CN116249526A - TYK2 selective inhibitors and uses thereof - Google Patents

Abstract

The application discloses TYK2 selective inhibitor and application thereof, in particular to a compound shown in a formula (I) or a tautomer, a meso form, a racemate, an enantiomer, a diastereoisomer or pharmaceutically acceptable salt thereof, and application thereof in preparing medicines for treating TYK 2-mediated diseases.

Description

TYK2 selective inhibitors and uses thereof technical field

The application belongs to the field of chemical medicine, and specifically relates to a TYK2 selective inhibitor and its application.

Background technique

Autoimmune disease is a family of at least 80 diseases, such as rheumatoid arthritis, systemic lupus erythematosus, and inflammatory bowel disease, is a group of immune cell activation and excessive production of autoantibodies that mistakenly attack their own organs, tissues and cellular diseases. Autoimmune diseases affect 5%-10% of people worldwide (Shoenfeld Y, Tincani A, Gershwin ME (2012) Sex gender and autoimmunity. J Autoimmun 38:J71–J73). Autoimmune diseases, as chronic and debilitating diseases with high medical costs and reduced quality of life for patients, have become a huge burden on patients, their families and society. Although the pathogenesis of these diseases is not fully understood, studies have shown that multiple factors such as genetics, environment and immune response play an important role in the development of the disease.

Kinases play a very important role in regulating immune cell functions (Deng, Bellanti, Zheng. Essential Kinases and Transcriptional Regulators and Their Roles in Autoimmunity [J]. Biomolecules, 2019, 9(4)). Members of the Janus kinase (JAK) family include JAK1, JAK2, JAK3, and TYK2, and 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 A V , 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 tyrosine kinase 2 Other variants of (TYK2) are associated with autoimmune diseases such as Crohn's disease, psoriasis, systemic lupus erythematosus, and rheumatoid arthritis, further suggesting 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 T J. Association of polymorphisms across the tyrosine kinase gene, TYK2 in UKSLE 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 regulates signal transduction cascades in IL-12, IL-23 and type I interferon receptors It is very important in the downstream (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 can promote the differentiation and proliferation of Th1 and Th17. Human genomics studies found a strong association between IL-12R and IL-23B (encoding p40 subunit) polymorphisms and 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 systems, 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 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 interferon have therapeutic benefit in human autoimmune diseases.

Studies have shown that selective inhibition of TYK2 activity can be used as a new method for the treatment of various autoimmune diseases that can balance the relationship between therapy and safety (Leitner, Nicole, R, et al..Tyrosine kinase 2-Surveillant of tumours and bona fide oncogene [J]. Cytokine, 2017.). TYK2, as well as other members of the JAK family, is characterized by a dual kinase domain, a tyrosine kinase domain (JH1) and a pseudokinase domain (JH2), respectively. 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.), JAKs Partial mutations in JH2 have been shown to be associated with hematological and immunological diseases. Therefore, TYK2 JH2 selective inhibitors may be able to inhibit TYK2 activity more specifically.

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 interferon in vivo.

Contents of the invention

The application discloses a compound that can be used as a selective inhibitor of TYK2 and its use in the preparation of drugs for preventing or treating related diseases mediated by TYK2.

On the one hand, the application discloses a compound represented by general formula (I):

or its tautomers, mesoforms, racemates, enantiomers, diastereoisomers or pharmaceutically acceptable salts thereof,

且当A ₂为N时，A ₄、A ₅、A ₆、A ₇、A ₈中至少有1个为N； Wherein, A ₁ , A ₂ , A ₄ , A ₅ , A ₆ , A ₇ , A ₈ are selected from C or N, A ₃ is selected from C, N or

And when A ₂ is N, at least one of A ₄ , A ₅ , A ₆ , A ₇ and A ₈ is N;

为饱和或不饱和环；

is a saturated or unsaturated ring;

R ₁ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

R ₂ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

R ₄ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

Each R _a and R _b are independently selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₃ - C ₆ heterocycloalkyl, halogen, hydroxyl, cyano, nitro, -C(O)NR _c R _d , -C(O)R _c , -(CH ₂ ) _n C(O)OR _c , -OR _c , -(CH ₂ ) _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) ₂ R _c , the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl is optionally substituted by 1-3 R _c ;

Each R _c and R _d are independently selected from hydrogen, halogen, carbonyl, -C(O)CH ₃ , hydroxyl, cyano, nitro, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkane radical, C ₃ -C ₆ cycloalkyl or halogenated C ₃ -C ₆ cycloalkyl; and

Each n is independently selected from 0, 1, 2 or 3.

In some embodiments, the compound represented by general formula (Ia):

or its tautomers, mesoforms, racemates, enantiomers, diastereoisomers or pharmaceutically acceptable salts thereof,

且当A ₂为N时，A ₄、A ₅、A ₆、A ₇、A ₈中至少有1个为N； Wherein, A ₁ , A ₂ , A ₄ , A ₅ , A ₆ , A ₇ , A ₈ are selected from C or N, A ₃ is selected from C, N or

And when A ₂ is N, at least one of A ₄ , A ₅ , A ₆ , A ₇ and A ₈ is N;

为饱和或不饱和环；

is a saturated or unsaturated ring;

R ₁ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

R ₂ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

R ₃ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

R ₄ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

Each R _a and R _b are independently selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₃ - C ₆ heterocycloalkyl, halogen, hydroxyl, cyano, nitro, -C(O)NR _c R _d , -C(O)R _c , -(CH ₂ ) _n C(O)OR _c , -OR _c , -(CH ₂ ) _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) ₂ R _c , the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl is optionally substituted by 1-3 R _c ;

Each R _c and R _d are independently selected from hydrogen, halogen, carbonyl, -C(O)CH ₃ , hydroxyl, cyano, nitro, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkane radical, C ₃ -C ₆ cycloalkyl or halogenated C ₃ -C ₆ cycloalkyl; and

Each n is independently selected from 0, 1, 2 or 3.

In some embodiments, general formula (I) is a compound represented by general formula (Ib):

Or its tautomers, mesomers, racemates, enantiomers, diastereoisomers or pharmaceutically acceptable salts thereof, wherein A ₁ and A ₃ are selected from C or N ;

R ₁ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

R ₂ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

R ₃ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, halogenated C ₁ -C ₆ alkyl, halogenated C ₂ -C ₆ alkenyl, Halogenated C ₂ -C ₆ alkynyl, halogen, cyano, nitro, -C(O)NR _a R _b , -C(O)R _a , -C(O)OR _a , -OR _a , -( CH ₂ ) _n OR _a , -NR _a R _b or -S(O) ₂ R _a ;

R ₄ ' is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, halogenated C ₁ -C ₆ alkyl, halogenated C ₂ -C ₆ alkenyl , halogenated C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, halogenated C ₃ -C ₆ cycloalkyl, halogen, cyano, nitro or -NR _a R _b ;

Each R _a and R _b are independently selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₃ - C ₆ heterocycloalkyl, halogen, hydroxyl, cyano, nitro, -C(O)NR _c R _d , -C(O)R _c , -(CH ₂ ) _n C(O)OR _c , -OR _c , -(CH ₂ ) _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) ₂ R _c , the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl is optionally substituted by 1-3 R _c ;

Each R _c and R _d are independently selected from hydrogen, halogen, carbonyl, -C(O)CH ₃ , hydroxyl, cyano, nitro, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkane radical, C ₃ -C ₆ cycloalkyl or halogenated C ₃ -C ₆ cycloalkyl; and

Each n is independently selected from 0, 1, 2 or 3.

In other embodiments, general formula (I) is a compound represented by general formula (Ic):

Or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt thereof, wherein, _A3 is selected from C or N;

R ₁ is selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ 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) ₂ R _a or a 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms, the alkyl, alkenyl, alkynyl, A 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms is optionally substituted by 1-3 R _a ;

R ₂ is selected from hydrogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, halogen, cyano, -C(O)OR _a , -(CH ₂ ) _n OR _a or -NH ₂ ;

R ₃ is selected from hydrogen, C ₁ -C ₆ alkyl or -OR _a ;

R ₄ ' is selected from hydrogen, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, halogen or -NH ₂ ;

Each R _a and R _b are independently selected from hydrogen, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cycloalkyl, C ₃ - C ₆ heterocycloalkyl, halogen, hydroxyl, cyano, nitro, -C(O)NR _c R _d , -C(O)R _c , -(CH ₂ ) _n C(O)OR _c , -OR _c , -(CH ₂ ) _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) ₂ R _c , the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl is optionally substituted by 1-3 R _c ;

Each R _c and R _d are independently selected from hydrogen, halogen, carbonyl, -C(O)CH ₃ , hydroxyl, cyano, nitro, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkane radical, C ₃ -C ₆ cycloalkyl or halogenated C ₃ -C ₆ cycloalkyl; and

Each n is independently selected from 0, 1, 2 or 3.

In other embodiments, R ₁ is selected from hydrogen, -CH ₃ ,

In other embodiments, R ₂ is selected from hydrogen, —CH ₃ ,

In other embodiments, _R3 is

或-C(O)NH ₂。 In other embodiments, R ₄ is hydrogen, -C(O)CH ₃ ,

or -C(O) _NH2 .

