to which -COOH or -COCl is attached, so that -L-R 3 is attached to the carbon atom to which -COOH or -COCl is attached, so that -L-R

[0173] Step 1: Nitration of compound V-A-1 to produce compound V-A-2.

[0174] In certain embodiments, the nitration reaction can be carried out using a nitrating agent such as concentrated nitric acid or fuming nitric acid. In certain embodiments, the reaction is carried out in a solvent, and a solvent such as acetic acid, propionic acid, or butyric acid can be used. In certain embodiments, the reaction temperature is from 60 °C to 150 °C.

[0175] Second step: Compound V-A-2 is chlorinated to form compound V-A-3.

[0176] In certain embodiments, the chlorination reaction can be carried out using a chlorinating agent such as POCI3, PCI3, or PCI5. In certain embodiments, the reaction is carried out in a solvent, and a solvent such as DCM or DCE can be used. In certain embodiments, the reaction is carried out under solvent-free conditions. In certain embodiments, the reaction temperature is from rt to 110 °C.

[0177] Third step: Compound V-A-3 is reacted with R 2 NH2under the action of a base to form compound V-A-4.

[0178] In certain embodiments, a base such as TEA or DIPEA can be used. In certain embodiments, the reaction is carried out in a solvent, and a solvent such as NMP or DMF can be used. In certain embodiments, the reaction temperature is from 0 °C to 153 °C.

[0179] Fourth step: Compound V-A-4 is reduced to form compound V-A-5.

[0180] In certain embodiments, a reducing agent such as sodium hydrosulfite, SnCl2, Fe, Zn, or Pd / C can be used. In certain embodiments, the reaction is carried out in a solvent, and a solvent such as MeOH, EtOH, THF, or 1,4-dioxane can be used. In certain embodiments, the reaction temperature is from rt to 110 °C.

[0181] Fifth step: Compound V-A-5 is condensed with R 3 -L-COOH under the action of a base to form compound V-A-6, or compound V-A-5 is directly reacted with R 3 -L-COCl under the action of a base to form compound V-A-6.

[0182] In certain embodiments, a condensing agent such as HATU, DCC / NHS, EDCI / HOBt, or HBTU can be used. In certain embodiments, a base such as TEA or DIPEA can be used. In certain embodiments, the reaction is carried out in a solvent, and a solvent such as THF, DCM, DCE, CH3CN, 1,4-dioxane, or DMF can be used. In certain embodiments, the reaction temperature is from 0 °C to 160 °C.

[0183] Step 6: Compound V-A-6 is subjected to ring closure under basic conditions to generate compound V-A-7.

[0184] In certain embodiments, the reaction can be carried out using a base such as TEA or DIPEA. In certain embodiments, the reaction is carried out in a solvent, which can be MeOH, EtOH, THF, CH3CN, or 1,4-dioxane, etc. In certain embodiments, the reaction temperature is from 60 °C to 100 °C.

[0185] Step 7: Compound V-A-7 is oxidized to generate compound V-A-8.

[0186] In certain embodiments, the oxidation reaction can be carried out using an oxidant such as m-CPBA, H2O2, or CH3COOOH. In certain embodiments, the reaction is carried out in a solvent, which can be DCM, DCE, CHCl3, or DMF, etc. In certain embodiments, the reaction temperature is from 0 °C to 160 °C.

[0187] Step 8: Compound V-A-8 is subjected to a reaction with R 4 MgX to generate compound V-A-9,

[0188] wherein X is selected from chlorine, bromine, iodine.

[0189] In certain embodiments, the reaction is carried out in a solvent, which can be selected from THF, 1,4-dioxane, toluene, etc. In certain embodiments, the reaction temperature is from -20 °C to 100 °C.

[0190] Step 9: Compound V-A-9 is subjected to a coupling reaction (e.g., Suzuki reaction, Stille reaction) with R 1 - boronic acid or R 1 - boronic ester or R 1 - organotin compound (e.g., R 1 Sn(n-Bu)3) to generate compound V-A.

[0191] In certain embodiments, the reaction can be carried out using a catalyst such as Pd(PPh3)4or Pd(dppf)Cl2·CH2Cl2. In certain embodiments, the reaction can be carried out using a base selected from Cs2CO3, K3PO4, Na2CO3, AcOK, NaHCO3, and K2CO3, etc. In certain embodiments, the reaction is carried out in a solvent, which can be selected from 1,4-dioxane / H2O, DMF / H2O, DMSO / H2O, CH3CN / H2O, and toluene / H2O, etc. In certain embodiments, the reaction temperature is from 60 °C to 180 °C.

[0192] Synthesis of compound VI-A

[0193]

[0194] R 1 , R 2 , R 3 , R 4 , L is as defined above in formula VI, but n + p + q in L is not 0, and in R 3 -L-COOH and R 3 -L-COCl, -L-R 3 is attached to the carbon atom of -COOH or -COCl, so that -L-R 3 is attached to the carbon atom of the imidazole ring.

[0195] Step 1: Compound VI-A-1 is subjected to nitration to form compound VI-A-2.

[0196] In certain embodiments, the nitration reaction can be carried out using a nitration agent selected from concentrated nitric acid, fuming nitric acid, and the like. In certain embodiments, the reaction is carried out in a solvent, and a solvent that can be used is acetic acid, propionic acid, or butyric acid, and the like. In certain embodiments, the reaction temperature is from 60 °C to 150 °C.

[0197] Step 2: Compound VI-A-2 is subjected to chlorination to form compound VI-A-3.

[0198] In certain embodiments, the chlorination reaction can be carried out using a chlorinating agent selected from POCI3, PCI3, or PCI5, and the like. In certain embodiments, the reaction is carried out in a solvent, and a solvent that can be used is DCM or DCE, and the like. In certain embodiments, the reaction is carried out in a solvent-free condition. In certain embodiments, the reaction temperature is from rt to 110 °C.

[0199] Step 3: Compound VI-A-3 is subjected to substitution with R 2 NH2 in the presence of a base to form compound VI-A-4.

[0200] In certain embodiments, the base that can be used is selected from TEA, DIPEA, and the like. In certain embodiments, the reaction is carried out in a solvent, and a solvent that can be used is selected from NMP, DMF, and the like. In certain embodiments, the reaction temperature is from 0 °C to 153 °C.

[0201] Step 4: Compound VI-A-4 is reduced to form compound VI-A-5.

[0202] In certain embodiments, the reduction reaction can be carried out using a reducing agent such as sodium hydrosulfite, SnCl2, Fe, Zn, or Pd / C. In certain embodiments, the reaction is carried out in a solvent, and the solvent that can be used is MeOH, EtOH, THF, or 1,4-dioxane. In certain embodiments, the reaction temperature is from rt to 110 °C.

[0203] Step 5: Compound VI-A-5 is condensed with a base under the action of a condensing agent to generate compound VI-A-6, or compound VI-A-5 is directly condensed with R 3 -L-COOH to generate compound VI-A-6, or compound VI-A-5 is directly condensed with R 3 -L-COCl to generate compound VI-A-6.

[0204] In certain embodiments, the condensing agent that can be used is HATU, DCC / NHS, EDCI / HOBt, or HBTU. In certain embodiments, the base that can be used is TEA or DIPEA. In certain embodiments, the reaction is carried out in a solvent, and the solvent that can be used is THF, DCM, DCE, CH3CN, 1,4-dioxane, or DMF. In certain embodiments, the reaction temperature is from 0 °C to 160 °C.

[0205] Step 6: Compound VI-A-6 is ring-closed under basic conditions to generate compound VI-A-7.

[0206] In certain embodiments, the reaction can be carried out using a base such as TEA or DIPEA. In certain embodiments, the reaction is carried out in a solvent, and the solvent that can be used is MeOH, EtOH, THF, CH3CN, or 1,4-dioxane. In certain embodiments, the reaction temperature is from 60 °C to 100 °C.

[0207] Step 7: Compound VI-A-7 is oxidized to generate compound VI-A-8.

[0208] In certain embodiments, the oxidation reaction can be carried out using an oxidizing agent such as m-CPBA, H2O2, or CH3COOOH. In certain embodiments, the reaction is carried out in a solvent, and the solvent that can be used is DCM, DCE, CHCl3, or DMF. In certain embodiments, the reaction temperature is from 0 °C to 160 °C.

[0209] Step 8: Compound VI-A-8 is reacted with R 4 MgX to generate compound VI-A-9,

[0210] wherein X is selected from chlorine, bromine, iodine.

