A novel pyridopyrazole heterocyclic compound and its application

Abstract

The present invention provides a novel pyridopyrazole heterocyclic compound, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, and a related pharmaceutical composition thereof. The present invention also provides the use of such a compound and a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, and a related pharmaceutical composition thereof in the preparation of a drug for treating and preventing medical conditions and diseases by inhibiting NLRP3.

Description

Technical Field

[0001] The present invention relates to pyridopyrazole heterocyclic compounds, and specifically to compounds represented by formula (I), pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, and related pharmaceutical compositions thereof. The present invention also relates to the use of such compounds and pharmaceutically acceptable salts thereof, solvates thereof, prodrugs thereof, isomers thereof, and related pharmaceutical compositions thereof in the preparation of drugs for treating and preventing related diseases, especially in the treatment and prevention of medical conditions and diseases by inhibiting NLRP3. Background Art

[0002] NOD-like receptors (NLRs) protein inflammasome is a type of pattern recognition receptors (PRRs), which together with Toll like receptors (TLRs), retinoic acid-inducible gene I (RIG-I) like receptors (RLRs) and C-type lectin receptors (CLRs) constitute the pattern recognition receptors of innate immunity [He Shaoping et al., Journal of Animal Nutrition, 2017, 29(11): 3844-3851].

[0003] NLRP3 (NACHT, LRR and PYD domains-containing protein 3) inflammasome is an important member of NLRs protein inflammasome. It is a macromolecular multiprotein complex with a relative molecular mass of about 700kD, which is composed of the intracellular innate immune receptor NLRP3, the adaptor protein ASC (apoptosis-associated speck-like protein containing a CARD) and the effector protein caspase-1 (cysteine ​​aspartate protease 1). The complex assembly can induce the maturation and secretion of proinflammatory factors such as IL-1β and IL-18, thereby promoting the occurrence of inflammatory response [Coll RC et al., Nat Med. 2015Mar; 21(3): 248-55].

[0004] NLRP3 is mainly expressed in immune cells such as dendritic cells, monocytes, macrophages, and neutrophils. Under normal circumstances, NLRP3 is combined with molecular chaperones such as heat shock protein 90 and is in a self-inhibited state. When stimulated by agonists from its own source (such as ATP, cholesterol crystals, urate crystals, calcium dihydrogen pyrophosphate crystals, etc.), agonists from the environment (such as aluminum particles, asbestos, silicon, alloy particles, ultraviolet damage, etc.) or pathogenic agonists (such as bacterial pore-forming toxins, flagellin, rod proteins, cell wall diacyl peptides, etc.), NLRP3 recruits the adaptor protein ASC through its N-terminal PYD to form the NLRP3 inflammasome with pro caspase-1. After the NLRP3 inflammasome is assembled, self-cleavage occurs to form caspase-1 with cleavage activity, which cleaves inactive pro IL-1β and pro IL-18 into active IL-1β and IL-18 and secretes them, allowing them to play a biological role, mediate immune response and inflammatory response, and maintain the body's immune homeostasis [Kelley N et al., Int. J. Mol. Sci. 2019, 20, 3328]. IL-1β signaling can induce the secretion of proinflammatory cytokines IL-6 and TNF, and IL-18 and IL-12 can synergistically induce Th1 cells, NK cells, and NKT cells to produce IFN-γ.

[0005] However, overactivation of NLRP3 and gene mutations can lead to severe inflammatory diseases. For example, persistent activation of NLRP3 caused by gene mutations is associated with familial cold-induced autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), and neonatal onset multi-system inflammatory disorder or chronic infantile neurologic cutaneous and articular syndrome (NOMID / CINCA) [Wilson SP et al., Postgrad Med. 2010 September; 122(5): 125–133].

[0006] NLRP3 is also involved in the occurrence and development of various types of diseases, such as multiple sclerosis (MS) [Olcum M et al., Adv Protein Chem Struct Biol. 2020; 119: 247-308], systemic lupus erythematosus (SLE) [Kahlenberg JM et al., Curr Opin Rheumatol. 2014 Sep; 26 (5): 475-81], rheumatoid arthritis (RA) [Guo C et al., Clin Exp Immunol. 2018 Nov; 194 (2): 231-243], inflammatory bowel disease (IBD) [Zhen Y et al., Front Immunol. 2019 Feb 28; 10: 276], blindness caused by visual pigment cell degeneration [Ildefonso et al., Front Immunol. 2019 Feb 28; 10: 276], and leukemia [Leukemia and Blindness]. CJ et al., Adv Exp Med Biol. 2016; 854: 59-65], acute lung injury [Grailer JJ et al., J Immunol. 2014 Jun 15; 192(12): 5974-83], contact hypersensitivity [ AI et al., Pharmaceutics. 2020 Sep 11; 12(9): 867], sepsis [Danielski, LG et al., Inflammation 43, 24–31 (2020)], aging [Youm, YH et al., Cell Metab. 18, 519–532 (2013)], Parkinson's disease [Heneka MT et al., Nat Rev Immunol. 2014 Jul; 14(7): 463-77], non-alcoholic fatty liver disease [Xingyong Wan et al., Can J Gastroenterol Hepatol 2016; 2016: 6489012.], etc. It has been reported that the NLRP3 inflammasome is involved in the treatment of EAE mice with β-interferon through the experimental autoimmune encephalomyelitis (EAE) model, and in vivo experiments in mice have demonstrated the correlation between the NLRP3 inflammasome and Alzheimer's disease [Heneka, M. et al., Nature 493, 674–678 (2013)].

[0007] NLRP3 inflammasome is also involved in some common metabolic diseases, such as type 2 diabetes, gout, obesity, atherosclerosis, etc. [Wen, H. et al., Nat Immunol 13, 352–357 (2012)]. In addition, NLRP3 inflammasome is also related to the occurrence and development of tumors. For example, the myeloid suppressor cells induced by the chemotherapy drugs gemcitabine and 5-fluorouracil cause increased lysosomal permeability through cathepsin B, resulting in the release of cathepsin B and interaction with NLRP3, thereby activating the NLRP3 inflammasome, and ultimately weakening the anti-tumor effect of chemotherapy drugs through IL-17 [Bruchard, M. et al., Nat Med 19, 57–64 (2013)]. Therefore, NLRP3 inflammasome is a potential target for the treatment of these diseases.

[0008] The current clinical treatment of NLRP3 inflammasome-related diseases is mainly biological agents targeting IL-1, including the IL-1β neutralizing antibody Canakinumab, the recombinant protein IL-1R antagonist Anakinra, and the soluble inducible receptor protein Rilonacept that can bind to IL-1α and IL-1β. These drugs have been confirmed to play a role in the treatment of CAPS and have been used in clinical trials for other IL-1β-related diseases [Dinarello CA et al., Semin Immunol. 2013 Dec 15; 25(6): 469-84].

[0009] However, existing IL-1-related biological agents have their own limitations. For example, they cannot pass through the blood-brain barrier and therefore cannot effectively reduce central nervous system inflammation, which leaves room for exploration of small molecule drugs. In addition, IL-1 is not the only effector molecule produced after activation of the NLRP3 inflammasome (there are also IL-18, etc.), and drugs targeting IL-1 cannot effectively treat all NLRP3 inflammasome-related diseases. In addition, IL-1 not only comes from the NLRP3 inflammasome, but the activation of other inflammasomes such as NLRP1 and NLRC4 can also stimulate the secretion of IL-1. Treatment targeting IL-1 or its receptors will increase the risk of infection because some cytokines are necessary to resist pathogenic viruses and bacteria.

[0010] In contrast, drug therapy targeting NLRP3 inflammasome is a better choice, which can fundamentally inhibit diseases mediated by IL-1, IL-18 and HMGB1, achieve more specific therapeutic effects, and reduce the risk of infection. Therefore, it is more valuable to target NLRP3 inflammasome to treat related inflammatory diseases.

[0011] Currently, there are no drugs on the market that specifically inhibit NLRP3 inflammasomes. It has been reported that several small molecule compounds can effectively inhibit the activation of NLRP3 inflammasomes, including glibenclamide [Lamkanfi M et al., J Cell Biol. 2009 Oct 5; 187(1): 61-70], parthenolide [Juliana C et al., J. Biol Chem. 285(13): 9792-9802], 3,4-methylenedioxy-β-nitrostyrene [He Y et al., J Biol Chem. 2014 Jan 10; 289(2): 1142-50], dimethyl sulfoxide [Ahn H et al., Immunobiology. 2014 Apr; 219(4): 315-22], MCC950 [Coll RC et al., Nat Med. 2015 Mar; 21(3): 248-55], CY-09 [Jiang H et al., J Exp Med. 2017 Nov 6; 214(11): 3219-3238]、OLT1177

[25] 、Tranilast[Huang Y EMBO Mol Med. 2018 April; 10(4): e8689 et al.]、Oridonin[He, H. et al., Nat Commun 9, 2550(2018)]、Bay 11-7082[Juliana C et al., J. Biol Chem. 285(13): 9792–9802]、Fenamate[Daniels, M. et al., Nat Commun 7, 12504(2016)] etc. However, some of them have non-specific effects and weak activity. In addition, some compounds containing diarylsulfonylurea, sulfonylurea and sulfenyl imine have been disclosed as NLRP3 inhibitors (CN107428696A, CN110366549A, etc.).

