Tyk2 selective inhibitors and uses thereof

Abstract

The present application discloses a TYK2 selective inhibitor and its use, specifically relates to a compound of formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereoisomer or pharmaceutically acceptable salt thereof, and its use for preparing a drug for treating a disease mediated by TYK2.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of chemical medicine, and particularly relates to a TYK2 selective inhibitor and use thereof. BACKGROUND

[0002] Autoimmune diseases are a family of at least 80 diseases, such as rheumatoid arthritis, systemic lupus erythematosus and inflammatory bowel disease, and are a group of diseases in which immune cells are activated and autoantibodies are excessively produced to attack the body's own organs, tissues and cells. Autoimmune diseases affect 5-10% of people worldwide (Shoenfeld Y, Tincani A, Gershwin ME (2012) Sex gender and autoimmunity. J Autoimmun 38:J71-J73). As chronic and debilitating diseases, autoimmune diseases have high medical costs, and the quality of life of patients is reduced, which has become a huge burden to patients, their families and society. Although the pathogenesis of these diseases is not completely clear, studies have shown that various factors such as genetics, environment and immune response play an important role in the occurrence and development of the disease.

[0003] Kinases play a very important role in regulating immune cell function (Deng, Bellanti, Zheng. Essential Kinases and Transcriptional Regulators and Their Roles in Autoimmunity [J]. Biomolecules, 2019, 9(4)). Janus kinase (JAK) family members include JAK1, JAK2, JAK3, and TYK2, the non-receptor tyrosine kinases of the JAK family play an important role in mediating a variety of cytokines leading to inflammation (O'Shea J J, Schwartz D M, Villarino AV, et al. The JAK-STAT Pathway: Impact on Human Disease and Therapeutic Intervention [J]. Annual Review of Medicine, 2015, 66(1): 311-328). Genome-wide association studies show that other variants of tyrosine kinase 2 (TYK2) are associated with autoimmune diseases such as Crohn's disease, psoriasis, systemic lupus erythematosus, and rheumatoid arthritis, which further indicates the importance of TYK2 in autoimmunity (Ellinghaus D, Ellinghaus E, Nair RP, Stuart PE, Esko T, Metspalu A, et al. Combined analysis of genome-wide association studies for Crohn disease and psoriasis identifies seven shared susceptibility loci. Am J Hum Genet. (2012) 90:636-47. 10.1016 / j.ajhg.2012.02.020; Graham D S C, Akil M, Vyse TJ. Association of polymorphisms across the tyrosine kinase gene, TYK2 in UK SLE families [J]. Rheumatology, 2007(6):927-930; Eyre S, et al. High-density genetic mapping identifies new susceptibility loci for rheumatoid arthritis. Nat Genet. 2012;44:1336-40.).TYK2 is critical downstream in the regulatory signaling cascade of IL-12, IL-23 and type I interferon receptors (Karaghiosoff M, Steinborn R, Kovarik P, et al. Central role for type I interferons and Tyk2 in lipopolysaccharide-induced endotoxin shock. [J]. Nature Immunology, 2003, 4(5): 471.). IL-12 and IL-23 can activate antigen presenting cells and are able to promote differentiation and proliferation of Th1 and Th17. Human genomics studies found that IL-12R and IL-23B (encoding p40 subunit) polymorphisms have a strong correlation with inflammatory bowel disease (Stahl EA, Raychaudhuri S, Remmers EF, Xie G, Eyre S, Thomson BP, et al. Genome-wide association study meta-analysis identifies seven new rheumatoid arthritis risk loci. Nat Genet. 2010; 42(6): 508-514. R. H. Duerr, K. D; Taylor, S. R. Brant, et al. A Genome-Wide Association Study Identifies IL23R as an Inflammatory Bowel Disease Gene [J]. Science, 2006.). Type I interferons have multiple effects on the innate and adaptive immune system, including activation of cellular and humoral immunity and enhancement of the expression and release of autoantigens (Hall J C, Rosen A. Type I interferons: crucial participants in disease amplification in autoimmunity [J]. Nature Reviews Rheumatology, 2010, 6(1): 40.).Elevated serum IFN levels have been observed in patients with systemic lupus erythematosus (SLE) and the levels correlate with disease activity and severity (Bengtsson A, Sturfelt G, Truedsson L, et al. Activation of type I interferon system in systemic lupus erythematosus correlates with disease activity but not with antiretroviral antibodies [J]. Lupus, 2000, 9(9):664.). In summary, drugs that inhibit the action of IL-12, IL-23 and type I interferons have therapeutic benefits in human autoimmune diseases.

[0004] Studies have shown that selective inhibition of TYK2 activity can be a new method of balancing the relationship between treatment and safety for treating various autoimmune diseases (Leitner, Nicole, R, et al.. Tyrosine kinase 2-Surveillant of tumours and bona fide oncogene [J]. Cytokine, 2017.). TYK2 and other members of the JAK family are characterized by having dual kinase domains, respectively, tyrosine kinase domain (JH1) and pseudokinase domain (JH2). In the JAK family, the JH1 region has high sequence homology, which poses a challenge for the design of TYK2 selective inhibitors. JH2 plays an important role in the regulation of JAKs function (Lead Optimization of a 4-Aminopyridine Benzamide Scaffold To Identify Potent, Selective, and Orally Bioavailable TYK2 Inhibitors [J]. Journal of Medicinal Chemistry, 2013, 56(11):4521.), and mutations in the JH2 portion of JAKs have been shown to be associated with hematological and immunological diseases. Therefore, TYK2 JH2 selective inhibitors can more specifically inhibit TYK2 activity.

[0005] In summary, the synthesis of new TYK2 JH2 selective inhibitors can benefit patients with autoimmune diseases by regulating IL-12, IL-23 and type I interferons in vivo. SUMMARY

[0006] The application discloses a kind of compounds as TYK2 selective inhibitor and its purposes in preparing the drug for preventing or treating TYK2 mediated related diseases.

[0007] In one aspect, the application discloses a compound shown in general formula (I):

[0008]

[0009] Or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt thereof,

[0010] Wherein, A1, A2, A4, A5, A6, A7, A8 are selected from C or N, A3 is selected from C, N or And when A2 is N, at least one of A4, A5, A6, A7, A8 is N;

[0011] Saturated or unsaturated ring;

[0012] R1 is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O)2R a Or 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms can be optionally substituted by 1-3 R a ;

[0013] R2 is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a Rb , -NR a R b , -SR a , -S(O)R a , -S(O)2R a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms optionally substituted with 1-3 R a ;

[0014] R4is selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O)2R a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms optionally substituted with 1-3 R a ;

[0015] each R a , R b is independently selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, C3-C6heterocycloalkyl, halogen, hydroxy, cyano, nitro, -C(O)NR c R d , -C(O)R c , -(CH2) n C(O)OR c , -OR c , -(CH2) n OR c , -OC(O)R c , -OC(O)OR c , -OC(O)NR c R d , -NR c R d , -SR c , -S(O)R cor -S(O)2R c , said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl is optionally substituted with 1-3 R c ;

[0016] each R c , R d is independently selected from hydrogen, halogen, carbonyl, -C(O)CH3, hydroxyl, cyano, nitro, C1-C6alkyl, haloC1-C6alkyl, C3-C6cycloalkyl, or haloC3-C6cycloalkyl; and

[0017] each n is independently selected from 0, 1, 2, or 3.

[0018] In some embodiments, the compound is represented by Formula (Ia):

[0019]

[0020] or a tautomer, mesomer, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt thereof,

[0021] wherein A1, A2, A4, A5, A6, A7, A8are selected from C or N, A3is selected from C, N or and when A2is N, at least one of A4, A5, A6, A7, A8is N;

[0022] is a saturated or unsaturated ring;

[0023] R1is selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O)2R a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted with 1-3 R a ;

[0024] R2is selected from the group consisting of hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O)2R a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms being optionally substituted with 1-3 R a ;

[0025] R3is selected from the group consisting of hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O)2R a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms being optionally substituted with 1-3 R a ;

[0026] R4is selected from the group consisting of hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NRa R b , -NR a R b , -SR a , -S(O)R a , -S(O)2R a , or a 3-10 membered saturated or non-saturated ring containing 0-3 heteroatoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or non-saturated ring containing 0-3 heteroatoms optionally substituted with 1-3 R a ;

[0027] each R a , R b is independently selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, C3-C6heterocycloalkyl, halogen, hydroxyl, cyano, nitro, -C(O)NR c R d , -C(O)R c , -(CH2) n C(O)OR c , -OR c , -(CH2) n OR c , -OC(O)R c , -OC(O)OR c , -OC(O)NR c R d , -NR c R d , -SR c , -S(O)R c , or -S(O)2R c , said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl optionally substituted with 1-3 R c ;

[0028] each R c , R d is independently selected from hydrogen, halogen, carbonyl, -C(O)CH3, hydroxyl, cyano, nitro, C1-C6alkyl, haloC1-C6alkyl, C3-C6cycloalkyl, or haloC3-C6cycloalkyl; and

[0029] each n is independently selected from 0, 1, 2, or 3.

[0030] In some embodiments, the compound of Formula (I) is represented by Formula (Ib):

[0031]

[0032] or a tautomer, mesomer, racemate, enantiomer, diastereomer thereof, or a pharmaceutically acceptable salt thereof, wherein A1, A3is selected from C or N;

[0033] R1is selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O)2R a , or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms optionally substituted with 1-3 R a ;

[0034] R2is selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O)2R a , or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms optionally substituted with 1-3 R a ;

[0035] R3is selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, halogenated C1-C6alkyl, halogenated C2-C6alkenyl, halogenated C2-C6alkynyl, halogen, cyano, nitro, -C(O)NR a R b-C(O)R a -C(O)OR a -OR a -(CH2) n OR a -NR a R b -S(O)2R a ;

[0036] R4' is selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, haloC1-C6alkyl, haloC2-C6alkenyl, haloC2-C6alkynyl, C3-C6cycloalkyl, haloC3-C6cycloalkyl, halogen, cyano, nitro, or -NR a R b ;

[0037] each R a , R b is independently selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, C3-C6heterocycloalkyl, halogen, hydroxy, cyano, nitro, -C(O)NR c R d -C(O)R c -(CH2) n C(O)OR c -OR c -(CH2) n OR c -OC(O)R c -OC(O)OR c -OC(O)NR c R d -NR c R d -SR c -S(O)R c -S(O)2R c , said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl optionally substituted with 1-3 R c ;

[0038] each R c , R d is independently selected from hydrogen, halogen, carbonyl, -C(O)CH3, hydroxy, cyano, nitro, C1-C6alkyl, haloC1-C6alkyl, C3-C6cycloalkyl, or haloC3-C6cycloalkyl; and

[0039] each n is independently selected from 0, 1, 2, or 3.

[0040] In other embodiments, the compound of general formula (I) is represented by general formula (Ic):

[0041]

[0042] or a tautomer, meso, racemic, enantiomer, diastereomer thereof, or a pharmaceutically acceptable salt thereof, wherein A3 is selected from C or N;

[0043] R1is selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O)2R a , or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms optionally substituted with 1-3 R a ;

[0044] R2is selected from hydrogen, C1-C6alkyl, haloC1-C6alkyl, halogen, cyano, -C(O)OR a , -(CH2) n OR a , or -NH2;

[0045] R3is selected from hydrogen, C1-C6alkyl, or -OR a ;

[0046] R4' is selected from hydrogen, C1-C6alkyl, haloC1-C6alkyl, C3-C6cycloalkyl, halogen, or -NH2;

[0047] each R a , R b is independently selected from hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, C3-C6heterocycloalkyl, halogen, hydroxyl, cyano, nitro, -C(O)NR c R d , -C(O)R c , -(CH2) n C(O)OR c , -OR c , -(CH2)n OR c , -OC(O)R c , -OC(O)OR c , -OC(O)NR c R d , -NR c R d , -SR c , -S(O)R c or -S(O)2R c , said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl optionally substituted with 1-3 R c ;

[0048] each R c , R d is independently selected from hydrogen, halogen, carbonyl, -C(O)CH3, hydroxyl, cyano, nitro, C1-C6alkyl, haloC1-C6alkyl, C3-C6cycloalkyl or haloC3-C6cycloalkyl; and

[0049] each n is independently selected from 0, 1, 2 or 3.

[0050] In some embodiments, R1is selected from hydrogen, -CH3, -CN, -F,

[0051] In some embodiments, R2is selected from hydrogen, -CH3,

[0052] In some embodiments, R3is In some embodiments, R4is hydrogen, -C(O)CH3, or -C(O)NH2. In some embodiments, the compound of Formula (I) is selected from:

[0053]

[0054]

[0055]

[0056]

[0057]

[0058] In another aspect, the present application also provides use of the aforementioned compound, isomer thereof or pharmaceutically acceptable salt thereof for preparing a medicament for treating a disease mediated by TYK2.

[0059] In some embodiments, the TYK2-mediated disease is an autoimmune disease, an inflammatory disease, a proliferative disease, an endocrine disease, a nervous system disease, or a disease associated with transplantation.

[0060] In other embodiments, the disease is an autoimmune disease.

[0061] In other embodiments, the autoimmune disease is selected from type 1 diabetes mellitus, ankylosing spondylitis, systemic lupus erythematosus, multiple sclerosis, systemic sclerosis, psoriasis, Crohn's disease, ulcerative colitis, or inflammatory bowel disease.

[0062] In other embodiments, the disease is an inflammatory disease.

[0063] In other embodiments, the inflammatory disease is selected from rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, psoriasis, Crohn's disease, ulcerative colitis, and inflammatory bowel disease.

