Pololike kinase 1 inhibitors

Abstract

This invention relates to inhibitors of polo-like kinase 1 (“PLK1”). Specifically, this invention relates to compounds that selectively inhibit PLK1 activity, pharmaceutical compositions comprising a therapeutically effective amount of the compound, and methods of using the compounds, such as methods for treating PLK1-mediated diseases and conditions such as cancer.

Description

Technical Field

[0001] This invention relates to inhibitors of polo-like kinase 1 (“PLK1”). Specifically, this invention relates to compounds that selectively inhibit PLK1 activity, pharmaceutical compositions comprising a therapeutically effective amount of the compound, and methods of using the compounds, such as methods for treating PLK1-mediated diseases and conditions such as cancer. Background Technology

[0002] TP53 It is the most frequently mutated and inactivated gene in cancer, leading to reduced levels of functional p53. Because existing approaches designed to directly target p53 have shown limited clinical success, a high unmet medical need remains for novel therapeutics to address cancers driven by p53 biological changes. Summary of the Invention

[0003] Compounds of formula (I) and their pharmaceutically acceptable salts are provided:

[0004] Formula (I) Where R 1 R 2 R 3 R 6 and R 7 As defined in this article.

[0005] Pharmaceutical compositions are also provided that comprise a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient or elixir.

[0006] A method is provided for treating PLK1-mediated diseases or conditions in subjects in need, the method comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutical salt thereof or a pharmaceutical composition thereof.

[0007] This document also provides for the use of compounds of formula (I) as defined herein, or pharmaceutically acceptable salts thereof, in the manufacture of pharmaceutical agents for inhibiting PLK1 activity.

[0008] This document also provides for the use of compounds of formula (I) as defined herein, or pharmaceutically acceptable salts or solvates thereof, in the manufacture of medicaments for the treatment of PLK1-mediated diseases or conditions. Detailed Implementation

[0009] This invention relates to inhibitors of polo-like kinase 1 (“PLK1”). Specifically, this invention relates to compounds that covalently inhibit PLK1 activity, pharmaceutical compositions comprising therapeutically effective amounts of the compounds, and methods of using these compositions, such as methods for treating plk1-mediated diseases and conditions such as cancer.

[0010] PLK1 Polo-like kinase 1 (PLK1), a member of the polo subfamily of serine / threonine protein kinases, is important for a variety of cellular functions, including the regulation of cell cycle progression and DNA damage response (DDR). Overexpression of PLK1, leading to increased cell proliferation due to disruption of cellular checkpoints, has been observed in a wide range of human cancers, solid tumors, and hematologic malignancies, and has been found to be associated with poorer clinical outcomes and resistance to chemotherapy. Conversely, PLK1 depletion or inhibition leads to mitotic arrest and DNA damage, often accompanied by cell death, and enhances the radiation sensitivity of cancer cells. The crucial role of PLK1 in cancer is widely accepted and considered unquestionable.

[0011] As a component of DDR, PLK1 plays a role in the oscillatory behavior of p53 activity. p53 is a regulatory tumor repressor protein important for DNA stability and is known as a guardian of the genome. PLK1 kinase activity promotes the proteasome degradation of p53 via the E3 ligases TOPORS and MDM2, where PLK1 depletion or inhibition leads to elevated cellular levels of p53. Furthermore, active PLK1 can physically bind to and inhibit p53 activity to transactivate the transcription of tumor repressor genes. Polo-like kinase 3 (PLK3), another member of the polo subfamily of kinases, has an opposite role to PLK1 in p53 activity; PLK3-mediated p53 phosphorylation increases its transactivation function.

[0012] While pan-PLK inhibitors can counteract p53 function, PLK1-specific inhibitors, namely the PLK1-specific inhibitors disclosed herein, can selectively increase p53 levels and activity. (Discovery) TP53 It mutates in about 50% of human cancers, more than any other gene, and TP53 Mutations in p53 are among the most common sudden changes in refractory cancers (see, for example, Hsiehchen et al., (2022) Nat. Comm. 13: 7477). Since no effective p53-based therapeutics have been successfully translated into clinical treatment, the PLK1-specific inhibitors disclosed herein may be able to generate therapeutic benefits by increasing p53 levels in cancer cells, including but not limited to those that are p53 deficient or p53 altered.

