A PAK4 kinase inhibitor, its preparation method and uses

Abstract

This invention discloses a compound of formula I, or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof. Experimental results show that the compound provided by this invention exhibits high inhibitory activity against PAK4 kinase and PAK I / II selectivity, good liver microsomal stability, and rat PK pharmacokinetic properties, especially with low hERG cardiotoxicity risk, representing a significant improvement over prior art compounds.

Description

Technical Field

[0001] This invention belongs to the field of drug synthesis technology, specifically relating to a PAK4 kinase inhibitor, its preparation method, and its uses. Background Technology

[0002] P21-activated protein kinases (PAKs), a class of conserved serine / threonine protein kinases, are effector proteins of the small GTPases CDC42 and Rac1 in the Rho family, mediating the transduction of their downstream signaling pathways. Based on their sequence homology and activation modes, they can be divided into two main classes: class I PAKs (PAK1, 2, 3) and class II PAKs (PAK4, 5, 6). As important downstream molecules of the Pho family GTPases Rac and Cdc42, PAKs play crucial roles in cell proliferation, cytoskeleton remodeling, and cell motility. Studies have shown that PAK members, especially their representative members PAK1 and PAK4, exhibit gene amplification, gene mutation, upregulated expression levels, and activity in various tumor cells and tissues, which are closely related to tumorigenesis and development. Inhibiting the abnormal activity of PAKs within tumor cells holds promise for suppressing excessive tumor cell proliferation, invasion, metastasis, and angiogenesis, and promoting tumor cell apoptosis.

[0003] In light of this, research on PAK inhibitors has received widespread attention from medicinal chemists over the past decade. Studies by Wang C et al. have shown that PAK4 expression levels in lung cancer, colon cancer, prostate cancer, pancreatic cancer, and breast cancer cells are significantly higher than in normal cells, exerting a crucial influence on tumor occurrence, development, invasion, and migration. Therefore, the development of PAK4 inhibitors is one of the effective strategies for treating various cancers.

[0004] Recent studies have revealed a potential correlation between inhibition of class I PAKs and safety risks such as acute cardiotoxicity and hERG side effects, suggesting that the development of PAK inhibitors should avoid inhibition of class I PAKs, especially PAK1. Therefore, the discovery of highly selective class II PAK inhibitors will become the mainstream of future research.

[0005] PAK4 is a promising drug development target, and the development of its inhibitors offers new avenues for treating related cancers. To date, the number of PAK4 inhibitors is limited, and most exhibit unsatisfactory activity. Furthermore, poor pharmacokinetic properties and druggability issues are common problems for reported molecules. Currently, small molecule inhibitors reported to be in clinical trials include ATG-019 (KPT-9274), jointly developed by Antengene and Karyopharm Therapeutics, and PF-3758309 developed by Pfizer. PF-3758309, a class I PAKs inhibitor with a pyrrolopyrazole structure reported by Pfizer in 2009, was the first PAKs inhibitor to enter clinical trials. Its PAK4 IC50 is 19 nm, but this compound exhibits stronger inhibitory activity against PAK1, reaching 14 nm, posing significant safety risks. Additionally, its poor oral bioavailability (approximately 1%) and severe gastrointestinal adverse reactions forced the termination of its Phase I clinical trial. ATG-019 is a first-in-class oral dual-target inhibitor of p21-activated kinase 4 (PAK4) and nicotinic acid transphosphoribosylase (NAMPT), with an unknown mechanism of action. Multiple Phase I clinical trials are currently underway in areas including non-Hodgkin's lymphoma, colorectal cancer, lung cancer, and melanoma. Summary of the Invention

[0006] Our company previously developed compounds that can act as PAK4 kinase inhibitors, disclosed in patent WO2022033420A1. These compounds all exhibit good PAK4 activity and PAK I / II selectivity, representing a significant improvement in pharmacokinetic properties compared to previously reported PAK4 kinase inhibitors. However, in subsequent research, the inventors discovered that these compounds possess strong hERG cardiac depressant activity, leading to an increased risk of cardiotoxicity. Therefore, based on previous research, this invention optimizes the compound structures and screens a series of compounds with higher inhibitory activity and PAK I / II selectivity for PAK4 kinase. These compounds not only possess excellent liver microsomal stability and rat PK pharmacokinetic properties, but also exhibit significantly reduced hERG cardiac depressant activity, thus resolving the cardiotoxicity risk issue.

[0007] The purpose of this invention is to provide a compound of general formula I and a method for preparing the same, wherein the compound is a PAK4 kinase inhibitor; another purpose of this invention is to provide a use of the compound.

[0008] The technical solution of this invention includes the following:

[0009] In a first aspect, the present invention provides a compound of formula I, or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof:

[0010]

[0011] Wherein, A1, A2, A3, and A4 are independently selected from C or N. When any one of A1, A2, A3, and A4 is N, R2, R3, R4, and R5 connected to N do not exist.

[0012] In a preferred embodiment of the present invention, A1, A2, A3, and A4 are selected from C.

[0013] R2, R3, R4, and R5 are independently selected from -H, halogen, -OH, -CN, -NH2, -NO2, -SH, C1-10 straight / branched alkyl, C3-10 cycloalkyl, heterocyclic alkyl, alkynyl, alkenyl, aromatic, heterocyclic aromatic, amide, ester, sulfonyl, and phosphoryl groups, wherein the H on the above groups can be substituted by the following groups: halogen, -OH, -CN, -NH2, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl or heterocyclic alkyl, substituted or unsubstituted aromatic or heterocyclic aromatic.

[0014] The substituents are selected from halogens, -OH, -CN, -NH2, -NO2, -SH, carboxyl, hydroxyamino, alkyl, cycloalkyl, heterocycloalkyl, aromatic, heterocyclic aromatic, ester, acyl, carbonyl, amide, sulfonyl, and phosphoryl.

[0015] The cycloalkyl or heterocycloalkyl groups described in this invention include monocyclic, bridged, spirocyclic, or fused-ring cycloalkyl or heterocycloalkyl groups. Further, the cycloalkyl or heterocycloalkyl groups include both saturated and unsaturated forms.

[0016] B1 and B2 are independently selected from C or N. In a preferred embodiment of the present invention, B1 and B2 are N.

[0017] L is selected from -O-, -S-, -NH- or alkylene; preferably, L is -NH-.

[0018] R1 is selected from substituted or unsubstituted five- or six-membered aromatic or heterocyclic aromatic groups, or substituted or unsubstituted heterocyclic alkyl groups containing at least one N and / or O atom.

[0019] Preferably, R1 is selected from substituted or unsubstituted phenyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, pyridinyl, pyrimidinyl, pyridazinyl, furanyl, thiopheneyl, pyrroleyl, and five- or six-membered heterocyclic alkyl containing at least one N or O atom.

[0020] In a specific embodiment of the present invention, R1 has any of the following structures:

[0021]

[0022] The H atom on any one or more C atoms in the above structure may be substituted by the following groups: -F, -Cl, -Br, -OH, -CN, -NH2, substituted or unsubstituted amide group, substituted or unsubstituted C1-3 alkyl or alkoxy group, C3-6 cycloalkyl group.

[0023] Preferably, the substituents are independently selected from -F, -Cl, -Br, -OH, -CN, -NH2, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxymethyl, and hydroxyethyl.

[0024] R6 is selected from -H, -NH2, -OH, halogen, amide group, sulfonyl group, sulfonic acid group, C1-6 alkyl or alkoxy group, C3-6 cycloalkyl group, C3-6 heterocyclic alkyl group, amino group, C6-12 aromatic group, C5-12 heterocyclic aromatic group; the H on the above C1-6 alkyl or alkoxy group, C3-6 cycloalkyl group, C3-6 heterocyclic alkyl group, amino group, C6-12 aromatic group, C5-12 heterocyclic aromatic group may optionally be substituted by one or more halogen, -OH, -CN, -NH2, -NO2, -SH, sulfonic acid group.

[0025] The ring Ar and the ring Rg are fused together, and the fused bond is any bond on the ring Ar;

[0026] The cyclic Ar is selected from aromatic five-membered heterocyclic groups, aromatic six-membered heterocyclic groups, or phenyl groups. The aromatic five-membered heterocyclic group is selected from: imidazolyl, thiazolyl, oxazolyl, pyrrolel, pyrazolyl, furanyl, or thiophenel. The aromatic six-membered heterocyclic group is selected from: pyridinyl, pyridazinyl, pyrimidinyl, or pyrazinyl. Optionally, the H on the aromatic five-membered heterocyclic group, aromatic six-membered heterocyclic group, or phenyl group may be substituted with the following groups: halogen, -OH, -CN, -NH2, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aromatic group or heterocyclic aromatic group. The substituted group is selected from halogen, -OH, -CN, -NH2, -NO2, -SH, carboxyl, hydroxyamino, alkyl, cycloalkyl, heterocycloalkyl, aromatic group, heterocyclic aromatic group, ester group, acyl group, carbonyl group, amide group, sulfonyl group, or phosphoryl group.

[0027] The ring Rg is selected from C3-8 saturated / unsaturated cycloalkyl groups or C3-8 saturated / unsaturated heterocycloalkyl groups containing at least one O, N, or S. Optionally, the H on the C3-8 saturated / unsaturated cycloalkyl group or the C3-8 saturated / unsaturated heterocycloalkyl group containing at least one O, N, or S may be substituted with the following groups: halogen, -OH, -CN, -NH2, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl or heterocycloalkyl, substituted or unsubstituted aromatic or heterocyclic aromatic groups. The substituted group is selected from halogen, -OH, -CN, -NH2, -NO2, -SH, carboxyl, hydroxyamino, alkyl, cycloalkyl, heterocycloalkyl, aromatic, heterocyclic aromatic, ester, acyl, carbonyl, amide, sulfonyl, and phosphoryl.

[0028] Furthermore, the ring Rg has at least one R7, which is independently selected from -H, deuterium, tritium, halogen, -OH, -CN, -NH2, -NO2, -SH, carboxyl, hydroxyamino, alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, aryl, heteroaryl, ester, acyl, carbonyl, amide, sulfonyl, and phosphoryl.

[0029] Preferably, the compound has the structure shown in Formula II, or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof:

[0030]

[0031] X is selected from substituted or unsubstituted C1-6 straight-chain / branched alkyl groups, substituted or unsubstituted C3-10 cycloalkyl or heterocycloalkyl groups, substituted or unsubstituted C5-6 aromatic or heterocyclic aromatic groups, alkynyl, alkenyl, amide, and ester groups. The substituent group is selected from halogen, -OH, -CN, -NH2, -NO2, -SH, carboxyl, hydroxyamino, alkyl, alkoxy, hydroxyalkyl, cycloalkyl, heterocyclic alkyl, aromatic, heterocyclic aromatic, ester, acyl, carbonyl, amide, sulfonyl, and phosphoryl groups.

[0032] The cycloalkyl or heterocycloalkyl groups described in this invention include monocyclic, bridged, spirocyclic, or fused-ring cycloalkyl or heterocycloalkyl groups. Further, the cycloalkyl or heterocycloalkyl groups include both saturated and unsaturated forms.

[0033] The X group has at least one R8, wherein the R8 is independently selected from -H, halogen, -OH, -CN, -NH2, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl or heterocycloalkyl, wherein the substituted group is selected from halogen, -OH, -CN, -NH2, -NO2, -SH, -CF3, -CHF2, -CH2F, carboxyl, hydroxyamino, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl, phenyl, phenoxy, pyridyl, amide, sulfonyl, phosphoryl.