[0064] In another aspect, the present application also provides a pharmaceutical composition comprising a therapeutically effective amount of the aforementioned compound, isomer thereof, or pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient.

[0065] The terms:

[0066] Unless otherwise indicated, the terms used in the specification and claims have the following meanings.

[0067] The term "isomer" includes enantiomeric, diastereomeric, and geometric (or conformational) isomers of the given structure. For example, the present application includes R and S configurations at each asymmetric center, Z and E double bond isomers, Z and E conformational isomers, single stereochemical isomers, and mixtures of enantiomeric, diastereomeric, and geometric (or conformational) isomers.

[0068] The term "pharmaceutically acceptable salt" refers to, for example, acid and / or base addition salts. Suitable acid addition salts are formed from acids which form nontoxic salts, for example hydrochloride salts. Suitable base salts are formed from bases which form nontoxic salts, for example calcium and sodium salts. Also formed are half salts of acids and bases, for example, hemisulfate and hemicalcium salts.

[0069] The term "therapeutically effective amount" refers to an amount of a compound of the present application that (i) treats a particular disease, condition, or disorder; (ii) reduces, alleviates or eliminates one or more symptoms of a particular disease, condition, or disorder; or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder described herein.

[0070] The term "pharmaceutically acceptable carrier or excipient" refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated.

[0071] The term "alkyl" refers to saturated aliphatic hydrocarbon groups, which are straight-chain or branched-chain groups, preferably alkyl groups containing 1 to 20 carbon atoms, more preferably alkyl groups containing 1 to 12 carbon atoms, and even more preferably alkyl groups containing 1 to 6 carbon atoms. Non-limiting examples of lower alkyl groups containing 1 to 6 carbon atoms include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like.

[0072] The term "alkenyl" refers to aliphatic hydrocarbons having at least one carbon-carbon double bond, including straight-chain and branched-chain. In some embodiments, alkenyl groups have 2 to 20 carbon atoms, 2 to 10 carbon atoms, 2 to 6 carbon atoms, 3 to 6 carbon atoms, or 2 to 4 carbon atoms. For example, the term "C 2-6 The term "alkenyl" refers to aliphatic hydrocarbons having at least one carbon-carbon double bond, including straight-chain and branched-chain. In some embodiments, alkenyl groups have 2 to 20 carbon atoms, 2 to 10 carbon atoms, 2 to 6 carbon atoms, 3 to 6 carbon atoms, or 2 to 4 carbon atoms. For example, the term "C

[0073] The term "alkynyl" refers to aliphatic hydrocarbons having at least one carbon-carbon triple bond, including straight-chain and branched-chain. In some embodiments, alkynyl groups have 2 to 20, 2 to 10, 2 to 6, or 3 to 6 carbon atoms. For example, "C 2-6 The term "alkynyl" refers to aliphatic hydrocarbons having at least one carbon-carbon triple bond, including straight-chain and branched-chain. In some embodiments, alkynyl groups have 2 to 20, 2 to 10, 2 to 6, or 3 to 6 carbon atoms. For example, "C

[0074] The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy groups include methoxy, ethoxy, propyloxy, butyloxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy.

[0075] The term "cycloalkyl" refers to saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituents, the cycloalkyl ring comprising 3 to 20 carbon atoms, preferably comprising 3 to 12 carbon atoms, more preferably comprising 3 to 6 carbon atoms (e.g., 3, 4, 5, or 6 carbon atoms), most preferably comprising 5 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spirocyclic, fused, and bridged cycloalkyl groups.

[0076] The term "spirocycloalkyl" refers to a polycyclic group of 5 to 20 members that shares one carbon atom (termed a spiro atom) between single rings, which can contain one or more double bonds, but no ring has a fully conjugated pi-electron system. Preferably 6 to 14 members, more preferably 7 to 10 members (e.g., 7, 8, 9, or 10 members). Spirocycloalkyl groups are classified as mono-, bi-, or polycyclo, preferably mono- and bi-, according to the number of spiro atoms shared between rings. More preferably 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospriocycloalkyl groups.

[0077] The term "fused cycloalkyl" refers to a fully carbon polycyclic group of 5 to 20 members, each ring in the system sharing an adjacent pair of carbon atoms with other rings in the system, in which one or more rings can contain one or more double bonds, but no ring has a fully conjugated pi-electron system. Preferably 6 to 14 members, more preferably 7 to 10 members. Fused cycloalkyl groups are classified as bi-, tri-, tetra-, or polycyclic, preferably bi- or tri-, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicycloalkyl groups, according to the number of constituent rings.

[0078] The term "bridged cycloalkyl" refers to a fully carbon polycyclic group of 5 to 20 members, in which any two rings share two non-adjacent carbon atoms, which can contain one or more double bonds, but no ring has a fully conjugated pi-electron system. Preferably 6 to 14 members, more preferably 7 to 10 members. Bridged cycloalkyl groups are classified as bi-, tri-, tetra-, or polycyclic, preferably bi-, tri-, or tetra-, more preferably bi- or tri-, according to the number of constituent rings.

[0079] The term "heterocycloalkyl" refers to saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituents containing from 3 to 20 ring atoms, one or more of which are heteroatoms selected from nitrogen, oxygen, or S(O)m, where m is an integer from 0 to 2, but excluding cyclic moieties of -O-O-, -O-S-, or -S-S-, the remainder of the ring atoms being carbon. Preferably, there are from 3 to 12 ring atoms, of which from 1 to 4 are heteroatoms; most preferably, there are from 3 to 8 ring atoms, of which from 1 to 3 are heteroatoms; most preferably, there are from 5 to 6 ring atoms, of which from 1 to 2 or 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocycloalkyl groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like, preferably tetrahydropyranyl, piperidinyl, pyrrolidinyl. Polycyclic heterocycloalkyl groups include spiroheterocycloalkyl, fused heterocycloalkyl, and bridged heterocycloalkyl groups.

[0080] The term "spiroheterocycloalkyl" refers to a polycyclic heterocyclic group of 5 to 20 members sharing one atom between rings (referred to as a spiro atom), one or more of which are heteroatoms selected from nitrogen, oxygen, or S(O)m, where m is an integer from 0 to 2, the remainder of the ring atoms being carbon. It can contain one or more double bonds, but no ring has a fully conjugated pi-electron system. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. It can be referred to as a single spiroheterocycloalkyl, a double spiroheterocycloalkyl, or a multiple spiroheterocycloalkyl, preferably a single spiroheterocycloalkyl and a double spiroheterocycloalkyl. More preferably, it is a 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered single spiroheterocycloalkyl.

[0081] The term "fused heterocycloalkyl" refers to a polycyclic heterocyclic group of 5 to 20 members, each ring in the system sharing an adjacent pair of atoms with other rings in the system, one or more of which can contain one or more double bonds, but no ring has a fully conjugated pi-electron system, one or more of which are heteroatoms selected from nitrogen, oxygen, or S(O)m, where m is an integer from 0 to 2, the remainder of the ring atoms being carbon. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. It can be referred to as a bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocycloalkyl, preferably a bicyclic or tricyclic, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocycloalkyl.

[0082] The term "bridged heterocyclyl" refers to a 5- to 14-membered, polycyclic heterocyclic radical, sharing two non-adjacent ring atoms, which can contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen, or S(O)m (where m is an integer from 0 to 2), and the remaining ring atoms are carbon. Preferably, 6- to 14-membered, more preferably 7- to 10-membered. Depending on the number of rings, it can be referred to as bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclyl, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic.

[0083] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (that is, rings which share adjacent pairs of carbon atoms) groups having a completely conjugated pi-electron system, preferably 6- to 10-membered, such as phenyl and naphthyl.

[0084] The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5- to 10-membered, containing 1 to 3 heteroatoms; more preferably 5- or 6-membered, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furanyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazolyl, pyrazinyl and pyridazinyl.

[0085] The term "hydroxyalkyl" refers to an alkyl group, as defined above, substituted with a hydroxyl group.

[0086] The term "haloalkyl" refers to an alkyl group, as defined above, substituted with one or more halogens.

[0087] The term "haloalkoxy" refers to an alkoxy group, as defined above, substituted with one or more halogens.

[0088] The term "deuteroalkyl" refers to an alkyl group, as defined above, substituted with one or more deuterium atoms.

[0089] The term "deuteroalkoxy" refers to an alkoxy group, as defined above, substituted with one or more deuterium atoms.

[0090] The term "cycloalkylalkyl" refers to an alkyl group, as defined above, substituted with one or more cycloalkyl groups, as defined above.

[0091] The term "cycloalkyloxy" refers to -O-cycloalkyl, wherein cycloalkyl is as defined above.

[0092] The term "heterocyclylalkyl" refers to an alkyl group, as defined above, substituted with one or more heterocyclyl groups, as defined above.

[0093] The term "arylalkyl" refers to an alkyl group, as defined above, substituted with one or more aryl groups, as defined above.

[0094] The term "hydroxy" refers to an -OH group.

[0095] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0096] The term "amino" refers to -NH2.

[0097] The term "cyano" refers to -CN.

[0098] The term "nitro" refers to -NO2.

[0099] The term "carboxy" refers to -C(O)OH. DETAILED DESCRIPTION

[0100] Intermediate preparation

[0101] Preparation of Intermediate 1.1: (R)-2-bromo-6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridine

[0102]

[0103] Step one: Preparation of 3-(6-bromo-4-methylpyridin-2-yl)tetrahydrofuran-3-ol. 2,6- dibromo-4-methylpyridine (45.0 g, 179 mmol, 1.00 eq.) was dissolved in DCM (500 mL), n-BuLi (197 mmol, 78.9 mL, 1.10 eq.) was added dropwise at -78 °C, stirred for 1 h, dihydrofuran-3(2H)-one (16.98 g, 197.28 mmol, 1.1 eq.) was added, reacted at -78 °C for 0.5 h, and then naturally warmed to room temperature for 1 h. 350 mL saturated ammonium chloride solution was added, after separation, 50 mL saturated sodium chloride was used for washing, and concentrated. The crude product was purified by column chromatography (PE / EtOAc = 5:1-1:1) to give the title compound (31 g, yield 67%). MS (m / z) = 258.01 [M+H] + .

[0104] Step two: Preparation of (R)-2-bromo-6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridine. 3-(6-bromo-4-methylpyridin-2-yl)tetrahydrofuran-3-ol (29.0 g, 112 mmol, 1.00 eq.) was dissolved in DMF (180 mL), NaH (3.00 g, 125 mmol, 1.1 eq.) was added, CH3I (17.5 g, 124 mmol, 1.1 eq.) was added, and the reaction was stirred at 25 °C for 1.5 h. The reaction was poured into 400 mL of ice water, and EA (100 mL*3) was added. The crude product was purified by column chromatography (PE / EtOAc = 5:1-3:1) and then chiral resolution to give the title compound (RT = 2.26 min, 12 g, 40% yield). MS (m / z) = 272.02 [M+H] + .

[0105] Chiral resolution conditions:

[0106] Instrument: Waters 200, preparative SFC (QC-R-LC-07); column: ChiralPak IC, 250 x 30 mm I.D., 5 μm;

[0107] Mobile phase: A: CO2 and B: Ethanol (0.1% NH3H2O); Gradient: B 15%; Flow rate: 70 mL / min; Back pressure: 100 bar; Column temperature: 35 °C; Wavelength: 254 nm; Cycle time: ~1.5 min; Interval time: 0.5 min; Sample preparation: Compound dissolved in 200 mL ethanol.

[0108] Intermediates 1.2-1.6 were prepared according to the procedure described for Intermediate 1.1.

[0109]

[0110]

[0111] Preparation of Intermediate 2.1: 6-chloro-3-vinyl-1H-pyrrolo[3,2-c]pyridine

[0112]

[0113] Step one: Preparation of 3-bromo-6-chloro-1H-pyrrolo[3,2-c]pyridine. 6-chloro-1H- pyrrolo[3,2-c]pyridine (20 g, 131 mmol, 1.0 eq.) was dissolved in DMF (120 mL), NBS (23 g, 131 mmol, 1 eq.) was added, and the reaction was stirred at 25 °C for 12 h. The reaction was poured into 600 mL of water, and a large amount of solid was precipitated, which was filtered and dried. The title compound was obtained (30.3 g, 99%). MS (m / z) = 230.92 [M+H]+ .

[0114] Step two: Preparation of 6-chloro-3-vinyl-lH-pyrrolo[3,2-c]pyridine. 3-bromo-6-chloro-lH-pyrrolo[3,2-c]pyridine (1.00 g, 4.32 mmol, 1 eq.), 4,4,5,5-tetramethyl-2-vinyl-l,3,2-dioxaborolane (665 mg, 4.32 mmol, 1 eq.), Pd(dppf)Cl2(3.16 g, 4.32 mmol, 1 eq.), K2CO3(597 mg, 4.32 mmol, 1 eq.) dissolved in water (2 mL) and dioxane (10 mL), 100 °C for 3 h under nitrogen protection. The reaction was rotary evaporated, the crude product was purified by column chromatography (PE / EtOAc = 10:1-1:1) to give the title compound (0.7 g, yield 90%). MS (m / z) = 179.03 [M+H] + .

[0115] Intermediates 2.2-2.9 were prepared according to the procedure described for Intermediate 2.1.