[0013] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. All patents, patent applications, and publications mentioned herein are incorporated by reference within the scope consistent with this disclosure. Unless otherwise expressly defined, terms and scope have their general definitions.

[0014] For simplicity, the chemical parts are primarily defined and referred to as monovalent chemical parts (e.g., alkyl, aryl, etc.). However, such terms may also be used to convey corresponding polyvalent parts where appropriate structures are clear to those skilled in the art. All atoms are understood to have their normal valence for bond formation (i.e., carbon is 4, N is 3, O is 2, and S is 2, 4, or 6 depending on its oxidation state).

[0015] As used in this article, "PLK1" refers to the mammalian enzyme polo-like kinase 1.

[0016] As used herein, "PLK1 inhibitor" refers to the compounds of the present invention represented by formula (I) as described herein. These compounds are capable of negatively regulating or inhibiting all or part of the enzymatic activity of PLK1 by forming a covalent adduct between the compound of formula (I) and PLK1.

[0017] As used in this article, the term "hydroxyl group" refers to -OH.

[0018] As used in this article, the term "hydroxyalkyl" refers to -alkylene-OH.

[0019] As used herein, the term "alkoxy" refers to -OC1-C3 alkyl.

[0020] As used in this article, the term "halogen" or "halogenated" refers to chlorine, bromine, fluorine, or iodine.

[0021] As used herein, the term "alkyl" refers to a straight-chain and branched aliphatic group having 1 to 12 carbon atoms. Therefore, "alkyl" encompasses C1, C2, C3, C4, C5, C6, C7, C8, C9, C1 ...1, C1, C1, C1, C1, C1, C1, C1 10 C 11 and C 12 Alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.

[0022] The term "alkylene group" refers to an alkyl group as defined above, which is located between and connects two other chemical groups. Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, and butylene.

[0023] As used herein, the term "alkynyl" refers to a straight-chain or branched aliphatic group having 2 to 6 carbon atoms and containing at least one carbon-carbon triple bond. Therefore, "alkynyl" encompasses C2, C3, C4, C5, and C6 groups. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentyynyl, and hexynyl.

[0024] The term "haloalkyl" refers to an alkyl chain in which one or more hydrogen atoms have been replaced by a halogen. Exemplary haloalkyl groups are trifluoromethyl, difluoromethyl, fluorochloromethyl, and fluoromethyl.

[0025] As used herein, "cycloalkyl" refers to saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 12 carbon atoms. Therefore, "cycloalkyl" includes C3, C4, C5, C6, C7, C8, C9, C16, C17, C18, C19, C18, C19, C19, C10, C11, C12, C13, C14, C15, C16, C17, C18, 10 C 11 and C 12 Cycloalkyl groups. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0026] An "aryl" group is a C6-C14 aromatic moiety comprising one to three aromatic rings. Therefore, "aryl" includes C6, C10, C13, and C14 cyclic hydrocarbon groups. Exemplary aryl groups are C6-C10 aryl groups. Specific aryl groups include, but are not limited to, phenyl, naphthyl, anthracene, and fluorenyl. "Aryl" groups also include fused polycyclic (e.g., bicyclic) ring systems, wherein one or more fused rings are non-aromatic rings, provided that at least one ring is an aromatic ring, such as an indene.