[0034] More preferably, the substituted groups are independently selected from -F, -Cl, -Br, -OH, -CN, -NH2, -CF3, hydroxymethyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0035] R9 is selected from substituted or unsubstituted phenyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and imidazolyl groups, wherein the substituent group is independently selected from -F, -Cl, -Br, -OH, -CN, -NH2, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and hydroxymethyl.

[0036] R 10 The H on the C1-6 alkyl or alkoxy group, C3-6 cycloalkyl group, or C3-6 heterocyclic alkyl group may optionally be replaced by one or more halogens, -OH, -CN, -NH2, -NO2, or -SH.

[0037] Ring Rg is selected from C3-8 saturated / unsaturated heterocyclic alkyl groups containing at least one O, N, or S, and ring Rg has at least one R. 11 R 11 Independently selected from -H, deuterium, tritium, -F, -Cl, -Br, -OH, -CN, -NH2, -NO2, -SH, hydroxyamino, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl, phenyl, phenoxy, sulfonyl, phosphoryl.

[0038] In a preferred embodiment of the invention, the compound has the structure shown in Formula III, or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof:

[0039]

[0040] R 12 Selected from -H, -F, -Cl, -Br, -OH, -CN, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxymethyl.

[0041] R 13 R 14 Independently selected from -H, deuterium, tritium, -F, -Cl, -Br, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl.

[0042] R 15 The group is selected from -H, -F, -Br, -OH, -CN, -NH2, substituted or unsubstituted C1-10 straight / branched alkyl, substituted or unsubstituted C1-10 straight / branched alkoxy, substituted or unsubstituted cycloalkyl or heterocycloalkyl, wherein the H on the above groups may be substituted by the following groups: -F, -Cl, -Br, -OH, -CN, -NH2, -CF3, hydroxymethyl, methyl, ethyl, propyl, isopropyl, phenyl, methoxy, ethoxy, propoxy, isopropoxy, phenoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0043] The cycloalkyl or heterocycloalkyl group includes monocyclic, bridged, spirocyclic, or fused-ring cycloalkyl or heterocycloalkyl groups. Further, the cycloalkyl or heterocycloalkyl group includes both saturated and unsaturated forms.

[0044] In specific embodiments of the present invention, the substituted or unsubstituted cycloalkyl or heterocycloalkyl groups include, but are not limited to, those comprising: Indicates the position where it can be attached to an alkynyl group.

[0045] In a preferred embodiment of the invention, the compound has the structure shown in Formula IV, or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof:

[0046]

[0047] R 16 R 17 Independently selected from -H, -F, -Cl, -Br, -OH, -CN, -NH2, -SH, hydroxyamino, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl.

[0048] R 18Selected from -H, -F, -Cl, -Br, -OH, -CN, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxymethyl.

[0049] R 19 R 20 Independently selected from -H, deuterium, tritium, -F, -Cl, -Br, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl.

[0050] In a preferred embodiment of the invention, the compound has the structure shown in Formula V, or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof:

[0051]

[0052] R 21 Selected from -H, -F, -Cl, -Br, -OH, -CN, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxymethyl.

[0053] R 22 R 23 Independently selected from -H, deuterium, tritium, -F, -Cl, -Br, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl.

[0054] Y is selected from substituted or unsubstituted C1-4 straight-chain / branched alkyl groups, substituted or unsubstituted C3-8 cycloalkyl or heterocyclic alkyl groups, substituted or unsubstituted C5-6 aromatic or heterocyclic aromatic groups, amide groups, and ester groups. The substituent group is selected from -OH, -CN, -NH2, -NO2, -SH, carboxyl, hydroxyamino, alkyl, alkoxy, hydroxyalkyl, substituted or unsubstituted C3-8 cycloalkyl or heterocyclic alkyl groups, C5-6 aromatic or heterocyclic aromatic groups, ester groups, acyl groups, carbonyl groups, amide groups, sulfonyl groups, and phosphoryl groups.

[0055] The R 24 Selected from -H, deuterium, tritium, -F, -Cl, -Br, -OH, -CN, -NH2, -NO2, -SH, hydroxyamino, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, phenyl, phenoxy, substituted or unsubstituted C3-8 saturated or unsaturated cycloalkyl or heterocycloalkyl.

[0056] The substituted or unsubstituted C3-8 cycloalkyl or heterocycloalkyl groups include, but are not limited to, those mentioned above. This indicates the position where Y can be attached. The H in the above structure can be substituted by the following groups: -OH, -CN, -NH2, -NO2, -SH, hydroxyamino, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, phenyl, phenoxy, sulfonyl, phosphoryl.

[0057] In the most preferred embodiment of the present invention, the compound has the following structure:

[0058]

[0059]

[0060] In a second aspect, the present invention provides a pharmaceutical composition comprising a compound of any one of general formulas I-V or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof.

[0061] Preferably, the pharmaceutical composition further includes pharmaceutically acceptable excipients, including but not limited to: carriers, diluents, binders, lubricants, and wetting agents.

[0062] Preferably, the pharmaceutical composition comprises a therapeutically effective amount of any one of the compounds of formulas I-V. In some embodiments, the pharmaceutical composition may be used alone or in combination with other formulations.

[0063] The pharmaceutical composition is suitable for gastrointestinal or non-gastrointestinal administration, such as via intravenous, intramuscular, intradermal, and subcutaneous routes. Preferably, the pharmaceutical composition further includes an antioxidant, a buffer, an antibacterial agent, and a solute that makes the formulation isotonic with the subject's blood, as well as an aqueous and non-aqueous sterile suspending agent, which may contain suspending agents, solubilizers, thickeners, stabilizers, and preservatives.

[0064] The compounds of any one of general formulas I-V provided by this invention can be formulated into pharmaceutical preparations in the following forms: injections, syrups, elixirs, suspensions, powders, granules, tablets, capsules, lozenges, creams, ointments, lotions, gels, emulsions, etc.

[0065] Thirdly, the present invention provides the use of a compound of any one of general formulas I-V or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof, or a pharmaceutical composition comprising a compound of any one of general formulas I-V or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate thereof, in the preparation of a medicament for treating diseases related to the expression or activity of PAK4 kinase.

[0066] The diseases associated with the expression or activity of PAK4 kinase include cancer, neurodegenerative diseases, or immune system diseases.

[0067] The cancers mentioned include breast cancer, mantle cell lymphoma, ovarian cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, hepatocellular carcinoma, glioma, endometrial cancer, melanoma, kidney cancer, bladder cancer, biliary tract cancer, pancreatic cancer, lymphoma, pilocytic carcinoma, nasopharyngeal carcinoma, pharyngeal cancer, colorectal cancer, rectal cancer, brain and central nervous system cancers, cervical cancer, prostate cancer, testicular cancer, and urogenital tract cancers. Lung cancer, non-small cell lung cancer, small cell carcinoma, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, adenocarcinoma, thyroid cancer, follicular carcinoma, Hodgkin's leukemia, bronchial cancer, thyroid cancer, endometrial cancer, cervical cancer, multiple myeloma, acute myeloid leukemia, chronic myeloid leukemia, lymphocytic leukemia, chronic lymphocytic leukemia, myeloid leukemia, non-Hodgkin's lymphoma, primary macroglobulinemia, rhabdomyosarcoma.

[0068] Preferably, the compound of any one of general formulas I-V or its pharmaceutically acceptable salt, stereoisomer, ester, prodrug, or solvate may be used alone or in combination with other types of pharmaceutical preparations and / or treatment methods.

[0069] Other types of pharmaceutical preparations and / or treatments include, but are not limited to: immunosuppressants, targeted anti-tumor drugs, glucocorticoids, nonsteroidal anti-inflammatory drugs, anti-tumor vaccines, adoptive cellular immunotherapy, chemotherapy, or radiotherapy.

[0070] Fourthly, the present invention provides a compound of the general formula VI.

[0071]

[0072] Among them, R 25 R 26 Independently selected from -H, deuterium, tritium, -F, -Cl, -Br, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl.

[0073] Fifthly, the present invention provides a compound of formula VII.

[0074]

[0075] Among them, R 27 Selected from -H, -F, -Cl, -Br, -OH, -CN, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hydroxymethyl.

[0076] R 28 R 29Independently selected from -H, deuterium, tritium, -F, -Cl, -Br, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl.

[0077] Sixthly, the present invention provides the use of compounds of formula VI and / or formula VII in the preparation of any of the following compounds, or pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, or solvates thereof:

[0078] (a) The compound described in Formula I;

[0079] (b) The compound described in Formula II;

[0080] (c) The compound described in Formula III;

[0081] (d) The compound described in Formula IV;

[0082] (e) The compound described in formula V.

[0083] The technical solution provided by the present invention has the following technical advantages: (1) Compared with the prior art, the compounds of general formula I-V or their pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, and solvates provided by the present invention have high inhibitory activity and PAK I / II selectivity as PAK4 inhibitors, while their liver microsomal stability and rat PK are also improved; (2) Furthermore, the compounds of general formula I or their pharmaceutically acceptable salts, stereoisomers, esters, prodrugs, and solvates provided by the present invention have greatly reduced hERG cardiac inhibitory activity as PAK4 inhibitors, thus solving the risk of drug cardiotoxicity. Detailed Implementation

[0084] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0085] The abbreviations appearing in the following examples have the following meanings:

[0086] Et3N: Triethylamine;

[0087] EA: Ethyl acetate;

[0088] THF: Tetrahydrofuran;

[0089] MeOH: Methanol;

[0090] DIEA: N,N-diisopropylethylamine;

[0091] n-BuLi: n-Butyllithium;

[0092] M: Molar concentration unit mol / L, for example 1M means 1 mol / L;

[0093] N: Equivalent concentration, for example, 1N HCl refers to hydrochloric acid with a concentration of 1 mol / L;

[0094] HATU: O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate;

[0095] HBTU: Benzotriazole-N,N,N',N'-Tetramethylurea hexafluorophosphate;

[0096] m-CPBA: m-chloroperoxybenzoic acid;

[0097] NMO: N-methylmorpholine oxide;

[0098] DPPA: Diphenyl azidophosphate;

[0099] DMF: N,N-dimethylformamide;

[0100] TBDPSCl: tert-butyldiphenylchlorosilane;

[0101] DIBAL-H: Diisobutylaluminum hydride;

[0102] TLC: Thin-layer chromatography;

[0103] PE: Petroleum ether (boiling point 60-90℃);

[0104] DCM: Dichloromethane;

[0105] H2O: Distilled water;

[0106] DMSO: Dimethyl sulfoxide;

[0107] Pd(PPh3)2Cl2: Palladium dichloride of bis(triphenylphosphine);

[0108] Pd(dppf)Cl2.DCM: [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane complex;

[0109] HCl / 1,4-dioxane: hydrochloric acid / 1,4-dioxane solution;

[0110] TBAF.3H2O: Tetrabutylammonium fluoride trihydrate;

[0111] 1,4-dioxane: 1,4-dioxane.