[0211] In certain embodiments, the reaction is carried out in a solvent, which can be selected from THF, 1,4-dioxane, toluene, and the like. In certain embodiments, the reaction temperature is from -20 °C to 100 °C.

[0212] Step 9: Compound VI-A-9 is subjected to bromination to generate compound VI-A-10.

[0213] In certain embodiments, the reaction can be carried out using a brominating agent selected from bromine, NBS, and dibromohydantoin, and the like. In certain embodiments, the reaction is carried out in a solvent, which can be selected from acetic acid, 1,4-dioxane, DMF, EA, and the like. In certain embodiments, the reaction temperature is from -20 °C to 100 °C.

[0214] Step 10: Compound VI-A-10 is subjected to coupling reaction (for example, Suzuki reaction, Stille reaction) with R 1 - boronic acid or R 1 - boronic ester or R 1 - organotin compound (for example, R 1 Sn(n-Bu)3) to generate compound VI-A.

[0215] In certain embodiments, the reaction can be carried out using a catalyst selected from Pd(PPh3)4and Pd(dppf)Cl2·CH2Cl2, and the like. In certain embodiments, the reaction can be carried out using a base selected from Cs2CO3, K3PO4, Na2CO3, AcOK, NaHCO3, and K2CO3, and the like. In certain embodiments, the reaction is carried out in a solvent, which can be selected from 1,4-dioxane / H2O, DMF / H2O, DMSO / H2O, CH3CN / H2O, toluene / H2O, and the like. In certain embodiments, the reaction temperature is from 60 °C to 180 °C.

[0216] Advantages

[0217] The compounds of the present application have obvious agonistic activity on NLRP3 and its signaling pathway, have no obvious toxic side effects, and can be used for the treatment of diseases (for example, tumors) related to NLRP3 inflammasome activity. Examples

[0218] The present application is further described below in conjunction with examples, which are provided not intended to limit the scope of the present application.

[0219] The abbreviations in the present application have the following meanings:

[0220]

[0221]

[0222] The structure of the compound was confirmed by nuclear magnetic resonance spectroscopy (H NMR) or mass spectrometry (MS). 1 H NMR) or mass spectrometry (MS).

[0223] The reaction was monitored by thin layer chromatography (TLC) or LC-MS.

[0224] 1 H NMR spectroscopy: Bruker superconducting nuclear magnetic resonance spectrometer (model AVACE III HD 400MHz). Thin layer chromatography was performed using silica gel GF 254 as the stationary phase.

[0225] LC / MS mass spectrometer: Aglient 1260 Infinity / Aglient 6120 Quadrupole

[0226] The compounds of the present application can be separated and purified by chromatography on silica gel plates, column chromatography on silica gel, preparative high performance liquid chromatography (Prep-HPLC), flash column chromatography.

[0227] Prep-HPLC used Agilent 1260 preparative liquid chromatography, detection wavelength: 214nm, 254nm; column: Waters SunFire Prep C18 OBD (19mm x 150mm x 5.0μm); column temperature: 25℃.

[0228] Elution conditions:

[0229] Condition 1: 10%-90% acetonitrile and 90%-10% ammonium bicarbonate aqueous solution (0.05%); 0-16min; flow rate: 25mL / min;

[0230] Condition 2: 30%-90% acetonitrile and 70%-10% ammonium bicarbonate aqueous solution (0.05%); 0-16min; flow rate: 25mL / min;

[0231] Condition 3: 10%-90% acetonitrile and 90%-10% ammonium bicarbonate aqueous solution (0.05%); 0-16min; flow rate: 20mL / min;

[0232] Column chromatography generally used 200-300 mesh silica gel (Qingdao Marine) as the stationary phase. Eluent system A: dichloromethane and methanol; eluent system B: petroleum ether and ethyl acetate, the volume ratio of the solvents was adjusted according to the different polarity of the compounds.

[0233] Flash column chromatography used Biotage flash column chromatograph.

[0234] Microwave reaction used Biotage Initiator+microwave reactor.

[0235] In the following examples, the reaction temperature is room temperature (15-30°C) unless otherwise specified.

[0236] The reagents used in this application were purchased from Acros Organics, Aldrich Chemical Company or TCI Chemicals, etc.

[0237] Example 1: Synthesis of 2-(3-(benzyloxy)propyl)-l,4-dimethyl-7-(lH-pyrazol-3-yl)-lH- imidazo[4,5-d]thieno[3,2-b]pyridine (1)

[0238] 3-(l,4-dimethyl-7-(lH-pyrazol-3-yl)-lH-imidazo[4,5-d]thieno[3,2-b]pyridin-2-yl)propanol (2)

[0239]

[0240] First Step: Synthesis of compound (4-benzyloxy)-(7-(methylamino)thieno[3,2-b]pyridin-6- yl)butanamide (1b)

[0241] DIPEA (1.44 g, 11.16 mmol), HATU (1.06 g, 2.79 mmol) were added to 4-benzyloxybutanoic acid (541.80 mg, 2.79 mmol), compound la (500 mg, 2.79 mmol) in DMF (4 mL) sequentially, then the reaction was stirred at room temperature for 5 hr. The reaction was quenched by water, extracted by dichloromethane. The organic layer was dried and concentrated to give compound lb (850 mg).

[0242] MS (ESI, m / z): 356.1 [M+H] +

[0243] Second Step: Synthesis of compound 2-(3-(benzyloxy)propyl)-l-methyl-lH-imidazo[4,5-d]thieno[3,2-b]pyridine (1c)

[0244] Sodium hydroxide (184.55 mg, 4.61 mmol) was added to compound lb (820 mg, 2.31 mmol) in ethanol (4 mL). Then the reaction was heated to 90 °C for 2 hr. The reaction was concentrated under reduced pressure, extracted by water and ethyl acetate. The organic layer was dried and concentrated, then purified by column chromatography (eluent system A) to give compound lc (500 mg).

[0245] MS (ESI, m / z): 338.1 [M+H] +

[0246] Step 3: Synthesis of compound 2-(3-(benzyloxy)propyl)-l-methyl-lH-imidazo[4,5- d]thieno[3,2-b]pyridine-2,5-dioxide (Id)

[0247] Compound lc (500 mg, 1.48 mmol) was dissolved in DCM (5 mL) and m-chloroperbenzoic acid (382.29 mg, 2.22 mmol) was added portionwise. The reaction was stirred at room temperature for 4 hr. The reaction was quenched with saturated sodium bicarbonate solution and extracted with dichloromethane. The organic layer was dried and concentrated. The residue was purified by column chromatography (eluent system A) to give compound Id (400 mg). MS (ESI, m / z): 354.1 [M+H] +

[0248] Step 4: Synthesis of compound 2-(3-(benzyloxy)propyl)-l,4-dimethyl-lH-imidazo[4,5- d]thieno[3,2-b]pyridine (le)

[0249] Compound Id (500 mg, 1.41 mmol) was dissolved in THF (4 mL) and methyl magnesium bromide (2.5 M, 5.66 mL) was added slowly. The reaction was stirred at room temperature for 1 hr. The reaction was quenched with water and extracted with ethyl acetate. The organic layer was dried and concentrated. The residue was purified by column chromatography (eluent system B) to give compound le (400 mg). MS (ESI, m / z): 352.1 [M+H] + .

[0250] Step 5: Synthesis of compound 2-3(benzyloxy)propyl)-7-bromo-l,4-dimethyl-lH- imidazo[4,5-d]thieno[3,2-b]pyridine (If)

[0251] Compound le (400 mg, 1.14 mmol) was dissolved in acetic acid (1 mL) and chloroform (1 mL) and NBS (405.12 mg, 2.28 mmol) was added portionwise. The reaction was stirred at room temperature for 2 hr. The reaction was extracted with dichloromethane and water. The organic layer was dried and concentrated. The residue was purified by column chromatography (eluent system B) to give compound If (300 mg). MS (ESI, m / z): 430.1 [M+H] +1 H NMR (400 MHz, MeOD) δ 7.49 (s, 1H), 7.16 - 7.07 (m, 5H), 4.40 (s, 2H), 3.85 (s, 3H), 3.64 (t, J = 5.8 Hz, 2H), 3.08 (t, J = 7.4 Hz, 2H), 2.79 (s, 3H), 2.22 - 2.11 (m, 2H).