In other embodiments, the compound represented by general formula (I) is selected from:

On the other hand, the present application also provides the use of the aforementioned compounds, their isomers or pharmaceutically acceptable salts thereof in the preparation of drugs for diseases mediated by TYK2.

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 transplant-related disease.

In other embodiments, the disease is an autoimmune disease.

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

In other embodiments, the disease is an inflammatory disease.

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.

On the other hand, the present application also provides a pharmaceutical composition, which comprises a therapeutically effective amount of the aforementioned compound, its isomer or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient.

the term:

Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

The term "isomer" includes enantiomeric, diastereomeric and geometric (or conformational) isomeric forms of a given structure. For example, the present application includes R and S configurations, Z and E double bond isomers, Z and E conformational isomers, single stereochemical isomers and enantiomers, diastereomers for each asymmetric center. Mixtures of isomers and geometric (or conformational) isomers.

The term "pharmaceutically acceptable salt" refers to, for example, acid addition and/or base salts thereof. Suitable acid addition salts are formed with acids which form non-toxic salts, eg hydrochlorides/chlorides. Suitable base salts are formed from bases which form non-toxic salts, such as calcium and sodium salts. Half-salts of acids and bases, such as the hemisulfate and hemicalcium salts, may also be formed.

The term "therapeutically effective amount" refers to the amount of a compound of the present application that (i) treats a specific disease, condition or disorder; (ii) alleviates, relieves or eliminates one or more symptoms of a specific disease, condition or disorder or (iii) preventing or delaying the onset of one or more symptoms of a particular disease, condition or disorder described herein.

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 formulated therewith.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably 1 to 6 carbon atoms atom of the alkyl group. Non-limiting examples of lower alkyl groups containing 1 to 6 carbon atoms include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl base, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methyl Butyl, 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, etc.

The term "alkenyl" refers to an aliphatic hydrocarbon having at least one carbon-carbon double bond, including straight and branched chains having at least one carbon-carbon double bond. In some embodiments, an alkenyl group has 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 " _C2-6 alkenyl" includes straight or branched chain unsaturated groups (having at least one carbon-carbon double bond) of 2 to 6 carbon atoms, including but not limited to ethenyl, 1-propenyl , 2-propenyl (allyl), isopropenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, etc.

The term "alkynyl" refers to an aliphatic hydrocarbon having at least one carbon-carbon triple bond, including straight and branched chains having at least one carbon-carbon triple bond. In some embodiments, an alkynyl group has 2 to 20, 2 to 10, 2 to 6, or 3 to 6 carbon atoms. For example, " _C2-6 alkynyl" includes straight or branched chain alkynyl groups as defined above having 2 to 6 carbon atoms.

The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring containing 3 to 20 carbon atoms, preferably containing 3 to 12 carbon atoms, more preferably containing 3 to 6 carbon atoms (for example 3, 4, 5 or 6 carbon atoms), most preferably contain 5 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene Base, cyclooctyl, etc.; polycyclic cycloalkyl includes spiro ring, fused ring and bridged ring cycloalkyl.

The term "spirocycloalkyl" refers to a polycyclic group of 5 to 20 membered monocyclic rings sharing one carbon atom (called a spiro atom), which may contain one or more double bonds, but none of the rings has complete conjugation The π-electron system. Preferably it is 6 to 14 yuan, more preferably 7 to 10 yuan (eg 7, 8, 9 or 10 yuan). According to the number of spiro atoms shared between the rings, the spirocycloalkyl group can be divided into single spirocycloalkyl, double spirocycloalkyl or polyspirocycloalkyl, preferably single spirocycloalkyl and double spirocycloalkyl. 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 monospirocycloalkyl group.

The term "fused cycloalkyl" refers to a 5 to 20 membered all-carbon polycyclic group in which each ring of the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more rings may contain one or Multiple double bonds, but none of the rings have a fully conjugated π-electron system. Preferably it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic condensed cycloalkyl groups, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicycloalkyl groups.

The term "bridged cycloalkyl" refers to a 5 to 20 membered, all-carbon polycyclic group having any two rings sharing two carbon atoms not directly attached, which may contain one or more double bonds, but none of the rings has a complete Conjugated π-electron systems. Preferably it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic.

The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O ) m (where m is an integer from 0 to 2), but excluding ring portions of -O-O-, -O-S- or -S-S-, the remaining ring atoms being carbon. Preferably contain 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably contain 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; most preferably contain 5 to 6 ring atoms, of which 1 to 2 or 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrazolyl, Hydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc., preferably tetrahydropyranyl, piperidinyl, pyrrolidinyl. Polycyclic heterocycloalkyls include spiroheterocyclyls, fused heterocyclyls and bridged heterocyclyls.

The term "spiroheterocyclyl" refers to a polycyclic heterocyclic group that shares one atom (called a spiro atom) between 5 to 20-membered monocyclic rings, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O )m (wherein m is an integer from 0 to 2), the remaining ring atoms are carbon. It may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system. Preferably it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiro atoms shared between the rings, the spiroheterocyclyl can be divided into single spiroheterocyclyl, double spiroheterocyclyl or polyspiroheterocyclyl, preferably single spiroheterocyclyl and double spiroheterocyclyl. 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 monospiro heterocyclic group.

The term "fused heterocyclyl" refers to a 5 to 20 membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system, and one or more rings may contain one or more double bond, but none of the rings has a fully conjugated π-electron system, where 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 The atom is carbon. Preferably it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic groups.

The term "bridged heterocyclyl" refers to a 5 to 14 membered polycyclic heterocyclic group in which any two rings share two atoms not directly attached, which may contain one or more double bonds, but none of the rings has a complete shared bond. A pi-electron system of a yoke 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 it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic.

The term "aryl" refers to a 6 to 14 membered all-carbon monocyclic or fused polycyclic (that is, rings sharing adjacent pairs of carbon atoms) group, preferably 6 to 10 membered, having a conjugated pi-electron system, such as benzene base and naphthyl.

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; preferred examples are imidazolyl, furyl, thienyl, thiazolyl, pyryl Azolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl and pyridazinyl, etc.

The term "hydroxyalkyl" refers to an alkyl group substituted by a hydroxy group, wherein alkyl is as defined above.

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

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

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

The term "deuterated alkoxy" refers to an alkoxy group substituted with one or more deuterium atoms, wherein alkoxy group is as defined above.

The term "cycloalkylalkyl" refers to an alkyl group substituted by one or more cycloalkyl groups, wherein cycloalkyl and alkyl are as defined above.

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

The term "heterocyclylalkyl" refers to an alkyl group substituted by one or more heterocyclyl groups, wherein heterocyclyl and alkyl are as defined above.

The term "arylalkyl" refers to an alkyl group substituted with one or more aryl groups, wherein aryl and alkyl are as defined above.

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

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

The term "amino" refers to _-NH2 .

The term "cyano" refers to -CN.

The term "nitro" refers to _-NO2 .

The term "carboxy" refers to -C(O)OH.

Detailed ways

Intermediate preparation

Preparation of Intermediate 1.1: (R)-2-Bromo-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine

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

Step 2: 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 .), CH ₃ I (17.5g, 124mmol, 1.1eq.), reacted at 25°C for 1.5h. The reaction solution was poured into 400mL ice water, extracted with EA (100mL*3), the crude product was purified by column chromatography (PE/EtOAc=5:1-3:1), and then chirally resolved to obtain the title compound (RT=2.26 min, 12g, yield 40%). MS (m/z) = 272.02 [M+H] ⁺ .

Chiral resolution conditions:

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

Mobile phase: A is _CO2 and B is ethanol (0.1% _NH3H2O ); Gradient: B 15%; Flow rate: 70mL/ _min ; Back pressure: 100bar; Column temperature: 35°C; Wavelength: 254nm; Cycle time : ~ 1.5min; interval time: 0.5min; sample preparation: the compound was dissolved in 200mL ethanol.

Intermediate 1.2-1.6 was prepared by referring to the method of Intermediate 1.1.

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

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

Step 2: Preparation of 6-chloro-3-vinyl-1H-pyrrolo[3,2-c]pyridine. 3-bromo-6-chloro-1H-pyrrolo[3,2-c]pyridine (1.00g, 4.32mmol, 1eq.), 4,4,5,5-tetramethyl-2-vinyl-1, 3,2-dioxaborane (665mg, 4.32mmol, 1eq.), Pd(dppf)Cl ₂ (3.16g, 4.32mmol, 1eq.), K ₂ CO ₃ (597mg, 4.32mmol, 1eq.) In water (2 mL) and dioxane (10 mL), react at 100° C. for 3 h under nitrogen protection. The reaction solution was spin-dried, 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] ⁺ .

Intermediates 2.2-2.9 were prepared by referring to the method of Intermediate 2.1.

Preparation of intermediate 3: Preparation of 6-chloro-3-cyclopropyl-1H-pyrrolo[3,2-c]pyridine

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

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

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

Step 4: Preparation of 6-chloro-3-cyclopropyl-1H-pyrrolo[3,2-c]pyridine. 6-Chloro-3-cyclopropyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridine (0.15g, 464μmol, 1eq.) was dissolved in 10mL of tetrabutylammonium fluoride in THF (1M) solution, and refluxed for 10h. Water was added and extracted with EA, the crude product was purified by column chromatography (PE/EtOAc=10:1-1:1) to obtain the title compound (0.10 g, yield 94%). MS (m/z) = 193.05 [M+H] ⁺ .