[0112] Preparation of intermediates

[0113] Intermediate 1 was prepared using the following synthetic route:

[0114]

[0115] The preparation method of compound 4-(benzo[d][1,3]dioxo-5-yl)piperidin-4-amine dihydrochloride is described in detail below:

[0116]

[0117] Step 1: Preparation of tert-butyl 4-(benzo[d][1,3]dioxo-5-yl)-4-cyanopiperidin-1-carboxylic acid

[0118] Weigh 10.00 g (62.03 mmol) of 2-(benzo[d][1,3]dioxol-5-yl)acetonitrile and 16.52 g (68.23 mmol) of bis(2-chloroethyl)carbamate tert-butyl ester into a reaction flask, add 120 mL of DMF, purge twice with nitrogen, and cool to approximately 0 °C. Add NaH (9.93 g, 248.12 mmol, 60% in oil) in portions, maintaining the system temperature between 0 and 10 °C. After the addition is complete, raise the temperature to 60 °C and react for 2 hours. Turn off the heating and allow the mixture to cool naturally to room temperature with stirring, and react for 14 hours. TLC plate analysis was performed on the sample. After the reaction of the raw materials was completed, the reaction solution was slowly added to a saturated NH4Cl aqueous solution (600 mL) for quenching. Then, EA (200 mL * 2) was added, and the mixture was extracted twice. The organic phases were combined and washed three times with a saturated NaCl aqueous solution (100 mL * 3). The mixture was dried over anhydrous sodium sulfate and concentrated to obtain a brown solid (crude product), which was directly used in the next reaction.

[0119] EM (calculated value): 330.2; MS (ESI) m / z (M+H) + 331.3

[0120] Step 2: Preparation of tert-butyl 4-(benzo[d][1,3]dioxo-5-yl)-4-carbamoylpiperidine-1-carboxylic acid

[0121] Compound 4-(benzo[d][1,3]dioxo-5-yl)-4-cyanopiperidin-1-carboxylic acid tert-butyl ester (crude product) and NaOH (24.81 g, 620.30 mmol) were placed in a reaction flask, and DMSO (140 mL) was added. The mixture was heated to 60 °C, and H2O2 (61 mL) was slowly added dropwise. After the addition was complete, a sample was taken to monitor the reaction. After the reaction was complete, the mixture was cooled to room temperature. The reaction solution was slowly poured into water (1120 mL), producing a large amount of pale yellow solid. After stirring for 20 minutes, the mixture was filtered, and the filter cake was washed three times with water. The filter cake was dried to obtain a pale yellow solid (crude product), which was directly used in the next reaction step.

[0122] EM (calculated value): 348.2; MS (ESI) m / z (M+H) + 349.3

[0123] Step 3: Preparation of tert-butyl 4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidine-1-carboxylic acid

[0124] Weigh 4-(benzo[d][1,3]dioxo-5-yl)-4-carbamoylpiperidin-1-carboxylic acid tert-butyl ester (crude product) into a reaction flask and add acetonitrile (250 mL). While stirring, add an aqueous solution of KOH (13.89 g, 248.12 mmol) (KOH was pre-dispersed in 125 mL of water) in an open reaction flask. Slowly add dibromohydantoin (10.64 g, 37.22 mmol) at room temperature. After the addition is complete, the system gradually dissolves and becomes lighter in color. React at room temperature for 2 hours. After the reaction is complete as monitored by TLC, stop stirring, separate the liquids, collect the organic phase, and concentrate off most of the solvent. Extract the aqueous phase twice using DCM / MeOH = 10 / 1 (100 mL * 2). Combine the concentrated residue with the extract, wash once with saturated NaCl aqueous solution, dry with anhydrous sodium sulfate, and concentrate to dryness to obtain a black oily substance (crude product), which is directly used in the next reaction step.

[0125] EM (calculated value): 320.2; MS (ESI) m / z (M+H) + 321.3

[0126] Step 4: Preparation of 4-(benzo[d][1,3]dioxo-5-yl)piperidine-4-amine dihydrochloride

[0127] Weigh 4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidine-1-carboxylic acid tert-butyl ester (crude product) into a reaction flask, and add HCl / 1,4-dioxane (127 mL, 4 M). Stir at room temperature for about 10 minutes until a solid precipitates. Continue stirring for 1 hour until the reaction is complete. Filter the reaction solution directly, and wash the filter cake with a small amount of 1,4-dioxane to obtain a grayish-brown solid compound (15.70 g, overall yield of the four steps: 86.4%).

[0128] EM (calculated value): 220.1; MS (ESI) m / z (M+H) + :221.2

[0129] Intermediate 2

[0130] Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine

[0131]

[0132] Step 1: Preparation of 2-chloro-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine

[0133] Compound 2,4-dichloro-6-iodoquinazoline (5.00 g, 15.39 mmol) was dissolved in DMF (30 mL). 5-cyclopropyl-4-fluoro-1H-pyrazole-3-amine (2.20 g, 15.44 mmol) and DIEA (7.90 g, 61.56 mmol) were added sequentially at room temperature. The mixture was heated to 65 °C and stirred for 2 hours. After the reactants had reacted completely, the reaction solution was cooled to room temperature. The reaction solution was slowly poured into water (300 mL), and extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution, then dried over anhydrous sodium sulfate. The solution was concentrated to dryness, and the residue was purified by column chromatography (DCM / MeOH = 50 / 1) to give the target compound (6.02 g, yield 91.2%) as a yellow solid.

[0134] EM (calculated value): 429.0; MS (ESI) m / z (M+H) + 430.1

[0135] Step 2: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine

[0136] 2-Chloro-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (6.02 g, 14.01 mmol) was dissolved in DMF (30 mL). 4-(benzo[d][1,3]dioxo-5-yl)piperidine-4-amine dihydrochloride (4.11 g, 14.01 mmol), KI (4.65 g, 28.02 mmol), and DIEA (7.23 g, 56.04 mmol) were added sequentially at room temperature. The mixture was heated to 120 °C and stirred for 2 hours. After the reactants had reacted completely, the reaction solution was cooled to room temperature. The reaction solution was slowly poured into water (300 mL), and extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution, then dried over anhydrous sodium sulfate. The residue was concentrated to dryness and purified by column chromatography (DCM / MeOH = 50 / 1) to obtain the target compound (5.42 g, yield 63.1%) as a yellow solid.

[0137] EM (calculated value): 613.1; MS (ESI) m / z (M+H) + : 614.1

[0138] Preparation of compounds in the examples

[0139] Example 1

[0140] Preparation of 4-((2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ethynyl)tetrahydro-2H-pyran-4-ol

[0141]

[0142] Compounds 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (50 mg, 0.08 mmol) and 4-ethyltetrahydro-2H-pyran-4-ol (30 mg, 0.24 mmol) were added to THF (2 mL), followed by CuI (4 mg, 0.02 mmol), Et3N (24 mg, 0.24 mmol), and Pd(PPh3)2Cl2 (15 mg, 0.02 mmol). The mixture was stirred at 50 °C for 4 hours under nitrogen protection. After the reaction was complete, water (20 mL) was added to the reaction solution, and the mixture was extracted three times with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 25 / 1) to obtain the target compound (13 mg, yield 26.6%) as a white solid.

[0143] EM (calculated value): 611.3; MS (ESI) m / z (M+H) + : 612.4

[0144] 1H NMR(400MHz,DMSO-d6)δ0.75-0.79(2H,m),0.91-0.98(2H,m),1.53-1.56(2H,m),1.69-1.81(4H, m),1.83-1.90(3H,m),1.92-2.11(2H,brs),3.42-3.45(2H,m),3.59-3.63(2H,m),3.75-3.82(2H, m),4.24-4.26(2H,m),5.75(1H,s),5.95(2H,s),6.80(1H,d,J=8.0Hz),6.93(1H,d,J=8.0Hz),7. 12(1H,s),7.27(1H,d,J=8.0Hz),7.52(1H,d,J=12.0Hz),8.39(1H,s),9.85(1H,s),12.39(1H,s).

[0145] The compounds shown in Table 1 below were synthesized according to the method described in Example 1:

[0146] Table 1

[0147]

[0148]

[0149]

[0150]

[0151]

[0152] Example 10

[0153] Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-ethynylquinazoline-4-amine

[0154]

[0155] Step 1: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-((trimethylsilyl)ethynyl)quinazolin-4-amine

[0156] Compounds 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (90 mg, 0.15 mmol) and trimethylsilylacetylene (60 mg, 0.60 mmol) were added to DMF (2 mL), followed by CuI (4 mg, 0.02 mmol), Et3N (45 mg, 0.45 mmol), and Pd(PPh3)2Cl2 (22 mg, 0.03 mmol). The mixture was stirred at 45 °C for 4 hours under nitrogen protection. After the reaction was complete, water (20 mL) was added to the reaction solution, and the mixture was extracted three times with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 40 / 1) to obtain the target compound (65 mg, yield 74.3%) as a yellow solid.

[0157] EM (calculated value): 583.3; MS (ESI) m / z (M+H) + 584.4

[0158] Step 2: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-ethynylquinazoline-4-amine

[0159] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-((trimethylsilyl)ethynyl)quinazolin-4-amine (65 mg, 0.11 mmol) was dissolved in methanol (5 mL), and potassium carbonate (30 mg, 0.22 mmol) was added. The mixture was stirred at room temperature for 4 hours. After the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated to dryness. The crude product was purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (15 mg, yield 26.7%) as a white solid.

[0160] EM (calculated value): 511.2; MS (ESI) m / z (M+H) + 512.3

[0161] 1H NMR(400MHz,DMSO-d6)δ0.76-0.79(2H,m),0.93-0.95(2H,m),1.52-1.55(2H,m),1.74-1. 77(2H,m),1.84-1.86(1H,m),1.87-1.90(2H,brs),3.39-3.42(2H,m),4.13(1H,s),4.25- 4.35(2H,m),5.95(2H,s),6.79(1H,d,J=7.6Hz),6.91(1H,d,J=8.4Hz),7.12(1H,s),7.26 (1H,d,J=8.4Hz),7.55(1H,dd,J=1.6Hz,8.4Hz),8.41(1H,s),9.83(1H,s),12.31(1H,s).

[0162] Example 11

[0163] Preparation of 4-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)-1,1,1-trifluorobutyl-3-yn-2-ol)

[0164]

[0165] Step 1: Preparation of 1,1,1-trifluorobutyl-3-yn-2-ol

[0166] Trimethylsilylacetylene (5.19 g, 52.79 mmol) was added to anhydrous THF (30 mL), cooled to -60 °C, and n-BuLi (31.67 mL, 63.35 mmol, 2 M in n-hexane) was slowly added. After stirring for 1 hour, ethyl trifluoroacetate (5.00 g, 35.19 mmol) was slowly added dropwise, and the temperature was maintained while stirring continued for 5 hours. After the reaction was complete, the reaction solution was quenched with saturated NH4Cl aqueous solution, and then water (50 mL) and EA (50 mL) were added and stirred. The mixture was separated, and the aqueous phase was extracted once with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was dissolved in methanol (30 mL), cooled to 0 °C in an ice-water bath, and sodium borohydride (4.00 g, 105.57 mmol) was added in batches, and the mixture was stirred overnight at room temperature. After the reaction was complete, water (50 mL) was slowly added to the reaction solution, and the mixture was extracted twice with ethyl acetate. The organic phases were combined, washed once with saturated NaCl aqueous solution, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the target compound (3.90 g, yield 89.4%), which was a colorless oil.

[0167] EM (calculated value): 124.0; MS (ESI) m / z (M+H) + : 125.0

[0168] Step 2: Preparation of 4-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)-1,1,1-trifluorobutyl-3-yn-2-ol)

[0169] Compounds 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (100 mg, 0.16 mmol) and 1,1,1-trifluorobutyl-3-yn-2-ol (40 mg, 0.32 mmol) were added to DMF (5 mL), followed by CuI (4 mg, 0.02 mmol), Et3N (48 mg, 0.48 mmol), and Pd(PPh3)2Cl2 (22 mg, 0.03 mmol). The mixture was stirred at 45 °C for 4 hours under nitrogen protection. After the reaction was complete, water (50 mL) was added to the reaction solution, and the mixture was extracted three times with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH / NH3.H2O=20 / 1 / 0.05) to obtain the target compound (46 mg, yield 47.2%) as a white solid.