[0012] Currently, there is still a need for new NLRP3 inflammasome activation-specific inhibitors with better activity, greater safety, better pharmacokinetic properties and better physicochemical properties. It is of great significance to find such molecules in order to effectively treat NLRP3 inflammasome-related diseases. Summary of the invention

[0013] The first aspect of the present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt, solvate, prodrug and isomer thereof:

[0014]

[0015] in:

[0016] A 1 , A 2, A 3 , A 4 Each independently selected from N or CR 3 ;

[0017] X is O, NH or N(CN);

[0018] Y is O or S;

[0019] R 1 Selected from: H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 heteroalkyl containing 1-2 heteroatoms, C3-C6 saturated heterocycloalkyl containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), C2-C6 alkenyl, C2-C6 alkynyl or -R 4 -R 5 , may be optionally substituted by one or more substituents selected from halogen atoms, amino groups, hydroxyl groups, C1-C3 alkyl groups, and C3-C6 cycloalkyl groups;

[0020] R 2 is selected from: phenyl or other C5-C6 heteroaryl substituted at the α-position and α'-position, and R 2 may optionally be further substituted;

[0021] R 3 Selected from: H, halogen, nitro, cyano, -OR 8 -R 9 , -R 10 -R 11 , -SR 13 、-(C=O)R 14 、-SO2NR 15 R 16 、-(P=O)R 17 R 18 、-NR 19 R 20 ;

[0022] R 4 Selected from: C1-C6 alkylene, C1-C6 heteroalkylene (the heteroatom may be oxygen, nitrogen or sulfur), each of which may be optionally substituted by one or more substituents selected from halogen atoms, amino groups, hydroxyl groups, C1-C3 alkyl groups, and C3-C6 cycloalkyl groups;

[0023] R 5Selected from: -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -O(C1-C6 alkyl), -NH(C3-C6 cycloalkyl), -N(C3-C6 cycloalkyl)2, -N(C1-C6 alkyl)(C3-C6 cycloalkyl), -O(C3-C6 cycloalkyl), C3-C6 cycloalkyl, C3-C6 saturated heterocyclic group containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), all of which may be optionally substituted by one or more substituents selected from halogen atoms, amino, hydroxyl, C1-C6 alkyl, C3-C6 cycloalkyl; R 5 It may also be selected from phenyl or C5-C6 heteroaryl, all of which may be optionally substituted by one or more halogen atoms and / or optionally substituted by one or more substituents independently selected from the following: cyano, C1-C6 alkyl, C3-C6 cycloalkyl, C3-C6 saturated heterocycloalkyl containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), C1-C6 haloalkyl, C3-C6 halocycloalkyl, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -O(C1-C6 alkyl), -S(C1-C6 alkyl) or -R 6 -R 7 ;

[0024] R 6 Selected from: C1-C6 alkylene, C1-C6 heteroalkylene (the heteroatom may be oxygen, nitrogen or sulfur);

[0025] R 7 Selected from: C3-C6 cycloalkyl, C3-C6 saturated heterocyclic group containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), all of which may be optionally substituted by one or more halogen atoms;

[0026] R 8 Selected from: a bond, a C1-C6 alkylene group, each of which may be optionally substituted by one or more halogen atoms;

[0027] R 9 Selected from: H, amino, hydroxyl, -NH(C1-C6 alkyl), -NH(C3-C6 cycloalkyl), -N(C1-C6 alkyl)2, -N(C3-C6 cycloalkyl)2, -N(C1-C6 alkyl)(C3-C6 cycloalkyl), -O(C1-C6 alkyl), -O(C3-C6 cycloalkyl), C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 heteroalkyl containing 1-2 heteroatoms, C3-C6 saturated heterocyclic group containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), all of which may be optionally substituted by one or more halogen atoms;

[0028] R 10is selected from: a bond, a C1-C6 alkylene group, a C1-C6 heteroalkylene group containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), each of which may be optionally substituted by one or more halogen atoms;

[0029] R 11 Selected from: cyano, amino, hydroxyl, -NH(C1-C6 alkyl), -NH(C3-C6 cycloalkyl), -N(C1-C6 alkyl)2, -N(C3-C6 cycloalkyl)2, -N(C1-C6 alkyl)(C3-C6 cycloalkyl), -O(C1-C6 alkyl), -O(C3-C6 cycloalkyl), C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 heteroalkyl containing 1-2 heteroatoms, C3-C6 saturated heterocyclic group containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), phenyl, C5-C6 heteroaryl, all of which may be optionally substituted by one or more halogen atoms;

[0030] R 12 , R 17 , R 18 Each is independently selected from: C1-C6 alkyl, C3-C6 cycloalkyl;

[0031] R 13 Selected from: H, C1-C6 alkyl, C1-C6 heteroalkyl containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), C3-C6 cycloalkyl, all of which may be optionally substituted with one or more halogen atoms, amino, hydroxyl, C1-C6 alkyl, C3-C6 cycloalkyl;

[0032] R 14 Selected from: C1-C6 alkyl, C3-C6 cycloalkyl, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -O(C1-C6 alkyl);

[0033] R 15 , R 16 Each is independently selected from: H, C1-C6 alkyl, C3-C6 cycloalkyl, R 15 , R 16 It can also form a saturated C3-C6 heterocyclic group with the connected nitrogen atom, and the heterocyclic group contains 1-2 heteroatoms. In addition to N, the other heteroatom can be sulfur, oxygen or nitrogen;

[0034] R 19 , R 20 Each independently selected from: H, -(C=O)R 22, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 heteroalkyl containing 1-2 heteroatoms, C3-C6 saturated heterocyclic group containing 1-2 heteroatoms, all of which may be optionally substituted by one or more substituents selected from halogen atoms, hydroxyl, amino, -NH(C1-C3 alkyl), -N(C1-C3 alkyl)2, C1-C6 alkyl, C3-C6 cycloalkyl, C3-C6 saturated heterocyclic group containing 1-2 heteroatoms, wherein the heteroatoms may be oxygen, nitrogen or sulfur; R 19 , R 20 It can also form a saturated C3-C6 heterocyclic group with the connected nitrogen atom, and the heterocyclic group contains 1-2 heteroatoms. In addition to N, the other heteroatom can be sulfur, oxygen or nitrogen;

[0035] R 21 , R 22 Each is independently selected from: C1-C6 alkyl, C3-C6 cycloalkyl, and each may be optionally substituted by one or more halogen atoms.

[0036] Preferably, wherein R 2 is phenyl or pyridyl optionally substituted at the α-position and α'-position, and R 2 may optionally be further substituted.

[0037] Preferably, Y is O.

[0038] Preferably, wherein R 2 Selected from:

[0039] in

[0040] B 1 , B 2 Each independently selected from: -CH2CH2CH2-, -CH2CH=CH-, -CH=CHCH2-, -CH2CH2O-, -OCH2CH2-, -CH2OCH2-, -CH2CH2CH2CH2- and -CH=CH-CH=CH-, and each may be optionally substituted by one or more halogen atoms and / or optionally substituted by one or two substituents independently selected from the following: oxo, -OH, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, C3-C6 halocycloalkyl, -O(C1-C6 alkyl), -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2;

[0041] R a , R g and R hEach is independently selected from: C1-C6 alkyl, C3-C6 cycloalkyl, -S(C1-C6 alkyl), phenyl, all of which may be optionally substituted by one or more halogen atoms or deuterium atoms and / or optionally substituted by one or two substituents independently selected from: oxo, cyano, -OH, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, C3-C6 halocycloalkyl, -O(C1-C6 alkyl), -O(C1-C6 haloalkyl);

[0042] R e , R f Each is independently selected from C1-C6 alkyl, C3-C6 cycloalkyl, -S(C1-C6 alkyl), phenyl, C5-C6 heteroaryl, and each may be optionally substituted by one or more halogen atoms or deuterium atoms and / or optionally substituted by one or two substituents independently selected from the following: oxo, cyano, -OH, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, C3-C6 halocycloalkyl, -O(C1-C6 alkyl), -O(C1-C6 haloalkyl);

[0043] R b , R d Each is independently selected from hydrogen, halogen, methoxy, cyano, nitro;

[0044] R c Selected from: H, halogen, cyano, nitro, C1-C6 alkyl, C3-C6 cycloalkyl, -(C=O)(C1-C6 alkyl), -CO2(C1-C6 alkyl), -CONH(C1-C6 alkyl), -(C=O)N(C1-C6 alkyl)2, -O(C1-C6 alkyl), all of which may be optionally substituted by one or more halogen atoms.

[0045] Preferably, B 1 and B 2 Each independently selected from:

[0046] -CH2CH2CH2-, -CH2CH2CH2CH2-, -CH2CH2O-, -OCH2CH2-, -CH2OCH2-;

[0047] R a , R g and R h Each is independently selected from isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio, phenyl, wherein the phenyl is optionally substituted by a halogen atom or a deuterium atom and / or is optionally substituted by one or two substituents independently selected from the following: cyano, -OH, -O(C1-C6 alkyl) and -O(C1-C6 haloalkyl);

[0048] Re , R f Each is independently selected from: isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio, phenyl and pyridyl, wherein pyridyl and phenyl may be optionally substituted by a halogen atom or a deuterium atom and / or optionally substituted by one or two substituents independently selected from: cyano, -OH, -O(C1-C6 alkyl) and -O(C1-C6 haloalkyl);

[0049] R b , R d Each is independently selected from: hydrogen, halogen, methoxy, cyano;

[0050] R c Selected from: H, halogen, cyano, C1-C6 alkyl, C3-C6 cycloalkyl, -(C=O)(C1-C6 alkyl), -CO2(C1-C6 alkyl), -CONH(C1-C6 alkyl), -(C=O)N(C1-C6 alkyl)2, -O(C1-C6 alkyl), all of which may be optionally substituted by one or more halogen atoms.

[0051] Preferably, X is O.

[0052] Preferably, wherein: A 1 , A 2 , A 3 and A 4 There are 0 or 1 N in .