Preparation of Intermediate 4: 6-Chloro-3-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrrolo[3,2-c]pyridine

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

Preparation of Intermediate 5: 3-(6-Chloro-1H-pyrrolo[3,2-c]pyridin-3-yl)-1-methylazetidin-3-ol

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

Synthesis of Intermediate 6: 3-(6-Chloro-1H-pyrazolo[4,3-c]pyridin-3-yl)bicyclo[1.1.1]pentane-1-carbonitrile

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 (2.00g, 14.6mmol, 1eq.), N,O-dimethylhydroxylamine hydrochloride (1.42g, 14.6mmol, 1eq.), HATU (6.65g, 17.5mmol, 1.20eq.), DIEA (3.77g, 29.2mmol, 2.00eq.) were dissolved in DCM (20mL), and reacted at 20°C for 12h. Add 10 mL of citric acid to the reaction solution, separate and wash once with 10 mL of saturated sodium carbonate, then once with 10 mL of saturated sodium chloride, and concentrate to obtain the title compound (2 g, yield 76%).

Step 2: Preparation of 3-(4,6-dichloronicotinoyl)bicyclo[1.1.1]pentane-1-carbonitrile. 5-Bromo-2,4-dichloropyridine (2.10g, 9.25mmol, 1eq.) was dissolved in THF (20mL), and isopropylmagnesium chloride (10.2mmol, 5.09mL, 1.1eq.) was added dropwise at -78°C, Reacted for 2h, then added dropwise 3-cyano-N-methoxy-N-methylbicyclo[1.1.1]pentane-1-carboxamide (2.00g, 11.10mmol, 1.20eq.) at -78°C, naturally Rising to room temperature, reaction 10h. 20 mL of saturated ammonium chloride solution was added to the reaction solution, separated and concentrated. The crude product was purified by column chromatography (PE/EtOAc=10:1-1:1) to obtain the title compound (1.5 g, yield 60%). MS (m/z) = 267.00 [M+H] ⁺ .

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

Synthesis of intermediate 7: tert-butyl 3-(6-chloro-1H-pyrrolo[3,2-c]pyridin-3-yl)azetidine-1-carboxylate

Step 1: 3-bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3, 2-c]pyridine. MS (m/z) = 361.01 [M+H] ⁺ .

Step 2: 3-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)tert-butyl-1-pyrrolo[3,2-c]pyridine-3- base)-3-hydroxyazetidine-1-carboxylate. 3-Bromo-6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridine (5.00g, 13.8mmol, 1eq .) was dissolved in THF (30mL), added dropwise n-BuLi (14.5mmol, 3.20mL, 1.05eq.) at -78°C, kept at -78°C for 1h, and then added dropwise 3-oxoazetidine- 1-tert-butyl carboxylate (2.37g, 13.8mmol, 1eq.), react at -78°C for 1h, quench the reaction solution with saturated ammonium chloride, and purify the crude product by column chromatography (PE/EtOAc=10:1- 2:1). The title compound (3.50 g, 55.8% yield) was obtained. MS (m/z) = 454.19 [M+H] ⁺ .

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

Step 4: 3-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1-pyrrolo[3,2-c]pyridin-3-yl)nitrogen Preparation of tert-butyl heterocyclobutane-1-carboxylate. 3-(6-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridin-3-yl)-3-( Preparation of tert-butyl ((methylthio)carbonthio)oxy)azetidine-1-carboxylate (3.00 g, 5.51 mmol, 1 eq.) was dissolved in toluene (60 mL) and AIBN (90.5 mg , 551μmol, 0.1eq.), Bu ₃ SnH (2.41g, 8.27mmol, 1.5eq.), reacted at 85°C for 3h under nitrogen protection. The crude product was purified by column chromatography (PE/EtOAc=10:1-3:1) to obtain a colorless liquid (2.0 g, yield 82%). MS (m/z) = 438.19 [M+H] ⁺ .

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

Synthesis of intermediate 8: tert-butyl 6-(6-chloro-1H-pyrrolo[3,2-c]pyridin-3-yl)-2-azaspiro[3.3]heptane-2-carboxylate

Intermediate 8 was prepared by referring to the method of Intermediate 7. MS (m/z) = 348.14 [M+H] ⁺ .

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

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

Step 2: Preparation of 6-chloro-3-vinyl-1H-pyrrolo[3,2-c]pyridine. 3-bromo-6-chloro-1H-pyrrolo[3,2-c]pyridine (1.00g, 4.32mmol, 1eq.), 4,4,5,5-tetramethyl-2-vinyl-1, 3,2-dioxaborane (665mg, 4.32mmol, 1eq.), Pd(dppf)Cl ₂ (3.16g, 4.32mmol, 1eq.), K ₂ CO ₃ (597mg, 4.32mmol, 1eq.) In water (2 mL) and dioxane (10 mL), react at 100° C. for 3 h under nitrogen protection. The reaction solution was spin-dried, 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] ⁺ .

Step 3: 6-chloro-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-vinyl-1H-pyrrolo[3,2- c] Preparation of pyridines. 2-Bromo-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine (152mg, 559μmol, 1eq.), 6-chloro-3-vinyl-1H-pyrrolo[3,2 -c] pyridine (0.1g, 559μmol, 1eq.), K ₂ CO ₃ (232mg, 1.68mmol, 3.0eq.), CuI (5.33mg, 27.99μmol, 0.05eq.), N,N-dimethylethyl Diamine (4.94 mg, 56 μmol, 0.1 eq.) was dissolved in 1,4-dioxane (30 mL), and reacted under reflux for 12 h under nitrogen protection. The reaction solution was spin-dried, 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] ⁺ .

Step 4: N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-vinyl-1H-pyrrolo[3,2-c ]pyridin-6-yl)acetamide preparation. 6-Chloro-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-vinyl-1H-pyrrolo[3,2-c]pyridine (0.20g, 540μmol, 1eq.), acetamide (47.9mg, 811μmol, 1.5eq.), Cs ₂ CO ₃ (528mg, 1.62mmol, 3.0eq.), Pd ₂ (dba) ₃ (49.5mg, 54.1μmol , 0.1eq.), X-phos (52mg, 108μmol, 0.2eq.) was dissolved in 1,4-dioxane (30mL), and refluxed for 12h under nitrogen protection. The reaction solution was spin-dried, and the crude product was purified by column chromatography (PE/EtOAc=2:1-0:1) to obtain the title compound (0.2 g, yield 94%). MS (m/z) = 393.18 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.45(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).

The compounds of Examples 2-20 were prepared with reference to the method of Example 1.

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

N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-vinyl-1H-pyrrolo[3,2-c]pyridine- 6-yl)acetamide (0.05g, 127μmol, 1eq.) was dissolved in MeOH (10mL), replaced with hydrogen three times, and reacted at 20°C for 12h. 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 H NMR (400MHz,DMSO)δ10.38(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).

The compounds of Examples 22-27 were prepared by referring to the method of Example 21.

Example 28a or Example 28b: N-(3-((1s,3s)-3-cyanocyclobutyl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methanol Basepyridin-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

Step 1: Preparation of tert-butyl 3-bromo-6-chloro-1H-pyrrolo[3,2-c]pyridine-1-carboxylate. 3-Bromo-6-chloro-1H-pyrrolo[3,2-c]pyridine (2.00g, 8.64mmol, 1eq.), (Boc) _2O (1.89g, 8.64mmol, 1.0eq.), DMAP ( 105mg, 864μmol, 0.1eq.), TEA (1.31g, 12.9mmol, 1.5eq.) was dissolved in THF (20mL), and reacted at 25°C for 1h. The crude product was purified by column chromatography (PE/EtOAc=10:1-3:1) to obtain the title compound (2.80 g, 97.7%). MS (m/z) = 330.98 [M+H] ⁺ .

Step 2: Preparation of tert-butyl 6-chloro-3-(3-cyano-1-hydroxycyclobutyl)-1H-pyrrolo[3,2-c]pyridine-1-carboxylate. 3-Bromo-6-chloro-1H-pyrrolo[3,2-c]pyridine-1-carboxylic acid tert-butyl ester (2.80g, 8.44mmol, 1eq.) was dissolved in THF (30mL) at -78°C Add n-BuLi (8.87mmol, 3.20mL, 1.05eq.) dropwise, keep at -78°C for 1h, then add 3-oxocyclobutanenitrile (883mg, 9.29mmol, 1.1eq.) dropwise, and react at -78°C for 1h. The reaction solution was quenched with saturated ammonium chloride, separated, dried, filtered and concentrated. The crude product was purified by column chromatography (PE/EtOAc=5:1-1:1) to obtain the title compound (2.00 g, 68.10%). MS (m/z) = 348.10 [M+H] ⁺ .

Step 3: Preparation of 3-(6-chloro-1H-pyrrolo[3,2-c]pyridin-3-yl)cyclobutane-1-carbonitrile. tert-butyl 6-chloro-3-(3-cyano-1-hydroxycyclobutyl)-1H-pyrrolo[3,2-c]pyridine-1-carboxylate (2.00g, 5.75mmol, 1eq.) , Et ₃ SiH (6.69g, 57.5mmol, 10.0eq.), TFA (6.56g, 57.5mmol, 10.0eq.) were dissolved in DCM (30mL), and reacted at 25°C for 1h. The reaction solution was spin-dried, and 10 mL of saturated sodium carbonate solution and 10 mL of ethyl acetate were added. After separation, the organic phase was dried, filtered and concentrated to obtain the title compound (1.00 g, 75.06%). MS (m/z) = 232.06 [M+H] ⁺ .