[0170] EM (calculated value): 609.2; MS (ESI) m / z (M+H) + : 610.3

[0171] 1 H NMR(400MHz,DMSO-d6)δ0.76-0.78(2H,m),0.93-0.95(2H,m),1.57-1.63(2H,m) ,1.84-1.87(3H,m),3.45-3.48(2H,m),4.20-4.29(2H,m),5.23-5.28(1H,m),5. 96(2H,s),6.82(1H,d,J=8.0Hz),6.93(1H,d,J=2.0Hz),7.13-7.15(2H,m),7.29 (1H,d,J=8.8Hz),7.56(1H,d,J=8.8Hz),8.47(1H,s),9.92(1H,s),12.32(1H,s).

[0172] Example 12

[0173] Preparation of 4-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)but-3-yne-1,2-diol

[0174]

[0175] Step 1: Preparation of 1-((tert-butyldimethylsilyl)oxy)but-3-yn-2-ol

[0176] Compound 2-((tert-butyldimethylsilyl)oxy)acetaldehyde (200 mg, 1.15 mmol) was added to anhydrous THF (10 mL), and the mixture was cooled to approximately 0 °C using an ice-water bath. Then, acetylenyl magnesium bromide (2.76 mL, 1.38 mmol, 0.5 M in THF) was slowly added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, water (50 mL) was slowly added to the reaction solution, and the mixture was extracted twice with ethyl acetate. The combined organic phases were washed once with a saturated NaCl aqueous solution, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the target compound (189 mg, yield 82.2%) as a yellow oil.

[0177] EM (calculated value): 200.1; MS (ESI) m / z (M+H) + :201.1

[0178] Step 2: Preparation of but-3-yne-1,2-diol

[0179] Compound 1-((tert-butyldimethylsilyl)oxy)but-3-yn-2-ol (189 mg, 0.94 mmol) was added to 2M HCl / MeOH (5 mL), and the mixture was heated to 45 °C and stirred for 5 hours. After the reaction was complete, the reaction solution was concentrated to dryness to obtain the target compound (156 mg, crude product), which was a black oily substance.

[0180] Step 3: Preparation of 4-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)but-3-yne-1,2-diol)

[0181] Compounds 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (70 mg, 0.11 mmol) and but-3-yn-1,2-diol (156 mg, crude) were added to DMF (5 mL). CuI (4 mg, 0.02 mmol), Et3N (33 mg, 0.33 mmol), and Pd(PPh3)2Cl2 (14 mg, 0.02 mmol) were then added. The mixture was stirred at 45 °C for 3 hours under nitrogen protection. After the reaction was complete, water (50 mL) was added to the reaction solution, and the mixture was extracted three times with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (13 mg, yield 20.7%) as a pale yellow solid.

[0182] EM (calculated value): 571.2; MS (ESI) m / z (M+H) + 572.3

[0183] 1 H NMR(400MHz,DMSO-d6)δ0.78-0.82(2H,m),0.97-1.01(2H,m),1.56-1.59(2H,m),1.76-1.78(2H,m ),1.85-1.90(1H,m),3.41-3.45(2H,m),3.62-3.68(2H,m),4.36-4.40(2H,m),4.49-4.55(1H,m),4 .71-4.75(1H,m),5.12(1H,d,J=6.4Hz),5.95(2H,s),6.77(1H,d,J=7.6Hz),6.87(1H,d,J=8.0Hz), 7.12(1H,s),7.24(1H,d,J=6.4Hz),7.45(1H,d,J=8.0Hz),8.31(1H,s),9.75(1H,s),12.33(1H,s).

[0184] Example 13

[0185] Preparation of 4-((2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ethynyl)cyclohexane-1-ol

[0186]

[0187] Step 1: Preparation of methyl 4-(tert-butyldiphenylsilyl)oxy)cyclohexane-1-carboxylate. methyl 4-hydroxycyclohexanecarboxylate (2.10 g, 13.28 mmol) was added to DCM (50 mL), followed by the addition of imidazole (0.90 g, 13.28 mmol) and TBDPSCl (3.65 g, 13.28 mmol). The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated to dryness, and the residue was purified by column chromatography (PE / EA = 100 / 1) to obtain the target compound (4.80 g, yield 91.3%) as a colorless oil.

[0188] EM (calculated value): 396.2; MS (ESI) m / z (M+H) + 397.2

[0189] Step 2: Preparation of 4-((tert-butyldiphenylsilyl)oxy)cyclohexane-1-carboxaldehyde

[0190] 1.00 g (2.52 mmol) of methyl 4-(tert-butyldiphenylsilyl)oxy)cyclohexane-1-carboxylic acid was added to anhydrous DCM (15 mL), and DIBAL-H (3.0 mL, 3.03 mmol, 1.0 M in n-hexane) was slowly added dropwise. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, water was added to the reaction solution and the mixture was extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying with anhydrous sodium sulfate, the reaction solution was concentrated to dryness to give the target compound (911 mg, yield 98.7%) as a colorless oil.

[0191] EM (calculated value): 366.2; MS (ESI) m / z (M+H) + 367.2

[0192] Step 3: Preparation of tert-butyl((4-ethynylcyclohexyl)oxy)diphenylsilane

[0193] Compound 4-((tert-butyldiphenylsilyl)oxy)cyclohexane-1-carboxaldehyde (800 mg, 2.18 mmol) was added to MeOH (15 mL), followed by the addition of potassium carbonate (752 mg, 5.45 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (837 mg, 4.36 mmol). The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction solution was concentrated to dryness, and the residue was purified by column chromatography (PE / EA = 80 / 1) to obtain the target compound (431 mg, yield 53.9%) as a colorless oil.

[0194] EM (calculated value): 362.2; MS (ESI) m / z (M+H) + 363.2

[0195] Step 4: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-6-(4-((tert-butyldiphenylsilyl)oxy)cyclohexyl)ethynyl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)quinazolin-4-amine)

[0196] Compounds 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (100 mg, 0.16 mmol) and tert-butyl((4-ethynylcyclohexyl)oxy)diphenylsilane (116 mg, 0.32 mmol) were added to DMF (5 mL), followed by CuI (4 mg, 0.02 mmol), Et3N (48 mg, 0.48 mmol), and Pd(PPh3)2Cl2 (22 mg, 0.03 mmol). The mixture was stirred at 45 °C for 4 hours under nitrogen protection. After the reaction was complete, water (50 mL) was added to the reaction solution, and the mixture was extracted three times with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (67 mg, yield 49.4%), which was a yellow oil.

[0197] EM (calculated value): 847.4; MS (ESI) m / z (M+H) + : 848.5

[0198] Step 5: Preparation of 4-((2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ethynyl)cyclohexane-1-ol

[0199] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-6-(4-((tert-butyldiphenylsilyl)oxy)cyclohexyl)ethynyl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)quinazolin-4-amine (67 mg, 0.08 mmol) was dissolved in THF (5 mL), and TBAF·3H2O (76 mg, 0.24 mmol) was added. The mixture was stirred overnight at room temperature. After the reaction was complete, water (20 mL) was added to the reaction solution, and the mixture was extracted three times with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying with anhydrous sodium sulfate, the solution was concentrated to dryness, and the crude product was purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (13 mg, yield 26.7%) as a pale yellow solid.

[0200] EM (calculated value): 609.3; MS (ESI) m / z (M+H) + : 610.4

[0201] 1 H NMR(400MHz,DMSO-d6)δ0.76-0.80(2H,m),0.93-0.97(2H,m),1.39-1.47(2H,m),1.52-1.58(2H,m) ,1.59-1.65(1H,m),1.74-1.81(1H,m),1.83-1.87(4H,m),1.94-2.01(4H,m),3.40-3.47(4H,m),4. 28-4.39(1H,m),4.57(1H,d,J=4.0Hz),5.96(2H,s),6.881(1H,d,J=8.0Hz),6.94(1H,d,J=8.0Hz), 7.13(1H,s),7.24(1H,d,J=8.0Hz),7.46(1H,d,J=8.0Hz),8.32(1H,s),9.76(1H,s),12.28(1H,s).

[0202] Example 14

[0203] Preparation of (4-((2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ynyl)-2-oxehedgeane[2.2.2]octane-1-methyl)methanol

[0204]

[0205] Step 1: Preparation of 1-((tert-butyldiphenylsilyl)oxy)methyl)-N-methoxy-N-methyl-2-oxeheno[2.2.2]octane-4-carboxamide

[0206] Compound 1-((tert-butyldiphenylsilyl)oxy)methyl)-2-oxobicyclo[2.2.2]octane-4-carboxylic acid (1.00 g, 2.36 mmol) was added to DCM (30 mL), followed by the addition of dimethylhydroxylamine hydrochloride (276 mg, 2.83 mmol), HBTU (1.34 g, 3.54 mmol), and DIEA (760 mg, 5.90 mmol). The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction solution was concentrated to dryness, and the residue was purified by column chromatography (PE / EA = 50 / 1) to obtain the target compound (708 mg, yield 64.2%) as a colorless oil.

[0207] EM (calculated value): 467.2; MS (ESI) m / z (M+H) + : 468.2

[0208] Step 2: Preparation of 1-((tert-butyldiphenylsilyl)oxy)methyl)-2-oxeheno[2.2.2]octane-4-carboxaldehyde

[0209] Compound 1-((tert-butyldiphenylsilyl)oxy)methyl)-N-methoxy-N-methyl-2-oxeheno[2.2.2]octane-4-carboxamide (700 mg, 1.50 mmol) was added to anhydrous THF (10 mL), cooled in an ice-water bath, and then DIBAL-H (2.3 mL, 2.3 mmol, 1.0 M in n-hexane) was slowly added dropwise. The mixture was then stirred at room temperature for 3 hours. After the reaction was complete, water was added to the reaction solution and the mixture was extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying with anhydrous sodium sulfate, the reaction solution was concentrated to dryness to give the target compound (435 mg, yield 71.0%) as a colorless oil.

[0210] EM (calculated value): 408.2; MS (ESI) m / z (M+H) + : 409.2

[0211] Step 3: Preparation of tert-butyl((4-ethynyl-2-oxeheno[2.2.2]octane-1-yl)methoxy)diphenylsilane

[0212] Compound 1-((tert-butyldiphenylsilyl)oxy)methyl)-2-oxeheno[2.2.2]octane-4-carboxaldehyde (435 mg, 1.07 mmol) was added to MeOH (10 mL), followed by the addition of potassium carbonate (443 mg, 3.21 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (411 mg, 2.14 mmol). The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction solution was concentrated to dryness, and the residue was purified by column chromatography (PE / EA = 50 / 1) to obtain the target compound (386 mg, yield 89.4%) as a colorless oil.

[0213] EM (calculated value): 404.2; MS (ESI) m / z (M+H) + : 405.2

[0214] Step 4: Preparation of (4-ethynyl-2-oxeheno[2.2.2]octane-1-yl)methanol

[0215] The compound tert-butyl((4-ethynyl-2-oxehera[2.2.2]octane-1-yl)methoxy)diphenylsilane (386 mg, 0.95 mmol) was dissolved in THF (10 mL), and TBAF·3H2O (899 mg, 2.85 mmol) was added. The mixture was stirred overnight at room temperature. After the reaction was complete, water (30 mL) was added to the reaction solution, and the mixture was extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying with anhydrous sodium sulfate, the solution was concentrated to dryness to obtain the target compound (359 mg, crude product) as a colorless oil.