[0252] Step 6: Synthesis of compound 2-(3-(benzyloxy)propyl)-l,4-dimethyl-7-(lH- pyrazol-3-yl)-lH-imidazo[4,5-d]thieno[3,2-b]pyridine (1)

[0253] Pd(dppf)Cl2(101.91 mg, 139.42 μmol), cesium carbonate (181.24 mg, 557.67 μmol) were added to compound Ik (64.93 mg, 334.60 μmol), compound If (120 mg, 278.84 μmol) in water / dioxane. The reaction was heated to 100 °C for 16 hr under N2protection. After concentration, compound 1 (50 mg) was obtained by column chromatography separation and purification (eluent system A). MS (ESI, m / z): 418.1 [M+H] +

[0254] 1 H NMR (400 MHz, DMSO-d6) δ 13.07 (s, 1H), 7.86 (d, J = 9.3 Hz, 2H), 7.35 - 7.22 (m, 5H), 6.87 (s, 1H), 4.50 (s, 2H), 3.97 (d, J = 8.1 Hz, 3H), 3.59 (t, J = 6.2 Hz, 2H), 3.01 (t, J = 7.5 Hz, 2H), 2.74 (s, 3H), 2.16 - 2.05 (m, 2H).

[0255] Step 7: Synthesis of compound 3-(l,4-dimethyl-7-(lH-pyrazol-3-yl)-lH- imidazo[4,5-d]thieno[3,2-b]pyridin-2-yl)propan-l-ol (2)

[0256] Bromotrimethylsilane (47.90 mg, 191.60 μmol) was slowly added to compound 1 (40 mg, 95.80 μmol) in DCM (5 mL) at room temperature. After stirring for five minutes at room temperature, the reaction was diluted with ethyl acetate and water, adjusted to pH = 8, and extracted with ethyl acetate. After drying and concentration of the organic layer, compound 2 (3 mg) was obtained by Prep-HPLC separation and purification (elution condition 1).

[0257] MS (ESI, m / z): 328.1 [M+H] +

[0258] 1H NMR (400 MHz, DMSO-d6) δ 13.09 (s, 1H), 7.84 (s, 2H), 6.85 (d, J = 1.8 Hz, 1H), 4.66 (s, 1H), 3.97 (s, 3H), 3.55 (d, J = 4.3 Hz, 2H), 2.97 (t, J = 7.6 Hz, 2H), 2.74 (s, 3H), 1.97 (dd, J = 14.3, 6.7 Hz, 2H).

[0259] Example 2: 1,4-Dimethyl-2-(3-(4-methylpiperazin-1-yl)propyl)-7-(1H-pyrazol-3-yl)- 1H-imidazo[4,5-d]thieno[3,2-b]pyridine (3)

[0260] 1,4-Dimethyl-2-(3-(4-methylpiperazin-1-yl)propyl)-7-(1H-pyrazol-3-yl)-1H- imidazo[4,5-d]thieno[3,2-b]pyridine hydrochloride (3a)

[0261]

[0262] First Step: 3-(7-Bromo-1,4-dimethyl-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-2-yl)propyl- 1-ol (2a)

[0263] Compound 1f (320 mg, 0.74 mmol) was added into 3 mL trifluoroacetic acid, heated to 80 °C and stirred for 16 h. The reaction was concentrated, re-dissolved in 3 mL MeOH, adjusted to pH = 9.0 with 2 N sodium hydroxide solution, and stirred for another 30 min. Compound 2a (180 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 340.0 [M+H] + .

[0264] Second Step: 7-Bromo-2-(3-chloropropan-1,4-dimethyl-1H-imidazo[4,5-d]thieno[3,2- b]pyridine (2b)

[0265] Compound 2a (180 mg, 0.53 mmol), DMF (4 mg, 0.05 mmol) was added into 4 mL dichlorosulfoxide / tetrahydrofuran (1:1), and stirred at room temperature for 12 h. After the reaction solvent was rotary evaporated, compound 2b (150 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 358.0 [M+H] + .

[0266] Step 3: 7-bromo-1,4-dimethyl-2-(3-(4-methylpiperazin-1 -yl)propyl)-1 H- imidazo[4,5-d]thieno[3,2-b]pyridine (2c)

[0267] Compound 2b (150 mg, 0.42 mmol), N-methylpiperazine (9 mg, 0.84 mmol), tetrabutylammonium iodide (29 mg, 0.08 mmol), DIPEA (109 mg, 0.84 mmol) were added into 5 mL of toluene and heated to 100 °C for 12 h under N2. The reaction solvent was removed by rotary evaporation and the residue was purified by flash column chromatography (eluent system A) to give compound 2c (140 mg). MS (ESI, m / z): 422.1 [M+H] + .

[0268] Step 4: 1,4-dimethyl-2-(3-(4-methylpiperazin-1 -yl)propyl)-7-(1 H-pyrazol-3-yl)- 1 H-imidazo[4,5-d]thieno[3,2-b]pyridine (3)

[0269] Compound 2c (140 mg, 0.33 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 1 H-pyrazole (128 mg, 0.66 mmol), Pd(dppf)Cl2(57 mg, 0.07 mmol), sodium carbonate (70 mg, 0.66 mmol) were added into 5 mL of DMF and 1 mL of water and heated to 110 °C for 3 h under N2. The reaction solvent was removed by rotary evaporation and the residue was purified by Prep-HPLC (elution condition 2) to give compound 3. MS (ESI, m / z): 410.2 [M+H] +

[0270] Step 5: 1,4-dimethyl-2-(3-(4-methylpiperazin-1 -yl)propyl)-7-(1 H-pyrazol-3-yl)- 1 H-imidazo[4,5-d]thieno[3,2-b]pyridine hydrochloride (3a)

[0271] To the collected fraction of Prep-HPLC purification of compound 3 from step 4, 0.5 mL of 6 N hydrochloric acid was added, followed by lyophilization to give compound 3a (24 mg).

[0272] MS (ESI, m / z): 410.2 [M+H] + .

[0273] 1H NMR (DMSO-d6, 400 MHz) δ 13.07 (s, 1H), 11.93-11.82 (m, 2H), 7.93-7.82 (m, 2H), 6.86 (s, 1H), 3.99 (s, 3H), 2.94 (t, J = 7.4 Hz, 2H), 2.74 (s, 3H), 2.51-2.24 (m, 10H), 2.07 (s, 3H), 2.05-1.92 (m, 2H).

[0274] Example 3: 2-(3-(azetidin-1-yl)propyl)-1,4-dimethyl-7-(1H-pyrazol-3-yl)-1H- imidazo[4,5-d]thieno[3,2-b]pyridine (5)

[0275]

[0276] First Step: 2-(3-(azetidin-1-yl)propyl)-7-bromo-1,4-dimethyl-1H-imidazo[4,5- d]thieno[3,2-b]pyridine (3a)

[0277] Compound 2b (50 mg, 0.14 mmol), azetidine (48 mg, 0.84 mmol), tetrabutylammonium iodide (10 mg, 0.03 mmol), TEA (85 mg, 0.84 mmol) were added into 3 mL of toluene, heated to 100 °C for 16 h under N2protection. The reaction solvent was rotary evaporated to dryness to give compound 3a (53 mg), which was used directly for the next step. MS (ESI, m / z): 379.1 [M+H] + .

[0278] Second Step: 2-(3-(azetidin-1-yl)propyl)-1,4-dimethyl-7-(1H-pyrazol-3-yl)-1H- imidazo[4,5-d]thieno[3,2-b]pyridine (5)

[0279] Compound 3a (53 mg, 0.13 mmol), 1H-pyrazole-3-boronic acid pinacol ester (61 mg, 0.31 mmol), Pd(dppf)Cl2(10 mg, 0.01 mmol), sodium carbonate (40 mg, 0.38 mmol) were added into 4 mL of DMF and 1 mL of water mixed solvent, heated to 100 °C for 4 h under N2protection. The reaction solvent was concentrated to dryness under reduced pressure, separated and purified by Prep-HPLC (elution condition 3), and lyophilized to give compound 5 (16 mg)

[0280] MS (ESI, m / z): 367.2 [M+H] + .

[0281] MS (ESI, m / z): 367.2 [M+H]1 H NMR (DMSO-d6, 400 MHz) δ 13.09 (s, 1H), 7.85 (d, J = 3.4 Hz, 2H), 6.85 (d, J = 2.3 Hz, 1H), 4.00-3.96 (m, 3H), 3.04 (m, 4H), 2.92 (t, J = 7.6 Hz, 2H), 2.74 (s, 3H), 2.44 (t, J = 6.8 Hz, 2H), 1.98-1.90 (m, 2H), 1.84-1.72 (m, 2H).