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

Step 5: N-(3-(3-cyanocyclobutyl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrole [3,2-c]pyridin-6-yl)acetamide preparation. 3-(6-chloro-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridine-3 -yl)cyclobutane-1-carbonitrile (1.00g, 2.36mmol, 1eq.), acetamide (209mg, 3.55mmol, 1.5eq.), _Cs2CO3 ( _2.31g , 7.09mmol, 3.0eq.), Pd ₂ (dba) ₃ (216mg, 236μmol, 0.1eq.), X-phos (225mg, 473μmol, 0.2eq.) were dissolved in 1,4-dioxane (50mL), and refluxed for 12h under nitrogen protection. The reaction solution was spin-dried, and the crude product was purified by column chromatography (EtOAc) to obtain the title compound (0.2 g, yield 19%). MS (m/z) = 446.21 [M+H] ⁺ .

Step 6: The compound obtained in Step 5 was purified by preparative TLC (developing solvent: EtOAc, R _f 1 = 0.43, R _f 2 = 0.36) to obtain the compounds of Example 28a and Example 28b. ¹ H NMR (400MHz,DMSO)δ10.38(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＝9.7Hz,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 (400MHz,DMSO)δ10.40(s,1H),9.07(s,1H),8.61(s,1H),7.99(s,1H),7.58(s,1H),7.30(s,1H ),4.21(d,J＝9.7Hz,1H),4.08–3.86(m,4H),3.53(ddd,1H),3.15(s,3H),2.87–2.75(m,2H),2.72–2.58( m,3H), 2.47(s,4H), 2.10(s,3H).

Example 29a and Example 29b were prepared in a similar manner with reference to Example 28a and Example 28b (developing solvent: EtOAc, R _f 1 = 0.43, R _f 2 = 0.36).

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

Step 1: (R)-2-(4-(6-Acetamido-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrole [3,2-c]pyridin-3-yl)cyclohexyl)ethyl acetate preparation. 2-(4-(6-acetamido-1-(6-((R)-3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[ 3,2-c]pyridin-3-yl)cyclohex-3-en-1-yl)ethyl acetate was prepared by the method similar to Example 1 from Intermediate 2.5 and Intermediate 1.1, and then similar to Example 22 method to obtain the title compound. MS (m/z) = 535.28 [M+H] ⁺ .

Step 2: (R)-2-(4-(6-Acetamido-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrole [3,2-c]pyridin-3-yl)cyclohexyl)acetic acid preparation. (R)-2-(4-(6-acetylamino-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3 ,2-c]pyridin-3-yl)cyclohexyl)ethyl acetate (0.30g, 561μmol, 1eq.), LiOH·H ₂ O (47.1mg, 1.12mmol, 2.0eq.) was dissolved in H ₂ O (3mL ) and THF (9mL), reacted at 20°C for 12h. The reaction solution was spun out of THF, and the remaining liquid was adjusted to pH 5 with 1N HCl. A large amount of solid precipitated, filtered, and the filter cake was dried. The title compound (0.2 g, yield 70%) was obtained. MS (m/z) = 507.25 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.38(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).

Example 31 was prepared in a similar manner with reference to Example 30.

Example 32: (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

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

Step 2: (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(piperidin-4-yl)- Preparation of 1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide. (R)-4-(6-acetamido-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2 -c] pyridin-3-yl)piperidine-1-carboxylate tert-butyl ester (500 mg, 909 μmol, 1 eq.) was dissolved in DCM (5 mL) and TFA (1 mL), and reacted at 20° C. for 1 h. The reaction solution was adjusted to pH 10 with saturated sodium carbonate, extracted by adding 20mL of EA, separated and spin-dried. The title compound (0.2 g, yield 49%) was obtained. MS (m/z) = 450.24 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.40(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).

Example 33 was prepared in a similar manner with reference to Example 32.

Example 34a or 34b: N-(3-((1s,4S)-4-aminocyclohexyl)-1-(6-((R)-3-methoxytetrahydrofuran-3-yl)-4-methan Basepyridin-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

Examples 34a and 34b were prepared from the compound of Example 33 by chiral resolution (RT ₁ =3.38min, RT ₂ =4.19min).

Chiral resolution conditions:

Instrument: Waters 200, preparative SFC (QC-R-LC-07); Chromatographic column: ChiralPak IC, 250×30mm ID, 5 μm; Mobile phase: A is CO ₂ and B is ethanol (0.1% NH ₃ H ₂ O) ; Gradient: B 30%; Flow rate: 70mL/min; Back pressure: 100bar; Column temperature: 35°C; Wavelength: 210nm;

¹ H NMR (400MHz,DMSO)δ10.37(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] ⁺ .

¹ H NMR (400MHz,DMSO)δ10.37(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] ⁺ .

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

(R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(piperidin-4-yl)-1H-pyrrole [3,2-c]pyridin-6-yl)acetamide (100 mg, 222 μmol, 1 eq.) was dissolved in 37% aqueous acetaldehyde (2.73 mL) and DCM (2.27 mL), stirred for 1 h, then added STAB ( 377mg, 445μmol, 2.0eq.), 20°C for 14h, the reaction solution was adjusted to pH 10 with saturated sodium carbonate, added 20mL EA for extraction, separated, dried the organic phase, filtered, and spin-dried to obtain the title compound (0.1g, yield 94%). MS (m/z) = 478.27 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.41(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).

Example 36a: (R)-N-(3-(azetidin-3-yl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine-2 -yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide and Example 36b: (R)-N-(1-(6-(3-methoxytetrahydrofuran-3- Base)-4-methylpyridin-2-yl)-3-(1-methylazetidin-3-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)B Amide

Referring to the methods of Example 1 and Example 35, the title compound was obtained.

36a: MS (m/z) = 422 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.39(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).

36b: MS (m/z) = 436.23 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.42(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).

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

Referring to the methods of Example 1 and Example 35, the title compound was obtained.

37a: MS (m/z) = 462 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.40(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).

37b: MS (m/z) = 476.26 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.35(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).

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

(R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(piperidin-4-yl)-1H-pyrrole [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 , 888μmol, 1.2eq.), after reacting at 20°C for 1h, the reaction solution was poured into 10mL of water, separated and spin-dried to obtain the title compound (0.3g, yield 82%). MS (m/z) = 492.25 [M+H] ⁺ . ¹ H NMR (400MHz, CDCl ₃ )δ8.94(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).

Examples 39-40 were prepared in a similar manner with reference to Example 38.

Example 41a or Example 41b: N-(7-((1s,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-((1r,3R)-3-cyano ring Butyl)-5-(6-((R)-3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-7H-pyrrolo[2,3-c]pyridazine -3-yl)acetamide

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

Step 2: Preparation of 3-cyanocyclobutyl methanesulfonate. 3-Hydroxycyclobutyronitrile (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), Reaction at 20°C for 2h. The reaction solution was poured into 10 mL of water, separated and concentrated to obtain the title compound (2.7 g, yield 99%).

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

Step 4: (R)-3-(3-chloro-5-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-7H-pyrrolo[2, Preparation of 3-c]pyridazin-7-yl)cyclobutane-1-carbonitrile. 3-(5-Bromo-3-chloro-7H-pyrrolo[2,3-c]pyridazin-7-yl)cyclobutane-1-carbonitrile (500mg, 1.60mmol, 1eq.), coupled with boronic acid Nacohol ester (407 mg, 1.60 mmol, 1.0 eq.), Pd(dppf)Cl ₂ DCM (131 mg, 160 μmol, 0.1 eq.), potassium acetate (315 mg, 3.21 mmol, 2.0 eq.) were dissolved in 10 mL of dioxane , reacted at 110°C for 2h under the protection of nitrogen, and the reaction was cooled to room temperature. -3-yl)-4-methyl-pyridine (437mg, 1.60mmol, 1eq) and water (2mL), continued to react at 110°C for 1h. The reaction solution was directly made into sand, and the crude product was purified by column chromatography (PE/EtOAc=2:1-1:1) to obtain an off-white solid as the title compound (0.5 g, yield 74%). MS (m/z) = 424.15 [M+H] ⁺ .

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

Step 6: The compound obtained in Step 5 was purified by preparative TLC (developing solvent: EtOAc, R _f 1 = 0.45, R _f 2 = 0.42) to obtain the compounds of Example 41a and Example 41b. ¹ H NMR (400MHz,DMSO)δ10.93(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 H NMR (400MHz, DMSO) δ10.93(s,1H),9.35(s,1H),9.11(s,1H),7.78(s,1H),7.20(s,1H),5.58–5.47(m ,1H),4.21(d,1H),4.08–3.97(m,2H),3.89(d,1H),3.41–3.35(m,1H),3.11(s,3H),3.08–3.00(m,2H ), 2.95(dd,2H), 2.81–2.65(m,2H), 2.43(s,3H), 2.17(s,3H).