[0216] EM (calculated value): 166.1; MS (ESI) m / z (M+H) + : 167.1;

[0217] Step 5: Preparation of (4-((2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ynyl)-2-oxehenocyclic[2.2.2]octane-1-methyl)methanol

[0218] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (100 mg, 0.16 mmol) and (4-ethynyl-2-oxehericyclo[2.2.2]octane-1-yl)methanol (359 mg, crude) were added to DMF (10 mL). CuI (4 mg, 0.02 mmol), Et3N (48 mg, 0.48 mmol), and Pd(PPh3)2Cl2 (22 mg, 0.03 mmol) were added to the mixture, and the mixture was stirred at 45 °C for 3 hours under nitrogen protection. After the reaction was complete, water (100 mL) was added to the reaction solution, and the mixture was extracted three times with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (62 mg, yield 59.6%) as a pale yellow solid.

[0219] EM (calculated value): 651.3; MS (ESI) m / z (M+H) + : 652.4;

[0220] 1 H NMR(400MHz,DMSO-d6)δ0.76-0.80(2H,m),0.93-0.97(2H,m),1.39-1.47(2H,m),1.52-1.58(2H,m) ,1.59-1.65(1H,m),1.74-1.81(1H,m),1.83-1.87(4H,m),1.94-2.01(4H,m),3.40-3.47(4H,m),4. 28-4.39(1H,m),4.57(1H,d,J=4.0Hz),5.96(2H,s),6.881(1H,d,J=8.0Hz),6.94(1H,d,J=8.0Hz), 7.13(1H,s),7.24(1H,d,J=8.0Hz),7.46(1H,d,J=8.0Hz),8.32(1H,s),9.76(1H,s),12.28(1H,s).

[0221] Example 15

[0222] Preparation of 4-((2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ethynyl)tetrahydro-2H-pyran-3-ol

[0223]

[0224] Step 1: Preparation of 3,7-dioxane[4.1.0]heptane

[0225] Compound 3,6-dihydro-2H-pyran (1.00 g, 11.89 mmol) was added to DCM (20 mL), cooled to 0 °C in an ice-water bath, and then m-CPBA (4.10 g, 23.78 mmol) was added in portions. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction solution was washed with water, the organic phase was collected and washed once with saturated NaCl aqueous solution, dried over anhydrous sodium sulfate, and then concentrated to dryness. The residue was purified by column chromatography (PE / EA = 20 / 1) to give the target compound (600 mg, yield 50.4%) as a colorless oil.

[0226] EM (calculated value): 100.1; MS (ESI) m / z (M+H) + : 101.1

[0227] Step 2: Preparation of 4-((trimethylsilyl)ethynyl)tetrahydro-2H-pyran-3-ol

[0228] Trimethylsilylacetylene (1.18 g, 11.98 mmol) was added to anhydrous THF (10 mL), cooled to -78 °C, and n-BuLi (7.20 mL, 14.38 mmol, 2 M in n-hexane) was slowly added. After stirring for 1 hour, 3,7-dioxane[4.1.0]heptane (600 mg, 5.99 mmol) was slowly added dropwise, and the temperature was maintained while stirring continued for 0.5 hours. Boron trifluoride diethyl ether solution was then added, and stirring continued for another 0.5 hours. After the reaction was complete, saturated NH4Cl aqueous solution was slowly added to quench the reaction mixture, followed by the addition of water (20 mL) and EA (20 mL) and stirring. The mixture was separated, and the aqueous phase was extracted once with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain the target compound (465 mg, yield 39.2%) as a yellow oil.

[0229] EM (calculated value): 198.1; MS (ESI) m / z (M+H) + 199.1

[0230] Step 3: Preparation of 4-ethyltetrahydro-2H-pyran-3-ol

[0231] Compound 4-((trimethylsilyl)ethynyl)tetrahydro-2H-pyran-3-ol (465 mg, 2.35 mmol) was added to MeOH (15 mL), followed by potassium carbonate (649 mg, 4.70 mmol). The mixture was stirred at room temperature for 0.5 hours. After the reaction was complete, the reaction solution was concentrated to dryness, and the residue was purified by column chromatography (PE / EA = 15 / 1) to obtain the target compound (236 mg, yield 79.7%) as a yellow oil.

[0232] EM (calculated value): 126.1; MS (ESI) m / z (M+H) + : 127.2

[0233] Step 4: Preparation of 4-((2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ethynyl)tetrahydro-2H-pyran-3-ol

[0234] Compounds 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (100 mg, 0.16 mmol) and 4-ethyltetrahydro-2H-pyran-3-ol (40 mg, 0.32 mmol) were added to DMF (3 mL), followed by CuI (4 mg, 0.02 mmol), Et3N (48 mg, 0.48 mmol), and Pd(PPh3)2Cl2 (22 mg, 0.03 mmol). The mixture was stirred at 45 °C for 4 hours under nitrogen protection. After the reaction was complete, water (100 mL) was added to the reaction solution, and the mixture was extracted three times with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (16 mg, yield 16.4%) as a yellow solid.

[0235] EM (calculated value): 611.3; MS (ESI) m / z (M+H) + : 612.4

[0236] 1H NMR(400MHz,DMSO-d6)δ0.77-0.79(2H,m),0.95-1.00(2H,m),1.22-1.25(2H,m),1.60-1.63(2H,m),1.8 6-1.89(2H,m),2.01-2.04(1H,m),2.65-2.70(1H,m),3.07-3.12(1H,m),3.49-3.55(4H,m),3.77-3.84( 2H,m),4.18-4.25(2H,m),5.26(1H,d,J=8.0Hz),5.98(2H,s),6.84(1H,d,J=8.0Hz),6.97(1H,d,J=8.0H z),7.15(1H,s),7.27(1H,d,J=8.0Hz),7.53(1H,d,J=12.0Hz),8.37(1H,s),9.82(1H,s),12.30(1H,s).

[0237] Example 16

[0238] Preparation of (5-((2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ethynyl)tetrahydro-2H-pyran-2-yl)methanol

[0239]

[0240] Step 1: Preparation of 6-(((tert-butyldiphenylsilyl)oxy)methyl)-N-methoxy-N-methyltetrahydro-2H-pyran-3-carboxamide

[0241] Compound 6-((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-carboxylic acid (556 mg, 1.40 mmol) was added to DCM (20 mL), followed by the addition of dimethylhydroxylamine hydrochloride (205 mg, 2.10 mmol), HBTU (637 mg, 1.68 mmol), and DIEA (452 ​​mg, 3.50 mmol). The mixture was stirred at room temperature for 5 hours. After the reaction was complete, the reaction solution was concentrated to dryness, and the residue was purified by column chromatography (PE / EA = 10 / 1) to give the target compound (524 mg, yield 84.8%) as a colorless oil.

[0242] EM (calculated value): 441.2; MS (ESI) m / z (M+H) + : 442.2

[0243] Step 2: Preparation of 6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-carboxaldehyde

[0244] Compound 6-(((tert-butyldiphenylsilyl)oxy)methyl)-N-methoxy-N-methyltetrahydro-2H-pyran-3-carboxamide (524 mg, 1.19 mmol) was added to anhydrous THF (10 mL). After cooling in an ice-water bath, DIBAL-H (2.4 mL, 2.38 mmol, 1.0 M in n-hexane) was slowly added dropwise, and the mixture was stirred for 0.5 hours while maintaining the temperature. After the reaction was complete, water was added to the reaction solution and the mixture was extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying with anhydrous sodium sulfate, the reaction solution was concentrated to dryness to obtain the target compound (1.20 g, crude product) as a colorless oil.

[0245] EM (calculated value): 382.2; MS (ESI) m / z (M+H) + 383.3

[0246] Step 3: Preparation of tert-butyl((5-ethyltetrahydro-2H-pyran-2-ynyl)methoxy)diphenylsilane

[0247] Compound 6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-carboxaldehyde (1.20 g, crude) was added to MeOH (10 mL), followed by the addition of potassium carbonate (493 mg, 3.57 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (419 mg, 2.38 mmol). The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction solution was concentrated to dryness, and the residue was purified by column chromatography (PE / EA = 20 / 1) to obtain the target compound (285 mg, overall yield of the two-step reaction was 63.3%) as a colorless oil.

[0248] EM (calculated value): 378.2; MS (ESI) m / z (M+H) + 379.3

[0249] Step 4: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-6-(6-((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)ethynyl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-methyl)quinazolin-4-amine)

[0250] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (227 mg, 0.37 mmol) and tert-butyl((5-ethyltetrahydro-2H-pyran-2-ynyl)methoxy)diphenylsilane (285 mg, 0.75 mmol) were added to DMF (10 mL), followed by CuI (8 mg, 0.04 mmol), Et3N (112 mg, 1.11 mmol), and Pd(PPh3)2Cl2 (28 mg, 0.04 mmol). The mixture was stirred at 45 °C for 4 hours under nitrogen protection. After the reaction was complete, water (100 mL) was added to the reaction solution, and the mixture was extracted three times with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 20 / 1) to obtain the target compound (200 mg, yield 62.7%), which was a brownish-yellow oil.

[0251] EM (calculated value): 863.4; MS (ESI) m / z (M+H) + : 864.5

[0252] Step 5: Preparation of (5-((2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ethynyl)tetrahydro-2H-pyran-2-yl)methanol

[0253] 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-6-(6-((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)ethynyl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-methyl)quinazolin-4-amine (200 mg, 0.23 mmol) was dissolved in THF (10 mL), and TBAF·3H₂O (218 mg, 0.69 mmol) was added. The mixture was stirred at 40 °C for 2 hours. After the reaction was complete, water (30 mL) was added to the reaction solution, and the mixture was extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 20 / 1) to obtain the target compound (97 mg, yield 67.5%) as a deep yellow solid.

[0254] EM (calculated value): 625.3; MS (ESI) m / z (M+H) + : 626.4

[0255] 1 H NMR(400MHz,DMSO-d6)δ0.77-0.80(2H,m),0.94-0.96(2H,m),1.28-1.34(1H,m),1.57-1.67(4 H,m),1.84-1.89(3H,m),2.11-2.20(1H,m),3.48-3.63(6H,m),4.02-4.07(1H,m),4.13-4.37( 2H,m),4.67(1H,t,J=4.0Hz),5.97(2H,s),6.82(1H,d,J=8.0Hz),6.95(1H,d,J=12.0Hz),7.14 (1H,s),7.25(1H,d,J=8.0Hz),7.48(1H,d,J=8.0Hz),8.34(1H,s),9.80(1H,s),12.30(1H,s).

[0256] Example 17

[0257] Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-(1H-pyrazol-4-yl)quinazolin-4-amine

[0258]

[0259] Compounds 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (30 mg, 0.05 mmol) and pinacol 4-pyrazolboronic acid (19 mg, 0.10 mmol) were added to 1,4-dioxane / H2O (2 mL, 5 / 1). Sodium carbonate (21 mg, 0.20 mmol) and Pd(dppf)Cl2·DCM (7 mg, 0.01 mmol) were added to the mixture. The mixture was stirred at 80 °C for 4 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness. The crude product was purified by column chromatography (DCM / MeOH / NH3·H2O = 20 / 1 / 0.05) to obtain the target compound (5 mg, yield 18.1%) as a white solid.