[0053] More preferably, the compound has a structure shown in formula (II)-(VI):

[0054]

[0055] Where R 1 Selected from: H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 heteroalkyl containing 1-2 heteroatoms, C3-C6 saturated heterocycloalkyl containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur) or -R 4 -R 5 , may be optionally substituted by one or more substituents selected from halogen atoms, amino groups, hydroxyl groups, C1-C3 alkyl groups, and C3-C6 cycloalkyl groups;

[0056] R 2 Selected from:

[0057] Among them B 1 and B 2

[0058] Each independently selected from:

[0059] -CH2CH2CH2-, -CH2CH2CH2CH2-, -CH2CH2O-, -OCH2CH2-, -CH2OCH2-;

[0060] R a , R g and R h Each is independently selected from isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio, phenyl, wherein the phenyl is optionally substituted by a halogen atom or a deuterium atom and / or is optionally substituted by one or two substituents independently selected from the following: cyano, -OH, -O(C1-C6 alkyl) and -O(C1-C6 haloalkyl);

[0061] R e , R f Each is independently selected from: isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio, phenyl and pyridyl, wherein pyridyl and phenyl may be optionally substituted by a halogen atom or a deuterium atom and / or optionally substituted by one or two substituents independently selected from: cyano, -OH, -O(C1-C6 alkyl) and -O(C1-C6 haloalkyl);

[0062] R b , R d Each is independently selected from: hydrogen, halogen, methoxy, cyano;

[0063] R c Selected from: H, halogen, cyano, C1-C6 alkyl, C3-C6 cycloalkyl, -(C=O)(C1-C6 alkyl), -CO2(C1-C6 alkyl), -CONH(C1-C6 alkyl), -(C=O)N(C1-C6 alkyl)2, -O(C1-C6 alkyl), all of which may be optionally substituted by one or more halogen atoms;

[0064] R 3’ Selected from: H, halogen, cyano, -OR 8 R 9 , -R 10 -R 11 、-SR 13 、-NR 19 R 20 ;

[0065] R 4 Selected from: C1-C6 alkylene, C1-C6 heteroalkylene (the heteroatom may be oxygen, nitrogen or sulfur), each of which may be optionally substituted by one or more substituents selected from halogen atoms, amino groups, hydroxyl groups, C1-C3 alkyl groups, and C3-C6 cycloalkyl groups;

[0066] R5 Selected from: -NH(C1-C6 alkyl), -O(C1-C6 alkyl), -N(C1-C6 alkyl)2, C3-C6 cycloalkyl, -NH(C3-C6 cycloalkyl), -N(C3-C6 cycloalkyl)2, -N(C1-C6 alkyl)(C3-C6 cycloalkyl), -O(C3-C6 cycloalkyl), C3-C6 saturated heterocyclic group containing 1-2 heteroatoms (the heteroatoms are selected from oxygen, sulfur or nitrogen), all of which may be optionally substituted by one or more substituents selected from halogen atoms, amino, hydroxyl, C1-C6 alkyl, C3-C6 cycloalkyl; R 5 It may also be selected from phenyl or C5-C6 heteroaryl, all of which may be optionally substituted by one or more halogen atoms and / or optionally substituted by one or more substituents independently selected from the following: cyano, C1-C6 alkyl, C3-C6 cycloalkyl, C3-C6 saturated heterocycloalkyl containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, -O(C1-C6 alkyl), -S(C1-C6 alkyl) or -R 6 -R 7 ;

[0067] R 6 Selected from: C1-C6 alkylene;

[0068] R 7 Selected from: C3-C6 cycloalkyl, C3-C6 saturated heterocyclic group containing 1-2 heteroatoms (the heteroatoms are selected from oxygen, sulfur or nitrogen);

[0069] R 8 Selected from: a bond, a C1-C6 alkylene group;

[0070] R 9 Selected from: H, amino, hydroxyl, -NH(C1-C6 alkyl), -NH(C3-C6 cycloalkyl), -N(C1-C6 alkyl)2, -N(C3-C6 cycloalkyl)2, -N(C1-C6 alkyl)(C3-C6 cycloalkyl), -O(C1-C6 alkyl), -O(C3-C6 cycloalkyl), C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 heteroalkyl containing 1-2 heteroatoms, C3-C6 saturated heterocyclic group containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), all of which may be optionally substituted by one or more halogen atoms;

[0071] R 10 is selected from: a bond, a C1-C6 alkylene group, a C1-C6 heteroalkylene group containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur);

[0072] R 11Selected from: cyano, amino, hydroxyl, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 heteroalkyl containing 1-2 heteroatoms (the heteroatoms may be oxygen, nitrogen or sulfur), piperazinyl, morpholinyl, tetrahydropyrrolyl, tetrahydrofuranyl, Tetrahydropyranyl, piperidinyl, phenyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, -NH(C1-C6 alkyl), -NH(C3-C6 cycloalkyl), -N(C1-C6 alkyl)2, -O(C1-C6 alkyl), all of which may be optionally substituted with one or more halogen atoms;

[0073] R 13 Selected from: H, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 heteroalkyl containing 1-2 heteroatoms (the heteroatoms are oxygen, nitrogen or sulfur);

[0074] R 19 , R 20 Each is independently selected from: H, C1-C6 alkyl, C3-C6 cycloalkyl, -(C1-C6 alkylene)-NH-(C1-C3 alkyl), -(C1-C6 alkylene)-N(C1-C3 alkyl)2; R 19 , R 20 It can also form a saturated C3-C6 heterocyclic group with the nitrogen atom to which it is connected. The heterocyclic group contains 1-2 heteroatoms. In addition to N, the other heteroatom is sulfur, oxygen or nitrogen.

[0075] Preferably, wherein R 1 Selected from: -CH2F, -CHF2, -CF3, C1-C6 alkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxy-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, -R 4 -R 5 Or the following groups:

[0076]

[0077] R 2 Selected from:

[0078] Among them B 1 and B 2 Each independently selected from:

[0079] -CH2CH2CH2-, -CH2CH2CH2CH2-, -CH2CH2O-, -OCH2CH2-, -CH2OCH2-;

[0080] R a , R g and R hEach is independently selected from isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio, and phenyl;

[0081] R e , R f Each is independently selected from isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio, phenyl, pyridyl, 2-chloropyridin-4-yl, 2-cyanopyridin-4-yl, 2-methoxypyridin-4-yl;

[0082] R b , R d Each independently selected from: H, F, Cl, CN;

[0083] R c Selected from: H, F, Cl, CN;

[0084] R 3’ Selected from: H, F, Cl, CN, -OR 8 R 9 , -R 10 -R 11 or -NR 19 R 20 ;

[0085] R 4 Selected from: C1-C6 alkylene, amino-substituted C1-C6 alkylene, hydroxy-substituted C1-C6 alkylene;

[0086] R 5 Selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -NH(C1-C6 alkyl), -O(C1-C6 alkyl), -N(C1-C6 alkyl)2, -NH(C3-C6 cycloalkyl) or the following groups:

[0087]

[0088] R 8 , R 10 Each independently selected from: a bond, a C1-C6 alkylene group;

[0089] R 9 Selected from: H, amino, hydroxyl, C1-C6 alkyl, C3-C6 cycloalkyl, -NHCH3, -N(CH3)2, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, -CF3, CHF2;

[0090] R 11 Selected from: C1-C6 alkyl, C3-C6 cycloalkyl, amino, hydroxyl, -CHF2, -CF3, piperazinyl, morpholinyl, piperidinyl, tetrahydropyranyl, tetrahydropyrrolyl, tetrahydrofuranyl, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2;

[0091] R 19 , R 20 Each is independently selected from: H, C1-C6 alkyl, -(C1-C4 alkylene)-NH-(C1-C3 alkyl), -(C1-C4 alkylene)-N(C1-C3 alkyl)2; R 19 , R 20 It can also form a saturated C3-C6 heterocyclic group with the adjacent nitrogen atom, the heterocyclic group contains 1-2 heteroatoms, except for N, the other heteroatom is sulfur, oxygen or nitrogen;

[0092] R 23 It is H, C1-C6 alkyl.

[0093] More preferably, the compound has a structure shown in formula (III):

[0094]

[0095] Where R 1 Selected from: -CH2F, -CHF2, -CF3, C1-C6 alkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxy-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, -R 4 -R 5 Or the following groups:

[0096]

[0097] R 2 Selected from:

[0098] Among them B 1 and B 2 Each independently selected from:

[0099] -CH2CH2CH2-, -CH2CH2CH2CH2-, -CH2CH2O-, -OCH2CH2-, -CH2OCH2-;

[0100] R a , R g and R h Each is independently selected from isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio, and phenyl;

[0101] R e , R fEach is independently selected from isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio, phenyl, pyridyl, 2-chloropyridin-4-yl, 2-cyanopyridin-4-yl, 2-methoxypyridin-4-yl;

[0102] R b , R d Each independently selected from: H, F, Cl, CN;

[0103] R c Selected from: H, F, Cl, CN;

[0104] R 3’ Selected from: H, F, Cl, CN, -OR 8 R 9 , -R 10 -R 11 or -NR 19 R 20 ;

[0105] R 4 Selected from: C1-C6 alkylene, amino-substituted C1-C6 alkylene, hydroxy-substituted C1-C6 alkylene;

[0106] R 5 Selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -NH(C1-C6 alkyl), -O(C1-C6 alkyl), -N(C1-C6 alkyl)2, -NH(C3-C6 cycloalkyl) or the following groups:

[0107]

[0108] R 8 , R 10 Each independently selected from: a bond, a C1-C6 alkylene group;

[0109] R 9 Selected from: H, amino, hydroxyl, C1-C6 alkyl, C3-C6 cycloalkyl, -NHCH3, -N(CH3)2, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, -CF3, CHF2;

[0110] R 11 Selected from: C1-C6 alkyl, C3-C6 cycloalkyl, amino, hydroxyl, -CHF2, -CF3, piperazinyl, morpholinyl, piperidinyl, tetrahydropyranyl, tetrahydropyrrolyl, tetrahydrofuranyl, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2;

[0111] R 19 , R 20Each is independently selected from: H, C1-C6 alkyl; R 19 , R 20 It can also form a saturated C3-C6 heterocyclic group with the connected nitrogen atom, the heterocyclic group contains 1-2 heteroatoms, except for N, the other heteroatom is sulfur, oxygen or nitrogen; -(C1-C4 alkylene)-NH-(C1-C3 alkyl), -(C1-C4 alkylene)-N(C1-C3 alkyl)2;

[0112] R 23 It is H, C1-C6 alkyl.

