Compound with kinase inhibition function as well as preparation and application thereof

Abstract

The invention provides an HPK1 kinase inhibitor as well as preparation and application thereof. Specifically, the invention provides a compound as shown in formula (I), and the definition of each group is described in the specification. The compound has excellent HPK1 inhibitory activity, so that the compound can be used for preparing pharmaceutical compositions for treating cancers and other diseases related to HPK1 activity. # imgabs0 #

Description

A class of compounds with kinase inhibitory function and their preparation and application Technical Field

[0001] The present invention relates to the field of small molecule drugs, and in particular, to a kinase inhibitor and its preparation and use. Background Art

[0002] T cells, B cells, and dendritic cells (DCs) are powerful weapons of the human immune system against foreign invaders such as viruses and bacteria, as well as infected or abnormal cells such as cancerous cells. The human body possesses a complex and sophisticated regulatory system to ensure the proper functioning of the immune system. When cancerous cells appear in the body, if the immune system is unable to completely eliminate them, these immune-evading cancer cells proliferate abnormally and form tumors. Traditional cancer treatments primarily rely on surgery, radiotherapy, chemotherapy, and molecularly targeted drugs. However, surgical resection is often not a viable option for many forms of tumors or cancers. While radiotherapy and chemotherapy target tumor cells, they also damage some healthy cells. Tumor cells are prone to mutation and can develop resistance to drugs specifically targeting tumor cells, making cancer treatment challenging. Harnessing the patient's own immune system to overcome the immune evasion strategies employed by tumor cells and enhance anti-tumor immunity has become a new cancer treatment strategy. One such strategy overcomes immune evasion by suppressing negative regulators of the immune response that normally maintain peripheral tolerance, allowing tumor antigens to be recognized as non-self antigens. Hematopoietic progenitor cell kinase 1 (HPK1), also known as MAP4K1 (a member of the MAP4K family), is a negative regulator of activation responses in dendritic cells (DCs), T cells, and B cells. Inhibiting its activity can specifically enhance anti-tumor immunity. HPK1 is primarily expressed by hematopoietic cells, including early hematopoietic progenitor cells. In T cells, HPK1 is believed to negatively regulate T cell activation by phosphorylating the downstream SLP76 protein at Ser376 and the Gads protein at Thr254 and recruiting 14-3-3 proteins for degradation, thereby reducing the persistence of signaling microclusters. HPK1 is also activated in response to prostaglandins (PGE2), which are commonly secreted by tumors, helping tumor cells evade the immune system. HPK1 also inhibits AP-1, which plays a role in promoting cell proliferation, inhibiting differentiation, and promoting tumor cell invasion and metastasis during tumor formation and progression. Targeted disruption of HPK1 kinase alleles can enhance T cell production of Th1 cytokines (IL-2, IFNγ, etc.) in TCR responses. HPK1 plays multiple roles in immunity and is implicated in the pathogenesis of autoimmune diseases, cancer, and inflammatory responses. HPK1 kinase knockout (HPK1- / -) T cells proliferate much faster than wild-type monoclonal antibodies, and mice transfected with HPK1- / - T cells can resist tumor growth. Dendritic cells (DCs) lacking HPK1 kinase have better antigen presentation capacity than wild-type cells and can better express anti-tumor immune responses. Furthermore, animal studies have demonstrated that HPK1 inhibition and PD-1 / PD-L1 antibody drugs have significant synergistic anti-tumor activity.Therefore, HPK1 kinase plays a key role in disease treatment, especially cancer treatment.

[0003] Currently, there are no drugs targeting this target on the market. In order to meet the huge clinical demand in the future, this field needs to design and develop selective and highly active small molecule HPK1 inhibitors to provide new oral drugs for the treatment of immune-related diseases, especially tumors, which can be used alone or in combination with chemotherapy, radiotherapy, tumor-targeted drugs, other tumor immunotherapeutics (small molecule compounds and antibodies) and tumor vaccines.

[0004] Summary of the Invention

[0005] The purpose of the present invention is to provide a small molecule HPK1 inhibitor with selectivity and high activity.

[0006] The first aspect of the present invention provides a compound represented by the following formula I or formula II, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof;

[0007] X 1 Selected from the following group: CR 3 NR 3 ;

[0008] X 2 Selected from the group consisting of: N, CH;

[0009] X 4 N or CH; Z 1 is N or C; and It is an aromatic ring;

[0010] R 3 Selected from the following group: H, C 1-6 Alkyl, C 3-8 Cycloalkyl (preferably C 3-6 cycloalkyl), a 4-8 membered heterocyclic group containing 1-3 heteroatoms selected from N, O, and S, a 4-8 membered (preferably 5 membered) heteroaryl group containing 1-3 heteroatoms selected from N, O, and S, and the above groups may be further substituted by 1-3 groups selected from the following groups: D, -NR 11 R 12 、-CN、halogen、C 1-6 Alkyl, C 1-6 Alkoxy, -C(O)R 11 、-SO2R 18 ;

[0011] R 11 , R 12 Selected from H, C 1-3 Alkyl, or R 11 、R 12 Together with the atoms to which they are attached, they form a 4-6 membered heterocyclic ring;

[0012] R 18 C 1-3 alkyl;

[0013] R 1 for: Among them, R a and R b Each independently selected from the group consisting of: H, C 1-6 Alkyl, phenyl, 5-7 membered heteroaryl, C 3-8 Cycloalkyl, C 1-6 Alkenyl, benzyl, C 5-8 Spiroalkyl and C 5-8 Bridged cycloalkyl, 4-12 membered (preferably 4-8 membered) heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O, 5-7 membered heteroaromatic substituted C 1-6 Alkyl, or R a and R b The atoms connected to it together form C 3-8 Cycloalkyl;

[0014] And the R a or R b , or R a and R b The ring formed by the atoms connected thereto may be further substituted by 1-3 groups selected from the group consisting of halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl;

[0015] R c Selected from the group consisting of: -NR 16 R 17 ,-OH;R 16 , R 17 Each independently selected from H, C 1-3 alkyl;

[0016] R 2 It has the structure shown below:

[0017] Where n is 0 or 1;

[0018] Y 1 N or CH;

[0019] Each Y 4 and Y 5 Each independently selected from the following group: O, NR 15 、C(R 13 R 14 ); each R 13 , R14 , R 15 Each independently selected from the following group: H, C 1-6 alkyl;

[0020] R 4 、R 5 Each independently selected from the group consisting of: H, C 1-6 Alkyl (preferably C 1-3 alkyl);

[0021] In the compound of formula I, each chiral center is in R configuration or S configuration.

[0022] In another preferred embodiment, the compound of formula I has the structure shown in the following formula I-1 or I-2:

[0023] Among them, X 1 Selected from the following group: NR 3 , CR 3 ;

[0024] X 2 Selected from the group consisting of: N, CH;

[0025] X 4 N or CH;

[0026] R 1 、R 2 The definition is as stated in claim 1.

[0027] In another preferred embodiment, the R a and R b Each independently selected from the group consisting of: H, C 1-6 Alkyl, phenyl, 5-7 membered heteroaryl, C 3-8 Cycloalkyl, C 1-6 alkenyl, benzyl, a 4-8 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O; or R a and R b The atoms connected to it together form C 3-8 Cycloalkyl; and said R a or R b It may be further substituted by 1-3 groups selected from the group consisting of halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl.

[0028] In another preferred embodiment, the R a and R b Each independently selected from the following group: C 1-4 Alkyl, C 3-6Cycloalkyl, a 4-6 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O; or R a and R b The atoms connected to it together form C 3-8 Cycloalkyl, or a 4-6 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O; and said R a or R b , or R a and R b The group formed by the atoms connected thereto may be further substituted by 1-3 substituents selected from the group consisting of halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl.

[0029] In another preferred embodiment, R 3 Select from the following group: C 1-6 Alkyl, C 3-8 Cycloalkyl (preferably C 3-6 cycloalkyl), a 4-8 membered heterocyclic group containing 1-3 heteroatoms selected from N, O, and S, a 4-8 membered (preferably 5 membered) heteroaryl group containing 1-3 heteroatoms selected from N, O, and S, and the above groups may be further substituted by 1-3 groups selected from the following groups: D, -NR 11 R 12 , halogen, C 1-6 Alkyl, C 1-6 alkoxy;

[0030] R 11 , R 12 Selected from H, C 1-3 Alkyl, or R 11 、R 12 Together with the atoms they are connected to, they form a 4-6 membered heterocyclic ring.

[0031] In another preferred embodiment, the compound of formula I has a structure selected from the following group:

[0032] In another preferred embodiment, the R2 is selected from the following group:

[0033] In another preferred embodiment, the R 2 Select from the following groups:

[0034] In another preferred embodiment, the compound of formula I is selected from the following group:

[0035] In another preferred embodiment, the compound of formula I is selected from the following group:

[0036] In another preferred embodiment, the compound of formula I is selected from the following group:

[0037] In another preferred embodiment, the compound of formula I is selected from the following group:

[0038] The second aspect of the present invention provides a pharmaceutical composition comprising (1) a compound as described in the first aspect of the present invention or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt, hydrate or solvate thereof; and (2) a pharmaceutically acceptable carrier.

[0039] The third aspect of the present invention provides a use of the compound according to the first aspect of the present invention or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition according to the second aspect of the present invention, characterized in that it is used to prepare a pharmaceutical composition for preventing and / or treating diseases related to the activity or expression of HPK1 kinase.

[0040] In another preferred embodiment, the disease is selected from the group consisting of lymphoma, blastoma, medulloblastoma, retinoblastoma, sarcoma, liposarcoma, synovial cell sarcoma, neuroendocrine tumor, carcinoid tumor, gastrinoma, islet cell carcinoma, mesothelioma, schwannoma, acoustic neuroma, meningioma, adenocarcinoma, melanoma, leukemia or lymphoid malignancy, squamous cell carcinoma, epithelial squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, adenocarcinoma lung cancer, squamous cell lung cancer, peritoneal cancer, hepatocellular carcinoma, gastric cancer, intestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, breast cancer, metastasis. Metastatic breast cancer, colon cancer, rectal cancer, colorectal cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, anal cancer, penile cancer, Merkel cell carcinoma, esophageal cancer, biliary tract cancer, head and neck cancer, hematological malignancies, nasopharyngeal cancer, multiple myeloma, Ohba villous adenoma, non-Hodgkin's lymphoma, bone cancer, testicular cancer, Hodgkin's disease, spermatoma, oral cancer, brain cancer, skin cancer, breast duct cancer, renal pelvis cancer, Wilms' tumor, esophageal adenoma, retinoblastoma, glioma, neurofibroma, gastrointestinal stromal tumor, carcinoma in situ, endometrial cancer and myelodysplastic syndrome, etc.

[0041] In a fourth aspect, the present invention provides a combined tumor chemotherapy regimen of the compound according to the first aspect of the present invention and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates and deuterated compounds and other tumor immunotherapeutic agents, wherein the other tumor immunotherapeutic agents are selected from the following groups: small molecule compounds and antibodies (including but not limited to PD-1, PD-L1, CTLA-4, TIM-3, TGF-β and its receptors, LAG3 antagonists or TLR4, TLR7, TLR8, TLR9, STING agonists, etc.), radiotherapy regimens, tumor targeted drugs, and tumor vaccines.

[0042] The fifth aspect of the present invention provides a compound as described in the first aspect of the present invention and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates and deuterated compounds in combination with CAR-T immunotherapy in cancer immunotherapy.

[0043] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0044] After long and in-depth research, the inventors designed and synthesized a new class of HPK1 kinase inhibitors, on the basis of which the inventors completed the present invention.

[0045] the term

[0046] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0047] As used herein, the term "about" when used in reference to a specific recited value means that the value may vary by no more than 5% from the recited value. For example, as used herein, the expression "about 100" includes all values ​​between 95 and 105 (e.g., 95.1, 95.2, 95.3, 95.4, etc.).

[0048] As used herein, the terms "comprising" or "including" may be open, semi-closed, or closed. In other words, the terms also include "consisting essentially of" or "consisting of."

[0049] definition

[0050] As used herein, the term "alkyl" includes straight or branched chain alkyl groups. For example, C1-C8 alkyl groups represent straight or branched chain alkyl groups having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, and the like.

[0051] As used herein, the term "alkenyl" includes straight or branched alkenyl groups. For example, C2-C6 alkenyl refers to a straight or branched alkenyl group having 2 to 6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or the like.

[0052] As used herein, the term "alkynyl" includes straight or branched chain alkynyl groups. For example, C2-C6 alkynyl refers to a straight or branched chain alkynyl group having 2 to 6 carbon atoms, such as ethynyl, propynyl, butynyl, or the like.

[0053] As used herein, the term "C3-C8 cycloalkyl" refers to a cycloalkyl group having 3 to 8 carbon atoms. It can be a monocyclic ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. It can also be a bicyclic ring, such as a bridged ring or a spiro ring.

[0054] As used herein, the term "C1-C8 alkoxy" refers to a straight or branched alkoxy group having 1 to 8 carbon atoms; for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, etc.

[0055] As used herein, the term "3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S, and O" refers to a saturated or partially saturated cyclic group having 3-12 atoms, 1-3 of which are heteroatoms selected from the group consisting of N, S, and O. It may be a monocyclic or polycyclic ring, such as a bridged ring, a spirocyclic ring, or a fused ring. Specific examples include oxetane, azetidine, tetrahydro-2H-pyranyl, piperidinyl, tetrahydrofuranyl, morpholinyl, and pyrrolidinyl.

[0056] As used herein, the term "C6-C 10 The term "aryl" refers to an aromatic group having 6 to 10 carbon atoms, for example, phenyl or naphthyl and the like.

[0057] As used herein, the term "5-10 membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S, and O" refers to a cyclic aromatic group having 5-10 atoms, 1-3 of which are heteroatoms selected from the group consisting of N, S, and O. It may be a monocyclic or condensed ring. Specific examples include pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl and (1,2,4)-triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, and oxazolyl.

[0058] Unless otherwise specified, the groups of the present invention may be substituted by substituents selected from the group consisting of halogen, nitrile, nitro, hydroxy, amino, C1-C6 alkyl-amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C2-C6 alkenyl, halogenated C2-C6 alkynyl, halogenated C1-C6 alkoxy, allyl, benzyl, C6-C6 12 Aryl, C1-C6 alkoxy-C1-C6 alkyl, C1-C6 alkoxy-carbonyl, phenoxycarbonyl, C2-C6 alkynyl-carbonyl, C2-C6 alkenyl-carbonyl, C3-C6 cycloalkyl-carbonyl, C1-C6 alkyl-sulfonyl, etc.

[0059] As used herein, "halogen" or "halogen atom" refers to F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from F, Cl, and Br. "Halogenated" means substituted with an atom selected from F, Cl, Br, and I.

[0060] Unless otherwise specified, the structural formulas described herein are intended to include all isomeric forms (e.g., enantiomers, diastereomers, and geometric isomers (or configurational isomers)): for example, R and S configurations containing asymmetric centers, (Z) and (E) isomers of double bonds, etc. Therefore, individual stereochemical isomers of the compounds of the present invention, or mixtures of such enantiomers, diastereomers, or geometric isomers (or configurational isomers), are within the scope of the present invention. Unless otherwise specified, the absolute stereo configuration of the compounds herein is undetermined and is determined by chiral resolution of racemic compounds. The stereo configurations indicated in the structural formulas and compound names are arbitrarily assigned.

[0061] As used herein, the term "tautomer" refers to structural isomers of different energies that can interconvert across a low energy barrier. For example, proton tautomers (i.e., prototropy) include interconversion via proton migration, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversion via reorganization of some of the bonding electrons.

[0062] As used herein, the term "solvate" refers to a complex in which the compound of the present invention is coordinated with solvent molecules to form a specific ratio.

[0063] Compounds of formula I

[0064] The present invention provides a compound shown in the following formula I:

[0065] in,

[0066] X 1 Selected from the following group: N, CR 3 NR 3 ;

[0067] X 2 Selected from the group consisting of: N, CH;

[0068] X 4 N or CH; Z 1 is N or C; and It is an aromatic ring;

[0069] R 3 Selected from the following group: H, C 1-6 Alkyl, C 3-8 Cycloalkyl (preferably C 3-6 cycloalkyl), a 4-8 membered heterocyclic group containing 1-3 heteroatoms selected from N, O, and S, a 4-8 membered (preferably 5 membered) heteroaryl group containing 1-3 heteroatoms selected from N, O, and S, and the above groups may be further substituted by 1-3 groups selected from the following groups: D, -NR 11 R 12 、-CN、halogen、C 1-6 Alkyl, C 1-6 Alkoxy, -C(O)R 11 、-SO2R 18 ;

[0070] R 11 , R 12 Selected from H, C 1-3 Alkyl, or R 11 、R 12 Together with the atoms to which they are attached, they form a 4-6 membered heterocyclic ring;

[0071] R 18 C 1-3 alkyl;

[0072] R 1 for: Among them, R a and R b Each independently selected from the group consisting of: H, C 1-6 Alkyl, phenyl, 5-7 membered heteroaryl, C 3-8 Cycloalkyl, C 1-6 Alkenyl, benzyl, C 5-8 Spiroalkyl and C 5-8 Bridged cycloalkyl, 4-12 membered (preferably 4-8 membered) heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O, 5-7 membered heteroaromatic substituted C 1-6 Alkyl, or R a and R b The atoms connected to it together form C 3-8 Cycloalkyl;

[0073] And the R a or R b, or R a and R b The ring formed by the atoms connected thereto may be further substituted by 1-3 groups selected from the group consisting of halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl;

[0074] R c Selected from the group consisting of: -NR 16 R 17 ,-OH;R 16 , R 17 Each independently selected from H, C 1-3 alkyl;

[0075] R 2 It has the structure shown below:

[0076] Where n is 0 or 1;

[0077] Y 1 N or CH;

[0078] Each Y 4 and Y 5 Each independently selected from the following group: O, NR 15 、C(R 13 R 14 ); each R 13 , R 14 , R 15 Each independently selected from the following group: H, C 1-6 alkyl;

[0079] R 4 、R 5 Each independently selected from the group consisting of: H, C 1-6 Alkyl (preferably C 1-3 alkyl);

[0080] In the compound of formula I, each chiral center is in R configuration or S configuration.

[0081] Preferably, the X 1 、X 2 、X 4 、Z 1 、Y 1 、Y 4 、Y 5 、R 1 、R 2 、R 3 、R 4 、R 5 、R 11 、R 12 、R13 、R 14 、R 15 、R a 、R b 、R c , m, and n are each independently the corresponding groups in the specific compounds in each embodiment.

[0082] The compounds of the present invention can act as HPK1 kinase inhibitors, and in preferred embodiments, are HPK1 kinase selective inhibitors.

[0083] Preparation of compounds of formula I

[0084] The compounds of formula I of the present invention can be prepared by the following exemplary methods:

[0085] The iodinated parent core compound is subjected to Stille reaction to obtain an alkylated parent core after conversion, which is then coupled with a bicyclic fragment to obtain a compound skeleton, and after modification with substituents, the compounds of formula I of the present invention are obtained.