[0282] Example 4: 3-(4-isopropyl-l-methyl-7-(lH-pyrazol-3-yl)-lH-imidazo[4,5- d]thieno[3,2-b]pyridin-2-yl)propan-l-ol (6)

[0283]

[0284] First Step: 2-(3-(benzyloxy)propyl)-4-isopropyl-l-methyl-lH-imidazo[4,5- d]thieno[3,2-b]pyridine (4a)

[0285] Compound Id (400 mg, 1.13 mmol), isopropyl magnesium bromide (830 mg, 5.65 mmol) were added into 8 mL of anhydrous tetrahydrofuran, and the reaction was stirred at 25 °C for 8 h. The reaction was poured into ice water, and the organic phase was concentrated after extraction with dichloromethane, and then purified by flash column chromatography (eluent system A) to obtain compound 4a (310 mg). MS (ESI, m / z): 380.2 [M+H] + .

[0286] Second Step: 2-(3-(benzyloxy)propyl)-7-bromo-4-isopropyl-l-methyl-lH- imidazo[4,5-d]thieno[3,2-b]pyridine (4b)

[0287] Compound 4a (310 mg, 0.82 mmol), N-bromosuccinimide (292 mg, 1.64 mmol) were added into 6 mL of a mixed solvent of chloroform / ice acetic acid (1:1), and the reaction was stirred at room temperature for 2 h. The reaction was poured into ice water, and the organic phase was concentrated after extraction with dichloromethane, and then purified by flash column chromatography (eluent system A) to obtain compound 4b (290 mg). MS (ESI, m / z): 458.1 [M+H] + .

[0288] Third Step: 3-(7-bromo-4-isopropyl-l-methyl-lH-imidazo[4,5-d]thieno[3,2- b]pyridin-2-yl)propan-l-ol (4c)

[0289] Compound 4b (290 mg, 0.63 mmol) was added into 3 mL trifluoroacetic acid, heated to 80 °C and stirred for 16 h. The reaction was concentrated to dryness, re-dissolved in 3 mL MeOH, adjusted to pH = 9.0 with 2 N sodium hydroxide solution, and stirred for another 30 min. Compound 4c (150 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 368.1 [M+H] + .

[0290] Fourth step: 3-(4-isopropyl-l-methyl-7-(lH-pyrazol-3-yl)-lH-imidazo[4,5- d]thieno[3,2-b]pyridin-2-yl)propan-l-ol (6)

[0291] Compound 4c (31 mg, 0.085 mmol), 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrazole (33 mg, 0.17 mmol), Pd(dppf)Cl2(8 mg, 0.01 mmol), sodium carbonate (18 mg, 0.17 mmol) were added into 2.5 mL DMF and 0.5 mL water mixed solvent, heated to 110 °C and stirred for 3 h under N2protection. The reaction solvent was concentrated to dryness under reduced pressure, and compound 6 (5 mg) was separated and purified by Prep-HPLC (elution condition 1).

[0292] MS (ESI, m / z): 356.1 [M+H] + .

[0293] 1 H NMR (DMSO-d6, 400 MHz) δ 13.05 (s, 1H), 7.94 (s, 1H), 7.86 (s, 1H), 6.88 (s, 1H), 4.65 (t, J = 5.2 Hz, 1H), 3.99 (s, 3H), 3.88-3.74 (m, 1H), 3.65-3.52 (m, 2H), 2.98 (t, J = 7.6 Hz, 2H), 2.03-1.88 (m, 2H), 1.37 (d, J = 6.9 Hz, 6H).

[0294] Example 5: 4-(3-(l,4-dimethyl-7-(lH-pyrazol-3-yl)-lH-imidazo[4,5-d]thieno[3,2- b]pyridin-2-yl)propyl)-2,2-dimethylmorpholine (7)

[0295]

[0296] First Step: 4-(3-(7-bromo-1,4-dimethyl-1 H-imidazo[4,5-d]thieno[3,2-b]pyridin-2- yl)propyl)-2,2-dimethylmorpholine (5b)

[0297] Compound 2b (40 mg, 111.52 μmol), compound 5a (12.84 mg, 111.52 μmol), TEA (11.28 mg, 111.52 μmol), tetrabutylammonium iodide (41.19 mg, 111.52 μmol) were added into toluene (2 mL) at room temperature, and the mixture was heated to 99 °C for 16 h. The mixture was filtered by adding ethyl acetate, and the filtrate was concentrated to dryness. The residue was purified by Prep-HPLC (elution system A) to give compound 5b (35 mg). MS (ESI, m / z): 439.1 [M+H] +

[0298] Second Step: 4-(3-(1,4-dimethyl-7-(1 H-pyrazol-3-yl)-1 H-imidazo[4,5-d]thieno[3,2- b]pyridin-2-yl)propyl)-2,2-dimethylmorpholine (7)

[0299] Pd(dppf)Cl2(2.92 mg, 4.00 μmol), potassium carbonate (22.09 mg, 160.04 μmol) were added into a mixture of compound 5b (15.53 mg, 80.02 μmol), 5c (35 mg, 80.02 μmol) in dioxane (2 mL) / water (0.5 mL) at room temperature. The mixture was heated to 90 °C for 3 hr under N2. The mixture was diluted with ethyl acetate, filtered, and concentrated to give a crude product. The crude product was purified by Prep-HPLC (elution condition 1) to give compound 7 (5 mg).

[0300] MS (ESI, m / z): 425.2 [M+H] +

[0301] 1 H NMR (DMSO-d6, 400 MHz) δ 13.07 (s, 1H), 7.86 (d, J = 7.9 Hz, 2H), 6.86 (s, 1H), 3.99 (s, 3H), 3.58-3.52 (m, 2H), 2.97 (t, J = 7.5 Hz, 2H), 2.74 (s, 3H), 2.36 (t, J = 6.8 Hz, 2H), 2.29 (s, 2H), 2.16 (s, 2H), 2.00-1.93 (m, 2H), 1.14 (s, 6H).

[0302] Example 6: 3-(1,4-Dimethyl-7-(1H-pyrazol-3-yl)-1H-imidazo[4,5-c]quinolin-2- yl)propan-1-ol (8)

[0303]

[0304] First Step: 2-(3-Benzyloxy)propyl-7-bromo-1-methyl-1H-imidazo[4,5-c]quinoline 5- oxide (6b)

[0305] Compound 6a (607 mg, 1.48 mmol), mCPBA (385 mg, 2.23 mmol) were added into 10 mL dichloromethane, the reaction was stirred at room temperature for 4 h, quenched by ice water, extracted by dichloromethane, the organic phase was concentrated, then purified by flash column chromatography (eluent system A) to give compound 6b (494 mg). MS (ESI, m / z): 428.1 [M+H] + .

[0306] Second Step: (2-3 (benzyloxy) propyl)-7-bromo-1,4-dimethyl-1H-imidazo[4,5-c]quinoline (6c)

[0307] Compound 6b (494 mg, 1.16 mmol) was added into 8 mL anhydrous tetrahydrofuran, then methyl magnesium bromide (691 mg, 5.81 mmol) was added, the reaction was stirred at 25 °C for 1 h, quenched by ice water, extracted by dichloromethane, the organic phase was concentrated, then purified by flash column chromatography (eluent system A) to give compound 6c (140 mg). MS (ESI, m / z): 426.1 [M+H] + .

[0308] Third Step: 2-(3-(Methyloxy)propyl)-1,4-dimethyl-7-(1H-pyrazol-3-yl)-1H-imidazo[4,5- c]quinoline (6d)

[0309] Compound 6c (140 mg, 0.33 mmol), (1H-pyrazol-3-yl)borate (128 mg, 0.66 mmol), Pd(dppf)Cl2(57 mg, 0.07 mmol), sodium carbonate (56 mg, 0.66 mmol) were added into 5 mL DMF and 1 mL water mixed solvent, the reaction was stirred at 110 °C under N2 protection for 6 h. Filtered by diatomite, the filtrate was concentrated to dryness under reduced pressure, then purified by flash column chromatography (eluent system A) to give compound 6d (45 mg). MS (ESI, m / z): 412.2 [M+H] + .

[0310] Step 4: 3-(l,4-dimethyl-7-(lH-pyrazol-3-yl)-lH-imidazo[4,5-c]quinolin-2- yl)propan-l-ol (8)

[0311] Compound 6d (45 mg, 0.33 mmol) was added to 5 mL of trifluoroacetic acid, stirred at 65 °C for 10 hr, concentrated under reduced pressure, redissolved with methanol, pH was adjusted to 8-9 with saturated sodium bicarbonate, filtered, the filtrate was concentrated, and compound 8 (15 mg) was isolated by Prep-HPLC purification (elution condition 2).

[0312] MS (ESI, m / z): 322.2 [M+H] + .