Example 42: N-(3-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)imidazo[1,5-a]pyrazin-6-yl ) Acetamide

Step 1: Preparation of 6-(3-methoxytetrahydrofuran-3-yl)-4-methylpicolinic acid. 2-Bromo-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine (1.00g, 3.67mmol, 1eq.) was dissolved in THF (10mL), and added dropwise at -78°C to n- BuLi (3.67mmol, 1.5mL, 1.0eq.), reacted for 0.5h, then introduced CO ₂ , naturally warmed to room temperature and reacted for 1h, then added 10mL of water to the reaction solution, separated and concentrated to obtain the title compound (0.8g, yield 91%). MS (m/z) = 238.10 [M+H] ⁺ .

Step 2: Preparation of tert-butyl ((5-chloropyrazin-2-yl)methyl)carbamate. 5-chloropyrazine-2-carbaldehyde (1.00 g, 7.02 mmol, 1 eq.), Et ₃ SiH (8.16 g, 70.1 mmol, 10.0 eq.), NH ₂ Boc (1.64 g, 14.03 mmol, 2.0 eq.), TFA (2.40g, 21.0mmol, 3.0eq.) was dissolved in ACN (10mL). After reacting at 25°C for 24h, 10mL of saturated sodium carbonate was added, ACN was spun out, extracted with 10mL of EA, and concentrated to obtain the title compound (1.2g, yield rate of 70%). MS (m/z) = 244.08 [M+H] ⁺ .

Step 3: Preparation of (5-chloropyrazin-2-yl)methanamine. ((5-Chloropyrazin-2-yl)methyl)carbamate tert-butyl ester (1.20 g, 4.92 mmol, 1 eq.) was dissolved in HCl in EA (10 mL), reacted at 25 °C for 16 h, and concentrated to give the title Compound (0.8 g, yield 100%). MS (m/z) = 144.03 [M+H] ⁺ .

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

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

Step 6: N-(3-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)imidazo[1,5-a]pyrazin-6-yl) Preparation of acetamide. 6-Chloro-3-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)imidazo[1,5-a]pyrazine (0.10g, 290μmol, 1eq.), cesium carbonate (283mg, 870μmol, 3.0eq.), acetamide (25.7mg, 435μmol, 1.5eq.), Pd ₂ (dba) ₃ (26.5mg, 29.0μmol, 0.1eq.), X -phos (5.70mg, 58.0μmol, 0.2eq.) was dissolved in 1,4-dioxane (5mL), and reacted at 100°C for 12h under nitrogen protection. The reaction solution was concentrated and purified by column chromatography (PE/EtOAc= 2:1-0:1) to obtain the title compound (0.08g, yield 75%). MS (m/z) = 368.16[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ10.50 (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).

Example 43: N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-methyl-2-oxo-2,3- Dihydro-1H-imidazol[4,5-c]pyridin-6-yl)acetamide

Step 1: Preparation of 6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-amine. 2-Bromo-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine (0.50g, 1.84mmol, 1eq.), dimethylethylenediamine (16.2mg, 183μmol, 0.1eq. ), Cu ₂ O (13.1mg, 91.8μmol, 0.05eq.), potassium carbonate (508mg, 3.67mmol, 2.0eq.) were dissolved in ethylene glycol (5mL) and ammonia water (4.6mL), at 80°C under nitrogen protection Reaction 12h. 20 mL of EA and 20 mL of water were added to the reaction solution, 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] ⁺ .

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

Step 3: Preparation of 6-chloro-N ⁴ -(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)pyridine-3,4-diamine. N-(2-Chloro-5-nitropyridin-4-yl)-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-amine (0.50g, 1.37mmol, 1eq .) and Fe (383mg, 6.85mmol, 5eq.) were dissolved in saturated ammonium chloride solution (10mL) and ethanol (10mL), and reacted at 70°C for 3h. It was spin-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] ⁺ .

Step 4: 6-chloro-1-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1,3-dihydro-2H-imidazo Preparation of [4,5-c]pyridin-2-one. 6-Chloro-N ⁴ -(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)pyridine- 3,4-Diamine (0.40g, 1.19mmol, 1eq.), triphosgene (354mg, 1.19mmol, 1eq.) and TEA (120mg, 1.19mmol, 1eq.) were dissolved in DCM (12mL) and reacted at 20°C 1h. The reaction solution was poured into 10 mL of water, separated and concentrated to obtain the title compound (0.3 g, yield 70%). MS (m/z) = 361.10 [M+H] ⁺ .

Step 5: 6-chloro-1-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-methyl-1,3-dihydro - Preparation of 2H-imidazo[4,5-c]pyridin-2-one. 6-Chloro-1-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine -2-yl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one (0.30g, 831μmol, 1eq.) was dissolved in DMF (2mL), and NaH was added at 20°C (20mg, 831 μ mol, 1eq.), after reacting for 15 minutes, add iodomethane (118mg, 831 μ mol, 1eq.), continue reaction 2h.Reaction solution is poured into 10mL water and adds 10mL EA extraction, separates and concentrates to obtain title compound ( 0.2 g, yield 64%). MS (m/z) = 375.11 [M+H] ⁺ .

Step 6: N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-methyl-2-oxo-2,3-di Preparation of Hydrogen-1H-imidazo[4,5-c]pyridin-6-yl)acetamide. 6-chloro-1-(6-(3-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-methyl-1,3-dihydro-2H- Imidazo[4,5-c]pyridin-2-one (0.20g, 534μmol, 1eq.), acetamide (37.8mg, 640μmol, 1.2eq.), Pd ₂ (dba) ₃ (49mg, 53μmol, 0.1eq .), X-phos (50.87mg, 106μmol, 0.2eq.), cesium carbonate (522mg, 1.60mmol, 3eq.) was dissolved in 1,4-dioxane (5mL), reacted at 100°C under nitrogen protection for 12h Concentration and purification by column chromatography (PE/EtOAc=2:1-0:1) afforded the title compound (0.2 g, yield 94%). MS (m/z)=398.18[M+H] ^{+ .1} H NMR(400MHz,DMSO)δ10.41(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).

Example 44: (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(oxetane-3- ethynyl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

Step 1: (R)-6-chloro-1-(6-(3-(methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2- c] Preparation of pyridine. Dissolve 6-chloro-1H-pyrrolo[3,2-c]pyridine (6.000g, 39.324mmol, 1.00eq.) in 1,4-dioxane (80mL), add (R)-2-Bromo-6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine (10.702g, 39.324mmol, 1.00eq.), K ₂ CO ₃ (16.305g, 0.118mol , 3.0eq.), CuI (0.375mg, 1.966mmol, 0.05eq.), N,N-dimethylethylenediamine (0.348g, 3.933mmol, 0.1eq.), reflux reaction under nitrogen protection for 12h. The reaction The liquid was spin-dried, and the crude product was purified by column chromatography (PE/EtOAc=1:1) to obtain the title compound (11.040 g, yield 81.65%). MS (m/z)=344[M+H] ⁺ .

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

Step 3: (R)-N-(3-iodo-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3, Preparation of 2-c]pyridin-6-yl)acetamide. (R)-N-(1-(6-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c ]pyridin-6-yl)acetamide (9.500g, 25.927mmol, 1.0eq.) was dissolved in DMF (50mL), NIS (8.166g, 36.298mmol, 1.4eq.) was added, and reacted at 60°C for 1h. The reaction solution Cool, pour into water (150mL), extract with EA (200mL), separate liquids, wash the organic phase once with aqueous sodium sulfite solution, separate liquids, wash the organic phase once with saturated NaCl aqueous solution, separate liquids, and wash the organic phase with anhydrous magnesium sulfate Dry, filter, concentrate, and purify by column chromatography (PE/EtOAc=1:1) to give the title compound (6.400 g, yield 35.81%). MS (m/z)=493[M+H] ⁺ .

Step 4: (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(oxetan-3-yl Preparation of ethynyl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide. (R)-N-(3-iodo-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[3,2- c] pyridin-6-yl)acetamide (0.200g, 0.408mmol, 1.0eq.) was dissolved in triethylamine (10mL), and 3-ethynyloxetane (0.0504g, 0.612mmol, 1.5eq. .), Pd(pph ₃ ) ₂ Cl ₂ (0.0288g, 0.0408mmol, 0.1eq.), CuI (0.0308g, 0.1632mmol, 0.4eq.), under nitrogen protection, reacted at 28°C for 15h. The reaction solution was poured into water (20mL), extracted with DCM (50mL), separated, the organic phase was washed once with saturated NH ₄ Cl aqueous solution, separated, the organic phase was washed once with saturated NaCl aqueous solution, separated, and the organic phase was separated. It was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH=30:1) to obtain the title compound (0.080 g, yield 43.90%). MS (m/z) = 447 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.51(s,1H),9.07(s,1H),8.65(d,1H),8.34(s,1H),7.61(s,1H),7.36(s,1H ),4.87(dd,2H),4.70(dd,2H),4.22(m,2H),3.96(m,3H),3.14(s,3H),2.67(m,1H),2.46(m,4H) ,2.11(s,3H).

Example 45: (R)-N-(3-((3-Hydroxyoxetan-3-yl)ethynyl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4 -Methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

The title compound was obtained by referring to the synthetic method of Example 44. MS (m/z) = 463 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.54(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).