[0116]

[0117]

[0118] Preparation of Intermediate 3: 6-chloro-3-cyclopropyl-lH-pyrrolo[3,2-c]pyridine

[0119]

[0120] Step one: Preparation of 6-chloro-3-iodo-lH-pyrrolo[3,2-c]pyridine. 6-chloro-lH-pyrrolo[3,2-c]pyridine (2.00 g, 13.1 mmol, 1 eq.) was dissolved in DMF (12 mL), NIS (2.95 g, 13.1 mmol, 1 eq.) was added portionwise at 0 °C, and the reaction was stirred at 25 °C for 12 h. The reaction was poured into 60 mL of water, and a large amount of solid was precipitated, which was filtered and dried to give the title compound (3.50 g, yield 96%). MS (m / z) = 278.91 [M+H] + .

[0121] Step two: Preparation of 6-chloro-3-iodo-l-((2-(trimethylsilyl)ethoxy)methyl)- lH-pyrrolo[3,2-c]pyridine. 6-chloro-3-iodo-lH-pyrrolo[3,2-c]pyridine (3.50 g, 12.6 mmol, 1 eq.) was added to DMF (20 mL) at 0 °C, NaH (331 mg, 13.8 mmol, 1.1 eq.) was added and stirred for 15 min, then SemCl (2.10 g, 12.6 mmol, 1 eq.) was added and reacted at 25 °C for 2 h. The reaction was poured into 100 mL water, extracted with 50 mL EA, the organic phase was dried and filtered. The crude product was purified by column chromatography (PE / EtOAc = 10:1-1:1) to give the title compound (5 g, yield 97%). MS (m / z) = 408.99 [M+H] + .

[0122] Step three: Preparation of 6-chloro-3-cyclopropyl-l-((2-(trimethylsilyl)ethoxy)methyl)- lH-pyrrolo[3,2-c]pyridine. Cyclopropylboronic acid (84 mg, 978 pmol, 2.0 eq.), 6-chloro-3-iodo-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2- c]pyridine (0.20 g, 489 pmol, 1 eq.), tricyclohexylphosphine (27.4 mg, 97.9 pmol, 0.2 eq.), Pd(OAc)2(11 mg, 48.9 pmol, 0.1 eq.) were dissolved in a mixture of toluene (5 mL) and water (0.2 mL), and reacted at 100 °C for 12 h. The crude product was purified by column chromatography (PE / EtOAc = 10:1-1:1) to give the title compound (0.15 g, yield 94%). MS (m / z) = 323.13 [M+H] + .

[0123] Step four: Preparation of 6-chloro-3-cyclopropyl-lH-pyrrolo[3,2-c]pyridine. 6-chloro-3- cyclopropyl-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-c]pyridine (0.15 g, 464 pmol, 1 eq.) was dissolved in 10 mL of a solution of tetrabutylammonium fluoride in THF (1 M) and refluxed for 10 h. Water was added and extracted with EA, the crude product was purified by column chromatography (PE / EtOAc = 10:1-1:1) to give the title compound (0.10 g, yield 94%). MS (m / z) = 193.05 [M+H] + .

[0124] Preparation of intermediate 4: 6-chloro-3-(l-methyl-l,2,3,6-tetrahydropyridin-4-yl)- lH-pyrrolo[3,2-c]pyridine

[0125]

[0126] 6-chloro-lH-pyrrolo[3,2-c]pyridine (370 mg, 3.28 mmol, 1 eq.) was dissolved in MeOH (5 mL), KOH (184 mg, 3.28 mmol, 1 eq.) was added, 1-methylpiperidin-4-one (370 mg, 3.28 mmol, 1 eq.) was added, and the reaction was stirred at 70 °C for 12 h. The reaction was concentrated, 50 mL water and 100 mL EA were added, the solution was partitioned, and the organic phase was concentrated to give the title compound (0.70 g, yield 86%). MS (m / z) = 248.09 [M+H] + .

[0127] Preparation of Intermediate 5: 3-(6-chloro-lH-pyrrolo[3,2-c]pyridin-3-yl)-l- methylazetidin-3-ol

[0128]

[0129] 6-chloro-lH-pyrrolo[3,2-c]pyridine (279 mg, 2.29 mmol, 0.7 eq.) was dissolved in MeOH (5 mL), KOH (184 mg, 3.28 mmol, 1 eq.) was added, 1-methylazetidin-3-one hydrochloride (370 mg, 3.28 mmol, 1 eq.) was added, and the reaction was stirred at 70 °C for 12 h. The reaction was concentrated, 50 mL water and 100 mL EA were added, the solution was partitioned, and the organic phase was concentrated to give the title compound (0.20 g, yield 26%). MS (m / z) = 238.07 [M+H] + .

[0130] Synthesis of Intermediate 6: 3-(6-chloro-lH-pyrrolo[3,2-c]pyridin-3-yl)bicyclo[l.l.l] pentane-l-carbonitrile

[0131]

[0132] Step 1: Preparation of 3-cyano-N-methoxy-N-methylbicyclo[1.1.1]pentane-1-carboxamide. 3-cyanobicyclo[1.1.1]pentane-1-carboxylic acid (2.00 g, 14.6 mmol, 1 eq.), N,O-dimethylhydroxylamine hydrochloride (1.42 g, 14.6 mmol, 1 eq.), HATU (6.65 g, 17.5 mmol, 1.20 eq.), and DIEA (3.77 g, 29.2 mmol, 2.00 eq.) were dissolved in DCM (20 mL) and reacted at 20 °C for 12 h. 10 mL of citric acid was added to the reaction solution, and after separation, the mixture was washed once with 10 mL of saturated sodium carbonate, then once with 10 mL of saturated sodium chloride, and concentrated to obtain the title compound (2 g, yield 76%).

[0133] Step 2: Preparation of 3-(4,6-dichloronicotinyl)bicyclo[1.1.1]pentane-1-onitrile. 5-Bromo-2,4-dichloropyridine (2.10 g, 9.25 mmol, 1 eq.) was dissolved in THF (20 mL), and isopropyl magnesium chloride (10.2 mmol, 5.09 mL, 1.1 eq.) was added dropwise at -78 °C. The reaction was allowed to proceed for 2 h, followed by the addition of 3-cyano-N-methoxy-N-methylbicyclo[1.1.1]pentane-1-carboxamide (2.00 g, 11.10 mmol, 1.20 eq.) at -78 °C. The mixture was allowed to rise naturally to room temperature and reacted for 10 h. 20 mL of saturated ammonium chloride solution was added to the reaction mixture, and the mixture was concentrated after separation. The crude product was purified by column chromatography (PE / EtOAc = 10:1–1:1) to give the title compound (1.5 g, 60% yield). MS(m / z) = 267.00 [M+H] + .

[0134] Step 3: Preparation of 3-(6-chloro-1H-pyrazolo[4,3-c]pyridin-3-yl)bicyclo[1.1.1]pentane-1-onitrile. 3-(4,6-dichloronicotinyl)bicyclo[1.1.1]pentane-1-onitrile (1.50 g, 5.62 mmol, 1 eq.) was dissolved in THF (20 mL), and hydrazine hydrate (562 mg, 8.42 mmol, 1.5 eq.) was added at 25 °C. After reacting for 12 h, the reaction solution was poured into 10 mL of water, and 10 mL of EA was added for extraction. The crude product was purified by column chromatography (PE / EtOAc = 10:1-2:1) to give the title compound (1.00 g, yield 72.8%). MS (m / z) = 245.05 [M+H] + .

[0135] Synthesis of Intermediate 7: 3-(6-chloro-1H-pyrrolo[3,2-c]pyridin-3-yl)azacyclobutane-1-carboxylic acid tert-butyl ester

[0136]

[0137] Step one: 3-bromo-6-chloro-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2- c]pyridine was prepared according to the method of Intermediate 2, step two. MS (m / z) = 361.01 [M+H] + .

[0138] Step two: Preparation of 3-(6-chloro-l-((2-(trimethylsilyl)ethoxy)methyl)tert-butyl-l- pyrrolo[3,2-c]pyridin-3-yl)-3-hydroxyazetidine-l-carboxylate. 3-bromo-6-chloro-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-c]pyridine (5.00 g, 13.8 mmol, 1 eq.) was dissolved in THF (30 mL), n-BuLi (14.5 mmol, 3.20 mL, 1.05 eq.) was added dropwise at -78 °C, and the mixture was stirred at -78 °C for 1 h. Then 3-oxoazetidine-l- carboxylate tert-butyl ester (2.37 g, 13.8 mmol, 1 eq.) was added dropwise at -78 °C, and the mixture was stirred at -78 °C for 1 h. The reaction was quenched with saturated ammonium chloride, and the crude product was purified by column chromatography (PE / EtOAc = 10: 1-2: 1) to give the title compound (3.50 g, yield 55.8%). MS (m / z) = 454.19 [M+H] + .

[0139] Step three: Preparation of 3-(6-chloro-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2- c]pyridin-3-yl)-3-(((methylthio)carbonothioyl)oxy)azetidine-l-carboxylate. 3-(6-chloro-l- ((2-(trimethylsilyl)ethoxy)methyl)tert-butyl-l-pyrrolo[3,2-c]pyridin-3-yl)-3-hydroxyazetidine-l- carboxylate (3.50 g, 7.71 mmol, 1 eq.) was dissolved in THF (50 mL), and NaHMDS (2.83 g, 15.4 mmol, 2.0 eq.) was added dropwise at -78 °C. CS2 (1.17 g, 15.4 mmol, 2.0 eq.) was added at -78 °C, and the mixture was stirred at -78 °C for 1 h. CH3I (2.19 g, 15.4 mmol, 2.0 eq.) was added, and the mixture was stirred at 20 °C for 2 h. The reaction was quenched with 50 mL of saturated ammonium chloride solution, and the mixture was extracted with 50 mL of EA. The crude product was purified by column chromatography (PE / EtOAc = 10: 1-3: 1) to give the title compound (3.00 g, yield 71.51%). MS (m / z) = 544.14 [M+H] + .

[0140] Step four: Preparation of tert-butyl 3-(6-chloro-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-c]pyridin-3-yl)azetidine-l- carboxylate. tert-Butyl 3-(6-chloro-l-((2-(trimethylsilyl)ethoxy)methyl)-lH- pyrrolo[3,2-c]pyridin-3-yl)-3-(((methylthio)carbonothioyl)oxy)azetidine-l- carboxylate (3.00 g, 5.51 mmol, 1 eq.) was dissolved in toluene (60 mL), AIBN (90.5 mg, 551 μmol, 0.1 eq.), Bu3SnH (2.41 g, 8.27 mmol, 1.5 eq.) were added, and the reaction was stirred at 85 °C for 3 h under nitrogen. The crude product was purified by column chromatography (PE / EtOAc = 10:1-3:1) to give a colorless liquid (2.0 g, 82% yield). MS (m / z) = 438.19 [M+H] + .

[0141] Step five: Preparation of tert-butyl 3-(6-chloro-lH-pyrrolo[3,2-c]pyridin-3- yl)azetidine-l-carboxylate. tert-Butyl 3-(6-chloro-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrrolo[3,2-c]pyridin-3-yl)azetidine-l- carboxylate (2.0 g, 1.0 eq.) was dissolved in a solution of tetrabutylammonium fluoride in THF (20 mL) and stirred at 80 °C for 5 h. The reaction was diluted with 50 mL of water and extracted with 50 mL of EA. The organic layer was separated and concentrated. The crude product was purified by column chromatography (PE / EtOAc = 50:1-1:1) to give the title compound (1.00 g, 71.16% yield). MS (m / z) = 308.11 [M+H] + .

[0142] Synthesis of Intermediate 8: tert-Butyl 6-(6-chloro-lH-pyrrolo[3,2-c]pyridin-3- yl)-2-azaspiro[3.3]heptane-2-carboxylate

[0143]

[0144] Intermediate 8 was prepared according to the procedure described for Intermediate 7. MS (m / z) = 348.14 [M+H] + .

[0145] Example 1: N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3- vinyl-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0146]

[0147] Step one: Preparation of 3-bromo-6-chloro-lH-pyrrolo[3,2-c]pyridine. 6-chloro-lH-pyrrolo[3,2-c]pyridine (20 g, 131 mmol, 1.0 eq.) was dissolved in DMF (120 mL), NBS (23 g, 131 mmol, 1 eq.) was added, and the reaction was stirred at 25 °C for 12 h. The reaction was poured into 600 mL of water, and a large amount of solid was precipitated, which was filtered and dried. The title compound was obtained (30.3 g, 99%). MS (m / z) = 230.92 [M+H] + .

[0148] Step two: Preparation of 6-chloro-3-vinyl-lH-pyrrolo[3,2-c]pyridine. 3-bromo-6-chloro-lH-pyrrolo[3,2-c]pyridine (1.00 g, 4.32 mmol, 1 eq.), 4,4,5,5-tetramethyl-2-vinyl-l,3,2-dioxaborolane (665 mg, 4.32 mmol, 1 eq.), Pd(dppf)Cl2(3.16 g, 4.32 mmol, 1 eq.), K2CO3(597 mg, 4.32 mmol, 1 eq.) were dissolved in water (2 mL) and dioxane (10 mL), and the reaction was stirred at 100 °C for 3 h under nitrogen protection. The reaction was rotary evaporated, and the crude product was purified by column chromatography (PE / EtOAc = 10:1-1:1) to obtain the title compound (0.7 g, yield 90%). MS (m / z) = 179.03 [M+H] + .