[0313] 1 H NMR (DMSO-d6, 400 MHz) δ 13.09 (s, 1H), 8.50-8.59 (m, 2H), 8.07-8.14 (m, 1H), 7.86-7.87 (m, 1H), 6.95 (s, 1H), 4.66-4.67 (m, 1H), 4.24 (s, 3H), 3.59-3.60 (m, 2H), 3.11-3.13 (m, 2H), 2.91 (s, 3H), 1.95-1.98 (m, 2H).

[0314] Example 7: l,4-dimethyl-7-(lH-pyrazol-3-yl)-2-(3-(pyrrolidin-l-yl)propyl)-lH- imidazo[4,5-d]thieno[3,2-b]pyridine (9)

[0315]

[0316] Step 1: 7-bromo-l,4-dimethyl-2-(3-(pyrrolidin-l-yl)propyl)-lH-imidazo[4,5- d]thieno[3,2-b]pyridine (7a)

[0317] Compound 2b (30 mg, 0.08 mmol), tetrahydropyrrole (34 mg, 0.48 mmol), tetrabutylammonium iodide (15 mg, 0.04 mmol), TEA (48 mg, 0.48 mmol) were added to 5 mL of toluene, heated to 100 °C for 12 h under N2protection. The solvent was rotary evaporated and compound 7a (20 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 393.1 [M+H] + .

[0318] Second Step: 1,4-Dimethyl-7-(1H-pyrazol-3-yl)-2-(3-(pyrrolidin-1-yl)propyl)-1H- imidazo[4,5-d]thieno[3,2-b]pyridine (9)

[0319] Compound 7a (20 mg, 0.05 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H- pyrazole (20 mg, 0.10 mmol), Pd(dppf)Cl2(8 mg, 0.01 mmol), sodium carbonate (70 mg, 0.66 mmol) were added in 2.5 mL DMF and 0.5 mL water mixed solvent, heated to 110 °C under N2protection and stirred for 3 h. Concentrated under reduced pressure, separated and purified by Prep-HPLC (elution condition 2) to obtain compound 9 (6 mg)

[0320] MS (ESI, m / z): 381.2 [M+H] + .

[0321] 1 H NMR (DMSO-d6, 400 MHz) δ 13.08 (s, 1H), 7.93-7.82 (m, 2H), 6.86 (s, 1H), 3.99 (s, 3H), 2.99 (t, J = 7.4 Hz, 2H), 2.80-2.64 (m, 9H), 2.13-1.96 (m, 2H), 1.74 (s, 4H).

[0322] Example 8: 4-Isopropyl-1-methyl-2-(3-(4-methylpiperazin-1-yl)propyl)-7-(1H-pyrazol-3-yl)- 1H-imidazo[4,5-d]thieno[3,2-b]pyridine (10)

[0323]

[0324] First Step: 7-Bromo-2-(3-chloropropane)-4-isopropyl-1-methyl-1H-imidazo[4,5-d]thieno[3,2- b]pyridine (8a)

[0325] Compound 4c (150 mg, 0.41 mmol), DMF (3 mg, 0.04 mmol) were added in 4 mL dichlorosulfoxide / tetrahydrofuran (1:1) mixed solvent, stirred at room temperature for 12 h. After spinning dry the solvent, compound 8a (125 mg) was obtained by flash column chromatography (eluent system A).

[0326] MS (ESI, m / z): 386.0 [M+H] + .

[0327] Second Step: 7-bromo-4-isopropyl-l-methyl-2-(3-(4-methylpiperazin-l- yl)propyl)-lH-imidazo[4,5-d]thieno[3,2-b]pyridine (8b)

[0328] Compound 8a (125 mg, 0.32 mmol), N-methylpiperazine (20 mg, 1.92 mmol), tetrabutylammonium iodide (22 mg, 0.06 mmol), TEA (194 mg, 1.92 mmol) were added into 5 mL toluene, heated to 100 °C for 12 h under N2protection. The solvent was removed by rotary evaporation and the residue was purified by flash column chromatography (eluent system A) to give compound 8b (130 mg). MS (ESI, m / z): 450.1 [M+H] + .

[0329] Third Step: 4-isopropyl-l-methyl-2-(3-(4-methylpiperazin-l-yl)propyl)-7-(lH- pyrazol-3-yl)-lH-imidazo[4,5-d]thieno[3,2-b]pyridine (10)

[0330] Compound 8b (75 mg, 0.17 mmol), 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrazole (66 mg, 0.34 mmol), Pd(dppf)Cl2(16 mg, 0.02 mmol), sodium carbonate (36 mg, 0.34 mmol) were added into 5 mL of mixed solvent of DMF and 1 mL of water, heated to 110 °C for 3 h under N2protection. The solvent was removed by rotary evaporation and the residue was purified by Prep-HPLC (elution condition 2) to give compound 10 (20 mg).

[0331] MS (ESI, m / z): 438.2 [M+H] + .

[0332] 1 H NMR (DMSO-d6, 400 MHz) δ 13.05 (s, 1H), 7.93 (s, 1H), 7.86 (s, 1H), 6.88 (s, 1H), 3.98 (m, 3H), 3.86-3.74 (m, 1H), 2.94 (t, J = 7.3 Hz, 2H), 2.45-1.86 (m, 15H), 1.36 (d, J = 6.9 Hz, 6H).

[0333] Example 9: 4-(3-(l,4-dimethyl-7-(lH-pyrazol-3-yl)-lH-imidazo[4,5-d]thieno[3,2- b]pyridin-2-yl)propyl)morpholine (11)

[0334]

[0335] First Step: 4-(3-(7-bromo-1,4-dimethyl-1 H-imidazo[4,5-d]thieno[3,2-b]pyridin-2- yl)propyl)morpholine (9a)

[0336] Compound 2b (150 mg, 0.42 mmol), morpholine (7 mg, 0.84 mmol), tetrabutylammonium iodide (29 mg, 0.08 mmol), DIPEA (109 mg, 0.84 mmol) were added into 5 mL of toluene, heated to 100 °C for 12 h under N2protection. The solvent was removed by rotary evaporation and the residue was purified by flash column chromatography (eluent system A) to give compound 9a (140 mg). MS (ESI, m / z): 409.1 [M+H] + .

[0337] Second Step: 4-(3-(1,4-dimethyl-7-(1 H-pyrazol-3-yl)-1 H-imidazo[4,5-d]thieno[3,2- b]pyridin-2-yl)propyl)morpholine (11)

[0338] Compound 9a (140 mg, 0.34 mmol), (1 H-pyrazol-3-yl)boronic acid (77 mg, 0.68 mmol), Pd(dppf)Cl2(57 mg, 0.07 mmol), sodium carbonate (58 mg, 0.66 mmol) were added into 5 mL of DMF and 1 mL of water, heated to 110 °C for 3 h under N2protection. The solvent was concentrated under reduced pressure and the residue was purified by Prep-HPLC (elution condition 1) and lyophilized to give compound 11 (10 mg).

[0339] MS (ESI, m / z): 397.1 [M+H] + .

[0340] 1 H NMR (DMSO-d6, 400 MHz) δ 13.09 (s, 1H), 7.88-7.84 (m, 2H), 6.86 (s, 1H), 3.98 (s, 3H), 3.53 (m, 4H), 2.95 (t, J = 7.4 Hz, 2H), 2.75 (s, 3H), 2.49-2.31 (m, 6H), 1.99 (t, J = 7.2 Hz, 2H).

[0341] Example 10: 4-(3-(4-methyl-7-(1 H-pyrazol-3-yl)-1 H-imidazo[4,5-d]thieno[3,2- b]pyridin-2-yl)propyl)morpholine (12)

[0342]

[0343] First Step: 2-(3-(Methyloxy)propyl)-l-(4-methoxybenzyl)-lH-imidazo[4,5- d]thieno[3,2-b]pyridine-5-oxide (10b)

[0344] Compound 10a (657 mg, 1.48 mmol), mCPBA (385 mg, 2.23 mmol) were added into 10 mL dichloromethane, the reaction was stirred at room temperature for 4 h. The reaction was poured into ice water, the organic phase was concentrated after dichloromethane extraction, compound 20b (534 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 460.2 [M+H] + .

[0345] Second Step: 2-(3-(Methyloxy)propyl)-l-(4-methoxybenzyl)-4-methyl-lH- imidazo[4,5-d]thieno[3,2-b]pyridine (20c)

[0346] Compound 10b (534 mg, 1.16 mmol) was added into 8 mL anhydrous tetrahydrofuran, then methyl magnesium bromide (691 mg, 5.81 mmol) was added, the reaction was stirred at 25 °C for 8 h. The reaction was poured into ice water, the organic phase was concentrated after dichloromethane extraction, compound 10c (388 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 458.2 [M+H] + .