[0113] More preferably, the compound has a structure shown in formula (VII)-(IX):

[0114]

[0115] Where R 1 Selected from: -CH2F, -CHF2, -CF3, C1-C6 alkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxy-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, -R 4 -R 5 Or a group like this:

[0116]

[0117] R a Selected from: isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio;

[0118] R e Selected from: isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, trifluoromethylthio, phenyl, pyridyl, 2-chloropyridin-4-yl, 2-cyanopyridin-4-yl, 2-methoxypyridin-4-yl;

[0119] R c Selected from: H, F, Cl, CN;

[0120] R 3’ Selected from: H, F, Cl, CN, -OR 8 R 9 , -R 10 -R 11 or -NR 19 R 20 ;

[0121] R 4 Selected from: C1-C6 alkylene;

[0122] R 5Selected from: -NH(C1-C6 alkyl), -O(C1-C6 alkyl), -N(C1-C6 alkyl)2 or the heterocyclic group shown below:

[0123]

[0124] R 8 , R 10 Each independently selected from: a bond, a C1-C6 alkylene group;

[0125] R 9 Selected from: H, amino, hydroxyl, C1-C6 alkyl, -NHCH3, -N(CH3)2, -CF3;

[0126] R 11 Selected from: C1-C6 alkyl, C3-C6 cycloalkyl, amino, hydroxy, -CF3, piperazinyl, morpholinyl, piperidinyl, tetrahydropyranyl, tetrahydropyrrolyl, tetrahydrofuranyl, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2;

[0127] R 19 , R 20 Each is independently selected from: H, C1-C6 alkyl; R 19 , R 20 It can also form a saturated C3-C6 heterocyclic group with the connected nitrogen atom, the heterocyclic group contains 1-2 heteroatoms, except for N, the other heteroatom is sulfur, oxygen or nitrogen; -(C1-C4 alkylene)-NH-(C1-C3 alkyl), -(C1-C4 alkylene)-N(C1-C3 alkyl)2;

[0128] R 23 It is H, C1-C3 alkyl.

[0129] Preferably, wherein R 1 Selected from: -CHF2, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxy-substituted C2-C4 alkyl, amino-substituted C2-C4 alkyl, -R 4 -R 5 or a heterocyclic group as shown below;

[0130]

[0131] R a Selected from: isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, trifluoromethylthio;

[0132] R eSelected from: isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, cyclobutyl, trifluoromethylthio, pyridyl, 2-chloropyridin-4-yl, 2-cyanopyridin-4-yl, 2-methoxypyridin-4-yl;

[0133] R c Selected from: H, F, Cl, CN;

[0134] R 3’ Selected from: H, F, Cl, CN, -OH, -OMe, -OCF3, -OCH2CH2NH2, -OCH2CH2NHMe, -OCH2CH2NMe2, -R 10 -R 11 or -NR 19 R 20 ;

[0135] R 4 Selected from: C2-C4 alkylene;

[0136] R 5 Selected from: -NHMe, -OMe, -NMe2 or the heterocyclic group shown below:

[0137]

[0138] R 10 Each independently selected from: a bond, a C1-C3 alkylene group;

[0139] R 11 Selected from: C1-C6 alkyl, C3-C6 cycloalkyl, amino, hydroxyl, -CF3, -NHMe, -NMe2;

[0140] R 19 , R 20 Each is independently selected from: H, C1-C3 alkyl, -CH2CH2NHMe, -CH2CH2NMe2; R 19 , R 20 It can also form a saturated C3-C6 heterocyclic group with the nitrogen atom to which it is attached. The heterocyclic group contains 1-2 heteroatoms, and in addition to N, the other heteroatom is sulfur, oxygen or nitrogen;

[0141] R 23 It is H, methyl, ethyl, isopropyl, and cyclopropyl.

[0142] Preferably, wherein R 1 Selected from: -CHF2, propyl, isopropyl, cyclopropyl, cyclobutyl, hydroxy-substituted C2-C4 alkyl, amino-substituted C2-C4 alkyl or the following groups:

[0143]

[0144] Ra Selected from: isopropyl, cyclopropyl;

[0145] R e Selected from: isopropyl, 1-(cyclopropyl)ethyl, cyclopropyl, trifluoromethylthio, 2-cyanopyridin-4-yl, 2-methoxypyridin-4-yl;

[0146] R c Selected from: H, F, Cl, CN;

[0147] R 3’ Selected from: H, F, Cl, CN, -OH, -Me, cyclopropyl, -OMe, -OCH2CH2NH2, -OCH2CH2NHMe, -OCH2CH2NMe2, -NMe2, -NHCH2CH2NMe2,

[0148]

[0149] Preferably, wherein R 1 Selected from: -CHF2, isopropyl, cyclopropyl, cyclobutyl or the following groups:

[0150]

[0151] R a is isopropyl;

[0152] R e Selected from: isopropyl, 1-(cyclopropyl)ethyl, trifluoromethylthio, 2-cyanopyridin-4-yl, 2-methoxypyridin-4-yl;

[0153] R c Selected from: H, F, Cl, CN;

[0154] R 3’ Selected from: H, Cl, CN, -Me, cyclopropyl, -OMe, -NMe2, -NHCH2CH2NMe2,

[0155] Preferably, wherein R 1 For isopropyl:

[0156] R a is isopropyl;

[0157] R e Selected from: isopropyl, 1-(cyclopropyl)ethyl, trifluoromethylthio, 2-cyanopyridin-4-yl, 2-methoxypyridin-4-yl;

[0158] R c Selected from: H, F, Cl, CN;

[0159] R 3’For H.

[0160] A second aspect of the present invention provides a compound selected from the group consisting of:

[0161]

[0162]

[0163] More preferably, the following compounds

[0164]

[0165] The third aspect of the present invention provides a pharmaceutical composition comprising the compound of the first or second aspect of the present invention or a pharmaceutically acceptable salt, solvate, prodrug and isomer thereof, and a pharmaceutically acceptable excipient.

[0166] The fourth aspect of the present invention provides a compound of the first or second aspect of the present invention or a pharmaceutically acceptable salt, solvate, prodrug and isomer thereof, or a pharmaceutical composition of the third aspect of the present invention, for use in medicine and / or for treating or preventing a disease, condition or illness. Typically, the use comprises administering the compound, salt, solvate, prodrug, isomer or pharmaceutical composition to a subject.

[0167] The fifth aspect of the present invention provides a compound of the first or second aspect of the present invention or a pharmaceutically acceptable salt, solvate, prodrug and isomer thereof, or a pharmaceutical composition of the third aspect of the present invention, for use in the manufacture of a medicament for treating or preventing a disease, condition or disorder.

[0168] The sixth aspect of the present invention provides a method for treating or preventing a disease, disorder or condition, the method comprising the step of administering an effective amount of a compound of the first or second aspect or a pharmaceutically acceptable salt, solvate, prodrug and isomer thereof, or a pharmaceutical composition of the third aspect of the present invention, thereby treating or preventing the disease, disorder or condition.

[0169] The seventh aspect of the present invention provides a compound of the first or second aspect of the present invention or a pharmaceutically acceptable salt, solvate, prodrug and isomer thereof, or a pharmaceutical composition of the third aspect of the present invention, for treating or preventing a disease, disorder or condition in an individual, wherein the individual has a germline or somatic non-silent mutation of NLRP3. The mutation may be, for example, a gain-of-function mutation or other mutation that causes increased NLRP3 activity.

[0170] The eighth aspect of the present invention provides a method for treating or preventing a disease, disorder or condition, the method comprising the following steps: diagnosing an individual with a germline or somatic non-silent mutation of NLRP3; and administering to the individual diagnosed positive an effective amount of a compound of the first or second aspect or a pharmaceutically acceptable salt, solvate, prodrug and isomer thereof, or a pharmaceutical composition of the third aspect of the present invention, thereby treating or preventing the disease, disorder or condition.

[0171] In general embodiments, the disease, disorder or condition can be a disease, disorder or condition of the immune system, cardiovascular system, endocrine system, gastrointestinal tract, renal system, liver system, metabolic system, respiratory system, central nervous system, can be cancer or other malignancy, and / or can be caused by or associated with a pathogen.

[0172] In one embodiment of the fifth, sixth, seventh, and eighth aspects of the invention, the disease, disorder, or condition is responsive to NLRP3 inhibition. As used herein, the term "NLRP3 inhibition" refers to a complete or partial reduction in the activity level of NLRP3, and includes, for example, inhibition of active NLRP3 and / or inhibition of NLRP3 activation.

[0173] Examples of diseases, disorders or conditions that may be responsive to NLRP3 inhibition and that can be treated or prevented according to the fifth, sixth, seventh and eighth aspects of the present invention include: inflammation, autoimmune diseases, infections, tumors, central nervous system diseases, cardiovascular diseases, metabolic diseases, respiratory diseases, liver diseases, kidney diseases, skin diseases, lymphatic vessel diseases, graft-versus-host disease, allodynia, and any disease in which an individual has been determined to carry a germline or somatic non-silent mutation in NLRP3.