Example 46: (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-((3-methyloxa Cyclobut-3-yl)ethynyl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

The title compound was obtained by referring to the synthetic method of Example 44. MS (m/z) = 461 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.52(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).

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

Step 1: (R)-3-((6-Acetamido-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-1H-pyrrolo[ Preparation of tert-butyl 3,2-c]pyridin-3-yl)ethynyl)azetidine-1-carboxylate. The title compound was obtained by referring to the synthetic method of Example 44. MS (m/z) = 546 [M+H] ⁺ .

Step 2: (R)-N-(3-(azetidin-3-ylethynyl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridine- Preparation of 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-carboxylate tert-butyl ester (0.15 g, 0.275 mmol, 1.0 eq.) was dissolved in DCM (10 mL) and TFA was added at room temperature ( 4mL), stirred for 30min. Pour the reaction system into 30 mL of water, adjust the pH to 10 with sodium carbonate, extract with 100 mL of DCM, separate the liquids, wash the organic phase once with saturated NH ₄ Cl aqueous solution, separate the liquids, wash the organic phase once with saturated NaCl aqueous solution, and separate the liquids , the organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated to give the title compound (0.12 g, yield 95%). MS (m/z) = 446 [M+H] ⁺ .

Step 3: (R)-N-(3-((1-acetylazetidin-3-yl)ethynyl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4 - Preparation of -methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide. (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 (0.12g, 0.269mmol, 1.0eq.) was dissolved in DCM (10mL), and acetic anhydride (0.042g, 0.405mmol, 1.5eq.) stirred for 30min. Pour the system into 10 mL of water, adjust the pH to 10 with saturated sodium carbonate aqueous solution, extract with 50 mL of DCM, separate the liquids, wash the organic phase once with a saturated NH ₄ Cl aqueous solution, separate the liquids, wash the organic phase once with a saturated NaCl aqueous solution, The layers were separated, and 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 (0.045 g, yield 34.4%). MS (m/z) = 488 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.52(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).

Example 48: (R)-N-(3-((3-methoxyoxetan-3-yl)ethynyl)-1-(6-(3-methoxytetrahydrofuran-3-yl) -4-methylpyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

The title compound was obtained by referring to the synthetic method of Example 44. MS (m/z) = 477 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.54(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).

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

The title compound was obtained by referring to the synthetic method of Example 44. MS (m/z) = 435 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.50(s,1H),9.07(s,1H),8.60(d,1H),8.27(s,1H),7.60(s,1H),7.34(s,1H ),4.95(t,1H),4.21(d,1H),4.01(m,3H),3.65(q,2H),3.15(s,3H),2.65(m,3H),2.46(m,4H) ,2.11(s,3H).

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

(R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(1-methylazetidine- 3-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide (0.10g, 0.229mmol, 1.0eq.) was dissolved in methanol (10mL), and 4M aqueous sodium hydroxide solution was added (5mL), heated to 60°C, reacted for 15h. The system was spin-dried, and purified by column chromatography (DCM/MeOH=30:1) to obtain the title compound. MS (m/z) = 394 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ8.33(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) .

Example 51: (R)-N-(3-(1-Acetazetidin-3-yl)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methyl Pyridin-2-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

The title compound was obtained by referring to the synthetic method of Example 47. MS (m/z) = 464 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.42(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).

Example 52 was prepared in a similar manner with reference to Example 51.

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

The title compound was obtained by referring to the synthetic method of Example 35. MS (m/z) = 464 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.41(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).

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

(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 (0.125g, 0.291mmol, 1.0eq.) was dissolved in THF (10mL), and 2,2,2-trifluoroethyl Trifluoromethanesulfonate (0.081g, 0.349mmol, 1.2eq.), triethylamine (0.0589g, 0.582mmol, 2.0eq.), react at room temperature for 15h. Pour the system into 20mL of water, extract with _80mL of ethyl acetate, and separate the liquids. It was dried over 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 (400MHz,DMSO)δ10.39(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).

Example 55 was prepared in a similar manner with reference to Example 54.

Example 56: N-(3-(2-Methyl-2-azaspiro[3.3]heptyl-6-yl)-1-(4-methylpyridin-2-yl)-1H-pyrrolo[ 3,2-c]pyridin-6-yl)acetamide

The title compound was obtained by referring to the synthetic methods of Examples 1 and 35. MS (m/z) = 376 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.38(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) .

Example 57: (R)-1-(1-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(1-methyl Azetidin-3-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)urea

(R)-1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(1-methylazetidin-3-yl) -1H-pyrrolo[3,2-c]pyridin-6-amine (0.20g, 0.508mmol, 1.0eq.) was dissolved in acetic acid (10mL) and water (30mL), heated to 90°C, and divided within one hour Potassium cyanate (2.06g, 25.41mmol, 50eq.) was added in batches and reacted for 20min. Cool the reaction solution to room temperature, adjust the pH to about 9 with potassium carbonate, extract with dichloromethane, combine and concentrate the organic phases, and purify by column chromatography (DCM/MeOH=30:1) to obtain the title compound (0.01 g, yield 4.5 %). MS (m/z) = 437 [M+H] ⁺ . ¹ H NMR (400MHz, CDCl ₃ )δ9.71(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).

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

The title compound was obtained by referring to the synthetic method of Examples 50 and 57. MS (m/z) = 477 [M+H] ⁺ . ¹ H NMR (400MHz,MeOD)δ8.48(s,1H),8.16(s,1H),7.65(d,1H),7.41(s,1H),7.33(s,1H),4.25(d,1H ),4.14(m,4H),4.00(s,2H),3.79(s,2H),3.67(m,1H),3.25(s,3H),2.81(m,2H),2.70(m,4H) ,2.54(m,6H).

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

The title compound was obtained by referring to the synthetic method of Examples 1 and 44. MS (m/z) = 433 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ12.89(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).

Examples 60-62 were prepared in a similar manner with reference to Example 35.

The compounds of Examples 63-64 were prepared by referring to the method of Example 1.

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

The title compound was obtained by referring to the synthetic methods of Examples 1 and 35. MS (m/z) = 462 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.73(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).

Example 66 4-(6-Acetamido-3-(2-methyl-2-azaspiro[3.3]heptane-6-yl)-1H-pyrrolo[3,2-c]pyridine-1- base)-2,6-dimethylbenzoic acid methyl ester

Referring to the synthesis method of Example 56, the title compound was obtained. MS (m/z) = 447 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.44(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).

Example 67 (R)-N-(3-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)imidazo[1,5-a]pyrazine- 6-yl)acetamide

The title compound was prepared by referring to the synthetic method of Example 42. MS (m/z) = 368.42 [M+H] ⁺ . ¹ H NMR (400MHz,DMSO)δ10.51(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).

The compounds of Examples 68-69 were prepared by referring to the method of Example 1.

Example 70 (R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(2-(methyl-d3) -2-Azaspiro[3.3]heptane-6-yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide

(R)-N-(1-(6-(3-methoxytetrahydrofuran-3-yl)-4-methylpyridin-2-yl)-3-(2-azaspiro[3.3]hept-6 -yl)-1H-pyrrolo[3,2-c]pyridin-6-yl)acetamide (0.100 g, 216.659 μmol, 1 eq) was dissolved in DMF (10 mL), and NaHCO ₃ (36.402 mg, 433.318 μmol, 2 eq .), deuteroiodomethane (21.984mg, 151.661μmol, 0.7eq.), react at room temperature for 15h. The reaction solution was poured into 50 mL of water, extracted with 100 mL of EtOAC, separated, the organic phase was washed once with saturated NaCl aqueous solution, separated, the organic phase was dried with anhydrous magnesium sulfate, filtered, and concentrated. Using DCM:MeOH=20:1 to climb the plate, the title compound (0.002g) was obtained. MS (m/z) = 479.61 [M+H] ⁺ . ¹ H NMR (400MHz,MeOD)δ8.98(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).

Examples 71-72 were prepared in a similar manner to Example 1 and Example 56.

Example 73a and Example 73b were prepared in a similar manner with reference to Example 28a and Example 28b.

biological evaluation

Experimental example 1 TYK2 JH2, TYK2 JH1, JAK1, JAK2, JAK3 kinase activity inhibition assay

TYK2 JH2 Kinase Activity Inhibition Assay

TYK2 JH2 (N-His-Tev, 575-869) was expressed in Sf9, the protein expression was from our laboratory, the fluorescein-labeled probe was synthesized by our company, and Anti-6xHis-terbium labeled antibody was purchased from Cisbio. High purity Hepes, NaCl, _MgCl2 , DTT, BSA, Tween-20 and DMSO were purchased from Sigma.

The assay buffer used in the experiment consisted of 20mM Hepes pH 7.5, 150mM NaCl, 10mM MgCl ₂ , 2mM DTT, 50μg/mL BSA, and 0.015% Tween-20. Prepare the stock solution of the compound to be tested in DMSO, and perform a three-fold concentration gradient dilution with DMSO for 12 points according to the experimental requirements. 4% DMSO compound, TYK2 JH2 enzyme, fluorescein-labeled probe, Anti-6xHis-terbium labeled antibody were prepared using 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 to OptiPlate-384 White Opaque plates, covered with membrane, 800 rpm for 1 min, and incubated at room temperature for 1.5 h. The final concentrations of compound, TYK2 JH2 enzyme, luciferin-labeled probe and Anti-6xHis-terbium labeled antibody in 4% DMSO were 1%, 2.5nM, 50nM and 1x, respectively. After 1.5 h, the plate was read on a SPARK multi-template reader of TECAN (Switzerland), the excitation light wavelength was 340 nm, and the emission light wavelengths were 520 nm and 485 nm, respectively. _IC50 values of inhibitors were obtained by using Prism 8 (La Jolla, CA).