[0347] Third Step: 3-(7-Bromo-l-(4-methoxybenzyl)-4-methyl-lH-imidazo[4,5-d]thieno[3,2- b]pyridin-2-yl)propan-l-ol (10d)

[0348] Compound 10c (388 mg, 0.85 mmol), N-bromosuccinimide (300 mg, 1.70 mmol) were added into 6 mL dichloromethane / glacial acetic acid (2: 1), the reaction was stirred at 25 °C for 2 h. The reaction was poured into ice water, dichloromethane was extracted, anhydrous sodium sulfate was dried overnight, the organic phase was concentrated after filtration, compound 10d (320 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 448.0 [M+H] + .

[0349] Fourth Step: 7-Bromo-2-(3-chloropropyl)-l-(4-methoxybenzyl)-4-methyl-lH- imidazo[4,5-d]thieno[3,2-b]pyridine (10e)

[0350] Compound 10d (237 mg, 0.53 mmol), DMF (4 mg, 0.05 mmol) were added into 4 mL of a mixed solvent of dichlorosulfoxide / tetrahydrofuran (1 : 1), and stirred at room temperature for 12 h. After the solvent was evaporated, compound 10e (195 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 466.0 [M+H] + .

[0351] Step 5: 4-(3-(7-bromo-1-(4-methoxybenzyl)-4-methyl-1H-imidazo[4,5-d]thieno[3,2- b]pyridin-2-yl)propyl)morpholine (10f)

[0352] Compound 10e (195 mg, 0.42 mmol), morpholine (73 mg, 0.84 mmol), tetrabutylammonium iodide (29 mg, 0.08 mmol), DIPEA (109 mg, 0.84 mmol) were added into 5 mL of DMF, and heated to 100 °C for 12 h under N2protection. After the solvent was evaporated, compound 10f (170 mg) was obtained by flash column chromatography (eluent system A).

[0353] MS (ESI, m / z): 517.1 [M+H] + .

[0354] Step 6: 4-(3-(1-(4-methoxybenzyl)-4-methyl-7-(1H-pyrazol-3-yl)-1H-imidazo[4,5- d]thieno[3,2-b]pyridin-2-yl)propyl)morpholine (10g)

[0355] Compound 10f (170 mg, 0.33 mmol), (1H-pyrazol-3-yl)boronic acid ester (128 mg, 0.66 mmol), Pd(dppf)Cl2(57 mg, 0.07 mmol), sodium carbonate (56 mg, 0.66 mmol) were added into 5 mL of a mixed solvent of DMF and 1 mL of water, and stirred at 110 °C for 6 h under N2protection. After filtration with diatomite, the filtrate was concentrated under reduced pressure, and compound 10g (55 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 503.2 [M+H] + .

[0356] Step 7: 4-(3-(4-methyl-7-(1H-pyrazol-3-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridin-2- yl)propyl)morpholine (12)

[0357] Compound 10g (55 mg, 0.11 mmol) was taken in 5 mL of trifluoroacetic acid, stirred at 65 °C for 10 hr, concentrated under reduced pressure, dissolved again in methanol, pH was adjusted to 8-9 using saturated sodium bicarbonate, filtered, concentrated the filtrate and purified by Prep-HPLC (Elution condition 1) to get compound 12 (13 mg).

[0358] MS (ESI, m / z): 383.2 [M+H] + .

[0359] 1 H NMR (DMSO-d6, 400 MHz) δ 13.03 (s, 2H), 7.78-7.83 (m, 2H), 6.81 (s, 1H), 3.53-3.55 (m, 4H), 2.90-2.92 (m, 2H), 2.73 (s, 3H), 2.34-2.35 (m, 6H), 1.95-2.02 (m, 2H).

[0360] Example 11: 4-cyclopropyl-l-methyl-2-(3-(4-methylpiperazin-l-yl)propyl)-7-(lH- pyrazol-3-yl)-lH-imidazo[4,5-d]thieno[3,2-b]pyridine (13)

[0361]

[0362] First Step: 2-(3-(benzyloxy)propyl)-4-cyclopropyl-l-methyl-lH-imidazo[4,5- d]thieno[3,2-b]pyridine (11a)

[0363] Compound Id (200 mg, 0.56 mmol), isopropyl magnesium bromide (410 mg, 2.83 mmol) were taken in 5 mL of dry tetrahydrofuran and stirred at 25 °C for 16 h. The reaction was poured into ice cold water, the organic layer was concentrated and purified by flash column chromatography (eluent system A) to get compound 11a (150 mg). MS (ESI, m / z): 378.2 [M+H] + .

[0364] Second Step: 2-(3-(benzyloxy)propyl)-7-bromo-4-cyclopropyl-l-methyl-lH-imidazo[4,5- d]thieno[3,2-b]pyridine (11b)

[0365] Compound 11a (150 mg, 0.40 mmol), N-bromosuccinimide (106 mg, 0.60 mmol) were taken in 10 mL of chloroform / glacial acetic acid (1:1) and stirred at room temperature for 16 h.

[0366] The reaction solution was poured into ice water, extracted with dichloromethane, and the organic phase was concentrated and purified by flash column chromatography (eluent system A) to obtain compound 11b (140 mg). MS (ESI, m / z): 456.1 [M+H] + .

[0367] Third step: 3-(7-bromo-4-cyclopropyl-l-methyl-lH-imidazo[4,5-d]thieno[3,2- b]pyridin-2-yl)propyl-l-ol (11c)

[0368] Compound 11b (100 mg, 0.22 mmol) was added to 5 mL of trifluoroacetic acid, heated to 80 °C and stirred for 16 h. The reaction solution was concentrated, redissolved in 3 mL of MeOH, and the pH was adjusted to 9.0 with 2N sodium hydroxide solution, and stirred for 30 min. Purification by flash column chromatography (eluent system A) gave compound 11c (70 mg). MS (ESI, m / z): 366.0 [M+H] + .

[0369] Fourth step: 7-bromo-2-(3-chloropropan-l-yl)-4-cyclopropyl-l-methyl-lH- imidazo[4,5-d]thieno[3,2-b]pyridine (11d)

[0370] Compound 11c (70 mg, 0.19 mmol) and 5 mL of dichlorosulfoxide were added to 5 mL of dichloromethane, and stirred at room temperature for 3 h. After the solvent was evaporated, purification by flash column chromatography (eluent system A) gave compound 11d (55 mg). MS (ESI, m / z): 386.0 [M+H] + .

[0371] Fifth step: 7-bromo-4-cyclopropyl-l-methyl-2-(3-(4-methylpiperazin-l-yl)propyl)- lH-imidazo[4,5-d]thieno[3,2-b]pyridine (11e)

[0372] Compound 11d (50 mg, 0.13 mmol), N-methylpiperazine (65 mg, 0.65 mmol), 18-crown-6 (14 mg, 0.06 mmol), and TEA (66 mg, 0.65 mmol) were added to 5 mL of toluene, and heated to 100 °C under N2protection for 16 h. After the solvent was evaporated, purification by flash column chromatography (eluent system A) gave compound 11e (30 mg). MS (ESI, m / z): 448.1 [M+H] + .

[0373] Step 6: 4-cyclopropyl-1-methyl-2-(3-(4-methylpiperazin-1-yl)propyl)-7-(1H- pyrazol-3-yl)-1H-imidazo[4,5-d]thieno[3,2-b]pyridine (13)

[0374] Compound 11e (30 mg, 0.067 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 1H-pyrazole (26 mg, 0.13 mmol), Pd(dppf)Cl2(11 mg, 0.013 mmol), sodium carbonate (21 mg, 0.20 mmol) were added in 4 mL of DMF and 1 mL of water mixed solvent, heated to 110 °C under N2protection and stirred for 3 h. The solvent was concentrated under reduced pressure, and compound 13 (12 mg) was separated by Prep-HPLC separation and purification (elution condition 1).

[0375] MS (ESI, m / z): 436.2 [M+H] + .

[0376] 1 H NMR (DMSO-d6, 400 MHz) δ 13.04 (s, 1H), 7.84 (s, 2H), 6.86 (s, 1H), 3.98 (s, 3H), 2.98-2.90 (m, 2H), 2.86-2.77 (m, 1H), 2.49-2.11 (m, 10H), 2.08 (s, 3H), 2.02-1.93 (m, 2H), 1.22-1.11 (m, 2H), 1.09-0.99 (m, 2H).