[0174] In one embodiment of the fifth, sixth, seventh and eighth aspects of the invention, the disease, disorder or condition is: cold pyrin-associated periodic syndrome (CAPS), Mueller-Weiss syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), neonatal onset multisystem inflammatory disease (NOMID), chronic infantile neurocutaneous and articular syndrome, familial Mediterranean fever (FMF), pyoderma gangrenosum and acne syndrome (PAPA), hyperimmunoglobulinemia D and periodic fever syndrome (HIDS), tumor necrosis factor (TNF) receptor-associated periodic syndrome (TRAPS), adult-onset Still's disease (AOSD), relapsing polychondritis, Schnitzler syndrome, Sweet syndrome, Behcet's disease, antisynthetase syndrome, interleukin-1 receptor antagonist deficiency (DIRA), tumors, type 2 diabetes, obesity, gout, atherosclerosis, rheumatoid arthritis, multiple sclerosis, amyotrophic lateral sclerosis, systemic lupus erythematosus, inflammatory bowel disease, hepatitis, nonalcoholic steatohepatitis, alcoholic liver disease, kidney disease, silicosis, silicosis, asbestosis, asthma, acute respiratory distress syndrome, depression, Parkinson's disease, Alzheimer's disease, aging, contact hypersensitivity, blindness caused by degeneration of visual pigment cells, sepsis.

[0175] Example - Compound Synthesis

[0176] Unless otherwise stated, all reagents, solvents and compounds described below were purchased from commercial analytical or chemically pure reagents and used directly without further purification.

[0177] abbreviation

[0178] AcCl Acetyl chloride

[0179] Ac2O Acetic anhydride

[0180] AcOH Acetic acid

[0181] Bn Benzyl

[0182] aq aqueous solution

[0183] DCM Dichloromethane

[0184] DIPEA N,N-Diisopropylethylamine

[0185] DMSO Dimethyl sulfoxide

[0186] DMF N,N-Dimethylformamide

[0187] equiv equivalent

[0188] EA Ethyl acetate

[0189] ESI Electrospray ionization

[0190] Et Ethyl

[0191] EtOH

[0192] FA Formic acid

[0193] h hour

[0194] HPLC High Performance Liquid Chromatography

[0195] LC Liquid Chromatography

[0196] LDA Lithium diisopropylamide

[0197] Me Methyl

[0198] MeCN Acetonitrile

[0199] MeOH Methanol

[0200] [M+H] + Protonated molecular ion

[0201] MHz Megahertz

[0202] min

[0203] MS

[0204] m / z mass-to-charge ratio

[0205] NCS N-Chlorosuccinimide

[0206] NIS N-iodosuccinimide

[0207] NMR Nuclear Magnetic Resonance (Spectroscopy)

[0208] Pd2(dba)3 tris(dibenzylideneacetone)dipalladium

[0209] Pd(dppf)Cl2 [1,1'-Bis(diphenylphosphino)ferrocene]palladium dichloride

[0210] PE Petroleum Ether

[0211] Phen 1,10-Phenanthroline

[0212] prep-HPLC Preparative High Performance Liquid Chromatography

[0213] rt room temperature

[0214] t BuOK Potassium tert-butoxide

[0215] TEA Triethylamine

[0216] TFAA trifluoroacetic acid

[0217] THF Tetrahydrofuran

[0218] v / v volume ratio

[0219] wt% Weight % or percentage by weight

[0220] Xantphos 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene

[0221] Experimental methods

[0222] Thin layer analysis and column chromatography:

[0223] The thin layer analysis (TLC) plate model is HSGF-254 (thickness 0.15-0.2 mm, produced by Yantai Chemical Experimental Plant); the column chromatography silica gel is 200-300 mesh commercial silica gel produced by Qingdao Ocean Chemical Plant.

[0224] NMR:

[0225] 1 H-NMR spectra were recorded at 200 MHz, 400 MHz or 500 MHz. Unless otherwise stated, spectra were measured at 298 K. 1 H-NMR was recorded using a Varian Mercury-300, Varian Mercury-400 or Varian Mercury-500 NMR instrument with tetramethylsilane (TMS) as the internal standard; chemical shifts were (ppm, δ), and proton coupling labels were singlet (s), doublet (d), triplet (t), quartet (q), multiplet (m), broad (br), and septet (hept).

[0226] LC-MS:

[0227] Instruments used: Agilent 1260II-6120B, SHIMADZU LCMS-2020; General method: Column model YMCTriart C18, 50*4.6mm5μm; Mobile phase A: water (0.1% formic acid), B: acetonitrile; Flow rate 2.5mL / min, over 2.9 minutes, gradient elution with aqueous solution containing 0.1% v / v formic acid and acetonitrile, UV detection at 220nm and 254nm. Gradient information: 0.0-0.1 minutes, 20% acetonitrile; 0.1-1.8 minutes, from 20% acetonitrile to 95% acetonitrile; 1.8-2.5 minutes, maintained at 95% acetonitrile; 2.5-2.9 minutes, 20% acetonitrile.

[0228] Prep-HPLC:

[0229] Column model: YMC Triart C18, 250*20 mm ID 5 μm; mobile phase A: water (0.1% NH 3 H 2 O), B: acetonitrile. DETAILED DESCRIPTION

[0230] In order to illustrate the present invention in more detail, the following examples are given, but the scope of the present invention is not limited thereto.

[0231] Synthetic intermediates

[0232] Intermediate L1: 1-isopropyl-1H-pyrazolo[4,3-b]pyridine-3-sulfonamide

[0233]

[0234] Step 1: 3-Bromo-1H-pyrazolo[4,3-b]pyridine (10 g, 50.5 mmol, 1.00 equiv.) was dissolved in 250 mL of N,N-dimethylformamide, and sodium hydride (2.2 g, 55.6 mmol, 60%, 1.10 equiv.) was added at 0°C under nitrogen protection. After the reaction mixture was stirred at 0°C for 30 minutes, 2-iodopropane (9.44 g, 55.6 mmol, 1.10 equiv.) was added at 0°C. The reaction mixture was stirred at room temperature for 3 hours, and then the reaction system was quenched with NH4Cl (aq.). Ethyl acetate was added to the solution, and the organic phase was washed with water and saturated sodium chloride solution successively, and dried over anhydrous sodium sulfate. The organic phase was concentrated in vacuo, and the crude product was separated and purified by silica gel column chromatography to obtain S1 (11.9 g, 98%).

[0235] 1 H NMR (400MHz, DMSO-d6): δ8.60(dd,J=4.3,1.2Hz,1H),8.30(dd,J=8.6,0.9Hz,1H),7.51(m,1H),5.06(m,1H),1.49(d,J=6.6Hz,6H).

[0236] LRMS (ESI) m / z: 240.0 [M+H] +

[0237] Step 2: S1 (6 g, 25.0 mmol, 1.00 equiv.) was dissolved in 250 mL of 1,4-dioxane, and benzyl mercaptan (4.65 g, 37.5 mmol, 1.50 equiv.), tris(dibenzylideneacetone)dipalladium (1.1 g, 1.3 mmol, 0.05 equiv.), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (1.4 g, 2.5 mmol, 0.10 equiv.) and N,N-diisopropylethylamine (6.45 g, 50.0 mmol, 2.00 equiv.) were added successively under nitrogen protection. The reaction system was stirred at 100 ° C for 16 h, cooled to room temperature, and the mixture was filtered through celite, and the filter cake was washed with ethyl acetate. The solution was diluted with ethyl acetate, and the organic phase was washed with water and saturated sodium chloride solution successively, and dried over anhydrous sodium sulfate. The organic phase was concentrated in vacuo and the crude product was purified by silica gel column chromatography to give S2 (7.0 g, 99%).

[0238] 1 H NMR (400MHz, DMSO-d6): δ8.51(dd,J=4.3,1.3Hz,1H),8.19(dd,J=8.6,1.3Hz,1H),7.42(dd,J=8.6,4.3Hz,1H ),7.33–7.29(m,2H),7.28–7.22(m,2H),7.20(m,1H),5.00–4.95(m,1H),4.47(s,2H),1.46(d,J=6.6Hz,6H).

[0239] LRMS (ESI) m / z: 284.1 [M+H] +

[0240] Step 3: S2 (10.8 g, 38.0 mol, 1.0 equiv.) was dissolved in a mixed solvent of acetic acid (156 mL) and water (52 mL), and N-chlorosuccinimide (15.12 g, 114.0 mmol, 3.0 equiv.) was added under ice bath, and then reacted for 6 h at room temperature under nitrogen protection. After the reaction was completed, 200 mL of ethyl acetate was added for dilution, and the organic phase was washed successively with water (200 mL*2) and saturated sodium chloride solution (60 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to obtain the crude product S3, which was directly used in the next step.

[0241] Step 4: S3 (10.8 g, 38.0 mol, 1.0 equiv.) was dissolved in 54 mL of acetone, and a solution of NH4HCO3 (59.4 g, 623.2 mmol, 16.4 equiv.) dissolved in 162 mL of water was added at 0°C under nitrogen protection. The reaction system was stirred at room temperature overnight under nitrogen protection. After the reaction was completed, 200 mL of ethyl acetate was added to the reaction solution, and the organic phase was washed with water (200 mL*2) and then with 150 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo and purified by silica gel column chromatography (PE:EA=30:70) to obtain intermediate L1 (1.4 g, 14%).

[0242] 1 H NMR (400MHz, DMSO-d6): δ8.70 (dd, J=4.3, 1.3Hz, 1H), 8.38 (dd, J=8.7, 1.3Hz, 1H),7.68(s,2H),7.56–7.51(m,1H),5.21–5.08(m,1H),1.53(d,J=6.6Hz,6H).