TYK2 JH1, JAK1, JAK2, JAK3 Kinase Activity Inhibition Assay

TYK2 JH1 (NP_003322.3), JAK2 (NP_004963.1) and JA3 (NP_000206.2) were purchased from carna company, JAK1 (N-GST-his-TEV, 850-1154) was expressed in HI5, from our laboratory For protein expression, TK kit was purchased from Cisbio. High purity ATP, _MgCl2 , _MnCl2 , DTT and DMSO were purchased from Sigma.

The assay buffer used in the TYK2 JH1 experiment consisted of 5mM MgCl ₂ , 1mM MnCl ₂ , 1mM DTT, 12.5μM SEB and 1x Enzymatic buffer.

The assay buffer used in the JAK2/3 experiment consisted of 5mM MgCl ₂ , 1mM DTT and 1x Enzymatic buffer.

The assay buffer used in the JAK1 experiment consisted of 5mM MgCl ₂ , 1mM MnCl ₂ , 1mM DTT and 1x Enzymatic buffer.

Prepare the stock solution of the compound to be tested in DMSO, and perform a three-fold concentration gradient dilution with DMSO for 12 points according to the experimental requirements. 4% DMSO compound, enzyme, TK-Substrate and ATP were prepared using assay buffer. After the preparation was completed, 2.5 μL 4% DMSO compound, 2.5 μL enzyme, and 5 μL TK-Substrate/ATP mixture were added to the OptiPlate-384 White Opaque plate In the medium, cover the membrane, 800 rpm for 1min, and incubate at room temperature for 1h. The final concentration of the compound is 1% in 4% DMSO, the final concentration of TYK2 JH1, JAK1, JAK2 and JAK3 enzyme is 0.2ng/μL, 10ng/μL, 0.125ng/μL and 0.3ng/μL, and the final concentration of TK-Substrate/ATP are 0.25 μM and 3 μM, respectively. After 1 h, 5 μL each of TK-Antibody-Cryptate antibody and Streptavidin-XL665 diluted in HTRF detection buffer were added, and incubated at room temperature for 1 h. Final concentrations were 1x and 15.61 nM. After 1 h, the plate was read on the SPARK multi-template reader of TECAN (Switzerland), the excitation light wavelength was 320 nm, and the emission light wavelengths were 665 nm and 620 nm, respectively. _IC50 values of inhibitors were obtained by using Prism 8 (La Jolla, CA).

Table 1 shows the IC ₅₀ values of the representative compounds of the present application for TYK2 JH2, TYK2 JH1, JAK1, JAK2, and JAK3 kinase activity inhibition assays.

Table 1

Examples TYK2 JH2 TYK2 JH1 JAK1 (nM) JAK2(nM) JAK3(nM)

Number (nM) (nM) 1 2.2 2 13.0 3 18.7 4 239.6 5 114.7 6 290.4 7 31.2 8 2.4 9 5 10 2.9 11 5.1 12 1.0 >10000 2480 13 >1000 14 >100 15 0.6 >10000 >10000 16 3.1 17 1.9 >10000 8488 18 5.7 19 >1000 20 3.9 twenty one 1.6 twenty two 1.8 twenty three 1.8 twenty four 3.3 >10000 >10000 >10000 >10000 25 0.4 26 5.3 27 2.1 >10000 >10000 >10000 >10000 28a 1 28b 1.4 29a 1.7 >10000 >10000 >10000 >10000 29b 1.6 >10000 >10000 >10000 30 3.8

31 4.1 32 4.6 33 3.9 34a 2.2 34b 2.6 35 2.7 >10000 >10000 >10000 398.3 36a 3.8 36b 2.5 >10000 37a 2 37b 4.3 >10000 38 2 39 2.7 6159 40 0.5 41a 40.4 41b 31.9 42 20.2 43 147.4 44 6.9 45 12.9 46 15.3 47 3.6 48 27.5 49 16.1 50 186 51 2.3 52 1.5 53 4.9 54 3.1 55 1.9 56 11.2 57 2.2 58 1.8 59 2.9

60 2.6 61 3.6 62 9.8 63 64 65 203 66 1.3 67 17 68 69 6.4 70 71 72 18 73a 3.4 73b 2

Note: Blank means untested.

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

Experimental example 2 Permeability determination of Caco-2 cell monolayer

Material:

Caco-2 cells and experimental reagent solvents were obtained from commercial sources

Reference Compound 1 and Reference Compound 2 were prepared by referring to the method in International Patent Publication WO2019178079A1.

General method for the permeability assay of Caco-2 cell monolayers:

1. Preheating: Preheat the HBSS buffer in a 37°C water bath.

2. Take out the sample from -20°C, and ultrasonically treat it for not less than 1 minute.

3. Buffer preparation

Administration side blank solution:

A-to-B direction:

HBSS+ with 0.3% DMSO and 5 μM Lucifer Yellow: Add 150 μL of DMSO and 125 μL of 2 mM Lucifer Yellow to 50 mL of HBSS+ buffer (pH 7.4).

HBSS+ with 0.1% DMSO and 5 μM Lucifer Yellow: Add 50 μL of DMSO and 125 μL of 2 mM Lucifer Yellow to 50 mL of HBSS+ buffer (pH 7.4).

B-to-A direction:

HBSS+ with 0.3% DMSO: Add 150 μL DMSO to 50 mL of HBSS+ buffer (pH 7.4).

HBSS+ with 0.1% DMSO: Add 50 μL DMSO to 50 mL of HBSS+ buffer (pH 7.4).

Receiving side solution:

A-to-B direction:

HBSS+ with 0.4% DMSO: Add 200 μL DMSO to 50 mL of HBSS+ buffer (pH 7.4).

B-to-A direction:

HBSS+ with 0.4% DMSO and 5 μM Lucifer Yellow: Add 200 μL of DMSO and 50 μL of 5 mM Lucifer Yellow to 50 mL of HBSS+ buffer (pH 7.4).

4. Measure the transmembrane resistance:

Take out the cell culture plate from the incubator, rinse the cells twice with HBSS buffer (add 400 μL to each well of the upper cell plate, add 25 mL to the lower support plate), and measure the transmembrane resistance with a Millicell ERS resistance meter at room temperature.

5. Place the administration side solution in a centrifuge and centrifuge at 4000rpm for 5 minutes. The supernatant was collected as the administration-side solution.

6. Administration:

A-B (administration side): 600 μL of A-to-B administration solution (100 μL for sample collection at the beginning of culture of Luciferus yellow, 100 μL for sample collection at the beginning of culture).

AB (receiving side): 800 μL of HBSS ⁺ with 0.4% DMSO.

B-A (dosing side): 900 μL of B-to-A dosing solution (100 μL for sample collection at the beginning of culture).

BA (receiving side): 500 μL of HBSS ⁺ 0.4% DMSO with 5 μM Luciferin (100 μL for sample collection at the beginning of the Luciferin culture).

7. Put the administered upper plate and lower plate into the 37°C incubator to preheat for 5 minutes. Then take 100 μL from the administration side for sample collection at the beginning (A-B D0, B-A D0); take 100 μL from each well of the upper plate and place it in a 96-well black fluorescence measurement plate for sample collection at the beginning of Lucifer yellow culture (D0) .

8. Put the upper plate and the lower plate together to start the permeability test, and incubate at 37°C for 90 minutes.

9. After the incubation, separate the upper plate from the lower plate, take out 100 μL sample from the administration side and put it into a 96-well black fluorescence measurement plate, measure the amount of Lucifer yellow after 90 minutes (excitation wavelength 485nm, emission wavelength 535nm) .

10. The samples of the administration solution and the receiving solution were diluted with 0.4% DMSO HBSS buffer solution, then mixed with CAN containing the internal standard and sent to liquid chromatography-mass spectrometry analysis.

11. Data processing

Transmembrane resistance (TEER) = (resistance value sample – resistance value blank) × membrane area

Lucifer Yellow Permeability:

Apparent permeability coefficient (P _app ) = (volume of receiving side/(membrane area × culture time))×(fluorescence value of receiving side at the end of culture – fluorescence value of blank solution)/((fluorescence value of administration side at the beginning of culture – Fluorescence value of blank solution) × dilution factor)

Test compound permeability:

Apparent permeability coefficient (P _app )=(receiving side volume/(membrane area×culture time))×(drug concentration on the receiving side at the end of culture/(drug concentration on the administration side at the beginning of culture×dilution factor)

Millipore cell culture plate (PSHT 010 R5) was used in this experiment: membrane area = 0.7cm ² , receiving side volume = 0.8mL (A-to-B) or 0.4mL (B-to-A), incubation time = 90 minutes.

Compound Recovery:

% recovery rate=100×(the total amount of compounds on the administration side in 90 minutes+the total amount of compounds on the receiving side in 90 minutes)/(the total amount of compounds on the administration side in 0 minutes).