[0377] Example 12: 1-methyl-2-(3-(4-methylpiperazin-1-yl)propyl)-7-(1H-pyrazol-3-yl)-4- (trifluoromethyl)-1H-imidazo[4,5-c]quinoline (22)

[0378]

[0379] First step: 2-(3-(benzyloxy)propyl)-7-bromo-1-methyl-4-(trifluoromethyl)-1H- imidazo[4,5-c]quinoline 5-oxide (12a)

[0380] Compound 6b (1.9 g, 4.46 mmol), trifluoromethyltrimethylsilane (1.9 g, 13.37 mmol), cesium fluoride (177 mg, 2.67 mmol), 50 mL of anhydrous tetrahydrofuran were added into a reaction flask, and the mixture was stirred at 0 °C for 12 h. The reaction solution was poured into ice water, extracted with dichloromethane, and the organic phase was concentrated. Compound 12a (1.1 g) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 494.1 [M+H] + .

[0381] Second step: 2-(3-(benzyloxy)propyl)-7-bromo-1-methyl-4-(trifluoromethyl)-1H- imidazo[4,5-c]quinoline (12b)

[0382] Compound 12a (500 mg, 1.01 mmol), zinc powder (106 mg, 0.60 mmol), ammonium chloride (106 mg, 0.60 mmol), 5 mL of tetrahydrofuran, 5 mL of water were added into a reaction flask, and the mixture was stirred at room temperature for 3 h. The reaction solution was poured into ice water, extracted with dichloromethane, and the organic phase was concentrated. Compound 12b (300 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 478.1 [M+H] + .

[0383] Third step: 3-(7-bromo-1-methyl-4-(trifluoromethyl)-1H-imidazo[4,5-c]quinolin-2-yl)propyl-1-ol (12c)

[0384] Compound 12b (300 mg, 0.63 mmol) was added into 5 mL of trifluoroacetic acid, and the mixture was stirred at 80 °C for 16 h. The reaction solution was concentrated, redissolved in 3 mL of MeOH, and the pH was adjusted to 9.0 with 2N sodium hydroxide solution. The mixture was stirred for 30 min. Compound 12c (215 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 388.0 [M+H] + .

[0385] Fourth step: 7-bromo-2-(3-chloropropan-1-yl)-1-methyl-4-(trifluoromethyl)-1H-imidazo[4,5-c]quinoline (12d)

[0386] Compound 12c (215 mg, 0.55 mmol), N,N-dimethylformamide (20 mg, 0.277 mmol), 3 mL of dichlorosulfoxide, 3 mL of dichloromethane were added into a reaction flask, and the mixture was stirred at room temperature for 3 h. The solvent was evaporated, and compound 12d (180 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 408.0 [M+H] + .

[0387] Step 5: 7-Bromo-1-methyl-2-(3-(4-methylpiperazin-1-yl)propyl)-4- (trifluoromethyl)-1H-imidazo[4,5-c]quinoline (12e)

[0388] Compound 12d (70 mg, 0.17 mmol), N-methylpiperazine (100 mg, 0.86 mmol), tetrabutylammonium iodide (13 mg, 0.04 mmol), TEA (87 mg, 0.86 mmol) were added into 5 mL of toluene, heated to 100 °C for 16 h under N2protection. The solvent was rotary evaporated, and compound 12e (60 mg) was obtained by flash column chromatography (eluent system A). MS (ESI, m / z): 470.1 [M+H] + .

[0389] Step 6: 1-Methyl-2-(3-(4-methylpiperazin-1-yl)propyl)-7-(1H-pyrazol-3-yl)-4- (trifluoromethyl)-1H-imidazo[4,5-c]quinoline (22)

[0390] Compound 12e (60 mg, 0.13 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 1H-pyrazole (50 mg, 0.26 mmol), Pd(dppf)Cl2(31 mg, 0.038 mmol), sodium carbonate (41 mg, 0.38 mmol) were added into 4 mL of DMF and 1 mL of water mixed solvent, heated to 110 °C for 3 h under N2protection. After the reaction was completed, the reaction solvent was concentrated to dryness under reduced pressure, and compound 22 (18 mg) was separated and purified by Prep-HPLC (elution condition 1).

[0391] MS (ESI, m / z): 458.2 [M+H] + .

[0392] 1 H NMR (DMSO-d6, 400 MHz) δ 13.58 (s, 0.25H), 13.07 (s, 0.75H), 8.77-8.58 (m, 2H), 8.40-8.31 (m, 0.75H), 8.28-8.18 (m, 0.25H), 7.88 (s, 0.75H), 7.62 (s, 0.25H), 7.09-7.01 (m, 1H), 4.27 (s, 3H), 3.11-3.01 (m, 2H), 2.45-1.88 (m, 15H).

[0393] Biological evaluation

[0394] Experimental Example 1. Agonistic effect of the compounds of the present application on the expression of IL-1β in THP-1 cells after PMA-induced differentiation.

[0395] In this experiment, the HTRF (homogeneous time-resolved fluorescence) detection method was used to test the effect of the compounds of the present application on the level of the NLRP3 downstream cytokine IL-1β, so as to evaluate the agonistic effect of the compounds on the hNLRP3 inflammasome or the hNLRP3 inflammasome pathway at the cellular level.

[0396] Reagents: RPMI 1640 (Hyclone); heat-inactivated FBS (fetal bovine serum) (Gibco); PMA (tetradecanoyl phorbol acetate) (Bi Yun Tian)

[0397] Cells: THP-1 (Nanjing Kebai)

[0398] Kit: IL-1β assay kit (CISBIO)

[0399] Experimental steps:

[0400] 1) THP-1 cells in the logarithmic growth phase were inoculated in a T75 culture flask at a density of 5×10 5 cells / well, and were cultured in a cell incubator at 37°C and 5% CO2 for 24 h, and then were induced to become adherent macrophages by 1 μM PMA. The culture medium was RPMI 1640 containing 10% heat-inactivated FBS and 0.05 mM β-mercaptoethanol.

[0401] 2) After the cells were induced to culture for 24 h, the adherent cells were trypsinized, centrifuged at 1000 rpm for 5 min, and the supernatant was removed, and the cell density was resuspended to 2×10 6 cells / mL using RPMI 1640 medium containing 2% heat-inactivated FBS, and 50 μL / well of the cell resuspension was plated in a 96-well plate, and the number of cells per well was 1×10 5 .

[0402] 3) An appropriate amount of 10 mM DMSO solution of the test compound was prepared into 2× test concentration using RPMI 1640 medium containing 2% heat-inactivated FBS, and 50 μL / well of the dilution was added to the cells in the 96-well plate, and after being mixed well, the 96-well plate was placed in a cell incubator at 37°C and 5% CO2 for 6 h, and the supernatant was collected, and the level of IL-1β was measured according to the IL-1β detection kit instructions.

[0403] 4) EC 50 was fitted by the GraphPad software log (agonist) vs. response--Variable slope four-parameter method. The results are shown in Table 1.

[0404] Experimental Example 2. Agonistic effect of the compounds of the present application on the expression of IL-1β in PMA-induced differentiated THP1 cells (THP1- def NLRP3 cells) lacking NLRP3

[0405] This experiment used the HTRF (homogeneous time-resolved fluorescence) detection method to test the effect of the compounds of the present application on the IL-1β level in THP1- def NLRP3 cells, in order to assess the specificity of the compounds for the agonism of the hNLRP3 inflammasome or the hNLRP3 inflammasome pathway.

[0406] Reagents used: as described in Experimental Example 1

[0407] Cells: THP1- def NLRP3 (InvivoGen)

[0408] Kit: IL-1β assay kit (CISBIO)

[0409] Experimental procedure:

[0410] 1) THP1- def NLRP3 cells in logarithmic growth phase were seeded in T75 flasks at a density of 5 x 10 5 cells / well, and incubated at 37°C in a 5% CO2 incubator for 24 h, after which the THP1- def NLRP3 suspension cells were induced with 1 μM PMA to become adherent macrophages. The culture medium was RPMI 1640 containing 10% heat-inactivated FBS and 0.05 mM β-mercaptoethanol.

[0411] 2) After the cells were induced to culture for 24 h, the adherent cells were trypsinized, centrifuged at 1000 rpm for 5 min, and then the supernatant was removed, and the cells were resuspended to a density of 2 x 10 6 cells / mL using RPMI 1640 medium containing 2% heat-inactivated FBS, and 50 μL / well of the cell resuspension was plated in a 96-well plate, with a cell number of 1 x 10 5 cells / well.