[0243] LRMS (ESI) m / z: 241.1 [M+H] +

[0244] The following intermediates were prepared according to the general synthetic method of intermediate L1:

[0245]

[0246] Intermediate L6: 6-dimethylamino-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-3-sulfonamide

[0247]

[0248] Step 1: At -78°C, add 2M lithium diisopropylamide tetrahydrofuran solution (65.2mL) to a solution of 2,6-difluoropyridine (15.0g, 130.3mmol) in tetrahydrofuran (500mL). Stir at -78°C for 1h, then add N,N-dimethylformamide (20.1mL, 260.7mmol). After the addition is complete, the reaction system continues to stir at -78°C for 2h, and the reaction is completed by LC-MS monitoring. The reaction system is quenched with saturated NH4Cl solution and extracted with dichloromethane (500mL×3). The organic layer is washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and the filtrate is concentrated, and purified by silica gel column chromatography (PE:EA=20:1) to obtain yellow solid S4 (17.0g, 78%).

[0249] LRMS (ESI) m / z: 169.1 [M+H]+

[0250] Step 2: S4 was dissolved in 120 mL of ethanol, and then hydrazine hydrate (85%, 60 mL) was added under stirring at room temperature, and then the reaction system was heated to 80°C and stirred overnight. After the reaction was completed as monitored by LC-MS, the reaction system was concentrated under reduced pressure to remove most of the ethanol, the residue was filtered, the filter cake was washed with cold methanol, and vacuum dried to obtain a white solid crude product S5 (14.0 g, 85%), which was directly used in the next step.

[0251] LRMS (ESI) m / z: 163.2 [M+H] +

[0252] Step 3: S5 (14.0 g, 86.32 mmol) was added to 1 L of acetonitrile, and then N-iodosuccinimide (23.3 g, 103.58 mmol) was added, and the mixture was stirred at 50°C overnight. After the reaction was completed by LC-MS monitoring, the reaction system was quenched with a saturated Na2S2O3 solution, and then extracted with dichloromethane (500 mL×3). The organic phase was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated, and purified by silica gel column chromatography (DCM: MeOH = 100: 1) to obtain a light yellow solid S6 (12.0 g, 48%).

[0253] 1 H NMR (400MHz, CDCl3): δ7.83 (s, 1H), 7.80 (d, J = 9.0Hz, 1H), 6.52 (d, J = 9.0Hz, 1H), 3.25 (s, 6H).

[0254] LRMS (ESI) m / z: 289.0 [M+H] +

[0255] Step 4: Add 12.0 g of S6 to 200 mL of N,N-dimethylformamide, then add 2-iodopropane (35.4 g, 208.3 mmol) and cesium carbonate (13.6 g, 41.7 mmol), and stir the reaction mixture at 30 ° C overnight. After the reaction is completed by LC-MS monitoring, add 300 mL of water to dilute, and extract with ethyl acetate (300 mL×2). The organic phase is washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. Filter and concentrate the filtrate, and purify it by silica gel column chromatography (PE:EA=9:1) to obtain a white solid S7 (7.2 g, 52%).

[0256] LRMS (ESI) m / z: 331.0 [M+H] +

[0257] Step 5: S7 (7.1 g, 21.5 mmol) and benzyl mercaptan (3.5 g, 28.0 mmol) were added to 150 mL of dioxane, and then tris(dibenzylideneacetone)dipalladium (1.97 g, 2.15 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (2.49 g, 4.30 mmol) and N,N-diisopropylethylamine (7.7 mL, 43.0 mmol) were added successively. The reaction was carried out at 100°C for 16 h. After the reaction was completed as monitored by LC-MS, 300 mL of dichloromethane was added to dilute the reaction mixture, and the organic phase was washed successively with water and saturated sodium chloride solution, and then dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated, and purified by silica gel column chromatography (PE:EA=5:1) to obtain white solid S8 (7 g, 98%).

[0258] LRMS (ESI) m / z: 327.2 [M+H] +

[0259] Step 6: At 0°C, S8 (7.0 g, 21.5 mmol) was added to a mixed solvent of 210 mL of acetic acid and 70 mL of water, and N-chlorosuccinimide (8.57 g, 64.4 mmol) was added in batches while stirring. The reaction was carried out at room temperature for 16 h. After the reaction was completed as monitored by LC-MS, 500 mL of water was added to dilute the reaction system, and ethyl acetate (200 mL×3) was used for extraction. The organic phase was washed with saturated NaHCO3 solution and saturated sodium chloride solution, and then dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated, and purified by silica gel column chromatography (PE:EA=9:1) to obtain white solid S9 (4.88 g, 75%).

[0260] Step 7: Add an aqueous solution of ammonia (25% wt., 5 mL) dropwise to a 50 mL solution of S9 (4.88 g, 16.2 mmol) in dichloromethane. The reaction system was stirred at room temperature for 2 h. After LC-MS monitoring of the reaction, 150 mL of water was added to dilute the reaction mixture and extracted with dichloromethane (80 mL × 3). The organic phase was washed with a saturated sodium chloride solution and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated to obtain a white solid L6 (3.75 g, 82%).

[0261] 1 H NMR (400MHz, DMSO-d6): δ7.99(d,J=9.1Hz,1H),7.60(s,2H),6.82(d,J=9.1Hz,1H),5.05(m,1H),3.15(s,6H),1.49(d,J=6.7Hz,6H).

[0262] LRMS (ESI) m / z: 284.4 [M+H] +

[0263] Intermediate R1: 8-fluoro-1,2,3,5,6,7-hexahydro-s-indacene-4-amine

[0264]

[0265] Step 1: 1,2,3,5,6,7-hexahydro-s-indacen-4-amine (18 g, 104.0 mmol, 1.0 equiv.) and triethylamine (12.6 g, 125 mmol, 1.2 equiv.) were dissolved in 200 mL of anhydrous dichloromethane, acetic anhydride (11.64 g, 114.1 mmol) was added at 0°C, and the mixture was stirred at 25°C for 16 h. Subsequently, 200 mL of water was added to the reaction system, filtered, the solid was washed with water, and vacuum dried to obtain the crude product S10 (20 g, 89%), which was directly used in the next step.

[0266] LRMS (ESI) m / z: 216.2 [M+H] +

[0267] Step 2: S10 (2 g, 9.3 mmol, 1.0 equiv.) was dissolved in 30 mL of dichloromethane, and pyridine hydrofluoride solution (10 mL, 111.6 mmol, 12 equiv.) was added at 0°C under nitrogen protection, and [bis(trifluoroacetoxy)iodo]benzene (6 g, 14.0 mmol, 1.5 equiv.) was added, and the reaction mixture was stirred at 0°C for 1 h. After the reaction was completed, sodium hydroxide (aq, 4 M) solution was added at 0°C, and the precipitated solid was filtered out. The organic phase was washed with water and saturated sodium chloride solution successively, and then dried over anhydrous sodium sulfate. After filtration and vacuum concentration, S11 (500 mg, 23%) was purified by silica gel column chromatography.

[0268] LRMS (ESI) m / z: 234.1 [M+H] +

[0269] Step 3: S11 (4 g, 17.1 mmol, 1.0 equiv.) and 12 M HCl (10 mL) were added successively to 100 mL of ethanol, and the reaction system was heated to reflux under argon protection for 16 h. The reaction mixture was then cooled to room temperature and the organic solvent was concentrated under reduced pressure. The concentrate was neutralized with 2 M aqueous sodium hydroxide solution, transferred to a separation funnel, and extracted with dichloromethane (100 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo, and then purified by silica gel column chromatography to obtain intermediate R1 (1.7 g, 52%).

[0270] 1H NMR (400MHz, CDCl3): δ3.33 (s, 2H), 2.90 (t, J = 7.4Hz, 4H), 2.71 (t, J = 7.3Hz, 4H), 2.19-2.10 (m, 4H). 19 F NMR (377MHz, CDCl3): δ-135.01 (s).

[0271] LRMS (ESI) m / z: 192.3 [M+H] +

[0272] Intermediate R2: 8-chloro-1,2,3,5,6,7-hexahydro-s-indacene-4-amine

[0273]

[0274] Steps: 1,2,3,5,6,7-hexahydro-s-indacen-4-amine (350 mg, 2.02 mmol, 1.00 equiv.) was dissolved in 10 mL of acetonitrile, and N-chlorosuccinimide (287 mg, 2.15 mmol, 1.05 equiv.) was added. After stirring at room temperature for 18 h, the reaction system was spin-dried, the solid produced was washed with water, the filter cake was collected after filtration and vacuum dried to obtain a white solid intermediate R2 (202 mg, 90%).

[0275] 1 H NMR (400MHz, CDCl3): δ4.68 (s, 1H), 2.92 (t, J = 7.4Hz, 4H), 2.82 (t, J = 7.4Hz, 4H), 2.18–2.09 (m, 4H).

[0276] LRMS (ESI) m / z: 208.3 [M+H] +

[0277] Intermediate R3: 8-cyano-1,2,3,5,6,7-hexahydro-s-indacene-4-amine

[0278]

[0279] Step 1: 1,2,3,5,6,7-hexahydro-s-indacene-4-amine (1.73 g, 10 mmol, 1.0 equiv) was dissolved in 20 mL of N,N-dimethylformamide, and then N-iodosuccinimide (2.48 g, 11 mmol, 1.1 equiv) was added. The reaction system was reacted at room temperature overnight, and then 100 mL of ethyl acetate was added to the reaction solution. The organic phase was washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and dried by spin drying, and then purified by silica gel column chromatography (PE:EA=25:1) to obtain S12 (2.54 g, 85%).