[0027] A "heterocyclic group" or "heterocyclic" group is a monocyclic or bicyclic (fused or spirocyclic) ring structure having 3 to 12 atoms (3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 atoms), for example, 4 to 8 atoms, wherein one or more ring atoms are independently -C(O)-, N, NR. A The ring must contain an O or S atom, and the remaining ring atoms must be quaternary carbons or carbonyl carbons. Examples of heterocyclic groups include, but are not limited to, epoxy, ethylene oxide, oxetane, aziridine, aziridinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrophenylthio, pyrrolyl, piperidinyl, piperazine, imidazoalkyl, thiazoalkyl, thiazoalkyl, dithiazoalkyl, trithiazoalkyl, azithienyl, oxathienyl, dioxopentyl, oxazolidinyl, oxazolidinone, decahydroquinolinyl, piperidinone, 4-piperidinone, 1,4-diazacycloheptane, 1-methyl-1,4-diazacycloheptane, 1,4-dimethyl-1,4-di(11-oxaneyl)-114,414-piperazinyl, thiomorpholinyl, dimethylmorpholinyl, and morpholinyl. Compounds having an ortho-ring O and / or S atom are specifically excluded from the scope of this terminology.

[0028] As used in this article, "L-heterocyclic group" refers to a heterocyclic group covalently linked to another group via an alkylene linker.

[0029] As used in this article, the “effective amount” of a compound is an amount sufficient to negatively regulate or inhibit PLK1 activity.

[0030] As used herein, a "therapeuticly effective amount" of a compound is an amount sufficient to improve or alleviate symptoms in some way, or to stop or reverse the progression of a condition (e.g., cancer), or to negatively regulate or inhibit PLK1 activity. Such amounts may be administered as a single dose or according to a prescribed regimen, thereby being effective.

[0031] As used in this article, “treatment” means any way that improves or otherwise beneficially alters a patient’s condition, symptoms, or pathology of a disease.

[0032] As used herein, improvement of symptoms of a particular condition by application of a particular compound or pharmaceutical composition means any relief attributable to or related to the application of the composition, whether permanent or temporary, continuous or transient.

[0033] compound In certain aspects of this disclosure, a compound of formula (I) and a pharmaceutically acceptable salt thereof are provided:

[0034] Formula (I) in: R 1 It can be hydrogen, halogen, hydroxyl, aniline, C1-C6 alkyl, cycloalkyl, and haloalkyl; R 2 It is a hydrogen, alkoxy, or -O-haloalkyl group optionally substituted with one or more alkoxy groups; R 3 Hydrogen, optionally with one or more R 4 Substituted heterocyclic groups, -NH-L 1 -N(R A R B -NH-L 2 -cycloalkyl or -NH-L 2 - Heterocyclic group, wherein the cycloalkyl group or the heterocyclic group is optionally surrounded by one or more R groups. 5 replace; R 6 It can be OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azacyclobutane, or S(O)(O)CH3; R 7It can be NH2, NHCH3, or CH3; Each R 4 Independently C1-C4 alkyl, cyclopropyl, hydroxyalkyl, -L 1 -N(R A R B ), halogen, O, O-CH3, NH2, -C(O)O-tert-butyl; -L 3 -aryl; or -L 4 - Heterocyclic group; Each R 5 Independently, it is a C1-C4 alkyl group, N(CH3)(CH3) or a heterocyclic group optionally substituted with an aryl group; Each L 1 It is a C1-C4 alkylene group; Each L 2 It is a bond or a C1-C4 alkylene group; Each L 3 It is -CH2-O-CH2-; Each L 4 For bond or methylene; Each R A It is hydrogen or C1-C3 alkyl; Each R B It is hydrogen or C1-C3 alkyl.

[0035] In some respects of the compounds of formula (I), R 1 The halogen is a halogen. In one embodiment, the halogen is chlorine, fluorine, or bromine.

[0036] In some respects of the compounds of formula (I), R 1 It is a hydroxyl group.

[0037] In some respects of the compounds of formula (I), R 1 It is a C1-C6 alkyl group. In one embodiment, the C1-C6 alkyl group is methyl, ethyl, or isopropyl.

[0038] In some respects of the compounds of formula (I), R 1 It is a cycloalkyl group. In one embodiment, the cycloalkyl group is cyclopropyl.

[0039] In some respects of the compounds of formula (I), R 1 The alkyl group is a haloalkyl group. In one embodiment, the haloalkyl group is fluoromethyl, difluoromethyl, or trifluoromethyl.