[0260] EM (calculated value): 553.2; MS (ESI) m / z (M+H) + 554.3

[0261] 1H NMR(400MHz,DMSO-d6)δ0.77-0.80(2H,m),0.95-0.96(2H,m),1.54-1.57(2H,m),1.82- 1.91(4H,m),1.96-2.01(1H,m),3.41-3.50(2H,m),4.30-4.34(2H,m),5.96(2H,s),6.80 (1H,d,J=7.6Hz),6.94(1H,d,J=9.6Hz),7.13(1H,s),7.32(1H,d,J=8.4Hz),7.85(1H,d ,J=8.8Hz),8.01-8.15(2H,m),8.48(1H,s),9.72(1H,s),12.32(1H,s),12.94(1H,brs).

[0262] Example 18

[0263] Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-(1-methyl-1H-pyrazol-4-yl)quinazolin-4-amine

[0264]

[0265] Compounds 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (30 mg, 0.05 mmol) and pinacol ester of 1-methyl-4-pyrazolboronic acid (21 mg, 0.10 mmol) were added to 1,4-dioxane / H2O (2 mL, 5 / 1), followed by the addition of sodium carbonate (21 mg, 0.20 mmol) and Pd(dppf)Cl2.DCM (7 mg, 0.01 mmol). The mixture was stirred at 80 °C for 4 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness, and the crude product was purified by column chromatography (DCM / MeOH / NH3.H2O=20 / 1 / 0.05) to obtain the target compound (4 mg, yield 14.1%) as a white solid.

[0266] EM (calculated value): 567.3; MS (ESI) m / z (M+H) + 568.3

[0267] 1H NMR(400MHz,DMSO-d6)δ0.74-0.77(2H,m),0.93-0.95(2H,m),1.44(3H,s),1.50-1.55(2H,m ),1.78-1.86(2H,m),1.87-1.89(2H,brs),1.91-1.96(1H,m),3.45-3.52(2H,m),4.33-4.37 (2H,m),5.95(2H,s),6.77(1H,d,J=6.4Hz),6.91(1H,d,J=8.0Hz),7.12(1H,s),7.30(1H,d, J=7.2Hz),7.80(1H,d,J=8.0Hz),8.00-8.11(2H,m),8.44(1H,s),9.72(1H,s),12.32(1H,s).

[0268] Example 19

[0269] Preparation of 3-(4-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)phenoxy)propane-1,2-diol

[0270]

[0271] Step 1: Preparation of 2-(4-((2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)pinacol ester of phenylboronic acid

[0272] The compound 4-hydroxyphenylboronic acid pinacol ester (2.20 g, 10.00 mmol) was added to DMSO (40 mL), followed by the addition of potassium carbonate (2.07 g, 15.00 mmol) and 4-(chloromethyl)-2,2-dimethyl-1,3-dioxolane (1.80 g, 12.00 mmol). The mixture was heated to 120 °C and stirred for 10 hours. After the reaction was complete, the reaction solution was cooled to room temperature, and water (400 mL) was added. The mixture was extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying with anhydrous sodium sulfate, the solution was concentrated to dryness. The residue was purified by column chromatography (PE / EA = 10 / 1) to obtain the target compound (1.7 g, yield 50.9%) as a brown oil.

[0273] EM (calculated value): 334.2; MS (ESI) m / z (M+H) + 335.2

[0274] Step 2: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-(4-((2,2-dimethyl-1,3-dioxo-4-yl)methoxy)phenyl)quinazolin-4-amine)

[0275] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (100 mg, 0.16 mmol) and 2-(4-((2,2-dimethyl-1,3-dioxolane-4-yl)methoxy)pinacol ester of phenylboronic acid (107 mg, 0.32 mmol) were added to 1,4-dioxan In a mixture of e / H2O (10 mL, 5 / 1), sodium carbonate (34 mg, 0.32 mmol) and Pd(dppf)Cl2·DCM (14 mg, 0.02 mmol) were added, and the mixture was stirred at 80 °C for 5 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness, and the crude product was purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (50 mg, yield 45.1%) as a brown solid.

[0276] EM (calculated value): 693.3; MS (ESI) m / z (M+H) + : 694.4

[0277] Step 3: Preparation of 3-(4-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)phenoxy)propane-1,2-diol)

[0278] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-(4-((2,2-dimethyl-1,3-dioxocyclo-4-yl)methoxy)phenyl)quinazolin-4-amine (50 mg, 0.07 mmol) was added to THF (5 mL), followed by the addition of concentrated hydrochloric acid (0.5 mL). The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated to dryness, and the crude product was purified by column chromatography (DCM / MeOH = 15 / 1) to obtain the target compound (20 mg, yield 42.5%) as a pale yellow solid.

[0279] EM (calculated value): 653.3; MS (ESI) m / z (M+H) + : 654.4

[0280] 1 H NMR(400MHz,DMSO-d6)δ0.76-0.79(2H,m),0.83-0.85(2H,m),1.53-1.56(2H,m),1.79-1.84(2H,m),1.87-1.89(1H,m), 1.92-2.01(2H,brs),3.45-3.48(2H,m),3.81-3.82(1H,m),3.89-3.93(1H,m),4.03-4.07(1H,m),4.31-4.39(2H,m),4. 70-4.73(1H,m),4.99-5.00(1H,m),5.95(2H,s),6.80(1H,d,J=8.0Hz),6.92(1H,d,J=7.6Hz),7.04(2H,d,J=8.8Hz),7. 13(1H,s),7.37(1H,d,J=8.8Hz),7.75(2H,d,J=8.8Hz),7.90(1H,d,J=8.8Hz),8.54(1H,s),9.91(1H,s),12.30(1H,s).

[0281] Example 20

[0282] Preparation of 1-(4-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)phenyl)ethane-1,2-diol)

[0283]

[0284] Step 1: Preparation of 1-(4-bromophenyl)ethane-1,2-diol

[0285] 1-Bromo-4-vinylbenzene (200 mg, 1.10 mmol) was added to DCM / H2O (10 mL, 4 / 1), followed by NMO (258 mg, 2.20 mmol) and KOsO4·2H2O (810 mg, 2.20 mmol). The mixture was stirred at room temperature for 3 hours. After the reaction was complete, the reaction solution was concentrated to dryness. The crude product was purified by column chromatography (PE / EA = 5 / 1) to obtain the target compound (120 mg, yield 50.5%) as a white solid.

[0286] EM (calculated value): 216.0; MS (ESI) m / z (M+H) + : 217.0

[0287] Step 2: Preparation of 1-(4-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)phenyl)ethane-1,2-diol)

[0288] The compound 1-(4-bromophenyl)ethane-1,2-diol (100 mg, 0.46 mmol) was dissolved in 1,4-dioxane (5 mL), and potassium acetate (90 mg, 0.92 mmol) and Pd(dppf)Cl2.DCM (28 mg, 0.04 mmol) were added sequentially. The mixture was stirred at 90 °C for 5 hours under nitrogen protection. After the reaction was complete, the reaction solution was cooled to room temperature, and 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (140 mg, 0.23 mmol), sodium carbonate (49 mg, 0.46 mmol), and Pd(dppf)Cl2·DCM (14 mg, 0.02 mmol) were added sequentially. The mixture was stirred at 90 °C for 6 hours under nitrogen protection. After the reaction was complete, the reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated to dryness. The crude product was purified sequentially by column chromatography and preparative HPLC to obtain the target compound (5 mg, yield 3.5%) as a yellow solid.

[0289] EM (calculated value): 623.3; MS (ESI) m / z (M+H) + : 624.4

[0290] 1 H NMR(400MHz,DMSO-d6)δ0.78-0.81(2H,m),0.93-0.98(2H,m),1.51-1.54(2H,m),1.76-1.80(2H,m), 1.81-1.84(1H,m),1.89-1.95(2H,brs),3.41-3.47(2H,m),4.13-4.18(2H,m),4.25-4.32(2H,m),5. 00-5.06(1H,m),5.18-5.27(1H,m),5.55-5.60(1H,m),5.96(2H,s),6.81-6.86(2H,m),6.92-6.98(2 H,m),7.13(1H,s),7.33(2H,d,J=6.4Hz),7.52-7.58(2H,m),8.41(1H,s),9.87(1H,s),12.33(1H,s).

[0291] Example 21

[0292] Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-vinylquinazoline-4-amine

[0293]

[0294] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (60 mg, 0.10 mmol) and potassium ethylene trifluoroborate (27 mg, 0.20 mmol) were added to 1,4-dioxane / H2O (10 mL, 5 / 1). Et3N (30 mg, 0.30 mmol) and Pd(dppf)Cl2·DCM (14 mg, 0.02 mmol) were then added. The mixture was stirred at 80 °C for 4 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness. The crude product was purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (46 mg, yield 89.6%) as a yellow solid.

[0295] EM (calculated value): 513.2; MS (ESI) m / z (M+H) + 514.3

[0296] 1 H NMR(400MHz,DMSO-d6)δ0.78-0.81(2H,m),0.94-0.97(2H,m),1.51-1.54(2H,m),1.72-1.77( 2H,m),1.85-1.88(1H,m),1.90-1.95(2H,brs),3.37-3.43(2H,m),4.25-4.35(2H,m),5.25-5. 351(2H,m),5.95(2H,s),6.02-6.05(1H,m),6.83(1H,d,J=6.4Hz),6.90(1H,d,J=8.0Hz),7.13 (1H,s),7.29(1H,d,J=8.0Hz),7.51(1H,d,J=8.4Hz),8.37(1H,s),9.81(1H,s),12.30(1H,s).

[0297] Example 22

[0298] Preparation of 1-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)ethane-1,2-diol

[0299]

[0300] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-vinylquinazoline-4-amine (40 mg, 0.08 mmol) was added to DCM / THF / H2O (10 mL, 4 / 1 / 1), followed by the addition of NMO (19 mg, 0.16 mmol) and KOsO4·2H2O (60 mg, 0.16 mmol). The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was concentrated to dryness, and the crude product was purified by preparative HPLC to obtain the target compound (10 mg, yield 22.9%) as a yellow solid.

[0301] EM (calculated value): 547.2; MS (ESI) m / z (M+H) + 548.3

[0302] 1 H NMR(400MHz,DMSO-d6)δ0.75-0.79(2H,m),0.91-0.94(2H,m),1.49-1.52(2H,m),1.75-1.79(2H,m ),1.89-1.96(3H,m),3.39-3.44(2H,m),4.11-4.17(2H,m),4.27-4.34(2H,m),5.02-5.09(1H,m), 5.20-5.29(1H,m),5.59-5.63(1H,m),5.97(2H,s),6.87(1H,d,J=5.2Hz),6.94(1H,d,J=7.6Hz),7 .12(1H,s),7.33(1H,d,J=7.6Hz),7.55(1H,d,J=8.0Hz),8.39(1H,s),9.84(1H,s),12.31(1H,s).