[0280] LRMS (ESI) m / z: 300.0 [M+H] +

[0281] Step 2: The intermediate S12 (1.50 g, 5.0 mmol, 1.0 equiv.) was dissolved in 10 mL of N,N-dimethylformamide, and potassium tert-butoxide (561 mg, 5 mmol, 1.0 equiv.), cuprous cyanide (672 mg, 7.5 mmol, 1.5 equiv.) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (733 mg, 1 mmol, 0.2 equiv.) were added successively under nitrogen protection. The reaction system was stirred at 120°C for 12 h, then cooled to room temperature, 60 mL of ethyl acetate was added to the reaction solution, the organic phase was washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, spin-dried, and purified by silica gel column chromatography (PE:EA=20:1) to obtain the intermediate R3 (575 mg, 58%).

[0282] 1 H NMR (500MHz, DMSO-d6) δ5.64(s,2H),2.84(t,J=7.5Hz,4H),2.63(t,J=7.3Hz,4H),2.10–1.96(m,4H).

[0283] LRMS (ESI) m / z: 199.3 [M+H] +

[0284] Intermediate R4: 5-(1-cyclopropylethyl)-2,3-dihydro-1H-indene-4-amine

[0285]

[0286] Step 1: Dissolve the substrate 2,3-dihydro-1H-inden-4-amine (1.33 g, 10 mmol, 1.0 equiv.) and cyclopropanenitrile (670 mg, 10 mmol, 1.0 equiv.) in 25 mL of anhydrous dichloromethane, add aluminum chloride (133 mg, 1 mmol, 0.1 equiv.), protect with nitrogen, slowly dropwise add 1N boron trichloride dichloromethane solution (10 mL, 10 mmol, 1.0 equiv.) at 0°C, then slowly warm to room temperature to react overnight. After the reaction is completed by TLC monitoring, pour the reactant into ice water, separate the liquids with a separatory funnel, wash the organic phase with a saturated sodium chloride solution, dry with anhydrous sodium sulfate, and then concentrate in vacuo to obtain the crude product S13 for use.

[0287] LRMS (ESI) m / z: 202.1 [M+H] +

[0288] Step 2: Dissolve the crude product S13 in 25 mL of anhydrous tetrahydrofuran, protect with nitrogen, slowly dropwise add 1N methylmagnesium bromide Grignard reagent tetrahydrofuran solution (10 mL, 10 mmol, 1.0 equiv.) at 0°C, then slowly warm to room temperature and react overnight. After the reaction is completed by TLC monitoring, pour the reactant into an ice-cold saturated ammonium chloride solution, extract with ethyl acetate (3*50 mL), wash the organic phase with a saturated sodium chloride solution, dry with anhydrous sodium sulfate, and then concentrate in vacuo to obtain the crude product S14 for use.

[0289] LRMS (ESI) m / z: 218.2 [M+H] +

[0290] Step 3: The crude product S14 was dissolved in 50 mL of anhydrous dichloromethane, triethylsilane (5.8 g, 50 mmol, 5.0 equiv.) was added, and trifluoroacetic acid (3.42 g, 30 mmol, 5.0 equiv.) was slowly added dropwise at 0°C under nitrogen protection, and then the temperature was slowly raised to room temperature and stirred for 12 hours. The reaction mixture was poured into an ice-cold saturated sodium bicarbonate solution and extracted with ethyl acetate (3*50 mL), and the combined organic phases were washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo, and purified by silica gel column chromatography (PE:EA=10:1) to obtain a light brown oil R4 (848 mg, 42%).

[0291] 1H NMR (600MHz, DMSO-d6) δ6.92(d,J=7.6Hz,1H),6.45(d,J=7.6Hz,1H),4.43(s,2H),2.74(t,J=7.4Hz,2H),2.68–2.56(m,2H),2. 23(m,1H),1.95(q,J=7.4Hz,2H),1.14(d,J=6.8Hz,3H),1.04–0.94(m,1H),0.47(m,1H),0.31(m,1H),0.13(m,1H),0.02(m,1H).

[0292] LRMS (ESI) m / z: 202.2 [M+H] +

[0293] Intermediate R5: 7-chloro-5-trifluoromethylthio-2,3-dihydro-1H-indene-4-amine

[0294]

[0295] Step 1: 2,3-dihydro-1H-inden-4-amine (1 g, 7.5 mmol, 1.0 equiv.) was dissolved in 30 mL of anhydrous ethanol and cooled to 0°C. Acetic anhydride (1.9 g, 18.74 mmol, 2.5 equiv.) was added dropwise under nitrogen protection. The resulting reaction mixture was warmed to room temperature and stirred for 3 hours. After the reaction was completed by TLC monitoring, the reaction mixture was concentrated in vacuo, and the resulting residue was diluted with ether, filtered and dried in vacuo to obtain intermediate S15 as a white solid (1.2 g, 90%).

[0296] 1 HNMR(400MHz,DMSO-d6)δ9.29(s,1H),7.41(d,J=8.0Hz,1H),7.08(t,J=7.6Hz,1H),6.99(d, J=6.8Hz,1H),2.87(t,J=7.2Hz,2H),2.80(t,J=7.2Hz,2H),2.04(s,3H),1.99-1.95(m,2H).

[0297] LRMS (ESI) m / z: 176.4 [M+H] +

[0298] Step 2: S15 (1 g, 5.55 mmol, 1.0 equiv.) was dissolved in 10 mL of acetic acid and cooled to 0 °C, N-chlorosuccinimide (1.1 g, 8.45 mmol, 1.5 equiv.) was added in portions, and the mixture was warmed to room temperature and stirred overnight. After the reaction was completed by TLC monitoring, the reactant was poured into ice water, filtered, washed with saturated NaHCO3 solution, saturated Na2S2O3 solution and dried in vacuo to obtain a white solid S16 (0.6 g, 50%). 1 HNMR (300MHz, CDCl3) δ7.73(d,J=8.4Hz,1H),7.14(d,J=8.4Hz,1H),6.86(s,lH),3.02-2.85(m,4H),2.19(s,3H),1.99(m,2H).

[0299] LRMS (ESI) m / z: 209.8 [M+H] +

[0300] Step 3: S16 (600 mg, 2.85 mmol, 1.0 equiv.) was dissolved in 25 mL of 3N HCl solution and heated to 90 °C for 4 hours. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature, poured into saturated NaHCO3 solution, and extracted with ethyl acetate (3*30 mL). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated in vacuo to obtain white solid S17 (350 mg, 74%).

[0301] 1 HNMR (300M Hz, DMSO-d6) δ6.85(d,J=8.4Hz,1H),6.40(d,J=8.4Hz,1H),4.97(s,2H),2.82(t,J=8.1Hz,2H),2.71(t,J=7.5Hz,2H),2.01-1.96(m,2H).

[0302] LRMS (ESI) m / z: 168.2 [M+H] +

[0303] Step 4: S17 (1.67 g, 10 mmol, 1.0 equiv.) was dissolved in acetonitrile, N-iodosuccinimide (2.25 g, 10 mmol, 1.0 equiv.) was slowly added in batches at 0°C, and then the temperature was slowly raised to room temperature and stirred for 12 hours. After the reaction was completed by TLC monitoring, the reaction mixture was diluted with saturated Na2S2O3 solution and extracted with ethyl acetate (3*50 mL). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated in vacuo. Purification by silica gel column chromatography (PE:EA=10:1) gave a brown-red solid S18 (880 mg, 30%).

[0304] 1 HNMR(300M Hz,DMSO-d6)δ7.36(s,1H),5.04(s,2H),2.82-2.72(m,4H),2.03-1.98(m,2H).LRMS(ESI)m / z:293.7[M+H] +

[0305] Step 5: S18 (2.93 g, 10 mmol, 1.0 equiv.) was dissolved in 25 mL of dichloromethane, triethylamine (2.02 g, 20 mmol, 2.0 equiv.) was added, acetyl chloride (785 mg, 10 mmol, 1.0 equiv.) was slowly added dropwise at 0°C, and then the temperature was slowly raised to room temperature for 4 hours. After the reaction was completed by TLC monitoring, the reaction mixture was diluted with dichloromethane and washed with water (3*50 mL). The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated in vacuo to obtain crude S19 for use.

[0306] LRMS (ESI) m / z: 336.0 [M+H] +

[0307] Step 6: The product S19 of the previous step was completely dissolved in 25 mL of acetonitrile, and silver trifluoromethylsulfide (2.57 g, 10 mmol, 1.0 equiv.), cuprous bromide (143 mg, 1 mmol, 0.1 equiv.), and 1,10-phenanthroline (180 mg, 1 mmol, 0.1 equiv.) were added, and the mixture was heated to 80 ° C under nitrogen protection for 12 hours. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature, poured into a saturated NaHCO3 solution, and extracted with ethyl acetate (3*50 mL). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated in vacuo, and purified by silica gel column chromatography (PE:EA=4:1) to obtain a white solid S20 (1.92 g, 62%).

[0308] 1HNMR (300M Hz, DMSO-d6) δ7.25(s,lH),6.24(s,1H),2.82-2.78(m,4H),2.01-1.95(m,2H),2.04(s,3H).

[0309] LRMS (ESI) m / z: 310.0 [M+H] +

[0310] Step 7: S20 (1 g, 3.23 mmol) was dissolved in 40 mL of 3N HCl solution, and the reaction mixture was prepared according to step 3 to obtain R5 (786 mg, 91%) as a white solid.