[0040] In some respects of the compounds of formula (I), R 1 The form is aniline. In one embodiment, aniline is NH2.

[0041] In some respects of the compounds of formula (I), R2 The alkoxy group is an alkoxy group. In one embodiment, the alkoxy group is a methoxy or propoxy group.

[0042] In some respects of the compounds of formula (I), R 2 It is an -O-haloalkyl group. In one embodiment, the haloalkyl group is a difluoromethyl group.

[0043] In some respects of the compounds of formula (I), R 3 It is optionally controlled by one or more R 4 Substituted heterocyclic groups.

[0044] In one embodiment, the heterocyclic group is 1,4-diazacycloheptyl, 1-methyl-1,4-diazacycloheptyl, or 1,4-dimethyl-1,4-di(11-oxoalkyl)-114,414-piperazinyl.

[0045] In another embodiment, the heterocyclic group is pyrrolidinyl, piperidinyl, or piperazineyl, each optionally surrounded by one or more R groups. 4 Replace, where R 4 C1-C4 alkyl, hydroxyalkyl, -L 1 -N(R A R B -C(O)O-tert-butyl, -L 3 -Aryl or -L 4 - Heterocyclic group.

[0046] In one implementation, the heterocyclic group is formed by an R 4 Substituted pyrroleyl and R 4 For L 4 - Heterocyclic group, where L 4 It is methylene, and the heterocyclic group is pyrrolidinyl.

[0047] In another embodiment, the heterocyclic group is surrounded by an R 4 Substituted piperidinyl and R 4 For L 4 - Heterocyclic group, where L 4 The heterocyclic group is a pyrrolidinyl group. In one embodiment, the heterocyclic group is piperazineyl, and R... 4 C1-C4 alkyl, hydroxyalkyl, -L 1 -N(R A R B -C(O)O-tert-butyl; or -L 3 -Aryl.

[0048] In one embodiment, the piperazine group is substituted with an R 4 Replace, and R 4It is a C1-C4 alkyl group, wherein the C1-C4 alkyl group is methyl or ethyl.

[0049] In one embodiment, the piperazine group is substituted with an R 4 Replace, and R 4 -L 1 -N(R A R B ).

[0050] In one embodiment, the piperazine group is substituted with an R 4 Replace, and R 4 It is -C(O)O-tert-butyl.

[0051] In another embodiment, the piperazine group is divided by two R 4 Group substitution, wherein one R4 group is a C1-C4 alkyl group and the second R4 group is a hydroxyalkyl group or -L 3 -Aryl.

[0052] In some respects of the compounds of formula (I), R 3 -NH-L 1 -N(R A R B ).

[0053] In one implementation, L 1 It is propylidene, and R A and R B Each is a methyl group.

[0054] In one implementation, L 1 It is propylidene, and R A and R B Each is an ethyl group.

[0055] In some respects of the compounds of formula (I), R 3 -NH-L 2 - Heterocyclic group, wherein the heterocyclic group is optionally surrounded by one or more R 5 replace.

[0056] In one implementation, L 2 It is a bond, and the heterocyclic group is optionally bound by one or more R 5 Substituted pyrrolidinyl groups.

[0057] In one embodiment, the pyrroleyl group is reacted with an R 5 Replace, where R 5 It is ethyl or isopropyl. In another embodiment, L 2 It is a C1-C4 alkylene group, and the heterocyclic group is pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl, each optionally surrounded by one or more R groups. 5replace.

[0058] In one implementation, L 2 It is ethylene or propylene.

[0059] In one embodiment, the heterocyclic group is optionally surrounded by one or more R groups. 5 Substituted piperazine group, wherein R 5 It is a methyl group.

[0060] In another implementation, L 2 It is a dimethyl vinyl group, and the heterocyclic group is pyrrolidinyl, piperidinyl, or morpholinyl.

[0061] In another implementation, L 2 It is a dimethyl vinyl group, and the heterocyclic group is optionally surrounded by one or more R groups. 5 Substituted piperazine group, wherein R 5 It is a methyl group.