[0086] Pharmaceutical compositions and methods of administration

[0087] Since the compounds of the present invention have excellent HPK1 kinase inhibitory activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to prevent and / or treat diseases related to HPK1 kinase activity or expression (e.g., cancer).

[0088] The pharmaceutical compositions of the present invention comprise a safe and effective amount of a compound of the present invention and a pharmaceutically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 10-200 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0089] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0090] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral administration, parenteral (intravenous, intramuscular or subcutaneous) injection, and the like.

[0091] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0092] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0093] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0094] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0095] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0096] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0097] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0098] When administered in combination, the pharmaceutical composition may further comprise one or more other pharmaceutically acceptable compounds. One or more of the other pharmaceutically acceptable compounds may be administered simultaneously, separately or sequentially with the compound of the present invention.

[0099] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage. For a 60 kg human, the daily dosage is generally 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage will also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0100] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0101] Synthesis Example

[0102] Synthesis of Intermediate A1: 1-(5-chloro-1-methylpyrazolo[3,4-c]pyridin-3-yl)ethan-1-one

[0103] Step 2: 2-methylpropane-2-yl 3-acetyl-5-chloropyrazolo[3,4-c]pyridine-1-carboxylate

[0104] To a solution of compound 1-1 (20.0 g, 70 mmol) and N,N-diisopropylethylamine (12.1 g, 90 mmol) in tetrahydrofuran (200 mL) was added di-tert-butyl dicarbonate (18.8 g, 90 mol) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction solution was concentrated, and dichloromethane (300 mL) was added. The organic phase was washed with saturated ammonium chloride solution (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to yield compound A1-2 (25.0 g, yield: 92.5%) as a gray solid. MS (ESI): m / z = 379.9 [M+H] + .

[0105] Step 3: 5-chloro-3-(1-ethoxyvinyl)pyrazolo[3,4-c]pyridine-1-carboxylic acid-2-methylprop-2-yl ester

[0106] Under argon, Pd(PPh3)2Cl2 (1.4 g, 2.11 mmol) was added to a solution of compound A1-2 (10.0 g, 26.40 mmol), tributyl(1-ethoxyethylene)tin (9.5 g, 26.40 mmol), and anhydrous N,N-dimethyldiformamide (100 mL). The reaction solution was heated to 85°C and stirred for 16 hours under argon. LCMS confirmed the reaction was complete. The reaction solution was used directly in the next step without further treatment.

[0107] Step 4: 1-(5-chloro-1H-pyrazolo[3,4-c]pyridin-3-yl)ethan-1-one

[0108] To the N,N-dimethyldiformamide reaction solution of the crude product of Compound A1-3, an aqueous solution of hydrochloric acid (2N, 26.4 mL) was added, and the reaction solution was heated to 80°C and stirred for 1 hour. The reaction was detected by LCMS. After the reaction solution was cooled to room temperature, it was filtered, and the filter cake was rinsed three times with ethyl acetate (200 mL). The combined organic phase was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse phase column chromatography (eluted with 20.7% acetonitrile in water) to obtain Compound A1 (6.5 g, yield: 48.8%) as a yellow solid. MS (ESI): m / z=195.9[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ14.62 (s, 1H), 9.01 (d, J = 1.1Hz, 1H), 2.67 (s, 3H).

[0109] Synthesis of Intermediate A2: 5-Chloro-1-methyl-3-(prop-1-en-2-yl)pyrazolo[3,4-c]pyridine

[0110] To a solution of compound 1-2 (1.0 g, 3.41 mmol) and isopropenylboronic acid pinacol ester (0.90 mL, 3.75 mmol) in 1,4-dioxane (15 mL) and water (3 mL) were added [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (249 mg, 0.34 mmol) and potassium phosphate (2.17 g, 10.20 mmol). The reaction mixture was purged with argon and reacted at 85°C for 3 hours. After cooling to room temperature, the reaction mixture was filtered, and the filter cake was washed with ethyl acetate. The filtrate was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-20% ethyl acetate in petroleum ether) to obtain compound A2 (535 mg, yield: 75.6%) as a white solid. MS (ESI): m / z = 208.3 [M+H] + .

[0111] Synthesis of Intermediate A3: Tributyl(2-methylprop-2-enyl)-λ 4 -Stannane

[0112] To a solution of magnesium (358 mg, 14.90 mmol) in tetrahydrofuran (20 mL) at 0°C under argon was added titanium tetrachloride (284 mg, 1.49 mmol) and tributyltin chloride (3.9 g, 11.90 mmol) dropwise. The mixture was slowly allowed to return to room temperature, and 3-bromo-2-methylpropene (2.0 g, 14.90 mmol) was slowly added dropwise. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the mixture was quenched with saturated ammonium chloride solution (10 mL) and extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford compound A3 (4.0 g, yield: 77.5%) as a yellow oil, which was used directly in the next reaction. 1 H NMR (400MHz, CDCl3) δ4.49-4.43(m,2H),1.79(d,J=1.2Hz,2H),1.72-1.70(m,3H),1.56-1.44(m,6H),1.36-1.27(m,6H),0.95-0.6(m,15H).

[0113] Synthesis of Intermediate A4: 2-amino-7-ethyl-7-methyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0114] Step 1: Methyl 2-chloro-6-(((2,4-dimethoxyphenyl)methyl)amino)pyridine-3-carboxylate

[0115] To a mixture of compound A4-0 (10.0 g, 48.50 mmol), cesium fluoride (7.4 g, 48.50 mmol) and N,N-dimethyldiformamide (100 mL) was added 2,4-dimethoxybenzylamine (8.1 g, 48.50 mmol). The reaction mixture was stirred at 80°C for 4 hours and quenched by adding water (10 mL). The mixture was extracted three times with ethyl acetate (50 mL), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-35% ethyl acetate in petroleum ether) to obtain compound A4-1 (4.0 g, yield: 24.5%) as a white solid. MS (ESI): m / z = 337.1 [M+H] + .

[0116] Step 2: 2-chloro-6-[6-(2,4-dimethoxyphenyl)-2,2-dimethyl-4-oxyylidene-5-aza-3-oxahex-5-yl]pyridine-3-carboxylic acid methyl ester

[0117] To a solution of compound A4-1 (1.7 g, 5.06 mmol), triethylamine (1.53 g, 15.20 mmol), and 4-dimethylaminopyridine (123 mg, 1.01 mmol) in dichloromethane (17 mL) was added di-tert-butyl dicarbonate (1.32 g, 6.07 mmol) at room temperature. The reaction mixture was stirred at 65°C for 16 hours. After completion, the reaction was quenched by the addition of water (10 mL) and extracted three times with ethyl acetate (50 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-20% ethyl acetate in petroleum ether) to obtain compound A4-2 (2.0 g, yield: 90.7%) as a white solid. MS (ESI): m / z = 437.2 [M+H] + .

[0118] Step 3: 6-[6-(2,4-dimethoxyphenyl)-2,2-dimethyl-4-oxyidene-5-aza-3-oxahex-5-yl]-2-(2-methylprop-2-enyl)pyridine-3-carboxylic acid methyl ester

[0119] Under argon, Pd(PPh3)2Cl2 (258 mg, 0.37 mmol) was added to a solution of compound A4-2 (1.6 g, 3.67 mmol), compound A3 (2.5 g, 7.34 mmol), lithium chloride (154 mg, 3.67 mmol), and anhydrous N,N-dimethylformamide (16 mL). The reaction solution was heated to 100°C and stirred for 16 hours under argon. After completion, water (5 mL) was added to quench the reaction, and the mixture was extracted three times with ethyl acetate (25 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-20% ethyl acetate in petroleum ether) to obtain compound A4-3 (950 mg, yield: 56.8%) as a colorless oil. MS (ESI): m / z = 457.3 [M+H] + .

[0120] Step 4: 6-[6-(2,4-dimethoxyphenyl)-2,2-dimethyl-4-oxyidene-5-aza-3-oxahexan-5-yl]-2-[(2-methyloxacyclopropyl-2-yl)methyl]pyridine-3-carboxylic acid methyl ester

[0121] To a solution of compound A4-3 (1.0 g, 2.19 mmol) in dichloromethane (10 mL) at 20°C was added m-chloroperbenzoic acid (1.13 g, 6.58 mmol), and the reaction mixture was stirred at 20°C for 2 hours. After the reaction was complete, it was quenched with sodium sulfite solution (10%, 10 mL), and then extracted three times with ethyl acetate (30 mL). The combined organic phases were washed with saturated sodium bicarbonate solution (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound A4-4 (600 mg, yield: 58%) as a colorless oil. MS (ESI): m / z = 473.3 [M+H] + .

[0122] Step 5: (((2,4-dimethoxyphenyl)methyl)(7-ethyl-7-methyl-5-oxyylidene-7,8-dihydro-5H-pyrano[4,3-b]pyridin-2-yl)amino)methane-2-methylpropane-2-yl ester

[0123] Under argon, a solution of cuprous iodide (1.1 g, 5.29 mmol) and anhydrous ether (10 mL) was added to a three-necked reaction flask equipped with a magnetic stirrer. After the solution was cooled to -78°C, a solution of methyllithium in tetrahydrofuran (1.6 M, 6.6 mL, 10.6 mmol) was slowly added dropwise. After the addition was complete, the reaction mixture was slowly warmed to -35°C and stirred for half an hour. The mixture was then cooled to -78°C and a solution of compound A4-4 (500 mg, 5.29 mmol) in ether (1 mL) was slowly added dropwise. The reaction mixture was slowly warmed to 0°C and stirred for 1 hour. After the reaction was complete, a saturated ammonium chloride solution (5 mL) was added to quench the reaction, followed by extraction with ethyl acetate (50 mL) three times. The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 0-30% ethyl acetate in petroleum ether) to obtain compound A4-5 (400 mg, yield: 83%) as a white solid. MS (ESI): m / z = 457.3 [M+H] + .

[0124] Step 6: 2-amino-7-ethyl-7-methyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0125] Compound A4-5 (400 mg, 0.88 mmol) was dissolved in trifluoroacetic acid (5 mL), and the reaction mixture was stirred at 25°C for 2 hours. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse phase column chromatography (eluted with a mixture of 50% acetonitrile and water) to obtain compound A4 (45 mg, yield: 24.9%) as a white solid. MS (ESI): m / z = 207.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.05(d,J=8.0Hz,1H),6.44(d,J=8.0Hz,1H),4.95(brs,2H),3.04(d,J =17.2Hz,1H),2.84(d,J=17.2Hz,1H),1.86-1.71(m,2H),1.41(s,3H),1.01(t,J=8.4Hz,3H).

[0126] Synthesis of Intermediate A6: 1-(5-chloro-1-(oxetan-3-yl)-1H-pyrazolo[3,4-c]pyridin-3-yl)ethan-1-one

[0127] To a solution of compound A1 (293 mg, 1.50 mmol) in N,N-dimethylformamide (5 mL) were added cesium carbonate (1465 mg, 4.49 mmol) and 3-iodooxetane (1378 mg, 7.49 mmol). The reaction mixture was reacted in a microwave reactor at 100°C for 3 hours. After cooling to room temperature, the reaction solution was quenched with water (10 mL) and extracted twice with ethyl acetate (20 mL). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 30% ethyl acetate and petroleum ether) to obtain compound A6 (306 mg, yield: 81.2%) as a white solid. MS (ESI): m / z = 252.3 [M+H] + .

[0128] Synthesis of Intermediate A7: 1-(5-chloro-1-(1-methyl-1H-pyrazol-4-yl)-1H-pyrazolo[3,4-c]pyridin-3-yl)ethan-1-one

[0129] To a solution of compound A1 (293 mg, 1.50 mmol) in 1,4-dioxane (5 mL) were added 1-methyl-4-bromopyrazole (362 mg, 2.25 mmol), cuprous iodide (57 mg, 0.30 mmol), trans-(1R,2R)-N,N'-dimethyl-1,2-cyclohexanediamine (85 mg, 0.60 mmol), and cesium carbonate (1221 mg, 3.75 mmol). The reaction mixture was reacted at 110°C under argon for 16 hours. After cooling to room temperature, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluting with a mixture of 50% ethyl acetate and petroleum ether) to obtain compound A7 (100 mg, yield: 24.2%) as a yellow solid. MS (ESI): m / z = 276.0 [M+H] + .

[0130] Synthesis of Intermediate A9: (5-chloro-1-methylpyrrolo[2,3-c]pyridin-3-yl)(3,3-difluorocyclobutyl)methanone

[0131] Under argon at -10°C, a solution of isopropylmagnesium chloride in tetrahydrofuran (6.8 mL, 13.7 mmol, 2.0 M) was slowly added dropwise to a solution of A9-0 (1.0 g, 3.42 mmol) in dry tetrahydrofuran (10 mL). The reaction mixture was stirred at -10°C for 1 hour, followed by the addition of 3,3-difluoro-N-methoxycyclobutanecarboxamide (1.23 g, 6.84 mmol), and the reaction was stirred for 2 hours. After the reaction was complete, the mixture was returned to room temperature and quenched with saturated ammonium chloride solution (30 mL). The mixture was extracted three times with ethyl acetate (50 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 0-35% ethyl acetate in petroleum ether) to obtain compound A9 (320 mg, yield: 32.8%) as a light yellow solid. MS (ESI): m / z = 284.9 [M+H] + .

[0132] Synthesis of Intermediate A10: 1-(5-chloro-1-methylpyrazolo[3,4-c]pyridin-3-yl)propan-1-one

[0133] Step 1: 5-Chloro-1-methylpyrazolo[3,4-c]pyridine-3-carbonitrile

[0134] Under argon, Pd(dppf)Cl2 (124 mg, 0.17 mmol) and Pd2(dba)3 (156 mg, 0.17 mmol) were added to a mixed solution of compound 1-2 (500 mg, 1.70 mmol) and zinc cyanide (110 mg, 0.94 mmol) in N,N-dimethyldiformamide (3.5 mL) and water (0.5 mL) under argon protection. The reaction mixture was stirred at 80°C for 1 hour. After the reaction was completed, the reaction mixture was slowly added dropwise to ice water (30 mL) and extracted three times with ethyl acetate (50 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 10-20% ethyl acetate in petroleum ether) to obtain compound A10-1 (295 mg, yield: 89.9%) as a white solid. MS (ESI): m / z = 193.0 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.88(s,1H),7.79(s,1H),4.30(s,3H).

[0135] Step 2: 1-(5-chloro-1-methylpyrazolo[3,4-c]pyridin-3-yl)propan-1-one

[0136] Under argon protection and at -78 ° C, to a solution of compound A10-1 (100 mg, 0.52 mmol) and cuprous bromide (37 mg, 0.26 mmol) in anhydrous tetrahydrofuran (2.5 mL) was added ethylmagnesium bromide tetrahydrofuran solution (3.6 mL, 3.6 mmol, 1 M), and the reaction mixture was heated to 50 ° C and stirred for 2 hours. After the reaction was completed, saturated ammonium chloride solution (5 mL) was added to quench the reaction, and the mixture was extracted three times with ethyl acetate (5 mL). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 10-20% ethyl acetate in petroleum ether) to obtain compound A10 (65 mg, yield: 56%) as a white solid. MS (ESI): m / z = 223.9 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.77(d,J=1.0Hz,1H),8.26(d,J=1.0Hz,1H),4.25(s,3H),3.15(q,J=7.4Hz,2H),1.27(t,J=7.0Hz,3H).

[0137] Synthesis of Intermediate A11: 7-(Azidophenylmethyl)-2-chloro-5-methylpyrrolo[3,2-d]pyrimidine

[0138] Step 1: (2-chloro-5H-pyrrolo[3,2-d]pyrimidin-7-yl)phenylmethanol

[0139] To a solution of compound A11-0 (1.0 g, 6.54 mmol) in acetonitrile (10 mL) and water (10 mL) were added benzaldehyde (693 mg, 6.54 mmol) and potassium carbonate (1.35 g, 9.80 mmol) in sequence. The reaction mixture was stirred at 55°C for 72 hours. After completion, the reaction was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse-phase column chromatography (gradient elution with 0-50% acetonitrile in water containing 0.1% ammonium bicarbonate) to obtain compound A11-1 (300 mg, yield: 17.7%) as a white solid. MS (ESI): m / z = 260.1 [M+H] + .

[0140] Using compound A11-1 as an intermediate, the subsequent steps were carried out using a method similar to that in Synthesis Example 6 to obtain compound A11.

[0141] MS (ESI): m / z = 299.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.71(s,1H),7.46(d,J=7.3Hz,2H),7.40-7.38(m,2H),7.34(d,J=7.1Hz,1H),7.28(s,1H),6.26(s,1H),3.88(s,3H).

[0142] Synthesis of intermediates A12 and A13: 3-[1-azido-1-(fluorocyclopentyl)ethyl]-5-chloro-1-methylpyrrolo[2,3-c]pyridine and 3-(1-azido-1-cyclopentyl-2-fluoroethyl)-5-chloro-1-methylpyrrolo[2,3-c]pyridine

[0143] Step 1: 5-chloro-3-(1-cyclopentylideneethyl)-1-methylpyrrolo[2,3-c]pyridine and 5-chloro-3-(1-cyclopentylvinyl)-1-methylpyrrolo[2,3-c]pyridine

[0144] Compound 40-3 (1394 mg, 5 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was added dropwise at room temperature. After the addition was complete, the reaction solution was allowed to react at room temperature for 1 hour. After the reaction was completed, the residue was concentrated under reduced pressure and dissolved in ethyl acetate (20 mL), neutralized with saturated sodium bicarbonate solution, and the organic phase was washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 30% ethyl acetate and petroleum ether) to obtain product A12-1 (761 mg, yield 58.4%) and product A13-1 (70 mg, yield 5.4%), both as yellow oils. A12-1, MS (ESI): m / z = 290.2 [M+H] + .A13-1,MS(ESI):m / z=290.2[M+H] + .

[0145] Step 2: 3-[1-azido-1-(fluorocyclopentyl)ethyl]-5-chloro-1-methylpyrrolo[2,3-c]pyridine

[0146] A12-1 (130 mg, 0.499 mmol), selectfluor (212 mg, 0.598 mmol), sodium azide (78 mg, 1.197 mmol), and sodium carbonate (21 mg, 0.199 mmol) were added to nitromethane (2 mL) at room temperature. The mixture was stirred at 80°C for 2 hours. After the reaction solution was cooled to room temperature, water (10 mL) was added to quench the reaction. The mixture was extracted twice with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluted with a mixture of 20% ethyl acetate in petroleum ether) to obtain product A12 (57 mg, 35.5% yield) as a yellow oil. MS (ESI): m / z = 322.2 [M+H] + .

[0147] 3-(1-azido-1-cyclopentyl-2-fluoroethyl)-5-chloro-1-methylpyrrolo[2,3-c]pyridine

[0148] Compound A13 was obtained using a method similar to that used to synthesize compound A12, starting from A13-1. MS (ESI): m / z = 322.2 [M+H] + .

[0149] The following intermediates were synthesized with reference to the open literature:

[0150] Example 1: 2-(3-(1-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0151] Step 1: 5-chloro-3-iodo-1H-pyrazolo[3,4-c]pyridine

[0152] At room temperature, iodine (3.44 g, 13.54 mmol) was added to a solution of compound 1-0 (1.04 g, 6.77 mmol) and anhydrous potassium carbonate (1.87 g, 13.54 mmol) in acetonitrile (20 mL). The reaction mixture was stirred at room temperature for 13 hours. The reaction was detected by LCMS. After the reaction was completed, the reaction was quenched with sodium sulfite solution (10%, 20 mL), and then extracted three times with dichloromethane (30 mL). The combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 1-1 (1.80 g, yield: 95.1%) as a light yellow solid. MS (ESI): m / z = 279.9 [M+H] + .

[0153] Step 2: 5-chloro-3-iodo-1-methyl-1H-pyrazolo[3,4-c]pyridine

[0154] Under argon protection, iodomethane (1.10 g, 7.74 mmol) was added to a mixture of compound 1-1 (1.80 g, 6.45 mmol), potassium hydroxide (543 mg, 9.68 mmol), and acetone (15 mL). The reaction mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. Dichloromethane (25 mL) was then added and extracted three times. The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 1-35% ethyl acetate in petroleum ether) to obtain compound 1-2 (1.15 g, yield: 60.7%) as a white solid. MS (ESI): m / z = 294.0 [M+H] + . 1 H NMR: (400MHz, CDCl3) δ8.67(s,1H),7.38(s,1H),4.19(s,3H).

[0155] Step 3: 5-chloro-3-(1-ethoxyvinyl)-1-methyl-1H-pyrazolo[3,4-c]pyridine

[0156] Under argon protection, Pd(PPh3)2Cl2 (143 mg, 0.2 mmol) was added to a solution of compound 1-2 (600 mg, 2.04 mmol), tributyl(1-ethoxyethylene)tin (740 mg, 2.04 mmol), and anhydrous N,N-dimethyldiformamide (15 mL). The reaction solution was heated to 60°C and stirred for 16 hours under argon protection. LCMS confirmed the reaction was complete. The reaction solution was used directly in the next step without further treatment. MS (ESI): m / z = 238.1 [M+H] + .

[0157] Step 4: 1-(5-chloro-1-methyl-1H-pyrazolo[3,4-c]pyridin-3-yl)ethan-1-one

[0158] To the N,N-dimethylformamide reaction solution of compound 1-3 (486 mg, 2.04 mmol), hydrochloric acid (2N, 6 mL) aqueous solution and methanol (6 mL) were added sequentially, and the reaction solution was heated to 90 ° C and stirred for 1 hour. The reaction was detected by LCMS. After the reaction solution was cooled to room temperature, saturated sodium bicarbonate solution was added to neutralize it and the pH was adjusted to 7. Ethyl acetate (15 mL) was then added and extracted three times. The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 1-25% ethyl acetate and petroleum ether) to obtain compound 1-4 (335 mg, yield: 82.8%) as a white solid. MS (ESI): m / z=210.1[M+H] + . 1 H NMR (400MHz, CDCl3) δ8.78 (s, 1H), 8.25 (d, J = 0.8Hz, 1H), 4.26 (s, 3H), 2.71 (s, 3H).

[0159] Step 5: 1-(5-chloro-1-methyl-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-cyclopropyl-1-ol

[0160] Under argon protection, compound 1-4 (450 mg, 2.15 mmol) and anhydrous tetrahydrofuran (15 mL) solution were added to a three-necked reaction flask equipped with a magnetic stirrer. After the solution was cooled to -78°C, a tetrahydrofuran solution of cyclopropylmagnesium bromide (1N, 43 mL, 43 mmol) was slowly added dropwise. After the addition was complete, the reaction mixture was slowly heated to 20°C and stirred for 10 hours. 1After the starting material was complete, the reaction was quenched by adding saturated ammonium chloride solution (20 mL), followed by extraction twice with ethyl acetate (10 mL). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to afford the crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 1-40% ethyl acetate in petroleum ether) to afford compound 1-5 (315 mg, yield: 58.4%) as a light yellow oil. 1 H NMR (400MHz, CDCl3) δ8.67(s,1H),7.85(s,1H),4.12(s,3H),1.70(s,3H),1.4-1.35(m,1H),0.53-0.50(m,3H),0.49-0.42(m,1H).

[0161] Step 6: 3-(1-azido-1-cyclopropylethyl)-5-chloro-1-methyl-1H-pyrazolo[3,4-c]pyridine

[0162] Under argon protection, sodium azide (179 mg, 2.75 mmol) was added to a solution of compound 1-5 (315 mg, 1.25 mmol) in anhydrous dichloromethane (6 mL). After cooling to -5 ° C, a solution of trifluoroacetic acid (1.120 g, 10.5 mmol) in dichloromethane (2 mL) was slowly added dropwise to the solution. After the addition was complete, the reaction solution was stirred at 0 ° C for 2 hours. The reaction was detected by LCMS and the raw material conversion was complete. Water (5 mL) and 14% ammonia water (5 mL) solution were added to the reaction solution to quench the reaction. Then dichloromethane was added and extracted (15 mL) twice. The combined organic phase was washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 1-25% ethyl acetate in petroleum ether) to afford compound 1-6 (155 mg, yield: 44.8%) as a light yellow oil. MS (ESI): m / z = 277.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.69(s,1H),7.89(s,1H),4.14(s,3H),1.81(s,3H),1.39-1.36(m,1H),0.57-0.63(m,3H),0.55-0.41(m,1H).

[0163] Step 7: 2-(3-(1-azido-1-cyclopropyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0164] Under argon protection, compound 1-6 (82 mg, 0.30 mmol), 2-amino-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (102 mg, 0.44 mmol), Brettphos Pd G4 (27 mg, 0.030 mmol), Brettphos (24 mg, 0.044 mmol), anhydrous potassium phosphate (157 mg, 0.74 mmol) and anhydrous 1,4-dioxane (1.5 mL) were added sequentially to a dry sealed tube equipped with a stirrer. The final reaction mixture was purged with argon three times and heated to 60 ° C for 16 hours. After the reaction was completed, it was cooled to room temperature and the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 1-35% ethyl acetate and petroleum ether) to obtain compound 1-7 (102 mg, yield: 74.0%) as a light yellow solid. MS (ESI): m / z = 433.5 [M+H] + .

[0165] Step 8: 2-(3-(1-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0166] Under argon protection, platinum dioxide (54 mg, 0.24 mmol) was added to a solution of compound 1-7 (102 mg, 0.24 mmol) in ethyl acetate (5 mL). The mixture was replaced with hydrogen three times and then stirred at room temperature under hydrogen for 1 hour. After the reaction was completed, it was filtered and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by Prep-HPLC to obtain compound 1 (53 mg, yield: 54.7%) as a white solid. MS (ESI): m / z = 407.5 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.78(s,1H),8.62(s,1H),8.05(d,J=8.8Hz,1H),7.15(d,J=8.8Hz,1H),4.14(s, 3H),3.14(s,2H),1.66(s,3H),1.52(s,6H),1.46-1.37(m,1H),0.60-0.49(m,3H),0.47-0.41(m,1H).

[0167] Example 2 and Example 3:

[0168] Example 1 (86 mg, 0.21 mmol) was separated by SFC (chromatographic column: Daicel IBN-5250*4.6mm,5μm; mobile phase A: acetonitrile + 0.2% diethylamine, mobile phase B: isopropanol + 0.2% diethylamine, isocratic elution program: mobile phase A: mobile phase B = 90:10 (V / V), to give (R or S)-3-(3-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrone (1st eluting isomer, 29 mg, yield: 32.6%), and (R or S)-3-(3-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrone (2nd eluting isomer, 29 mg, yield: 32.6%). isomer, 32 mg, yield: 35.9%), all were white solids.

[0169] Example 2: (1 st eluting isomer)(R or S)-3-(3-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrone

[0170] MS (ESI): m / z = 407.5 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.77(s,1H),8.60(s,1H),8.04(d,J=8.8Hz,1H),7.13(d,J=8.8Hz,1H),4.12(s, 3H),3.11(s,2H),1.64(s,3H),1.50(s,6H),1.42-1.34(m,1H),0.60-0.48(m,3H),0.46-0.37(m,1H).

[0171] Example 3: (2 nd eluting isomer)(R or S)-3-(3-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrone

[0172] MS (ESI): m / z = 407.5 [M+H] + . 1H NMR (400MHz, CD3OD) δ8.77(s,1H),8.60(s,1H),8.03(d,J=8.8Hz,1H),7.13(d,J=8.8Hz,1H),4.12(s, 3H),3.11(s,2H),1.64(s,3H),1.50(s,6H),1.42-1.36(m,1H),0.57-0.47(m,3H),0.45-0.37(m,1H).

[0173] Example 4: 2-((3-(1-amino-1-phenylethyl)-1-cyclopropyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0174] Step 1: 1-(5-chloro-1-cyclopropyl-1H-pyrazolo[3,4-c]pyridin-3-yl)ethan-1-one

[0175] To a solution of compound A1 (294 mg, 1.50 mmol) and cyclopropylboronic acid (258 mg, 3.01 mmol) in 1,2-dichloroethane (5 mL) were added copper acetate (301 mg, 1.50 mmol), 2,2'-bipyridine (235 mg, 1.50 mmol), and sodium carbonate (319 mg, 3.01 mmol). The reaction flask was left open and stirred at 70°C for 3 hours. After completion, the reaction solution was cooled to room temperature and quenched with saturated ammonium chloride solution (20 mL). The mixture was extracted with dichloromethane (20 mL). The organic phase was washed twice with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluting with a mixture of 20% ethyl acetate in petroleum ether) to obtain compound 4-1 (228 mg, yield: 64.4%) as a white solid. MS (ESI): m / z = 236.3 [M+H] + .

[0176] Step 2: 1-(5-chloro-1-cyclopropyl-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-phenylethane-1-ol

[0177] Under argon protection, phenylmagnesium bromide (17 mL, 17 mmol) was added dropwise to a solution of compound 4-1 (200 mg, 0.85 mmol) in tetrahydrofuran (3 mL) at -78°C. After the addition was complete, the reaction solution was stirred at -78°C for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution (10 mL) and extracted twice with ethyl acetate (20 mL). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow crude product, compound 4-2. The crude product was used directly in the next step. MS (ESI): m / z = 314.4 [M+H] + .

[0178] Step 3: 3-(1-azido-1-phenylethyl)-5-chloro-1-cyclopropyl-1H-pyrazolo[3,4-c]pyridine

[0179] The crude compound 4-2 obtained in the previous step was dissolved in dichloromethane (2 mL), and trimethylsilyl azide (195 mg, 1.70 mmol) was added. Boron trifluoride etherate (241 mg, 1.70 mmol) was added dropwise to the mixture at 0°C under argon protection. After the addition was complete, the reaction solution was warmed to room temperature and the reaction continued for 1 hour. After the reaction was completed, the reaction solution was quenched with water (10 mL) and extracted twice with dichloromethane (10 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 20% ethyl acetate in petroleum ether) to obtain compound 4-3 (140 mg, yield: 48.8%) as a yellow solid. MS (ESI): m / z = 339.4 [M+H] + .

[0180] Step 4: 2-((3-(1-azido-1-phenylethyl)-1-cyclopropyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0181] Potassium phosphate (244 mg, 1.15 mmol) was added to a solution of compound 4-3 (130 mg, 0.38 mmol) and compound A14 (81 mg, 0.42 mmol) in anhydrous 1,4-dioxane (3 mL). BrettPhos-Pd-G4 (35 mg, 0.038 mmol) and BrettPhos (21 mg, 0.038 mmol) were then added under argon. The reaction mixture was stirred at 100°C under argon for 6 hours. After cooling to room temperature, the mixture was quenched with water (10 mL) and extracted twice with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluting with a mixture of 50% ethyl acetate in petroleum ether) to obtain compound 4-4 (118 mg, yield: 62.2%) as a yellow solid. MS (ESI): m / z = 495.5 [M+H] + .

[0182] Step 5: 2-((3-(1-amino-1-phenylethyl)-1-cyclopropyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0183] To a solution of compound 1-4 (118 mg, 0.239 mmol) in ethyl acetate (3 mL) was added platinum dioxide (54 mg, 0.24 mmol). The reaction mixture was purged with hydrogen three times and stirred under hydrogen at room temperature for 1 hour. The reaction mixture was filtered, the filter cake washed with ethyl acetate, and the filtrate concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by Prep-HPLC (gradient elution with 20-90% acetonitrile in water containing 0.1% ammonium bicarbonate) to obtain compound 4 (56 mg, yield: 50.1%) as a white solid. MS (ESI): m / z = 469.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.19(s,1H),8.86(d,J=1.0Hz,1H),8.24(s,1H),7.92(d,J=8.7Hz,1H),7.49(d,J=7.4Hz ,2H),7.30-7.28(m,2H),7.20-7.18(m,1H),7.10(d,J=8.7Hz,1H),3.85-3.74(m,1H),3.04-2.83(m,2H),2.43(br s,2H),1.87(s,3H),1.48(s,3H),1.44(s,3H),1.26-1.12(m,4H).

[0184] Example 5: 2-((3-(2-amino-4,4,4-trifluorobutan-2-yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0185] Step 1: 3-(2-azido-4,4,4-trifluorobutan-2-yl)-5-chloro-1-methyl-1H-pyrazolo[3,4-c]pyridine

[0186] To a solution of compound A2 (493 mg, 1.50 mmol) and tetraethylcyanocopper hexafluorophosphate (37 mg, 0.10 mmol) in N,N-dimethylacetamide (5 mL) were added compound 5-0 (207 mg, 0.10 mmol) and trimethylsilylazide (229 mg, 2.0 mmol). The reaction mixture was stirred at room temperature for 16 hours under argon. After completion, the reaction mixture was diluted with ethyl acetate (20 mL) and washed twice with water (10 mL) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluting with a mixture of 30% ethyl acetate and petroleum ether) to obtain compound 5-1 (232 mg, yield: 73.0%) as a yellow solid. MS (ESI): m / z = 319.2 [M+H] + .

[0187] Using compound 5-1 as an intermediate, compound 5 was obtained in the subsequent steps using a method similar to that of Example 4.

[0188] 2-((3-(2-amino-4,4,4-trifluorobutan-2-yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0189] MS (ESI): m / z = 449.4 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.25(s,1H),8.91(d,J=1.0Hz,1H),8.73(s,1H),7.99(d,J=8 .7Hz,1H),7.25(d,J=8.8Hz,1H),4.10(s,3H),3.07(s,2H),3.02-2.86(m,2H),2.35(br s,2H),1.69(s,3H),1.44(s,6H).

[0190] Example 6: 2-((3-(1-amino-1-cyclopentylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0191] Step 1: 5-Chloro-N-methoxy-N-methyl-1H-pyrazolo[3,4-c]pyridine-3-carboxamide

[0192] To a solution of 5-chloro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid (530 mg, 2.68 mmol) and dimethylhydroxylamine hydrochloride (523 mg, 5.37 mmol) in N,N-dimethylformamide (6 mL) at 0°C were added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (564 mg, 2.95 mmol) and 4-dimethylaminopyridine (361 mg, 2.95 mmol) in portions. The reaction mixture was stirred at 0°C for 2 hours. The reaction system was quenched with water (10 mL) and extracted twice with ethyl acetate (20 mL). The organic phases were combined, washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by reverse phase column chromatography (gradient elution with 5-40% methanol in water containing 0.1% ammonium bicarbonate) to obtain compound 6-1 (333 mg, yield: 51.6%) as a white solid. MS (ESI): m / z = 241.0 [M+H] + .

[0193] Step 2: (5-chloro-1H-pyrazolo[3,4-c]pyridin-3-yl)(cyclopentyl)methanone

[0194] To a solution of compound 6-1 (503 mg, 2.09 mmol) in tetrahydrofuran (10 mL) was added dropwise a solution of cyclopentylmagnesium chloride in tetrahydrofuran (10.5 mL, 21 mmol, 2 M) at 0°C. After the addition was complete, the reaction mixture was stirred at 0°C for 1 hour. The mixture was quenched with saturated ammonium chloride solution (10 mL) and extracted twice with ethyl acetate (20 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 35% ethyl acetate in petroleum ether) to obtain compound 6-2 (303 mg, yield: 58.1%) as a yellow solid. MS (ESI): m / z = 250.0 [M+H] + .

[0195] Step 3: (5-chloro-1-methyl-1H-pyrazolo[3,4-c]pyridin-3-yl)(cyclopentyl)methanone

[0196] To a solution of compound 6-2 (303 mg, 1.21 mmol) in N,N-dimethylformamide (3 mL) at 0°C was added cesium carbonate (475 mg, 1.46 mmol), followed by dropwise addition of iodomethane (207 mg, 1.46 mmol). After the addition was complete, the reaction mixture was stirred at room temperature for 2 hours. Ethyl acetate (20 mL) was added to the reaction system for dilution, and the mixture was washed twice with water (10 mL) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 30% ethyl acetate and petroleum ether) to obtain compound 6-3 (253 mg, yield: 79.1%) as a white solid. MS (ESI): m / z = 263.9 [M+H] + .

[0197] Step 4: 1-(5-chloro-1-methyl-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-cyclopentylethane-1-ol

[0198] Under argon protection, methylmagnesium bromide (5 mL, 5 mmol) was added dropwise to a solution of compound 6-3 (253 mg, 0.96 mmol) in tetrahydrofuran (2 mL) at -78°C. After the addition was complete, the reaction solution was stirred at -78°C for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution (10 mL) and extracted twice with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude compound 6-4. The crude product was used directly in the next step. MS (ESI): m / z = 279.9 [M+H] + .

[0199] Step 5: 3-(1-azido-1-cyclopentylethyl)-5-chloro-1-methyl-1H-pyrazolo[3,4-c]pyridine

[0200] The crude compound 6-4 obtained in the previous step was dissolved in dichloromethane (5 mL), followed by the addition of trimethylsilyl azide (731 mg, 6.36 mmol). Boron trifluoride etherate (903 mg, 6.36 mmol) was added dropwise to the mixture at 0°C under argon protection. After the addition was complete, the reaction solution was warmed to room temperature and the reaction continued for 16 hours. After the reaction was complete, the reaction solution was quenched with saturated ammonium chloride (10 mL) and extracted twice with dichloromethane (10 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 25% ethyl acetate in petroleum ether) to obtain compound 6-5 (279 mg, yield: 96.4%) as a colorless oil. MS (ESI): m / z = 304.9 [M+H] + .

[0201] Step 6: 2-((3-(1-azido-1-cyclopentylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0202] Potassium phosphate (583 mg, 2.75 mmol) was added to a solution of compound 6-5 (279 mg, 0.92 mmol) and compound A14 (194 mg, 1.01 mmol) in anhydrous 1,4-dioxane (3 mL). BrettPhos-Pd-G4 (84 mg, 0.092 mmol) and BrettPhos (49 mg, 0.092 mmol) were added under argon. The reaction mixture was stirred at 70°C under argon for 16 hours. After cooling to room temperature, the reaction mixture was quenched with water (10 mL) and extracted twice with ethyl acetate (10 mL). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 50% ethyl acetate in petroleum ether) to obtain compound 6-6 (328 mg, yield: 77.8%) as a yellow solid. MS (ESI): m / z = 461.0 [M+H] + .

[0203] Step 7: 2-((3-(1-amino-1-cyclopentylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0204] To a solution of compound 6-6 (328 mg, 0.71 mmol) in ethyl acetate (3 mL) was added platinum dioxide (162 mg, 0.71 mmol). The reaction mixture was purged with hydrogen three times and stirred at room temperature under hydrogen for 1 hour. The reaction mixture was filtered, the filter cake washed with ethyl acetate, and the filtrate concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of dichloromethane containing 5% methanol) to obtain compound 6 (260 mg, yield: 83.2%) as a white solid. MS (ESI): m / z = 435.0 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.79(s,1H),8.61(s,1H),8.06(d,J=8.8Hz,1H),7.15(d,J=8.8Hz,1H),4.14(s,3H) ,3.12(s,2H),2.67-2.58(m,1H),1.80-1.74(m,1H),1.72(s,3H),1.64-1.48(m,12H),1.45-1.37(m,1H).

[0205] Example 7: (R or S)-2-((3-(1-amino-1-cyclopentylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0206] Compound 6 was subjected to chiral separation to obtain compound 7 as a white solid. MS (ESI): m / z = 435.0 [M+H] + .

[0207] Example 8: 7,7-dimethyl-2-({3-[1-(methylamino)-1-phenylethyl]-1-(trideuteriomethyl)pyrazolo[3,4-c]pyridin-5-yl}amino)-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0208] Step 1: 1-[5-chloro-1-(trideuteriomethyl)pyrazolo[3,4-c]pyridin-3-yl]ethan-1-one

[0209] Under ice bath, to a mixture of compound A1 (420 mg, 2.15 mmol) and anhydrous cesium carbonate (770 mg, 2.36 mmol) in anhydrous N, N-dimethylformamide (8 mL), a solution of deuterated iodomethane (342 mg, 2.36 mmol) in anhydrous N, N-dimethylformamide (1 mL) was slowly added dropwise, and the mixture was stirred under ice bath for 2 hours. After the reaction was completed, the reaction solution was quenched with water (10 mL), extracted three times with ethyl acetate (30 mL), the organic phases were combined, washed three times with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 15% ethyl acetate and petroleum ether) to obtain compound 8-1 (295 mg, yield: 59.4%) as a light yellow solid. MS (ESI): m / z=213.3[M+H] + .

[0210] Using compound 8-1 as an intermediate, the subsequent steps were similar to those in Example 4 to obtain compound 8-4.

[0211] Step 5: ((1-(5-chloro-1-(trideuteriomethyl)pyrazolo[3,4-c]pyridin-3-yl)-1-phenylethyl)amino)methanoic acid-2-methylprop-2-yl ester

[0212] To a solution of compound 8-4 (150 mg, 0.52 mmol) in anhydrous methanol (4 mL) at room temperature was added di-tert-butyl dicarbonate (170 mg, 0.78 mmol), and the mixture was stirred at room temperature for 16 hours. Upon completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 10% ethyl acetate and petroleum ether) to obtain compound 8-5 (195 mg, yield: 96.6%) as a light yellow solid. MS (ESI): m / z = 390.2 [M+H] + .

[0213] Step 6: ((1-(5-chloro-1-(trideuteriomethyl)pyrazolo[3,4-c]pyridin-3-yl)-1-phenylethyl)(methyl)amino)methanoic acid-2-methylprop-2-yl ester

[0214] Under ice-bath conditions, sodium hydride (50 mg, 1.25 mmol) was added portionwise to a solution of compound 8-5 (195 mg, 0.50 mmol) in dry N,N-dimethylformamide (5 mL). The mixture was stirred under ice-bath conditions for 15 minutes. A solution of iodomethane (355 mg, 2.51 mmol) in N,N-dimethylformamide (2 mL) was then slowly added dropwise, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with saturated aqueous ammonium chloride (10 mL) and extracted twice with ethyl acetate (15 mL). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 10% ethyl acetate in petroleum ether) to obtain compound 8-6 (180 mg, yield: 89.1%) as a light yellow solid. MS (ESI): m / z = 404.2 [M+H] + .

[0215] Using compound 8-6 as an intermediate, the subsequent steps were similar to those in Example 4 to obtain compound 8-7.

[0216] Step 8: Bis(1-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-N-(1-{5-[(7,7-dimethyl-5-oxyylidene-7,8-dihydro-5H-pyrano[4,3-b]pyridin-2-yl)amino]-1-(trideuteriomethyl)pyrazolo[3,4-c]pyridin-3-yl}-1-phenylethyl)-N-methylmethaneamide)

[0217] At room temperature, tert-butyldimethylsilyl trifluoromethanesulfonate (156 mg, 0.59 mmol) was added dropwise to a solution of compound 8-7 (110 mg, 0.20 mmol) and 2,6-lutidine (84 mg, 0.79 mmol) in 1,2-dichloroethane (2 mL). After the addition was complete, the reaction solution was allowed to react at 60°C for 1 hour. After cooling to room temperature, the reaction solution was quenched with saturated ammonium chloride (10 mL) and extracted twice with ethyl acetate (10 mL). The organic phases were combined, washed twice with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude compound 8-8. The crude product was used directly in the next step. MS (ESI): m / z = 618.3 [M+H] + .

[0218] Step 9: 7,7-dimethyl-2-({3-[1-(methylamino)-1-phenylethyl]-1-(trideuteriomethyl)pyrazolo[3,4-c]pyridin-5-yl}amino)-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0219] The crude compound 8-8 obtained in the previous step was dissolved in tetrahydrofuran (2 mL), and then pyridine hydrofluoride (97 mg, 0.98 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was separated and purified by reverse-phase column chromatography (gradient elution with 5-50% acetonitrile in water containing 0.1% ammonium bicarbonate) to obtain compound 8 (63 mg, yield: 70.0%) as a white solid. MS (ESI): m / z = 460.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.15(s,1H),8.86(d,J=1.1Hz,1H),8.25(s,1H),7.90(d,J=8.7Hz,1H),7.51-7.42(m,2H),7.30-7.28(m,2H),7 .21-7.19(m,1H),7.06(d,J=8.7Hz,1H),2.87(dd,J=49.6,16.8Hz,2H),2.49(s,1H),2.11(s,3H),1.82(s,3H),1.45(s,3H),1.41(s,3H).

[0220] Example 11: 2-((3-(1-amino-1-cyclopentylethyl)-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0221] Compound 11-3 was obtained using a method similar to Example 6.

[0222] Step 4: 1-(5-chloro-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-cyclopentylethane-1-amine

[0223] To a solution of compound 11-3 (106 mg, 0.40 mmol) in N,N-dimethylformamide (2 mL) at 0°C was added cesium carbonate (196 mg, 0.60 mmol), followed by the dropwise addition of 2,2-difluoroethyl trifluoromethanesulfonate (129 mg, 0.60 mmol). After the addition was complete, the reaction mixture was warmed to room temperature and stirred for 2 hours. Ethyl acetate (20 mL) was added to the reaction system for dilution, and the mixture was washed twice with water (10 mL) and saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of dichloromethane containing 5% methanol) to obtain compound 11-4 (67 mg, yield: 50.9%) as a colorless oil. MS (ESI): m / z = 312.0 [M-NH2] + .

[0224] Step 5: 2-((3-(1-amino-1-cyclopentylethyl)-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0225] Potassium phosphate (144 mg, 0.68 mmol) was added to a solution of compound 11-4 (67 mg, 0.23 mmol) and compound A14 (43 mg, 0.23 mmol) in anhydrous 1,4-dioxane (1.5 mL). BrettPhos-Pd-G4 (21 mg, 0.023 mmol) and BrettPhos (12 mg, 0.023 mmol) were added under argon. The reaction mixture was stirred at 70°C under argon for 16 hours. After cooling to room temperature, the mixture was diluted with ethyl acetate (20 mL). The organic phase was washed with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by Prep-HPLC (gradient elution with 30-90% acetonitrile in water containing 0.1% ammonium bicarbonate) to obtain compound 11 (45 mg, yield: 44.0%) as a white solid. MS (ESI): m / z = 485.3 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.23(s,1H),8.89(s,1H),8.74(s,1H),7.96(d,J=8.7Hz,1H),7.22(d,J=8.9Hz,1 H),6.56-6.25(m,1H),4.99-4.91(m,2H),3.03(s,2H),2.47-2.39(m,1H),1.93(s,2H),1.64-1.29(m,17H).

[0226] Example 12: (R or S)-2-((3-(1-amino-1-cyclopentylethyl)-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0227] Compound 11 (40 mg) was subjected to chiral separation to give compound 12 (15 mg, yield: 37.5%) as a white solid. MS (ESI): m / z = 485.2 [M+H] + .

[0228] Example 13: 2-((3-(1-amino-1-cyclopentylethyl)-1-(difluoromethyl)-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0229] Step 1: 1-(5-chloro-1-(difluoromethyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-1-cyclopentylethane-1-ol

[0230] To a solution of compound 11-1 (176 mg, 0.66 mmol) in acetonitrile (2 mL) at 0°C was added cesium fluoride (302 mg, 1.99 mmol), followed by the dropwise addition of diethyl bromofluoromethylphosphonate (265 mg, 0.99 mmol). After the addition was complete, the reaction mixture was stirred at 0°C for 1 hour, quenched with water (10 mL), extracted with ethyl acetate (20 mL), and the organic phase washed twice with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 50% ethyl acetate in petroleum ether) to obtain compound 13-1 (35 mg, yield: 16.7%) as a colorless oil. MS (ESI): m / z = 316.0 [M+H] + .

[0231] The subsequent steps were carried out with reference to Example 6, and compound 13 was obtained using a similar method.

[0232] MS (ESI): m / z = 471.1 [M+H] + . 1 H NMR (400MHz, CD3OD) δ9.06(s,1H),8.87(s,1H),8.13(d,J=8.7Hz,1H),7.92(t,J=58.8Hz,1H),7.23(d,J =8.7Hz,1H),3.22-3.11(m,2H),2.83-2.72(m,1H),2.02(s,3H),1.97-1.86(m,1H),1.79-1.44(m,13H).

[0233] Example 14: 2-(3-(2-aminopropyl)-1-methylpyrazolo[3,4-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydroquinolin-5(6H)one

[0234] Step 1: 2-{[3-(2-azidoprop-2-yl)-1-methylpyrazolo[3,4-c]pyridin-5-yl]amino}-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-5-one

[0235] Under argon protection, to a mixture of compound 15-2 (80 mg, 0.32 mmol), 6-amino-3,3-dimethyl-3,4-dihydronaphthalen-1(2H)one (73 mg, 0.38 mmol), Brettphos Pd G4 (29 mg, 0.03 mmol), Brettphos (17 mg, 0.03 mmol) and potassium sulfate (203 mg, 0.96 mmol), anhydrous 1,4-dioxane (0.8 mL) was added. The reaction mixture was heated to 70 ° C and stirred for 3 hours under argon protection. After the reaction was cooled to room temperature, water (20 mL) was added to quench the mixture and extracted with ethyl acetate (30 mL). The organic phase was washed with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse-phase column chromatography (gradient elution with 0-50% acetonitrile in water containing 0.1% ammonium bicarbonate) to afford compound 14-1 (120 mg, yield: 83.6%) as a yellow solid. MS (ESI): m / z = 405.4 [M+H] + .

[0236] Step 2: 2-{[3-(2-aminopropyl-2-yl)-1-methylpyrazolo[3,4-c]pyridin-5-yl]amino}-7,7-dimethyl-5,6,7,8-tetrahydroquinolin-5-one

[0237] Under argon protection, platinum dioxide (40 mg, 0.18 mmol) was added to a solution of compound 14-1 (72 mg, 0.18 mmol) in ethyl acetate (2 mL). The mixture was replaced with hydrogen three times and then stirred at room temperature under hydrogen for 1 hour. After the reaction was completed, the mixture was filtered and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by Prep-HPLC to obtain compound 14 (16 mg, yield: 23.7%) as a white solid. MS (ESI): m / z = 379.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.15(s,1H),8.86(s,1H),8.78(s,1H),7.94(d,J=8.8Hz,1H), 7.17(d,J=8.8Hz,1H),4.06(s,3H),2.86(s,2H),2.45(s,2H),1.57(s,6H),1.05(s,6H).

[0238] Example 15: 2-(3-(2-amino-2-yl)-5-(7,7-dimethyl-5-oxo-7,8-dihydro-5H-pyrano[4,3-b]pyridin-2-yl)amino)-1H-pyrazolo[3,4-c]pyridin-1-yl)acetonitrile

[0239] Step 1: 2-(5-chloropyrazolo[3,4-c]pyridin-3-yl)propanol

[0240] Under argon, compound A1 (1000 mg, 5.11 mmol) and anhydrous tetrahydrofuran (4 mL) were added to a three-necked reaction flask equipped with a magnetic stirrer. After the solution was cooled to -78°C, a solution of methyllithium in tetrahydrofuran (2N, 4.1 mL, 8.16 mmol) was slowly added dropwise. After the addition was complete, the reaction mixture was stirred at -78°C for 5 hours. The reaction was quenched by the addition of hydrochloric acid (1N, 10 mL), and then extracted twice with ethyl acetate (100 mL). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-60% ethyl acetate in petroleum ether) to obtain compound 15-1 (680 mg, yield: 62.9%) as a light yellow solid. MS (ESI): m / z = 212.3 [M+H] + .

[0241] Step 2: 3-(2-azidopropan-2-yl)-5-chloropyrazolo[3,4-c]pyridine

[0242] Under argon, trimethylsilyl azide (555 mg, 4.82 mmol) was added to a solution of compound 15-1 (680 mg, 3.21 mmol) in anhydrous dichloromethane (5 mL). After cooling to 0°C, boron trifluoride etherate (912 mg, 6.42 mmol) was slowly added dropwise. After the addition was complete, the reaction solution was stirred at room temperature for 3 hours. Water (10 mL) and 14% aqueous ammonia (5 mL) were added to quench the reaction, followed by extraction with dichloromethane (15 mL) twice. The combined organic phases were washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-25% ethyl acetate in petroleum ether) to obtain compound 15-2 (630 mg, yield: 82.9%) as a light yellow solid. MS (ESI): m / z = 237.3 [M+H] + .

[0243] Step 3: 2-(3-(2-azidopropane-2-yl)-5-chloro-1H-pyrazolo[3,4-c]pyridin-1-yl)acetonitrile

[0244] Under argon protection, 2-bromoacetonitrile (253 mg, 2.11 mmol) was added to a mixture of compound 15-2 (100 mg, 0.42 mmol), cesium carbonate (275 mg, 0.85 mmol), and N,N-dimethylformamide (1 mL). The reaction mixture was stirred at room temperature for 30 minutes and quenched by adding water (10 mL). The mixture was extracted three times with ethyl acetate (25 mL), and the combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-35% ethyl acetate in petroleum ether) to obtain compound 15-3 (80 mg, yield: 68.7%) as a colorless oil. MS (ESI): m / z = 276.3 [M+H] + .

[0245] Step 4: 2-(3-(2-azidopropane-2-yl)-5-(7,7-dimethyl-5-oxo-7,8-dihydro-5H-pyrano[4,3-b]pyridin-2-yl)amino)-1H-pyrazolo[3,4-c]pyridin-1-yl)acetonitrile

[0246] Under argon protection, compound 15-3 (47 mg, 0.17 mmol), compound A14 (33 mg, 0.17 mmol), Brettphos Pd G4 (16 mg, 0.02 mmol), Brettphos (12 mg, 0.026 mmol), anhydrous potassium phosphate (91 mg, 0.43 mmol), and anhydrous 1,4-dioxane (0.3 mL) were added sequentially to a dry, sealed tube equipped with a stirrer. The reaction mixture was purged with argon three times and heated to 60°C for 10 hours. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 0-65% ethyl acetate in petroleum ether) to obtain compound 15-4 (63 mg, yield: 85.7%) as a white solid. MS (ESI): m / z = 432.5 [M+H] + .

[0247] Step 5: 2-(3-(2-amino-2-yl)-5-(7,7-dimethyl-5-oxo-7,8-dihydro-5H-pyrano[4,3-b]pyridin-2-yl)amino)-1H-pyrazolo[3,4-c]pyridin-1-yl)acetonitrile

[0248] Under argon protection, acetic acid (56 mg, 0.093 mmol) was added to a solution of compound 15-4 (40 mg, 0.093 mmol) and zinc powder (61 mg, 0.093 mmol) in tetrahydrofuran (0.3 mL). The mixture was stirred at room temperature for 4 hours. After the reaction was completed, the mixture was filtered and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by Prep-HPLC to obtain compound 15 (2 mg, yield: 5.3%) as a white solid. MS (ESI): m / z = 406.5 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.90(s,1H),8.72(s,1H),8.08(d,J=8.8Hz,1H),7.22(d,J=8.8Hz,1H),3.15(s,2H),1.74(s,6H),1.53(s,6H).

[0249] Example 16: 2-(3-(1-amino-1-cyclopropylethyl)-1-(1-ethylazetidin-3-yl)-1H-pyrazolo[3,4-c]pyridine-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0250] Step 1: tert-Butyl 3-(3-acetyl-5-chloro-1H-pyrazolo[3,4-c]pyridin-1-yl)azetidine-1-carboxylate

[0251] Under argon protection, tert-butyl 3-iodoazetidine-1-carboxylate (782 mg, 2.76 mmol) was added to a mixture of compound A1 (300 mg, 1.53 mmol), cesium carbonate (600 mg, 1.84 mmol) and N, N-dimethylformamide (3 mL). The reaction mixture was stirred at 90 ° C for 16 hours, and water (10 mL) was added to quench the reaction after cooling. It was extracted three times with dichloromethane (25 mL), and the combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-35% ethyl acetate and petroleum ether) to obtain compound 16-1 (420 mg, yield: 78.1%) as a brown solid. MS (ESI): m / z = 351.1 [M + H] + .

[0252] Step 2: tert-Butyl 3-(5-chloro-3-(1-cyclopropyl-1-hydroxyethyl)-1H-pyrazolo[3,4-c]pyridin-1-yl)azetidine-1-carboxylate

[0253] Under argon, compound 16-1 (370 mg, 1.06 mmol) and anhydrous tetrahydrofuran (2 mL) were added to a three-necked reaction flask equipped with a magnetic stirrer. After the solution was cooled to -78°C, a solution of cyclopropylmagnesium bromide in tetrahydrofuran (1N, 10.6 mL, 10.6 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred for 2 hours. The reaction was quenched with saturated ammonium chloride solution (20 mL). The mixture was extracted twice with ethyl acetate (10 mL). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-40% ethyl acetate in petroleum ether) to obtain compound 16-2 (400 mg, yield: 96.5%) as a brown oil. MS (ESI): m / z = 393.1 [M+H] + .

[0254] Step 3: tert-Butyl 3-(3-(1-azido-1-cyclopropylethyl)-5-chloro-1H-pyrazolo[3,4-c]pyridin-1-yl)azetidine-1-carboxylate

[0255] Under argon, trimethylsilyl azide (176 mg, 1.53 mmol) was added to a solution of compound 16-2 (400 mg, 1.02 mmol) in anhydrous dichloromethane (4 mL). After cooling to 0°C, boron trifluoride etherate (289 mg, 2.04 mmol) was slowly added dropwise to the solution. After the addition was complete, the reaction solution was stirred at room temperature for 3 hours. Water (10 mL) and 14% aqueous ammonia (5 mL) were added to quench the reaction, followed by extraction with dichloromethane (15 mL) twice. The combined organic phases were washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-25% ethyl acetate in petroleum ether) to obtain compound 16-3 (182 mg, yield: 42.8%) as a light yellow solid. MS (ESI): m / z = 418.2 [M+H] + .

[0256] Step 4: 3-(1-azido-1-cyclopropylethyl)-5-(7,7-dimethyl-5-oxo-7,8-dihydro-5H-pyrano[4,3-b]pyridin-2-yl)amino)-1H-pyrazolo[3,4-c]pyridin-1-yl)azetidine-1-carboxylic acid tert-butyl ester

[0257] Under argon protection, compound 16-3 (180 mg, 0.43 mmol), 2-amino-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (83 mg, 0.43 mmol), Brettphos Pd G4 (40 mg, 0.043 mmol), Brettphos (31 mg, 0.065 mmol), anhydrous potassium phosphate (229 mg, 1.08 mmol) and anhydrous 1,4-dioxane (1.5 mL) were added sequentially to a dry sealed tube equipped with a stirrer. The reaction mixture was replaced with argon three times and heated to 60 ° C for 10 hours. After the reaction was completed, it was cooled to room temperature and the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-65% ethyl acetate in petroleum ether) to obtain compound 16-4 (100 mg, yield: 40.5%) as a yellow solid. MS (ESI): m / z = 574.3 [M+H] + .

[0258] Step 5: tert-Butyl 3-(3-(1-amino-1-cyclopropylethyl)-5-(7,7-dimethyl-5-oxo-7,8-dihydro-5H-pyrano[4,3-b]pyridin-2-yl)amino)-1H-pyrazolo[3,4-c]pyridin-1-yl)azetidine-1-carboxylate

[0259] Under argon protection, sodium borohydride (2.0 mg, 0.052 mmol) was added to a solution of compound 16-4 (30 mg, 0.052 mmol) in dichloromethane (0.5 mL) and stirred at room temperature for 30 minutes. Water (10 mL) and dichloromethane (10 mL) were added to quench the reaction. The mixture was extracted with dichloromethane (5 mL) twice, and the combined organic phase was washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-10% methanol in dichloromethane) to obtain compound 16-5 (12 mg, yield: 41.9%) as a yellow solid. MS (ESI): m / z = 548.3 [M+H] + .

[0260] Step 6: 2-(3-(1-amino-1-cyclopropylethyl)-1-azetidin-3-yl)-1H-pyrazolo[3,4-c]pyridine-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0261] Under argon, trifluoroacetic acid (0.1 mL, 0.037 mmol) was added to a solution of compound 16-5 (20 mg, 0.037 mmol) in dichloromethane (0.2 mL) and stirred at room temperature for 30 minutes. After completion of the reaction, the mixture was concentrated under reduced pressure to afford compound 16-6 (16 mg, yield: 98.2%) as a yellow oil, which was used directly in the next reaction. MS (ESI): m / z = 448.3 [M+H] + .

[0262] Step 7: 2-(3-(1-amino-1-cyclopropylethyl)-1-(1-ethylazetidin-3-yl)-1H-pyrazolo[3,4-c]pyridine-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0263] Under argon, acetaldehyde (16 mg, 0.036 mmol) and sodium cyanoborohydride (11 mg, 0.18 mmol) were added to a solution of compound 16-6 (16 mg, 0.036 mmol) in methanol (0.3 mL) and stirred at 0°C for 30 minutes. After completion of the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by Prep-HPLC to obtain compound 16 (3 mg, yield: 17.7%) as a white solid. MS (ESI): m / z = 476.3 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.92(s,1H),8.74(s,1H),8.09(d,J=8.8Hz,1H),7.16(d,J=8.8Hz,1H),5.86-5.80(m,1H),4.57-4.50 (m,2H),4.41-4.34(m,2H),3.18(s,2H),1.80(s,2H),1.54(s,6H),1.30-1.25(m,3H),0.91-0.71(m,3H),0.67-0.65(m,1H).

[0264] Example 17: 2-{[3-(1-amino-1-phenylethyl)-1-(2-methoxyethyl)pyrazolo[3,4-c]pyridin-5-yl]amino}-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0265] Step 1: 1-[5-chloro-1-(2-methoxyethyl)pyrazolo[3,4-c]pyridin-3-yl]ethan-1-one

[0266] Under argon protection, compound A1 (450 mg, 2.30 mmol), 2-bromoethyl methyl ether (351 mg, 2.53 mmol), and cesium carbonate (1499 mg, 4.60 mmol) were mixed evenly and N,N-dimethylformamide (5 mL) was added. The mixture was reacted at room temperature under argon protection for 16 hours. After completion of the reaction, it was quenched with water (30 mL). The reaction solution was extracted three times with ethyl acetate (50 mL), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 30% ethyl acetate and petroleum ether) to obtain compound 17-1 (400 mg, yield: 68.7%) as a white solid. MS (ESI): m / z=254.1[M+H] + .

[0267] Compound 17 was obtained using a method similar to Example 4 using compound 17-1 as an intermediate.

[0268] 2-{[3-(1-amino-1-phenylethyl)-1-(2-methoxyethyl)pyrazolo[3,4-c]pyridin-5-yl]amino}-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0269] MS (ESI): m / z = 487.5 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.78(d,J=0.8Hz,1H),7.96(d,J=8.4Hz,1H),7.88(d,J=0.8Hz,1H),7.41-7.50(m,2H),7.35-7.33(m,2H),7.27- 7.25(m,1H),6.92(d,J=8.8Hz,1H),4.68(t,J=4.8Hz,2H),3.88(t,J=5.6Hz,2H),3.32(s,3H),2.92(s,2H),2.00(s,3H),1.50(s,6H).

[0270] Example 18: 2-{[3-(1-amino-1-cyclopropylethyl)-1-[2-(dimethylamino)ethyl]pyrazolo[3,4-c]pyridin-5-yl]amino}-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0271] Step 1: 1-{5-chloro-1-[2-(dimethylamino)ethyl]pyrazolo[3,4-c]pyridin-3-yl}ethan-1-one

[0272] Under argon, compound A1 (260 mg, 1.33 mmol), (2-bromomethyl)dimethylamine (222 mg, 1.46 mmol), and cesium carbonate (866 mg, 2.66 mmol) were mixed uniformly, and N,N-dimethylformamide (2.5 mL) was added. The mixture was reacted at room temperature under argon for 16 hours. The reaction was quenched with water (30 mL) and extracted three times with ethyl acetate (50 mL). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 0-100% ethyl acetate in petroleum ether, followed by a mixture of 1-5% methanol in dichloromethane) to obtain compound 18-1 (182 mg, yield: 38.0%) as a light yellow foamy solid. MS (ESI): m / z = 267.1 [M+H] + .

[0273] Compound 18 was obtained using a method similar to Example 4 using compound 18-1 as an intermediate.

[0274] 2-{[3-(1-amino-1-cyclopropylethyl)-1-[2-(dimethylamino)ethyl]pyrazolo[3,4-c]pyridin-5-yl]amino}-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0275] MS (ESI): m / z = 464.3 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.83(d,J=1.2Hz,1H),8.62(d,J=0.8Hz,1H),8.04(d,J=8.8Hz,1H),7.14(d,J=8.8Hz,1H),4.59(t,J=6 .4Hz,2H),3.12(s,2H),2.90(t,J=6.8Hz,2H),2.31(s,6H),1.65(s,3H),1.51(s,6H),1.35-1.44(m,1H),0.37-0.59(m,4H).

[0276] Example 19: 2-{[3-(1-amino-1-cyclobutylethyl)-1-methylpyrazolo[3,4-c]pyridin-5-yl]amino}-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0277] Compound 19 was obtained using a method similar to Example 6 using compound 6-0 as the starting material.

[0278] 2-{[3-(1-amino-1-cyclobutylethyl)-1-methylpyrazolo[3,4-c]pyridin-5-yl]amino}-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0279] MS (ESI): m / z = 421.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.77(dd,J=10.4,1.2Hz,1H),8.57(d,J=6.0Hz,1H),8.06-8.01(m,1H),7.30-7.02(m,1H),4.21-4.0 6(m,3H),3.14-3.09(m,2H),3.04-2.89(m,1H),1.99-1.73(m,4H),1.65(d,J=7.6Hz,3H),1.51(s,6H),1.35-1.26(m,2H).

[0280] Example 20 and Example 21: 2-({3-[(1S)-1-amino-1-cyclobutylethyl]-1-methylpyrazolo[3,4-c]pyridin-5-yl}amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one and 2-({3-[(1R)-1-amino-1-cyclobutylethyl]-1-methylpyrazolo[3,4-c]pyridin-5-yl}amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0281] Example 19 (100 mg, 0.24 mmol) was separated by SFC to give compound 20 (23 mg, yield: 22.6%) and compound 21 (23 mg, yield: 23.4%) as white solids. MS (ESI): m / z = 421.2 [M+H] + .

[0282] Example 22: 2-((3-(2-aminopent-4-en-2-yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0283] Step 1: 2-(5-chloro-1-methyl-1H-pyrazolo[3,4-c]pyridin-3-yl)pent-4-en-2-ol

[0284] Under argon, compound 1-4 (150 mg, 0.72 mmol) and anhydrous tetrahydrofuran (2 mL) were added to a three-necked reaction flask equipped with a magnetic stirrer. After the solution was cooled to -78°C, a solution of allylmagnesium bromide in tetrahydrofuran (1N, 0.72 mL, 0.72 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred at this temperature for 1 hour. Then, a solution of allylmagnesium bromide in tetrahydrofuran (1N, 0.20 mL, 0.20 mmol) was slowly added dropwise and the reaction continued for 1 hour. Water (10 mL) was added to quench the reaction, followed by extraction twice with ethyl acetate (40 mL). The combined organic phases were washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude compound 22-1 (200 mg, yield >100%) as a light yellow oil. The crude product was used directly in the next reaction. MS (ESI): m / z = 252.0 [M+H] + .

[0285] Compound 22-3 was obtained using compound 22-1 as an intermediate in a manner similar to Example 17 as a colorless oil. MS (ESI): m / z = 433.1 [M+H] + .

[0286] Step 4: 2-((3-(2-aminopent-4-en-2-yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0287] To a solution of compound 22-3 (30 mg, 0.069 mmol) in tetrahydrofuran (1 mL) and water (0.25 mL) was added triphenylphosphine (54 mg, 0.21 mmol). The mixture was heated to 70°C for 2 days, and the reaction solution was concentrated to obtain a crude product. The crude product was separated and purified by Prep-HPLC to obtain compound 22 (5 mg, yield: 15.6%) as a white solid. MS (ESI): m / z = 407.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.71(d,J=1.2Hz,1H),8.51(s,1H),7.97(d,J=9.2Hz,1H),7.08(d,J=8.8Hz,1H),5.56-5.52(m ,1H),5.04-4.95(m,2H),4.05(s,3H),3.04(s,2H),2.78-2.73(m,1H),2.66-2.60(m,1H),1.61(s,3H),1.43(s,6H).

[0288] Example 23: 2-((3-(2-aminopentan-2-yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0289] Compound 22-3 was used as an intermediate and a method similar to Example 17 was used to obtain compound 23 (25 mg, yield: 44.1%) as a white solid. MS (ESI): m / z = 409.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.78(s,1H),8.58(s,1H),8.05(d,J=8.8Hz,1H),7.15(d,J=8.8Hz,1H),4.13(s,3H),3.13( s,2H),2.05-1.94(m,2H),1.68(s,3H),1.52(s,6H),1.34-1.27(m,1H),1.16-1.11(m,1H),0.89(t,J=7.2Hz,3H).

[0290] Example 26: 2-{[3-(2-aminopropyl-2-yl)-1-methylpyrazolo[3,4-c]pyridin-5-yl]amino}-7,7,8-trimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one 3-amino-2H-[1,2'-bipyridine]-2-one

[0291] Step 1: 2-{[3-(2-azidoprop-2-yl)-1-methylpyrazolo[3,4-c]pyridin-5-yl]amino}-7,7,8-trimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0292] Under argon protection, compound 15-2 (80 mg, 0.32 mmol), 2-amino-7,7,8-trimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (73 mg, 0.35 mmol), Brettphos Pd G4 (30 mg, 0.032 mmol), Brettphos (17 mg, 0.032 mmol) and potassium phosphate (170 mg, 0.8 mmol) were mixed and then anhydrous 1,4-dioxane (2 mL) was added. The mixture was heated to 70 ° C under nitrogen protection and reacted for 12 hours. The reaction solution was cooled to room temperature, quenched with water (20 mL), extracted three times with ethyl acetate (50 mL), and the organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 30% ethyl acetate and petroleum ether) to obtain compound 26-1 (92 mg, yield: 68%) as a light yellow solid. MS (ESI): m / z = 421.3 [M+H] + .

[0293] Step 2: 2-{[3-(2-aminopropyl-2-yl)-1-methylpyrazolo[3,4-c]pyridin-5-yl]amino}-7,7,8-trimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0294] Compound 26-1 (92 mg, 0.22 mmol) and platinum dioxide (50 mg, 0.22 mmol) were mixed thoroughly, and ethyl acetate (5 mL) was added. The reaction mixture was allowed to react at room temperature under hydrogen for 1 hour. Water (20 mL) was added to quench the mixture, and the mixture was extracted three times with ethyl acetate (30 mL). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was dissolved in N,N-dimethylformamide (3 mL) and purified by Prep-HPLC (gradient elution with 20-90% acetonitrile in water containing 0.1% ammonium bicarbonate) to obtain compound 26 (30.7 mg, yield: 35.4%) as a white solid. MS (ESI): m / z = 395.3 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.88-8.59(m,2H),8.03(d,J=8.8Hz,1H),7.08(d,J=8.4Hz,1H),4.11(s,3H ),3.09-3.14(m,1H),1.70(s,3H),1.69(s,3H),1.52(s,3H),1.46(d,J=7.2Hz,3H),1.40(s,3H).

[0295] Example 27: 3-(1-amino-1-cyclopropylethyl)-N-(3-isopropyl-3H-imidazo[4,5-b]pyridin-5-yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-amine

[0296] Step 1: 5-Bromo-3-isopropyl-3H-imidazo[4,5-b]pyridine

[0297] To a solution of compound 27-0 (1.0 g, 5.05 mmol) in N,N-dimethylformamide (10 mL) at 0°C was added 60% sodium hydride (303 mg, 7.58 mmol) in three portions. The reaction mixture was stirred at room temperature for 30 minutes. After cooling to 0°C, 2-iodopropane (2.58 g, 15.2 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction was quenched with water (100 mL) and then extracted three times with ethyl acetate (100 mL). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 1-100% ethyl acetate in petroleum ether) to obtain compound 27-1 (618 mg, yield: 51.0%) as a yellow solid. MS (ESI): m / z = 240.0 [M+H] + .

[0298] Step 2: N-(2,4-dimethoxybenzyl)-3-isopropyl-3H-imidazo[4,5-b]pyridin-5-amine

[0299] Under argon protection, compound 27-1 (300 mg, 1.25 mmol), 2,4-dimethoxybenzylamine (627 mg, 3.75 mmol), Pd2(dba)3 (229 mg, 0.25 mmol), Xantphos (217 mg, 0.375 mmol), anhydrous cesium carbonate (814 mg, 2.50 mmol) and anhydrous N-methylpyrrolidone (6 mL) were added sequentially to a dry sealed tube equipped with a stirrer. The reaction mixture was replaced with argon three times and heated to 100°C for 2 hours. After the reaction was completed, the mixture was cooled to room temperature and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-10% methanol in dichloromethane) to obtain compound 27-2 (326 mg, yield: 79.9%) as a brown oil. MS (ESI): m / z = 327.1 [M+H] + .

[0300] Step 3: 3-Isopropyl-3H-imidazo[4,5-b]pyridin-5-amine

[0301] To a reaction solution of compound 27-2 (326 mg, 0.999 mmol) in N,N-dimethylformamide (5 mL) was added trifluoroacetic acid (5 mL). The reaction solution was stirred at room temperature for 30 minutes, concentrated, basified by adding methanolic ammonia (7 N), and concentrated to yield a crude product. The crude product was separated and purified by Prep-HPLC to yield compound 27-3 (125 mg, yield: 71.0%) as a yellow solid. MS (ESI): m / z = 177.1 [M+H] + .

[0302] Compound 27 was obtained using compound 27-3 as an intermediate using a method similar to that used to synthesize compound 1.

[0303] 3-(1-amino-1-cyclopropylethyl)-N-(3-isopropyl-3H-imidazo[4,5-b]pyridin-5-yl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-amine

[0304] MS (ESI): m / z = 391.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.74(s,1H),9.10(s,1H),8.80(s,1H),8.20(s,1H),7.84(d,J=8.7Hz,1H),7.00(d,J=8.7Hz,1H),4.98-4. 84(m,1H),4.04(s,3H),1.77(s,2H),1.60(s,3H),1.58(d,J=2.3Hz,6H),1.45-1.33(m,1H),0.49-0.35(m,2H),0.30-0.16(m,2H).

[0305] Example 28: 6-((3-(1-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-1-isopropyl-1,2-dihydro-3H-pyrazolyl[3,4-b]pyridin-3-one

[0306] Step 1: 6-chloro-1-isopropyl-1,2-dihydro-3H-pyrazolo[3,4-b]pyridin-3-one

[0307] To a solution of compound 28-0 (500 mg, 2.43 mmol) and isopropylhydrazine hydrochloride (537 mg, 4.85 mmol) in ethanol (5 mL) was added N,N-diisopropylethylamine (2.0 ml, 12.14 mmol) at room temperature. The reaction mixture was heated to 60°C and stirred for 2 hours. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse phase column (eluting with a gradient of 0-60% acetonitrile in water containing 0.1% ammonium bicarbonate) to obtain compound 28-1 (260 mg, yield: 50.6%) as a white solid. MS (ESI): m / z = 212.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),8.11(d,J=8.3Hz,1H),7.08(d,J=8.3Hz,1H),5.05-4.79(m,1H),1.39(d,J=6.7Hz,6H).

[0308] Step 2: 6-((2,4-dimethoxybenzyl)amino)-1-isopropyl-1,2-dihydro-3H-pyrazolo[3,4-b]pyridin-3-one

[0309] To a solution of compound 28-1 (230 mg, 1.09 mmol) and 2,4-dimethoxybenzylamine (909 mg, 5.43 mmol) in N-methylpyrrolidone (5 mL) at room temperature was added N,N-diisopropylethylamine (0.90 ml, 5.43 mmol). The reaction mixture was sealed, heated to 140°C, and stirred for 20 hours. After cooling, the reactant was directly separated and purified by reverse phase column chromatography to obtain compound 28-2 (250 mg, yield: 67.2%) as a yellow solid. MS (ESI): m / z = 343.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.28(s,1H),7.50(d,J=8.7Hz,1H),7.32-7.24(m,1H),7.22(d,J=8.3Hz,1H),6.56(d,J=2.4Hz,1H),6.46(dd ,J=8.3,2.4Hz,1H),6.25(d,J=8.7Hz,1H),4.75-4.65(m,1H),4.40(d,J=5.6Hz,2H),3.81(s,3H),3.73(s,3H),1.25(d,J=6.7Hz,6H).