[0311] 1 HNMR(300M Hz,DMSO-d6)δ7.01(s,lH),5.35(s,2H),2.85-2.81(m,4H),2.03-1.99(m,2H).LRMS(ESI)m / z:268.0[M+H] +

[0312] Preparation Example

[0313] Embodiment 1:

[0314] N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-1-isopropyl-1H-pyrazolo[4,3-b]pyridine-3-sulfonamide

[0315]

[0316] Step 1: At 0°C, triphosgene (24 mg, 0.08 mmol) and triethylamine (20 mg, 0.2 mmol) were dissolved in 3 mL of tetrahydrofuran solution, and 1,2,3,5,6,7-hexahydro-s-indacen-4-amine (17.3 mg, 0.1 mmol) was added. The mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction system was concentrated to obtain a crude product 1,2,3,5,6,7-hexahydro-s-indacen-4-isocyanate (20.1 mg, 100%), which was directly used in the next step.

[0317] Step 2: At 0°C, sodium hydride (8 mg, 0.20 mmol, 60%, dispersed in mineral oil) was added to a solution of L1 (24 mg, 0.1 mmol) in THF (3 mL). After stirring at 0°C for 10 min, a solution of 1,2,3,5,6,7-hexahydro-s-indacene-4-isocyanate (20.1 mg, 0.1 mmol) in tetrahydrofuran (2 mL) was added. Stir at room temperature for 2 h. After the reaction was completed as monitored by LC-MS, water was added to quench the mixture. The mixture was extracted with ethyl acetate, and the organic phase was washed with water and saturated sodium chloride solution successively, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The mixture was purified by prep-HPLC (C18, Phase A: water (0.5% NH3·H2O), Phase B: methanol) to obtain 1 (20.5 mg, 46%) as a white solid.

[0318] 1 H NMR (400MHz, DMSO-d6): δ8.55(d,J=4.2Hz,1H),8.23(d,J=8.5Hz,1H),7.65(s,1H),7.48-7.39(m,1H),7.19(s,1H) ,6.77(s,1H),5.05(m,1H),2.72(t,J=7.3Hz,4H),2.57(t,J=7.2Hz,4H),1.90-1.81(m,4H),1.49(d,J=6.6Hz,6H).

[0319] LRMS (ESI) m / z: 440.1 [M+H] +

[0320] The following compounds were prepared according to the general synthetic method of compound 1:

[0321]

[0322]

[0323]

[0324] Example - Biological Research

[0325] NLRP3 and pyroptosis

[0326] The NLRP3 inflammasome of macrophages can be activated by the first signal such as LPS and the second signal such as ATP. Under the action of the activated NLRP3 inflammasome, proinflammatory cytokines such as IL-1β mature and are released outside the cell. Therefore, the amount of IL-1β in the cell culture supernatant can reflect the activity and inhibition of the NLRP3 inflammasome.

[0327] BMDM cell preparation

[0328] 1. In a clean bench, remove the hind leg bones of C57BL / 6 mice (purchased from Shanghai Slake Experimental Animal Co., Ltd.), strip the muscle tissue, keep the femur and tibia connected and intact, wash twice in RPMI.1640 medium (Senrui Biological, CR31800) containing 20% ​​fetal bovine serum (Gemini, 10099-141-C), separate the femur and tibia at the knee joint with scissors, cut the two ends of the bone, blow out the bone marrow with a 1mL syringe, and blow the syringe several times to disperse the bone marrow. After passing the cell suspension through a 200-mesh sieve, centrifuge it at 4°C, 1000rpm for 5 minutes. Discard the supernatant and collect the bone marrow cells;

[0329] 2. Add 1 mL of 1× red blood cell lysis buffer (Boster, AR1118) to the bone marrow cells, incubate on ice for 4 min, centrifuge at 1000 rpm for 5 min, discard the red supernatant, and wash twice with RPMI.1640 medium;

[0330] 3. Bone marrow cells were induced to differentiate in RPMI.1640 medium containing 20 ng / mL M-CSF (R&D, 216-MC / CF). The medium was changed on the 3rd day and mature BMDMs were obtained after 5 days of induction.

[0331] Pyroptosis assay of BMDM

[0332] 1. Digest the adherent macrophages with trypsin, collect and plate on a 96-well plate, with 5×10 cells per well. 4 After overnight culture, 100 μL of 20 ng / mL LPS (Sigma, L2630-10MG) was added to each well, with a final concentration of 10 ng / mL, and incubated for 3 h. The cells were then divided into 7 groups, and the settings of each group are shown in Table 1:

[0333] Table 1

[0334]

[0335] Group 1 was the blank control group: LPS was removed, 200 μL RPMI.1640 medium was added, and incubated for 0.5 h, then the medium was changed and RPMI.1640 medium was added, 200 μL per well;

[0336] The second group was the solvent control group: LPS was removed, 0.1% DMSO (v / v) was added and incubated for 0.5 h, then the solution was changed and 5 mM ATP (Mei Lun Bio, MB3157) was added, 200 μL per well;

[0337] Groups 3 to 7 were added with compounds 1-12 obtained in the above-mentioned preparation examples or positive control drugs MCC950 / MCC7840: LPS was removed, and 200 μL of different concentrations of compounds 1-12 or positive control drugs MCC950 / MCC7840 were added, with concentrations of 2, 0.4, 0.08, 0.016 and 0.0032 μM (obtained by diluting the mother solutions dissolved in 2, 0.4, 0.08, 0.016 and 0.0032 mM DMSO with RPMI.1640 medium, respectively), incubated for 0.5 h, and then the medium was changed, and 5 mM ATP was added, 200 μL per well;

[0338] 2. After incubation for 1 hour, collect the cell culture supernatant and detect IL-1β in the supernatant using an ELISA kit (R&D, DY401-15). Determine the absorbance of each group at 450nm, analyze the data using Graph Pad software, and calculate the IC50 value.

[0339] The results of the pyroptosis assay are summarized in Table 2 below as BMDM IC50.

[0340] Table 2

[0341]

[0342]

[0343] *: Data from the literature: Jiang H et al., J Exp Med. 2017 Nov 6; 214(11): 3219-3238

[0344] #: Data from the literature: He Y et al., J Biol Chem. 2014 Jan 10; 289(2): 1142-50

[0345] As shown in the results in the above table, the compounds obtained in Examples 1-12 all showed certain inhibitory activity against NLRP3 inflammasome, especially the compounds obtained in Examples 1, 3, 5, 7-10 had better inhibitory activity.

Claims

1. A compound represented by formula (II)-(VI), or a pharmaceutically acceptable salt thereof: Where R 1 is a C1-C6 alkyl group; R 2 Selected from: in B 1 and B 2 is -CH2CH2CH2-; R a , R e Selected from C1-C6 alkyl; R c Selected from: H, halogen, cyano; R b , R d is H; R f Selected from C3-C6 cycloalkyl substituted C1-C6 alkyl, -S(C1-C6 alkyl), trifluoromethylthio; R 3’ Selected from: H, -NMe2.

2. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound has a structure represented by formula (III): Where R 1 is a C1-C6 alkyl group; R 2 Selected from: in B 1 and B 2 is: -CH2CH2CH2-; R a isopropyl; R e isopropyl; R f Selected from 1-(cyclopropyl)ethyl, trifluoromethylthio; R b , R d For: H; R c Selected from: H, F, Cl, CN; R 3’ Selected from: H, -NMe2.

3. The compound or pharmaceutically acceptable salt thereof according to claim 2, wherein the compound has a structure shown in formula (VII)-(IX): Where R 1 Selected from C1-C6 alkyl; R a Selected from: isopropyl; R e Selected from: isopropyl; R c Selected from: H, F, Cl, CN; R 3’ Selected from: H, or -NMe2.

4. The compound or pharmaceutically acceptable salt thereof according to claim 3, wherein the compound has a structure shown in formula (VII)-(IX): Where R 1 is isopropyl; R a Selected from: isopropyl; R e Selected from: isopropyl; R c Selected from: H, F, Cl, CN; R 3’ Selected from: H, -NMe2.

5. The compound or pharmaceutically acceptable salt thereof according to claim 4, wherein the compound has a structure shown in formula (VII)-(IX): Where R 1 is isopropyl; R a is isopropyl; R e is isopropyl; R c Selected from: H, F, Cl, CN; R 3’ For H.

6. The compound described below or a pharmaceutically acceptable salt thereof:

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

8. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

9. Use of a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 8 in the preparation of a medicament for treating or preventing a disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

10. The use according to claim 9, wherein the disease, disorder or condition is selected from inflammation, autoimmune disease, infection, tumor, central nervous system disease, cardiovascular disease, metabolic disease, respiratory disease, liver disease, kidney disease, skin disease, lymphatic vessel disease, graft-versus-host disease, allodynia and any disease in which an individual has been determined to carry a germline or somatic non-silent mutation of NLRP3.

11. The use of claim 9, wherein the disease, disorder or condition is selected from the group consisting of cold pyrin-associated periodic syndrome (CAPS), Mueller-Weiss syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), neonatal-onset multisystem inflammatory disease (NOMID), chronic infantile neurocutaneous and articular syndrome, familial Mediterranean fever (FMF), pyoderma gangrenosum and acne syndrome (PAPA), hyperimmunoglobulinemia D and periodic fever syndrome (HIDS), tumor necrosis factor (TNF) receptor-associated periodic syndrome (TRAPS), adult-onset multisystem inflammatory disease (AMI), and familial mediterranean fever (FMF). Still's disease (AOSD), relapsing polychondritis, Schnitzler syndrome, Sweet syndrome, Behcet's disease, antisynthetase syndrome, interleukin-1 receptor antagonist deficiency (DIRA), tumors, type 2 diabetes, obesity, gout, atherosclerosis, rheumatoid arthritis, multiple sclerosis, amyotrophic lateral sclerosis, systemic lupus erythematosus, inflammatory bowel disease, hepatitis, alcoholic liver disease, kidney disease, silicosis, silicosis, asbestosis, asthma, acute respiratory distress syndrome, depression, Parkinson's disease, Alzheimer's disease, aging, contact hypersensitivity, blindness caused by degeneration of visual pigment cells, sepsis.

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