[0062] In some respects, compounds of formula (I) are: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

[0063] Compounds of formula (I) and their pharmaceutically acceptable salts can be formulated into pharmaceutical compositions.

[0064] This invention also covers intermediate products used in the preparation of the compounds of this invention. These intermediate products are disclosed in the embodiments and examples of this invention.

[0065] The compounds disclosed herein include all pharmaceutically acceptable isotopically labeled compounds, wherein one or more atoms of the disclosed compounds are replaced by atoms having the same atomic number but a different atomic mass or mass number than those commonly found in nature. Examples of isotopes that can be incorporated into the compounds disclosed herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as... 2 H, 3 H, 11 C 13 C 14 C 13 N、 15 N、 15 O、 17 O、 31 P, 32 P, 35 S, 18 F, 36 Cl、 123 I and 125 I. Such radiolabeled compounds can be used to determine or measure the effectiveness of compounds, for example, by characterizing the binding affinity and kinetics to PLK1, as well as other modes of action. When using radioactive isotopes such as tritium... 3 H and carbon-14, i.e. 14 When labeled with C, some of the compounds disclosed herein can also be used to characterize drug distribution and in vivo tissue expression of PLK1.

[0066] When using heavier isotopes such as deuterium 2 When H-labeled, the compounds disclosed herein can exhibit therapeutic advantages, such as higher metabolic stability and a longer in vivo half-life, which in turn can reduce the requirements for therapeutic administration.

[0067] To better characterize substrate receptor occupancy or tissue expression in live samples statically or longitudinally, positron emission tomography (PET) isotopes such as... 11 C 14 F, 15 O and 13 Replacing the atoms of the compounds disclosed herein with N allows for positron emission tomography (PET) studies.

[0068] The isotopically labeled compounds disclosed herein can be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the relevant embodiments and schemes, using appropriate isotopically labeled reagents instead of previously used unlabeled reagents.

[0069] Some of the compounds disclosed herein can exist as stereoisomers. The compounds disclosed herein include all stereoisomers, including pure individual stereoisomer formulations and enriched formulations, as well as racemic mixtures of such stereoisomers and individual diastereomers and enantiomers that can be separated according to methods known to those skilled in the art. Furthermore, the compounds disclosed herein include all individual tautomeric states of the compounds and mixtures thereof.

[0070] Pharmaceutical Composition In another aspect, the present invention provides pharmaceutical compositions comprising a PLK1 inhibitor according to the invention and pharmaceutically acceptable carriers, excipients, or diluents. The compounds of the present invention can be formulated by any method well known in the art and can be prepared for administration via any route, including but not limited to parenteral, oral, sublingual, transdermal, topical, subcutaneous, intranasal, intratracheal, or rectal administration. In some embodiments, the compounds of the present invention are administered intravenously in a hospital setting. In some other embodiments, administration is preferably via an oral route.

[0071] The characteristics of the carrier will depend on the route of administration. As used herein, the term "pharmaceutically acceptable" means a non-toxic material that is compatible with biological systems such as cells, cell cultures, tissues, or organisms and does not interfere with the bioavailability of the active ingredient. Therefore, compositions according to the invention may contain, in addition to inhibitors, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. Preparation of pharmaceutically acceptable formulations is described, for example, in Remington's Pharmaceutical Sciences, 18th edition (edited by A. Gennaro, Mack Publishing Co., Easton, Pa., 1990).

[0072] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the compounds identified above and exhibits minimal or no undesirable toxicological effects. Examples of such salts include, but are not limited to, acid addition salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.) and salts formed with organic acids (such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, dihydroxynaphthyl acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid). Compounds may also be administered in pharmaceutically acceptable quaternary salt forms known to those skilled in the art, specifically including those of formula NR. + Z -The quaternary ammonium salt, wherein R is hydrogen, alkyl or benzyl, and Z is a counterion, including chloride, bromide, iodide, O-alkyl, toluenesulfonate, methanesulfonate, sulfonate, phosphate or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamate, mandelic acid, benzoate and diphenylacetate).