[0303] Example 23

[0304] Preparation of (5-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)-3,4-dihydro-2H-pyran-2-yl)methanol

[0305]

[0306] Step 1: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-6-(2-((tert-butyldiphenylsilyl)oxy)methyl)-3,4-dihydro-2H-pyran-5-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)quinazolin-4-amine

[0307] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (120 mg, 0.20 mmol) and tert-butyldiphenyl((5-(4,4,5,5-tetramethyl-1,3,2-dioxobenzofuran-2-yl)-3,4-dihydro-2H-pyran-2-yl)methoxy)silane (120 mg, 0.25 mmol) were added to 1,4-dioxane / H2O (10 mL, 5 / 1), and sodium carbonate (42 mg, 0.40 mmol) and Pd(dppf)Cl2.DCM (21 mg, 0.03 mmol) were added to the mixture. The mixture was stirred overnight at 80 °C under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness, and the crude product was purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (80 mg, yield 47.8%) as a yellow solid.

[0308] EM (calculated value): 837.4; MS (ESI) m / z (M+H) + : 838.5

[0309] Step 2: Preparation of (5-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)-3,4-dihydro-2H-pyran-2-yl)methanol

[0310] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-6-(2-((tert-butyldiphenylsilyl)oxy)methyl)-3,4-dihydro-2H-pyran-5-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)quinazolin-4-amine (80 mg, 0.10 mmol) was dissolved in THF (10 mL), and TBAF·3H₂O (95 mg, 0.30 mmol) was added. The mixture was stirred at 40 °C for 3 hours. After the reaction was complete, water (30 mL) was added to the reaction solution, and the mixture was extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 20 / 1) to obtain the target compound (50 mg, yield 83.3%) as a white solid.

[0311] EM (calculated value): 599.3; MS (ESI) m / z (M+H) + 600.4

[0312] 1 H NMR(400MHz,DMSO-d6)δ0.81-0.85(2H,m),0.96-1.00(2H,m),1.43-1.47(2H,m),1.74-1.78(2H,m),1.88- 1.92(1H,m),1.99-2.04(2H,m),2.13-2.17(2H,m),3.45-3.52(2H,m),3.79-3.86(2H,m),4.46-4.50(1H,m) ,4.68-4.77(2H,m),5.33(1H,t,J=4.0Hz),6.02(2H,s),6.38(1H,s),6.92(1H,d,J=8.0Hz),7.04(1H,d,J=8 .0Hz),7.21(1H,s),7.32(1H,d,J=8.0Hz),7.72(1H,d,J=12.0Hz),8.12(1H,s),9.87(1H,s),12.34(1H,s).

[0313] Example 24

[0314] Preparation of (5-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)tetrahydro-2H-pyran-2-yl)methanol

[0315]

[0316] The compound (5-(2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-yl)-3,4-dihydro-2H-pyran-2-yl)methanol (40 mg, 0.07 mmol) was dissolved in 10 mL of methanol. Pd / C (10 mg, 10% purity) was added, and the mixture was purged with hydrogen and stirred overnight at room temperature. After the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated to dryness. The crude product was purified by column chromatography (DCM / MeOH = 20 / 1) to obtain the target compound (20 mg, yield 47.6%) as a white solid.

[0317] EM (calculated value): 601.3; MS (ESI) m / z (M+H) + 602.4

[0318] 1 H NMR(400MHz,DMSO-d6)δ0.79-0.82(2H,m),0.93-0.95(2H,m),1.37-1.42(2H,m),1.70-1.784(2H,m),1.85-1.90( 1H,m),1.79-2.03(2H,m),2.11-2.16(2H,m),2.35-2.39(1H,m),3.47-3.53(2H,m),3.81-3.87(2H,m),4.49-4.53 (1H,m),4.56-4.60(2H,m),4.69-4.76(2H,m),5.34(1H,t,J=4.0Hz),5.96(2H,s),6.88(1H,d,J=7.6Hz),7.05(1H ,d,J=8.4Hz),7.15(1H,s),7.28(1H,d,J=6.4Hz),7.68(1H,d,J=8.0Hz),8.21(1H,s),9.69(1H,s),12.28(1H,s).

[0319] Example 25

[0320] Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)-N-(1-(hydroxymethyl)-2-oxehezoryl[2.2.2]octane-4-yl)quinazolin-6-carboxamide

[0321]

[0322] Step 1: Preparation of 1-(((tert-butyldiphenylsilyl)oxy)methyl)-2-oxehenocyclic[2.2.2]octane-4-amine

[0323] Compound 1-((tert-butyldiphenylsilyl)oxy)methyl)-2-oxobicyclo[2.2.2]octane-4-carboxylic acid (1.00 g, 2.36 mmol) was added to toluene (15 mL), followed by Et3N (715 mg, 7.08 mmol), molecular sieve, and DPPA (1.30 g, 4.72 mmol). The mixture was heated to 100 °C and stirred for 4 hours. After the reaction was complete, the reaction solution was concentrated to dryness, and the residue was dissolved in THF (20 mL). Potassium tert-butoxide (530 mg, 4.72 mmol) was added, and the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction solution was concentrated to dryness, and the crude product was purified by column chromatography (PE / EA = 5 / 1) to obtain the target compound (437 mg, yield 46.9%) as a yellow oil.

[0324] EM (calculated value): 395.2; MS (ESI) m / z (M+H) + 396.2

[0325] Step 2: Preparation of methyl 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-carboxylic acid ester

[0326] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)-6-iodoquinazolin-4-amine (75 mg, 0.12 mmol) was added to methanol (10 mL), followed by the addition of Et3N (36 mg, 0.36 mmol) and Pd(dppf)Cl2·DCM (14 mg, 0.02 mmol). The mixture was stirred overnight at 70 °C under a carbon monoxide atmosphere. After the reaction was complete, the reaction solution was concentrated to dryness, and the crude product was purified by column chromatography (DCM / MeOH = 30 / 1) to obtain the target compound (63 mg, yield 96.3%) as a yellow solid.

[0327] EM (calculated value): 545.2; MS (ESI) m / z (M+H) + 546.3

[0328] Step 3: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-carboxylic acid

[0329] The compound methyl 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazoline-6-carboxylic acid ester (63 mg, 0.12 mmol) was added to methanol / H2O (10 mL, 4 / 1), and LiOH·H2O (15 mg, 0.36 mmol) was added. The mixture was heated to 50 °C and stirred for 2 hours. After the reaction was complete, the reaction solution was concentrated to dryness to obtain the target compound (crude product), which was a yellow solid.

[0330] EM (calculated value): 531.2; MS (ESI) m / z (MH) - 530.2

[0331] Step 4: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(1-((tert-butyldiphenylsilyl)oxy)methyl)-2-oxeheno[2.2.2]octane-4-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-carboxamide

[0332] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-carboxylic acid (crude) was added to THF (10 mL), followed by the sequential addition of 1-(((tert-butyldiphenylsilyl)oxy)methyl)-2-oxeheno[2.2.2]octane-4-amine (71 mg, 0.18 mmol), HATU (68 mg, 0.18 mmol), and DIEA (31 mg, 0.24 mmol). The mixture was stirred at room temperature for 4 hours. After the reaction was complete, the reaction solution was concentrated to dryness, and the residue was purified by column chromatography (DCM / MeOH = 20 / 1) to obtain the target compound (61 mg, overall yield of the two-step reaction was 56.0%) as a yellow solid.

[0333] EM (calculated value): 908.4; MS (ESI) m / z (M+H) + 909.5

[0334] Step 5: Preparation of 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-4-(5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)-N-(1-(hydroxymethyl)-2-oxehezoryl[2.2.2]octane-4-yl)quinazolin-6-carboxamide

[0335] Compound 2-(4-amino-4-(benzo[d][1,3]dioxo-5-yl)piperidin-1-yl)-N-(1-((tert-butyldiphenylsilyl)oxy)methyl)-2-oxetane[2.2.2]octane-4-yl)-4-((5-cyclopropyl-4-fluoro-1H-pyrazol-3-yl)amino)quinazolin-6-carboxamide (61 mg, 0.07 mmol) was dissolved in THF (10 mL), and TBAF·3H2O (66 mg, 0.21 mmol) was added. The mixture was stirred at 50 °C for 2 hours. After the reaction was complete, water (30 mL) was added to the reaction solution, and the mixture was extracted twice with EA. The organic phases were combined and washed once with saturated NaCl aqueous solution. After drying anhydrous sodium sulfate, the product was concentrated to dryness. The crude product was then purified by column chromatography (DCM / MeOH = 20 / 1) to obtain the target compound (15 mg, yield 34.1%) as a yellow solid.

[0336] EM (calculated value): 670.3; MS (ESI) m / z (M+H) + : 671.4

[0337] 1 H NMR(400MHz,DMSO-d6)δ0.79-0.81(2H,m),0.95-0.97(2H,m),1.62-1.83(6H,m),1.87-1.91(1H,m),1. 94-2.04(4H,m),2.12-2.31(2H,m),3.18(2H,d,J=4.0Hz),3.44-3.48(4H,m),3.59-3.63(2H,m),4.01(2 H,s),4.57(1H,t,J=4.0Hz),6.00(2H,s),6.88(1H,d,J=8.0Hz),7.01(1H,d,J=8.0Hz),7.17(1H,s),7. 31(1H,d,J=8.0Hz),7.53-7.57(1H,m),7.92(1H,d,J=12.0Hz),8.65(1H,s),9.84(1H,s),12.33(1H,s).

[0338] Inhibitory effect of compound in Experiment Example 1 on kinase activity

[0339] 1: Test materials

[0340] PAK4 (Carna, No. 07-126), PAK2 (Carna, No. 07-124), Kinase substrate31 (Cisbio, No. 61ST2BLE), DMSO (Sigma, No. D0632), 384-well plate (Greiner, No. 784075), PF-3758309 (selleckchem, No. S709403)

[0341] 2: Experimental Methods

[0342] 2.1 Compound Preparation

[0343] The compound was received by the administrator, who dissolved the powder in 100% DMSO to prepare a 10mM storage solution, which was then stored in a nitrogen cabinet away from light.

[0344] 2.2 Kinase Response Process

[0345] (1) Prepare 1×Kinase buffer.

[0346] (2) Preparation of compound concentration gradients: The test compound was prepared at a concentration of 1000 nM and diluted 100 times to a final concentration of 100% DMSO in a 384 source plate. The compound was then diluted 3 times with Precision to obtain 10 concentrations. 250 nL of the 100-fold final concentration of the compound was transferred to the target 384-well plate using an Echo 550 dispenser.

[0347] (3) Prepare a kinase solution with a final concentration of 2.5 times using 1×Kinase buffer.

[0348] (4) Add 10 μL of kinase solution at 2.5 times the final concentration to the compound wells and the positive control wells respectively; add 10 μL of 1×Kinase buffer to the negative control wells.

[0349] (5) Centrifuge at 1000 rpm for 30 seconds, shake the reaction plate to mix, and incubate at room temperature for 10 minutes.

[0350] (6) Prepare a mixed solution of ATP and Kinase substrate 22 at a final concentration of 5 / 3 times using 1×Kinase buffer.

[0351] (7) Add 15 μL of a mixture of ATP and substrate at 5 / 3 times the final concentration to initiate the reaction.

[0352] (8) Centrifuge the 384-well plate at 1000 rpm for 30 seconds, shake to mix, and incubate at room temperature for 60 min.

[0353] (9) Add 30 μL of the stop detection solution to stop the kinase reaction, centrifuge at 1000 rpm for 30 seconds, and shake to mix.

[0354] (10) Read the conversion rate using Caliper EZ Reader.

[0355] 2.3 Data Analysis

[0356]

[0357] Where: Conversion%_sample is the conversion rate reading of the sample; Conversion%_min: the mean value of the negative control wells, representing the conversion rate reading of wells without enzyme activity; Conversion%_max: the mean value of the positive control wells, representing the conversion rate reading of wells without compound inhibition.