[0149] Step three: Preparation of 6-chloro-l-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-3-vinyl-lH-pyrrolo[3,2-c]pyridine. 2-bromo-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine (152 mg, 559 μmol, 1 eq.), 6-chloro-3-vinyl-lH-pyrrolo[3,2-c]pyridine (0.1 g, 559 μmol, 1 eq.), K2CO3(232 mg, 1.68 mmol, 3.0 eq.), CuI (5.33 mg, 27.99 μmol, 0.05 eq.), N,N-dimethylethylenediamine (4.94 mg, 56 μmol, 0.1 eq.) were dissolved in 1,4-dioxane (30 mL), and the reaction was refluxed for 12 h under nitrogen protection. The reaction was rotary evaporated, and the crude product was purified by column chromatography (PE / EtOAc = 5:1-1:1) to obtain the title compound (0.2 g, yield 97%). MS (m / z) = 370.12 [M+H] + .

[0150] Step four: Preparation of N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-3-vinyl-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide. 6-chloro-l-(6-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-vinyl-lH-pyrrolo[3,2-c]pyridine (0.20 g, 540 pmol, 1 eq.), acetamide (47.9 mg, 811 pmol, 1.5 eq.), Cs2CO3(528 mg, 1.62 mmol, 3.0 eq.), Pd2(dba)3(49.5 mg, 54.1 pmol, 0.1 eq.), X-phos (52 mg, 108 pmol, 0.2 eq.) were dissolved in 1,4-dioxane (30 mL) and refluxed under nitrogen for 12 h. The reaction was concentrated and the crude product was purified by column chromatography (PE / EtOAc = 2: 1-0: 1) to give the title compound (0.2 g, yield 94%). MS (m / z) = 393.18 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.45 (s, 1H), 9.08 (s, 1H), 8.94 (s, 1H), 8.19 (s, 1H), 7.56 (s, 1H), 7.34 (s, 1H), 6.92 (dd, 1H), 5.92 (d, 1H), 5.33 (d, 1H), 4.20 (d, 1H), 4.08 - 3.82 (m, 3H), 3.15 (s, 3H), 2.74 - 2.62 (m, 1H), 2.47 (s, 4H), 2.11 (s, 3H).

[0151] The compounds of Examples 2-20 were prepared according to the procedure of Example 1.

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160] Example 21 : N-(3-ethyl-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H- pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0161]

[0162] N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-vinyl-1H-pyrrolo[3,2- c]pyridin-6-yl)acetamide (0.05 g, 127 μmol, 1 eq.) was dissolved in MeOH (10 mL), replaced with hydrogen three times, and reacted at 20 °C for 12 h. The reaction solution was filtered, and the crude product was purified by column chromatography (PE / EtOAc = 2:1-0:1) to obtain the title compound (0.05 g, yield 99%). MS (m / z) = 395.20 [M+H] + . 1 HNMR (400 MHz, DMSO) δ 10.38 (s, 1H), 9.06 (s, 1H), 8.63 (s, 1H), 7.80 (s, 1H), 7.53 (s, 1H), 7.27 (s, 1H), 4.20 (d, 1H), 4.05 - 3.88 (m, 3H), 3.14 (s, 3H), 2.80 (q, 2H), 2.68 (dt, 1H), 2.46 (s, 4H), 2.10 (s, 3H), 1.35 (t, 3H).

[0163] The compounds of Examples 22-27 were prepared according to the method of Example 21.

[0164]

[0165]

[0166]

[0167] Example 28a or Example 28b: N-(3-((1s,3s)-3-cyanocyclobutyl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide or N-(3-((1r,3r)-3-cyanocyclobutyl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0168]

[0169] Step one: Preparation of tert-butyl 3-bromo-6-chloro-lH-pyrrolo[3,2-c]pyridine-l- carboxylate. 3-bromo-6-chloro-lH-pyrrolo[3,2-c]pyridine (2.00 g, 8.64 mmol, 1 eq.), (Boc)20 (1.89 g, 8.64 mmol, 1.0 eq.), DMAP (105 mg, 864 μmol, 0.1 eq.), TEA (1.31 g, 12.9 mmol, 1.5 eq.) were dissolved in THF (20 mL) and reacted at 25 °C for 1 h. The crude product was purified by column chromatography (PE / EtOAc = 10:1-3:1) to give the title compound (2.80 g, 97.7%). MS (m / z) = 330.98 [M+H] + .

[0170] Step two: Preparation of tert-butyl 6-chloro-3-(3-cyano-l-hydroxycyclobutyl)-lH- pyrrolo[3,2-c]pyridine-l-carboxylate. tert-butyl 3-bromo-6-chloro-lH-pyrrolo[3,2- c]pyridine-l-carboxylate (2.80 g, 8.44 mmol, 1 eq.) was dissolved in THF (30 mL), n-BuLi (8.87 mmol, 3.20 mL, 1.05 eq.) was added dropwise at -78 °C, and the mixture was incubated at -78 °C for 1 h. 3-oxocyclobutanenitrile (883 mg, 9.29 mmol, 1.1 eq.) was added dropwise at -78 °C, and the mixture was reacted at -78 °C for 1 h. The reaction solution was quenched with saturated ammonium chloride, dried after being separated into layers, filtered, and concentrated. The crude product was purified by column chromatography (PE / EtOAc = 5:1-1:1) to give the title compound (2.00 g, 68.10%). MS (m / z) = 348.10 [M+H] + .

[0171] Step three: Preparation of 3-(6-chloro-lH-pyrrolo[3,2-c]pyridin-3-yl)cyclobutan-l- enitrile. tert-butyl 6-chloro-3-(3-cyano-l-hydroxycyclobutyl)-lH-pyrrolo[3,2- c]pyridine-l-carboxylate (2.00 g, 5.75 mmol, 1 eq.), Et3SiH (6.69 g, 57.5 mmol, 10.0 eq.), TFA (6.56 g, 57.5 mmol, 10.0 eq.) were dissolved in DCM (30 mL) and reacted at 25 °C for 1 h. The reaction solution was rotary evaporated, 10 mL of saturated sodium carbonate solution and 10 mL of ethyl acetate were added, the organic phase was dried after being separated into layers, filtered, and concentrated to give the title compound (1.00 g, 75.06%). MS (m / z) = 232.06 [M+H] + .

[0172] Step four: Preparation of 3-(6-chloro-l-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-3-yl)cyclobutan-l -one. 2-Bromo-6-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridine (1.17 g, 4.32 mmol, 1 eq.), 3-(6-chloro- lH-pyrrolo[3,2-c]pyridin-3-yl)cyclobutan-l-one (1.00 g, 4.32 mmol, 1 eq.), K2CO3 (1.79 g, 12.9 mmol, 3.0 eq.), Cul (1.10 mg, 216 μmol, 0.05 eq.), N,N- dimethylethylenediamine (38.1 mg, 431 μmol, 0.1 eq.) were dissolved in 1,4-dioxane (30 mL) and refluxed under nitrogen for 12 h. The reaction was concentrated and the crude product was purified by column chromatography (PE / EtOAc = 5:1-1:1) to give the title compound (1 g, 55% yield). MS (m / z) = 423.15 [M+H] + .

[0173] Step five: Preparation of N-(3-(3-cyanocyclobutyl)-l-(6-(3-methoxytetrahydrofuran-3- yl)-4-methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide. 3-(6-chloro-l-(6-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-3-yl)cyclobutan- 1-one (1.00 g, 2.36 mmol, 1 eq.), acetamide (209 mg, 3.55 mmol, 1.5 eq.), Cs2CO3 (2.31 g, 7.09 mmol, 3.0 eq.), Pd2(dba)3 (216 mg, 236 μmol, 0.1 eq.), X-phos (225 mg, 473 μmol, 0.2 eq.) were dissolved in 1,4-dioxane (50 mL) and refluxed under nitrogen for 12 h. The reaction was concentrated and the crude product was purified by column chromatography (EtOAc) to give the title compound (0.2 g, 19% yield). MS (m / z) = 446.21 [M+H] + .

[0174] Step six: The compound from Step five was purified by preparative TLC (eluent: EtOAc, Rf f 1 = 0.43, R f 2 = 0.36) to give the compounds of Example 28a and Example 28b. 1H NMR (400 MHz, DMSO) δ 10.38 (s, 1H), 9.07 (s, 1H), 8.58 (s, 1H), 8.02 (s, 1H), 7.61 (s, 1H), 7.30 (s, 1H), 4.21 (d, J = 9.7 Hz, 1H), 4.09 - 3.94 (m, 2H), 3.92 (d, 1H), 3.81 (dt, 1H), 3.51 - 3.37 (m, 1H), 3.15 (s, 3H), 2.87 (ddd, 2H), 2.68 (dt, 2H), 2.57 (dd, 2H), 2.47 (s, 3H), 2.10 (s, 3H). 1 H NMR (400 MHz, DMSO) δ 10.38 (s, 1H), 9.07 (s, 1H), 8.58 (s, 1H), 8.02 (s, 1H), 7.61 (s, 1H), 7.30 (s, 1H), 4.21 (d, J = 9.7 Hz, 1H), 4.09 - 3.94 (m, 2H), 3.92 (d, 1H), 3.81 (dt, 1H), 3.51 - 3.37 (m, 1H), 3.15 (s, 3H), 2.87 (ddd, 2H), 2.68 (dt, 2H), 2.57 (dd, 2H), 2.47 (s, 3H), 2.10 (s, 3H).

[0175] Example 29a and Example 29b were prepared in analogy to the procedures described for Example 28a and Example 28b (eluent: EtOAc, R f 1 = 0.43, R f 2 = 0.36).

[0176]

[0177] Example 30: (R)-2-(4-(6-acetylamino-1-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-3-yl)cyclohexyl)acetic acid

[0178]

[0179] Step one: Preparation of (R)-2-(4-(6-acetylamino-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-3-yl)cyclohexyl)acetic acid. (R)-2-(4-(6-acetylamino-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-3-yl)cyclohex-3-en-1-yl)acetic acid ethyl ester was prepared from Intermediate 2.5 and Intermediate 1.1 by a similar method as in Example 1, and then by a similar method as in Example 22 to give the title compound. MS (m / z) = 535.28 [M+H] + .

[0180] Step two: Preparation of (R)-2-(4-(6-acetylamino-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-3-yl)cyclohexyl)acetic acid. (R)-2-(4-(6-acetylamino-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-3-yl)cyclohexyl)acetic acid ethyl ester (0.30 g, 561 μmol, 1 eq.) was dissolved in H2O (3 mL) and THF (9 mL) with LiOH·H2O (47.1 mg, 1.12 mmol, 2.0 eq.), and stirred at 20 °C for 12 h. The THF was removed from the reaction solution, and the remaining liquid was adjusted to pH 5 with 1 N HCl, and a large amount of solid was precipitated, which was filtered and dried. The title compound (0.2 g, yield 70%) was obtained. MS (m / z) = 507.25 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.38 (d, 1H), 9.06 (s, 1H), 8.66 (d, 1H), 7.75 (d, 1H), 7.58 (d, 1H), 7.27 (s, 1H), 4.21 (d, 1H), 4.05 - 3.77 (m, 3H), 3.16 (s, 3H), 2.94 (s, 1H), 2.79 (s, 1H), 2.72 - 2.59 (m, 2H), 2.46 (s, 3H), 2.21 - 2.02 (m, 5H), 2.00 - 1.92 (m, 2H), 1.88 (d, 1H), 1.80 (d, 3H), 1.68 - 1.47 (m, 2H).

[0181] Example 31 was prepared by a similar method as in Example 30.

[0182]

[0183]

[0184] Example 32: (R)-N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3- (piperidin-4-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0185]

[0186] Step one: Preparation of (R)-4-(6-acetamido-l-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-3-yl)piperidine-l-carboxylic acid tert-butyl ester. (R)-4-(6-acetamido-l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-lH- pyrrolo[3,2-c]pyridin-3-yl)-3,6-dihydropyridine-l(2H)-carboxylic acid tert-butyl ester was prepared from Intermediate 2.7 and Intermediate 1.1 by a similar method as Example 1, and then by a similar method as Example 22 to give the title compound. MS (m / z) = 550.30 [M+H] + .

[0187] Step two: Preparation of (R)-N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)- 3-(piperidin-4-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide. (R)-4-(6-acetamido-l-(6-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-3-yl)piperidine- 1-carboxylic acid tert-butyl ester (500 mg, 909 pmol, 1 eq.) was dissolved in DCM (5 mL) and TFA (1 mL) and reacted at 20 °C for 1 h. The reaction was adjusted to pH 10 with saturated sodium carbonate and 20 mL of EA was added to extract, the solution was separated and rotary evaporated to dryness. The title compound (0.2 g, yield 49%) was obtained. MS (m / z) = 450.24 [M+H] + . 1 H NMR (400 MHz, DMSO) d 10.40 (s, 1H), 9.05 (s, 1H), 8.72 (s, 1H), 7.77 (s, 1H), 7.56 (s, 1H), 7.29 (s, 1H), 4.20 (d, 1H), 4.08 - 3.85 (m, 4H), 3.19 - 3.07 (m, 5H), 3.00 (t, 1H), 2.77 (t, 2H), 2.69 - 2.61 (m, 2H), 2.46 (s, 3H), 2.10 (s, 3H), 2.01 (d, 2H), 1.70 (dd, 2H).

[0188] Example 33 was prepared in analogy to the procedure described in Example 32.

[0189]

[0190] Example 34a or 34b: N-(3-((1s,4S)-4-aminocyclohexyl)-1-(6-((R)-3- methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6- yl)acetamide or N-(3-((1r,4R)-4-aminocyclohexyl)-1-(6-((R)-3-methoxytetrahydrofuran- 3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0191]

[0192] Example 34a and 34b were prepared from the compound of Example 33 via chiral resolution (RT1= 3.38 min, RT2= 4.19 min).