See Table 2 for the apparent permeability coefficients of representative compounds of the present application and reference compound 1 and reference compound 2.

Table 2

The results show that the ratio of the apparent permeability coefficient B-A/A-B of the compound of the application is lower than that of the control compound, the absorption capacity of the drug is better than that of the control compound, and it is not easy to efflux, and it is easier to produce drug effect in the cell.

Claims (16)

1. A compound of formula (I):

   

   or a tautomer, meso, racemate, enantiomer, diastereomer or a pharmaceutically acceptable salt thereof,

   wherein A is ₁ 、A ₂ 、A ₄ 、A ₅ 、A ₆ 、A ₇ 、A ₈ Selected from C or N, A ₃ Selected from C, N or

   

   And when A ₂ When N is N, A is ₄ 、A ₅ 、A ₆ 、A ₇ 、A ₈ At least 1 of them is N;

   

   is a saturated or unsaturated ring;

   R ₁ Selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms being optionally substituted with 1-3R _a Substitution;

   R ₂ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms being optionally substituted with 1-3R _a Substitution;

   R ₄ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms being optionally substituted with 1-3R _a Substitution;

   each R _a 、R _b Are independently selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, C ₃ -C ₆ Cycloalkyl, C ₃ -C ₆ Heterocycloalkyl, halogen, hydroxy, cyano, nitro, -C (O) NR _c R _d 、-C(O)R _c 、-(CH ₂ ) _n C(O)OR _c 、-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) ₂ R _c Said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl being optionally substituted with 1-3R _c Substitution;

   each R _c 、R _d Independently selected from hydrogen, halogen, carbonyl, -C (O) CH ₃ Hydroxyl, cyano, nitro, C ₁ -C ₆ Alkyl, halogenated C ₁ -C ₆ Alkyl, C ₃ -C ₆ Cycloalkyl or halo C ₃ -C ₆ Cycloalkyl; and is also provided with

   Each n is independently selected from 0, 1, 2 or 3.
2. The compound represented by the general formula (I) according to claim 1, which is a compound represented by the general formula (Ia):

   

   or a tautomer, meso, racemate, enantiomer, diastereomer or a pharmaceutically acceptable salt thereof,

   wherein A is ₁ 、A ₂ 、A ₄ 、A ₅ 、A ₆ 、A ₇ 、A ₈ Selected from C or N, A ₃ Selected from C, N or

   

   And when A ₂ When N is N, A is ₄ 、A ₅ 、A ₆ 、A ₇ 、A ₈ At least 1 of them is N;

   

   is a saturated or unsaturated ring;

   R ₁ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms being optionally substituted with 1-3R _a Substitution;

   R ₂ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms being optionally substituted with 1-3R _a Substitution;

   R ₃ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms being optionally substituted with 1-3R _a Substitution;

   R ₄ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or containing 0-3 hetero atomsThe 3-10 membered saturated or unsaturated ring of said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms optionally substituted with 1-3R _a Substitution;

   each R _a 、R _b Are independently selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, C ₃ -C ₆ Cycloalkyl, C ₃ -C ₆ Heterocycloalkyl, halogen, hydroxy, cyano, nitro, -C (O) NR _c R _d 、-C(O)R _c 、-(CH ₂ ) _n C(O)OR _c 、-OR _c 、-(CH ₂ ) _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) ₂ R _c Said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl being optionally substituted with 1-3R _c Substitution;

   each R _c 、R _d Independently selected from hydrogen, halogen, carbonyl, -C (O) CH ₃ Hydroxyl, cyano, nitro, C ₁ -C ₆ Alkyl, halogenated C ₁ -C ₆ Alkyl, C ₃ -C ₆ Cycloalkyl or halo C ₃ -C ₆ Cycloalkyl; and is also provided with

   Each n is independently selected from 0, 1, 2 or 3.
3. The compound represented by the general formula (I) according to claim 1, which is a compound represented by the general formula (Ib):

   

   or a tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt thereof, wherein A ₁ 、A ₃ Selected from C or N;

   R ₁ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms being optionally substituted with 1-3R _a Substitution;

   R ₂ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 hetero atoms being optionally substituted with 1-3R _a Substitution;

   R ₃ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, halo C ₁ -C ₆ Alkyl, halogenated C ₂ -C ₆ Alkenyl, halo C ₂ -C ₆ Alkynyl, halogen, cyano, nitro, -C (O) NR _a R _b 、-C(O)R _a 、-C(O)OR _a 、-OR _a 、-(CH ₂ ) _n OR _a 、-NR _a R _b or-S (O) ₂ R _a ；

   R ₄ ' selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, halo C ₁ -C ₆ Alkyl, halogenated C ₂ -C ₆ Alkenyl, halo C ₂ -C ₆ Alkynyl, C ₃ -C ₆ Cycloalkyl, halo C ₃ -C ₆ Cycloalkyl, halogen, cyano, nitro or-NR _a R _b ；

   Each R _a 、R _b Are independently selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, C ₃ -C ₆ Cycloalkyl, C ₃ -C ₆ Heterocycloalkyl, halogen, hydroxy, cyano, nitro, -C (O) NR _c R _d 、-C(O)R _c 、-(CH ₂ ) _n C(O)OR _c 、-OR _c 、-(CH ₂ ) _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) ₂ R _c Said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl being optionally substituted with 1-3R _c Substitution;

   each R _c 、R _d Independently selected from hydrogen, halogen, carbonyl, -C (O) CH ₃ Hydroxyl, cyano, nitro, C ₁ -C ₆ Alkyl, halogenated C ₁ -C ₆ Alkyl, C ₃ -C ₆ Cycloalkyl or halo C ₃ -C ₆ Cycloalkyl; and is also provided with

   Each n is independently selected from 0, 1, 2 or 3.
4. The compound represented by the general formula (I) according to claim 1, which is a compound represented by the general formula (Ic):

   

   or a tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt thereof, wherein A ₃ Selected from C or N;

   R ₁ selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ 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) ₂ R _a Or contains 0-3 hetero-sources3-10 membered saturated or unsaturated ring of the sub-group, said alkyl, alkenyl, alkynyl, 3-10 membered saturated or unsaturated ring containing 0-3 heteroatoms optionally substituted with 1-3R _a Substitution;

   R ₂ selected from hydrogen, C ₁ -C ₆ Alkyl, halogenated C ₁ -C ₆ Alkyl, halogen, cyano, -C (O) OR _a 、-(CH ₂ ) _n OR _a or-NH ₂ ；

   R ₃ Selected from hydrogen, C ₁ -C ₆ Alkyl OR-OR _a ；

   R ₄ ' selected from hydrogen, C ₁ -C ₆ Alkyl, halogenated C ₁ -C ₆ Alkyl, C ₃ -C ₆ Cycloalkyl, halogen or-NH ₂ ；

   Each R _a 、R _b Are independently selected from hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, C ₃ -C ₆ Cycloalkyl, C ₃ -C ₆ Heterocycloalkyl, halogen, hydroxy, cyano, nitro, -C (O) NR _c R _d 、-C(O)R _c 、-(CH ₂ ) _n C(O)OR _c 、-OR _c 、-(CH ₂ ) _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) ₂ R _c Said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl being optionally substituted with 1-3R _c Substitution;

   each R _c 、R _d Independently selected from hydrogen, halogen, carbonyl, -C (O) CH ₃ Hydroxyl, cyano, nitro, C ₁ -C ₆ Alkyl, halogenated C ₁ -C ₆ Alkyl, C ₃ -C ₆ Cycloalkyl or halo C ₃ -C ₆ Cycloalkyl; and is also provided with

   Each n is independently selected from 0, 1, 2 or 3.
5. The compound of any one of claims 1-4, wherein R ₁ Selected from hydrogen, -CH ₃ 、

   

   

   

   -CN、

   

   -F、

   

   
6. The compound of any one of claims 1-4, wherein R ₂ Selected from hydrogen, -CH ₃ 、

   

   
7. The compound of any one of claims 1-4, wherein R ₃ Is that

   
8. The compound according to claim 1 or 2, wherein R ₄ Is hydrogen, -C (O) CH ₃ 、

   

   or-C (O) NH ₂ 。
9. A compound of formula (I) according to claim 1, selected from:

   

   

   

   

   
10. use of a compound according to any one of claims 1-9, an isomer thereof or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for diseases mediated by TYK 2.
11. The use of claim 10, wherein the TYK 2-mediated disease is an autoimmune disease, an inflammatory disease, a proliferative disease, an endocrine disease, a neurological disease, or a disease associated with transplantation.
12. The use according to claim 11, wherein the disease is an autoimmune disease.
13. The use of claim 12, wherein the autoimmune disease is selected from type 1 diabetes, ankylosing spondylitis, systemic lupus erythematosus, multiple sclerosis, systemic sclerosis, psoriasis, crohn's disease, ulcerative colitis, or inflammatory bowel disease.
14. The use according to claim 11, wherein the disease is an inflammatory disease.
15. The use according to claim 14, wherein the inflammatory disease is selected from the group consisting of rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, psoriasis, crohn's disease, ulcerative colitis, and inflammatory bowel disease.
16. A pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of claims 1-9, an isomer thereof, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient.