A class of pyridazinone heterocyclic compounds, their preparation methods and uses

Abstract

This invention relates to a pyridazinone heterocyclic compound, its preparation, and its use. Specifically, this invention relates to compounds of formula I or pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites, or prodrugs thereof, pharmaceutical compositions and kits comprising the same, methods for their preparation, and their use in the preparation of medicaments for the prevention or treatment of Axl-related diseases (particularly neoplastic diseases).

Description

Technical Field

[0001] This invention belongs to the field of medicine, specifically relating to a compound with Axl inhibitory activity, its preparation and uses. Background Technology

[0002] Currently, 58 receptor tyrosine kinases (RTKs) have been discovered in humans. These are transmembrane receptors on the cell surface, mainly composed of an extracellular domain, a transmembrane domain, and an intracellular domain, among which the intracellular domain possesses kinase activity. After binding to a ligand, RTKs undergo dimerization and autophosphorylation, thereby activating themselves and subsequently activating downstream signaling cascades within the cell, regulating functions such as mitosis, cell cycle, cell adhesion, migration, and angiogenesis.

[0003] The TAM family belongs to the receptor tyrosine kinase subfamily, and its members include Tyro-3, Axl, and Mer. Among them, Axl and Mer regulate a variety of cellular functions in signal transduction pathways. Axl (Ufo, Ark, or Tyro7) was discovered in the DNA of human chronic myeloid leukemia by Bryan et al. in 1991. Axl not only has a similar gene structure to Tyro3 and a similar tyrosine kinase domain amino acid sequence to Mer, but they also share a common growth arrest-specific protein 6 (Gas6) ligand.

[0004] Axl activation occurs in several different ways: (1) homodimerization through ligand binding; (2) homodimerization independent of ligand binding, i.e., self-dimerization due to Axl overexpression; (3) heterodimerization of Axl receptor with two other TAM receptors; (4) heterodimerization of Axl receptor with non-TAM receptors; and (5) activation by binding to the extracellular domains of two different cells. Ligand binding is the primary mechanism of Axl activation. After Gas6 binds to the extracellular domain of Axl, Axl dimerizes, leading to phosphorylation of three phosphorylation sites (Y779, Y821, and Y866) in the intracellular domain. These phosphorylation sites can bind to phosphatidylinositol 3-kinase subunit (PI3K), phospholipase C (PLC), and growth factor receptor-binding protein 2 (Grb2), activating multiple related signaling pathways such as RAS / ERK and PI3K / Akt, promoting cellular inflammation, cell survival, migration, invasion, and metastasis. Other activation mechanisms require overexpression of the Axl receptor and other Axl-binding receptors.

[0005] Besides being highly expressed in various cells and tissues in the human body, Axl has also been found to be highly expressed in a variety of malignant tumors, including lung cancer, breast cancer, colon cancer, gastric cancer, liver cancer, and ovarian cancer. Abnormal Axl expression can antagonize tumor cell apoptosis, promote tumor cell invasion and metastasis, promote tumor angiogenesis, and drive tumor occurrence and development, closely related to tumor development, recurrence, and poor prognosis. Recent studies have shown that high Axl expression may mediate acquired resistance to EGFR; clinical studies indicate that approximately 20% of EGFR-resistant patients have high Axl expression. Preclinical studies have found that the combination of Axl inhibitors and EGFR inhibitors can effectively overcome and improve EGFR inhibitor resistance, and clinical application of this combination therapy is underway. Furthermore, Axl is highly expressed in the tumor microenvironment (such as macrophages and dendritic cells), and can synergistically promote tumor progression through interactions with tumor cells and other stromal cells. Abnormal activation caused by Axl overexpression is also closely related to resistance to other targeted drugs and chemotherapy drugs.

[0006] There remain significant unmet clinical needs in the treatment of human cancer. Based on the proven anti-tumor effects of Axl inhibitors in preclinical studies and the large number of ongoing clinical research projects, the development of Axl inhibitors has become a hot topic in anti-tumor drug research. In conclusion, there is an urgent need in this field to develop novel small-molecule Axl inhibitors. Summary of the Invention

[0007] The first aspect of the invention relates to compounds of formula I or pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites, or prodrugs thereof.

[0008]

[0009] in,

[0010] R 1 Selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic groups, C6-C 10 aryl and 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic, C6-C 10 aryl or 5-10 heteroaryl groups are optionally surrounded by one or more R groups. 5 replace;

[0011] R 5 Each occurrence is independently selected from halogens, -CN, -OH, -NH2, -NO2, and -S(O). p R A -COR A=O, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 alkoxy, 3-8 membered heterocyclic, C2-C6 alkenyl, C2-C6 alkynyl, C6-C 10 aryl and 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 alkoxy, 3-8 membered heterocyclic, C2-C6 alkenyl, C2-C6 alkynyl, C6-C 10 aryl or 5-10 heteroaryl groups are optionally surrounded by one or more R groups. 6 replace;

[0012] R 6 Each occurrence is independently selected from halogens, -CN, -OH, -NH2, -NO2, and -S(O). p R A -COR A S = O;

[0013] R A Independently selected from H and C1-C6 alkyl groups;

[0014] p is selected from 1 and 2;

[0015] R 2 Each of the elements, when appearing independently, is selected from H, -OH, halogen, -CN, -NH2, -NO2, C1-C6 alkyl, C1-C6 alkoxy, and C3-C8 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, or C3-C8 cycloalkyl is optionally separated by one or more R. 7 replace;

[0016] R 7 Each of these elements is independently selected from -OH, halogen, -CN, -NH2, -NO2, C1-C6 alkoxy, C1-C6 alkyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy in each occurrence.

[0017] m is selected from 0, 1, 2, 3, and 4;

[0018] R 3 Selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic groups, C6-C 10 aryl and 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic, C6-C 10 aryl or 5-10 heteroaryl groups are optionally surrounded by one or more R groups. 8 replace;

[0019] R 8Each of these elements is independently selected from -OH, halogen, -CN, -NH2, -NO2, C1-C6 alkoxy, C1-C6 alkyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy in each occurrence.

[0020] R 4 Each of the following groups, when appearing independently, is selected from H, -OH, halogen, CN, -NH2, -NO2, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, and 3-8 membered heterocyclic groups, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, or 3-8 membered heterocyclic group is optionally replaced by one or more R groups. 9 replace;

[0021] R 9 Each of these groups is independently selected from -OH, halogen, -CN, -NH2, -NO2, C1-C6 alkoxy, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy, and 3-8 membered heterocyclic groups when it appears.

[0022] n is selected from 0, 1, and 2; and

[0023] X is selected from CH and N.

[0024] On the other hand, the present invention provides a pharmaceutical composition comprising a preventive or therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug, and one or more pharmaceutically acceptable carriers.

[0025] On the other hand, the present invention provides a medicine box comprising:

[0026] a) The compounds of the present invention or their pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites, or prodrugs, or the pharmaceutical compositions of the present invention; and

[0027] b) Optional packaging and / or instructions.

[0028] On the other hand, the present invention provides compounds of the present invention or pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs thereof, or pharmaceutical compositions of the present invention, or kits of the present invention, for the prevention or treatment of diseases (particularly tumor diseases) associated with Axl.

[0029] On the other hand, the present invention provides the use of the compounds of the present invention or pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs of the present invention, or pharmaceutical compositions of the present invention, or cassettes of the present invention in the preparation of medicaments for the prevention or treatment of diseases (particularly tumor diseases) associated with Axl.

[0030] On the other hand, the present invention provides a method for preventing or treating diseases (particularly tumor diseases) associated with Axl, comprising administering to an individual in need a preventive or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug of the present invention, or a pharmaceutical composition of the present invention, or a cassette of the present invention.

[0031] On the other hand, the present invention provides a method for preparing the compounds of the present invention, comprising the steps shown in reaction route 1 or reaction route 8 below:

[0032] Reaction route 1

[0033]

[0034] Reaction route 8

[0035]

[0036] Wherein, LG represents a leaving group, which includes, but is not limited to, halogen atoms, methanesulfonyloxy groups, trifluoromethanesulfonates, etc.; R 1 R 2 R 3 R 4 X, m, and n are as defined above. Detailed Implementation

[0037] Before further describing the invention, it should be understood that the invention is not limited to the specific embodiments described herein. It should also be understood that the terminology used herein is for descriptive purposes only and not for limiting the specific embodiments.

[0038] definition

[0039] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to technical terms herein refer to techniques commonly understood in the art, including variations or equivalent substitutions of techniques that are obvious to one of ordinary skill in the art. While it is believed that the following terms will be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain the invention.

[0040] As used herein, the terms “including,” “comprising,” “having,” “containing,” or “involving,” and their other variations herein, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

[0041] As used herein, the term "alkyl" is defined as a straight-chain or branched saturated aliphatic hydrocarbon group. For example, as used herein, the term "C1-C6 alkyl" refers to a straight-chain or branched group having 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, or n-hexyl), optionally substituted with one or more (such as 1 to 3) suitable substituents such as halogens.

[0042] As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated nonaromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or bicyclic, including spirocyclic, fused or bridged systems, such as bicyclic [1.1.1]pentyl, bicyclic [2.2.1]heptyl, bicyclic [3.2.1]octyl or bicyclic [5.2.0]nonyl, decahydronaphthyl, etc.), optionally substituted with one or more (such as 1 to 3) suitable substituents. The cycloalkyl group has 3 to 15, for example 3 to 10, 3 to 8, or 3 to 6 carbon atoms. For example, as used herein, the term “C3-C8 cycloalkyl” refers to a saturated or unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl) having 3 to 8 cyclic carbon atoms, which is optionally substituted with one or more (such as 1 to 3) suitable substituents, such as methyl-substituted cyclopropyl.

[0043] As used herein, the term "alkoxy" means an "alkyl" as defined above, which is attached to a parent molecule via an oxygen atom, such as a C1-C6 alkoxy or a C1-C3 alkoxy. Representative examples of C1-C6 alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy, hexoxy, etc., wherein the alkoxy group may optionally be substituted by one or more (such as 1 to 3) identical or different substituents.

[0044] As used herein, the terms “halogenated” or “halogen” refer to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0045] As used herein, the term "haloalkyl" refers to an alkyl group substituted with one or more (such as 1 to 3) identical or different halogen atoms. For example, the term "C1-C6 haloalkyl" as used herein refers to a haloalkyl group having 1 to 6 carbon atoms, including but not limited to -CH2F, -CHF2, -CF3, -CH2CF3, -CF2CF3, -CH2CH2CF3, -CH2Cl, etc. The haloalkyl group as used herein may optionally be substituted with one or more (such as 1 to 3) substituents described herein.

[0046] As used herein, the term “haloalkoxy” refers to a “haloalkyl” as defined above, such as a C1-C6 haloalkoxy, which is attached to a portion of the parent molecule by an oxygen atom.

[0047] As used herein, the term "heterocyclic group" refers to a monocyclic or polycyclic group having, for example, 2, 3, 4, 5, 6, 7, 8, or 9 carbon atoms in the ring and one or more (e.g., 1, 2, 3, or 4) groups selected from O, S, S(=O), S(=O)₂, N, and NR (R represents a hydrogen atom or a substituent, such as, but not limited to, alkyl or cycloalkyl). Heterocyclic groups include saturated and partially unsaturated non-aromatic heterocycles. Saturated heterocyclic groups may be called heterocyclic alkyl groups, such as 3-8-membered heterocyclic alkyl groups, 3-6-membered heterocyclic alkyl groups, 5-6-membered heterocyclic alkyl groups, etc. Unless otherwise specifically indicated in this specification, heterocyclic groups can be monocyclic, bicyclic, tricyclic, or more cyclic systems, which may include fused ring systems, fused ring systems, bridged ring systems, or spirocyclic systems. Specifically, 3-8 membered heterocyclic groups are groups having 3-8 carbon atoms and heteroatoms in the ring, for example, having 4 to 8, 4 to 7, 4 to 6, 5 to 8, 5 to 7, or 5 to 6 carbon atoms and heteroatoms (referred to as 4-8, 4-7, 4-6, 5-8, 5-7, and 5-6 membered heterocyclic groups, respectively), such as, but not limited to, ethylene oxide, aziridinyl, aziridine, aziridine-heptyl, oxaziridine-butyl, tetrahydrofuranyl, pyrrolylyl, pyrrolidone, imidazoyl, pyrazolylyl, tetrahydropyranyl, and aziridine-heptyl. The compounds include alkyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazine, trithianyl, etc.; as well as their cyclic derivatives, benzo[a] derivatives, heteroaryl[b] derivatives, or spirocyclic derivatives.

[0048] As used herein, the term "aryl" refers to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated π-electron system. Common aryl groups include (but are not limited to) phenyl, naphthyl, anthraceneyl, phenanthryl, acenaphthene, azulel, fluorenyl, indene, pyrene, etc. For example, the term "C6-C"... 10"Aryl" refers to an aromatic group containing 6 to 10 carbon atoms, such as phenyl or naphthyl. The aryl group may optionally be substituted by one or more (such as 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, -NO2, C1-C6 alkyl, etc.).

[0049] As used herein, the term "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic aromatic ring system containing at least one heteroatom selected from N, O, and S, having, for example, 5, 6, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly having 1, 2, 3, 4, 5, 6, 9, or 10 carbon atoms, and, in each case, may be benzofused. For example, as used herein, the term "5-10-membered heteroaryl" means a monocyclic, bicyclic, or tricyclic aromatic ring system having 5-10 ring atoms, and containing at least one heteroatom that may be the same or different (the heteroatom is, for example, N, O, or S). Examples of 5-10 membered heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrroleyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, and their benzo[a] derivatives; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and their benzo[a] derivatives. The heteroaryl group may optionally be substituted with one or more (e.g., one to three) suitable substituents (e.g., halogens, C1-C6 alkyl groups, etc.).

[0050] As used herein, the term "alkenyl" refers to a hydrocarbon group containing at least one C=C double bond. Alkenyl groups can be straight-chain or branched and contain 2 to 15 carbon atoms. For example, in this article, "C..." 2-6 "Alkenyl" refers to an alkenyl group containing 2 to 6 carbon atoms. Non-limiting examples of alkenyl groups include vinyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, octenyl, and decenyl. Alkenyl groups may be unsubstituted or substituted with one or more identical or different substituents.

[0051] As used herein, the term "alkynyl" refers to a hydrocarbon group having at least one C≡C triple bond. Alynyl groups can be straight-chain or branched and contain 2 to 15 carbon atoms. For example, in this article, "C..." 2-6 "Alynyl" refers to an alkynyl group containing 2 to 6 carbon atoms. Non-limiting examples of alkynyl groups include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, and 1,3-butadiynyl. The alkynyl group may be unsubstituted or substituted with one or more identical or different substituents.

[0052] The term "substitution" refers to the selective replacement of one or more (e.g., 1, 2, 3, 4, or 5) hydrogen atoms on a specified atom by a designated group, provided that the substitution does not exceed the normal valence of the specified atom in the present case and that the substitution forms a stable compound. Combinations of substituents and / or variables are permitted only if such combinations form a stable compound.

[0053] If a substituent is described as “optionally substituted with…”, then the substituent may be (1) unsubstituted or (2) substituted. If the carbon of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the carbon (to the extent that any hydrogens are present) may be substituted individually and / or together with independently selected substituents or not substituted. If the nitrogen of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the nitrogen (to the extent that any hydrogens are present) may each be substituted with independently selected substituents or not substituted.

[0054] If a substituent is described as being "independently selected" from a group of groups, then each substituent is selected independently of the others. Therefore, each substituent may be the same as or different from another (other) substituent.

[0055] As used herein, the term “one or more” means one or more under reasonable conditions, such as two, three, four, five, six, seven, eight, nine, or ten.

[0056] Unless otherwise specified, as used herein, the connection point of a substituent may be derived from any suitable location of the substituent.

[0057] When the bond of a substituent is such that it passes through the ring and connects two atoms, then such a substituent can be bonded to any cyclic atom in the substituted ring.

[0058] As used in this invention, the term "pharmaceutically acceptable" means that a substance or composition must be chemically and / or toxicologically compatible with other components constituting the formulation and / or the mammals treated therewith.

[0059] Pharmaceutically acceptable salts of the compounds of the present invention include their acid addition salts and base addition salts. Suitable acid addition salts are formed by acids that form pharmaceutically acceptable salts. Suitable base addition salts are formed by bases that form pharmaceutically acceptable salts. A review of suitable salts can be found in Stahl and Wermuth's "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

[0060] The term "stereoisomer" (or "optical isomer") refers to a stable isomer that has a perpendicular asymmetric plane due to having at least one chiral element (including a chiral center, chiral axis, chiral plane, etc.), thereby enabling the rotation of plane-polarized light. Since the compounds of this invention contain asymmetric centers and other chemical structures that may lead to stereoisomerism, this invention also includes these stereoisomers and mixtures thereof. Because the compounds of this invention (or pharmaceutically acceptable salts thereof) comprise asymmetric carbon atoms, they can exist as single stereoisomers, racemates, or mixtures of enantiomers and diastereomers. Typically, these compounds can be prepared as racemates. However, if desired, such compounds can be prepared or isolated to obtain pure stereoisomers, i.e., single enantiomers or diastereomers, or mixtures enriched with single stereoisomers (purity ≥98%, ≥95%, ≥93%, ≥90%, ≥88%, ≥85%, or ≥80%). As described below, a single stereoisomer of a compound is synthesized from an optically active starting material containing the desired chiral center, or prepared by separating or resolving a mixture of enantiomers, for example, by converting it into a mixture of diastereomers and then separating or recrystallizing, chromatographically processing, using chiral resolving reagents, or by directly separating the enantiomers on a chiral chromatographic column. Starting compounds with specific stereochemistry are commercially available or prepared according to the methods described below and then resolved by methods well known in the art. The term "enantiomer" refers to a pair of stereoisomers that are mirror images of each other. The term "diastereomer" or "diastereomer" refers to an optically active isomer that is not mirror images of each other. The term "racemic mixture" or "racemate" refers to a mixture containing equal parts of a single enantiomer (i.e., an equimolar mixture of two R and S enantiomers). The term "non-racemic mixture" refers to a mixture containing unequal parts of a single enantiomer. Unless otherwise stated, all stereoisomers of the compounds of this invention are within the scope of this invention.

[0061] The term "tautomer" (or "tautomer form") refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium can be achieved in the tautomer. For example, proton tautomers (or proton transfer tautomers) include (but are not limited to) interconversions via proton transfer, such as keto-enol isomerization, imine-enamine isomerization, amide-imine alcohol isomerization, etc. Unless otherwise stated, all tautomer forms of the compounds of this invention are within the scope of this invention.

[0062] The term "polymorph" (or "polymorphic form") refers to the solid crystalline form of a compound or complex. Polymorphs can be detected, classified, and identified using well-known techniques, including (but not limited to) differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray powder diffraction (XRPD), single-crystal X-ray diffraction (SCXRD), solid-state nuclear magnetic resonance (NMR), infrared spectroscopy (IR), Raman spectroscopy, and scanning electron microscopy (SEM).

[0063] The term "solvent" refers to a substance formed by the combination of the compound of the present invention (or a pharmaceutically acceptable salt thereof) with at least one solvent molecule through non-covalent intermolecular forces.

[0064] The term "nitrogen oxides" refers to compounds formed by the oxidation of nitrogen atoms in the structure of tertiary amines or nitrogen-containing (aromatic) heterocyclic compounds.

[0065] This invention also includes all pharmaceutically acceptable isotopically labeled compounds that are identical to the compounds of this invention, except that one or more atoms are replaced by atoms having the same atomic number but with an atomic mass or mass number different from the dominant atomic mass or mass number in nature. Examples of isotopes suitable for inclusion in the compounds of this invention include, but are not limited to, isotopes of hydrogen (e.g., 2 H, 3 H, deuterium (D), tritium (T); carbon isotopes (e.g., H, deuterium (D), tritium (T)); 11 C 13 C and 14 C); isotopes of chlorine (e.g.) 37 Cl); isotopes of fluorine (e.g., Cl); 18 F); isotopes of iodine (e.g., F); 123 I and 125 I); nitrogen isotopes (e.g.) 13 N and 15 N); isotopes of oxygen (e.g., N); 15 O、 17 O and 18 O); and isotopes of sulfur (e.g. 35 S).

[0066] The scope of this invention also includes metabolites of the compounds of this invention, i.e., substances formed in the body when the compounds of this invention are administered. Such products can be generated, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compound. Therefore, this invention includes metabolites of the compounds of this invention, including compounds obtained by methods that expose the compounds of this invention to mammals for a time sufficient to produce their metabolites.

[0067] This invention further includes, within its scope, prodrugs of the compounds of the invention, which are certain derivatives of the compounds of the invention that may themselves have little or no pharmacological activity, and which, when administered to or onto the body, can be converted, for example, by hydrolysis and cleavage into the compounds of the invention having the desired activity. Typically, such prodrugs are functional group derivatives of the compounds that are readily converted in vivo into the compounds with the desired therapeutic activity. Further information regarding the use of prodrugs can be found in “Pro-drugs as Novel Delivery Systems,” Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella) and “Bioreversible Carriers in Drug Design,” Pergamon Press, 1987 (EB Roche, editor, American Pharmaceutical Association). The prodrugs of the invention can be prepared, for example, by replacing suitable functional groups present in the compounds of the invention with certain portions known to those skilled in the art as “pro-moiety” (e.g., as described in “Design of Prodrugs,” H. Bundgaard (Elsevier, 1985)).

[0068] The term "independently" means that at least two groups (or ring systems) in a structure with the same or similar value ranges can have the same or different meanings under specific circumstances. For example, if substituent X and substituent Y are independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when substituent X is hydrogen, substituent Y can be hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl; similarly, when substituent Y is hydrogen, substituent X can be hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl.

[0069] This invention also covers compounds of the invention containing protecting groups. In any process of preparing the compounds of the invention, protection of sensitive or reactive groups on any relevant molecule may be necessary and / or desired, thereby forming a form of chemical protection for the compounds of the invention. This can be achieved by conventional protecting groups, for example, those described in *Protective Groups in Organic Chemistry*, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene & P. ​​G.W. Uts, *Protective Groups in Organic Synthesis*, John Wiley & Sons, 1991, which are incorporated herein by reference. Protecting groups can be removed at appropriate subsequent stages using methods known in the art.

[0070] When the term "about" is used in this invention to modify a numerical value or range, it refers to the numerical value or range and an acceptable error range for those skilled in the art, such as ±10%, ±5%, ±4%, ±3%, ±2%, ±1%, ±0.5%, etc.

[0071] compound

[0072] One object of the present invention is to provide a compound of formula I or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof.

[0073]

[0074] in,

[0075] R 1 Selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic groups, C6-C 10 aryl and 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic, C6-C 10 aryl or 5-10 heteroaryl groups are optionally surrounded by one or more R groups. 5 replace;

[0076] R 5 Each occurrence is independently selected from halogens, -CN, -OH, -NH2, -NO2, and -S(O). p R A -COR A=O, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 alkoxy, 3-8 membered heterocyclic, C2-C6 alkenyl, C2-C6 alkynyl, C6-C 10 aryl and 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 alkoxy, 3-8 membered heterocyclic, C2-C6 alkenyl, C2-C6 alkynyl, C6-C 10 aryl or 5-10 heteroaryl groups are optionally surrounded by one or more R groups. 6 replace;

[0077] R 6 Each occurrence is independently selected from halogens, -CN, -OH, -NH2, -NO2, and -S(O). p R A -COR A S = O;

[0078] R A Independently selected from H and C1-C6 alkyl groups;

[0079] p is selected from 1 and 2;

[0080] R 2 Each of the elements, when appearing independently, is selected from H, -OH, halogen, -CN, -NH2, -NO2, C1-C6 alkyl, C1-C6 alkoxy, and C3-C8 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, or C3-C8 cycloalkyl is optionally separated by one or more R. 7 replace;

[0081] R 7 Each of these elements is independently selected from -OH, halogen, -CN, -NH2, -NO2, C1-C6 alkoxy, C1-C6 alkyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy in each occurrence.

[0082] m is selected from 0, 1, 2, 3, and 4;

[0083] R 3 Selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic groups, C6-C 10 aryl and 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, 3-8 membered heterocyclic, C6-C 10 aryl or 5-10 heteroaryl groups are optionally surrounded by one or more R groups. 8 replace;

[0084] R 8Each of these elements is independently selected from -OH, halogen, -CN, -NH2, -NO2, C1-C6 alkoxy, C1-C6 alkyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy in each occurrence.

[0085] R 4 Each of the following groups, when appearing independently, is selected from H, -OH, halogen, CN, -NH2, -NO2, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, and 3-8 membered heterocyclic groups, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, or 3-8 membered heterocyclic group is optionally replaced by one or more R groups. 9 replace;

[0086] R 9 Each of these groups is independently selected from -OH, halogen, -CN, -NH2, -NO2, C1-C6 alkoxy, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy, and 3-8 membered heterocyclic groups when it appears.

[0087] n is selected from 0, 1, and 2; and

[0088] X is selected from CH and N.

[0089] According to some embodiments of the present invention, R 1 Selected from C1-C6 alkyl, C3-C8 cycloalkyl, and 3-8 membered heterocyclic groups, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, or 3-8 membered heterocyclic group is optionally surrounded by one or more R groups. 5 replace.

[0090] According to some embodiments of the present invention, R 5 Each time it appears, it is independently selected from halogens, -CN, -OH, and -S(O). p R A C3-C8 cycloalkyl and C6-C 10 Aryl, the C3-C8 cycloalkyl or C6-C 10 Aryl group is optionally bounded by one or more R 6 replace.

[0091] According to some embodiments of the present invention, R 6 Each time it appears, it is independently selected from halogens and =O.

[0092] According to some embodiments of the present invention, R A It is a C1-C6 alkyl group.

[0093] According to some embodiments of the present invention, p is 2.

[0094] According to some embodiments of the present invention, R 1Selected from C1-C6 alkyl, C3-C8 cycloalkyl, and 3-8 membered heterocyclic groups, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, or 3-8 membered heterocyclic group is optionally surrounded by one or more R groups. 5 replace;

[0095] R 5 Each time it appears, it is independently selected from halogens, -CN, -OH, and -S(O). p R A C3-C8 cycloalkyl and C6-C 10 Aryl, the C3-C8 cycloalkyl or C6-C 10 Aryl group is optionally bounded by one or more R 6 replace;

[0096] R 6 Each time it appears, it is independently selected from halogens and =O;

[0097] R A It is a C1-C6 alkyl group; and

[0098] p is 2.

[0099] In some embodiments of the present invention, R 1 The group is selected from C1-C6 alkyl, C3-C6 cycloalkyl and 3-6 heterocyclic alkyl, wherein the C1-C6 alkyl, C3-C6 cycloalkyl or 3-6 heterocyclic alkyl is optionally substituted by one or more groups each independently selected from: halogen, -CN, -OH, -S(O)2 (C1-C6 alkyl), C3-C6 cycloalkyl optionally substituted with halogen or =O, and phenyl optionally substituted with halogen.

[0100] In some embodiments of the present invention, R 1 The group is selected from C1-C6 alkyl, C3-C6 cycloalkyl and 3-6 oxocyclic alkyl, wherein the C1-C6 alkyl, C3-C6 cycloalkyl or 3-6 oxocyclic alkyl is optionally substituted by one or more groups each independently selected from: halogen, -CN, -OH, -S(O)2 (C1-C6 alkyl), C3-C6 cycloalkyl optionally substituted with halogen or =O, and phenyl optionally substituted with halogen.

[0101] In some embodiments of the present invention, R 1 The group is selected from C1-C6 alkyl, C3-C6 cycloalkyl, oxetyl, tetrahydrofuranyl, and tetrahydropyranyl, wherein the C1-C6 alkyl or C3-C6 cycloalkyl is optionally substituted with a group selected from: halogen, -CN, -OH, -S(O)2 (C1-C6 alkyl), C3-C6 cycloalkyl optionally substituted with halogen or =O, and phenyl optionally substituted with halogen.

[0102] In some embodiments of the present invention, R 1 The group is selected from C1-C6 alkyl, C3-C6 cycloalkyl, oxetyl, tetrahydrofuranyl and tetrahydropyranyl, wherein the C1-C6 alkyl or C3-C6 cycloalkyl is optionally substituted with a group selected from the following: -F, -CN, -OH, -S(O)2CH3, C3-C6 cycloalkyl, oxocyclohexyl and fluorophenyl.

[0103] In some embodiments of the present invention, R 1 Selected from

[0104] In some embodiments of the present invention, R 1 Selected from

[0105] Among them, wavy lines This indicates the connection point between the group and the rest of the molecule.

[0106] According to some embodiments of the present invention, R 2 Each time it appears, it is independently selected from H and halogen.

[0107] In some embodiments of the present invention, R 2 Each time it appears, it is independently selected from H and F.

[0108] According to some embodiments of the present invention, m is 1.

[0109] According to some embodiments of the present invention, R 3 Selected from C6-C 10 Aryl and 5-10 heteroaryl, the C6-C 10 aryl or 5-10 heteroaryl groups are optionally surrounded by one or more R groups. 8 replace.

[0110] According to some embodiments of the present invention, R 8 Each of the elements, when appearing independently, is selected from halogens, C1-C6 alkyl groups, and C1-C6 haloalkyl groups; preferably, R 8 Each of these elements is independently selected from F, Cl, -CF3, and -CH3 each time it appears.

[0111] According to some embodiments of the present invention, R 3 Selected from C6-C 10 Aryl and 5-10 heteroaryl, the C6-C 10 The aryl or 5-10 heteroaryl group is optionally substituted by one or more groups, each independently selected from the following groups: halogen, C1-C6 alkyl and C1-C6 haloalkyl.

[0112] In some embodiments of the present invention, R 3 The group is selected from phenyl and 5-6 heteroaryl groups, wherein the phenyl or 5-6 heteroaryl group is optionally substituted by one or more groups, each independently selected from the group consisting of halogen, C1-C6 alkyl and C1-C6 haloalkyl.

[0113] In some embodiments of the present invention, R 3 The group is selected from phenyl and 5-6 nitrogen-containing heteroaryl groups, wherein the phenyl or 5-6 nitrogen-containing heteroaryl group is optionally substituted by one or more groups, each independently selected from the group consisting of halogen, C1-C6 alkyl and C1-C6 haloalkyl.

[0114] In some embodiments of the present invention, R 3 Selected from phenyl and pyridyl, wherein the phenyl or pyridyl group is optionally substituted by one or more groups each independently selected from the following groups: halogen, C1-C6 alkyl and C1-C6 haloalkyl.

[0115] In some embodiments of the present invention, R 3 Selected from phenyl and pyridyl, wherein the phenyl or pyridyl group is optionally substituted by one or more groups, each independently selected from the following: F, Cl, -CF3 and -CH3.

[0116] In some embodiments of the present invention, R 3 Selected from:

[0117] Among them, wavy lines This indicates the connection point between the group and the rest of the molecule.

[0118] According to some embodiments of the present invention, R 4 Each group is independently selected from C1-C6 alkyl and C3-C8 cycloalkyl groups in each occurrence, wherein the C1-C6 alkyl group is optionally substituted by one or more groups independently selected from C1-C6 alkoxy and 3-8 membered heterocyclic groups.

[0119] In some embodiments of the present invention, R 4 Each of the components is independently selected from C1-C6 alkyl and C3-C8 cycloalkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with C1-C6 alkoxy groups or 5-6 member oxygen-containing heterocyclic alkyl groups.

[0120] In some embodiments of the present invention, R 4 Each of the components is independently selected from C1-C6 alkyl and C3-C8 cycloalkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with C1-C6 alkoxy or tetrahydropyranyl groups.

[0121] In some embodiments of the present invention, R 4Each time it appears, it is independently selected from isopropyl...

[0122] Among them, wavy lines This indicates the connection point between the group and the rest of the molecule.

[0123] According to some embodiments of the present invention, n is 1.

[0124] This invention covers compounds of formula I obtained by any combination of the above-mentioned preferred groups.

[0125] According to some embodiments of the present invention, the compound is a compound of formula I-1:

[0126]

[0127] in,

[0128] R 1 R 2 R 3 R 4 m and n are as defined above.

[0129] This invention covers compounds of formula I-1 obtained by any combination of the above-mentioned preferred groups.

[0130] According to some embodiments of the present invention, the compound is a compound of formula I-2:

[0131]

[0132]

[0133] in,

[0134] R 1 R 2 R 3 R 4 m and n are as defined above.

[0135] This invention covers compounds of formula I-2 obtained by any combination of the above-mentioned preferred groups.

[0136] According to some embodiments of the present invention, the compound is a compound of formula I-3:

[0137]

[0138] in,

[0139] R 1 R 2 R 4 R 8X, m, and n are as defined above;

[0140] q is selected from 0, 1, 2, 3, 4 and 5.

[0141] In some embodiments of the present invention, q is 1.

[0142] This invention covers compounds of formula I-3 obtained by any combination of the above-mentioned preferred groups.

[0143] According to some embodiments of the present invention, the compound is a compound of formula I-4:

[0144]

[0145] in,

[0146] R 1 R 2 R 4 R 8 X, m, and n are as defined above;

[0147] r is selected from 0, 1, 2, 3, and 4.

[0148] In some embodiments of the present invention, r is 1.

[0149] This invention covers compounds of formulas I-4 obtained by any combination of the above-mentioned preferred groups.

[0150] According to some embodiments of the present invention, the compounds of the present invention are selected from:

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157] Preparation method

[0158] Another object of the present invention is to provide a method for preparing the compounds of the present invention, comprising the steps shown in reaction route 1 below:

[0159] Reaction route 1

[0160]

[0161] in,

[0162] R 1 R 2 R 3 R 4 X, m, and n are as defined above.

[0163] The reaction is preferably carried out in the presence of a suitable organic or inorganic base. The organic or inorganic base may be selected from diisopropylethylamine, triethylamine, sodium carbonate, potassium carbonate, and cesium carbonate, with diisopropylethylamine being preferred.

[0164] The reaction is preferably carried out in the presence of a suitable condensing agent, which may be selected from DCC, EDCI, HOBt, BOP, PyBOP, HATU, HBTU, with HATU being preferred.

[0165] The reaction is carried out in a suitable organic solvent. The organic solvent may be selected from dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, and any combination thereof, with N,N-dimethylformamide being preferred.

[0166] The reaction is carried out at a suitable temperature. The preferred temperature is 20-80°C.

[0167] The reaction should proceed for an appropriate time, such as 2-24 hours.

[0168] In some implementations, the intermediate IB is the following intermediate I-B1:

[0169]

[0170] On the other hand, the present invention provides a method for synthesizing intermediates IA and IB, wherein intermediate IA can be synthesized by the method shown in reaction route 2 below:

[0171] Reaction route 2

[0172]

[0173] in,

[0174] LG represents a leaving group, which includes, but is not limited to, halogen atoms, methanesulfonyloxy groups, trifluoromethanesulfonates, etc.; and

[0175] R 1 R 2 X and m are as defined above for compounds of formula I.

[0176] Step 1: React compound IAI with R 1 -LG reaction to give compound IA-II

[0177] The reaction is preferably carried out in the presence of a suitable organic or inorganic base. The organic or inorganic base may be selected from diisopropylethylamine, triethylamine, sodium carbonate, potassium carbonate, and cesium carbonate, with cesium carbonate being preferred.

[0178] The reaction is carried out in a suitable organic solvent. The organic solvent may be selected from dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, and any combination thereof, with N,N-dimethylformamide being preferred.

[0179] The reaction is carried out at a suitable temperature. The preferred temperature is 80-90°C.

[0180] The reaction should proceed for an appropriate time, such as 2-24 hours.

[0181] Step 2: React compound IA-II with p-aminophenylboronic acid ester to obtain compound IA.

[0182] The reaction is preferably carried out in the presence of a metal catalyst and a base. The metal catalyst is preferably a palladium catalyst, such as tris(dibenzylacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, triphenylphosphine palladium, palladium acetate, and preferably [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride. The base is preferably an inorganic base, such as potassium phosphate, potassium carbonate, cesium carbonate, sodium carbonate, sodium bicarbonate, or potassium bicarbonate, and preferably sodium carbonate.

[0183] The reaction is carried out in a suitable solvent. The solvent may be selected from N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1,4-dioxane, and any combination thereof, preferably a mixture of 1,4-dioxane and water.

[0184] The reaction is carried out at a suitable temperature. The preferred temperature is 60-100°C.

[0185] The reaction should proceed for an appropriate time, such as 2-8 hours.

[0186] Alternatively, intermediate IA can also be synthesized via the method shown in reaction route 3 below:

[0187] Reaction route 3

[0188]

[0189] in,

[0190] R 1 R 2 X and m are as defined above for compounds of formula I.

[0191] Step 1: React compound IAI with R 1The OH reaction yields compound IA-II.

[0192] The reaction is preferably carried out in the presence of a phosphine catalyst and an ester. The phosphine catalyst is preferably a phosphine ligand catalyst, such as triphenylphosphine, tributylphosphine, trihexylphosphine, or tris(2-methylphenyl)phosphine, with triphenylphosphine being the most preferred. The ester is preferably an azodicarbonate, such as diethyl azodicarbonate, diisopropyl azodicarbonate, di-tert-butyl azodicarbonate, or bis(4-chlorobenzyl)azodicarbonate, with diethyl azodicarbonate being the most preferred.

[0193] The reaction is carried out in a suitable solvent. The solvent may be selected from tetrahydrofuran, diethyl ether, toluene, N,N-dimethylformamide, and any combination thereof, with tetrahydrofuran being preferred.

[0194] The reaction is carried out at a suitable temperature. The preferred temperature is 25-70°C.

[0195] The reaction should proceed for an appropriate time, such as 8-16 hours.

[0196] Step 2: React compound IA-II with p-aminophenylboronic acid ester to obtain compound IA.

[0197] The reaction is preferably carried out in the presence of a metal catalyst and a base. The metal catalyst is preferably a palladium catalyst, such as tris(dibenzylacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, triphenylphosphine palladium, palladium acetate, and preferably [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride. The base is preferably an inorganic base, such as potassium phosphate, potassium carbonate, cesium carbonate, sodium carbonate, sodium bicarbonate, or potassium bicarbonate, and preferably sodium carbonate.

[0198] The reaction is carried out in a suitable solvent. The solvent may be selected from N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1,4-dioxane, and any combination thereof, preferably a mixture of 1,4-dioxane and water.

[0199] The reaction is carried out at a suitable temperature. The preferred temperature is 60-100°C.

[0200] The reaction should proceed for an appropriate time, such as 2-8 hours.

[0201] Alternatively, intermediate IA can also be synthesized via the method shown in reaction route 4 below:

[0202] Reaction route 4

[0203]

[0204] in,

[0205] LG represents a leaving group, which includes, but is not limited to, halogen atoms, methanesulfonyloxy groups, trifluoromethanesulfonates, etc.; and

[0206] R 1 R 2 X and m are as defined above for compounds of formula I.

[0207] Step 1: React compound IA-III with R 1 -LG reaction to give compound IA-IV

[0208] The reaction is preferably carried out in the presence of a suitable organic or inorganic base. The organic or inorganic base may be selected from diisopropylethylamine, triethylamine, sodium carbonate, potassium carbonate, and cesium carbonate, with cesium carbonate being preferred.

[0209] The reaction is carried out in a suitable organic solvent. The organic solvent may be selected from dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, and any combination thereof, with N,N-dimethylformamide being preferred.

[0210] The reaction is carried out at a suitable temperature. The preferred temperature is 80-90°C.

[0211] The reaction should proceed for an appropriate time, such as 2-24 hours.

[0212] Step 2: Compound IA-IV is subjected to a substitution reaction to obtain compound IA-II.

[0213] The reaction is preferably carried out in the presence of a suitable base. The preferred base is ammonia.

[0214] The reaction is carried out in a suitable organic solvent. The organic solvent may be selected from dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, 1,4-dioxane, and any combination thereof, with 1,4-dioxane being preferred.

[0215] The reaction is carried out at a suitable temperature. The preferred temperature is 80-90°C.

[0216] The reaction is carried out under suitable conditions. Preferably, these conditions are microwave heating.

[0217] The reaction should proceed for an appropriate time, such as 2-24 hours.

[0218] Step 3: React compound IA-II with p-aminophenylboronic acid ester to obtain compound IA.

[0219] The reaction is preferably carried out in the presence of a metal catalyst and a base. The metal catalyst is preferably a palladium catalyst, such as tris(dibenzylacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, triphenylphosphine palladium, palladium acetate, and preferably [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride. The base is preferably an inorganic base, such as potassium phosphate, potassium carbonate, cesium carbonate, sodium carbonate, sodium bicarbonate, or potassium bicarbonate, and preferably sodium carbonate.

[0220] The reaction is carried out in a suitable solvent. The solvent may be selected from N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1,4-dioxane, and any combination thereof, preferably a mixture of 1,4-dioxane and water.

[0221] The reaction is carried out at a suitable temperature. The preferred temperature is 60-100°C.

[0222] The reaction should proceed for an appropriate time, such as 2-8 hours.

[0223] Alternatively, intermediate IA can also be synthesized via the method shown in reaction route 5 below:

[0224] Reaction route 5

[0225]

[0226] in,

[0227] R 1 R 2 X and m are as defined above for compounds of formula I.

[0228] Step 1: React compound IA-III with R 1 The OH reaction yields compounds IA-IV.

[0229] The reaction is preferably carried out in the presence of a phosphine catalyst and an ester. The phosphine catalyst is preferably a phosphine ligand catalyst, such as triphenylphosphine, tributylphosphine, trihexylphosphine, or tris(2-methylphenyl)phosphine, with triphenylphosphine being the most preferred. The ester is preferably an azodicarbonate, such as diethyl azodicarbonate, diisopropyl azodicarbonate, di-tert-butyl azodicarbonate, or bis(4-chlorobenzyl)azodicarbonate, with diethyl azodicarbonate being the most preferred.

[0230] The reaction is carried out in a suitable solvent. The solvent may be selected from tetrahydrofuran, diethyl ether, toluene, N,N-dimethylformamide, and any combination thereof, with tetrahydrofuran being preferred.

[0231] The reaction is carried out at a suitable temperature. The preferred temperature is 25-70°C.

[0232] The reaction should proceed for an appropriate time, such as 8-16 hours.

[0233] Step 2: Compound IA-IV is subjected to a substitution reaction to obtain compound IA-II.

[0234] The reaction is preferably carried out in the presence of a suitable base. The preferred base is ammonia.

[0235] The reaction is carried out in a suitable organic solvent. The organic solvent may be selected from dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, 1,4-dioxane, and any combination thereof, with 1,4-dioxane being preferred.

[0236] The reaction is carried out at a suitable temperature. The preferred temperature is 80-90°C.

[0237] The reaction is carried out under suitable conditions. Preferably, these conditions are microwave heating.

[0238] The reaction should proceed for an appropriate time, such as 2-24 hours.

[0239] Step 3: React compound IA-II with p-aminophenylboronic acid ester to obtain compound IA.

[0240] The reaction is preferably carried out in the presence of a metal catalyst and a base. The metal catalyst is preferably a palladium catalyst, such as tris(dibenzylacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, triphenylphosphine palladium, palladium acetate, and preferably [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride. The base is preferably an inorganic base, such as potassium phosphate, potassium carbonate, cesium carbonate, sodium carbonate, sodium bicarbonate, or potassium bicarbonate, and preferably sodium carbonate.

[0241] The reaction is carried out in a suitable solvent. The solvent may be selected from N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1,4-dioxane, and any combination thereof, preferably a mixture of 1,4-dioxane and water.

[0242] The reaction is carried out at a suitable temperature. The preferred temperature is 60-100°C.

[0243] The reaction should proceed for an appropriate time, such as 2-8 hours.

[0244] Intermediate I-B1 can be synthesized via the method shown in reaction route 6 below:

[0245] Reaction route 6

[0246]

[0247] Step 1: React compound IBI with R3 -NHNH2 reaction to give compound IB-II

[0248] The reaction is carried out in the presence of a suitable solvent. The solvent may be selected from methanol, ethanol, tetrahydrofuran, N,N-dimethylformamide, and any combination thereof, with methanol being preferred.

[0249] The reaction is carried out at a suitable temperature. The preferred temperature is 25-70°C.

[0250] The reaction should proceed for an appropriate time, such as 2-8 hours.

[0251] Step 2: Deprotect compound IB-II to obtain compound IB-III.

[0252] The reaction is preferably carried out in the presence of a catalyst. The catalyst is preferably an acid catalyst, such as hydrochloric acid, trifluoroacetic acid, acetic acid, or camphor sulfonic acid.

[0253] The reaction is carried out at a suitable temperature. The preferred temperature is 25-70°C.

[0254] The reaction is carried out in a suitable solvent. The solvent may be selected from water, 1,4-dioxane, tetrahydrofuran, N,N-dimethylformamide, and any combination thereof, with methanol being preferred.

[0255] The reaction should proceed for an appropriate time, such as 4-16 hours.

[0256] Step 3: React compound IB-III with cyclo(isopropyl)malonate to obtain compound IB-IV.

[0257] The reaction is preferably carried out in the presence of an acid. The acid is preferably a Lewis acid, such as tetrahydropyrrole acetate or pyridine p-toluenesulfonate, with tetrahydropyrrole acetate being more preferred.

[0258] The reaction is carried out in a suitable solvent. The solvent may be selected from tetrahydrofuran, diethyl ether, toluene, N,N-dimethylformamide, and any combination thereof, with toluene being preferred.

[0259] The reaction is carried out at a suitable temperature. The preferred temperature is 25-70°C.

[0260] The reaction should proceed for an appropriate time, such as 8-16 hours.

[0261] Step 4: Cyclic closure and hydrolysis of compound IB-IV to obtain compound I-B1

[0262] The reaction is preferably carried out in the presence of a base. The base is preferably an organic base, such as sodium methoxide, sodium ethoxide, sodium tert-butoxide, or potassium tert-butoxide, with sodium methoxide being the most preferred.

[0263] The reaction is carried out in a suitable solvent. The solvent may be selected from methanol, ethanol, tert-butanol, 1,4-dioxane, and any combination thereof, with methanol being preferred.

[0264] The reaction is carried out at a suitable temperature. The preferred temperature is 25-70°C.

[0265] The reaction should proceed for an appropriate time, such as 4-16 hours.

[0266] Intermediate I-B1 can also be synthesized via the method shown in reaction route 7 below:

[0267] Reaction route 7

[0268]

[0269] Step 1: React compound IBV with dimethyl malonate to obtain compound IB-VI;

[0270] The reaction is preferably carried out in the presence of a base. The base is preferably an organic or inorganic base, such as sodium hydride, sodium methoxide, sodium ethoxide, or potassium carbonate, with sodium hydride being the most preferred.

[0271] The reaction is carried out in a suitable solvent. The solvent may be selected from tetrahydrofuran, diethyl ether, N,N-dimethylformamide, and any combination thereof, with tetrahydrofuran being preferred.

[0272] The reaction is carried out at a suitable temperature. The preferred temperature is 0-25°C.

[0273] The reaction should proceed for an appropriate time, such as 2-8 hours.

[0274] Step 2: React compound IB-VI with R 3 -NHNH2 reaction to give compound IB-VII;

[0275] The reaction is preferably carried out in the presence of a base. The base is preferably an organic or inorganic base, such as sodium acetate, sodium carbonate, or potassium carbonate, with sodium acetate being the most preferred.

[0276] The reaction is carried out at a suitable temperature. The preferred temperature is 50-90°C.

[0277] The reaction is carried out in a suitable solvent. The solvent may be selected from acetic acid, toluene, N,N-dimethylformamide, dimethyl sulfoxide, and any combination thereof, with acetic acid being preferred.

[0278] The reaction should proceed for an appropriate time, such as 4-16 hours.

[0279] Step 3: Oxidize compound IB-VII to obtain compound IB-VIII;

[0280] The reaction is carried out in the presence of an oxidizing agent. The oxidizing agent is preferably an inorganic salt, such as copper chloride or palladium on carbon, with copper chloride being more preferred.

[0281] The reaction is carried out in a suitable solvent. The solvent may be selected from tetrahydrofuran, diethyl ether, toluene, N,N-dimethylformamide, and any combination thereof, with toluene being preferred.

[0282] The reaction is carried out at a suitable temperature, preferably 25-70°C.

[0283] The reaction should proceed for an appropriate time, such as 8-16 hours.

[0284] Step 4: Hydrolyze compound IB-VIII to obtain compound I-B1;

[0285] The reaction is preferably carried out in the presence of a base. The base is preferably an inorganic base, such as lithium hydroxide, sodium hydroxide, potassium hydroxide, or potassium carbonate, with lithium hydroxide being the most preferred.

[0286] The reaction is carried out in a suitable solvent. The solvent may be selected from tetrahydrofuran, methanol, ethanol, water, or any combination thereof, preferably a mixture of methanol and water.

[0287] The reaction is carried out at a suitable temperature. The preferred temperature is 25-60°C.

[0288] The reaction should proceed for an appropriate time, such as 2-8 hours.

[0289] Some compounds of general formula I can be synthesized by the method shown in reaction route 8 below:

[0290] Reaction route 8

[0291]

[0292] in,

[0293] LG represents a leaving group, which includes, but is not limited to, halogen atoms, methanesulfonyloxy groups, trifluoromethanesulfonates, etc.; and

[0294] R 1 R 2 R 3 R 4 X, m, and n are as defined above for compounds of formula I.

[0295] Step 1: React compound IAI with p-methoxybenzyl chloride to obtain compound IAV.

[0296] The reaction is preferably carried out in the presence of a suitable organic or inorganic base. The organic or inorganic base may be selected from diisopropylethylamine, triethylamine, sodium carbonate, potassium carbonate, and cesium carbonate, with potassium carbonate being preferred.

[0297] The reaction is carried out in a suitable organic solvent. The organic solvent may be selected from dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, and any combination thereof, with N,N-dimethylformamide being preferred.

[0298] The reaction is carried out at a suitable temperature. The preferred temperature is 20-30°C.

[0299] The reaction should proceed for an appropriate time, such as 2-24 hours.

[0300] Step 2: React compound IAV with p-aminophenylboronic acid ester to obtain compound IA-VI.

[0301] The reaction is preferably carried out in the presence of a metal catalyst and a base. The metal catalyst is preferably a palladium catalyst, such as tris(dibenzylacetone)dipalladium, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, triphenylphosphine palladium, palladium acetate, and preferably [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride. The base is preferably an inorganic base, such as potassium phosphate, potassium carbonate, cesium carbonate, sodium carbonate, sodium bicarbonate, or potassium bicarbonate, and preferably sodium carbonate.

[0302] The reaction is carried out in a suitable solvent. The solvent may be selected from N,N-dimethylformamide, N-methylpyrrolidone, toluene, ethanol, ethylene glycol dimethyl ether, water, 1,4-dioxane, and any combination thereof, preferably a mixture of 1,4-dioxane and water.

[0303] The reaction is carried out at a suitable temperature. The preferred temperature is 60-100°C.

[0304] The reaction should proceed for an appropriate time, such as 2-8 hours.

[0305] Step 3: React compound IA-VI with IB to obtain compound IA-VII.

[0306] The reaction is preferably carried out in the presence of a suitable organic or inorganic base. The organic or inorganic base may be selected from diisopropylethylamine, triethylamine, sodium carbonate, potassium carbonate, and cesium carbonate, with diisopropylethylamine being preferred.

[0307] The reaction is preferably carried out in the presence of a suitable condensing agent, which may be selected from DCC, EDCI, HOBt, BOP, PyBOP, HATU, HBTU, with HATU being preferred.

[0308] The reaction is carried out in a suitable organic solvent. The organic solvent may be selected from dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, and any combination thereof, with N,N-dimethylformamide being preferred.

[0309] The reaction is carried out at a suitable temperature. The preferred temperature is 20-80°C.

[0310] The reaction should proceed for an appropriate time, such as 2-24 hours.

[0311] Step 4: Compound IA-VII is subjected to a deprotection reaction to obtain compound IA-VIII.

[0312] The reaction is carried out in a suitable acid. The acid may be selected from trifluoroacetic acid, methanesulfonic acid, hydrochloric acid, or any combination thereof, with trifluoroacetic acid being preferred.

[0313] The reaction is carried out at a suitable temperature. The preferred temperature is 20-80°C.

[0314] The reaction should proceed for an appropriate time, such as 2-24 hours.

[0315] Step 5: React compound IA-VIII with R 1 -LG reaction to give compound I

[0316] The reaction is preferably carried out in the presence of a suitable organic or inorganic base. The organic or inorganic base may be selected from diisopropylethylamine, triethylamine, sodium carbonate, potassium carbonate, and cesium carbonate, with cesium carbonate being preferred.

[0317] The reaction is carried out in a suitable organic solvent. The organic solvent may be selected from dichloromethane, N,N-dimethylformamide, dimethyl sulfoxide, and any combination thereof, with N,N-dimethylformamide being preferred.

[0318] The reaction is carried out at a suitable temperature. The preferred temperature is 80-90°C.

[0319] The reaction should proceed for an appropriate time, such as 2-24 hours.

[0320] As used herein, the term "suitable" means that the choice of a particular compound or conditions will depend on the specific synthetic operation to be performed and the characteristics of one or more molecules to be transformed, but such choice is within the capabilities of those skilled in the art. All process / method steps described herein are performed under conditions sufficient to provide the products shown. Those skilled in the art will understand that all reaction conditions (including, for example, reaction solvents, reaction times, reaction temperatures, and whether the reaction should be carried out under an anhydrous or inert atmosphere, etc.) can be varied to optimize the desired product yield, and such variations are within the capabilities of those skilled in the art.

[0321] Pharmaceutical Compositions and Kits

[0322] Another object of the present invention is to provide a pharmaceutical composition comprising a preventive or therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug, and one or more pharmaceutically acceptable carriers.

[0323] Another object of the present invention is to provide a medicine box comprising:

[0324] a) The compounds of the present invention or their pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites, or prodrugs, or the pharmaceutical compositions of the present invention; and

[0325] b) Optional packaging and / or instructions.

[0326] In this invention, "pharmaceuticalally acceptable carrier" refers to a diluent, excipient, vehicle, or medium that is administered co-administered with a therapeutic agent and is suitable, to the extent of reasonable medical judgment, for contact with human and / or other animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0327] The pharmaceutical compositions of the present invention can act systemically and / or locally. For this purpose, they can be administered via suitable routes, such as parenteral, local, intravenous, oral, subcutaneous, intra-arterial, intradermal, transdermal, rectal, intracranial, intraperitoneal, intranasal, intramuscular, or as inhalers.

[0328] For these routes of administration, the pharmaceutical compositions of the present invention can be administered in suitable dosage forms. These dosage forms include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, aqueous suspensions, injectable solutions, elixirs, syrups, etc.

[0329] The pharmaceutical compositions of the present invention can also be administered in the form of sterile injectable formulations, including sterile injectable water or oil suspensions or sterile injectable water or oil solutions.

[0330] The pharmaceutical compositions of the present invention may contain 0.01 mg to 1000 mg of the compounds of the present invention.

[0331] In some embodiments, the present invention provides a method for preparing a pharmaceutical composition or pharmaceutical formulation of the present invention, the method comprising combining a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, or prodrug with one or more pharmaceutically acceptable carriers.

[0332] The pharmaceutical compositions of the present invention may optionally be administered in combination with other agents that have at least some effect in treating various diseases. In some embodiments, the present invention provides a combination formulation of the compounds of the present invention with additional therapeutic agents for simultaneous, separate, or sequential treatment.

[0333] Treatment methods and uses

[0334] Another object of the present invention is to provide the compounds of the present invention or pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs thereof, or pharmaceutical compositions of the present invention, or kits of the present invention, for the prevention or treatment of diseases (particularly cancerous diseases) associated with Axl.

[0335] Another object of the present invention is to provide the use of the compounds of the present invention or pharmaceutically acceptable salts, stereoisomers, tautomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs thereof, or pharmaceutical compositions of the present invention, or cassettes of the present invention in the preparation of medicaments for the prevention or treatment of diseases (particularly tumor diseases) associated with Axl.

[0336] Another object of the present invention is to provide a method for preventing or treating diseases (particularly neoplastic diseases) associated with Axl, comprising administering to an individual in need a preventive or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug of the present invention, or a pharmaceutical composition of the present invention, or a cassette of the present invention.

[0337] As used in this article, the term "effective amount" refers to an amount sufficient to achieve the desired preventive or therapeutic effect, such as an amount that reduces one or more symptoms associated with the disease to be treated.

[0338] The dosing regimen can be adjusted to provide the optimal required response. For example, a single bolus injection can be administered, several fractions can be administered over time, or the dose can be proportionally reduced or increased as indicated by the urgency of the treatment situation. It should be noted that dosage values ​​can vary depending on the type and severity of the condition to be alleviated, and may include single or multiple doses. To further understand, for any given individual, the specific dosing regimen should be adjusted over time based on individual needs and the professional judgment of the person administering the composition or supervising its administration.

[0339] The amount of the compound of the present invention administered will depend on the individual being treated, the severity of the condition or illness, the rate of administration, the disposal of the compound, and the prescribing physician's judgment. Generally, the effective dose is from about 0.001 mg / kg body weight / day to about 10,000 mg / kg body weight / day. Where appropriate, the effective dose is from about 0.01 mg / kg body weight / day to about 1,000 mg / kg body weight / day. The dosage can be from about 0.01 to 1,000 mg / kg body weight daily, every two days, or every three days, typically from 0.1 to 500 mg / kg body weight. Exemplary treatment regimens are once or more daily, once or more weekly, or once or more monthly. The formulation is typically given multiple times, with intervals between single doses that can be daily, weekly, monthly, or annually. Alternatively, the formulation can be given in the form of a sustained-release formulation, in which case a lower dosing frequency is required. The dosage and frequency vary depending on the half-life of the formulation in the subject. They may also vary depending on whether it is a prophylactic or therapeutic treatment. In prophylactic use, relatively low doses are administered over a long period at relatively low frequency intervals. In therapeutic use, relatively high doses are sometimes required at relatively short intervals until disease progression is slowed or stopped, and preferably until the individual shows partial or complete improvement in disease symptoms, after which a prophylactic regimen can be given to the patient.

[0340] The amount of the compound of the present invention administered will depend on the individual being treated, the severity of the disease or condition, the rate of administration, the disposal of the compound, and the judgment of the prescribing physician.

[0341] As used in this invention, the term "treatment" aims to alleviate or eliminate a targeted disease state or symptom. A subject is successfully "treated" if, after receiving a therapeutic amount of a compound, its optical isomer, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, as described herein, one or more indications and symptoms exhibit an observable and / or detectable reduction or improvement. It should also be understood that treatment of the disease state or symptom includes not only complete treatment but also the achievement of some biologically or medically relevant outcome without achieving complete treatment.

[0342] "Treatment" means any administration of the compounds of the present invention, including:

[0343] (1) To prevent disease in animals that may be disease-prone but have not yet experienced or shown disease pathology or symptomology;

[0344] (2) Inhibit disease in animals that are experiencing or exhibiting disease pathology or symptomology (i.e., prevent further development of pathology and / or symptomology); or

[0345] (3) Improve disease (i.e., reverse pathology and / or symptomology) in animals that are experiencing or exhibiting disease pathology or symptomology.

[0346] As used herein, “individual” includes both human and non-human animals. Exemplary human individuals include human individuals suffering from a disease (such as the disease described herein) (referred to as patients) or normal individuals. In this invention, “non-human animals” includes all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock, and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

[0347] Example

[0348] To make the objectives and technical solutions of this invention clearer, the embodiments of this invention are described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of this invention. Unless otherwise specified, specific conditions in the examples are performed under conventional conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0349] In this application, when the chemical name and structural formula are inconsistent, the structural formula shall prevail, unless the chemical name rather than the structural formula can be inferred from the context to be correct.

[0350] The meanings of the abbreviations in the conventional synthesis methods and in the examples and intermediate synthesis examples are shown in the table below.

[0351]

[0352]

[0353] The structures of the compounds described in the following examples are obtained through... 1 Confirmed by H-NMR or MS. 1 The 1H-NMR measurements were performed using a Bruker 400MHz NMR spectrometer. The solvents used were CD3OD, CDCl3, or DMSO-d6, and the internal standard was TMS. All δ values ​​were expressed in ppm. The mass spectrometry (MS) measurements were performed using an Agilent (ESI) mass spectrometer, model Agilent 6120B.

[0354] Thin-layer chromatography (TLC) used Merck aluminum plates (20×20cm), and GF 254 (0.4-0.5mm) was used for TLC separation and purification.

[0355] The reaction was monitored using thin-layer chromatography (TLC) or LC-MS. The developing solvent systems used included dichloromethane and methanol, n-hexane and ethyl acetate, and petroleum ether and ethyl acetate. The volume ratio of the solvent was adjusted according to the polarity of the compound or by adding triethylamine, etc.

[0356] Column chromatography typically uses 200-300 mesh silica gel as the support. Eluent systems include dichloromethane and methanol, and petroleum ether and ethyl acetate. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of triethylamine can also be added for adjustment.

[0357] High-performance liquid chromatography (HPLC) was prepared using an Agilent 1260 instrument and a Waters XBridge PrepC column. 18 OBD (19mm×150mm×5.0μm); Column temperature: 25℃; Flow rate: 20.0mL / min; Detection wavelength: 214nm; Elution gradient: (0min: 10%A, 90%B; 16.0min: 90%A, 10%B); Mobile phase A: 100% acetonitrile; Mobile phase B: 0.05% ammonium bicarbonate aqueous solution.

[0358] Instrument model: Agilent 1260; Column: Waters SunFire Prep C18 OBD (19mm×150mm×5.0μm); Column temperature: 25℃; Flow rate: 20.0mL / min; Detection wavelength: 214nm; Elution gradient: (0min: 10%A, 90%B; 16.0min: 90%A, 10%B); Mobile phase A: 100% acetonitrile; Mobile phase B: 100% water, 0.05% formic acid.

[0359] Unless otherwise specified, the reaction temperature in the examples is room temperature (20°C to 30°C).

[0360] The reagents used in this invention were purchased from Acros Organics, Aldrich Chemical Company, Shanghai Teber Chemical Technology Co., Ltd., etc.

[0361] Example of intermediate preparation:

[0362] Intermediate Preparation Example 1: 5-(4-aminophenyl)-7-(1,1,1-trifluoropropane-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T1)

[0363]

[0364] Step 1: 1,1,1-Trifluoropropane-2-ylmethanesulfonate (T1-2)

[0365] Compound T1-1 (2.00 g, 17.54 mmol) was added to dichloromethane (20 mL), and the temperature was lowered to -10 °C. Then, triethylamine (3.55 g, 35.08 mmol) and methanesulfonyl chloride (4.02 g, 35.08 mol) were added, and the mixture was stirred continuously for 0.5 hours. Dilute hydrochloric acid (1 N) was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain crude compound T1-2 (3.00 g).

[0366] MS m / z (ESI): 193.2 [M+H] + .

[0367] Step 2: 5-Bromo-7-(1,1,1-trifluoropropane-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T1-3)

[0368] Compound T1-2 (1.24 g, 6.32 mmol) and 5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (1.00 g, 4.22 mmol) were added to N,N-dimethylformamide (20 mL), followed by potassium carbonate (1.18 g, 8.43 mmol). The temperature was raised to 110 °C, and the mixture was stirred continuously for 16 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give the title compound T1-3 (1.00 g).

[0369] MS m / z(ESI): 309.3 [M+H] + .

[0370] Step 3: 5-(4-aminophenyl)-7-(1,1,1-trifluoropropane-2-yl)-7H-pyrrolo[3,4-d]pyrimidin-4-ylamine (T1)

[0371] Compound T1-3 (888 mg, 2.58 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (685 mg, 3.10 mmol), sodium carbonate (828 mg, 7.74 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (572 mg, 0.77 mmol) were dissolved in a mixed solvent of 1,4-dioxane (10 mL) and water (1 mL), and reacted at 90 °C for 12 hours. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give the title compound T1 (600 mg).

[0372] MS m / z(ESI): 322.1 [M+H] + .

[0373] Intermediate Preparation Example 2: 3-(4-aminophenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T2)

[0374]

[0375] Step 1: 3-Bromo-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T2-1)

[0376] Compound T1-2 (2.2 g, 11.46 mmol) and 3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (1.00 g, 9.55 mmol) were added to N,N-dimethylformamide (20 mL), followed by potassium carbonate (2.64 g, 19.10 mmol). The temperature was raised to 110 °C, and the mixture was stirred continuously for 16 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give the title compound T2-1 (1.51 g).

[0377] MS m / z (ESI): 310.2 [M+H] + .

[0378] Step 3: 3-(4-aminophenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T2)

[0379] Compound T2-1 (500 mg, 1.62 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (429 mg, 1.94 mmol), sodium carbonate (448 mg, 3.24 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (119 mg, 0.16 mmol) were dissolved in a mixed solvent of 1,4-dioxane (10 mL) and water (1 mL), and reacted at 90 °C for 12 hours. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give intermediate T2 (350 mg).

[0380] Intermediate Preparation Example 3: 5-(4-aminophenyl)-7-(2,2-difluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T3)

[0381]

[0382] Step 1: 2,2-Difluoroethyl methanesulfonate (T3-2)

[0383] Compound T3-1 (2.00 g, 24.39 mmol) was added to dichloromethane (20 mL), and the temperature was lowered to -10 °C. Then, triethylamine (4.94 g, 48.78 mmol) and methanesulfonyl chloride (5.59 g, 48.78 mol) were added, and the mixture was stirred continuously for 0.5 hours. Dilute hydrochloric acid (1 N) was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give crude compound T3-2 (2.60 g).

[0384] MS m / z (ESI): 161.1 [M+H] + .

[0385] Step 2: 5-Bromo-7-(2,2-difluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T3-3)

[0386] Compound T3-2 (1.24 g, 7.75 mmol) and 5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (1.10 g, 5.17 mmol) were added to N,N-dimethylformamide (20 mL), followed by potassium carbonate (1.43 g, 10.33 mmol). The temperature was raised to 110 °C and stirred continuously for 16 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate. The concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give the title compound T1-3 (0.95 g).

[0387] MS m / z(ESI): 277.1 [M+H] + .

[0388] Step 3: 5-(4-aminophenyl)-7-(2,2-difluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T3)

[0389] Compound T3-3 (800 mg, 2.90 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (769 mg, 3.48 mmol), sodium carbonate (931 mg, 8.70 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (212 mg, 0.29 mmol) were dissolved in a mixed solvent of 1,4-dioxane (10 mL) and water (1 mL), and reacted at 90 °C for 12 hours. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give the title compound T3 (560 mg).

[0390] MS m / z(ESI): 290.2 [M+H] + .

[0391] Intermediate Preparation Example 4: 5-(4-aminophenyl)-7-isobutyl-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T4)

[0392]

[0393] Step 1: Isobutyl mesylate (T4-2)

[0394] Compound T4-1 (2.00 g, 27.00 mmol) was added to dichloromethane (20 mL), and the temperature was lowered to -10 °C. Then, triethylamine (5.47 g, 54.00 mmol) and methanesulfonyl chloride (6.19 g, 54.00 mol) were added, and the mixture was stirred continuously for 0.5 hours. Dilute hydrochloric acid (1 N) was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain crude compound T4-2 (2.40 g).

[0395] MS m / z (ESI): 153.1 [M+H] + .

[0396] Step 2: 5-Bromo-7-isobutyl-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T4-3)

[0397] Compound T4-2 (1.24 g, 8.16 mmol) and 5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (1.16 g, 5.44 mmol) were added to N,N-dimethylformamide (20 mL), followed by potassium carbonate (1.51 g, 10.87 mmol). The temperature was raised to 110 °C and stirred continuously for 16 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate. The concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give the title compound T4-3 (1.02 g).

[0398] MS m / z(ESI): 269.1 [M+H] + .

[0399] Step 3: 5-(4-aminophenyl)-7-isobutyl-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T4)

[0400] Compound T4-3 (700 mg, 2.61 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (692 mg, 3.13 mmol), sodium carbonate (838 mg, 7.83 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (191 mg, 0.26 mmol) were dissolved in a mixed solvent of 1,4-dioxane (10 mL) and water (1 mL), and reacted at 90 °C for 12 hours. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give the title compound T4 (480 mg).

[0401] MS m / z(ESI): 282.2 [M+H] + .

[0402] Intermediate Preparation Example 5: 5-(4-aminophenyl)-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T5)

[0403]

[0404] Step 1: 5-Bromo-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T5-2)

[0405] Compound T5-1 (1.4 g, 6.4 mmol), 2-iodo-1,1,1-trifluoroethane (4.27 g, 19.32 mmol), and potassium carbonate (2.7 g, 19.32 mmol) were added to N,N-dimethylformamide (15 mL), and the mixture was reacted at 80 °C for 16 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give the title compound T5-2 (350 mg).

[0406] MS m / z(ESI): 295.2 [M+H] + .

[0407] Step 2: 5-(4-aminophenyl)-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T5)

[0408] Compound T5-2 (200 mg, 0.66 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (162 mg, 0.73 mmol), sodium carbonate (140 mg, 1.31 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (50 mg, 0.066 mmol) were added to dioxane / water (2 mL / 0.4 mL) and reacted at 90 °C for 3 hours under nitrogen. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give the title compound T5 (150 mg).

[0409] MS m / z (ESI): 308.2 [M+H] + .

[0410] Intermediate Preparation Example 6: 3-(4-aminophenyl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T6)

[0411]

[0412] Step 1: 3-Bromo-1-tetrahydropyran-4-yl-pyrazolo[3,4-d]pyrimidin-4-ylamine (T6-2)

[0413] Compound T6-1 (1 g, 4.58 mmol) was added to tetrahydrofuran (15 mL), followed by tetrahydropyran-4-ol (937 mg, 9 mmol) and triphenylphosphine (2.45 g, 9.16 mmol). Diethyl azodicarbonate (1.63 g, 9.16 mmol) was added dropwise under nitrogen atmosphere, and the reaction was carried out at 25 °C for 16 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give the crude product, compound T6-2 (1 g).

[0414] MS m / z(ESI): 298.1 [M+H] + .

[0415] Step 2: 3-(4-aminophenyl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T6)

[0416] Compound T6-2 (1 g, 3.35 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (734 mg, 3.35 mmol), sodium carbonate (710 mg, 6.7 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (277 mg, 0.34 mmol) were added to dioxane / water (5 mL / 1 mL) and reacted at 110 °C for 5 hours under nitrogen. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give the title compound T6 (450 mg).

[0417] MS m / z(ESI): 311.2 [M+H] + .

[0418] Intermediate Preparation Example 7: 5-(4-aminophenyl)-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T7)

[0419]

[0420] Step 1: 5-Bromo-4-chloro-7-tetrahydropyran-4-yl-pyrrolo[2,3-d]pyrimidine (T7-2)

[0421] Compound T7-1 (500 mg, 2.11 mmol) was added to 1,4-dioxane (5 mL), followed by tetrahydropyran-4-ol (330 mg, 3.16 mmol) and triphenylphosphine (1.15 g, 4.30 mmol). Diethyl azodicarbonate (765 mg, 4.30 mmol) was added dropwise under nitrogen atmosphere, and the mixture was microwaved at 85 °C for 1 hour. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give the title compound T7-2 (450 mg).

[0422] MS m / z (ESI): 317.1 [M+H] + .

[0423] Step 2: 5-Bromo-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T7-3)

[0424] Compound T7-2 (350 mg, 1.05 mmol) was added to dioxane (2 ml), followed by ammonia (2 ml). The mixture was then microwaved at 90 °C for 2 hours. After the reaction was complete, excess solvent was evaporated under reduced pressure to obtain the crude title compound T7-3 (300 mg).

[0425] MS m / z(ESI): 297.0 [M+H] + .

[0426] Step 3: 5-(4-aminophenyl)-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T7)

[0427] Compound T7-3 (300 mg, 960 μmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (425 mg, 1.92 mmol), cesium carbonate (645 mg, 1.92 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (90 mg, 96 μmol) were added to dioxane / water (5 mL / 1 mL), and reacted at 90 °C for 16 hours under nitrogen. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give the title compound T7 (240 mg).

[0428] MS m / z (ESI): 310.3 [M+H] + .

[0429] Intermediate Preparation Example 8: 3-(4-aminophenyl)-1-propyl-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T8)

[0430]

[0431] Step 1: 3-Bromo-1-propyl-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T8-2)

[0432] Compound T6-1 (816 g, 3.74 mmol), 1-iodopropane (642 mg, 3.74 mmol), and potassium carbonate (1.58 g, 11.21 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was heated to 80 °C and reacted for 12 hours. After the reaction was complete, the mixture was cooled to room temperature, water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, and concentrated to give the title compound T8-1 (410 mg).

[0433] MS m / z(ESI): 258.1 [M+H] + .

[0434] Step 2: 3-(4-aminophenyl)-1-propyl-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T8)

[0435] Compound T8-1 (410 mg, 1.44 mmol), 4-aminophenylboronic acid pinacol ester (322 mg, 1.44 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (108 mg, 0.14 mmol), and cesium carbonate (948 mg, 2.88 mmol) were dissolved in dioxane (15 mL) and water (3 mL). The mixture was heated to 90 °C under nitrogen protection and reacted for 14 hours. After the reaction was completed, the mixture was cooled to room temperature, concentrated, and separated by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the title compound T8 (220 mg).

[0436] MS m / z(ESI): 269.2 [M+H] + .

[0437] Intermediate Preparation Example 9: 3-(4-aminophenyl)-1-(oxacyclobut-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T9)

[0438]

[0439] Step 1: Cyclobutyl mesylate (T9-2)

[0440] Compound T9-1 (1 g, 13.5 mmol) was dissolved in dichloromethane (10 mL), and triethylamine (2.7 g, 27 mmol) was added. Methanesulfonyl chloride (2.32 g, 20.3 mmol) was then added at 0 °C, and the reaction was allowed to proceed for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate and concentrated to give the title compound T9-2 (2 g).

[0441] Step 2: 3-Bromo-1-(oxetane-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T9-3)

[0442] Compounds T9-2 (748 mg, 4.67 mmol) and T6-1 (500 mg, 2.34 mmol) were dissolved in N,N-dimethylformamide (5 mL), and potassium carbonate (969 mg, 7 mmol) was added. The mixture was stirred at 80 °C for 16 hours. Water was added to the reaction solution, followed by extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, concentrating, and then slurrying with dichloromethane to obtain the title compound T9-3 (500 mg).

[0443] MS m / z(ESI): 270.1 [M+H] + .

[0444] Step 3: 3-(4-aminophenyl)-1-(oxacyclobut-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T9)

[0445] Compound T9-3 (300 mg, 1.06 mmol) was dissolved in dioxane (5 mL) and water (1 mL). Sodium carbonate (224 mg, 2.11 mmol) was added, along with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)aniline (347 mg, 1.58 mmol) and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane dichloride complex (172 mg, 211 μmol). The mixture was stirred at 100 °C for 16 hours under nitrogen protection. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed three times with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was subjected to silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give intermediate T9 (300 mg).

[0446] MS m / z(ESI): 283.1 [M+H] + .

[0447] Intermediate Preparation Example 10: 3-(4-aminophenyl)-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T10)

[0448]

[0449] Step 1: Tetrahydrofuran-3-ylmethanesulfonate (T10-2)

[0450] Compound T10-1 (1.19 g, 13.5 mmol) was added to dichloromethane (10 mL), followed by triethylamine (2.7 g, 27 mmol). Methanesulfonyl chloride (2.32 g, 20.3 mmol) was then added at 0 °C, and the reaction was allowed to proceed for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate and concentrated to give the title compound T10-2 (2.1 g).

[0451] Step 2: 3-Bromo-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T10-3)

[0452] Compound T10-2 (878 mg, 4.67 mmol) and compound T6-1 (500 mg, 2.34 mmol) were added to N,N-dimethylformamide (5 mL), followed by potassium carbonate (969 mg, 7 mmol). The mixture was stirred at 80 °C for 16 hours. Water was added to the reaction mixture, followed by extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, concentrating, and then slurrying with dichloromethane to obtain the title compound T10-3 (510 mg).

[0453] MS m / z(ESI): 270.1 [M+H] + .

[0454] Step 3: 3-(4-aminophenyl)-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T10)

[0455] Compound T10-3 (300 mg, 1.06 mmol) was added to dioxane (5 mL) and water (1 mL), followed by sodium carbonate (224 mg, 2.11 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)aniline (347 mg, 1.58 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane dichloride complex (172 mg, 211 μmol). The mixture was stirred at 100 °C for 16 hours under nitrogen protection. Water was added to the reaction mixture, and the solution was extracted with ethyl acetate. The organic phases were combined, washed three times with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The concentrated residue was subjected to silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give intermediate T10 (300 mg).

[0456] MS m / z(ESI): 283.1 [M+H]+ .

[0457] Intermediate Preparation Example 11: 3-(4-amino-3-fluorophenyl)-1-(oxacyclobut-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T11)

[0458]

[0459] Compound T9-3 (300 mg, 1.00 mmol), 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (242 mg, 1.00 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (75 mg, 0.1 mmol), and cesium carbonate (658 mg, 2 mmol) were added to dioxane (20 mL) and water (4 mL). The mixture was heated to 90 °C under nitrogen protection and reacted for 14 hours. After the reaction was complete, the mixture was cooled to room temperature, concentrated, and separated by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the title compound T11 (240 mg).

[0460] MS m / z (ESI): 301.1 [M+H] + .

[0461] Intermediate Preparation Example 12: 2-(5-methylpyridin-2-yl)-3-oxo-6-((tetrahydro-2H-pyran-4-yl)methyl)-2,3-dihydropyridazine-4-carboxylic acid (T12)

[0462]

[0463] Step 1: 3-(4-hydroxytetrahydro-2H-pyran-4-yl)-1,1-dimethoxypropane-2-one (T12-2)

[0464] Diisopropylaminolithium (2M, 55.02 mL) was weighed and added to tetrahydrofuran (150 mL). T12-1 (10 g, 84.7 mmol) was added at -70 °C, and the mixture was stirred at -70 °C for 1 hour. Tetrahydropyran-4-one (8.5 g, 84.7 mmol) was then added, and the mixture was stirred at -70 °C for 2 hours. A saturated ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give the title compound T12-2 (3 g).

[0465] Step 2: 1,1-Dimethoxy-3-(tetrahydro-2H-pyran-4-ylidene)prop-2-one (T12-3)

[0466] Compound T12-2 (3.5 g, 15.2 mmol) was added to dichloromethane (40 mL), followed by the addition of pyridine (6.03 g, 76.2 mmol) and thionyl chloride (5.44 g, 45.7 mmol) at 0 °C. The mixture was stirred at 0 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined, the pH was adjusted to alkaline with triethylamine, washed with saturated brine, and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give the title compound T12-3 (2.5 g).

[0467] Step 3: 1,1-Dimethoxy-3-(tetrahydro-2H-pyran-4-yl)prop-2-one (T12-4)

[0468] Compound T12-3 (2.5 g, 11.9 mmol) was added to methanol (50 mL), followed by palladium / carbon (1 g, 10% pure). The reaction was carried out under hydrogen (15 PSI) protection for 16 hours. The reaction solution was filtered directly, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give the title compound T12-4 (1 g).

[0469] Step 4: (Z)-2-(2-(1,1-dimethoxy-3-(tetrahydro-2H-pyran-4-yl)propyl-2-yl)hydrazino)-5-methylpyridine (T12-5)

[0470] (5-Methyl-2-pyridyl)hydrazine (600 mg, 4.9 mmol) was weighed and added to methanol (5 mL), followed by compound T12-4 (1.04 g, 4.9 mmol). The mixture was reacted at 65 °C for 3 hours. The reaction solution was directly concentrated, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give the title compound T12-5 (1 g).

[0471] MS m / z (ESI): 308.1 [M+H] + .

[0472] Step 5: (Z)-2-(2-(5-methylpyridin-2-yl)hydrazone)-3-(tetrahydro-2H-pyran-4-yl)propionaldehyde (T12-6)

[0473] Compound T12-5 (900 mg, 2.64 mmol) was weighed and added to dilute hydrochloric acid (6 mL, 2N), and reacted at 25 °C for 16 hours. A saturated potassium carbonate solution was added to the reaction mixture to bring the pH to 10, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give the title compound T12-6 (600 mg).

[0474] MS m / z(ESI): 262.1 [M+H] + .

[0475] Step Six: ((Z)-2,2-dimethyl-5-(2-(2-(5-methylpyridin-2-yl)hydrazone)-3-(tetrahydro-2H-pyran-4-yl)propylidene)-1,3-dioxane-4,6-dione (T12-7)

[0476] Compound T12-6 (600 mg, 2.07 mmol) was weighed and added to toluene (20 mL), followed by 2,2-dimethyl-1,3-dioxane-4,6-dione (298 mg, 2.07 mmol), acetic acid (0.5 mL), and tetrahydropyrrole (0.5 mL). The reaction mixture was reacted at 20 °C for 16 hours. The reaction solution was filtered directly, and the filter cake was washed with toluene to obtain the title compound T12-7 (800 mg).

[0477] MS m / z(ESI): 330.2 [M+H-58] + .

[0478] Step 7: 2-(5-methylpyridin-2-yl)-3-oxo-6-((tetrahydro-2H-pyran-4-yl)methyl)-2,3-dihydropyridazine-4-carboxylic acid (T12)

[0479] Compound T12-7 (600 mg, 1.39 mmol) was weighed and dissolved in methanol (90 mL). Sodium methoxide (90 mg, 1.67 mmol) was added, and the mixture was reacted at 65 °C for 16 hours. The reaction solution was concentrated to remove methanol, and the pH was adjusted to 6 with dilute hydrochloric acid (1 N). The solution was then extracted with a dichloromethane / methanol mixture. The extract was dried over anhydrous sodium sulfate and concentrated to give intermediate T12 (240 mg).

[0480] MS m / z (ESI): 330.2 [M+H] + .

[0481] Intermediate Preparation Example 13: 6-Isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13)

[0482]

[0483] Step 1: 1,1-Dimethoxy-3-methylbutane-2-one (T13-2)

[0484] Compound T13-1 (50 g, 373 mmol) and hydroxylamine hydrochloride (45.5 g, 466 mmol) were weighed and added to tetrahydrofuran (500 mL). Isopropyl magnesium bromide (1 M, 931.93 mL) was added at -20 °C, and the temperature was maintained between -15 °C and 20 °C, with stirring for 2 hours. A saturated aqueous solution of ammonium chloride was added, and the mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give the title compound T13-2 (16 g).

[0485] Step 2: (Z)-2-(2-(1,1-dimethoxy-3-methylbutane-2-yl)hydrazino)-5-methylpyridine (T13-3)

[0486] (5-methyl-2-pyridyl)hydrazine (6 g, 43.9 mmol) was weighed and dissolved in methanol (200 mL). Compound T13-2 (10.7 g, 65.8 mmol) was added, and the mixture was reacted at 65 °C for 2 hours. The concentrated residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give the title compound T13-3 (11 g).

[0487] MS m / z(ESI): 252.1 [M+H] + .

[0488] Step 3: (Z)-3-methyl-2-(2-(5-methylpyridin-2-yl)hydrazone)butyraldehyde (T13-4)

[0489] Compound T13-3 (11 g, 41.6 mmol) was weighed and added to dilute hydrochloric acid (170 mL, 2 M), and reacted at 25 °C for 16 hours. Saturated potassium carbonate solution was added to pH 10, and the mixture was extracted with a dichloromethane / methanol mixture. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give the title compound T13-4 (8.5 g).

[0490] MS m / z(ESI): 206.1 [M+H] + .

[0491] Step 4: (Z)-2,2-dimethyl-5-{3-methyl-2-[(5-methylpyridin-2-yl)hydrazone]butylene}-[1,3]dioxane-4,6-dione (T13-5)

[0492] Compound T13-4 (8 g, 37 mmol) was weighed and added to toluene (160 mL), followed by 2,2-dimethyl-1,3-dioxane-4,6-dione (6.40 g, 44.4 mmol), acetic acid (8 mL), and tetrahydropyrrole (8 mL). The reaction was carried out at 25 °C for 16 hours, resulting in the formation of a solid. The reaction solution was directly filtered to obtain the title compound T13-5 (12 g).

[0493] MS m / z (ESI): 274.2 [M+H-58] + .

[0494] Step 5: 6-Isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13)

[0495] T13-5 (16 g, 45.9 mmol) was added to methanol (160 mL), followed by sodium methoxide (4.1 g, 68.8 mmol), and the reaction was carried out at 65 °C for 16 hours. The reaction solution was concentrated to remove methanol, the pH was adjusted to 6 with dilute hydrochloric acid (1 N), and then extracted with a dichloromethane / methanol mixture. The solution was dried over anhydrous sodium sulfate and concentrated to obtain intermediate T13 (10 g).

[0496] MS m / z(ESI): 274.2 [M+H] + .

[0497] Intermediate Preparation Example 14: 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14)

[0498]

[0499] Step 1: 1-Bromo-3-methyl-butane-2-one (T14-2)

[0500] Compound T14-1 (100 g, 1.16 mol) was weighed and added to methanol (1 L). Liquid bromine (186 g, 1.16 mol) was added at 0 °C, and the mixture was stirred at 0 °C for 2 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate, washed with saturated brine, washed with sodium bicarbonate aqueous solution until neutral, dried over anhydrous sodium sulfate, and concentrated to obtain the title compound T14-2 (150 g).

[0501] Step 2: Dimethyl 2-(3-methyl-2-oxobutyl)malonate (T14-3)

[0502] Dimethyl malonate (123 g, 933 mmol) was weighed and added to tetrahydrofuran (2.7 L). Sodium hydride (34 g, 855 mmol, 60% pure) was added at 0 °C, and the mixture was stirred for half an hour. Then, compound T14-2 (135 g, 777.14 mmol) was added, and the mixture was stirred at 0 °C for 2 hours. Dilute hydrochloric acid (1 N) was added to the reaction solution until the pH was acidic. The solution was extracted with ethyl acetate and dried over anhydrous sodium sulfate. The concentrated residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give the title compound T14-3 (160 g).

[0503] MS m / z(ESI): 217.1 [M+H] + .

[0504] Step 3: Methyl 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3,4,5-tetrahydropyridazine-4-carboxylate (T14-4)

[0505] Compound T13-3 (83 g, 365 mmol) was added to acetic acid (900 mL), along with (4-fluorophenyl)hydrazine hydrochloride (49 g, 304 mmol) and sodium acetate (75 g, 912 mmol). The mixture was stirred at 50 °C for 16 hours. The reaction solution was poured into water, extracted with ethyl acetate, washed with saturated brine, washed with sodium bicarbonate solution until alkaline, dried over anhydrous sodium sulfate, and concentrated to give the title compound T14-4 (85 g).

[0506] MS m / z(ESI): 293.2 [M+H] + .

[0507] Step 4: Methyl 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylate (T14-5)

[0508] Compound T13-4 (30 g, 97.5 mmol) was dissolved in acetonitrile (300 mL), and copper chloride (20 g, 146 mmol) was added. The mixture was stirred at 70 °C for 40 hours. The reaction solution was filtered directly, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give the title compound T13-5 (20 g).

[0509] MS m / z(ESI): 291.2 [M+H] + .

[0510] Step 5: 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14)

[0511] Compound T13-5 (170 g, 556 mmol) was added to water (350 mL) and methanol (1.7 L), and lithium hydroxide monohydrate (70 g, 1.67 mol) was added. The reaction mixture was reacted at 25 °C for 16 hours. The reaction solution was concentrated, and water and dilute hydrochloric acid (1 N) were added to adjust the pH to acidic. The mixture was then extracted with a dichloromethane / methanol mixed solvent and concentrated to give the title compound T14 (80 g).

[0512] MS m / z(ESI): 277.1 [M+H] + .

[0513] Intermediate Preparation Example 15: 2-(5-Fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15)

[0514]

[0515] Step 1: Methyl 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3,4,5-tetrahydropyridazine-4-carboxylate (T15-1)

[0516] Compound T14-3 (40 g, 185 mmol) was added to acetic acid (500 mL), along with 5-fluoro-2-hydrazinopyridine hydrochloride (25 g, 154 mmol) and sodium acetate (38 g, 462 mmol). The mixture was stirred at 50 °C for 16 hours. The reaction solution was poured into water, extracted with ethyl acetate, washed with saturated brine, washed with sodium bicarbonate solution until alkaline, dried over anhydrous sodium sulfate, and concentrated to give the title compound T15-1 (31 g).

[0517] MS m / z(ESI): 294.1 [M+H] + .

[0518] Step 2: Methyl 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylate (T15-2)

[0519] Compound T15-1 (15 g, 48.75 mmol) was dissolved in acetonitrile (200 mL), and copper chloride (10 g, 73 mmol) was added. The mixture was stirred at 70 °C for 40 hours. The reaction solution was filtered directly, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give the title compound T15-2 (10 g).

[0520] MS m / z(ESI): 292.1 [M+H] + .

[0521] Step 3: 2-(5-Fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15)

[0522] Compound T15-2 (170 g, 556 mmol) was added to water (350 mL) and methanol (1.7 L), followed by the addition of lithium hydroxide monohydrate (70 g, 1.67 mol). The reaction mixture was reacted at 25 °C for 16 hours. The reaction solution was concentrated, and water and dilute hydrochloric acid (1 N) were added to adjust the pH to acidic. The mixture was then extracted with a dichloromethane / methanol mixed solvent and concentrated to obtain the title compound T15 (80 g).

[0523] MS m / z(ESI): 278.1 [M+H] + .

[0524] Intermediate Preparation Example 16: 6-Isopropyl-3-oxo-2-[5-(trifluoromethyl)-2-pyridyl]-2,3-dihydropyridazine-4-carboxylic acid (T16)

[0525]

[0526] Step 1: Dimethyl 2-[(2E)-3-methyl-2-[[5-(trifluoromethyl)-2-pyridyl]hydrazone]butyl]malonate (T16-1)

[0527] Compound T14-3 (9.3 g, 40.7 mmol) was added to methanol (30 mL), along with 2-hydrazino-5-(trifluoromethyl)pyridine (6 g, 33.9 mmol) and a catalytic amount of acetic acid. The mixture was stirred at 70 °C for 16 hours. The reaction solution was then concentrated to remove methanol, yielding the title compound T16-1 (12 g).

[0528] MS m / z (ESI): 376.2 [M+H] + .

[0529] Step 2: Methyl 6-isopropyl-3-oxo-2-[5-(trifluoromethyl)-2-pyridyl]-2,3,4,5-tetrahydropyridazine-4-carboxylate (T16-2)

[0530] Compound T16-1 (4.2 g, 11.3 mmol) and sodium acetate (4.6 g, 33.9 mmol) were added to acetic acid (30 mL) and reacted at 80 °C for 16 hours. The reaction solution was poured into water, extracted with ethyl acetate, washed three times with water, and then washed with sodium carbonate solution until the pH was alkaline. The solution was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give the title compound T16-2 (1.5 g).

[0531] MS m / z(ESI): 244.2 [M+H] + .

[0532] Step 3: Methyl 6-isopropyl-3-oxo-2-[5-(trifluoromethyl)-2-pyridyl]-2,3-dihydropyridazine-4-carboxylate (T16-3)

[0533] Compound T16-2 (1.3 g, 3.4 mmol) was added to methanol (20 mL), followed by palladium / carbon (500 mg), and reacted at 50 °C for 16 hours in air. The reaction was directly filtered, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give the title compound T16-3 (1 g).

[0534] MS m / z(ESI): 242.2 [M+H] + .

[0535] Step 4: 6-Isopropyl-3-oxo-2-[5-(trifluoromethyl)-2-pyridyl]-2,3-dihydropyridazine-4-carboxylic acid (T16)

[0536] Compound T16-3 (1 g, 2.78 mmol) was added to water (2 mL) and methanol (10 mL), followed by lithium hydroxide monohydrate (200 mg, 8.35 mmol). The reaction mixture was reacted at 25 °C for 4 hours. The methanol was removed by concentration, and the reaction solution was concentrated. Water and dilute hydrochloric acid (1 N) were added to adjust the pH to acidic. The mixture was then extracted with a dichloromethane / methanol mixed solvent and concentrated to obtain intermediate T16 (900 mg).

[0537] MS m / z(ESI): 328.1 [M+H] + .

[0538] Intermediate Preparation Example 17: 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17)

[0539]

[0540] Step 1: Methyl 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3,4,5-tetrahydropyridazine-4-carboxylate (T17-1)

[0541] Compound T14-3 (3.00 g, 25.49 mmol) and 4-chlorophenylhydrazine hydrochloride (4.32 g, 30.47 mmol) were added to acetic acid (50 mL), followed by sodium acetate (6.94 g, 50.98 mmol). The temperature was raised to 80 °C, and the mixture was stirred continuously for 16 hours. The mixture was washed with a saturated sodium carbonate solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give crude compound T17-1 (3.20 g).

[0542] MS m / z(ESI): 309.1 [M+H] + .

[0543] Step 2: Methyl 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylate (T17-2)

[0544] Compound T17-1 (2.64 g, 6.82 mmol) was added to methanol (40 mL), followed by palladium / carbon (4.14 g, 10%). The mixture was heated to 50 °C and exposed to air with continuous stirring for 16 hours. The residue after filtration and concentration was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give the title compound T17-2 (1.00 g).

[0545] MS m / z (ESI): 307.3 [M+H] + .

[0546] Step 3: 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17)

[0547] Compound T17-2 (1.12 g, 3.26 mmol) was added to methanol (50 mL), followed by lithium hydroxide hydrate (160 mg, 3.72 mmol), and the reaction was carried out at 25 °C for 12 hours. The pH was adjusted with dilute hydrochloric acid (1 N) to precipitate a solid, which was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give intermediate T17 (600 mg).

[0548] MS m / z(ESI): 293.1 [M+H] + .

[0549] Intermediate Preparation Example 18: 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18)

[0550]

[0551] Step 1: Methyl 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3,4,5-tetrahydropyridazine-4-carboxylate (T18-1)

[0552] Compound T14-3 (6.10 g, 25.49 mmol) and 5-chloro-2-hydrazinopyridine hydrochloride (5.60 g, 30.47 mmol) were added to acetic acid (50 mL), followed by sodium acetate (5.82 g, 70.22 mmol). The temperature was raised to 80 °C, and the mixture was stirred continuously for 16 hours. The mixture was washed with a saturated sodium carbonate solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give crude compound T18-1 (6.00 g).

[0553] MS m / z (ESI): 310.2 [M+H] + .

[0554] Step 2: Methyl 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylate (T18-2)

[0555] Compound T18-1 (2.64 g, 6.82 mmol) was added to methanol (40 mL), followed by palladium / carbon (4.14 g, 10%). The mixture was heated to 50 °C and exposed to air with continuous stirring for 16 hours. The residue after filtration and concentration was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1-2 / 1) to give the title compound T18-2 (1.00 g).

[0556] MS m / z (ESI): 308.3 [M+H] + .

[0557] Step 3: 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18)

[0558] Compound T18-2 (1.12 g, 3.26 mmol) was added to methanol (50 mL), followed by lithium hydroxide hydrate (160 mg, 3.72 mmol), and the reaction was carried out at 25 °C for 12 hours. The pH was adjusted with dilute hydrochloric acid (1 N) to precipitate a solid, which was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give intermediate T18 (600 mg).

[0559] MS m / z(ESI): 294.1 [M+H] + .

[0560] Intermediate Preparation Example 19: 6-Isopropyl-3-oxo-2-(4-(trifluoromethyl)phenyl)-2,3-dihydropyridazine-4-carboxylic acid (T19)

[0561]

[0562] Step 1: Methyl 6-isopropyl-3-oxo-2-(4-(trifluoromethyl)phenyl)-2,3,4,5-tetrahydropyridazine-4-carboxylate (T19-1)

[0563] Compound T14-3 (1 g, 4.39 mmol) and sodium acetate (2.39 g, 17.57 mmol) were added to acetic acid (10 mL), followed by p-trifluoromethylphenylhydrazine hydrochloride (1.42 g, 6.59 mmol). The mixture was stirred at room temperature for 1 hour, then heated to 50 °C and reacted for 8 hours. The solution was evaporated to dryness, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give the title compound T19-1 (750 mg).

[0564] MS m / z(ESI): 343.1 [M+H] + .

[0565] Step 2: Methyl 6-isopropyl-3-oxo-2-(4-(trifluoromethyl)phenyl)-2,3-dihydropyridazine-4-carboxylate (T19-2)

[0566] Compound T19-1 (450 mg, 1.26 mmol) was added to acetonitrile (10 mL), followed by copper chloride (259 mg, 1.89 mmol). The mixture was reacted at 85 °C for 2 hours. The solution was evaporated to dryness, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate. The concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give the title compound T19-2 (300 mg).

[0567] MS m / z (ESI): 341.2 [M+H] + .

[0568] Step 3: 6-Isopropyl-3-oxo-2-(4-(trifluoromethyl)phenyl)-2,3-dihydropyridazine-4-carboxylic acid (T19)

[0569] Compound T19-2 (200 mg, 0.53 mmol) was added to methanol (5 mL), followed by lithium hydroxide (26 mg, 1.06 mmol). The mixture was reacted at 25 °C for 4 hours. The reaction solution was then directly evaporated to dryness and concentrated to obtain the title compound T19 (135 mg).

[0570] MS m / z (ESI): 327.1 [M+H] + .

[0571] Intermediate Preparation Example 20: 3-(4-aminophenyl)-1-isobutyl-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T20)

[0572]

[0573] Step 1: 3-Bromo-1-isobutyl-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T20-1)

[0574] Compound T6-1 (1.03 g, 4.67 mmol), 1-iodo-2-methylpropane (700 mg, 3.74 mmol), and potassium carbonate (1.32 g, 9.34 mmol) were dissolved in N,N-dimethylformamide (20 mL) and reacted at 80 °C for 8 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give the crude product, compound T20-1 (1.0 g).

[0575] MS m / z(ESI): 270.1 [M+H] + .

[0576] Step 2: 3-(4-aminophenyl)-1-isobutyl-1H-pyrazolo[3,4-d]pyrimidine-4-ylamine (T20)

[0577] Compound T20-1 (500 mg, 1.11 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (250 mg, 1.11 mmol), cesium carbonate (730 mg, 2.22 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (80 mg, 0.11 mmol) were dissolved in dioxane / water (5 mL / 1 mL) and reacted under nitrogen atmosphere at 90 °C with microwave for 1.5 h. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give the title compound T20 (200 mg).

[0578] MS m / z(ESI): 283.3 [M+H] + .

[0579] Intermediate Preparation Example 21: 3-(4-aminophenyl)-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T21)

[0580]

[0581] Step 1: 3-Bromo-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T21-1)

[0582] Compound T6-1 (510 mg, 2.34 mmol), 2,2-difluoroethyl methanesulfonate (780 mg, 4.67 mmol), and potassium carbonate (990 mg, 7.01 mmol) were dissolved in N,N-dimethylformamide (10 mL) and reacted at 85 °C for 8 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give the crude product, compound T21-1 (600 mg).

[0583] MS m / z(ESI): 278.1 [M+H] + .

[0584] Step 2: 3-(4-aminophenyl)-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-d]pyrimidine-4-ylamine (T21)

[0585] Compound T21-1 (270 mg, 0.88 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (200 mg, 0.88 mmol), cesium carbonate (580 mg, 1.78 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (66 mg, 0.088 mmol) were dissolved in dioxane / water (5 mL / 1 mL) and reacted under nitrogen atmosphere at 90 °C with microwave for 1.5 h. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give the title compound T21 (250 mg).

[0586] MS m / z(ESI): 291.1 [M+H] + .

[0587] Intermediate Preparation Example 22: 3-(4-aminophenyl)-1-(2-fluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T22)

[0588]

[0589] Step 1: 3-Bromo-1-(2-fluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T22-1)

[0590] Compound T6-1 (300 mg, 1.37 mmol), 2-fluoroethyl methanesulfonate (240 mg, 1.37 mmol), and potassium carbonate (580 mg, 4.12 mmol) were dissolved in N,N-dimethylformamide (10 mL) and reacted in a microwave oven at 90 °C for 1 hour. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give the crude product, compound T22-1 (300 mg).

[0591] MS m / z(ESI): 260.1 [M+H] + .

[0592] Step 2: 3-(4-aminophenyl)-1-(2-fluoroethyl)-1H-pyrazolo[3,4-d]pyrimidine-4-ylamine (T22)

[0593] Compound T22-1 (300 mg, 0.99 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (220 mg, 0.99 mmol), cesium carbonate (660 mg, 1.98 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (74 mg, 0.099 mmol) were dissolved in dioxane / water (5 mL / 1 mL) and reacted at 85 °C for 8 hours under nitrogen. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give the title compound T22 (200 mg).

[0594] MS m / z(ESI): 273.2 [M+H] + .

[0595] Intermediate Preparation Example 23: 3-(4-aminophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T23)

[0596]

[0597] Step 1: 3-Bromo-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T23-1)

[0598] Compound T6-1 (510 mg, 2.34 mmol), 2-bromopropane (323 mg, 2.57 mmol), and potassium carbonate (1.30 g, 9.33 mmol) were dissolved in N,N-dimethylformamide (8 mL), heated to 80 °C, and reacted for 12 hours. After the reaction was complete, the mixture was cooled to room temperature, water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound T23-1 (449 mg).

[0599] MS m / z(ESI): 258.1 [M+H] + .

[0600] Step 2: 3-(4-aminophenyl)-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T23)

[0601] Compound T23-1 (200 mg, 0.70 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (157 mg, 0.70 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (52 mg, 0.07 mmol), and cesium carbonate (463 mg, 1.41 mmol) were dissolved in dioxane (5 mL) and water (1 mL). The mixture was heated to 90 °C under nitrogen protection and reacted for 12 hours. After the reaction was complete, the mixture was cooled to room temperature, water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the title compound T23 (145 mg).

[0602] MS m / z(ESI): 269.2 [M+H] + .

[0603] Intermediate Preparation Example 24: 3-(4-aminophenyl)-1-(2,2,2-trifluoroethyl)-1H-pyrazolo[3,4-d]pyrimidine-4-ylamine (T24)

[0604]

[0605] Step 1: 3-Bromo-1-(2,2,2-trifluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T24-1)

[0606] Compound T6-1 (100 mg, 0.46 mmol), 2-iodo-1,1,1-trifluoroethane (392 mg, 1.83 mmol), and potassium carbonate (194 mg, 1.37 mmol) were dissolved in N,N-dimethylformamide (3 mL), heated to 80 °C, and reacted for 12 hours. After the reaction was complete, the mixture was cooled to room temperature, water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound T24-1 (140 mg).

[0607] MS m / z(ESI): 296.1 [M+H] + .

[0608] Step 2: 3-(4-aminophenyl)-1-(2,2,2-trifluoroethyl)-1H-pyrazolo[3,4-d]pyrimidine-4-ylamine (T24)

[0609] Compound T24-1 (140 mg, 0.43 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (95 mg, 0.43 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (32 mg, 0.04 mmol), and cesium carbonate (280 mg, 0.85 mmol) were dissolved in dioxane (5 mL) and water (1 mL). The mixture was heated to 90 °C under nitrogen protection and reacted for 12 hours. After the reaction was complete, the mixture was cooled to room temperature, water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the title compound T24 (89 mg).

[0610] MS m / z(ESI): 309.1 [M+H] + .

[0611] Intermediate Preparation Example 25: 5-(4-aminophenyl)-7-(2-fluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T25)

[0612]

[0613] Step 1: 5-Bromo-4-chloro-7-(2-fluoroethyl)-7H-pyrrolo[2,3-d]pyrimidine (T25-1)

[0614] Compound T7-1 (3.0 g, 12.65 mmol), 2-fluoroethyl methanesulfonate (3.0 g, 18.97 mmol), and potassium carbonate (5.35 g, 37.94 mmol) were dissolved in N,N-dimethylformamide (20 mL) and reacted at 85 °C for 8 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to give the title compound T25-1 (2.5 g).

[0615] MS m / z(ESI): 278.0 [M+H] + .

[0616] Step 2: 5-Bromo-7-(2-fluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T25-2)

[0617] Compound T25-1 (1.78 g, 5.74 mmol) was dissolved in dioxane (10 mL), and ammonia (5 mL) was added. The mixture was reacted in a sealed container at 90 °C for 12 hours. After cooling to room temperature, the mixture was evaporated to dryness, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to give the crude product compound T25-2 (1.5 g).

[0618] MS m / z(ESI): 261.1 [M+H] + .

[0619] Step 3: 5-(4-aminophenyl)-7-(2-fluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T25)

[0620] Compound T25-2 (640 mg, 2.47 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (811 mg, 3.71 mmol), cesium carbonate (1.61 g, 4.94 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (180 mg, 0.025 mmol) were dissolved in dioxane / water (15 mL / 1 mL) and reacted at 90 °C for 12 hours under nitrogen. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give the title compound T25 (500 mg).

[0621] MS m / z(ESI): 272.2 [M+H] + .

[0622] Intermediate Preparation Example 26: 3-(4-amino-2-fluorophenyl)-1-(oxetane-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T26)

[0623]

[0624] Compound T9-3 (666 mg, 2.22 mmol), 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (600 mg, 2.44 mmol), cesium carbonate (1.46 g, 4.44 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (166 mg, 0.222 mmol) were dissolved in dioxane / water (5 mL / 1 mL) and reacted under nitrogen atmosphere at 90 °C with microwave for 1.5 h. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give the title compound T26 (400 mg).

[0625] MS m / z (ESI): 301.1 [M+H] + .

[0626] Intermediate Preparation Example 27: 5-(4-aminophenyl)-7-(oxacyclobut-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T27)

[0627]

[0628] Step 1: 5-Bromo-4-chloro-7-(oxetane-3-yl)-7H-pyrrolo[2,3-d]pyrimidine (T27-1)

[0629] Compound T7-1 (600 mg, 2.53 mmol), 3-oxetanebutanol (574 mg, 7.59 mmol), and triphenylphosphine (1.38 g, 5.16 mmol) were dissolved in 1,4-dioxane (4 mL). Diethyl azodicarbonate (917 mg, 5.16 mmol) was added at room temperature. Nitrogen gas was introduced into the reaction vessel, and the mixture was microwaved at 85 °C for 1 hour. After the reaction was complete, the reaction solution was concentrated, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain the title compound T27-1 (350 mg).

[0630] MS m / z(ESI): 289.1 [M+H] + .

[0631] Step 2: 5-Bromo-7-(oxetane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T27-2)

[0632] Compound T27-1 (250 mg, 0.81 mmol) was dissolved in a mixed solvent of ammonia (2 mL) and 1,4-dioxane (1 mL) and reacted in a microwave-safe environment at 90 °C for 1.5 hours. After the reaction was complete, the solution was concentrated, slurried with ethyl acetate, and filtered to obtain the title compound T27-2 (195 mg).

[0633] MS m / z(ESI): 270.1 [M+H] + .

[0634] Step 3: 5-(4-aminophenyl)-7-(oxetane-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T27)

[0635] Compound T27-2 (160 mg, 0.56 mmol), 4-aminophenylboronic acid pinacol ester (126 mg, 0.56 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (42 mg, 0.06 mmol), and cesium carbonate (372 mg, 1.12 mmol) were dissolved in 1,4-dioxane (10 mL) and water (2 mL). The mixture was heated to 90 °C under nitrogen protection and reacted for 12 hours. After the reaction was complete, the mixture was cooled to room temperature, water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the title compound T27 (102 mg).

[0636] MS m / z(ESI): 282.2 [M+H] + .

[0637] Intermediate Preparation Example 28: 5-(4-amino-2-fluorophenyl)-7-(2,2-difluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T28)

[0638]

[0639] Compound T3-3 (615 mg, 2.22 mmol), 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (600 mg, 2.44 mmol), cesium carbonate (1.46 g, 4.44 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (166 mg, 0.222 mmol) were dissolved in dioxane / water (5 mL / 1 mL) and reacted under nitrogen atmosphere at 90 °C with microwave for 1.5 h. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give the title compound T28 (411 mg).

[0640] MS m / z (ESI): 308.1 [M+H] + .

[0641] Intermediate Preparation Example 29: 5-(4-amino-3-fluorophenyl)-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ylamine (T29)

[0642]

[0643] Compound T5-2 (295 mg, 1.00 mmol), 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (242 mg, 1.00 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (75 mg, 0.1 mmol), and cesium carbonate (658 mg, 2 mmol) were added to dioxane (20 mL) and water (4 mL). The mixture was heated to 90 °C under nitrogen protection and reacted for 14 hours. After the reaction was complete, the mixture was cooled to room temperature, concentrated, and separated by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the title compound T29 (242 mg).

[0644] MS m / z(ESI): 326.1 [M+H] + .

[0645] Intermediate Preparation Example 30: 2-(5-fluoropyridin-2-yl)-6-(2-methoxyethyl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T30)

[0646]

[0647] Step 1: (E)-N-cyclohexyl-1,1-dimethoxypropane-2-imine (T30-1)

[0648] Compound T13-1 (40.4 g, 338.6 mmol) and cyclohexylamine (37.3 g, 372.4 mmol) were dissolved in diethyl ether (300 mL), and stirred at 45 °C for half an hour. Calcium chloride (38.0 g, 338.6 mmol) was then added, and the reaction was continued for 5 hours. Thin-layer chromatography (petroleum ether / ethyl acetate = 5 / 1) showed that the reaction was complete. The residue after filtration and concentration was then subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give the title compound T30-1 (50 g).

[0649] Step 2: 1,1,4-Trimethoxybutane-2-one (T30-2)

[0650] T30-1 (42.4 g, 210.8 mmol) was dissolved in tetrahydrofuran (600 mL), and diisopropylaminolithium (231 mL, 1 M) was added at -70 °C. After stirring continuously for 0.5 hours, 1-bromo-2-methoxyethane (40.0 g, 210.8 mmol) was added. The mixture was stirred continuously for 6 hours. Thin-layer chromatography (petroleum ether / ethyl acetate = 5 / 1) showed that the reaction was complete. The mixture was then quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give the title compound T30-2 (30 g).

[0651] Step 3: (E)-5-fluoro-2-(2-(1,1,4-trimethoxybutane-2-yl)hydrazino)pyridine (T30-3)

[0652] Compound 5-fluoro-2-hydrazinopyridine (2.50 g, 17.7 mmol) and T30-2 (3.51 g, 19.5 mmol) were dissolved in methanol (50 mL) and reacted at 60 °C for 2 hours. Thin-layer chromatography (petroleum ether / ethyl acetate = 5 / 1) confirmed the completion of the reaction. The concentrated residue was then subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to separate compound T30-3 (3.0 g).

[0653] MS m / z(ESI): 272.1 [M+H] + .

[0654] Step 4: (E)-2-(2-(5-fluoropyridin-2-yl)hydrazone)-4-methoxybutyraldehyde (T30-4)

[0655] Compound T30-3 (1.0 g, 3.3 mmol) was dissolved in dilute hydrochloric acid (10 mL, 2 N) and reacted at 26 °C for 2 hours. Mass spectrometry showed that the reaction was quenched directly with water after completion, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give the title compound T30-4 (700 mg).

[0656] MS m / z(ESI): 226.3 [M+H]+ .

[0657] Step 5: (E)-5-(2-(2-(5-fluoropyridin-2-yl)hydrazone)-4-methoxybutene)-2,2-dimethyl-1,3-dioxane-4,6-dione (T30-5)

[0658] Compound T30-4 (3.0 g, 12.7 mmol) and 2,2-dimethyl-1,3-dioxane-4,6-dione (2.2 g, 15.2 mmol) were dissolved in toluene (8 mL), and then a mixed solution of tetrahydropyrrole (0.2 mL) and acetic acid (0.2 mL) was added. After stirring at room temperature for 16 hours, the reaction was completed as shown by mass spectrometry. The mixture was then directly filtered, and the solid was the title compound T30-5 (4 g).

[0659] MS m / z (ESI): 352.1 [M+H] + .

[0660] Step Six: 2-(5-Fluoropyridin-2-yl)-6-(2-methoxyethyl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T30)

[0661] Compound T30-5 (1.0 g, 2.7 mmol) was dissolved in methanol (10 mL), and then sodium methoxide (730 mg, 4.0 mmol) was added. After stirring at 65 °C for 2 hours, the reaction was completed according to mass spectrometry. The solvent was removed by concentration, and the pH was adjusted to 6 with dilute hydrochloric acid (1 N). The mixture was then extracted with dichloromethane, and the organic phase was concentrated to obtain the solid, which was the title compound T30 (600 mg).

[0662] MS m / z(ESI): 294 [M+H] + .

[0663] Intermediate Preparation Example 31: 2-(4-chlorophenyl)-6-cyclopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T31)

[0664]

[0665] Step 1: 2-Bromo-1-cyclopropyl-1-ethyl ketone (T31-2)

[0666] Compound T31-1 (3.1 g, 35.0 mmol) was dissolved in methanol (30 mL), and then liquid bromine (5.6 g, 35.0 mmol) was added dropwise under ice bath conditions. The reaction was carried out at 25 °C for 2 hours. Thin-layer chromatography (petroleum ether / ethyl acetate = 5 / 1) showed that after the reaction was complete, water was added, and the mixture was extracted with ethyl acetate, washed with brine, dried over anhydrous sodium sulfate, and concentrated to give the crude title compound T31-2 (6.6 g).

[0667] Step 2: Dimethyl 2-(2-cyclopropyl-2-oxoethyl)malonate (T31-3)

[0668] Dimethyl malonate (5.2 g, 39.0 mmol) was dissolved in tetrahydrofuran (60 mL), and sodium hydride (1.4 g, 35.7 mmol) was added under ice-water bath conditions. After reacting at 0 °C for 0.5 hours, compound T31-2 (5.6 g, 32.5 mmol) was added. Thin-layer chromatography (petroleum ether / ethyl acetate = 5 / 1) showed that the reaction was complete. The mixture was quenched with water, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. The concentrated residue was then subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give the title compound T31-3 (4 g).

[0669] Step 3: Methyl 2-(4-chlorophenyl)-6-cyclopropyl-3-oxo-2,3,4,5-tetrahydropyridazine-4-carboxylate (T31-4)

[0670] Compound T31-3 (1.1 g, 5.0 mmol) and 4-chlorophenylhydrazine (0.8 g, 5.5 mmol) were dissolved in acetic acid (20 mL), and sodium acetate (2.0 g, 15.0 mmol) was added. The reaction was carried out at 50 °C for 16 hours. Thin-layer chromatography (petroleum ether / ethyl acetate = 5 / 1) confirmed the completion of the reaction. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was adjusted to a weakly alkaline pH with saturated sodium bicarbonate. The organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated to give crude compound T31-4 (1.6 g).

[0671] MS m / z(ESI): 307 [M+H] + .

[0672] Step 4: Methyl 2-(4-chlorophenyl)-6-cyclopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylate (T31-5)

[0673] Compound T31-4 (1.4 g, 4.3 mmol) was dissolved in acetonitrile (20 mL), and copper chloride (1.58 g, 8.7 mmol) was added. The reaction was carried out at 75 °C for 2 hours. After the reaction was completed, the solvent was removed by rotary evaporation, and the title compound T31-5 (496 mg) was purified by silica gel column chromatography.

[0674] MS m / z(ESI): 305 [M+H] + .

[0675] Step 5: 2-(4-Chlorophenyl)-6-cyclopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T31)

[0676] Compound T31-5 (496 mg, 1.5 mmol) and lithium hydroxide (196 mg, 4.6 mmol) were dissolved in a mixed solution of methanol (8 mL) and water (2 mL). After stirring at room temperature for 1 hour, the mass spectrometry showed that the reaction was complete. The solution was then concentrated to remove methanol. The crude product was then mixed with water and the pH was adjusted to weakly acidic with dilute hydrochloric acid. A solid precipitated out and was filtered. The solid was the title compound T31 (0.4 g).

[0677] MS m / z(ESI): 291 [M+H] + .

[0678] Intermediate Preparation Example 32: 2-(5-chloropyridin-2-yl)-6-cyclopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T32)

[0679]

[0680] Step 1: (E)-2-(2-(2-(5-chloropyridin-2-yl)hydrazone)-2-cyclopropylethyl)dimethyl malonate (T32-1)

[0681] Compound T31-2 (2.4 g, 11.1 mmol) was dissolved in methanol (20 mL) with 5-chloro-2-hydrazinopyridine (1.7 g, 11.6 mmol) and one drop of acetic acid, and reacted at 80 °C for 16 hours. After the reaction was completed, the solvent was removed by rotary evaporation to obtain the crude title compound T32-1 (3.8 g).

[0682] MS m / z(ESI): 340 [M+H] + .

[0683] Step 2: Methyl 2-(5-chloropyridin-2-yl)-6-cyclopropyl-3-oxo-2,3,4,5-tetrahydropyridazine-4-carboxylate (T32-2)

[0684] The crude compound T32-1 (3.8 g, 11.1 mmol) was dissolved in acetic acid (20 mL), followed by the addition of sodium acetate (4.5 g, 33.2 mmol). The reaction was carried out at 50 °C for 16 hours. Mass spectrometry showed that after the reaction was complete, the mixture was extracted with ethyl acetate and saturated sodium bicarbonate was added to adjust the pH to weakly alkaline. The mixture was then washed with brine, dried over anhydrous sodium sulfate, and concentrated to obtain the crude title compound T32-2 (3.5 g).

[0685] MS m / z(ESI): 308 [M+H] + .

[0686] Step 3: Methyl 2-(5-chloropyridin-2-yl)-6-cyclopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylate (T32-3)

[0687] The crude compound T32-2 (3.5 g, 10.8 mmol) was dissolved in acetonitrile (20 mL), and then copper chloride (2.2 g, 16.2 mmol) was added. The reaction was carried out at 80 °C for 4 hours. After the reaction was completed, the solvent was removed by rotary evaporation, and the title compound T32-3 (2.2 g) was purified by silica gel column chromatography.

[0688] MS m / z(ESI): 306 [M+H] + .

[0689] Step 4: 2-(5-chloropyridin-2-yl)-6-cyclopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T32)

[0690] Compound T32-3 (2.2 g, 6.7 mmol) was dissolved in a mixture of methanol (20 mL) and water (5 mL), and then lithium hydroxide (494 mg, 20.2 mmol) was added. The reaction was carried out at 25 °C for 0.5 h. After the reaction was completed, the solution was directly concentrated to remove methanol, and the pH was adjusted to weakly acidic with dilute hydrochloric acid. The precipitated solid was the title compound T32 (0.1 g).

[0691] MS m / z(ESI): 292 [M+H] + .

[0692] Preparation example:

[0693] Example 1: N-(4-(4-amino-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 1)

[0694]

[0695] Step 1: 3-Bromo-1-(4-methoxybenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (1-1)

[0696] Compound T6-1 (5.0 g, 23.41 mmol), p-methoxybenzyl chloride (3.4 g, 21.07 mmol), and potassium carbonate (6.6 g, 46.82 mmol) were added to N,N-dimethylformamide (50 mL), and the reaction was carried out at 25 °C for 16 hours. The reaction was quenched with water, and a solid was obtained. The solid was directly filtered to give the title compound 1-1 (5.12 g).

[0697] MS m / z (ESI): 336.1 [M+H] + .

[0698] Step 2: 3-(4-aminophenyl)-1-(4-methoxybenzyl)-1H-pyrazolo[3,4-d]pyrimidine-4-ylamine (1-2)

[0699] Compound 1-1 (2.5 g, 6.34 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (1.56 g, 6.98 mmol), sodium carbonate (1.37 g, 12.68 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (530 mg, 0.634 mmol) were added to dioxane / water (20 mL / 4 mL) and reacted at 110 °C for 5 hours under nitrogen. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give the title compound 1-2 (1.3 g).

[0700] MS m / z (ESI): 347.2 [M+H] + .

[0701] Step 3: N-(4-(4-amino-1-(4-methoxybenzyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (1-3)

[0702] Compounds 1-2 (1.0 g, 2.71 mmol), 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (824 mg, 2.71 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.58 g, 4.07 mmol), and N,N-diisopropylethylamine (1.07 g, 8.14 mmol) were added to N,N-dimethylformamide (20 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and a solid precipitated. The filter cake was washed with water, dissolved in ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give the title compound 1-3 (1.48 g).

[0703] MS m / z (ESI): 602.2 [M+H] + .

[0704] Step 4: N-(4-(4-amino-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (1-4)

[0705] Compounds 1-3 (1.48 g, 2.19 mmol) were added to trifluoroacetic acid (10 mL) and reacted at 70 °C for 8 hours. The reaction was quenched with saturated sodium carbonate solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and concentrated to give the title compound 1-4 (1.1 g).

[0706] MS m / z(ESI): 482.2 [M+H] + .

[0707] Step 5: N-(4-(4-amino-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 1)

[0708] Compounds 1-4 (116 mg, 0.207 mmol), methyl (2-fluorophenyl)methanesulfonate (106 mg, 0.311 mmol), and potassium carbonate (87 mg, 0.623 mmol) were added to N,N-dimethylformamide (5 mL), and the mixture was reacted at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain compound 1 (25 mg).

[0709] MS m / z(ESI): 590.2 [M+H] + .

[0710] 1 H NMR (400MHz, DMSO-d6) δ11.74(s,1H),8.50(s,1H),8.33(d,J=10.8Hz,2H),7.91(t,J=6.8Hz,3H),7.70(d,J=8.4Hz,2H),7.61( d,J=8.0Hz,1H),7.38(dd,J=14.0,6.8Hz,1H),7.30-7.13(m,3H),5.64(s,2H),3.12(m,1H),2.44(s,3H),1.27(d,J=6.8Hz,6H).

[0711] Example 2: N-(4-(4-amino-1-((4-oxocyclohexyl)methyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 2)

[0712]

[0713] Step 1: 4-((4-amino-3-bromo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)cyclohexanone (2-1)

[0714] Compound T6-1 (510 mg, 2.34 mmol), 4-(hydroxymethyl)cyclohexanone (460 mg, 3.50 mmol), and triphenylphosphine (1.25 g, 4.67 mmol) were added to tetrahydrofuran (10 mL). Diethyl azodicarbonate (830 mg, 4.67 mmol) was added dropwise under nitrogen atmosphere in an ice-water bath, and the reaction was carried out at 25 °C for 5 hours. The reaction was quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give the crude compound 2-1 (700 mg).

[0715] MS m / z(ESI): 324.1 [M+H] + .

[0716] Step 2: 4-((4-amino-3-(4-aminophenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)cyclohexanone (2-2)

[0717] Compound 2-1 (700 mg, 2.31 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (620 mg, 2.78 mmol), sodium carbonate (500 mg, 4.63 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (193 mg, 0.231 mmol) were added to dioxane / water (10 mL / 2 mL) and reacted at 110 °C for 5 hours under nitrogen. After cooling to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give the title compound 2-2 (500 mg).

[0718] MS m / z (ESI): 337.3 [M+H] + .

[0719] Step 3: N-(4-(4-amino-1-((4-oxocyclohexyl)methyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 2)

[0720] Compound 2-2 (500 mg, 1.49 mmol), 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (450 mg, 1.49 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (865 mg, 2.23 mmol), and diisopropylethylamine (588 mg, 4.46 mmol) were added to N,N-dimethylformamide (10 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 2 (50 mg).

[0721] MS m / z (ESI): 592.3 [M+H] + .

[0722] 1 H NMR (400MHz, DMSO-d6) δ11.75(s,1H),8.51(s,1H),8.35(s,1H),8.28(s,1H),7.92(d,J=8.4Hz,3H),7.73(d,J=8.4Hz,2H),7.61(d,J=8.0Hz,1H),4. 35(d,J=7.2Hz,2H),3.13(m,1H),2.44(s,3H),2.37(m,2H),2.23(d,J=14. 4Hz,2H),1.88(d,J=10.0Hz,2H),1.57-1.45(m,2H),1.28(d,J=6.8Hz,6H).

[0723] Example 3: N-(4-(4-amino-7-(oxacyclobut-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 3)

[0724]

[0725] Compound T9 (100 mg, 337 μmol) and 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (97 mg, 337 μmol) were weighed and dissolved in N,N-dimethylformamide (2 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (154 mg, 404 μmol) and diisopropylethylamine (87 mg, 673 μmol) were added, and the mixture was stirred at 25 °C for 5 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 3 (23 mg).

[0726] MS m / z (ESI): 538.2 [M+H] + .

[0727] 1 H NMR (400MHz, DMSO-d6) δ11.77(s,1H),8.51(s,1H),8.35(s,1H),8.28(s,1H),7.95-7.92(m,3H),7.78(d,J=8.0Hz,2H),7.61(d,J= 8.0Hz,1H),6.09-6.02(m,1H),5.12(t,J=6.4Hz,2H),5.02(t,J=6.4Hz,2H),3.17-3.10(m,1H),2.42(s,3H),1.28(d,J=8.0Hz,6H).

[0728] Example 4: N-(4-(4-amino-1-cyclopropylmethyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 4)

[0729]

[0730] Compounds 1-4 (33.30 mg, 62.30 μmol), cyclopropylmethyl bromide (10.30 mg, 74.77 μmol), and potassium carbonate (26.40 mg, 186.91 μmol) were added to dimethyl sulfone (2 mL) and reacted at 70 °C for 2 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 4 (12 mg).

[0731] MS m / z (ESI): 536.3 [M+H] + .

[0732] 1H NMR (400MHz, DMSO-d6) δ8.48(s,1H),8.32(s,1H),8.24(s,1H),7.89(d,J=8.0Hz,3H),7.69(d,J=8.0Hz,2H),7.58(d,J=8.0Hz,1H),4 .21(d,J=8.0Hz,2H),3.14-3.07(m,1H),2.41(s,3H),1.31-1.27(m,1H),1.25(d,J=8.0Hz,6H),0.50-0.47(m,2H),0.44-0.42(m,2H).

[0733] Example 5: N-(4-(4-amino-1-(4-cyanocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 5)

[0734]

[0735] Step 1: 4-(4-amino-3-bromo-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanecarboxylonite (5-1)

[0736] Compound T6-1 (250 mg, 1.17 mmol) was weighed and dissolved in tetrahydrofuran (10 mL). 4-Hydroxycyclohexanenitrile (219 mg, 1.75 mmol) and triphenylphosphine (613 mg, 2.34 mmol) were added, followed by diethyl azodicarbonate (409 mg, 2.34 mmol) at 0 °C. The mixture was stirred at 25 °C for 16 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography to obtain the title compound 5-1 (220 mg).

[0737] MS m / z(ESI): 321.2 [M+H] + .

[0738] Step 2: 4-[4-amino-3-(4-aminophenyl)pyrazolo[3,4-d]pyrimidin-1-yl]cyclohexanecarboxylonite (compound 5-2)

[0739] Compound 5-1 (220 mg, 582 μmol) was weighed and dissolved in 1,4-dioxane (5 mL) and water (1 mL). 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)aniline (153 mg, 699 μmol), sodium carbonate (123 mg, 1.16 mmol), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane dichloride complex (95 mg, 116 μmol) were added. The mixture was stirred at 90 °C for 16 hours under nitrogen protection. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography to obtain the title compound 5-2 (180 mg).

[0740] MS m / z(ESI): 334.2 [M+H] + .

[0741] Step 3: N-(4-(4-amino-1-(4-cyanocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 5)

[0742] Compound 5-1 (80 mg, 228 μmol) and 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (97 mg, 337 μmol) were weighed and dissolved in N,N-dimethylformamide (2 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (104 mg, 274 μmol) and diisopropylethylamine (89 mg, 673 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 5 (65 mg).

[0743] MS m / z (ESI): 589.2 [M+H] + .

[0744] 1H NMR (400MHz, DMSO-d6) δ11.75(s,1H),8.50(s,1H),8.35(s,1H),8.27(s,1H),7.93-7.91(m,3H),7.72(d,J=8.0Hz,2H),7.60(d,J=8.0Hz,1H),4.8 3-4.78(m,1H),3.30-3.25(m,1H),3.18-3.10(m,1H),2.45(s,3H),2.25- 2.15(m,2H),2.10-1.95(m,4H),1.95-1.90(m,2H),1.28(d,J=8.0Hz,6H).

[0745] Example 6: N-(4-(4-amino-1-(oxacyclobut-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-methylpyridin-2-yl)-3-oxo-6-(((tetrahydro-2H-pyran-4-yl)methyl)-2,3-dihydropyridazine-4-carboxamide (Compound 6)

[0746]

[0747] Weigh out T9 (50 mg, 159 μmol) and 2-(5-methylpyridin-2-yl)-3-oxo-6-((tetrahydro-2H-pyran-4-yl)methyl)-2,3-dihydropyridazine-4-carboxylic acid (T12) (55 mg, 159 μmol), dissolve them in N,N-dimethylformamide (2 mL), add triethylamine (41 mg, 319 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (73 mg, 191 μmol), and stir at 25 °C for 2 hours. Add water to the reaction solution, extract with ethyl acetate, and combine the organic phases. Wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and separate the crude product by silica gel column chromatography to obtain the title compound 6 (22 mg).

[0748] MS m / z (ESI): 594.2 [M+H] + .

[0749] 1H NMR(400MHz,DMSO-d6)δ11.79(s,1H),8.50(s,1H),8.3(s,1H),8.27(s,1H),8.28(s, 1H),7.94-7.91(m,3H),7.77-7.65(m,2H),7.61(d,J=8.0Hz,1H),6.05-6.03(m,1H),5 .12(t,J=6.0Hz,2H),5.01(t,J=6.0Hz,2H),3.87-3.83(m,2H),3.34-3.28(m,2H),2.7 2-2.70(m,2H),2.44(s,3H),2.10-1.90(m,1H),1.62-1.58(m,2H),1.32-1.25(m,2H).

[0750] Example 7: N-(4-(4-amino-1-isobutyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 7)

[0751]

[0752] Compound T20 (45 mg, 0.141 mmol), 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (40 mg, 0.141 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (82 mg, 0.212 mmol), and diisopropylethylamine (56 mg, 0.425 mmol) were added to N,N-dimethylformamide (5 mL), and the mixture was reacted at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 7 (25 mg).

[0753] MS m / z (ESI): 538.3 [M+H] + .

[0754] 1H NMR (400MHz, DMSO-d6) δ11.73(s,1H),8.50(s,1H),8.34(s,1H),8.27(s,1H),7.94-7.88(m,3H),7.71(d,J=8.4Hz,2H),7.61(d,J=8.0Hz ,1H),4.18(d,J=7.2Hz,2H),3.17-3.08(m,1H),2.44(s,3H),2.28(dq,J=13.6,6.8Hz,1H),1.28(d,J=6.8Hz,6H),0.90(d,J=6.8Hz,6H).

[0755] Example 8: N-(4-(4-amino-1-(oxacyclobut-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-3-fluorophenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 8)

[0756]

[0757] Compound T26 (75 mg, 0.233 mmol), 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (62 mg, 0.221 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (136 mg, 0.349 mmol), and N,N-diisopropylethylamine (92 mg, 0.699 mmol) were added to N,N-dimethylformamide (5 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 8 (25 mg).

[0758] MS m / z (ESI): 556.1 [M+H] + .

[0759] 1 H NMR(400MHz,DMSO-d6)δ11.84(s,1H),8.51(s,1H),8.35(s,1H),8.26(s,1H),7.99(m,1H),7.93(m,1H),7.69-7.56(m,3H ),6.12-5.96(m,1H),5.10(t,J=6.4Hz,2H),5.02(t,J=7.2Hz,2H),3.18-3.09(m,1H),2.45(s,3H),1.28(d,J=7.2Hz,6H).

[0760] Example 9: N-(4-(4-amino-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 9)

[0761]

[0762] Compound T21 (74 mg, 0.238 mmol), 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (92 mg, 0.286 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (139 mg, 0.358 mmol), and N,N-diisopropylethylamine (94 mg, 0.716 mmol) were added to N,N-dimethylformamide (5 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 9 (5 mg).

[0763] MS m / z (ESI): 546.1 [M+H] + .

[0764] 1 H NMR (400MHz, DMSO-d6) δ11.75(s,1H),8.51(s,1H),8.33(d,J=10.8Hz,2H),7.93(d,J=8.4Hz,3H),7.73(d,J=8.4Hz,2H),7.61 (d,J=8.0Hz,1H),6.51(s,1H),4.82(dt,J=14.8,7.6Hz,2H),3.13(dt,J=13.6,6.8Hz,1H),2.44(s,3H),1.28(d,J=6.8Hz,6H).

[0765] Example 10: N-(4-(4-amino-1-(oxacyclobut-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-2-fluorophenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 10)

[0766]

[0767] Compound T11 (120 mg, 0.32 mmol), 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (92 mg, 0.32 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 0.38 mmol), and diisopropylethylamine (84 mg, 0.64 mmol) were added to N,N-dimethylformamide (3 mL), and the mixture was stirred at 25 °C for 5 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with water and saturated brine, respectively, and dried over anhydrous sodium sulfate. The concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 10 (15 mg).

[0768] MS m / z (ESI): 556.1 [M+H] + .

[0769] 1 H NMR (400MHz, DMSO-d6) δ12.17(s,1H),8.74(t,J=8.4Hz,1H),8.51(s,1H),8.42(s,1H),8.28(s,1H),7.92(dd,J=8.0,2.0Hz,1H),7.68 -7.61(m,3H),6.08-6.01(m,1H),5.12(t,J=6.8Hz,2H),5.01(t,J=7.6Hz,2H),3.20-3.10(m,1H),2.44(s,3H),1.05(d,J=7.2Hz,6H).

[0770] Example 11: N-(4-(4-amino-1-(2-(methanesulfonyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 11)

[0771]

[0772] Step 1: 3-Bromo-1-(2-(Methylsulfonyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (11-1)

[0773] Compound T6-1 (204.08 mg, 934.48 μmol), ethyl 2-(methanesulfonyl)methanesulfonate, and potassium carbonate (210.00 mg, 934.48 μmol) were added to N,N-dimethylformamide (10 mL), and the mixture was heated to 70 °C and reacted for 12 hours. After the reaction was complete, the mixture was cooled to room temperature, water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, concentrated, and then slurried with dichloromethane to obtain the title compound 11-1 (210 mg).

[0774] MS m / z (ESI): 320.1 [M+H] + .

[0775] Step 2: 3-(4-aminophenyl)-1-(2-(methanesulfonyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (11-2)

[0776] Compound 11-1 (210 mg, 590.32 μmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (131.97 mg, 590.32 μmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (44.08 mg, 59.03 μmol), and cesium carbonate (388.56 mg, 1.18 mmol) were added to dioxane (15 mL) and water (3 mL). The mixture was heated to 90 °C under nitrogen protection and reacted for 12 hours. After the reaction was complete, the mixture was cooled to room temperature, concentrated, and separated by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to obtain the title compound 11-2 (325 mg).

[0777] MS m / z(ESI): 333.1 [M+H] + .

[0778] Step 3: N-(4-(4-amino-1-(2-(methanesulfonyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 11)

[0779] Compound 11-2 (63 mg, 0.19 mmol), 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (55 mg, 0.19 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (90 mg, 0.23 mmol), and diisopropylethylamine (51 mg, 0.39 mmol) were added to N,N-dimethylformamide (3 mL), and the mixture was stirred at 25 °C for 5 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with water and saturated brine, respectively, and dried over anhydrous sodium sulfate. The concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 11 (29 mg).

[0780] MS m / z (ESI): 588.2 [M+H] + .

[0781] 1 H NMR (400MHz, DMSO-d6) δ11.74(s,1H),8.50(s,1H),8.32(d,J=12.8Hz,2H),7.92(m,3H),7.72(m,2H),7.61(d ,J=7.6Hz,1H),4.80(m,2H),3.80(m,2H),3.22-3.09(m,1H),3.04(s,3H),2.44(s,3H),1.28(d,J=6.4Hz,6H).

[0782] Example 12: N-(4-(4-amino-1-(trans-4-hydroxycyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazin-4-carboxamide (compound 12A) and N-(4-(4-amino-1-(cis-4-hydroxycyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazin-4-carboxamide (compound 12B)

[0783]

[0784] Step 1: 1,4-Dioxaspiro[4.5]dec-8-ylmethanesulfonate (12-2)

[0785] Compound 12-1 (3 g, 18 mmol) was weighed and added to dichloromethane (60 mL). Triethylamine (3.65 g, 36 mmol) and methanesulfonic anhydride (4.71 g, 27 mmol) were added at 0 °C, and the mixture was stirred at 0 °C for 1 hour. Water was added to the reaction solution, and the mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound 12-2 (4 g).

[0786] Step 2: 3-Bromo-1-(1,4-dioxaspiro[4.5]dec-8-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (12-3)

[0787] 2 g (9.34 mmol) of 3-bromo-1H-pyrazolo[3,4-d]pyrimidine-4-amine was weighed and added to 40 mL of N,N-dimethylformamide, followed by 9.13 g (28 mmol) of cesium carbonate and 3.49 g (14 mmol) of compound 12-2. The mixture was stirred at 80 °C for 16 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and slurried with dichloromethane to give compound 12-3 (3 g).

[0788] MS m / z (ESI): 354.1 [M+H] + .

[0789] Step 3: 3-(4-aminophenyl)-1-(1,4-dioxaspiro[4.5]dec-8-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (12-4)

[0790] Compound 12-3 (2 g, 4.52 mmol) was weighed and added to water (5 mL) and 1,4-dioxane (25 mL), followed by [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (661 mg, 903 μmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)aniline (1.29 g, 5.87 mmol), and sodium carbonate (9581 mg, 9.03 mmol). The mixture was stirred at 90 °C for 16 hours under nitrogen protection. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography to obtain the title compound 12-4 (1.6 g).

[0791] MS m / z (ESI): 367.1 [M+H] + .

[0792] Step 4: N-(4-(4-amino-1-(1,4-dioxaspiro[4.5]dec-8-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (12-5)

[0793] Compound 12-4 (211 mg, 546 μmol) and 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (497 mg, 546 μmol) were weighed and added to N,N-dimethylformamide (5 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (249 mg, 655 μmol) and triethylamine (141 mg, 1.09 mmol) were added, and the mixture was stirred at 25 °C for 5 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by column chromatography to obtain the title compound 12-5 (300 mg).

[0794] MS m / z(ESI): 622.1 [M+H] + .

[0795] Step 5: N-(4-(4-amino-1-(4-oxocyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (12-6)

[0796] Compound 12-5 (150 mg, 181 μmol) was weighed and added to trifluoroacetic acid (5 mL), water (1 mL), and dichloromethane (5 mL). The mixture was stirred at 25 °C for 5 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by column chromatography to obtain the title compound 12-6 (100 mg).

[0797] MS m / z(ESI): 578.2 [M+H] + .

[0798] Step Six: N-(4-(4-amino-1-(4-hydroxycyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-1,2,3,6-tetrahydropyridazine-4-carboxamide (12-7)

[0799] Compound 12-6 (100 mg, 139 μmol) was weighed and dissolved in methanol (2 mL). NaBH4 (10 mg, 277 μmol) was added at 0 °C, and the mixture was stirred at 0 °C for 1 hour. Water was added to the reaction solution, and the mixture was extracted with dichloromethane, washed with saturated brine, and dried over anhydrous sodium sulfate to give the title compound 12-7 (100 mg).

[0800] MS m / z (ESI): 582.2 [M+H] + .

[0801] Step 7: N-(4-(4-amino-1-(trans-4-hydroxycyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide and N-(4-(4-amino-1-(cis-4-hydroxycyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide)

[0802] Compound 12-7 (80 mg, 110 μmol) was weighed and added to methanol (3 mL), followed by palladium / carbon (10 mg, 10% pure). The mixture was stirred at 50 °C for 16 hours in air. The reaction solution was directly filtered, concentrated, and the crude product was separated by TLC to obtain the title compound.

[0803] The following are the physicochemical parameters of the two isomers mentioned above:

[0804] Compound 12-I (dichloromethane / methanol = 10 / 1, Rf: 0.54, 6.48 mg):

[0805] MS m / z (ESI): 580.2 [M+H] + .

[0806] 1 H NMR (400MHz, DMSO-d6) δ11.74(s,1H),8.51(s,1H),8.35(s,1H),8.26(s,1H),7.93-7.90(m,3H),7.74-7.70(m,2H),7.61(d,J=8.0Hz ,1H),4.73-4.65(m,2H),3.60-3.50(m,1H),3.20-3.10(m,1H),2.44(s,3H),2.15-1.95(m,4H),1.50-1.40(m,2H),1.28-1.26(m,6H).

[0807] Compound 12-II (dichloromethane / methanol = 10 / 1, Rf: 0.43, 3.27 mg):

[0808] MS m / z (ESI): 580.2 [M+H] + .

[0809] 1 H NMR(400MHz,DMSO-d6)δ11.73(s,1H),8.50(s,1H),8.34(s,1H),8.25(s,1H ),7.93-7.90(m,3H),7.74-7.70(m,2H),7.61(d,J=8.0Hz,1H),4.78-4.69(m ,1H),3.95-3.90(m,1H),3.20-3.10(m,1H),2.44(s,3H),2.43-2.34(m,2H), 2.10-2.00(m,1H),1.90-1.80(m,2H),1.70-1.60(m,4H),1.28-1.26(m,6H).

[0810] Example 13: N-(4-(4-amino-1-(2-fluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 14)

[0811]

[0812] Compound T22 (200 mg, 0.580 mmol), 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (130 mg, 0.464 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (337 mg, 0.869 mmol), and diisopropylethylamine (230 mg, 1.74 mmol) were added to N,N-dimethylformamide (10 mL) and reacted at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate. The concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 14 (70 mg).

[0813] MS m / z (ESI): 528.2 [M+H] + .

[0814] 1H NMR(400MHz,DMSO-d6)δ11.74(s,1H),8.51(d,J=1.6Hz,1H),8.34(s,1H),8 .29(s,1H),7.92(d,J=8.4Hz,3H),7.73(d,J=8.4Hz,2H),7.61(d,J=8.0Hz, 1H),4.97(t,J=4.8Hz,1H),4.85(t,J=4.8Hz,1H),4.71(t,J=4.8Hz,1H),4. 65(t,J=4.8Hz,1H),3.17-3.08(m,1H),2.44(s,3H),1.28(d,J=6.8Hz,6H).

[0815] Example 14: N-(4-(4-amino-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 15)

[0816]

[0817] Compound T10 (100 mg, 321 μmol) and 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (88 mg, 321 μmol) were weighed and added to N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were then added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 15 (75 mg).

[0818] MS m / z (ESI): 552.4 [M+H] + .

[0819] 1H NMR (400MHz, DMSO-d6) δ11.74(s,1H),8.50(s,1H),8.35(s,1H),8.28-8.22(m,1H),7.91(m,3H),7.71(d,J=8.4Hz,2H),7.61(d,J= 8.0Hz,1H),5.56-5.50(m,1H),4.18-4.04(m,2H),3.99-3.89(m,2H),3.18-3.08(m,1H),2.48-2.38(m,5H),1.28(d,J=6.8Hz,6H).

[0820] Example 15: N-(4-(4-amino-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 16)

[0821]

[0822] Step 1: 2-(4-amino-3-bromo-pyrazolo[3,4-d]pyrimidin-1-yl)ethanol (16-1)

[0823] Compound T6-1 (3.03 g, 14 mmol) and 2-iodoethanol (2.68 g, 15.4 mmol) were added to N,N-dimethylformamide (10 mL), followed by potassium carbonate (3.91 g, 28 mmol). The mixture was stirred at 80 °C for 2 hours. Water was added to the reaction mixture, followed by extraction with ethyl acetate, washing with saturated brine, drying with anhydrous sodium sulfate, and concentrating. The crude product was subjected to column chromatography to obtain the title compound 16-2 (1 g).

[0824] MS m / z(ESI): 257.1 [M+H] + .

[0825] Step 2: 2-[4-amino-3-(4-aminophenyl)pyrazolo[3,4-d]pyrimidin-1-yl]ethanol (16-2)

[0826] Compound 16-1 (500 mg, 1.74 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)aniline (463 mg, 2.09 mmol) were dissolved in N,N-dimethylformamide (10 mL). Sodium carbonate (373 mg, 3.49 mmol) and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (129 mg, 174 μmol) were added, and the mixture was reacted at 110 °C for 5 hours under nitrogen protection. The mixture was filtered with diatomaceous earth, the filtrate was washed with ethyl acetate, the resulting organic phase was dried over anhydrous sodium sulfate, concentrated, and the crude product was subjected to column chromatography to obtain the title compound 16-3 (200 mg).

[0827] MS m / z(ESI): 271.1 [M+H] + .

[0828] Step 3: N-(4-(4-amino-1-(2-hydroxyethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 16)

[0829] Compound 16-2 (64 mg, 0.21 mmol), 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (59 mg, 0.19 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (82 mg, 0.21 mmol), and diisopropylethylamine (50 mg, 0.39 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 16 (10 mg).

[0830] MS m / z (ESI): 526.3 [M+H] + .

[0831] 1H NMR (400MHz, DMSO-d6) δ11.73(s,1H),8.51(s,1H),8.35(s,1H),8.27(s,1H),7.91(d,J=8.4Hz,3H),7.72(d,J=8.4Hz,2H),7.61(d,J =8.0Hz,1H),4.92(s,1H),4.40(q,J=7.2Hz,2H),3.87-3.88(m,2H),3.12(dd,J=13.6,6.8Hz,1H),2.45(s,3H),1.28(d,J=6.8Hz,6H).

[0832] Example 16: N-(4-(4-amino-1-(oxacyclobut-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 17)

[0833]

[0834] Compound T9 (100 mg, 319 μmol) and 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14) (98 mg, 319 μmol) were weighed and added to N,N-dimethylformamide (4 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (148 mg, 383 μmol) and diisopropylethylamine (84 mg, 638 μmol) were added, and the mixture was stirred at 25 °C for 5 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 17 (36 mg).

[0835] MS m / z (ESI): 541.1 [M+H] + .

[0836] 1 H NMR(400MHz,DMSO-d6)δ11.82(s,1H),8.32(s,1H),8.28(s,1H),7.96-7.94(m,2H),7.78-7.76(m,2H),7.75-7.71(m,2H),7 .45-7.41(m,2H),6.06-6.03(m,1H),5.12(t,J=6.4Hz,2H),5.02(t,J=6.4Hz,2H),3.17-3.10(m,1H),1.28(d,J=8.0Hz,6H).

[0837] Example 17: N-(4-(4-amino-1-isobutyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 18)

[0838]

[0839] Compound T20 (100 mg, 336 μmol) and 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14) (103 mg, 336 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (148 mg, 404 μmol) and diisopropylethylamine (89 mg, 673 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 18 (71 mg).

[0840] MS m / z (ESI): 541.1 [M+H] + .

[0841] 1 H NMR(400MHz,DMSO-d6)δ11.80(s,1H),8.31(s,1H),8.27(s,1H),7.93-7.91(m,2H),7.74-7.71 (m,4H),7.45-7.40(m,2H),4.18(d,J=7.2Hz,2H),3.15-3.10(m,1H),2.33-2.26(m,1H),1.29(d,J=6.8Hz,6H),0.91(d,J=6.8Hz,6H).

[0842] Example 18: N-(4-(4-amino-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 19)

[0843]

[0844] Compound T10 (100 mg, 321 μmol) and 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14) (98 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 19 (64 mg).

[0845] MS m / z (ESI): 555.2 [M+H] + .

[0846] 1 H NMR(400MHz,DMSO-d6)δ11.80(s,1H),8.32(s,1H),8.28(s,1H),7.93-7.92(m,2H),7.74-7.71(m,4H),7.45-7.40(m,2H), 5.56-5.50(m,1H),4.16-4.08(m,2H),3.99-3.89(m,2H),3.17-3.10(m,1H),2.46(q,J=6.8Hz,2H),1.29(d,J=7.2Hz,6H).

[0847] Example 19: N-[4-(4-amino-1-tetrahydrofuran-3-yl-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl]-6-isopropyl-3-oxo-2-[5-(trifluoromethyl)pyridin-2-yl]-2,3-dihydropyridazine-4-carboxamide (Compound 20)

[0848]

[0849] Weigh out T10 (100 mg, 321 μmol) and 6-isopropyl-3-oxo-2-[5-(trifluoromethyl)-2-pyridyl]-2,3-dihydropyridazine-4-carboxylic acid (T16) (117 mg, 321 μmol), dissolve in N,N-dimethylformamide (4 mL), add diisopropylethylamine (127 mg, 962 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 385 μmol), and stir at 25 °C for 2 hours. Add water to the reaction solution, extract with ethyl acetate, and combine the organic phases. Wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and separate the crude product by preparative high performance liquid chromatography to obtain the title compound 20 (42 mg).

[0850] MS m / z (ESI): 606.2 [M+H] + .

[0851] 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),9.15(s,1H),8.57(dd,J=2.0,8.0Hz,1H),8.35(s,1H),8.28(s,1H),8.04(d,J=8.0Hz,1H),7.92(d,J=8.0Hz ,2H),7.72(d,J=8.0Hz,2H),5.54-5.54(m,1H),4.16-4.08(m,2H),3.99- 3.89(m,2H),3.18-3.13(m,1H),2.46-2.41(m,2H),1.29(d,J=7.2Hz,6H).

[0852] Example 20: N-(4-(4-amino-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-3-oxo-2-(4-(trifluoromethyl)phenyl)-2,3-dihydropyridazine-4-carboxamide (Compound 21)

[0853]

[0854] Compound T19 (30 mg, 82.5 μmol), 3-(4-aminophenyl)-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine (T10) (26 mg, 82.5 μmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (48 mg, 123 μmol), and N,N'-diisopropylethylamine (33 mg, 246 μmol) were added to N,N'-dimethylformamide (5 mL), and the mixture was stirred at 25 °C for 16 hours. Water was added, and the mixture was extracted with ethyl acetate and dried over anhydrous sodium sulfate. The concentrated residue was separated by preparative high performance liquid chromatography to obtain the title compound 21 (6 mg).

[0855] MS m / z (ESI): 605.2 [M+H] + .

[0856] 1 H NMR (400MHz, DMSO-d6) δ11.66(s,1H),8.30(s,1H),8.26(s,1H),7.99-7.85(m,6H),7.70(d,J=8.4Hz ,2H),5.56-5.45(m,1H),4.10(m,2H),3.93(m,2H),3.12(m,1H),2.41(m,2H),1.28(d,J=6.8Hz,6H).

[0857] Example 21: N-(4-(4-amino-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 22)

[0858]

[0859] Compound T10 (65 mg, 0.208 mmol), 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17) (63 mg, 0.208 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (121 mg, 0.312 mmol), and N,N-diisopropylethylamine (83 mg, 0.625 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 22 (33 mg).

[0860] MS m / z(ESI): 571.1 [M+H]+ .

[0861] 1 H NMR (400MHz, DMSO-d6) δ11.75(s,1H),8.29(d,J=7.2Hz,2H),7.91(d,J=8.0Hz,2H),7.72(d,J=4.8Hz,4H),7.65(d,J=8.4Hz,2H),5.58-5.49 (m,1H),4.18-4.06(m,2H),3.95(ddd,J=21.2,11.6,5.6Hz,2H),3.13(dt,J=13.6,6.8Hz,1H),2.44(q,J=6.8Hz,2H),1.29(d,J=6.8Hz,6H).

[0862] Example 22: N-(4-(4-amino-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 23)

[0863]

[0864] Compound T10 (90 mg, 0.288 mmol), 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15) (82 mg, 0.208 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (167 mg, 0.432 mmol), and N,N-diisopropylethylamine (113 mg, 0.865 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 23 (62 mg).

[0865] MS m / z (ESI): 556.1 [M+H] + .

[0866] 1H NMR (400MHz, DMSO-d6) δ11.64(s,1H),8.69(d,J=3.2Hz,1H),8.34(s,1H),8.28(s,1H ),8.09-8.03(m,1H),7.91(d,J=8.8Hz,2H),7.84(dd,J=8.8,4.0Hz,1H),7.71(d,J=8. 4Hz,2H),5.57-5.49(m,1H),4.11(dt,J=15.6,7.2Hz,2H),3.94(ddd,J=21.6,12.0,5 .6Hz,2H),3.11(dq,J=13.6,6.8Hz,1H),2.43(q,J=6.8Hz,2H),1.28(d,J=6.8Hz,6H).

[0867] Example 23: N-(4-(4-amino-1-(tetrahydrofuran-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 24)

[0868]

[0869] Compound T10 (34 mg, 0.10 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (33 mg, 0.10 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (47 mg, 0.50 mmol), and diisopropylethylamine (29 mg, 0.22 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 24 (2 mg).

[0870] MS m / z (ESI): 572.2 [M+H] + .

[0871] 1H NMR (400MHz, DMSO-d6) δ11.57 (s, 1H), 8.73 (d, J = 2.4Hz, 1H), 8.30 (s, 1H), 8.2 4(d,J=1.6Hz,2H),7.87(d,J=8.6Hz,2H),7.78(d,J=8.4Hz,1H),7.67(d,J=8. 4Hz,2H),5.53-5.44(m,1H),4.08(dt,J=15.8,7.3Hz,2H),3.90(ddd,J=21.6, 12.0,5.8Hz,2H),3.13-3.05(m,1H),2.43-2.32(m,2H),1.24(d,J=6.8Hz,6H).

[0872] Example 24: N-(4-(4-amino-1-isobutyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 25)

[0873]

[0874] Compound T20 (29 mg, 0.09 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (30 mg, 0.09 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (43 mg, 0.11 mmol), and diisopropylethylamine (29 mg, 0.22 mmol) were added to N,N-dimethylformamide (10 mL), and the reaction was carried out at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 25 (2 mg).

[0875] MS m / z (ESI): 559.1 [M+H] + .

[0876] 1H NMR (400MHz, DMSO-d6) δ11.60(s,1H),8.77(d,J=2.4Hz,1H),8.34(s,1H),8.27(dd,J=5.6,3.2Hz,2H),7.91(d,J=8.8Hz,2H),7.8 2(d,J=8.8Hz,1H),7.71(d,J=8.8Hz,2H),4.18(d,J=7.2Hz,2H),3.18-3.07(m,1H),2.32-2.22(m,1H),1.29(s,6H),0.91(s,6H).

[0877] Example 25: N-(4-(4-amino-1-isobutyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 26)

[0878]

[0879] Compound T20 (63 mg, 0.212 mmol), 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15) (62 mg, 0.212 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (124 mg, 0.318 mmol), and N,N-diisopropylethylamine (84 mg, 0.637 mmol) were added to N,N-dimethylformamide (5 mL), and the mixture was reacted at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 26 (15 mg).

[0880] MS m / z (ESI): 542.2 [M+H] + .

[0881] 1H NMR (400MHz, DMSO-d6) δ11.65 (s, 1H), 8.71 (d, J = 3.0Hz, 1H), 8.35 (s, 1H), 8. 28(s,1H),8.08(td,J=8.4,3.0Hz,1H),7.92(d,J=8.6Hz,2H),7.86(dd,J=8. 8,4.1Hz,1H),7.72(d,J=8.6Hz,2H),4.19(d,J=7.2Hz,2H),3.17-3.09(m,1H ), 2.30(dt,J=13.6,6.8Hz,1H), 1.29(d,J=6.9Hz,6H), 0.91(d,J=6.7Hz,6H).

[0882] Example 26: N-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 27)

[0883]

[0884] Compound T23 (55 mg, 0.18 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (61 mg, 0.18 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (143 mg, 0.37 mmol), and diisopropylethylamine (72 mg, 0.73 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 27 (2 mg).

[0885] MS m / z (ESI): 545.3 [M+H] + .

[0886] 1H NMR (400MHz, DMSO-d6) δ11.60(s,1H),8.77(d,J=2.4Hz,1H),8.34(s,1H),8.25-8.28(m,3H),7.92(s,1H),7.90(s,1H),7.83(d, J=8.4Hz,1H),7.72(d,J=8.4Hz,2H),5.18-5.05(m,1H),3.17-3.09(m,1H),1.53(s,3H),1.51(s,3H),1.30(s,3H),1.28(s,3H).

[0887] Example 27: N-[4-(4-amino-1-isopropyl-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl]-2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 28)

[0888]

[0889] Compound T23 (80 mg, 283 μmol) and 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14) (87 mg, 283 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 340 μmol) and diisopropylethylamine (75 mg, 567 μmol) were added, and the mixture was stirred at 25 °C for 5 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 28 (56 mg).

[0890] MS m / z (ESI): 527.2 [M+H] + .

[0891] 1 H NMR(400MHz,DMSO-d6)δ11.80(s,1H),8.32(s,1H),8.26(s,1H),7.93-7.91(m,2H),7.74-7.70(m,4H), 7.45-7.41(m,2H),5.12-5.06(m,1H),3.17-3.10(m,1H),1.52(d,J=8.0Hz,6H),1.28(d,J=8.0Hz,6H).

[0892] Example 28: N-(4-(4-amino-1-(2-fluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 29)

[0893]

[0894] Compound T22 (51 mg, 0.17 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (56 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (72 mg, 0.19 mmol), and diisopropylethylamine (67 mg, 0.51 mmol) were added to N,N-dimethylformamide (10 mL), and the reaction was carried out at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 29 (15 mg).

[0895] MS m / z (ESI): 548.2 [M+H] + .

[0896] 1 H NMR (400MHz, DMSO-d6) δ11.60 (s, 1H), 8.77 (d, J = 2.4Hz, 1H), 8.34 (s, 1H), 8.29 ( s,1H),8.28-8.24(m,1H),7.93(s,1H),7.91(s,1H),7.84(s,1H),7.82(s,1H),7 .74(s,1H),7.72(s,1H),4.97(t,J=4.8Hz,1H),4.86(t,J=4.8Hz,1H),4.72(t,J =4.8Hz,1H),4.65(t,J=4.8Hz,1H),3.11-3.17(m,1H),1.30(s,3H),1.28(s,3H).

[0897] Example 29: N-(4-(4-amino-1-(oxacyclobut-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 30)

[0898]

[0899] Compound T9 (53 mg, 0.17 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (56 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (72 mg, 0.19 mmol), and diisopropylethylamine (67 mg, 0.51 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 30 (10 mg).

[0900] MS m / z (ESI): 558.2 [M+H] + .

[0901] 1 H NMR (400MHz, DMSO-d6) δ11.62(s,1H),8.77(d,J=2.4Hz,1H),8.35(s,1H),8.28(s,1H),8.26(d,J=2.4Hz,1H),7.94(d,J=8.8Hz,2H),7.82-7.84 (m,1H),7.78(d,J=8.8Hz,2H),6.09-5.97(m,1H),5.13(t,J=6.4Hz,2H) ,5.01(d,J=6.4Hz,2H),3.14(p,J=6.8Hz,1H),1.30(s,3H),1.28(s,3H).

[0902] Example 30: N-(4-(4-amino-(2-fluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 31)

[0903]

[0904] Compound T22 (95 mg, 331 μmol) and 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14) (96 mg, 331 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (251 mg, 661 μmol) and diisopropylethylamine (128 mg, 992 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by column chromatography to obtain the title compound 31 (56 mg).

[0905] MS m / z (ESI): 531.2 [M+H] + .

[0906] 1 H NMR(400MHz,DMSO-d6)δ11.79(s,1H),8.31(s,1H),8.29(s,1H),7.93-7.91(m,2H),7.74-7.71(m,4H),7.45-7.40(m,2H),4.93 (t,J=4.8Hz,1H),4.86(t,J=4.8Hz,1H),4.72(t,J=4.8Hz,1H),4.65(t,J=4.8Hz,1H),3.19-3.08(m,1H),1.29(d,J=6.8Hz,6H).

[0907] Example 31: N-(4-(4-amino-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 32)

[0908]

[0909] Compound T6 (60 mg, 0.185 mmol), 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15) (54 mg, 0.185 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (95 mg, 0.243 mmol), and N,N-diisopropylethylamine (73 mg, 0.555 mmol) were added to N,N-dimethylformamide (5 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 32 (25 mg).

[0910] MS m / z (ESI): 570.2 [M+H] + .

[0911] 1 H NMR(400MHz,DMSO-d6)δ11.58(s,1H),8.64(d,J=3.2Hz,1H),8.28(s,1H),8.21(s,1H),8.01 (td,J=8.4,3.2Hz,1H),7.85(d,J=8.4Hz,2H),7.79(dd,J=8.8,4.0Hz,1H),7.65(d,J=8.8Hz ,2H),4.96-4.84(m,1H),3.97(dd,J=11.2,3.6Hz,2H),3.51(t,J=11.2Hz,2H),3.07(dt,J=1 3.6, 6.8Hz, 1H), 2.17 (qd, J = 12.4, 4.4Hz, 2H), 1.85 (d, J = 10.4Hz, 2H), 1.22 (d, J = 6.8Hz, 6H).

[0912] Example 32: N-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 33)

[0913]

[0914] Compound T23 (52 mg, 0.185 mmol), 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15) (54 mg, 0.185 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (95 mg, 0.243 mmol), and N,N-diisopropylethylamine (73 mg, 0.555 mmol) were added to N,N-dimethylformamide (5 mL), and the mixture was reacted at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 33 (8 mg).

[0915] MS m / z (ESI): 528.2 [M+H] + .

[0916] 1 H NMR (400MHz, DMSO-d6) δ11.64(s,1H),8.70(d,J=3.2Hz,1H),8.35(s,1H),8.26(s,1H),8.07(td,J=8.4,3.2Hz,1H),7.92(d,J=8.4Hz,2H),7.8 5(dd,J=8.8,4.0Hz,1H),7.71(d,J=8.8Hz,2H),5.14-5.04(m,1H),3.14(dt,J=13.6,6.8Hz,1H),1.52(d,J=6.8Hz,6H),1.29(d,J=6.8Hz,6H).

[0917] Example 33: N-(4-(4-amino-1-(2-fluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 34)

[0918]

[0919] Compound T22 (40 mg, 0.136 mmol), 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17) (50 mg, 0.136 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (80 mg, 0.205 mmol), and N,N-diisopropylethylamine (54 mg, 0.410 mmol) were added to N,N-dimethylformamide (5 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was slurried with methanol and filtered to give the title compound 34 (40 mg).

[0920] MS m / z (ESI): 547.1 [M+H] + .

[0921] 1 H NMR (400MHz, DMSO-d6) δ11.75(s,1H),8.30(d,J=6.8Hz,2H),7.93(d,J=8.4Hz,2H),7.76-7.72(m,4H),7.69-7.64(m,2H),4.98 (t,J=4.8Hz,1H),4.86(t,J=4.8Hz,1H),4.72(t,J=4.8Hz,1H),4.65(t,J=4.8Hz,1H),3.18-3.11(m,1H),1.30(d,J=6.8Hz,6H).

[0922] Example 34: N-(4-(4-amino-1-(2-fluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 35)

[0923]

[0924] Compound T22 (53 mg, 0.185 mmol), 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15) (54 mg, 0.185 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (95 mg, 0.243 mmol), and N,N-diisopropylethylamine (73 mg, 0.555 mmol) were added to N,N-dimethylformamide (5 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was slurried with methanol and filtered to give the title compound 35 (26 mg).

[0925] MS m / z(ESI): 532.1 [M+H] + .

[0926] 1 H NMR (400MHz, DMSO-d6) δ11.64(s,1H),8.69(d,J=3.2Hz,1H),8.33(s,1H),8.28(s, 1H),8.06(td,J=8.4,3.2Hz,1H),7.92(d,J=8.4Hz,2H),7.84(dd,J=8.8,4.0Hz,1H) ,7.72(d,J=8.8Hz,2H),4.97(t,J=4.8Hz,1H),4.85(t,J=4.8Hz,1H),4.71(t,J=4.8 Hz,1H),4.64(t,J=4.8Hz,1H),3.13(dt,J=13.6,6.8Hz,1H),1.28(d,J=6.8Hz,6H).

[0927] Example 35: N-(4-(4-amino-1-(oxacyclobut-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 36)

[0928]

[0929] Compound T9 (55 mg, 0.185 mmol), 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17) (57 mg, 0.185 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (108 mg, 0.277 mmol), and N,N-diisopropylethylamine (73 mg, 0.555 mmol) were added to N,N-dimethylformamide (5 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was slurried with methanol and filtered to give the title compound 36 (20 mg).

[0930] MS m / z (ESI): 557.2 [M+H] + .

[0931] 1H NMR (400MHz, DMSO-d6) δ11.76(s,1H),8.32(s,1H),8.28(s,1H),7.95(d,J=8.4Hz,2H),7.77(dd,J=8.4,4.0Hz,2H),7.76-7.72(m, 2H),7.68-7.65(m,2H),6.09-6.00(m,1H),5.13(t,J=6.4Hz,2H),5.03(t,J=7.2Hz,2H),3.19-3.10(m,1H),1.30(d,J=6.4Hz,6H).

[0932] Example 36: N-(4-(4-amino-1-(2,2,2-trifluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 37)

[0933]

[0934] Compound T24 (58 mg, 0.17 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (56 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (72 mg, 0.19 mmol), and diisopropylethylamine (67 mg, 0.51 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 37 (10 mg).

[0935] MS m / z (ESI): 584.2 [M+H] + .

[0936] 1 H NMR (400MHz, DMSO-d6) δ11.62(s,1H),8.78(d,J=2.4Hz,1H),8.35(d,J=3.2Hz,2H),8.28(dd,J=8.4,2.4Hz,1H),7.94(d, J=8.4Hz,2H),7.83(d,J=8.4Hz,1H),7.74(d,J=8.4Hz,2H),5.30(q,J=9.2Hz,2H),3.15(m,1H),1.31(s,3H),1.29(s,3H).

[0937] Example 37: N-(4-(4-amino-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 38)

[0938]

[0939] Compound T6 (59 mg, 0.17 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (56 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (72 mg, 0.19 mmol), and diisopropylethylamine (67 mg, 0.51 mmol) were added to N,N-dimethylformamide (10 mL), and the reaction was carried out at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 38 (20 mg).

[0940] MS m / z (ESI): 586.2 [M+H] + .

[0941] 1 H NMR (400MHz, DMSO-d6) δ11.60 (s, 1H), 8.77 (d, J = 2.4Hz, 1H), 8.34 (s, 1H), 8.2 5-8.29(m,2H),7.92(d,J=8.8Hz,2H),7.83(d,J=8.4Hz,1H),7.72(d,J=8.4Hz ,2H),4.94-5.00(m,1H),4.02-4.05(m,2H),3.58(t,J=11.2Hz,2H),3.11-3.1 7(m,1H),2.18-2.24(m,2H),1.92(d,J=10.4Hz,2H),1.30(s,3H),1.28(s,3H).

[0942] Example 38: N-(4-(4-amino-7-(2,2-difluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 39)

[0943]

[0944] Compound T3 (63 mg, 0.170 mmol), 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17) (56 mg, 0.170 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (86 mg, 0.222 mmol), and N,N-diisopropylethylamine (68 mg, 0.512 mmol) were added to N,N-dimethylformamide (5 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was slurried with methanol and filtered to give the title compound 39 (28 mg).

[0945] MS m / z (ESI): 564.1 [M+H] + .

[0946] 1 H NMR (400MHz, DMSO-d6) δ11.69(s,1H),8.31(s,1H),8.21(s,1H),7.86(d,J=8.4Hz,2H),7.73(d,J=8.8Hz,2H),7.66(d,J=8.8Hz,2H),7.52(d ,J=8.4Hz,2H),7.38(s,1H),6.59(s,1H),6.46(s,1H),6.32(s,1H),4.74-4.61(m,2H),3.14(dt,J=13.6,6.8Hz,1H),1.30(d,J=6.8Hz,6H).

[0947] Example 39: N-(4-(4-amino-7-(2-fluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 40)

[0948]

[0949] Compound T25 (64 mg, 0.184 mmol), 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17) (60 mg, 0.184 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (107 mg, 0.276 mmol), and N,N-diisopropylethylamine (73 mg, 0.553 mmol) were added to N,N-dimethylformamide (5 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was slurried with methanol and filtered to give the title compound 40 (21 mg).

[0950] MS m / z (ESI): 546.1 [M+H] + .

[0951] 1 H NMR (400MHz, DMSO-d6) δ11.68(s,1H),8.31(s,1H),8.19(s,1H),7.85(d,J=8. 4Hz,2H),7.73(d,J=8.8Hz,2H),7.66(d,J=8.8Hz,2H),7.52(d,J=8.4Hz,2H), 7.41(s,1H),4.89(t,J=4.8Hz,1H),4.77(t,J=4.8Hz,1H),4.56(t,J=4.8Hz,1 H), 4.49 (t, J = 4.8Hz, 1H), 3.14 (dt, J = 13.6, 6.8Hz, 1H), 1.30 (d, J = 6.8Hz, 6H).

[0952] Example 40: N-[4-[4-amino-7-(2,2-difluoroethyl)-1H-pyrrolo[3,4-d]pyrimidin-5-yl]phenyl]-2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 41)

[0953]

[0954] Weigh out T21 (100 mg, 277 μmol) and 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14) (802 mg, 277 μmol), dissolve in N,N-dimethylformamide (2 mL), add triethylamine (87 mg, 630 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (130 mg, 332 μmol), and stir at 25 °C for 2 hours. Add water to the reaction solution, extract with ethyl acetate, and combine the organic phases. Wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and separate the crude product by preparative high performance liquid chromatography to obtain the title compound 41 (49 mg).

[0955] MS m / z (ESI): 548.2 [M+H] + .

[0956] 1H NMR (400MHz, DMSO-d6) δ11.73(s,1H),8.31(s,1H),8.20(s,1H),7.86(d,J=8.0Hz,2H),7.74-7.71(m,2H),7.51(d ,J=8.0Hz,2H),7.44-7.38(m,3H),6.60-6.31(m,1H),4.72-4.63(m,2H),3.17-3.11(m,1H),1.29(d,J=7.2Hz,6H).

[0957] Example 41: N-(4-(4-amino-7-(2,2-difluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 42)

[0958]

[0959] Compound T3 (63 mg, 0.17 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (56 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (72 mg, 0.19 mmol), and diisopropylethylamine (67 mg, 0.51 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 42 (20 mg).

[0960] MS m / z (ESI): 565.2 [M+H] + .

[0961] 1 H NMR (400MHz, DMSO-d6) δ11.56(s,1H),8.77(d,J=2.4Hz,1H),8.34(s,1H),8.27(dd,J=8.4,2.4Hz,1H),8.23(s,1H),7.84(dd,J=14.4,8.4Hz ,3H),7.60(s,1H),7.55(d,J=8.4Hz,2H),5.70(dd,J=14.8,7.2Hz,1H),3.10-3.15(m,1H),1.77(d,J=7.2Hz,2H),1.30(s,3H),1.28(s,3H).

[0962] Example 42: N-(4-(4-amino-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 43)

[0963]

[0964] Compound T5 (78 mg, 0.23 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (74 mg, 0.23 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (177 mg, 0.46 mmol), and diisopropylethylamine (150 mg, 1.2 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 43 (27 mg).

[0965] MS m / z (ESI): 583.2 [M+H] + .

[0966] 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),8.77(d,J=2.4Hz,1H),8.34(s,1H),8.27(dd,J=8.4,2.4Hz,1H),8.18(s,1H),7.84(t,J=9. 2Hz,4H),7.51(d,J=8.4Hz,2H),7.40(s,1H),4.76-4.90(m,2H),4.43-4.57(m,2H),3.10-3.17(m,1H),1.29(s,3H),1.28(s,3H).

[0967] Example 43: N-(4-(4-amino-7-(1,1,1-trifluoropropane-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 44)

[0968]

[0969] Compound T1 (89 mg, 0.22 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (70 mg, 0.22 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (167 mg, 0.43 mmol), and diisopropylethylamine (85 mg, 0.64 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 44 (27 mg).

[0970] MS m / z (ESI): 597.2 [M+H] + .

[0971] 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),8.77(d,J=2.4Hz,1H),8.34(s,1H),8.27(dd,J=8.4,2.4Hz,1H),8.21(s,1H),7.7 8-7.84(m,3H),7.49(s,2H),7.47(s,1H),7.35(s,1H),4.61-4.68(m,2H),3.07-3.14(m,1H),1.29(s,3H),1.28(s,3H).

[0972] Example 44: N-(4-(4-amino-7-(2-fluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 45)

[0973]

[0974] Compound T25 (22 mg, 0.062 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (20 mg, 0.062 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (48 mg, 0.12 mmol), and diisopropylethylamine (24 mg, 0.18 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 45 (15 mg).

[0975] MS m / z (ESI): 547.2 [M+H] + .

[0976] 1 H NMR (400MHz, DMSO-d6) δ11.55(s,1H),8.78(d,J=2.4Hz,1H),8.34(s,1H),8.27(dd,J=8.4,2.4Hz,1H),8.24(s,1H),7.86(d,J=8.4Hz ,2H),7.83(d,J=8.4Hz,1H),7.52(d,J=8.4Hz,2H),7.40(s,1H),5.11(t,J=9.6Hz,2H),3.21-3.03(m,1H),1.30(s,3H),1.28(s,3H).

[0977] Example 45: N-(4-(4-amino-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 46)

[0978]

[0979] Compound T7 (75 mg, 0.17 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (56 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (130 mg, 0.34 mmol), and diisopropylethylamine (72 mg, 0.54 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 46 (15 mg).

[0980] MS m / z (ESI): 585.2 [M+H] + .

[0981] 1H NMR (400MHz, DMSO-d6) δ11.54(s,1H),8.77(d,J=2.4Hz,1H),8.34(s,1H),8.28(d,J=8.4Hz,1H),8.17(s,1H),7.84(d,J=7.2Hz,3H),7.53(s,3H ),4.87(s,1H),4.03(d,J=7.8Hz,2H),3.56(t,J=12.0Hz,2H),3.19-3.0 8(m,1H),2.15(d,J=12.0Hz,2H),1.91(m,2H),1.30(s,3H),1.28(s,3H).

[0982] Example 46: N-(4-(4-amino-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 47)

[0983]

[0984] Compound T21 (93 mg, 321 μmol) and 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (88 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 47 (52 mg).

[0985] MS m / z (ESI): 546.2 [M+H] + .

[0986] 1 H NMR (400MHz, DMSO-d6) δ11.60(s,1H),8.71(d,J=3.2Hz,1H),8.35(s,1H),8.23(s,1H),8.09(m,1H),7.86( m,3H),7.61(s,1H),7.55(d,J=8.4Hz,2H),3.22-3.10(m,1H),1.78(d,J=7.2Hz,3H),1.29(d,J=6.8Hz,6H).

[0987] Example 47: N-(4-(4-amino-7-(1,1,1-trifluoropropane-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 48)

[0988]

[0989] Compound T1 (103 mg, 321 μmol) and 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (88 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 48 (78 mg).

[0990] MS m / z (ESI): 577.2 [M+H] + .

[0991] 1 H NMR (400MHz, DMSO-d6) δ11.59(s,1H),8.71(d,J=3.2Hz,1H),8.35(s,1H),8.21(s,1H),8.09( m,1H),7.85(m,3H),7.51(d,J=8.4Hz,2H),7.37(s,1H),4.73-4.86(m,1H),4.46-4.52(m,1H), 3.09-3.12(m,1H),1.29(d,J=6.8Hz,6H).

[0992] Example 48: N-(4-(4-amino-7-(1,1,1-trifluoropropane-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 49)

[0993]

[0994] Compound T1 (55 mg, 0.17 mmol), 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15) (54 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (130 mg, 0.34 mmol), and diisopropylethylamine (85 mg, 0.64 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 49 (31 mg).

[0995] MS m / z (ESI): 581.2 [M+H] + .

[0996] 1 H NMR (400MHz, DMSO-d6) δ11.59 (s, 1H), 8.71 (d, J = 2.4Hz, 1H), 8.35 (m, 1H), 8.21 (m, 1H), 8.07-8.09 (m,1H),7.85(m,3H),7.51(m,2H),7.38(s,1H),4.69-4.66(m,2H),3.14-3.11(m,1H),1.29(m,6H).

[0997] Example 49: N-(4-(4-amino-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 50)

[0998]

[0999] Compound T2 (78 mg, 0.17 mmol), 2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (56 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (130 mg, 0.34 mmol), and diisopropylethylamine (85 mg, 0.64 mmol) were added to N,N-dimethylformamide (10 mL), and the mixture was reacted at 26 °C for 12 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was separated by preparative high-performance liquid chromatography to obtain the title compound 50 (27 mg).

[1000] MS m / z (ESI): 598.2 [M+H]+ .

[1001] 1 H NMR (400MHz, DMSO-d6) δ11.63(s,1H),8.78(d,J=2.4Hz,1H),8.35(d,J=4.8Hz,2H),8.28(dd,J=8.4,2.4Hz,1H),7.94(d,J=8.4Hz,2H) ,7.83(d,J=8.4Hz,1H),7.73(d,J=8.4Hz,2H),5.81-5.68(m,1H),3.18-3.07(m,1H),1.81(d,J=7.2Hz,3H),1.30(s,3H),1.28(s,3H).

[1002] Example 50: N-[4-[4-amino-7-(2-fluoroethyl)pyrrolo[2,3-d]pyrimidin-5-yl]phenyl]-2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 51)

[1003]

[1004] Weigh out T25 (100 mg, 332 μmol) and 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14) (96 mg, 332 μmol), dissolve in N,N-dimethylformamide (4 mL), add triethylamine (104 mg, 995 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (156 mg, 398 μmol), and stir at 25 °C for 2 hours. Add water to the reaction solution, extract with ethyl acetate, and combine the organic phases. Wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and separate the crude product by preparative high performance liquid chromatography to obtain the title compound 51 (36 mg).

[1005] MS m / z (ESI): 530.2 [M+H] + .

[1006] 1H NMR (400MHz, DMSO-d6) δ11.73(s,1H),8.31(s,1H),8.29(s,1H),7.87-7.85(m,2H),7.74-7.71(m,2H),7.51(d,J=8.0Hz,2H),7.45-7.41(m ,3H),4.89(t,J=4.8Hz,1H),4.77(t,J=4.8Hz,1H),4.55(t,J=4.8Hz,1H),4.49(t,J=4.8Hz,1H),3.17-3.08(m,1H),1.29(d,J=6.8Hz,6H).

[1007] Example 51: N-[4-[4-amino-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl]phenyl]-2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 52)

[1008]

[1009] Weigh out T5 (105 mg, 325 μmol) and 2-(4-fluorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T14) (76 mg, 260 μmol), dissolve in N,N-dimethylformamide (4 mL), add triethylamine (99 mg, 976 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (151 mg, 390 μmol), and stir at 25 °C for 2 hours. Add water to the reaction solution, extract with ethyl acetate, and combine the organic phases. Wash with saturated brine, dry with anhydrous sodium sulfate, concentrate, and separate the crude product by preparative high performance liquid chromatography to obtain the title compound 52 (57 mg).

[1010] MS m / z (ESI): 566.2 [M+H] + .

[1011] 1 H NMR(400MHz,DMSO-d6)δ11.75(s,1H),8.31(s,1H),8.23(s,1H),7.87-7.85(m,2H),7.74-7.71(m,2H), 7.51(d,J=8.0Hz,2H),7.45-7.41(m,3H),5.15-5.10(m,2H),3.17-3.08(m,1H),1.29(d,J=6.8Hz,6H).

[1012] Example 52: N-(4-(4-amino-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 53)

[1013]

[1014] Compound T7 (136 mg, 0.307 mmol), 2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17) (100 mg, 0.307 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (179 mg, 0.461 mmol), and N,N-diisopropylethylamine (121 mg, 0.922 mmol) were added to N,N-dimethylformamide (10 mL), and the reaction was carried out at 25 °C for 4 hours. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the concentrated residue was slurried with methanol and filtered to give the title compound 53 (12 mg).

[1015] MS m / z (ESI): 584.2 [M+H] + .

[1016] 1 H NMR (400MHz, DMSO-d6) δ11.68(s,1H),8.30(s,1H),8.17(s,1H),7.85(d,J=8. 4Hz,2H),7.72(d,J=8.8Hz,2H),7.66(d,J=8.8Hz,2H),7.52(d,J=7.6Hz,3H),4 .86(s,1H),4.03(d,J=8.4Hz,2H),3.56(t,J=11.6Hz,2H),3.14(dd,J=13.6,6 .8Hz,1H),2.14(d,J=8.4Hz,2H),1.90(d,J=9.6Hz,2H),1.29(d,J=6.8Hz,6H).

[1017] Example 53: N-(4-(4-amino-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 54)

[1018]

[1019] Compound T5 (99 mg, 321 μmol) and 6-isopropyl-2-(5-methylpyridin-2-yl)-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T13) (88 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 54 (67 mg).

[1020] MS m / z (ESI): 563.2 [M+H] + .

[1021] 1 H NMR (400MHz, DMSO-d6) δ11.60 (s, 1H), 8.71 (d, J = 3.2Hz, 1H), 8.35 (s, 1H), 8.24 (s, 1H), 8.08 (m, 1H), 7.92-7.8 1(m,3H),7.52(d,J=8.4Hz,2H),7.41(s,1H),5.13(q,J=9.2Hz,2H),3.20-3.04(m,1H),1.29(d,J=6.8Hz,6H).

[1022] Example 54: N-(4-(4-amino-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 55)

[1023]

[1024] Compound T7 (99 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15) (88 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 55 (58 mg).

[1025] MS m / z (ESI): 569.2 [M+H] + .

[1026] 1 H NMR (400MHz, DMSO-d6) δ11.59(s,1H),8.71(d,J=3.2Hz,1H),8.35(s,1H),8.17(s,1H),8.09(m,1H),7.86(m,1H),7.81-7.84(m,3H),7.48-7. 51(m,3H),4.87(m,1H),4.05(m,2H),3.56(t,J=11.6Hz,2H),3.21-3.0 4(m,1H),2.21-2.07(m,2H),1.95-1.82(m,2H),1.29(d,J=6.8Hz,6H).

[1027] Example 55: N-(4-(4-amino-7-(oxacyclobut-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 56)

[1028]

[1029] Compound T27 (90 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (94 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 56 (11 mg).

[1030] MS m / z (ESI): 557.2 [M+H] + .

[1031] 1H NMR (400MHz, DMSO-d6) δ11.57(s,1H),8.78(d,J=2.4Hz,1H),8.35(s,1H),8.28(dd,J=8.4,2.4Hz,1H),8.18(s,1H),7.87(d,J=8.4Hz,2H),7.84- 7.78(m,2H),7.57(d,J=8.8Hz,2H),5.91(t,J=7.2Hz,1H),5.05(t,J=6.4 Hz,2H),5.01(d,J=7.2Hz,2H),3.17-3.10(m,1H),1.29(d,J=6.4Hz,6H).

[1032] Example 56: N-(4-(4-amino-7-(2,2-difluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-3-fluorophenyl)-2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 57)

[1033]

[1034] Compound T28 (98 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17) (93 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 57 (10 mg).

[1035] MS m / z (ESI): 582.2 [M+H] + .

[1036] 1 H NMR (400MHz, DMSO-d6) δ11.75(s,1H),8.28(s,1H),8.17(s,1H),7.89-7.92(m,1H),7.62-7.71(m,4H),7.50-7.52( m,1H),7.40-7.44(m,1H),7.37(s,1H),6.29-6.57(m,1H),4.63-4.70(m,2H),3.07-3.14(m,1H),1.25-1.27(m,6H).

[1037] Example 57: N-(4-(4-amino-7-(oxacyclobut-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 58)

[1038]

[1039] Compound T28 (98 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17) (93 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 58 (9 mg).

[1040] MS m / z (ESI): 556.2 [M+H] + .

[1041] 1 H NMR (400MHz, DMSO-d6) δ11.68(s,1H),8.29(s,1H),8.15(s,1H),7.83-7.85(m,2H),7.76(s,1H),7.62-7. 72(m,4H),7.52-7.55(m,2H),5.84-5.91(m,1H),4.96-5.04(m,4H),3.07-3.14(m,1H),1.26-1.27(m,6H).

[1042] Example 58: N-(4-(4-amino-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-2-(5-chloropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 59)

[1043]

[1044] Compound T29 (104 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T18) (94 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 59 (31 mg).

[1045] MS m / z (ESI): 601.2 [M+H] + .

[1046] 1 H NMR (400MHz, DMSO-d6) δ11.97(s,1H),8.78(s,1H),8.57(t,J=8.0Hz,1H),8.42(s,1H),8.27-8.30(m,1H),8. 24(s,1H),7.83(d,J=8.0Hz,1H),7.38-7.48(m,3H),5.09-5.06(m,2H),3.13-3.20(m,1H),1.28-1.30(m,6H).

[1047] Example 59: N-(4-(4-amino-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-fluoropyridin-2-yl)-6-(2-methoxyethyl)-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 60)

[1048]

[1049] Compound T5 (98 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T30) (94 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 60 (4 mg).

[1050] MS m / z (ESI): 583.2 [M+H] + .

[1051] 1 H NMR (400MHz, DMSO-d6) δ11.59(s,1H),8.71(d,J=2.8Hz,1H),8.34(s,1H),8.23(s,1H),8.09(td,J=8.4,2.8Hz,1H), 7.87(d,J=8.4Hz,3H),7.52(d,J=8.4Hz,2H),7.41(s,1H),3.71(t,J=6.4Hz,2H),3.30(s,3H),3.03(t,J=6.4Hz,2H).

[1052] Example 60: N-(4-(4-amino-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(4-chlorophenyl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 61)

[1053]

[1054] Compound T21 (93 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T17) (94 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 61 (7 mg).

[1055] MS m / z (ESI): 565.2 [M+H] + .

[1056] 1H NMR(400MHz,DMSO-d6)δ11.75(s,1H),8.28-8.29(m,2H),7.89-7.91(m,2H),7.68-7.71(m,4H), 7.62-7.66(m,2H),6.34-6.63(m,1H),4.76-4.84(m,2H),3.07-3.14(m,1H),1.25-1.27(m,6H).

[1057] Example 61: N-(4-(4-amino-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(4-chlorophenyl)-6-cyclopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 62)

[1058]

[1059] Compound T5 (98 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T31) (93 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 62 (3 mg).

[1060] MS m / z (ESI): 580.2 [M+H] + .

[1061] 1 H NMR (400MHz, DMSO-d6) δ11.71(s,1H),8.28(s,1H),8.16(s,1H),7.83-7.86(m,2H),7.61-7.67(m,4H),7. 48-7.51(m,2H),7.46(s,1H),5.10-5.17(m,2H),2.20-2.27(m,1H),1.03-1.05(m,2H),0.92-0.94(m,2H).

[1062] Example 62: N-(4-(4-amino-7-(oxacyclobut-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-fluoropyrimidin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 63)

[1063]

[1064] Compound T27 (90 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15) (89 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 63 (21 mg).

[1065] MS m / z (ESI): 541.2 [M+H] + .

[1066] 1 H NMR (400MHz, DMSO-d6) δ11.60(s,1H),8.71-8.72(m,1H),8.35(s,1H),8.18(s,1H),8.06-8.11(m,1H),7.84-7.88( m,3H),7.79(s,1H),7.55-7.57(m,2H),5.87-5.94(m,1H),4.99-5.07(m,4H),3.01-3.17(m,1H),1.28-1.29(m,6H).

[1067] Example 63: N-(4-(4-amino-7-(2,2,2-trifluoroethyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-2-(5-chloropyrimidin-2-yl)-6-cyclopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 64)

[1068]

[1069] Compound T5 (98 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T32) (93 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 64 (15 mg).

[1070] MS m / z (ESI): 581.2 [M+H] + .

[1071] 1 H NMR(400MHz,DMSO-d6)δ11.54(s,1H),8.73(s,1H),8.17-8.25(m,3H),7.76-7.84(m,3H),7.48-7.5 0(m,2H),7.37(s,1H),5.06-5.13(m,2H),2.20-2.26(m,1H),1.02-1.06(m,2H),0.89-0.93(m,2H).

[1072] Example 64: N-(4-(4-amino-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxamide (Compound 65)

[1073]

[1074] Compound T21 (93 mg, 321 μmol) and 2-(5-fluoropyridin-2-yl)-6-isopropyl-3-oxo-2,3-dihydropyridazine-4-carboxylic acid (T15) (89 mg, 321 μmol) were weighed and dissolved in N,N-dimethylformamide (4 mL). Diisopropylethylamine (85 mg, 641 μmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (149 mg, 384 μmol) were added, and the mixture was stirred at 25 °C for 2 hours. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by preparative high-performance liquid chromatography to obtain the title compound 65 (12 mg).

[1075] MS m / z (ESI): 550.2 [M+H] + .

[1076] 1 H NMR(400MHz,DMSO-d6)δ11.67(s,1H),8.71-8.72(m,1H),8.35(s,1H),8.32(s,1H),8.06-8.11(m,1 H),7.92-7.95(m,2H),7.84-7.87(m,1H),7.72-7.74(m,2H),6.37-6.66(m,1H),4.79-4.87(m,2H), 3.10-3.17(m,1H),1.28-1.30(m,6H).

[1077] Biological evaluation:

[1078] Experimental Example 1: Test of Compound Activity Against Axl Kinase

[1079] 1. Test system:

[1080] Enzyme: Axl (Carna)

[1081] HTRF kit: HTRF KinEASE TM -TK (Cisbio)

[1082] 2. Test parameters:

[1083] Enzyme (Axl) concentration: 1 nM;

[1084] Substrate (TK-Substrate-Biotin) concentration: 1 μM;

[1085] Streptavidin-XL665 concentration: 0.0625 μM;

[1086] TK Antibody-Cryptate concentration: 1×;

[1087] ATP: 21 μM

[1088] Pre-incubation time of compound and enzyme: 60 minutes at room temperature

[1089] Reaction time: 50 minutes at room temperature

[1090] Testing time: 60 minutes at room temperature

[1091] Microplate reader parameters: BMG PHERAstar HTRF

[1092] 3. Experimental methods:

[1093] Axl enzyme was pre-incubated with different concentrations of test compounds (500 nM, 50 nM, 5 nM) at 25 °C for 60 min. Then, 5×TK-substrate-biotin / ATP mixed working solution was added, and the reaction was incubated at 25 °C for 50 min. Finally, 2×Streptavidin-XL665 and TK Antibody-Cryptate mixed working solution was added, and the reaction was incubated at 25 °C for 60 min. The fluorescence signal ratio was then detected by a microplate reader.

[1094] 4. Data Processing:

[1095] Using the solvent group (DMSO) as a negative control and the reaction buffer group as a blank control, the percentage inhibition rate of different concentrations of the compound was calculated according to the following formula:

[1096] Percentage inhibition rate = (1 - (Ratio of different concentrations of compound - Ratio of blank control) / (Ratio of negative control - Ratio of blank control)) × 100%;

[1097] When the percentage inhibition rate is between 30% and 80%, the half-maximal inhibitory concentration (IC50) of the compound is calculated according to the following formula. 50 ) or range:

[1098] IC 50 =X×(1-percentage inhibition rate (%)) / percentage inhibition rate (%), where: X is the test concentration of the compound when the inhibition rate is between 30-80%.

[1099] 5. Experimental Results:

[1100] The inhibitory effect of the compound on Axl was determined according to the above method, and the results are shown in Table 1.

[1101] Table 1. Inhibitory activity of the compounds of the present invention against Axl

[1102] Compound numbering <![CDATA[IC 50 (nM)]]> 1 38.4 2 11.4 3 11.3 4 6.7 5 7.8 6 45.6 7 15.7 8 22.9 9 20.2 10 30.4 11 47.6 12-I 18.2 12-II 19.7 14 15.0 15 6.4 16 25.9 17 19.2 18 25.7

[1103] 19 14.6 20 16.8 21 37.8 22 36.2 23 12.5 24 8.5 25 13.9 26 21.0 27 4.4 28 15.0 29 4.7 30 21.8 31 7.4 32 13.2 33 2.1 34 19.1 35 11.9 36 33.7 37 4.4 38 3.8 39 28.1 40 15.3 41 19.7 42 30.4 43 15.3 44 30.9 45 9.0 46 5.0 47 9.6 48 9.9 49 28.2 50 37.2 51 7.6 52 44.7 53 24.1 54 9.1 55 6.1 56 31.3 57 8.7 58 31.1 59 12.9 60 5.2 61 11.0 62 16.9 63 20.8 64 10.2 65 6.9

[1104] in conclusion:

[1105] In the Axl Kinase Assay test, the compounds of this invention exhibited strong inhibitory activity.

[1106] Experimental Example 2: Test of the compound's activity against Mer, Tyro3, ​​and c-Met kinases

[1107] 1. Test system:

[1108] Kinase: Mer(Carna)

[1109] Kinase: Tyro3 (Carna)

[1110] Kinase: c-Met (Carna)

[1111] HTRF kit: HTRF KinEASE TM -TK (Cisbio)

[1112] 2. Test parameters:

[1113]

[1114] Pre-incubation time of compound and enzyme: 15 min / 15 min / 15 min at room temperature

[1115] Reaction time: 60 min / 30 min / 30 min at room temperature

[1116] Detection time: 60 min / 60 min / 60 min at room temperature

[1117] Microplate reader parameters: BMG PHERAstar HTRF

[1118] 3. Experimental Procedure:

[1119] The enzyme was pre-incubated with different concentrations of the test compounds (Mer: 500 nM, 50 nM, 5 nM; Tyro3 and c-Met: 1000 nM, 300 nM, 100 nM) at 25 °C for 15 min. Then, 5×TK-substrate-biotin / ATP mixed working solution was added, and incubation was carried out at 25 °C for a specified period (Mer: 60 min; Tyro3 and c-Met: 30 min). Finally, 2×Streptavidin-XL665 and TK Antibody-Cryptate mixed working solution was added, and after incubation at 25 °C for 1 h, the fluorescence signal ratio was detected using a microplate reader.

[1120] 4. Data Processing:

[1121] Using the solvent group (DMSO) as a negative control and the reaction buffer group as a blank control, the percentage inhibition rate of different concentrations of the compound was calculated according to the following formula:

[1122] Percentage inhibition rate = (1 - (Ratio of different concentrations of compound - Ratio of blank control) / (Ratio of negative control - Ratio of blank control)) × 100%;

[1123] When the percentage inhibition rate is between 30% and 80%, the half-maximal inhibitory concentration (IC50) of the compound is calculated according to the following formula. 50) or range:

[1124] IC 50 =X×(1-percentage inhibition rate (%)) / percentage inhibition rate (%), where: X is the test concentration of the compound when the inhibition rate is between 30-80%.

[1125] 5. Experimental Results:

[1126] The inhibitory effect of the compound on Mer / Tyro3 / c-Met was determined according to the above method, and the results are shown in Tables 2-1 to 2-3.

[1127] Table 2-1. Results of tests on the activity of compounds against Mer kinase

[1128] Compound numbering <![CDATA[Mer IC 50 (nM)]]> 2 92.0 3 94.9 4 73.5 7 93.2 14 90.2 15 120.6 16 113.8 17 97.0 18 99.9 29 52.0 32 159.4 34 182.6 35 111.1 38 108.0 39 126.4 40 81.7 43 97.4 44 122.1 45 103.8 46 121.4 47 80.1 48 112.6 49 63.3

[1129] 50 160.9 53 85.7

[1130] Table 2-2. Results of tests on the activity of compounds against Tyro3 kinase

[1131] Compound numbering <![CDATA[Tyro3 IC 50 (nM)]]> 2 900.0 3 1140.9 4 673.6 7 >1000 12-I 344.6 12-II 371.9 14 531.4 15 377.7 16 799.6 17 >1000 18 1193.9 29 299.6 32 904.7 34 544.6 35 424.9 38 255.5 39 266.8 40 1009.8 43 607.8 45 686.9 46 1488.4 47 >1000 48 681.9 49 876.6

[1132] Table 2-3. Results of tests on c-Met kinase activity of compounds

[1133] Compound numbering <![CDATA[c-Met IC 50 (nM)]]> 39 563.6 40 265.0 43 164.7 44 256.2 45 91.5 47 264.5 49 279.8

[1134] in conclusion:

[1135] Most of the compounds in this invention exhibit weak inhibitory activity against Mer, Tyro3, ​​and c-Met, indicating good selectivity relative to Axl.

[1136] Experimental Example 3: Assay on the inhibitory effect of the compound on the proliferation activity of Ba / F3-Axl cells

[1137] 1. Test system:

[1138] Cell Name / Manufacturer: Ba / F3-TEL-Axl / Hefei Puryson

[1139] Reagent name / manufacturer: Luminescent Cell Viability Assay,Promega

[1140] 2. Test parameters:

[1141] Cell count: 2000 cells / well

[1142] Culture medium: RPMI-1640 + 10% FBS + 1% P / S

[1143] DMSO content: 0.1%

[1144] Compound incubation conditions: 37℃, 5% CO2

[1145] Incubation time: 72 hours

[1146] Temperature detected: RT

[1147] Testing instruments: Molecular Devices

[1148] 3. Experimental Procedure:

[1149] Ba / F3-TEL-Axl cells were cultured in vitro as a monolayer in RPMI-1640 medium containing 10% FBS and 1% P / S at 37°C in an incubator with 5% CO2. Cells were seeded into 96-well plates at 2000 cells / well, and then a pre-diluted compound was added. DMSO was added to the negative control group, and culture medium was added to the blank control group. After incubation at 37°C in 5% CO2 for 72 hours, the CellTiter-Glo assay reagent was added to each well, and the relative chemiluminescence unit (RLU) value was read using a microplate reader in chemiluminescence detection mode.

[1150] 4. Data Processing:

[1151] Calculate the percentage inhibition rate of different concentrations of compounds using the following formula:

[1152] Percentage inhibition rate = (1 - (chemiluminescence signal value of test compound - chemiluminescence signal value of blank control) / (chemiluminescence signal value of negative control - chemiluminescence signal value of blank control)) × 100%. The half-maximal inhibitory concentration (IC50) of the compound was calculated by fitting a curve using Graphpad 7.0 with a four-parameter model. 50 ).

[1153] 5. Experimental Results:

[1154] The inhibitory activity of the compound on the proliferation of Ba / F3-TEL-Axl cells was determined according to the above method, and the results are shown in Table 3.

[1155] Table 3. Results of the inhibitory effect of the compounds on the proliferation activity of Ba / F3-TEL-Axl cells

[1156] Compound numbering <![CDATA[Ba / F3-TEL-Axl,IC 50 (nM)]]> 3 56.6 5 2.3 7 16.9 9 28.8 10 44.9 12-I 6.5 12-II 5.2 14 22.6 15 20.3 17 1.1 19 1.7 23 11.7 24 7.5 25 16.9 29 19.2 31 11.6 32 19.4 34 20.8 35 26.5 37 35.9 38 8.1 39 9.9 40 8.1 41 6.8 43 11.9 44 9.8 45 7.0

[1157] 46 3.5 47 9.2 48 7.9 49 5.7 50 22.6 51 8.3 53 21.4 54 44.6 55 9.6 57 16.0 58 44.6 60 18.0

[1158] in conclusion:

[1159] The compounds of this invention exhibit strong inhibitory activity against the proliferation of Ba / F3-TEL-Axl cells.

[1160] Experimental Example 4: CYP Enzyme (Cytochrome P450) Inhibition Assay

[1161] 1. Test system:

[1162] P450-Glo TM CYP1A2 screening system (Promega);

[1163] P450-Glo TM CYP2D6 screening system (Promega);

[1164] P450-Glo TM CYP3A4 screening system (Promega).

[1165] 2. Testing instruments:

[1166] BMG PHERAstar FS Luminescent.

[1167] 3. Test methods:

[1168] Perform the tests according to the kit instructions, as follows:

[1169] 3.1. Inhibition of CYP1A2:

[1170] Test group: Different concentrations of the analyte compound were added to microplates. Luciferin-ME (100 μM), K3PO4 (100 mM), and CYP1A2 (0.01 pmol / μL) were added to each well, and the mixture was pre-incubated at room temperature for 10 min. Then, an NADPH regeneration system was added, and the reaction was carried out at room temperature for 30 min. Finally, an equal volume of detection buffer was added, and the mixture was incubated at room temperature for 20 min. Chemiluminescence detection was then performed.

[1171] Negative control group: The experimental method was the same as that of the test group, but no test compound was added.

[1172] Blank control group: The experimental method is the same as that of the test group, but no test compound is added, and CYP1A2 Membrance (0.01 pmol / μL) is used instead of CYP1A2.

[1173] 3.2. Inhibition of CYP2D6:

[1174] Test group: Different concentrations of the analyte compounds were added to microplates. Luciferin-MEEGE (3 μM), K3PO4 (100 mM), and CYP2D6 (5 nM) were added to each well, and the mixture was pre-incubated at room temperature for 10 min. Then, an NADPH regeneration system was added, and the reaction was carried out at 37 °C for 30 min. Finally, an equal volume of detection buffer was added, and the mixture was incubated at room temperature for 20 min. Chemiluminescence detection was then performed.

[1175] Negative control group: The experimental method was the same as that of the test group, but no test compound was added.

[1176] Blank control group: The experimental method is the same as that of the test group, but no test compound is added, and CYP2D6 Membrance (5nM) is used instead of CYP2D6.

[1177] 3.3. Inhibition of CYP3A4:

[1178] Test group: Different concentrations of the analyte compounds were added to microplates. Luciferin-IPA (3 μM), K3PO4 (100 mM), and CYP3A4 (2 nM) were added to each well, and the plates were pre-incubated at room temperature for 10 min. Then, an NADPH regeneration system was added, and the reaction was carried out at room temperature for 30 min. Finally, an equal volume of detection buffer was added, and the plates were incubated at room temperature for 20 min. Chemiluminescence detection was then performed.

[1179] Negative control group: The experimental method was the same as that of the test group, but no test compound was added.

[1180] Blank control group: The experimental method is the same as that of the test group, but no test compound is added, and CYP3A4 Membrance (2nM) is used instead of CYP3A4.

[1181] 4. Data Processing:

[1182] Percentage inhibition rate (%) = (1 - (chemiluminescence signal value of the test compound - chemiluminescence signal value of the blank control) / (chemiluminescence signal value of the negative control - chemiluminescence signal value of the blank control)) × 100%;

[1183] Based on the inhibition rate of different concentrations of compounds on CYP enzymes, the half-maximal inhibitory concentration (IC50) of the compounds was estimated. 50 ) or range:

[1184] IC 50 =X×(1-percentage inhibition rate (%)) / percentage inhibition rate (%), where: X is the test concentration of the compound.

[1185] 5. Experimental Results:

[1186] The inhibition of the compound of the present invention on three CYPs was determined according to the above method, and the results are shown in Table 4 below.

[1187] Table 4. Results of CYPs Inhibition Test

[1188]

[1189] in conclusion:

[1190] The compounds of this invention do not show significant inhibitory effects on the three major CYP isoforms, indicating that their potential drug interaction is relatively low.

[1191] Experimental Example 5. Biochemical hERG Inhibition Assay

[1192] 1. Test system:

[1193] Reagent kit: Predictor TM hERG Fluorescence Polarization Assay,(ThermoFisher),

[1194] The kit contains:

[1195] Positive control compound: hERG potassium channel blocker E4031;

[1196] hERG cell membrane;

[1197] Tracer, an affinity tracer;

[1198] hERG buffer.

[1199] 2. Test parameters:

[1200] hERG concentration: 1×

[1201] Tracer concentration: 1 nM

[1202] Incubation time: 2 hours

[1203] BMG PHERAstar FS FP

[1204] 3. Test methods:

[1205] Follow the instructions in the kit to perform the experiment, as follows:

[1206] Test group: Different concentrations of the test compound were added to microplates containing hERG cell membranes. Tracer, a tracer with high hERG affinity, was added to each well. After incubating the microplates at room temperature for 2 hours, the changes in fluorescence polarization (Excitation: 540nm; Emission: 590nm) values ​​were detected using a multi-functional microplate reader.

[1207] Positive control group: The test compound was replaced with 30 μM positive control compound E4031, and the experimental method was the same as that of the test group.

[1208] Blank control group: hERG buffer was used instead of the test compound, and no hERG cell membrane was added. The experimental method was the same as that of the test group.

[1209] 4. Data Processing:

[1210] The percentage inhibition rate of the compounds of the present invention against hERG at different concentrations was calculated according to the following formula.

[1211] Percentage inhibition rate = (1 - (fluorescence polarization value of the test compound - fluorescence polarization value of the positive control group) / (fluorescence polarization value of the blank control group - fluorescence polarization value of the positive control group)) * 100%

[1212] When the percentage inhibition rate is between 30% and 80%, the half-maximal inhibitory concentration (IC50) of the compound against hERG is estimated according to the following formula. 50 ) or range:

[1213] IC 50 =X×(1-percentage inhibition rate (%)) / percentage inhibition rate (%), where: X is the test concentration of the compound when the inhibition rate is between 30-80%.

[1214] 5. Experimental Results:

[1215] The inhibition of hERG by the compound was determined using the above method, and the results are shown in Table 5 below.

[1216] Table 5. Results of hERG inhibition test

[1217] Compound numbering <![CDATA[IC 50 (μM)]]> 3 >10 5 >10 7 >10 14 >10 17 >10 19 >10 29 >10 31 >10 32 >10 34 >10 35 >10 39 >10 40 >10 41 >10 42 >10 43 >10 44 >10 45 >10 46 >10 47 >10 48 >10 50 >10 53 >10 55 >10

[1218] 56 >10 57 >10 59 >10 62 >10 63 >10 65 >10

[1219] Test results show that the compound of this invention has low affinity for hERG and competes with the affinity tracer Tracer for IC50. 50 All values ​​are >10 μM. This demonstrates that the compounds of this invention, hERG, have a low risk of cardiotoxicity related to ion channels.

[1220] Experimental Example 6: Stability of Compound Metabolism in Liver Microsomes

[1221] 1. Materials and Methods

[1222] 1.1 Main Test Materials

[1223] Testosterone Dr. Ehrenstorfer, Germany

[1224] Mixed human liver microsomes from Xenotech or BioIVT, USA

[1225] BioIVT in liver microsomes from mixed male SD rats, USA

[1226] 1.2 Liver microsomal incubation system

[1227] Mix the positive compound testosterone or the analyte (liver microsomal solution, 50 μl) with PBS (25 μl), pre-incubate (37 °C) for 5 min, then add NADPH (25 μl) to bring the final concentration of the positive compound or analyte to 1 μM, and the final concentration of human and rat liver microsomal protein to 0.5 mg / ml. Incubate the analyte group and the positive compound group for 0 and 15 min respectively. After the corresponding reaction time, add 300 μl of ice-cold acetonitrile containing the internal standard to terminate the reaction, vortex, and store at -80 °C for analysis. All incubated samples were duplicates.

[1228] 1.3 Sample Pretreatment

[1229] Vortex the sample for 1 min, then centrifuge for 10 min (4℃, 4000 rpm). Take 160 μl of the supernatant, add 160 μl of 50% acetonitrile-water, mix well, and then perform LC-MS / MS analysis.

[1230] 2. Data Processing

[1231] Using Excel software, calculate the remaining percentage (%) of the parent drug in the incubation system:

[1232] Original residual rate (%) = 100 × (A) 孵育样品 / A 0h )

[1233] Note: A 孵育样品 : Peak area ratio of the compound to the internal standard after the corresponding incubation time; A 0h : The ratio of the peak area of ​​the unreacted compound to that of the internal standard.

[1234] 3. Results and Conclusions

[1235] The original residual rate of the compound after incubation in human and rat liver microsomes for 15 min was determined according to the above method, as shown in Table 6.

[1236] Table 6. Residual fraction of the compounds of the present invention after incubation in human and rat liver microsomes for 15 min.

[1237]

[1238] in conclusion:

[1239] Most of the compounds of this invention are metabolically stable in human liver microsomes and also relatively stable in rat liver microsomes, showing good overall metabolic stability.

[1240] Test Example 7: Permeability Test of Compound MDCK

[1241] 1. Experimental Materials

[1242]

[1243]

[1244] 2. Experiment Content

[1245] 2.1 Reagent Preparation

[1246] Complete culture medium: Add 10% fetal bovine serum and 1% penicillin / streptomycin to MEM culture medium.

[1247] Digestive fluid: 0.05% trypsin-EDTA

[1248] HBSS buffer: containing Ca 2+ Mg 2+ HBSS

[1249] Osmotic fluid: HBSS containing 10 mM Hepes and 2% BSA, pH 7.4

[1250] Compound stock solution: Weigh a certain amount of the compound to be tested and prepare a 10 mM stock solution with DMSO.

[1251] 2.2 Cell Culture

[1252] MDCK cells in logarithmic growth phase (passage 20) were digested and dispersed evenly with 0.05% trypsin-EDTA, and then divided at a ratio of 3*10 cells / cells. 5 pcs / cm 2 Inoculate into Transwell chambers, 200 μl for side A and 1000 μl for side B. Change the medium once a day after inoculation and incubate for 3–4 days.

[1253] 2.3 Permeability Test

[1254] Gently aspirate the cell culture medium, wash three times with osmotic buffer, add 200 μl of osmotic buffer to side A and 1000 μl of osmotic buffer to side B, and measure the transmembrane resistance (TEER) of the cells using a voltage resistance meter.

[1255] After measuring the TEER value of the cells, gently aspirate the osmotic fluid from both sides. Add the solution according to the following system:

[1256]

[1257] After 120 min, 150 μl of samples from side A and side B were taken and stored at -80℃ for later analysis.

[1258] Subsequently, 10 μg / ml fluorescein was added to side A, and after incubation for 30 min, 100 μl of the solution from side B was aspirated into a 96-well white plate. The fluorescence value was measured using a microplate reader at wavelengths of Ex = 485 nm and Em = 530 nm, and the transmittance (less than 1%) was calculated.

[1259] 2.4 Sample Preparation

[1260] Take 80 μl of fenoterol sample and add 320 μl of acetonitrile solution containing internal standard (10 ng / ml Terfenadine), vortex, and centrifuge for 10 min (4000 rpm, 4℃). The supernatant is diluted 1:1 with 50% acetonitrile-water and then analyzed by LC-MS / MS.

[1261] Add 40 μl of propranolol sample to 360 μl of acetonitrile solution containing internal standard (10 ng / ml Terfenadine), vortex, and centrifuge for 10 min (4000 rpm, 4℃). Perform LC-MS / MS analysis on the supernatant.

[1262] Take 80 μl of digoxin sample and add 320 μl of acetonitrile solution containing internal standard (10 ng / ml Terfenadine), vortex, and centrifuge for 10 min (4000 rpm, 4℃). Take the supernatant for LC-MS / MS analysis.

[1263] Add 40 μl of the analyte to 320 μl of acetonitrile solution containing the internal standard (10 ng / ml Terfenadine), vortex, and centrifuge for 10 min (4000 rpm, 4℃). Perform LC-MS / MS analysis on the supernatant.

[1264] 2.5 Data Processing

[1265] Calculate P using Excel 2013 app Values ​​and P values ​​from side B to side A and from side A to side B. app The ratio was calculated, and the recovery rate of each positive compound and the test compound was determined.

[1266] 2.5.1 Calculation of apparent permeability

[1267] The apparent permeability coefficient (P) of the compound is calculated according to the following formula. app ):

[1268] P app = (dCr / dt)×Vr / (A×C0)

[1269] Where C0 is the initial concentration (μM) of the test drug at the donor end, (dCr / dt)×Vr is the rate at which the test drug appears at the receiver end (μM / s), Vr is the solution volume on the receiver side (ml), and A is the surface area of ​​the polycarbonate membrane (cm²). 2 ).

[1270] 2.5.2 Transporter

[1271] According to P app Calculate the efflux ratio (ER):

[1272] ER=P app (B→A) / P app (A→B)

[1273] Among them, P app (B→A) represents the apparent permeability coefficient of the test drug from the BL (baso-lateral) end to the AP (apical) end, P app(A→B) The apparent permeability coefficient of the test drug from the AP end to the BL end.

[1274] 3. Results and Conclusions

[1275] The permeability values ​​of the compounds were determined using the method described above, as shown in Table 7.

[1276] Table 7. Permeability values ​​of the compounds of this invention

[1277]

[1278] in conclusion:

[1279] The test compounds of this invention are all moderately permeable compounds.

[1280] Experimental Example 8: Pharmacokinetic Study of Compounds in SD Rats

[1281] The compound of the present invention was administered to male SD rats via intravenous injection (IV) and oral gavage (PO) to investigate its pharmacokinetic characteristics.

[1282] The compounds of this invention were administered intravenously and orally, with the dosages shown in Table 8. Blood samples were collected before IV administration (0 h) and at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h after IV administration, and before PO administration (0 h) and at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h after PO administration. 0.3 ml of blood was collected via tail vein and placed in a K2-EDTA anticoagulant tube. The plasma was centrifuged at 4000 rpm for 5 min (4℃) and stored at -80℃ for analysis. After protein precipitation, the plasma samples were analyzed by LC-MS / MS. Pharmacokinetic parameters were calculated using a non-compartmental model with WinNonlin 6.3 software, and the results are shown in Table 8.

[1283] Table 8: Pharmacokinetic parameters of the compounds of the present invention in rats

[1284]

[1285]

[1286] a) A mixture of 10% DMSO, 10% Solutol (polyethylene glycol-15-hydroxystearate), and 80% physiological saline; b) A mixture of 40% Solutol (polyethylene glycol-15-hydroxystearate) and 60% pure water; c) A mixture of 5% DMSO, 5% Solutol (polyethylene glycol-15-hydroxystearate), and 90% physiological saline; d) A mixture of 10% DMSO, 10% Solutol (polyethylene glycol-15-hydroxystearate), and 80% physiological saline (pH=2).

[1287] in conclusion:

[1288] Compound 43 of this invention exhibits high exposure, good bioavailability, and a relatively long half-life in rats after administration via PO, demonstrating excellent overall pharmacokinetic properties. Compounds 17, 41, and 63 also showed good plasma exposure and bioavailability after PO administration. This indicates that the compounds of this invention all possess good oral absorption in rats.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, I in, R 1 The group is selected from C1-C6 alkyl, C3-C6 cycloalkyl and 3-6 oxocyclic alkyl, wherein the C1-C6 alkyl, C3-C6 cycloalkyl or 3-6 oxocyclic alkyl is optionally substituted by one or more groups each independently selected from: halogen, -CN, -OH, -S(O)2 (C1-C6 alkyl), C3-C6 cycloalkyl optionally substituted with halogen or =O, and phenyl optionally substituted with halogen; R 2 Each time it appears, it is independently selected from H and halogen; m is selected from 0, 1, and 2; R 3 Selected from phenyl and 5-6 nitrogen-containing heteroaryl groups, wherein the phenyl or 5-6 nitrogen-containing heteroaryl group is optionally substituted by one or more groups each independently selected from the following groups: halogen, C1-C6 alkyl and C1-C6 haloalkyl; R 4 Each of the following groups, when appearing independently, is selected from H, -OH, halogen, CN, -NH2, -NO2, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, and 3-8 membered heterocyclic groups, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, or 3-8 membered heterocyclic group is optionally replaced by one or more R groups. 9 replace; R 9 Each of these groups is independently selected from -OH, halogen, -CN, -NH2, -NO2, C1-C6 alkoxy, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy, and 3-8 membered heterocyclic groups when it appears. n is selected from 0, 1, and 2; and X is selected from CH and N.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 1 The group is selected from C1-C6 alkyl, C3-C6 cycloalkyl, oxetyl, tetrahydrofuranyl, and tetrahydropyranyl, wherein the C1-C6 alkyl or C3-C6 cycloalkyl is optionally substituted with a group selected from: halogen, -CN, -OH, -S(O)2 (C1-C6 alkyl), C3-C6 cycloalkyl optionally substituted with halogen or =O, and phenyl optionally substituted with halogen.

3. The compound of claim 2 or a pharmaceutically acceptable salt thereof, wherein, R 1 The group is selected from C1-C6 alkyl, C3-C6 cycloalkyl, oxetyl, tetrahydrofuranyl and tetrahydropyranyl, wherein the C1-C6 alkyl or C3-C6 cycloalkyl is optionally substituted with a group selected from the following: -F, -CN, -OH, -S(O)2CH3, C3-C6 cycloalkyl, oxocyclohexyl and fluorophenyl.

4. The compound of claim 3 or a pharmaceutically acceptable salt thereof, wherein, R 1 Selected from , , , , , , , , CF3CH2- , , , , , and , Among them, the wavy line This indicates the connection point between the group and the rest of the molecule.

5. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 2 Each time it appears, it is independently selected from H and F.

6. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, m is 1.

7. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 3 Selected from phenyl and pyridyl, wherein the phenyl or pyridyl group is optionally substituted by one or more groups each independently selected from the following groups: halogen, C1-C6 alkyl and C1-C6 haloalkyl.

8. The compound of claim 7 or a pharmaceutically acceptable salt thereof, wherein, R 3 Selected from phenyl and pyridyl, wherein the phenyl or pyridyl group is optionally substituted by one or more groups, each independently selected from the following: F, Cl, -CF3 and -CH3.

9. The compound of claim 8 or a pharmaceutically acceptable salt thereof, wherein, R 3 Selected from: , , , , , and , Among them, the wavy line This indicates the connection point between the group and the rest of the molecule.

10. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 4 Each group is independently selected from C1-C6 alkyl and C3-C8 cycloalkyl groups in each occurrence, wherein the C1-C6 alkyl group is optionally substituted by one or more groups independently selected from C1-C6 alkoxy and 3-8 membered heterocyclic groups.

11. The compound of claim 10 or a pharmaceutically acceptable salt thereof, wherein, R 4 Each of the components is independently selected from C1-C6 alkyl and C3-C8 cycloalkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with C1-C6 alkoxy groups or 5-6 member oxygen-containing heterocyclic alkyl groups.

12. The compound of claim 11 or a pharmaceutically acceptable salt thereof, wherein, R 4 Each of the components is independently selected from C1-C6 alkyl and C3-C8 cycloalkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with C1-C6 alkoxy or tetrahydropyranyl groups.

13. The compound of claim 12 or a pharmaceutically acceptable salt thereof, wherein, R 4 Each time it appears, it is independently selected from isopropyl... , and , Among them, the wavy line This indicates the connection point between the group and the rest of the molecule.

14. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein, n is 1.

15. A compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of formula I-1: I-1。 16. A compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, wherein said compound is a compound of formula I-2: I-2。 17. A compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of formulas I-3: I-3 in, q is selected from 0, 1, 2, 3, 4 and 5.

18. The compound of claim 17 or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of formula I-3: wherein, q is 1.

19. A compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of formulas I-4: I-4 in, r is selected from 0, 1, 2, 3, and 4.

20. The compound of claim 19 or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of formula I-4: wherein, r is 1.

21. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: 。 22. A pharmaceutical composition comprising a preventive or therapeutically effective amount of the compound of any one of claims 1-21 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

23. A medicine box, comprising: a) the compound of any one of claims 1-21 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 22; and b) Optional packaging and / or instructions.

24. A method for preparing the compound according to any one of claims 1-21, comprising the steps shown in reaction route 1 or reaction route 8: Reaction route 1 Reaction route 8 in, R 1 R 2 R 3 R 4 X, m, and n are as defined in any one of claims 1-21.

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