[0412] 3) An appropriate amount of a 10 mM DMSO solution of the test compound was prepared to a 2x test concentration using RPMI 1640 medium containing 2% heat-inactivated FBS, and 50 μL / well of the dilution was added to the cells in the 96-well plate, which was mixed thoroughly, and then the plate was incubated at 37°C in a 5% CO2 cell incubator for 6 h, after which the supernatant was collected, and the IL-1β level was measured according to the IL-1β detection kit instructions.

[0413] 4) EC50 The slope was fitted using the four-parameter method of log(agonist) vs. response-variable slope in GraphPad software. The results are shown in Table 1.

[0414] Experimental Example 3: The agonistic effect of the compound of this invention on hTLR7

[0415] This experiment tested the activation effect of the compound of the present invention on the TLR7 signaling pathway by detecting luciferase in HEK-hTLR7-NF-κB-reporter cells, in order to evaluate the specificity of the compound on the NLRP3 pathway agonist effect.

[0416] Reagents: DMEM (High glucose); FBS (Fetal Bovine Serum) (Gibco); Bright-Glo TM Luciferase Detection Kit (Promega)

[0417] Cells: HEK-hTLR7-NF-κB-Luciferase gene cells (human TLR7NF-κB-luciferase reporter gene cells) (Nanjing Kebai)

[0418] 1) Trypsin-digest HEK-hTLR7-NF-κB-Luciferase cells in the logarithmic growth phase and resuspend them in culture medium to a concentration of 2×10⁻⁶. 6 Add 50 μL of cell resuspension per well to a 96-well plate at a concentration of 1 × 10⁶ cells / mL, resulting in 1 × 10⁶ cells per well. 6 Take an appropriate amount of 10 mM DMSO solution of the test compound, prepare a 2× test concentration with culture medium, add 50 μL / well to the cells of a 96-well plate, and incubate the 96-well plate in a 37°C, 5% CO2 incubator for 16 h. The culture medium is DMEM (High glucose) containing 10% FBS.

[0419] 2) After cell incubation, add 100 μL / well Bright-Glo TM Luciferase assay reagent, incubate at room temperature for 5 min, and read the relative luciferase unit (RLU) using a microplate reader.

[0420] 3) The stimulatory effect of the tested compounds on hTLR7 (EC) 50 The slope was fitted using the four-parameter method of log(agonist) vs. response-variable slope in GraphPad software. The results are shown in Table 1.

[0421] Experimental Example 4: Agonistic effect of the compounds of the application on hTLR8

[0422] This experiment tests the activation of the TLR8 signaling pathway by the compounds of the application by measuring the amount of secreted alkaline phosphatase in the HEK-Blue cell line, in order to assess the specificity of the compounds for the agonistic effect on the NLRP3 pathway.

[0423] Reagents: DMEM (High glucose); FBS (Fetal Bovine Serum) (Gibco); QUANTI-Blue / InvivoGen / rep-qb2;

[0424] Cells: HEK-Blue TM hTLR8 cells (human TLR 8 cells) (InvivoGen)

[0425] Experimental procedure:

[0426] 1) The HEK-Blue hTLR8 cells in the logarithmic growth phase were trypsinized, resuspended in medium to a concentration of 2 x 10 TM hTLR8 cells were trypsinized, resuspended in medium to a concentration of 2 x 10 6 μL / well of cell suspension in a 96-well plate; an appropriate amount of 10 mM DMSO solution of the test compound was prepared to 2 x the test concentration with medium, and 50 μL / well was added to the cells in the 96-well plate, and the 96-well plate was placed in a 37°C, 5% CO2 incubator for 16 h. The culture medium was DMEM (High glucose) containing 10% FBS.

[0427] 2) After the incubation of the cells was completed, 10 μL of the cell culture supernatant was transferred to a 96-well plate, 90 μL / well of QUANTI-Blue detection solution was added, and the plate was incubated at 37°C for 3 h, and the OD 620 reading was taken on a microplate reader. The results are shown in Table 1.

[0428] Table 1. Agonistic activity and specificity of the compounds of the application on hNLRP3

[0429]

[0430] The results show that the compounds of the application (e.g. compounds 1, 2, 3, 5, 6, 7, 8, 9, 10, 11, 13, 22) have a significant agonistic effect on the expression of IL-1β in PMA-induced differentiated THP-1 cells, while having no effect on the expression of IL-1β in THP1- defThe IL-1β expression in NLRP3 cells was not excited at the highest compound test concentration (27 μM), and there was no significant activation of hTLR7 and hTLR8 at the highest compound test concentration (100 μM). In summary, the compounds (e.g., compounds 1, 2, 3, 5, 6, 7, 8, 9, 10, 11, 13, 22) of the present application have obvious agonistic activity on hNLRP3 and its signaling pathway.

[0431] Experimental Example 5: hERG test

[0432] Using Predictor TM The hERG Fluorescence Polarization Assay Kit (manufacturer: ThermoFisher) was used to test the inhibition of hERG potassium ion channel by the compounds (concentration: 10 μM) according to the kit instructions, and the test results are shown in Table 2.

[0433] Table 2. Inhibition test results of compounds on hERG

[0434] Compound No. IC 50 (μM) 1 >10 2 >10 6 >10 7 >10 11 >10

[0435] The results show that the compounds represented by compounds 1, 2, 6, 7 and 11 have no significant inhibition on hERG, and have little possibility of causing prolongation of cardiac QT interval.

[0436] Experimental Example 6: CYP enzyme inhibition test

[0437] CYP450 is the most important enzyme system in drug metabolism, and the enzymes involved in metabolism interact with drugs, of which the most important are CYP1A2, CYP2D6 and CYP3A4.

[0438] In the test for inhibition of CYP450 enzymes by fluorescence method, P450-Glo TM CYP1A2 Screening System, CYP2D6 Cyan Screening Kit and CYP3A4 Red Screening Kit were used to determine the inhibition activity of the compounds on CYP1A2, CYP2D6 and CYP3A4, respectively, according to the kit instructions. The test results are shown in Table 3.

[0439] Table 3. Inhibition test results of compounds on CYP enzymes

[0440]

[0441] Results show that the compounds represented by compounds 2, 3, 6, 9, 10, 11 and 12 have no obvious inhibitory effect on CYP1A2, CYP2D6 and CYP3A4 enzymes.

[0442] In addition to those described herein, various modifications and alterations of the application will become apparent to those skilled in the art from the foregoing description. It is intended that all such modifications and alterations be considered as falling within the scope of the appended claims. Each of the references cited herein (including all patents, patent applications, journal articles, books, and any other publications) is incorporated herein in its entirety by reference.

Claims

1. A compound having a structure according to Formula X: ###0001### or a pharmaceutically acceptable salt thereof. wherein: X 2 is N; R 6 is not present; R 2 selected from C 1-4 alkyl; -L-R 3 selected from the group consisting of: R 4 selected from C 1-4 alkyl and C 3-6 cycloalkyl; selected from wherein, R 1 selected from C 3-6 cycloalkyl, 4-7 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl; said C 3-6 cycloalkyl, 4-7 membered heterocyclyl, C 6-12 aryl, 5-10 membered heteroaryl can be optionally substituted with one or more of the following substituents: halo, CN, C 1-4 alkyl, C 3-6 cycloalkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 hydroxyalkyl; X 1 is CH.

2. The compound or pharmaceutically acceptable salt thereof of claim 1, wherein, R 1 selected from C 6-12 aryl and 5-10 membered heteroaryl, said C 6-12 aryl, 5-10 membered heteroaryl can be optionally substituted with one or more of the following substituents: halo, CN, C 1-4 alkyl, C 3-6 cycloalkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 hydroxyalkyl.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, R 2 is selected from the group consisting of methyl, ethyl, propyl and butyl.

4. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 4 is selected from the group consisting of methyl, isopropyl and cyclopropyl.

5. The compound according to any one of claims 1-4, or a pharmaceutically acceptable salt thereof, having a structure according to Formula VI: ###0004### R 1 , R 2 , R 3 , R 4 , X 1 , L is as defined in any one of claims 1 to 4.

6. A pharmaceutical composition comprising a compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, optionally further comprising one or more pharmaceutically acceptable carriers.

7. A pharmaceutical preparation comprising a compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 6.

8. Use of a compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 6, for the manufacture of a medicament for the prevention, alleviation and / or treatment of a disease associated with NLRP3 inflammasome activity.

9. Use of a compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 6, for the manufacture of a medicament for the prevention, alleviation and / or treatment of a disease of abnormal cell proliferation.

10. Use of a compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 6, for the manufacture of a medicament for the prevention, alleviation and / or treatment of a neoplastic disease.

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