[0073] The active compound is included in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective amount to the patient without causing serious toxicity in the treated patient. For all the above conditions, the dosage range of the active compound is from about 0.01 mg / kg to 300 mg / kg based on the recipient's body weight, preferably from 0.1 mg / kg to 100 mg / kg daily, and more generally from 0.5 mg / kg to about 25 mg / kg daily.

[0074] Typical local dose ranges in suitable carriers will be 0.01%–3% wt / wt. The effective dose range of pharmaceutically acceptable derivatives can be calculated based on the weight of the parent compound to be delivered. If the derivative itself exhibits activity, the effective dose can be estimated using the weight of the derivative or by other means known to those skilled in the art, as described above.

[0075] Pharmaceutical compositions including compounds of the present invention can be used in the methods described herein.

[0076] How to use In another aspect, the present invention provides a method for inhibiting PLK1 activity in cells, comprising contacting cells for which PLK1 activity needs to be inhibited with a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the compound or a pharmaceutically acceptable salt thereof.

[0077] The compositions and methods provided herein are considered particularly suitable for inhibiting PLK1 activity in cells, including cells overexpressing PLK1. In one embodiment, cells requiring inhibition of PLK1 activity are contacted with an effective amount of the compound of formula (I) to negatively regulate the activity of the PLK1 kinase. In other embodiments, a pharmaceutically acceptable salt or pharmaceutical composition containing a therapeutically effective amount of the compound of formula (I) may be used. In some embodiments, contacting cells with an effective or therapeutically effective amount of the compound of formula (I) occurs in vivo. In some embodiments, contacting cells with an effective or therapeutically effective amount of the compound of formula (I) occurs in vitro. In one embodiment, the therapeutically effective amount of the compound of formula (I) is from about 0.01 mg / kg to 300 mg / kg per day. In one embodiment, the therapeutically effective amount of the compound of formula (I) is from about 0.1 mg / kg to 100 mg / kg per day.

[0078] In one implementation, methods are provided for treating a patient with a PLK1-mediated disease or condition, comprising administering to the patient, alone or in combination with a pharmaceutically acceptable carrier, excipient, or diluent, a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0079] In one implementation, the PLK1-mediated disease or condition is caused by the overexpression of PLK1. In one implementation, the PLK1-mediated disease or condition is cancer. In one implementation, PLK1-mediated cancer is... TP53 Or Ras-mutated cancers. In one implementation, PLK1-mediated cancers include melanoma, non-small cell lung cancer (NSCLC), head and neck cancer, esophageal cancer, pharyngeal cancer, breast cancer, liver cancer, endometrial cancer, colorectal cancer, ovarian cancer, pancreatic cancer, or prostate cancer.

[0080] These methods are designed to modulate PLK1 activity by negatively regulating it, particularly in cases where cells overexpress the PLK1 enzyme. In some implementations, this is used to treat certain PLK1-mediated diseases or conditions, such as cancer. Cells / patients may be exposed to single or multiple doses depending on the specific treatment regimen to induce the desired negative regulation of PLK1. For example, hematologic or solid tumors can be monitored using well-known methods, including blood draws, X-rays, or other imaging techniques such as CT or MRI scans, to assess the effectiveness of treatment, and the dosage can be adjusted accordingly by the attending physician.

[0081] The concentration and route of administration to patients may vary depending on the severity of the disease. Compounds, their pharmaceutically acceptable salts, and pharmaceutical compositions containing such compounds and salts may also be administered co-administered with other compounds as pre- or post-operative adjuvants, or in combination with other treatments such as surgical interventions. The degree of PLK1 inhibition in patients can be monitored using well-known assays, including those in Example A, to assess the effectiveness of treatment.

[0082] General reaction schemes and examples The compounds of the present invention can be prepared using the synthetic methods and reaction schemes described herein, or using other reagents and conventional methods well known to those skilled in the art, using commercially available reagents.