[0193] Chiral resolution conditions:

[0194] Instrument: Waters 200, preparative SFC (QC-R-LC-07); column: ChiralPak IC, 250 x 30 mm I.D., 5 μm; mobile phase: A is CO2 and B is ethanol (0.1% NH3H2O); gradient: B 30%; flow rate: 70 mL / min; back pressure: 100 bar; column temperature: 35 °C; wavelength: 210 nm; cycle time: ~ 4 min; interval time: 0.5 min; sample preparation: compound dissolved in 200 mL ethanol.

[0195] 1 H NMR (400 MHz, DMSO) δ 10.37 (s, 1H), 9.05 (s, 1H), 8.70 (s, 1H), 7.78 (s, 1H), 7.56 (s, 1H), 7.29 (s, 1H), 4.20 (d, 1H), 4.10 - 3.82 (m, 3H), 3.78 - 3.54 (m, 4H), 3.16 (s, 3H), 2.97 (s, 1H), 2.73 - 2.60 (m, 1H), 2.46 (s, 3H), 2.10 (s, 3H), 1.99 (t, 2H), 1.83 - 1.60 (m, 4H), 1.40 (d, 2H). MS (m / z) = 464.26 [M+H] + .

[0196] 1H NMR (400 MHz, DMSO) δ 10.37 (s, 1H), 9.04 (s, 1H), 8.70 (s, 1H), 7.75 (s, 1H), 7.54 (s, 1H), 7.28 (s, 1H), 4.20 (d, 1H), 4.07 - 3.87 (m, 3H), 3.71 (s, 1H), 3.64 (d, 1H), 3.14 (s, 3H), 2.67 (dd, 4H), 2.45 (s, 3H), 2.10 (s, 3H), 2.06 (s, 2H), 1.97 - 1.84 (m, 2H), 1.60 (dd, 2H), 1.33 (d, 2H). MS (m / z) = 464.26 [M+H] + .

[0197] Example 35: (R)-N-(3-(1-ethylpiperidin-4-yl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0198]

[0199] (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(piperidin-4-yl)-1H- pyrrolo[3,2-c]pyridin-6-yl)acetamide (100 mg, 222 μmol, 1 eq.) was dissolved in 37% acetaldehyde aqueous solution (2.73 mL) and DCM (2.27 mL), stirred for 1 h, then STAB (377 mg, 445 μmol, 2.0 eq.) was added, and the reaction was carried out at 20 °C for 14 h. The reaction solution was adjusted to pH 10 with saturated sodium carbonate solution, 20 mL EA was added for extraction, the organic phase was dried, filtered, and rotary evaporated to obtain the title compound (0.1 g, yield 94%). MS (m / z) = 478.27 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.41 (s, 1H), 9.05 (s, 1H), 8.84 (s, 1H), 7.83 (s, 1H), 7.57 (s, 1H), 7.31 (s, 1H), 4.20 (d, 1H), 3.98 (ddd, 3H), 3.59 (s, 2H), 3.15 (s, 7H), 2.66 (dt, 2H), 2.47 (s, 4H), 2.25 (d, 2H), 2.11 (s, 5H), 1.36 - 1.21 (m, 3H).

[0200] Example 36a: (R)-N-(3-(azetidin-3-yl)-l-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide and Example 36b: (R)-N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(l- methylazetidin-3-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0201]

[0202] The title compound was obtained according to the procedure described in Reference Example 1 and Example 35.

[0203] 36a: MS (m / z) = 422 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.39 (s, 1H), 9.06 (s, 1H), 8.76 (s, 1H), 7.93 (s, 1H), 7.56 (s, 1H), 7.29 (s, 1H), 4.20 (d, 1H), 4.13 (t, 1H), 4.02 (m, 2H), 3.91 (m, 3H), 3.80 (m, 2H), 3.33 (s, 1H), 3.14 (s, 3H), 2.68 (m, 1H), 2.49 (m, 4H), 2.11 (s, 3H).

[0204] 36b: MS (m / z) = 436.23 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.42 (s, 1H), 9.07 (s, 1H), 8.68 (s, 1H), 8.17 (s, 1H), 7.56 (s, 1H), 7.34 (s, 1H), 4.39 (s, 2H), 4.31 - 4.19 (m, 2H), 4.12 (s, 2H), 3.98 (ddd, 3H), 3.15 (s, 3H), 2.83 (s, 3H), 2.68 (dt, 1H), 2.49 (s, 4H), 2.11 (s, 3H).

[0205] Example 37a: (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(2- azaspiro[3.3]heptan-6-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide and Example 37b: (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(2-methyl-2- azaspiro[3.3]heptyl-6-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0206]

[0207] The title compound was obtained in reference example 1 and example 35.

[0208] 37a: MS (m / z) = 462 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.40 (s, 1H), 9.04 (s, 1H), 9.00 (s, 1H), 8.56 (s, 1H), 7.86 (s, 1H), 7.56 (s, 1H), 7.29 (s, 1H), 4.19 (d, 1H), 4.13 (m, 1H), 3.98 (m, 5H), 3.60 (m, 1H), 3.33 (s, 1H), 3.14 (s, 3H), 2.76 (m, 2H), 2.68 (m, 1H), 2.49 (m, 6H), 2.10 (s, 3H).

[0209] 37b: MS (m / z) = 476.26 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.35 (s, 1H), 9.04 (s, 1H), 8.55 (s, 1H), 7.85 (s, 1H), 7.55 (s, 1H), 7.29 (s, 1H), 4.31 - 4.19 (m, 8H), 3.14-3.20 (m, 4H), 2.83 (s, 3H), 2.52-2.68 (m, 5H), 2.49 (s, 4H), 2.11 (s, 3H).

[0210] Example 38: (R)-N-(3-(1-acetylpiperidin-4-yl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0211]

[0212] (R)-N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(piperidin-4- yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide (0.30 g, 739 μmol, 1 eq.) was dissolved in DCM (10 mL), TEA (149 mg, 1.48 mmol, 2.0 eq.) and acetyl chloride (69.7 mg, 888 μmol, 1.2 eq.) were added, after reaction at 20 °C for 1 h, the reaction solution was poured into 10 mL water, liquid-liquid separation and rotary evaporation to obtain the title compound (0.3 g, yield 82%). MS (m / z) = 492.25 [M+H] + . 1 H NMR (400 MHz, CDC13) δ 8.94 (s, 1H), 8.59 (s, 1H), 8.21 (s, 1H), 7.43 (s, 1H), 7.29 (s, 1H), 7.21 (s, 1H), 4.11-4.30 (m, 4H), 3.05-3.25 (m, 4H), 2.75-2.85 (m, 6H), 1.80-2.50 (m, 13H).

[0213] Examples 39-40 were prepared by analogy with the method of Example 38.

[0214]

[0215]

[0216] Example 41a or Example 41b: N-(7-((ls,3S)-3-cyanocyclobutyl)-5-(6-((R)-3- methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-7H-pyrrolo[2,3-c]pyridazin-3- yl)acetamide or N-(7-((lr,3R)-3-cyanocyclobutyl)-5-(6-((R)-3-methoxytetrahydrofuran- 3-yl)-4-methylpyridin-2-yl)-7H-pyrrolo[2,3-c]pyridazin-3-yl)acetamide

[0217]

[0218] Step one: Preparation of 5-bromo-3-chloro-7H-pyrrolo[2,3-c]pyridazine. 3-chloro-7H- pyrrolo[2,3-c]pyridazine (2.00 g, 13.0 mmol, 1 eq.) was dissolved in DMF (12 mL), NBS (2.32 g, 13.0 mmol, 1 eq.) was added, and the reaction was carried out at 20 °C for 2 h. The reaction solution was poured into 60 mL water, a large amount of solid was precipitated, which was filtered and the filter cake was dried to obtain the title compound (3 g, yield 99%). MS (m / z) = 231.92 [M+H] + .

[0219] Step two: Preparation of 3-cyanocyclobutyl methanesulfonate. 3- hydroxycyclobutyl nitrile (1.50 g, 15.4 mmol, 1 eq.), MsCl (2.12 g, 18.5 mmol, 1.2 eq.), TEA (2.34 g, 23.1 mmol, 1.5 eq.) were dissolved in DCM (20 mL) and reacted at 20 °C for 2 h. The reaction was poured into 10 mL water, partitioned and concentrated to give the title compound (2.7 g, yield 99%).

[0220] Step three: Preparation of 3-(5-bromo-3-chloro-7H-pyrrolo[2,3-c]pyridazin-7- yl)cyclobutan-1 -carbonitrile. 5-bromo-3-chloro-7H-pyrrolo[2,3-c]pyridazine (1.19 g, 5.14 mmol, 1 eq.), 3-cyanocyclobutyl methanesulfonate (2.70 g, 15.4 mmol, 3.0 eq.), Cs2CO3(3.35 g, 10.2 mmol, 2.0 eq.) were dissolved in DMF (10 mL) and reacted at 80 °C for 12 h. The reaction was poured into 50 mL water, a large amount of solid was precipitated, filtered, and the filter cake was dried to give the title compound (1.2 g, yield 75%). MS (m / z) = 310.96 [M+H] + .

[0221] Step four: Preparation of (R)-3-(3-chloro-5-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-7H-pyrrolo[2,3-c]pyridazin-7-yl)cyclobutan-1 -carbonitrile. 3-(5-bromo-3-chloro-7H-pyrrolo[2,3-c]pyridazin-7-yl)cyclobutan-1 -carbonitrile (500 mg, 1.60 mmol, 1 eq.), coupling pinacol borate (407 mg, 1.60 mmol, 1.0 eq.), Pd(dppf)Cl2DCM (131 mg, 160 μmol, 0.1 eq.), potassium acetate (315 mg, 3.21 mmol, 2.0 eq) were dissolved in 10 mL dioxane, protected by nitrogen, and reacted at 110 °C for 2 h. The reaction was cooled to room temperature, potassium phosphate (681 mg, 3.21 mmol, 2.0 eq) was added, 10 mL dioxane, 2-bromo-6-(3(R)-methoxy-tetrahydrofuran-3-yl)-4-methyl-pyridine (437 mg, 1.60 mmol, 1 eq) and water (2 mL) were added, and the reaction was continued at 110 °C for 1 h. The reaction was directly prepared sand, and the crude product was purified by column chromatography (PE / EtOAc = 2:1-1 :1) to give the title compound (0.5 g, yield 74%) as a white solid. MS (m / z) = 424.15 [M+H] + .

[0222] Step five: Preparation of (R)-N-(7-(3-cyanocyclobutyl)-5-(6-(3-methoxytetrahydrofuran- 3-yl)-4-methylpyridin-2-yl)-7H-pyrrolo[2,3-c]pyridazin-3-yl)acetamide. (S)-3-(3-chloro-5-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-7H- pyrrolo[2,3-c]pyridazin-7-yl)cyclobutan-1 -carbonitrile (500 mg, 1.18 mmol, 1 eq.), cesium carbonate (1.15 g, 3.54 mmol, 3.0 eq.), acetamide (104 mg, 1.77 mmol, 1.5 eq.), Pd2(dba)3 (108 mg, 118 μmol, 0.1 eq.), X-phos (23.0 mg, 236 μmol, 0.2 eq.) were dissolved in 1,4-dioxane (5 mL) and stirred at 110 °C for 15 h under nitrogen atmosphere. The crude product was purified by column chromatography (EtOAc) to give the title compound (0.1 g, 19% yield). MS (m / z) = 447.21 [M+H] + .

[0223] Step six: The compound from step five was purified by preparative TLC (eluent: EtOAc, R f 1 = 0.45, R f 2 = 0.42) to give the compounds of Example 41a and Example 41b. 1 1H NMR (400 MHz, DMSO) δ 10.93 (s, 1H), 9.34 (s, 1H), 8.95 (s, 1H), 7.72 (s, 1H), 7.20 (s, 1H), 5.86 - 5.63 (m, 1H), 4.27 - 4.15 (m, 1H), 4.08 - 3.97 (m, 2H), 3.89 (d, 1H), 3.68 - 3.53 (m, 1H), 3.21 - 3.06 (m, 5H), 3.01 - 2.88 (m, 2H), 2.72 (dt, 1H), 2.42 (s, 4H), 2.18 (s, 3H). 1 1H NMR (400 MHz, DMSO) δ 10.93 (s, 1H), 9.34 (s, 1H), 8.95 (s, 1H), 7.72 (s, 1H), 7.20 (s, 1H), 5.86 - 5.63 (m, 1H), 4.27 - 4.15 (m, 1H), 4.08 - 3.97 (m, 2H), 3.89 (d, 1H), 3.68 - 3.53 (m, 1H), 3.21 - 3.06 (m, 5H), 3.01 - 2.88 (m, 2H), 2.72 (dt, 1H), 2.42 (s, 4H), 2.18 (s, 3H).

[0224] Example 42: N-(3-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)imidazo[l,5- a]pyrazin-6-yl)acetamide

[0225]

[0226] Step one: Preparation of 6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine carboxylic acid. 2-bromo-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine (1.00 g, 3.67 mmol, 1 eq.) was dissolved in THF (10 mL), n-BuLi (3.67 mmol, 1.5 mL, 1.0 eq.) was added dropwise at -78 °C, after 0.5 h, CO2was bubbled in, and the reaction was allowed to warm to room temperature for 1 h. The reaction solution was added to 10 mL of water, and the solution was concentrated to give the title compound (0.8 g, yield 91%). MS (m / z) = 238.10 [M+H] + .

[0227] Step two: Preparation of tert-butyl ((5-chloropyrazin-2-yl)methyl)carbamate. 5-chloropyrazine-2- carboxaldehyde (1.00 g, 7.02 mmol, 1 eq.), Et3SiH (8.16 g, 70.1 mmol, 10.0 eq.), NH2Boc (1.64 g, 14.03 mmol, 2.0 eq.), TFA (2.40 g, 21.0 mmol, 3.0 eq.) were dissolved in ACN (10 mL), and the reaction was allowed to proceed at 25 °C for 24 h. 10 mL of saturated sodium carbonate was added, and the ACN was removed by rotary evaporation. The solution was extracted with 10 mL of EA, and the solution was concentrated to give the title compound (1.2 g, yield 70%). MS (m / z) = 244.08 [M+H] + .

[0228] Step three: Preparation of (5-chloropyrazin-2-yl)methanamine. tert-butyl ((5-chloropyrazin-2- yl)methyl)carbamate (1.20 g, 4.92 mmol, 1 eq.) was dissolved in a solution of HCl in EA (10 mL), and the reaction was allowed to proceed at 25 °C for 16 h. The solution was concentrated to give the title compound (0.8 g, yield 100%). MS (m / z) = 144.03 [M+H] + .

[0229] Step four: Preparation of N-((5-chloropyrazin-2-yl)methyl)-6-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridinamide. (5-Chloropyrazin-2- yl)methanamine (181 mg, 1.26 mmol, 3.0 eq.), 6-(3-methoxytetrahydrofuran-3- yl)-4-methylpyridinecarboxylic acid (100 mg, 0.42 mmol, 1.0 eq.), HATU (480 mg, 1.26 mmol, 3.0 eq.), DIEA (544 mg, 4.2 mmol, 10.0 eq.) were dissolved in 10 mL DCM, and reacted at room temperature for 15 h. 10 mL water was added, and after separation, the title compound (0.1 g, yield 65%) was purified by column chromatography (PE / EtOAc = 2: 1-1: 1). MS (m / z) = 363.11 [M+H] + .

[0230] Step five: Preparation of 6-chloro-3-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)imidazo[l,5-a]pyrazine. N-((5-chloropyrazin-2-yl)methyl)-6-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridinamide (0.10 g, 275 μmol, 1 eq.) was dissolved in ACN (5 mL), and POCl3(211 mg, 1.38 mmol, 5.0 eq.) was added, and reacted at 90 °C for 3 h. After quenching with 10 mL water and extraction with dichloromethane, the title compound (0.1 g, yield 95%) was obtained. MS (m / z) = 345.10 [M+H] + .

[0231] Step six: Preparation of N-(3-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)imidazo[l,5-a]pyrazin-6-yl)acetamide. 6-chloro-3-(6-(3-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)imidazo[l,5-a]pyrazine (0.10 g, 290 μmol, 1 eq.), cesium carbonate (283 mg, 870 μmol, 3.0 eq.), acetamide (25.7 mg, 435 μmol, 1.5 eq.), Pd2(dba)3(26.5 mg, 29.0 μmol, 0.1 eq.), X-phos (5.70 mg, 58.0 μmol, 0.2 eq.) were dissolved in 1,4-dioxane (5 mL), and reacted at 100 °C for 12 h under nitrogen protection. The reaction solution was concentrated, and the title compound (0.08 g, yield 75%) was purified by column chromatography (PE / EtOAc = 2: 1-0: 1). MS (m / z) = 368.16 [M+H] + . 1HNMR (400 MHz, DMSO) δ 10.50 (s, 1H), 10.27 (s, 1H), 9.10 (d, 1H), 8.10 (s, 1H), 8.03 (d, 1H), 7.40 (s, 1H), 4.31 (d, 1H), 4.10 - 3.97 (m, 2H), 3.95 (d, 1H), 3.14 (s, 3H), 2.76 (dt, 1H), 2.59 - 2.53 (m, 1H), 2.47 (s, 3H), 2.14 (s, 3H).

[0232] Example 43: N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3- methyl-2-oxo-2,3-dihydro-lH-imidazo[4,5-c]pyridin-6-yl)acetamide

[0233]

[0234] Step one: Preparation of 6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2- amine. 2-bromo-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine (0.50 g, 1.84 mmol, 1 eq.), dimethylethylenediamine (16.2 mg, 183 pmol, 0.1 eq.), Cu20 (13.1 mg, 91.8 pmol, 0.05 eq.), potassium carbonate (508 mg, 3.67 mmol, 2.0 eq.) were dissolved in ethylene glycol (5 mL) and ammonia water (4.6 mL), and reacted at 80 °C for 12 h under nitrogen protection. 20 mL EA and 20 mL water were added to the reaction solution, which was separated, concentrated and purified by column chromatography (PE / EtOAc = 2: 1-0: 1) to obtain the title compound (0.3 g, yield 78%). MS (m / z) = 209.12 [M+H] + .

[0235] Step two: Preparation of N-(2-chloro-5-nitropyridin-4-yl)-6-(3-methoxytetrahydrofuran- 3-yl)-4-methylpyridin-2-amine. 2,4-dichloro-5-nitropyridine (278 mg, 1.44 mmol, 1 eq.), 6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-amine (0.30 g, 1.44 mmol, 1 eq.), DIPEA (186 mg, 1.44 mmol, 1 eq.) were dissolved in ethanol (10 mL), and reacted at 78 °C for 12 h. Purification by column chromatography (PE / EtOAc = 5: 1-1: 1) to obtain the title compound (0.5 g, yield 95%). MS (m / z) = 365.09 [M+H] + .

[0236] Step three: 6-chloro-N4 Preparation of (6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)pyridin-3,4-diamine. N-(2-chloro-5-nitropyridin-4-yl)-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-amine (0.50 g, 1.37 mmol, 1 eq.) and Fe (383 mg, 6.85 mmol, 5 eq.) were dissolved in saturated ammonium chloride solution (10 mL) and ethanol (10 mL) and reacted at 70 °C for 3 h. It was dried, extracted with EA, and purified by column chromatography (PE / EtOAc = 5:1-1:1) to obtain the title compound (0.4 g, yield 87%). MS (m / z) = 335.12 [M+H] + .

[0237] Step four: Preparation of 6-chloro-l-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-l,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one. 6-chloro-N 4 Preparation of (6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)pyridin-3,4-diamine. N-(2-chloro-5-nitropyridin-4-yl)-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-amine (0.50 g, 1.37 mmol, 1 eq.) and Fe (383 mg, 6.85 mmol, 5 eq.) were dissolved in saturated ammonium chloride solution (10 mL) and ethanol (10 mL) and reacted at 70 °C for 3 h. It was dried, extracted with EA, and purified by column chromatography (PE / EtOAc = 5:1-1:1) to obtain the title compound (0.4 g, yield 87%). MS (m / z) = 335.12 [M+H] + .

[0238] Step five: Preparation of 6-chloro-l-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-3-methyl-l,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one. 6-chloro-l-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-l,3-dihydro- 2H-imidazo[4,5-c]pyridin-2-one (0.30 g, 831 μmol, 1 eq.) was dissolved in DMF (2 mL), NaH (20 mg, 831 μmol, 1 eq.) was added at 20 °C, and after 15 min, iodomethane (118 mg, 831 μmol, 1 eq.) was added, and the reaction was continued for 2 h. The reaction solution was poured into 10 mL of water and extracted with 10 mL of EA, and the liquid was separated and concentrated to obtain the title compound (0.2 g, yield 64%). MS (m / z) = 375.11 [M+H] + .

[0239] Step six: Preparation of N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-3-methyl-2-oxo-2,3-dihydro-lH-imidazo[4,5-c]pyridin-6-yl)acetamide. 6-chloro-l-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-methyl- l,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (0.20 g, 534 pmol, 1 eq.), acetamide (37.8 mg, 640 pmol, 1.2 eq.), Pd2(dba)3(49 mg, 53 pmol, 0.1 eq.), X-phos (50.87 mg, 106 pmol, 0.2 eq.), cesium carbonate (522 mg, 1.60 mmol, 3 eq.) were dissolved in 1,4-dioxane (5 mL) and reacted at 100 °C for 12 h under nitrogen protection. Concentration and purification by column chromatography (PE / EtOAc = 2: 1-0: 1) gave the title compound (0.2 g, yield 94%). MS (m / z) = 398.18 [M+H] + . 1 H NMR (400 MHz, DMSO) d 10.41 (s, 1H), 8.62 (s, 1H), 8.23 (s, 1H), 7.78 (s, 1H), 7.40 (s, 1H), 4.16 (d, 1H), 3.95 (dd, 2H), 3.90 (d, 1H), 3.44 (s, 3H), 3.13 (s, 3H), 2.59 (dd, 1H), 2.45 (d, 3H), 2.44 (d, 1H), 2.07 (s, 3H).

[0240] Example 44: (R)-N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3- (oxetan-3-ylethynyl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0241]

[0242] Step one: Preparation of (R)-6-chloro-l-(6-(3-(methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridine. 6-chloro-lH-pyrrolo[3,2-c]pyridine (6.000 g, 39.324 mmol, 1.00 eq.) was dissolved in 1,4-dioxane (80 mL), (R)-2-bromo-6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridine (10.702 g, 39.324 mmol, 1.00 eq.), K2CO3(16.305 g, 0.118 mol, 3.0 eq.), Cul (0.375 mg, 1.966 mmol, 0.05 eq.), N,N-dimethylethylenediamine (0.348 g, 3.933 mmol, 0.1 eq.), refluxed under nitrogen protection for 12 h. The reaction solution was rotary evaporated, and the crude product was purified by column chromatography (PE / EtOAc = 1:1) to give the title compound (11.040 g, yield 81.65%). MS (m / z) = 344 [M+H] + .

[0243] Step two: Preparation of (R)-N-(l-(6-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide. (R)-6-chloro-l-(6-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridine (9.820 g, 28.562 mmol, 1.0 eq.) was dissolved in 1,4-dioxane (100 mL), acetamide (2.531 g, 42.843 mmol, 1.5 eq.), Pd2(dba)3(2.615 g, 2.856 mmol, 0.1 eq.), X-phos (2.723 g, 5.713 mmol, 0.2 eq.), Cs2CO3(18.612 g, 57.124 mmol, 2.0 eq.), 100 °C for 5 h under nitrogen protection. The reaction solution was cooled, poured into water (150 mL), extracted with EA (200 mL), separated, the organic phase was washed once with saturated NaCl aqueous solution, separated, the organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography (PE / EtOAc = 1:1) to give the title compound (9.500 g, yield 90.70%). MS (m / z) = 367 [M+H] + .

[0244] Step three: Preparation of (R)-N-(3-iodo-l-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide. (R)-N-(l-(6-(6-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide (9.500 g, 25.927 mmol, 1.0 eq.) was dissolved in DMF (50 mL), NIS (8.166 g, 36.298 mmol, 1.4 eq.) was added, and the reaction was stirred at 60 °C for 1 h. The reaction was cooled, poured into water (150 mL), and extracted with EA (200 mL). The organic phase was washed once with an aqueous solution of sodium sulfite, once with saturated aqueous NaCl, dried over anhydrous magnesium sulfate, filtered, and concentrated. The title compound was obtained by column chromatography (PE / EtOAc = 1:1) (6.400 g, 35.81% yield). MS (m / z) = 493 [M+H] + .

[0245] Step four: Preparation of (R)-N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-3-(oxetan-3-ylethynyl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide. (R)-N-(3-iodo-l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-lH-pyrrolo[3,2- c]pyridin-6-yl)acetamide (0.200 g, 0.408 mmol, 1.0 eq.) was dissolved in triethylamine (10 mL), 3-ethynyloxetane (0.0504 g, 0.612 mmol, 1.5 eq.), Pd(PPh3)2Cl2(0.0288 g, 0.0408 mmol, 0.1 eq.), and CuI (0.0308 g, 0.1632 mmol, 0.4 eq.) were added, and the reaction was stirred at 28 °C for 15 h under nitrogen protection. The reaction was poured into water (20 mL) and extracted with DCM (50 mL). The organic phase was washed once with saturated aqueous NH4Cl, once with saturated aqueous NaCl, dried over anhydrous magnesium sulfate, filtered, and concentrated. The title compound was obtained by column chromatography (DCM / MeOH = 30:1) (0.080 g, 43.90% yield). MS (m / z) = 447 [M+H] + . 1H NMR (400 MHz, DMSO) δ 10.54 (s, 1H), 9.09 (s, 1H), 8.68 (d, 1H), 8.40 (s, 1H), 7.63 (s, 1H), 7.37 (s, 1H), 6.68 (s, 1H), 4.85 (dd, 2H), 4.65 (dd, 2H), 4.20 (dd, 1H), 3.98 (m, 3H), 3.15 (s, 3H), 2.67 (m, 1H), 2.46 (m, 4H), 2.11 (s, 3H).

[0246] Example 45: (R)-N-(3-((3-hydroxyoxetan-3-yl)ethynyl)-l-(6-(3-methoxytetrahydrofuran- 3-yl)-4-methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0247]

[0248] The title compound was obtained following the synthetic procedure of Reference Example 44. MS (m / z) = 463 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.54 (s, 1H), 9.09 (s, 1H), 8.68 (d, 1H), 8.40 (s, 1H), 7.63 (s, 1H), 7.37 (s, 1H), 6.68 (s, 1H), 4.85 (dd, 2H), 4.65 (dd, 2H), 4.20 (dd, 1H), 3.98 (m, 3H), 3.15 (s, 3H), 2.67 (m, 1H), 2.46 (m, 4H), 2.11 (s, 3H).

[0249] Example 46: (R)-N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3- ((3-methyloxetan-3-yl)ethynyl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0250]

[0251] The title compound was obtained following the synthetic procedure of Reference Example 44. MS (m / z) = 463 [M+H] + . 1H NMR (400 MHz, DMSO) δ 10.52 (s, 1H), 9.09 (s, 1H), 8.65 (d, 1H), 8.34 (s, 1H), 7.62 (s, 1H), 7.35 (s, 1H), 4.84 (d, 2H), 4.49 (d, 2H), 4.20 (dd, 1H), 4.00 (m, 3H), 3.15 (s, 3H), 2.67 (m, 1H), 2.46 (m, 4H), 2.11 (s, 3H), 1.71 (s, 3H).

[0252] Example 47: (R)-N-{3-(1-acetyl-azetidin-3-ylethynyl)-1-[6-(3-methoxy-tetrahydrofuran-3-yl)-4-methyl-pyridin-2-yl]-1H-pyrrolo[3,2-c]pyridin-6-yl}-acetamide

[0253]

[0254] Step one: Preparation of (R)-3-((6-acetylamino-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-3-yl)ethynyl)azetidine-1-carboxylic acid tert-butyl ester. The title compound was synthesized according to the procedure of Example 44. MS (m / z) = 546 [M+H] + .

[0255] Step two: Preparation of (R)-N-(3-(azetidin-3-ylethynyl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide. (R)-3-((6-acetylamino-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-3-yl)ethynyl)azetidine-1-carboxylic acid tert-butyl ester (0.15 g, 0.275 mmol, 1.0 eq.) was dissolved in DCM (10 mL), TFA (4 mL) was added at room temperature, and stirred for 30 min. The reaction mixture was poured into 30 mL of water, and the pH was adjusted to 10 with sodium carbonate. The mixture was extracted with 100 mL of DCM, and the organic phase was washed once with saturated aqueous NH4Cl, once with saturated aqueous NaCl, dried over anhydrous magnesium sulfate, filtered, and concentrated to give the title compound (0.12 g, 95% yield). MS (m / z) = 446 [M+H] + .

[0256] Step 3: Preparation of (R)-N-(3-((l-acetylazetidin-3-yl)ethynyl)-l-(6-(3- methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-6- yl)acetamide. (R)-N-(3-(azetidin-3-ylethynyl)-l-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide (0.12 g, 0.269 mmol, 1.0 eq.) was dissolved in DCM (10 mL), acetic anhydride (0.042 g, 0.405 mmol, 1.5 eq.) was added at room temperature, and the reaction was stirred for 30 min. The system was poured into 10 mL of water, the pH was adjusted to 10 with saturated aqueous sodium carbonate solution, extracted with 50 mL of DCM, separated, the organic phase was washed once with saturated aqueous NH4C1 solution, separated, the organic phase was washed once with saturated aqueous NaCl solution, separated, the organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography (DCM / MeOH = 30: 1) to give the title compound (0.045 g, yield 34.4%). MS (m / z) = 488 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.52 (s, 1H), 9.07 (s, 1H), 8.66 (d, 1H), 8.34 (s, 1H), 7.60 (s, 1H), 7.36 (s, 1H), 4.48 (t, 1H), 4.22 (m, 3H), 4.00 (m, 4H), 3.80 (m, 1H), 3.15 (s, 3H), 2.67 (m, 1H), 2.46 (m, 4H), 2.11 (s, 3H), 1.80 (s, 3H).

[0257] Example 48: (R)-N-(3-((3-methyloxetan-3-ylethynyl)-l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0258]

[0259] The title compound was synthesized according to the procedure of Reference Example 44. MS (m / z) = 477 [M+H] + . 1H NMR (400 MHz, DMSO) δ 10.54 (s, 1H), 9.09 (s, 1H), 8.68 (s, 1H), 8.47 (s, 1H), 7.64 (s, 1H), 7.38 (s, 1H), 4.84 (d, 2H), 4.70 (d, 2H), 4.20 (m, 1H), 3.98 (m, 3H), 3.41 (s, 3H), 3.15 (s, 3H), 2.67 (m, 1H), 2.46 (m, 4H), 2.11 (s, 3H).

[0260] Example 49: (R)-N-(3-(4-hydroxybut-1-yn-1-yl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0261]

[0262] The title compound was synthesized according to the procedure of Reference Example 44. MS (m / z) = 435 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.54 (s, 1H), 9.09 (s, 1H), 8.68 (s, 1H), 8.47 (s, 1H), 7.64 (s, 1H), 7.38 (s, 1H), 4.84 (d, 2H), 4.70 (d, 2H), 4.20 (m, 1H), 3.98 (m, 3H), 3.41 (s, 3H), 3.15 (s, 3H), 2.67 (m, 1H), 2.46 (m, 4H), 2.11 (s, 3H).

[0263] Example 50: (R)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(1- methylazetidin-3-yl)-1H-pyrrolo[3,2-c]pyridin-6-amine

[0264]

[0265] (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(1- methylazetidin-3-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide (0.10 g, 0.229 mmol, 1.0 eq.) was dissolved in methanol (10 mL), 4M aqueous sodium hydroxide solution (5 mL) was added, heated to 60 °C, and reacted for 15 h. The system was rotary evaporated, and the title compound was purified by column chromatography (DCM / MeOH = 30:1). MS (m / z) = 394 [M+H] + . 1H NMR (400 MHz, DMSO) δ 8.33 (s, 1H), 7.70 (s, 1H), 7.48 (s, 1H), 7.28 (s, 1H), 7.22 (s, 1H), 5.66 (s, 2H), 4.21 (m, 2H), 4.01 (m, 2H), 3.89 (m, 4H), 3.50 (s, 2H), 3.11 (s, 3H), 2.48 (m, 4H), 2.45 (s, 3H).

[0266] Example 51 : (R)-N-(3-(1 -acetylazetidin-3-yl)-1 -(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-1 H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0267]

[0268] The title compound was synthesized by the procedure of Reference Example 47. MS (m / z) = 464 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.42 (s, 1H), 9.09 (s, 1H), 8.59 (s, 1H), 8.09 (s, 1H), 7.59 (s, 1H), 7.30 (s, 1H), 4.62 (t, 1H), 4.30 (m, 2H), 4.21 (d, 1H), 4.01 (m, 5H), 3.14 (s, 3H), 2.68 (m, 1H), 2.44 (m, 4H), 2.11 (s, 3H), 1.81 (s, 3H).

[0269] Example 52 was prepared by an analogous procedure to Example 51.

[0270]

[0271]

[0272] Example 53: (R)-N-(3-(1 -isopropylazetidin-3-yl)-1 -(6-(3-methoxytetrahydrofuran-3-yl)-4- methylpyridin-2-yl)-1 H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0273]

[0274] The title compound was synthesized by the procedure of Reference Example 35. MS (m / z) = 464 [M+H] + . 1H NMR (400 MHz, DMSO) δ 10.41 (s, 1H), 9.07 (s, 1H), 8.69 (s, 1H), 8.09 (s, 1H), 7.57 (s, 1H), 7.32 (s, 1H), 4.20 (d, 1H), 4.00 (m, 5H), 3.31 (s, 4H), 3.14 (s, 3H), 2.68 (m, 1H), 2.44 (m, 4H), 2.11 (s, 3H), 1.08 (m, 6H).

[0275] Example 54: (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(1-(2,2,2- trifluoroethyl)azetidin-3-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0276]

[0277] (R)-N-(3-(azetidin-3-yl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H- pyrrolo[3,2-c]pyridin-6-yl)acetamide (0.125 g, 0.291 mmol, 1.0 eq.) was dissolved in THF (10 mL), added 2,2,2-trifluoroethyl trifluoromethanesulfonate (0.081 g, 0.349 mmol, 1.2 eq.), triethylamine (0.0589 g, 0.582 mmol, 2.0 eq.) and reacted at room temperature for 15 h. The system was poured into 20 mL of water, extracted with 80 mL of ethyl acetate, separated, the organic phase was washed once with saturated aqueous NH4Cl solution, separated, the organic phase was washed once with saturated aqueous NaCl solution, separated, the organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography (DCM / MeOH = 30:1) to obtain the title compound (0.04 g, yield 27%). MS (m / z) = 504 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.39 (s, 1H), 9.06 (s, 1H), 8.75 (t, 1H), 7.96 (s, 1H), 7.56 (s, 1H), 7.29 (s, 1H), 4.21 (t, 1H), 4.00 (m, 5H), 3.51 (t, 2H), 3.30 (m, 3H), 3.14 (s, 3H), 2.68 (m, 1H), 2.48 (m, 4H), 2.11 (s, 3H).

[0278] Example 55 was prepared in analogy to the method described for Example 54.

[0279]

[0280]

[0281] Example 56: N-(3-(2-methyl-2-azaspiro[3.3]heptyl-6-yl)-l-(4-methylpyridin-2-yl)- lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0282]

[0283] The title compound was synthesized according to the procedure of Reference Examples 1 and 35. MS (m / z) = 376 [M+H] + . 1 HNMR (400 MHz, DMSO) δ 10.38 (s, 1H), 8.99 (d, 1H), 8.55 (d, 1H), 8.40 (d, 1H), 7.85 (d, 1H), 7.61 (s, 1H), 7.17 (d, 1H), 4.16 (s, 2H), 3.97 (s, 2H), 3.64 (t, 1H), 2.75 (m, 5H), 2.48 (m, 5H), 2.09 (s, 3H).

[0284] Example 57: (R)-l-(l-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(l- methylazetidin-3-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)urea

[0285]

[0286] (R)-l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(l-methylazetidin-3- yl)-lH-pyrrolo[3,2-c]pyridin-6-amine (0.20 g, 0.508 mmol, 1.0 eq.) was dissolved in acetic acid (10 mL) and water (30 mL), warmed to 90 °C, potassium cyanate (2.06 g, 25.41 mmol, 50 eq.) was added portionwise over an hour, the reaction was allowed to run for 20 min. The reaction was cooled to room temperature, the pH was adjusted to around 9 with potassium carbonate, the organic phase was extracted with dichloromethane, the organic phases were combined and concentrated, the title compound was purified by column chromatography (DCM / MeOH = 30: 1) (0.01 g, 4.5% yield). MS (m / z) = 437 [M+H] + . 1HNMR (400 MHz, CDC13) δ 9.71 (s, 1H), 8.50 (s, 1H), 7.85 (s, 1H), 7.73 (s, 1H), 7.31 (s, 1H), 7.25 (s, 1H), 4.30 (m, 6H), 4.05 (m, 2H), 3.25 (s, 3H), 2.86 (s, 3H), 2.81 (m, 1H), 2.51 (s, 3H), 2.48 (m, 2H), 1.60 (m, 2H).

[0287] Example 58: (R)-1-(1-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(2-methyl-2-azaspiro[3.3]heptan-6-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)urea

[0288]

[0289] The title compound was synthesized according to the procedure of Reference Examples 50 and 57. MS (m / z) = 477 [M+H] + . 1 HNMR (400 MHz, CDC13) δ 9.71 (s, 1H), 8.50 (s, 1H), 7.85 (s, 1H), 7.73 (s, 1H), 7.31 (s, 1H), 7.25 (s, 1H), 4.30 (m, 6H), 4.05 (m, 2H), 3.25 (s, 3H), 2.86 (s, 3H), 2.81 (m, 1H), 2.51 (s, 3H), 2.48 (m, 2H), 1.60 (m, 2H).

[0290] Example 59: (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(1H-pyrazol-5-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0291]

[0292] The title compound was synthesized according to the procedure of Reference Examples 1 and 44. MS (m / z) = 433 [M+H] + . 1H NMR (400 MHz, DMSO) δ 12.89 (s, 1H), 10.47 (s, 1H), 9.23 (s, 1H), 9.12 (s, 1H), 8.44 (s, 1H), 7.84 (d, 1H), 7.63 (s, 1H), 7.33 (s, 1H), 6.80 (s, 1H), 4.22 (d, 1H), 4.00 (m, 3H), 3.17 (s, 3H), 2.68 (m, 1H), 2.48 (m, 4H), 2.11 (s, 3H).

[0293] The compounds of Examples 60-62 were prepared by analogy with the method of Example 35.

[0294]

[0295]

[0296] The compounds of Examples 63-64 were prepared by the method of Example 1.

[0297]

[0298] Example 65 (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(1- methylazetidin-3-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)cyclopropanecarboxamide

[0299]

[0300] The title compound was prepared by analogy with the synthesis of Reference Examples 1 and 35. MS (m / z) = 462 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.73 (s, 1H), 9.06 (s, 1H), 8.69 (s, 1H), 8.19 (s, 1H), 7.57 (s, 1H), 7.31 (s, 1H), 4.35 (m, 2H), 4.24 (m, 1H), 4.15 (m, 1H), 4.13 (m, 1H), 3.99 (m, 2H), 3.88 (m, 2H), 3.14 (s, 3H), 2.80 (s, 3H), 2.68 (m, 1H), 2.49 (m, 4H), 2.03 (m, 1H), 1.79 (m, 4H).

[0301] Example 66 Methyl 4-(6-acetylamino-3-(2-methyl-2-azaspiro[3.3]heptan-6-yl)-1H- pyrrolo[3,2-c]pyridin-1-yl)-2,6-dimethylbenzoate

[0302]

[0303] The title compound was prepared according to the procedure described in Reference Example 56. MS (m / z) = 447 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.44 (s, 1H), 8.58 (s, 1H), 8.33 (s, 1H), 7.52 (s, 1H), 7.32 (s, 2H), 4.01 (s, 2H), 3.89 (s, 3H), 3.82 (s, 2H), 3.60 (m, 1H), 2.72 (m, 2H), 2.65 (s, 3H), 2.43 (m, 2H), 2.33 (s, 6H), 2.07 (s, 3H).

[0304] Example 67 (R)-N-(3-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)imidazo[l,5- a]pyrazin-6-yl)acetamide

[0305]

[0306] The title compound was prepared according to the procedure described in Reference Example 42. MS (m / z) = 368.42 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 10.51 (s, 1H), 10.26 (s, 1H), 9.10 (m, 1H), 8.09 (s, 1H), 8.02 (m, 1H), 7.39 (m, 1H), 3.13 (s, 3H), 2.90-2.93 (m, 2H), 2.46 (s, 3H), 1.70 (m, 2H), 1.32-1.37 (m, 2H).

[0307] The compounds of Examples 68-69 were prepared according to the procedure described in Reference Example 1.

[0308]

[0309] Example 70 (R)-N-(l-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(2-(methyl-d3)-2- azaspiro[3.3]heptan-6-yl)-lH-pyrrolo[3,2-c]pyridin-6-yl)acetamide

[0310]

[0311] (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(2- azaspiro[3.3]heptan-6-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide (0.100 g, 216.659 pmol, 1 eq) was dissolved in DMF (10 mL), NaHCO3(36.402 mg, 433.318 pmol, 2 eq.) was added, deuterated methyl iodide (21.984 mg, 151.661 pmol, 0.7 eq.), and the reaction was allowed to proceed at room temperature for 15 h. The reaction was poured into 50 mL of water and extracted with 100 mL of EtOAC. The organic phase was washed once with saturated aqueous NaCl solution, separated, dried over anhydrous magnesium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography using DCM:MeOH = 20:1 to give the title compound (0.002 g). MS (m / z) = 479.61 [M+H] + . 1 H NMR (400 MHz, MeOD) d 8.98 (s, 1H), 8.53 (s, 1H), 7.75 (m, 1H), 7.45 (s, 1H), 7.35 (m, 1H), 4.26-4.33 (m, 3H), 4.08-4.15 (m, 4H), 3.73-3.77 (m, 1H), 3.27 (s, 3H), 2.86-2.91 (m, 2H), 2.78-2.81 (m, 1H), 2.56-2.62 (m, 5H), 2.21 (s, 3H), 2.05 (s, 1H), 1.31 (s, 1H).

[0312] Examples 71 and 72 were prepared by analogy with the method of Example 1 and Example 56.

[0313]

[0314]

[0315] Examples 73a and 73b were prepared by analogy with the method of Reference Examples 28a and 28b.

[0316]

[0317]

[0318] Biological evaluation

[0319] Example 1 TYK2 JH2, TYK2 JH1, JAK1, JAK2, JAK3 kinase activity inhibition assay TYK2 JH2 kinase activity inhibition assay TYK2

[0320] TYK2 JH2 (N-His-Tev, 575-869) was expressed in Sf9, protein expression from our lab, fluorescein labeled probe was synthesized by our company, Anti-6xHis-terbium labled antibody was purchased from Cisbio. High purity Hepes, NaCl, MgCl2, DTT, BSA, Tween-20 and DMSO were purchased from Sigma.

[0321] The assay buffer used in the experiment was composed of 20 mM Hepes pH 7.5, 150 mM NaCl, 10 mM MgCl2, 2 mM DTT, 50 μg / mL BSA, and 0.015% Tween-20. The DMSO stock solution of the test compound was prepared, and according to the experimental requirements, DMSO was used for 12-point three-fold concentration gradient dilution. The 4% DMSO compound, TYK2 JH2 enzyme, fluorescein labeled probe, Anti-6xHis-terbium labled antibody were prepared with assay buffer, after preparation, 5 μL 4% DMSO compound, 5 μL TYK2 JH2 enzyme, 5 μL fluorescein labeled probe and 5 μL Anti-6xHis-terbium labled antibody were added into OptiPlate-384 White Opaque plate respectively, covered with film, 800 rpm for 1 min, incubated at room temperature for 1.5 h. The final concentrations of 4% DMSO compound, TYK2 JH2 enzyme, fluorescein labeled probe and Anti-6xHis-terbium labled antibody were 1%, 2.5 nM, 50 nM, 1x respectively. After 1.5 h, the plate was read on the SPARK multi-template reader of TECAN (Switzerland), the excitation wavelength was 340 nm, and the emission wavelength was 520 nm and 485 nm respectively. The IC50 value of the inhibitor was obtained by using Prism 8 (La Jolla, CA). 50

[0322] JH1, JAK1, JAK2, JAK3 kinase activity inhibition assay ​

[0323] TYK2 JH1 (NP_003322.3), JAK2 (NP_004963.1) and JA3 (NP_000206.2) were purchased from carnabio, JAK1 (N-GST-his-TEV, 850-1154) was expressed in HI5, protein expression from our lab, TK kit was purchased from Cisbio. High purity ATP, MgCl2, MnCl2, DTT and DMSO were purchased from Sigma. ​

[0324] The assay buffer used for TYK2 JH1 experiments consisted of 5 mM MgCl2, 1 mM MnCl2, 1 mM DTT, 12.5 μΜ SEB and 1x Enzymatic buffer.

[0325] The assay buffer used for JAK2 / 3 experiments consisted of 5 mM MgCl2, 1 mM DTT and 1x Enzymatic buffer.

[0326] The assay buffer used for JAK1 experiments consisted of 5 mM MgCl2, 1 mM MnCl2, 1 mM DTT and 1x Enzymatic buffer.

[0327] The DMSO stock solution of the test compound was prepared and diluted in DMSO in a three-fold concentration gradient of 12 points according to the experimental requirements. The 4% DMSO compound, enzyme, TK-Substrate, ATP were prepared using assay buffer. After preparation, 2.5 μΐ^ of 4% DMSO compound, 2.5 μΐ^ of enzyme, 5 μΐ^ of TK-Substrate / ATP mixture were added to OptiPlate-384 White Opaque plates, covered with film, 800 rpm for 1 min, and incubated at room temperature for 1 h. The final concentration of 4% DMSO compound was 1%, the final concentration of TYK2 JH1, JAK1, JAK2 and JAK3 enzymes was 0.2 ng / μΐ^, 10 ng / μΐ^, 0.125 ng / μΐ^ and 0.3 ng / μΐ^, respectively, and the final concentration of TK-Substrate / ATP was 0.25 μΜ and 3 μΜ, respectively. After 1 h, 5 μΐ^ of TK-Antibody-Cryptate antibody and Streptavidin-XL665 diluted in HTRF detection buffer were added, respectively, and incubated at room temperature for 1 h. The final concentration was 1x and 15.61 nM, respectively. After 1 h, the plates were read on a TECAN (Switzerland) SPARK multimode reader, with excitation wavelength at 320 nm and emission wavelength at 665 nm and 620 nm, respectively. The IC 50 values were obtained using Prism 8 (La Jolla, CA).

[0328] The IC 50 values of the representative compounds of the present application in TYK2 JH2, TYK2 JH1, JAK1, JAK2, JAK3 kinase activity inhibition assays are shown in Table 1.

[0329] Table 1

[0330]

[0331]

[0332]

[0333]

[0334] Note: Blank means not tested.

[0335] The results show that the representative compounds of the present application can effectively inhibit the kinase activity of TYK2 JH2, and show good selectivity to TYK2 JH1, JAK1, JAK2, JAK3.

[0336] Experimental Example 2 Permeability determination of Caco-2 cell monolayer

[0337] Materials:

[0338] Caco-2 cells and reagent solvents for the experiment were obtained commercially

[0339] Control Compound 1 and Control Compound 2 were prepared according to the method in international patent publication WO2019178079A1.

[0340] General method for permeability determination of Caco-2 cell monolayer:

[0341] 1. Preheating: HBSS buffer 37°C water bath preheating.

[0342] 2. Take the sample out from -20°C, ultrasonic treatment for not less than 1 minute.

[0343] 3. Buffer preparation

[0344] Blank solution on the dosing side:

[0345] A-to-B direction:

[0346] HBSS+ containing 0.3% DMSO and 5μM Lucifurin: 150μL DMSO and 125μL of 2mM Lucifurin solution were added to 50mL of HBSS+ buffer (pH 7.4).

[0347] HBSS+ containing 0.1% DMSO and 5μM Lucifurin: 50μL DMSO and 125μL of 2mM Lucifurin solution were added to 50mL of HBSS+ buffer (pH 7.4).

[0348] B-to-A direction:

[0349] HBSS+ containing 0.3% DMSO: 150μL DMSO was added to 50mL of HBSS+ buffer (pH 7.4).

[0350] HBSS+ with 0.1% DMSO: 50 μL DMSO was added to 50 mL of HBSS+ buffer (pH 7.4).

[0351] Receiving side solution:

[0352] A-to-B direction:

[0353] HBSS+ with 0.4% DMSO: 200 μL DMSO was added to 50 mL of HBSS+ buffer (pH 7.4).

[0354] B-to-A direction:

[0355] HBSS+ with 0.4% DMSO and 5 μM Lucifugum: 200 μL DMSO and 50 μL of 5 mM Lucifugum solution was added to 50 mL of HBSS+ buffer (pH 7.4).

[0356] 4. Measure the transmembrane resistance:

[0357] Remove the cell culture plates from the incubator and rinse the cells twice with HBSS buffer (400 μL per well for the upper cell plate and 25 mL for the lower support plate), and measure the transmembrane resistance at room temperature using a Millicell ERS resistance meter.

[0358] 5. Centrifuge the dosing side solution at 4000 rpm for 5 minutes. Collect the supernatant as the dosing side solution.

[0359] 6. Dosing:

[0360] A-B (dosing side): 600 μL A-to-B dosing solution (100 μL for Lucifugum culture start sample collection, 100 μL for culture start sample collection).

[0361] A-B (receiving side): 800 μL HBSS with 0.4% DMSO + .

[0362] B-A (dosing side): 900 μL B-to-A dosing solution (100 μL for culture start sample collection).

[0363] B-A (receiving side): 500 μL HBSS with 0.4% DMSO and 5 μM Lucifugum + (100 μL for Lucifugum culture start sample collection).

[0364] 7. The dosed upper and lower plates are placed in a 37°C incubator for 5 minutes to warm up. Then 100 μL is removed from the dosing side for the start time sample collection (A-to-B DO, B-to-A DO) and 100 μL is removed from each well of the upper plate and placed in a 96-well black fluorescence measuring plate for luciferin start time sample collection (DO).

[0365] 8. The upper and lower plates are brought together to start the permeability experiment and incubated for 90 minutes at 37°C.

[0366] 9. After the incubation period, the upper and lower plates are separated and 100 μL is removed from the dosing side and placed in a 96-well black fluorescence measuring plate and the amount of luciferin is measured after 90 minutes (excitation wavelength 485 nm, emission wavelength 535 nm).

[0367] 10. The dosing and receiving solution samples are diluted with 0.4% DMSO in HBSS buffer and then mixed with CAN containing an internal standard and sent for LC / MS analysis.

[0368] 11. Data Processing

[0369] Trans Epithelial Electrical Resistance (TEER) = (Resistance Value Sample - Resistance Value Blank) x Membrane Area

[0370] Luciferin Permeability:

[0371] Apparent Permeability Coefficient (P app ) = (Receiving Side Volume / (Membrane Area x Incubation Time)) x (Receiving Side Fluorescence Value at End of Incubation - Blank Solution Fluorescence Value) / ((Dosing Side Fluorescence Value at Start of Incubation - Blank Solution Fluorescence Value) x Dilution Factor)

[0372] Test Compound Permeability:

[0373] Apparent Permeability Coefficient (P app ) = (Receiving Side Volume / (Membrane Area x Incubation Time)) x (Receiving Side Drug Concentration at End of Incubation / (Dosing Side Drug Concentration at Start of Incubation x Dilution Factor)

[0374] This experiment uses Millipore Cell Culture Plates (PSHT 010R5): Membrane Area = 0.7 cm 2 , Receiving Side Volume = 0.8 mL (A-to-B) or 0.4 mL (B-to-A), Incubation Time = 90 minutes.

[0375] Compound Recovery:

[0376] % Recovery = 100 x (Total Compound at 90 minutes Dosing Side + Total Compound at 90 minutes Receiving Side) / (Total Compound at 0 minutes Dosing Side).

[0377] The apparent permeability coefficients of representative compounds of the present application, control compound 1 and control compound 2 are shown in Table 2.

[0378] Table 2

[0379]

[0380]

[0381] The results show that the ratio of the apparent permeability coefficients B-A / A-B of the compounds of the present application is lower than that of the control compounds, the drug absorption capacity is better than that of the control compounds, and the efflux is less likely to occur, and the drug efficacy in the cell is more likely to occur.

Claims

1. Compounds represented by general formula (Ic): Or its tautomers, meso compounds, racemates, enantiomers, diastereomers, or pharmaceutically acceptable salts thereof, wherein... A3 is selected from C or N; R1 is selected from -CH3, R2 is a C1-C6 alkyl group; R3 is -OR a ; R4' is a C1-C6 alkyl group or -NH2; Each R a Independently selected from hydrogen, C1-C6 alkyl, cyano or -C(O)R c ; Each R c Independently selected from halogenated C1-C6 alkyl groups; and n is 1.

2. The compound according to claim 1, wherein, R2 is -CH3.

3. The compound according to claim 1, wherein, R3 is 4. The compound according to claim 1, wherein, R4' is either -CH3 or -NH2.

5. The compound of formula (I) according to claim 1, wherein the compound is selected from:

6. A pharmaceutical composition comprising a therapeutically effective amount of the compound of any one of claims 1-5, an isomer thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient.

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