[0310] Step 3: 6-amino-1-isopropyl-1,2-dihydro-3H-pyrazolo[3,4-b]pyridin-3-one

[0311] Using compound 28-2 as an intermediate, a method similar to that used to synthesize compound 27-3 was used to obtain crude compound 28-3 (156 mg) as a white solid. MS (ESI): m / z = 193.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.46(d,J=8.6Hz,1H),7.29-6.64(m,1H),6.34(s,2H),6.09(d,J=8.6Hz,1H),4.69-4.51(m,1H),1.17(d,J=6.7Hz,6H).

[0312] Compound 28 was obtained using a method similar to Example 1 using compound 28-3 as an intermediate.

[0313] 6-((3-(1-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-1-isopropyl-1,2-dihydro-3H-pyrazolyl[3,4-b]pyridin-3-one

[0314] MS (ESI): m / z = 407.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.71(s,1H),9.00(d,J=0.8Hz,1H),8.60(d,J=1.0Hz,1H),7.51(d,J=8.7Hz,1H),6.53(d,J =8.7Hz,1H),4.87-4.77(m,1H),3.84(s,3H),1.37(s,3H),1.19-1.11(m,7H),0.27-0.17(m,2H),0.12-0.03(m,2H).

[0315] Example 29: 7-((3-(1-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-1-methyl-2,3-dihydropyrido[2,3-d]pyrimidin-4(1H)-one

[0316] Step 1: 7-chloro-1-methylpyrido[2,3-d]pyrimidin-4(1H)-one

[0317] To a dry, sealed tube equipped with a stirrer, 2-amino-6-chloronicotinamide (630 mg, 3.16 mmol), triethyl orthoformate (2340 mg, 158 mmol), and concentrated hydrochloric acid (0.20 mL, 6.32 mmol) were added sequentially. The reaction mixture was purged with argon three times and heated to 60°C for 16 hours. The reaction mixture was cooled to room temperature, quenched with water (10 mL), and extracted with ethyl acetate (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluting with a mixture of 20% ethyl acetate in petroleum ether) to afford compound 29-1 (520 mg, yield: 61%) as a white solid. MS (ESI): m / z = 195.9 [M+H] + .

[0318] Step 2: 7-amino-1-methylpyrido[2,3-d]pyrimidin-4(1H)-one

[0319] Ammonia methanol solution (7N, 20 mL) was added to compound 29-1 (200 mg, 1.02 mmol) at room temperature, and the reaction mixture was heated to 80°C and stirred overnight. The mixture was cooled to room temperature and concentrated under reduced pressure to obtain a crude product. The crude product was slurried with a mixture of dichloromethane / methanol (8 / 1, 8 mL), filtered, and the filter cake was dried to obtain compound 29-2 (136 mg, yield: 75.5%) as a white solid. MS (ESI): m / z = 177.1 [M+H] + .

[0320] Compound 29 was obtained using a method similar to Example 1 using compound 29-2 as an intermediate.

[0321] 7-((3-(1-amino-1-cyclopropylethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-yl)amino)-1-methyl-2,3-dihydropyrido[2,3-d]pyrimidin-4(1H)-one

[0322] MS (ESI): m / z = 393.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.81(s,1H),8.91(d,J=0.9Hz,1H),8.82(d,J=1.1Hz,1H),7.74(s,1H),7.76-7.73(m,1H),6.48(d,J= 8.4Hz,1H),4.47(d,J=2.5Hz,2H),4.05(s,3H),3.09(s,3H),1.76(s,2H),1.51(s,3H),1.43-1.34(m,1H),0.38-0.18(m,4H).

[0323] Example 31: 2-(3-aminophenylmethyl)-1-methylpyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0324] Step 1: 5-chloropyrrolo[2,3-c]pyridin-3-yl)(phenyl)methanone

[0325] Under argon, benzoyl chloride (461 mg, 3.28 mmol) was added to a mixture of 5-chloropyrrolo[2,3-c]pyridine (500 mg, 3.28 mmol), aluminum trichloride (874 mg, 6.55 mmol), and dichloromethane (5 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction solution was quenched with sodium hydroxide solution (2N, 100 mL) and extracted three times with ethyl acetate (70 mL). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-30% ethyl acetate in petroleum ether) to obtain compound 31-1 (600 mg, yield: 71.3%) as a light yellow solid. MS (ESI): m / z = 257.0 [M+H] + .

[0326] Step 2: 5-chloro-1-methylpyrrolo[2,3-c]pyridin-3-yl)(phenyl)methanone

[0327] Under argon protection, iodomethane (713 mg, 5.03 mmol) was added to a mixture of 5-chloropyrrolo[2,3-c]pyridin-3-yl)(phenyl)methanone (430 mg, 1.68 mmol), cesium carbonate (655 mg, 2.01 mmol) and N,N-dimethylformamide (5 mL), and the reaction mixture was stirred at 0°C for 2 hours. The reaction was quenched with water (10 mL) and extracted three times with dichloromethane (25 mL). The combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-30% ethyl acetate in petroleum ether) to obtain compound 31-2 (400 mg, yield: 88.2%) as a yellow solid. MS (ESI): m / z = 271.0 [M+H] + .

[0328] Step 3: (5-chloro-1-methylpyrrolo[2,3-c]pyridin-3-yl)(phenyl)methylene)-2-methylpropane-2-sulfenamide

[0329] Under argon, compound 31-2 (300 mg, 1.11 mmol), 2-methylpropane-2-sulfenamide (134 mg, 1.11 mmol), tetraethyl titanate (1010 mg, 4.43 mmol), and anhydrous tetrahydrofuran (2 mL) were added sequentially to a dry, sealed tube equipped with a stirrer. The reaction mixture was purged with argon three times and heated to 90°C for 3 hours. After completion of the reaction, the mixture was cooled to room temperature, ethyl acetate (30 mL) and water (50 mL) were added, and stirred for 20 minutes. The organic phase was collected after filtration, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-30% ethyl acetate in petroleum ether) to obtain compound 31-3 (290 mg, yield: 70%) as a white solid. MS (ESI): m / z = 374.1 [M+H] + .

[0330] Step 4: N-(5-chloro-1-methylpyrrolo[2,3-c]pyridin-3-yl)(phenyl)methyl)-2-methylpropane-2-sulfenamide

[0331] Under argon, sodium borohydride (60 mg, 1.58 mmol) was added to a mixture of compound 31-3 (140 mg, 0.40 mmol) and tetrahydrofuran (1 mL). The reaction mixture was stirred at room temperature for 6 hours, quenched with water (5 mL), and extracted three times with dichloromethane (25 mL). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-30% ethyl acetate in petroleum ether) to obtain compound 31-4 (48 mg, yield: 34.1%) as a white solid. MS (ESI): m / z = 378.1 [M+H] + .

[0332] Step 5: N-(5-(7,7-dimethyl-5-oxo-7,8-dihydro-5H-pyrano[4,3-b]pyridin-2-yl)amino)-1-methyl-1H-pyrrolo[2,3-c]pyridin-3-yl)(phenyl)methyl)-2-methylpropane-2-sulfenamide

[0333] Under argon protection, compound 31-4 (48 mg, 0.14 mmol), 2-amino-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (26 mg, 0.14 mmol), Brettphos Pd G4 (12 mg, 0.013 mmol), Brettphos (9 mg, 0.02 mmol), cesium carbonate (110 mg, 0.34 mmol) and anhydrous 1,4-dioxane (0.5 mL) were added sequentially to a dry sealed tube equipped with a stirrer. The reaction mixture was replaced with argon three times and heated to 100 ° C for 2 hours. After the reaction was completed, it was cooled to room temperature and the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (gradient elution with a mixture of 0-30% ethyl acetate and petroleum ether) to obtain compound 31-5 (69 mg, yield: 99.8%) as a yellow solid. MS (ESI): m / z = 533.3 [M+H] + .

[0334] Step 6: 2-(3-aminophenylmethyl)-1-methylpyrrolo[2,3-c]pyridine-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0335] Under argon, a methanol solution of compound 31-5 (60 mg, 0.12 mmol) in 1 mL of methanol was added with a methanolic solution of hydrochloric acid (4 N, 0.4 mL, 1.6 mmol) and stirred at room temperature for 1 hour. After completion of the reaction, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by Prep-HPLC to obtain compound 31 (19 mg, yield: 37.9%) as a white solid. MS (ESI): m / z = 428.2 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ9.97(s,1H),8.51(s,1H),8.13(s,1H),7.90-7.88(d,J=8.8Hz,1H),7.48-7.46(m, 3H),7.30-7.26(m,2H),7.21-7.15(m,2H),5.28(s,1H),3.83(m,3H),3.08-2.96(m,2H),1.45-1.43(m,6H).

[0336] Example 32: 2-(3-(2-hydroxy-1-phenylpropyl-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0337] Compound 32-2 was obtained using compound 31-0 as an intermediate using a method similar to that used to synthesize compound 31-2.

[0338] Step 3: 2-(5-chloro-1-methylpyrrolo[2,3-c]pyridin-3-yl)-1-phenyl-2-propanol

[0339] Under argon, compound 32-2 (600 mg, 2.10 mmol) and anhydrous tetrahydrofuran (6 mL) were added to a three-necked reaction flask equipped with a magnetic stirrer. After the solution was cooled to -78°C, a solution of methyllithium in tetrahydrofuran (2N, 6.17 mL, 12.64 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred for 2 hours. After the reaction was complete, saturated ammonium chloride solution (20 mL) was added to quench the reaction, followed by extraction with ethyl acetate (10 mL) twice. The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 0-40% ethyl acetate in petroleum ether) to obtain compound 32-3 (420 mg, yield: 66.3%) as a yellow oil. MS (ESI): m / z = 301.0 [M+H] + .

[0340] Step 4: 2-(3-(2-hydroxy-1-phenylpropyl-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0341] Compound 32-3 was used as an intermediate and compound 32 was obtained by a method similar to that used to synthesize compound 1-7.

[0342] MS (ESI): m / z = 457.2 [M+H] + . 1 H NMR(400MHz,CD3OD)δ8.51(s,1H),8.25(s,1H),8.05-8.03(d,J=8.8Hz,1H),7.14-7.08(m,5H ),6.99-6.97(m,2H),3.84(s,3H),3.34-3.31(m,1H),3.11(s,2H),1.63(s,3H),1.50(s,6H).

[0343] Example 33 and Example 34:

[0344] Compound 32 (220 mg) was separated by SFC to give (R or S)-2-(3-(2-hydroxy-1-phenylpropyl-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridine-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (1st eluting isomer, 63 mg, yield: 28.6%) and (S or R)-2-(3-(2-hydroxy-1-phenylpropyl-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridine-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (2nd eluting isomer, 65 mg, yield: 29.5%) as white solids.

[0345] Example 33: (1 st eluting isomer)(R or S)-2-(3-(2-hydroxy-1-phenylpropyl-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0346] MS (ESI): m / z = 457.2 [M+H] +. 1 H NMR (400MHz, CD3OD) δ8.51 (s, 1H), 8.26 (s, 1H), 8.04-8.02 (d, J = 8.8Hz, 1H), 7.13-7.08 (m, 5H ),6.99-6.97(m,2H),3.84(s,3H),3.21-3.18(m,1H),3.11(s,2H),1.63(s,3H),1.50(s,6H).

[0347] Example 34: (2 nd eluting isomer)(S or R)-2-(3-(2-hydroxy-1-phenylpropyl-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0348] MS (ESI): m / z = 457.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.51 (s, 1H), 8.26 (s, 1H), 8.04-8.02 (d, J = 8.8Hz, 1H), 7.13-7.08 (m, 5H ),6.99-6.97(m,2H),3.84(s,3H),3.21-3.18(m,1H),3.11(s,2H),1.63(s,3H),1.50(s,6H).

[0349] Example 35: 2-(3-(2-amino-1-phenylpropyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-yl)amino)-7,8-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0350] Compound 32-3 was used as an intermediate and compound 35 was obtained by a method similar to Example 1.

[0351] MS (ESI): m / z = 456.2 [M+H] + . 1H NMR(400MHz,CD3OD)δ8.56(s,1H),8.44(s,1H),8.06-8.04(d,J=8.8Hz,1H),7.12-7.08(m,5H),6.85-6.83(m,2H),4.5 1-4.46(m,1H),3.83(s,3H),3.40-3.38(m,2H),3.20-3.14(m,1H),2.99-2.93(m,1H),1.68(s,3H),1.51-1.47(m,5H).

[0352] Examples 36-39:

[0353] Example 35 (38 mg) was separated by SFC to give (7S,8S)-2-(3-(S)-2-amino-1-phenylpropan-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,8-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (1 st eluting isomer, 6 mg, yield: 15.7%), and (7S,8S)-2-(3-(R)-2-amino-1-phenylpropan-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,8-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (2 nd eluting isomer, 5.4 mg, yield: 14.2%), (7R,8R)-2-(3-(S)-2-amino-1-phenylpropan-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,8-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (3 rd eluting isomer, 4.9 mg, yield: 12.8%), (7R,8R)-2-(3-(R)-2-amino-1-phenylpropan-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,8-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (4 th Eluting isomer, 5.7 mg, yield: 15%) were all white solids.

[0354] Example 36: (1 steluting isomer)(7S,8S)-2-(3-(S)-2-amino-1-phenylpropan-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,8-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0355] MS (ESI): m / z = 456.2 [M+H] + .

[0356] Example 37: (2 nd eluting isomer)(7S,8S)-2-(3-(R)-2-amino-1-phenylpropan-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,8-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0357] MS (ESI): m / z = 456.2 [M+H] + .

[0358] Example 38: (3 rd eluting isomer)(7R,8R)-2-(3-(S)-2-amino-1-phenylpropan-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,8-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0359] MS (ESI): m / z = 456.2 [M+H] + .

[0360] Example 39: (4 th eluting isomer)(7R,8R)-2-(3-(R)-2-amino-1-phenylpropan-2-yl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,8-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0361] MS (ESI): m / z = 456.2 [M+H] + .

[0362] Example 40 and Example 41:

[0363] Compound 40-4 was obtained using compound 31-0 as starting material in a similar manner to Example 32.

[0364] Compound 40-4 (220 mg, 0.48 mmol) was separated by SFC to give (R or S)-2-(3-(1-cyclopentyl-1-hydroxyethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridine-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (1st eluting isomer, 51 mg, yield: 39.2%), and (R or S)-2-(3-(1-cyclopentyl-1-hydroxyethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridine-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (2nd eluting isomer, 47 mg, yield: 36.2%), both as white solids.

[0365] Example 40: (1 st eluting isomer)(R or S)-2-(3-(1-cyclopentyl-1-hydroxyethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0366] MS (ESI): m / z = 435.0 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.49(s,1H),8.27(s,1H),8.03-8.01(d,J=8.8Hz,1H),7.33(s,1H),7.10-7.08(d,J=8.8Hz, 1H),3.89(s,3H),3.09(s,2H),2.61-2.57(m,1H),1.75-1.73(m,1H),1.65(s,3H),1.59-1.57(m,6H),1.51(s,6H).

[0367] Example 41: (2 nd Eluting isomer)(R or S)-2-(3-(1-cyclopentyl-1-hydroxyethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0368] MS (ESI): m / z = 435.0 [M+H] + . 1H NMR (400MHz, CD3OD) δ8.49(s,1H),8.27(s,1H),8.03-8.01(d,J=8.8Hz,1H),7.33(s,1H),7.10-7.08(d,J=8.8Hz, 1H),3.89(s,3H),3.09(s,2H),2.63-2.55(m,1H),1.76-1.72(m,1H),1.65(s,3H),1.59-1.53(m,6H),1.51(s,6H).

[0369] Example 42: 2-(3-(1-amino-1-cyclopentylethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0370] Step 1: (5-chloro-1-methylpyrrolo[2,3-c]pyridin-3-yl)(cyclopentyl)methylene)-2-methylpropane-2-sulfenamide

[0371] Compound 40-2 was used as an intermediate and a method similar to that used to synthesize compound 31-3 was used to obtain compound 42-1 as a yellow solid. MS (ESI): m / z = 366.1 [M+H] + .

[0372] Step 2: N-(5-chloro-1-methylpyrrolo[2,3-c]pyridin-3-yl)-1-cyclopentylethyl)-2-methylpropane-2-sulfenamide

[0373] Under argon, compound 42-1 (2.0 g, 5.47 mmol) and anhydrous tetrahydrofuran (15 mL) were added to a three-necked reaction flask equipped with a magnetic stirrer. After the solution was cooled to -78°C, a solution of methyllithium in tetrahydrofuran (1N, 480 mg, 21.9 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred for 2 hours. After the reaction was complete, saturated ammonium chloride solution (20 mL) was added to quench the reaction, followed by extraction with ethyl acetate (100 mL) twice. The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-100% ethyl acetate in petroleum ether) to obtain compound 42-2 (384 mg, yield: 18.4%) as a brown oil. MS (ESI): m / z = 382.1 [M+H] + .

[0374] Step 3: N-(1-cyclopentyl-1-(5-(7,7-dimethyl-5-oxo-7,8-dihydropyrano[4,3-b]pyridin-2-yl)amino)-1-methyl-1H-pyrrolo[2,3-c]pyridin-3-yl)ethyl)-2-methylpropane-2-sulfenamide

[0375] Under argon protection, compound 42-2 (300 mg, 0.79 mmol), 2-amino-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (151 mg, 0.79 mmol), Brettphos Pd G4 (72.3 mg, 0.079 mmol), Brettphos (55.6 mg, 0.12 mmol), anhydrous potassium phosphate (417 mg, 1.96 mmol) and anhydrous 1,4-dioxane (3 mL) were added sequentially to a dry sealed tube equipped with a stirrer. The reaction mixture was replaced with argon three times and heated to 70 ° C for 10 hours. After the reaction was completed, it was cooled to room temperature and the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (gradient elution with a mixture of 0-80% ethyl acetate in petroleum ether) to obtain the compound (240 mg, yield: 56.8%) as a yellow solid. MS (ESI): m / z = 538.3 [M+H] + .

[0376] Step 4: 2-(3-(1-amino-1-cyclopentylethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0377] Under argon, iodine (208 mg, 0.82 mmol) was added to a mixture of compound 42-3 (220 mg, 0.41 mmol) in tetrahydrofuran (1 mL) and water (1 mL). The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the mixture was filtered and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by Prep-HPLC to obtain compound 42 (4.3 mg, yield: 2.42%) as a white solid. MS (ESI): m / z = 434.2 [M+H] + . 1H NMR (400MHz, CD3OD) δ8.52 (s, 1H), 8.31 (s, 1H), 8.04-8.02 (d, J = 8.8Hz, 1H), 7.38 (s, 1H), 7. 12-7.10(d,J=8.8Hz,1H),3.90(s,3H),3.09(s,2H),2.73-2.69(m,1H),1.69-1.42(m,17H).

[0378] Example 43 and Example 44:

[0379] Compound 42 (124 mg, 0.29 mmol) was separated by SFC to give (R or S)-2-(3-(1-amino-1-cyclopentylethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (1 st eluting isomer, 29 mg, yield: 23.4%), and (R or S)-2-(3-(1-amino-1-cyclopentylethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (2 nd Eluting isomer, 27 mg, yield: 21.8%), all were white solids.

[0380] Example 43: (1 st eluting isomer)(R or S)-2-(3-(1-amino-1-cyclopentylethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0381] MS (ESI): m / z = 434.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.51 (s, 1H), 8.31 (s, 1H), 8.04-8.02 (d, J = 8.8Hz, 1H), 7.37 (s, 1H), 7. 12-7.10(d,J=8.8Hz,1H),3.90(s,3H),3.09(s,2H),2.72-2.67(m,1H),1.71-1.32(m,18H).

[0382] Example 44: (2 ndeluting isomer)(R or S)-2-(3-(1-amino-1-cyclopentylethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0383] MS (ESI): m / z = 434.2 [M+H] + .

[0384] Example 45: 2-(3-(1-amino-1-pyridin-3-yl)ethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0385] Step 1: 5-chloropyrrolo[2,3-c]pyridin-3-yl)(pyridin-3-yl)methanol

[0386] Under argon protection, 3-pyridinecarboxaldehyde (1.1 g, 10.3 mmol) was added to a mixture of compound 31-0 (1.5 g, 9.83 mmol) in methanol (16 mL) and water (2.5 mL). After stirring at 0°C for 10 minutes, sodium hydroxide (393 mg, 9.83 mmol) was added, and the reaction mixture was stirred at 0°C for 3 hours. After the reaction was complete, ethyl acetate (70 mL) was extracted three times, and the combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-35% ethyl acetate in petroleum ether) to obtain compound 45-1 (2.15 g, yield: 84.2%) as a white solid. MS (ESI): m / z = 259.9 [M+H] + .

[0387] Step 2: 5-chloropyrrolo[2,3-c]pyridin-3-yl)(pyridin-3-yl)methanone

[0388] Under argon, active manganese dioxide (4017 mg, 46.2 mmol) was added to a mixture of compound 45-1 (2000 mg, 7.7 mmol) and dichloromethane (35 mL), and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, water (30 mL) was added to quench the mixture, and the mixture was extracted three times with dichloromethane (70 mL). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-35% ethyl acetate in petroleum ether) to obtain compound 45-2 (340 mg, yield: 17.13%) as a pale white solid. MS (ESI): m / z = 257.9 [M+H] + .

[0389] 2-(3-(1-amino-1-pyridin-3-yl)ethyl)-1-methyl-1H-pyrrolo[2,3-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0390] Compound 45-2 was used as an intermediate and a method similar to Example 42 was used to obtain compound 45 as a white solid. MS (ESI): m / z = 443.1 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.70(s,1H),8.53(s,1H),8.45-8.43(d,J=8.8Hz,1H),7.97-7.92(m,2H),7.63-7.61(d,J= 8.8Hz,2H),7.42-7.38(m,1H),6.95-6.93(d,J=8.8Hz,1H),3.96(s,3H),2.93(s,2H),1.99(s,3H),1.51(s,6H).

[0391] Example 49: 2-(3-(2-aminopropyl-2-yl)-1-methylimidazo[1,5-a]pyrazin-6-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0392] Step 1: 1-(5-chloropyrazin-2-yl)ethylidene)-2-methylpropane-2-sulfenamide

[0393] Under argon protection, compound 49-0 (4.60 g, 29.38 mmol), S-tert-butylsulfenamide (7.12 g, 58.76 mmol), and tetrahydrofuran (70 mL) were added sequentially to a 250 mL sealed tube. Tetraethyl titanate (20.10 g, 88.14 mmol) was then added to the mixture. The sealed tube was heated and stirred in a 90°C oil bath for 4 hours. After the reaction solution cooled to room temperature, ethyl acetate (50 mL) was added to dilute the reaction solution. Saturated brine (50 mL) was then slowly added dropwise to the solution. After stirring for 10 minutes, the organic phase was collected, and the aqueous phase was extracted twice with ethyl acetate (20 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 0-30% ethyl acetate in petroleum ether) to obtain compound 49-1 (6.36 g, yield: 83.3%) as a light brown oil. MS (ESI): m / z = 186.0, 188.0 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.07 (s, 1H), 8.60 (d, J = 1.4Hz, 1H), 2.83 (s, 3H), 1.34 (s, 9H).

[0394] Step 2: 1-(5-chloropyrazin-2-yl)ethyl)-2-methylpropane-2-sulfenamide

[0395] Under argon, compound 49-1 (6.36 g, 24.49 mmol) and anhydrous tetrahydrofuran (90 mL) were added to a three-necked reaction flask equipped with a magnetic stirrer. After the solution was cooled to -78°C, a solution of lithium tri-sec-butylborohydride in tetrahydrofuran (1N, 49 mL, 49 mmol) was slowly added dropwise. After the addition was complete, the reaction mixture was stirred at -78°C for 2 hours. A solution of lithium tri-sec-butylborohydride in tetrahydrofuran (1N, 4.9 mL, 4.9 mmol) was further added to the reaction, and the reaction mixture was stirred for another hour. After the reaction was complete, saturated ammonium chloride (50 mL) was added to quench the reaction, followed by extraction with ethyl acetate (50 mL) three times. The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 0-40% ethyl acetate in petroleum ether) to obtain compound 49-2 (2.04 g, yield: 31.8%) as a light yellow oil. MS (ESI): m / z = 262.0, 264.0 [M+H] + .

[0396] Step 3: (S)-1-(5-chloropyrazin-2-yl)ethylamine-1-amine hydrochloride

[0397] To a solution of compound 49-2 (2.04 g, 7.79 mmol) in methanol (30 mL) was added a solution of hydrogen chloride / 1,4-dioxane (4N, 10 mL, 40 mmol) at room temperature, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction mixture was complete, the condensate was removed under reduced pressure to obtain a crude product. Water (30 mL) was added to the crude product, and the product was washed four times with ethyl acetate (20 mL). The combined aqueous phases were separated and purified by reverse-phase column chromatography to obtain compound 49-3 (1260 mg, yield: 83.2%) as a light brown solid. MS (ESI): m / z = 158.1, 160.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.87-8.86 (m, 4H), 8.75 (d, J = 1.2Hz, 1H), 4.65 (s, 1H), 1.55 (d, J = 6.8Hz, 3H).

[0398] Step 4: (S)-(1-(5-chloropyrazine-2-ethyl)amino)-2-methyl-1-oxopropyl-2-carbamic acid benzyl ester

[0399] To a mixture of compound 49-3 (519 mg, 2.68 mmol), 2-{[(benzyloxy)carbonyl]amino}-2-methylpropanoic acid (635 mg, 2.68 mmol), and anhydrous dichloromethane (10 mL) at room temperature was added N,N-diisopropylethylamine (1038 mg, 8.03 mmol). The solution was purged with nitrogen three times and stirred in an ice bath for 15 minutes. HATU (1527 mg, 4.02 mmol) was then added to the reaction solution. The ice bath was removed, and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-70% ethyl acetate in petroleum ether) to obtain compound 49-4 (1048 mg, yield: 103.9%) as a white solid. MS (ESI): m / z = 377.3, 379.3 [M+H] + .

[0400] Step 5: Benzyl 2-(6-chloro-1-methylimidazo[1,5-a]pyrazin-3-yl)propyl-2-carbamate

[0401] Under argon, Burgess reagent (948 mg, 3.98 mmol) was added to a solution of compound 49-4 (600 mg, 1.59 mmol) in anhydrous tetrahydrofuran (8 mL), and the reaction mixture was stirred at 70°C for 2 hours. After the reaction solution cooled to room temperature, ethyl acetate (25 mL) was added to dilute it, followed by washing with water (25 mL). The organic phase was collected, and the aqueous phase was extracted twice with ethyl acetate (15 mL). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-70% ethyl acetate in petroleum ether) to obtain compound 49-5 (143 mg, yield: 25.0%) as a light yellow solid. MS (ESI): m / z = 359.4, 361.4 [M+H] + .

[0402] Step 6: Benzyl 2-(6-((7,7-dimethyl-5-oxo-7,8-dihydro-5H-pyrano[4,3-b]pyridin-2-yl)amino)-1-methylimidazo[1,5-a]pyrazin-3-yl)propyl-2-carbamate

[0403] Under argon protection, compound 49-5 (120 mg, 0.33 mmol), 2-amino-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one (77 mg, 0.40 mmol), Brettphos Pd G4 (31 mg, 0.033 mmol), Brettphos (18 mg, 0.033 mmol), anhydrous potassium phosphate (142 mg, 0.669 mmol), anhydrous potassium acetate (65.6 mg, 0.70 mmol) and anhydrous 1,4-dioxane (2 mL) were added sequentially to a dry sealed tube equipped with a stirrer. The reaction mixture was replaced with argon three times and heated to 100 ° C. and stirred for 10 hours. After the reaction was completed, it was cooled to room temperature and the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 0-99% ethyl acetate in petroleum ether) to afford compound 49-6 (187 mg, yield: 86.9%) as a light brown solid. MS (ESI): m / z = 515.4 [M+H] + .

[0404] Step 7: 2-(3-(2-aminopropyl-2-yl)-1-methylimidazo[1,5-a]pyrazin-6-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0405] Under argon, triethylamine (30 mg, 0.29 mmol) and palladium acetate (6.6 mg, 0.029 mmol) were added to a solution of compound 49-6 (151 mg, 0.293 mmol) and triethylsilane (341 mg, 2.93 mmol) in anhydrous ethanol (2 mL). The mixture was purged with hydrogen three times and stirred at room temperature for 30 minutes. After conversion of most of the starting materials, the mixture was filtered and the filtrate was purified by Prep-HPLC to obtain compound 49 (28 mg, yield: 25.1%) as a light yellow solid. MS (ESI): m / z = 381.4 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ10.03(s,1H),9.93(s,1H),8.85(s,1H),7.93(d,J=8.8H z,1H),7.00(d,J=8.8Hz,1H),3.08(s,2H),2.47(s,3H),1.56(s,6H),1.41(s,6H).

[0406] Example 50: 2-(3-(2-aminopropyl-2-yl)-1-cyclopropylimidazo[1,5-a]pyrazin-6-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0407] Step 1: Benzyl 1-(((5-chloropyrazin-2-yl)methylamino)-2-methyl-1-oxopropan-2-ylcarbamate

[0408] To a mixture of compound 50-0 (700 mg, 2.95 mmol), 2-{[(benzyloxy)carbonyl]amino}-2-methylpropanoic acid (531 mg, 2.95 mmol), and anhydrous dichloromethane (10 mL) at 0°C was added N,N-diisopropylethylamine (953 mg, 7.35 mmol). The solution was purged with nitrogen three times and stirred in an ice bath for 15 minutes. HATU (1683 mg, 4.43 mmol) was then added to the reaction solution. The ice bath was removed, and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (gradient elution with a mixture of 0-70% ethyl acetate in petroleum ether) to obtain compound 50-1 (877 mg, yield: 81.9%) as a light yellow solid. MS (ESI): m / z = 363.2, 365.2 [M+H] + . 1H NMR (400MHz, CDCl3) δ8.45(s,1H),8.38(s,1H),7.34-7.32(m,5H),7.20(s,1H),5.22(s,1H),5.08(s,2H),4.56(s,2H),1.55(s,6H).

[0409] Step 2: Benzyl (2-(6-chloroimidazo[1,5-a]pyrazin-3-yl)propan-2-yl)carbamate

[0410] Under argon, Burgess reagent (1276 mg, 5.35 mmol) was added to a solution of compound 50-1 (777 mg, 2.14 mmol) in anhydrous tetrahydrofuran (12 mL). The reaction mixture was stirred at 50°C for 16 hours. After the reaction solution cooled to room temperature, ethyl acetate (25 mL) was added to dilute it, followed by washing with water (25 mL). The organic phase was collected, and the aqueous phase was extracted twice with ethyl acetate (15 mL). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by Prep-HPLC to obtain compound 50-2 (111 mg, yield: 14.3%) as a light white solid. MS (ESI): m / z = 345.1, 347.1 [M+H] + .

[0411] Step 3: Benzyl 2-(6-chloro-1-iodoimidazo[1,5-a]pyrazin-3-yl)propan-2-yl)carbamate

[0412] To a solution of compound 50-2 (185 mg, 0.537 mmol) in anhydrous dichloromethane (2 mL) at 0°C was added N-iodosuccinimide (133 mg, 0.59 mmol) in portions. The reaction mixture was stirred at 0°C for 1 hour, then allowed to warm to room temperature and stirred for 24 hours. After completion, the reaction was quenched with sodium sulfite solution (10%, 5 mL) and extracted three times with dichloromethane (10 mL). The combined organic phases were washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 0-50% ethyl acetate in petroleum ether) to obtain compound 50-3 (211 mg, yield: 83.5%) as a yellow solid. MS (ESI): m / z = 471.3, 473.3 [M+H] + . 1H NMR(400MHz, CDCl3)δ:8.54-8.52(m,1H),8.05(m,1H),7.29(m,3H),5.24-5.13(m,1H),4.98(s,2H),1.80(s,6H).

[0413] Step 4: (2-(6-chloro-1-cyclopropylimidazo[1,5-a]pyrazin-3-yl)propan-2-yl)benzylcarbamate

[0414] Under argon, compound 50-3 (110 mg, 0.234 mmol), cyclopropylboronic acid (30 mg, 0.35 mmol), Pd(dppf)Cl2·CH2Cl2 (19 mg, 0.023 mmol), anhydrous potassium phosphate (124 mg, 0.58 mmol), 1,4-dioxane (1.0 mL), and water (0.4 mL) were added sequentially to a dry, sealed tube equipped with a stirrer. The reaction mixture was purged with argon three times and allowed to react at 90°C for 2 hours. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (gradient elution with a mixture of 0-50% ethyl acetate in petroleum ether) to obtain compound 50-4 (72 mg, yield: 80.1%) as a light yellow solid. MS (ESI): m / z = 385.4, 387.4 [M+H] + .

[0415] 2-(3-(2-aminopropyl-2-yl)-1-cyclopropylimidazo[1,5-a]pyrazin-6-yl)amino)-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0416] Compound 50-4 was used as an intermediate and a method similar to Example 49 was used to obtain compound 50 as a light yellow solid. MS (ESI): m / z = 407.2 [M+H] + . 1 H NMR(400MHz, CDCl3)δ:9.54-9.50(m,1H),8.73(d,J=1.4Hz,1H),8.14(d,J=8.6Hz,1H),7.24-7.20 (m,1H),6.76-6.71(m,1H),3.08(s,2H),1.78-1.73(m,7H),1.30-1.23(m,5H),1.50-1.02(m,5H).

[0417] Example 51: 2-{[3-(2-aminopropan-2-yl)-1-(trifluoromethyl)imidazo[3,4-a]pyrazin-6-yl]amino}-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0418] Step 1: ({2-[6-chloro-1-(trifluoromethyl)imidazo[3,4-a]pyrazin-3-yl]propan-2-yl}amino)methanoic acid benzyl ester

[0419] Under argon, compound 50-3 (85 mg, 0.18 mmol), methyl 2,2-difluoro-2-(fluorodioxyidene-λ6-thio)acetate (69.4 mg, 0.36 mmol), and cuprous iodide (51.6 mg, 0.27 mmol) were mixed uniformly, and N-methylpyrrolidone (0.4 mL) was added. The mixture was heated to 110°C under argon and reacted for 1 hour. The reaction solution was cooled to room temperature, quenched with water (20 mL), and extracted three times with ethyl acetate (50 mL). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (eluted with a mixture of 30% ethyl acetate in petroleum ether) to obtain compound 51-1 (55 mg, yield: 74%) as a light yellow foamy solid. MS (ESI): m / z = 413.2 [M+H] + .

[0420] 2-{[3-(2-aminopropan-2-yl)-1-(trifluoromethyl)imidazo[3,4-a]pyrazin-6-yl]amino}-7,7-dimethyl-7,8-dihydro-5H-pyrano[4,3-b]pyridin-5-one

[0421] Compound 51-1 was used as an intermediate and a method similar to Example 49 was used to obtain compound 51 as a light yellow solid. MS (ESI): m / z = 435.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ10.19(d,J=1.2Hz,1H),8.97(s,1H),8.08(d,J=8.8Hz,1H),6.99(d,J=8.8Hz,1H),3.21(s,2H),1.75(s,6H),1.52(s,6H).

[0422] Example 52: 7-methyl-2-((1-methyl-3-(2-(methylamino)propan-2-yl)imidazo[1,5-a]pyrazin-6-yl)amino)-7,8-dihydro-5H-pyrazino[4,3-b]pyridin-5-one

[0423] Step 1: Benzyl (2-(6-chloro-1-methylimidazo[1,5-a]pyrazin-3-yl)propan-2-yl)(methyl)carbamate

[0424] Under argon protection and at 0°C, 60% sodium hydride (49 mg, 1.23 mmol) was added to a solution of compound 49-5 (220 mg, 0.61 mmol) in anhydrous tetrahydrofuran (6 mL). After stirring at 0°C for 20 minutes, iodomethane (522 mg, 3.68 mmol) was added dropwise. The reaction solution was stirred at room temperature for 20 hours. After the reaction was completed, the reaction solution was quenched with saturated aqueous ammonium chloride (5 mL), extracted twice with ethyl acetate (20 mL), and the organic phases were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse phase column chromatography to obtain compound 52-1 (81 mg, yield: 35.4%) as a yellow solid. MS (ESI): m / z = 373.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ8.79(d,J=1.1Hz,1H),7.71(s,1H),7.26-7.19(m,3 H),6.97-6.90(m,2H),4.78(s,2H),3.10(s,3H),2.41(s,3H),1.64(s,6H).

[0425] 7-Methyl-2-((1-methyl-3-(2-(methylamino)propan-2-yl)imidazo[1,5-a]pyrazin-6-yl)amino)-7,8-dihydro-5H-pyrazino[4,3-b]pyridin-5-one

[0426] Compound 52-1 was used as an intermediate and a method similar to Example 49 was used to obtain compound 52 as a yellow solid. MS (ESI): m / z = 364.2 [M-NHMe] + . 1H NMR (400MHz, DMSO-d6) δ10.05(s,1H),9.93(s,1H),8.86(d,J=1.3Hz,1H),7.93(d,J=8.7Hz,1H),7.00(d,J =8.7Hz,1H),3.08(s,2H),2.22-2.47(s,3H),2.14(m,1H),1.99(d,J=4.6Hz,3H),1.51(s,6H),1.42(s,6H).

[0427] Example 127: 2-(3-(1-amino-1-(3,3-difluorocyclobutyl)ethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydropyran-5H-pyrano[4,3-b]pyridin-5-one

[0428] Step 1: 1-(5-chloropyrazolo[3,4-c]pyridin-3-yl)-1-(3,3-difluorocyclobutyl)ethane-1-ol

[0429] Under argon, a solution of compound 1-1 (900 mg, 3.22 mmol) in anhydrous tetrahydrofuran (12 mL) was added to a three-necked reaction flask equipped with a magnetic stirrer. After the solution was cooled to -10°C, a solution of isopropylmagnesium chloride in tetrahydrofuran (2N, 4.0 mL, 8.00 mmol) was slowly added dropwise. After the addition was complete, the reaction mixture was stirred at -10°C for 10 minutes. Then, a solution of 1-(3,3-difluorocyclobutyl)ethan-1-one (864 mg, 6.44 mmol) in tetrahydrofuran (1.5 mL) was added dropwise to the reaction mixture, and stirring was continued for 1 hour. After the starting material was completely removed, the reaction was quenched by the addition of saturated ammonium chloride solution (20 mL). The mixture was then extracted three times with ethyl acetate (10 mL). The combined organic phases were washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by reverse phase column chromatography to obtain compound 127-1 (330 mg, yield: 35.6%) as a light yellow solid. MS (ESI): m / z = 287.9, 289.8 [M+H] + .

[0430] 2-(3-(1-amino-1-(3,3-difluorocyclobutyl)ethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydropyran-5H-pyrano[4,3-b]pyridin-5-one

[0431] Compound 127-1 was used as an intermediate, and the subsequent steps were similar to those in Example 6 to obtain compound 127 as a white solid. MS (ESI): m / z = 457.1 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.79(d,J=1.2Hz,1H),8.65(s,1H),8.05(d,J=8.8Hz,1H),7.12(d,J=8.8Hz,1H),4.13(s,3H),3.13( s,2H),2.90-2.79(m,1H),2.76-2.63(m,1H),2.62-2.46(m,2H),2.44-2.33(m,1H),1.65(s,3H),1.51(s,3H),1.50(s,3H).

[0432] Example 128 and Example 129:

[0433] Compound 127 (133 mg, 0.291 mmol) was separated by SFC to give (S or R)-2-(3-(1-amino-1-(3,3-difluorocyclobutyl)ethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydropyran-5H-pyrano[4,3-b]pyridin-5-one (1st eluting isomer, 29 mg, yield: 34.0%), and (R or S)-2-(3-(1-amino-1-(3,3-difluorocyclobutyl)ethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydropyran-5H-pyrano[4,3-b]pyridin-5-one (2nd eluting isomer, 29 mg, yield: 34.0%). isomer, 32 mg, yield: 42.9%), all were white solids.

[0434] Example 128: (1 st eluting isomer)(S or R)-2-(3-(1-amino-1-(3,3-difluorocyclobutyl)ethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydropyran-5H-pyrano[4,3-b]pyridin-5-one

[0435] MS (ESI): m / z = 457.1 [M+H] + . 1H NMR (400MHz, CD3OD) δ8.80(d,J=1.2Hz,1H),8.66(d,J=0.8Hz,1H),8.06(d,J=8.8Hz,1H),7.13(d,J=8.8Hz,1H),4.14(s,3H),3 .14(s,2H),2.90-2.81(m,1H),2.75-2.65(m,1H),2.63-2.47(m,2H),2.43-2.33(m,1H),1.66(s,3H),1.52(s,3H),1.51(s,3H).

[0436] Example 129: (2 nd eluting isomer)(R or S)-2-(3-(1-amino-1-(3,3-difluorocyclobutyl)ethyl)-1-methyl-1H-pyrazolo[3,4-c]pyridin-5-amino)-7,7-dimethyl-7,8-dihydropyran-5H-pyrano[4,3-b]pyridin-5-one

[0437] MS (ESI): m / z = 457.1 [M+H] + . 1 H NMR (400MHz, CD3OD) δ8.77(d,J=1.2Hz,1H),8.63(s,1H),8.03(d,J=8.8Hz,1H),7.10(d,J=8.8Hz,1H),4.12(s,3H),3.11( s,2H),2.88-2.78(m,1H),2.73-2.61(m,1H),2.59-2.46(m,2H),2.43-2.31(m,1H),1.64(s,3H),1.50(s,3H),1.49(s,3H).

[0438] A similar method to Example 1 was used to replace the corresponding starting materials to obtain the compounds shown in the following table:

[0439] A similar method to Example 31 was used to replace the corresponding starting materials to obtain the compounds shown in the following table:

[0440] A similar method to Example 49 was used to replace the corresponding starting materials to obtain the compounds shown in the following table:

[0441] HPK1 ADP-Glo ​​enzyme activity assay

[0442] Prepare enzyme activity assay buffer containing 40 mM Tris, pH 7.5; 20 mM MgCl2; 0.1 mg / mL BSA; and 50 μM DTT. Dissolve the compound in 100% DMSO to a 10 mM stock concentration. Serially dilute the compound in DMSO, starting at 100 μM, in triplicate for a total of eleven concentrations. Dilute the compound 1:20 in enzyme activity assay buffer and add 1 μL of the diluted compound to the working wells, with duplicates for each concentration. Add 1 μL of the 1:20 diluted DMSO solution to the negative and positive control wells. Prepare a 2.5× substrate / ATP working solution containing 0.25 μg / μL MBP protein and 45 μM ATP in enzyme activity assay buffer. Add 2 μL of the 2.5× substrate / ATP working solution to each working well. Prepare a 2.5× enzyme reaction working solution containing 0.5 ng / μL HPK1 recombinant protein (Signalchem, Catalog No. M23-11G-10) in enzyme activity assay buffer. Add 2 μL of 2.5× enzyme reaction working solution to each working well. Add only 2 μL of enzyme activity assay buffer to the negative control well. After sealing the plate with film, centrifuge briefly and incubate at room temperature for 30 minutes. After the reaction is complete, add 5 μL of ADP-Glo ​​reagent (Promega, Catalog No. V1901) to each well and incubate at room temperature for 40 minutes. Subsequently, add 10 μL of kinase assay reagent (Promega, Catalog No. V1901) and incubate at room temperature for 20 minutes. The final luminescence signal is measured.

[0443] Calculate the average value of positive wells and negative wells respectively as the positive control value (Signal pos ) and negative control values ​​(Signal neg ). Set the working hole signal value (Signal test ) According to the formula Inhibition rate = (Signal pos -Signal test ) / (Signal pos–Signal neg ) × 100% to calculate the inhibition rate. The obtained inhibition rate was used to draw the concentration-inhibition rate curve by nonlinear fitting in GraphPad Prism software to calculate IC 50 The experimental results are shown in Table 1.

[0444] Biological Test Example 2 GLK ADP-Glo ​​Enzyme Activity Test

[0445] Prepare enzyme activity assay buffer containing 40 mM Tris, pH 7.5; 20 mM MgCl2; 0.1 mg / mL BSA; and 50 μM DTT. Dissolve the compound in 100% DMSO to a 10 mM stock concentration. Serially dilute the compound in DMSO, starting at 1 mM, in three-fold increments for a total of eleven concentrations. Dilute the compound 1:20 in enzyme activity assay buffer and add 1 μL of the diluted compound to the working wells, with duplicates for each concentration. Add 1 μL of the 1:20 diluted DMSO solution to the negative and positive control wells. Prepare a 2.5× substrate / ATP working solution containing 0.5 μg / μL PKA substrate peptide and 105 μM ATP in enzyme activity assay buffer. Add 2 μL of the 2.5× substrate / ATP working solution to each working well. Prepare a 2.5× enzyme reaction working solution containing 2.5 ng / μL GLK recombinant protein (Signalchem, Catalog No. M25-11G-10) in enzyme activity assay buffer. Add 2 μL of 2.5× enzyme reaction working solution to each working well. Add only 2 μL of enzyme activity assay buffer to the negative control well. Apply the sealing film and briefly centrifuge. Incubate at room temperature for 1 hour. After the reaction is complete, add 5 μL of ADP-Glo ​​reagent (Promega, Catalog No. V1901) to each well and incubate at room temperature for 40 minutes. Subsequently, add 10 μL of kinase assay reagent (Promega, Catalog No. V1901). Incubate at room temperature for 20 minutes, and then measure the final luminescence signal.

[0446] Calculate the average value of positive wells and negative wells respectively as the positive control value (Signal pos ) and negative control values ​​(Signal neg ). Set the working hole signal value (Signal test ) According to the formula Inhibition rate = (Signal pos -Signal test ) / (Signal pos –Signal neg ) × 100% to calculate the inhibition rate. The obtained inhibition rate was used to draw the concentration-inhibition rate curve by nonlinear fitting in GraphPad Prism software to calculate IC 50 The experimental results are shown in Table 1.

[0447] Table 1 HPK1 and GLK enzyme activity test results

[0448] Where A represents IC 50 Value ≤ 1nM; B means 1nM<IC 50 Value ≤ 10nM; C means 10nM < IC 50 Value ≤ 100nM; D means 100nM<IC 50 value.

[0449] Biological Test Example 3 SLP76 Phosphorylation Cytological Test

[0450] Inhibition of HPK1 can inhibit the phosphorylation of its downstream protein SLP76. The phosphorylation of SLP76 protein was detected using Jurkat (ATCC, Clone E6-1 TIB-152 TM ) cells were tested. On the first day of the experiment, the cells were diluted to 10 6 / mL, 100μL per well, 10 5 Cells were plated in 96-well cell culture plates and starved for 4 hours. The compound was dissolved in 100% DMSO to a stock concentration of 4 mM. The compound DMSO solution was serially diluted fourfold starting from 4 mM, for a total of 9 concentrations. 4 μL of the diluted compound was diluted into 196 μL of 37°C preheated RPMI 1640 and mixed. 50 μL of the final diluted compound was added to the cells and incubated at 37°C for 20 minutes. 50 μL of the diluted human CD3 / CD28 T cell activator (Stemcell, Cat. No. 10971) was added to give a final concentration of 1 / 40 of the total volume of the activator, and the cells were incubated at 37°C for 30 minutes. After the reaction, the cells were centrifuged at 1200 rpm at 4°C for 5 minutes, the culture medium was aspirated, 150 μL of cell lysis buffer (provided in the ELISA kit, Cell Signaling, Cat. No. 30794C) was added and placed on ice for 30 minutes to fully lyse the cells. After the lysed cells were evenly pipetted, they were centrifuged at 4000 rpm at 4°C for 5 minutes, and then 50 μL of the supernatant was added to the FastScan TM Phospho-SLP-76 (Ser376) ELISA Kit (Cell Signaling, Cat. No. 30794C) was used to detect the phosphorylation level of cellular SLP76.

[0451] Calculate the average value of positive wells and negative wells respectively as the positive control value (Signal pos ) and negative control values ​​(Signal neg). Set the working hole signal value (Signal test ) According to the formula Inhibition rate = (Signal pos -Signal test ) / (Signal pos –Signal neg ) × 100% to calculate the inhibition rate. The obtained inhibition rate was used to draw the concentration-inhibition rate curve by nonlinear fitting in GraphPad Prism software to calculate IC 50 The experimental results are shown in Table 2.

[0452] Table 2 Results of SLP76 phosphorylation inhibition experiment

[0453] Where A represents IC 50 Value ≤ 100nM; B means 100nM < IC 50 Value ≤ 500nM; C means 500nM < IC 50 Value ≤1000nM.

[0454] Biological Test Example 4 CYP Enzyme Inhibition Test

[0455] Commercially available mixed human hepatocyte microsomes were used to evaluate the extent to which the compounds of the present invention inhibited the production of metabolites of each probe drug by measuring the amount of metabolites by LC / MS / MS, using acetaminophen (CYP 1A2), hydroxybupropion (CYP 2B6), N-desethylamodiaquine (CYP 2C8), 4-hydroxydiclofenac (CYP 2C9), 4-hydroxymephenytoin (CYP 2C19), dextrorphan (CYP 2D6), and 6β-hydroxytestosterone (CYP 3A4), which are typical substrate metabolic reactions of the major human CYP 7 molecular species (CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4).

[0456] The reaction solution consisted of the substrates phenazone (CYP1A2), bupropion (CYP2B6), amodiaquine (CYP2C8), diclofenac sodium (CYP2C9), s-mephenytoin (CYP2C19), dextromethorphan (CYP2D6), and testosterone (CYP3A4), along with 0.2 mg / mL of mixed human liver microsomes. 0.1 M PBS buffer was added, along with nicotinamide adenine dinucleoside phosphate as a coenzyme to initiate the reaction. The metabolites were quantified using LC / MS / MS. The compounds of the invention were tested at concentrations of 50, 15, 5, 1.5, 0.5, 0.15, 0.05, 0.015, and 0 μM (eight non-zero concentration points).

[0457] The residual activity (%) of the compounds of the present invention at various concentrations was calculated using the following formula: % Inhibition = 100 / (1 + 10^((LogIC50 - X) * HillSlope)). Sigmoidal dose-inhibition curves were plotted using a nonlinear regression model and IC50 values ​​were calculated using Graph Pad Prism 9.0 software. The experimental results are shown in Table 3.

[0458] Table 3 CYP enzyme inhibition test results of some compounds

[0459] The results showed that the inhibitory effect of the compounds of the present application on CYP enzymes was weak, thus avoiding drug-drug interactions with other drugs.

[0460] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound represented by the following formula I or formula II, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof; in, X 1 Selected from the following group: CR 3 NR 3 ; X 2 Selected from the group consisting of: N, CH; X 4 N or CH; Z 1 is N or C; and It is an aromatic ring; R 3 Selected from the following group: H, C 1-6 Alkyl, C 3-8 Cycloalkyl (preferably C 3-6 cycloalkyl), a 4-8 membered heterocyclic group containing 1-3 heteroatoms selected from N, O, and S, a 4-8 membered (preferably 5 membered) heteroaryl group containing 1-3 heteroatoms selected from N, O, and S, and the above groups may be further substituted by 1-3 groups selected from the following groups: D, -NR 11 R 12 、-CN、halogen、C 1-6 Alkyl, C 1-6 Alkoxy, -C(O)R 11 、-SO2R 18 ; R 11 , R 12 Each independently selected from H, C 1-3 Alkyl, or R 11 、R 12 Together with the atoms to which they are attached, they form a 4-6 membered heterocyclic ring; R 18 C 1-3 alkyl; R 1 for: Among them, R a and R b Each independently selected from the group consisting of: H, C 1-6 Alkyl, phenyl, 5-7 membered heteroaryl, C 3-8 Cycloalkyl, C 1-6 Alkenyl, benzyl, C 5-8 Spiroalkyl and C 5-8 Bridged cycloalkyl, 4-12 membered (preferably 4-8 membered) heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O, 5-7 membered heteroaromatic substituted C 1-6 Alkyl, or R a and R b The atoms connected to it together form C 3-8 Cycloalkyl; And the R a or R b , or R a and R b The ring formed by the atoms connected thereto may be further substituted by 1-3 groups selected from the group consisting of halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl; R c Selected from the group consisting of: -NR 16 R 17 ,-OH;R 16 , R 17 Each independently selected from H, C 1-3 alkyl; R 2 It has the structure shown below: Where n is 0 or 1; Y 1 N or CH; Each Y 4 and Y 5 Each independently selected from the following group: O, NR 15 、C(R 13 R 14 ); each R 13 , R 14 , R 15 Each independently selected from the following group: H, C 1-6 alkyl; R 4 、R 5 Each independently selected from the group consisting of: H, C 1-6 Alkyl (preferably C 1-3 alkyl); In the compound of formula I, each chiral center is in R configuration or S configuration.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein: The compound of formula I has the structure shown in the following formula I-1 and I-2: Among them, X 1 Selected from the following group: NR 3 , CR 3 ; X 2 Selected from the group consisting of: N, CH; X 4 N or CH; R 1 、R 2 The definition is as stated in claim 1.

3. The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein: The R a and R b Each independently selected from the group consisting of: H, C 1-6 Alkyl, phenyl, 5-7 membered heteroaryl, C 3-8 Cycloalkyl, C 1-6 alkenyl, benzyl, a 4-8 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O; or R a and R b The atoms connected to it together form C 3-8 Cycloalkyl; and said R a or R b It may be further substituted by 1-3 groups selected from the group consisting of halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 Cycloalkyl.

4. The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein: R 3 Select from the following group: C 1-6 Alkyl, C 3-8 Cycloalkyl (preferably C 3-6 cycloalkyl), a 4-8 membered heterocyclic group containing 1-3 heteroatoms selected from N, O, and S, a 4-8 membered (preferably 5 membered) heteroaryl group containing 1-3 heteroatoms selected from N, O, and S, and the above groups may be further substituted by 1-3 groups selected from the following groups: D, -NR 11 R 12 , halogen, C 1-6 Alkyl, C 1-6 alkoxy; R 11 , R 12 Selected from H, C 1-3 Alkyl, or R 11 、R 12 Together with the atoms they are connected to, they form a 4-6 membered heterocyclic ring.

5. The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein: The compound of formula I has a structure selected from the group consisting of:

6. The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein: The R2 is selected from the following group: Among them, the R 4 、R 5 、R 13 、R 14 、R 15 The definition is as stated in claim 1.

7. The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein: The compound of formula I has the following structure: Among them, the X 1 、X 2 、R 4 、R 5 、R a 、R b 、R c 、Y 4 、Y 5 , n is defined as in claim 1.

8. The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein: The compound of formula I has the following structure: Among them, the X 2 、R 3 、R 4 、R 5 、R a 、R b 、R c 、Y 5 The definition is as stated in claim 1.

9. The compound according to claim 1, or a pharmaceutically acceptable salt, optical isomer or hydrate thereof, wherein: The compound of formula I is selected from the following group:

10. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises (1) the compound according to claim 1 or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate; and (2) a pharmaceutically acceptable carrier.

11. Use of the compound according to claim 1 or its stereoisomer or tautomer, or its pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition according to claim 9, characterized in that: Used for preparing a pharmaceutical composition for preventing and / or treating diseases related to the activity or expression of HPK1 kinase.

12. A combination chemotherapy regimen for a compound according to any one of claims 1 to 9, and pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates, and deuterated compounds thereof, with other tumor immunotherapeutic agents, wherein the other tumor immunotherapeutic agents are selected from the group consisting of small molecule compounds and antibodies (including but not limited to PD-1, PD-L1, CTLA-4, TIM-3, TGF-β and its receptors, LAG3 antagonists, or TLR4, TLR7, TLR8, TLR9, STING agonists, etc.), radiotherapy regimens, tumor-targeted drugs, and tumor vaccines.

13. Use of the compound according to any one of claims 1 to 9 and its pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, solvates and deuterated compounds in combination with CAR-T immunotherapy in cancer immunotherapy.

14. The use according to claim 13, characterized in that The cancer is selected from the group consisting of lymphoma, blastoma, medulloblastoma, retinoblastoma, sarcoma, liposarcoma, synovial cell sarcoma, neuroendocrine tumor, carcinoid tumor, gastrinoma, islet cell carcinoma, mesothelioma, schwannoma, acoustic neuroma, meningioma, adenocarcinoma, melanoma, leukemia or lymphoid malignancy, squamous cell carcinoma, epithelial squamous cell carcinoma, lung cancer, small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous cell carcinoma of the lung, peritoneal cancer, hepatocellular carcinoma, gastric cancer, intestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, Liver cancer, breast cancer, metastatic breast cancer, colon cancer, rectal cancer, colorectal cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, hepatocellular carcinoma, anal cancer, penile cancer, Merkel cell carcinoma, esophageal cancer, biliary tract tumors, head and neck cancer, and hematological malignancies.

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