[0358] Fitted dose-response curve:

[0359] Plotting the logarithmic value of concentration on the X-axis and the percentage inhibition rate on the Y-axis, dose-response curves were fitted using the log(inhibitor) vs. response-variable slope function of GraphPad Prism 5 to derive the IC50 values ​​of each compound on enzyme activity. 50 Value. The calculation formula is:

[0360] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0361] The test results are shown in Table 2:

[0362] Table 2. Inhibitory activities of compounds against PAK4 and PAK2 kinases (IC50) 50 )

[0363]

[0364] Experimental Example 2: hERG Test for Detecting the Cardiotoxicity of Compounds

[0365] 1. Experimental Principle

[0366] 1.1 The hERG ion channel was stably expressed in HEK293 cells. After the hERG current stabilized, the magnitude of the hERG current before and after the application of different compound concentrations could be compared to determine the effect of the compound on the hERG ion channel.

[0367] 1.2 Regulations to be followed

[0368] This experiment did not follow GLP guidelines, but was conducted in accordance with Olive's Standard Operating Procedures (SOP) and relevant literature published in SCI journals.

[0369] 2. Experimental animals (cells / reagents)

[0370] HEK293 cells with steady-state expression of hERG ion channels.

[0371] 3. Test instruments

[0372] Patch clamp instrument: PC-505B

[0373] Micro-manipulation instrument: MP-225

[0374] Electrode drawing instrument: PC-10 (Narishige, Japan)

[0375] 4. Drug preparation

[0376] The test compound was stored at a concentration of 3 mM and dissolved in dimethyl sulfoxide (DMSO). It was then dissolved in extracellular fluid on the day of the test to prepare the required concentration.

[0377] Except for NaOH and KOH used in acid-base titrations, all compounds were purchased from Sigma (St. Louis, MO). The final concentrations of all tested compounds were prepared on the same day and then dissolved in extracellular fluid. The extracellular fluid (mM) was: NaCl, 137; KCl, 4; CaCl2, 1.8; MgCl2, 1; HEPES, 10; glucose, 10; pH 7.4 (NaOH titration).

[0378] All test and control compound solutions contained 0.3% DMSO. Intracellular solution (mM) was: KAspartate, 130; MgCl2, 5; EGTA, 5; HEPES, 10; Tris-ATP, 4; pH 7.2 (KOH titration).

[0379] 5. Test methods

[0380] Cells: All experiments were performed at room temperature. Each cell served as its own control.

[0381] Compound testing: All compounds were perfused using a gravity-based perfusion system. At least one cell was tested for each concentration. After the current stabilized (or for 5 minutes), the change in current before and after compound application was compared to calculate the compound's blocking effect.

[0382] Positive control: None

[0383] Electrophysiology: Cells were transferred to a perfusion tank and perfused with extracellular fluid. The intracellular fluid (mM) consisted of: KAspartate, 130; MgCl2, 5; EGTA, 5; HEPES, 10; Tris-ATP, 4; pH 7.2 (titrated with KOH). Intracellular fluid was stored in batches at -80°C and thawed on the day of the experiment. Electrodes were fabricated using PC-10 (Narishige, Japan). Whole-cell patch-clamp recording was performed, and noise was filtered at one-fifth of the sampling frequency.

[0384] Test Procedure and Results: Cells were clamped at -80 mV, then depolarized to 40 mV with a 4-second square wave, followed by hyperpolarization to -40 mV with a 2-second square wave to obtain the hERG tail current. This procedure was repeated every 20 seconds. The hERG tail current is the pure hERG current. The maximum current induced by the second square wave was detected, and after it stabilized, the test compound was perfused. Once the reaction stabilized, the blocking strength was calculated.

[0385] The test results are shown in Table 3:

[0386] Table 3. Inhibition rate (%) of hERG channel current of compounds (at 1µm concentration)

[0387]

[0388]

[0389] Using the same experimental conditions and methods, the hERG inhibition rate of some preferred compounds in patent WO2022033420 A1 was tested, and the results are shown in Table 4 below:

[0390] Table 4. Inhibition rate (%) of hERG channel current of some compounds in patent WO2022033420 A1 (at 1µm concentration)

[0391] Example number Inhibition rate Example number Inhibition rate 71 85.70％ 96 77.12％ 79 75.32％ 97 84.17％

[0392] A higher inhibition rate indicates a greater risk of hERG cardiotoxicity. Table 4 shows that the preferred compounds in patent WO2022033420 A1 have higher hERG inhibition rates than most of the compounds prepared in Examples 1-25 of this invention, indicating a greater risk of cardiotoxicity. In contrast, the compounds prepared by this invention, after structural optimization, exhibit lower hERG inhibition rates and a lower risk of cardiotoxicity.

[0393] Experimental Example 3: Stability Test of Compound in Liver Microsomes

[0394] 1: Materials and Methods

[0395] Buffer solution:

[0396] (1) 100mM potassium phosphate buffer, pH 7.4; (2) 10mM MgCl2.

[0397] Preparation of compound solutions:

[0398] (1) Preparation of 100 μM working solution: Take 5 μL of the stock solution (10 mM) of the test group or control group and dilute it with 495 μL of methanol to obtain a compound concentration of 100 μM (99% MeOH).

[0399] (2) Preparation of 10 μM working solution: Take 50 μL of 100 μM working solution and dilute it with 450 μL of 100 mM potassium phosphate buffer to obtain a compound concentration of 10 μM (9.9% MeOH).

[0400] Components of the NADPH (prototype coenzyme II) regeneration system (final concentration of isocitrate dehydrogenase in culture medium is 1.0 unit / mL):

[0401] β-Nicotinamide adenine dinucleotide phosphate, Supplier: Chem-impex International, Product No.: N00616

[0402] Preparation of liver microsomal solution (final concentration 0.5 mg protein / mL), types of liver microsomes are shown in Table 5:

[0403] Table 5

[0404]

[0405] Termination solution:

[0406] An ice-cold acetonitrile solution containing 100 ng / mL tolbutamide and 100 ng / mL labetalol as internal standards.

[0407] Operating steps:

[0408] (1) Except for the blank matrix plate wells, 10 μL of working solution of the test or control drug was added to each of the other wells (T0, T5, T10, T20, T30, T60 and NCF60).

[0409] (2) Dispense 80 μL / well microparticle solution onto each plate with Apricot and incubate the mixture of microparticle solution and compound at 37°C for about 10 minutes.

[0410] (3) Add 10 μL of 100 mM potassium phosphate buffer to NCF60 / well, incubate at 37°C, and start timer 1. The time is shown in Table 6.

[0411] Table 6

[0412]

[0413] (4) After preheating, use Apricot to dispense 10 μL / well of NADPH regeneration system onto each plate to start the reaction.

[0414] Table 7 Final concentrations of each component in the incubation medium

[0415]

[0416] (5) Incubate at 37℃, start timer 2, and the data are shown in Table 8.

[0417] Table 8

[0418]

[0419] (6) Add pre-cooled stop solution (containing 100 ng / mL of internal standard tolbutamide and 100 ng / mL of sulfadiazine) to each well to terminate the reaction.

[0420] (7) Then the sample plate is shaken on the trigger for about 10 minutes.

[0421] (8) Centrifuge the sample at 4000 rpm for 20 min at 4℃.

[0422] (9) Take another 96-well plate, add 300 μL of HPLC-grade water to each well, add 100 μL of the supernatant obtained by centrifugation to the corresponding well, mix the two and use them for LC / MS / MS detection.

[0423] Data Analysis:

[0424] t is calculated based on first-order elimination kinetics. 1 / 2 and CL int(mic) value

[0425] The first-order elimination kinetic equation is:

[0426]

[0427]

[0428]

[0429]

[0430]

[0431] The results of liver microsomal stability tests for some compounds are shown in Table 9:

[0432] Table 9. Liver microsomal stability data for some compounds.

[0433]

[0434] Experiment 4: Detection of the PK properties of the compound in rats

[0435] Male SD rats (purchased from Chengdu Dashuo Experimental Animal Co., Ltd.) were used. Pharmacokinetic studies were conducted on SD rats by single oral administration (50 mg / kg, 3 rats per group) of each test compound. The test compounds were prepared on the day of administration using 5% DMSO + 10% solubilizer + 85% saline, followed by vortexing for 2 min and sonication for 5 min to prepare the drug solution. Animals were fasted for 10-14 hours before oral administration and resumed feeding 4 hours after administration. Pharmacokinetic samples were collected via the jugular vein after oral and intravenous administration to SD rats at the following time points: before administration, and 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration. Three whole blood samples (approximately 0.2 mL) were collected at each time point and anticoagulated with heparin sodium. Blood samples were immediately placed on ice and centrifuged within 1 hour to separate the plasma (centrifugation conditions: 6800 rpm, 6 minutes, 2-8℃). The collected plasma was stored in a -80°C freezer before analysis.

[0436] The pharmacokinetic test results of some compounds of this invention are shown in Table 10 below:

[0437] Table 10 Pharmacokinetic test results of some compounds of the present invention

[0438]

[0439]

[0440] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A compound of a PAK4 kinase inhibitor, characterized in that, The compound has the structure of Formula III or Formula V, or a pharmaceutically acceptable salt, stereoisomer thereof: （Ⅲ） R 12 selected from -H; R 13 , R 14 independently selected from -H; R 15 selected from -OH, substituted or non-substituted C1-10 linear / branched alkyl, substituted or non-substituted cycloalkyl or heterocycloalkyl, wherein the H on the above groups can be substituted with -OH, -CF3, hydroxymethyl, methyl, ethyl, propyl, isopropyl, phenyl; said substituted or non-substituted cycloalkyl or heterocycloalkyl is , , , , , , represents the position where the alkynyl group can be attached; （Ⅴ） R 21 selected from -H; R 22 , R 23 independently selected from H; Y is selected from substituted or non-substituted C1-4 linear / branched alkyl, substituted or non-substituted C3-8 heterocycloalkyl, substituted or non-substituted C5-6 aryl or heteroaryl, amido; said substituents are selected from -OH, alkyl, alkoxy, hydroxyalkyl; said substituted or non-substituted C3-8 heterocycloalkyl is , , , , " indicates the position of attachment to Y; said R 24 is selected from -H, -OH, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl.

2. The compound of claim 1, wherein The compound has the following structure: 。 3. A pharmaceutical composition comprising the compound of claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer thereof.

4. Use of the compound of claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer thereof, or the pharmaceutical composition of claim 3 in the preparation of a medicament for treating a disease associated with the expression or activity of PAK4 kinase. The disease associated with the expression or activity of PAK4 kinase is selected from a cancer, a neurodegenerative disease or an immune system disease; the cancer is selected from breast cancer, mantle cell lymphoma, ovarian cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, hepatocellular carcinoma, glioma, endometrial cancer, melanoma, renal cancer, bladder cancer, biliary tract cancer, pancreatic cancer, lymphoma, hairy cell cancer, pharyngeal cancer, large bowel cancer, rectal cancer, brain and central nervous system cancer, genitourinary tract cancer, lung cancer, bone cancer, colon cancer, adenocarcinoma, follicular carcinoma, Hodgkin's leukemia, bronchus cancer, uterine corpus cancer, uterine cervix cancer, multiple myeloma, acute myelogenous leukemia, chronic myelogenous leukemia, lymphocytic leukemia, chronic lymphoid leukemia, myelogenous leukemia, non-Hodgkin's lymphoma, primary macroglobulinemia, rhabdomyosarcoma.

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