[0083] For example, the intermediates used to prepare the compounds of the present invention and the compounds of formula (I) can be prepared according to general reaction schemes I-III: General Reaction Scheme I

[0084] Compound (I) can be prepared according to general reaction scheme I, where R... 1 For hydrogen, halogen, hydroxyl, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R 2 It is hydrogen, alkoxy, or optionally conjugated by one or more R 4 Substituted -O-haloalkyl, and R 3 It is optionally controlled by one or more R 4 Substituted heterocyclic groups, -NH-L 1 -N(R A R B ) or -NH-L 2 - Heterocyclic group, wherein the heterocyclic group is optionally surrounded by one or more R 5 Substitution. Compound 3 is an example of formula (I), where R 1 and R 2 For H, and R 3 It is a heterocyclic group optionally substituted with one or more R4 groups, -NH-L1-N(R A R B ) or -NH-L2- heterocyclic group, wherein the heterocyclic group is optionally surrounded by one or more R 5 Replacement. For example, in S N Under Ar conditions, dihaloquinazoline 1 was treated with aniline 2 at high temperature to form anilinequinazoline 3, which was then subjected to Pd-catalyzed cross-coupling conditions, such as Heck conditions with vinyl sulfone 19, to provide anilinequinazoline styrene sulfone 4. Anilinequinazoline styrene sulfone 4 was separated by chromatography, i.e., reversed-phase HPLC, to obtain the desired compound 4 of formula (I).

[0085] General Reaction Scheme II :

[0086] Compound (I) can be prepared according to general reaction scheme II, wherein R... 1 It is hydrogen, halogen, hydroxyl, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, and R 2 It is hydrogen, alkoxy, or optionally conjugated by one or more R 4 Substituted -O-haloalkyl, R 3 It is optionally controlled by one or more R 4 Substituted heterocyclic groups, -NH-L 1 -N(R A R B ) or -NH-L 2 - Heterocyclic group, where L 1 L 2 R A and R B As defined in the specification, and the heterocyclic group is optionally composed of one or more R... 5 Substitution. Compound 3 is an example of formula (I), where R 1 It is hydrogen, halogen, hydroxyl, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, and R 2 It is hydrogen, alkoxy, or optionally conjugated by one or more R 4 Substituted -O-haloalkyl groups. The dihaloquinazoline 1 is subjected to S-type reactions, for example, those involving nitroaniline 17 and a weak acid, at high temperatures. N Ar conditions were applied, and the resulting nitroaniline quinazoline 18 was subjected to reducing conditions, such as Fe(III)Cl / HCl, to give aminoaniline quinazoline 20. Aminoaniline quinazoline 20 was subjected to Pd-catalyzed cross-coupling conditions, such as Heck conditions with vinyl sulfone 19, to give aniline quinazoline styryl sulfone 20. Styryl sulfone 20 was subjected to Buchwald cross-coupling conditions, such as those with a palladium catalyst and an aryl halide, or reductive amination conditions with an alkyl halide or aldehyde, such as those with sodium cyanoborohydride, to provide the desired product 4. The crude mixture of 4 was separated by chromatography, i.e., reversed-phase HPLC, to give the desired compound of formula (I), 4.

[0087] intermediate products The following intermediates can be used to prepare compounds of formula (I).

[0088] Intermediate product A

[0089] Step 1A solution of A-1, namely 4-bromo-2-fluoro-3-methoxybenzaldehyde (300 mg, 1.29 mmol, 1 equivalent) and guanidine carbonate (186 mg, 1.03 mmol, 0.8 equivalent) in DMA (3 mL) was stirred at 140 °C for 4 hours. H₂O (3 mL) was added to the mixture. The mixture was filtered, and the filter cake was dried under reduced pressure to obtain the residue. A-2, namely 7-bromo-8-methoxy-quinazolin-2-amine (0.24 g, 945 μmol, 73% yield), was given as an orange solid. 1 H NMR (400MHz, DMSO- d 6) δ ppm 9.13(s, 1H), 7.49 (d, J = 8.4Hz, 1H), 7.36 (d, J = 8.4Hz, 1H), 7.13 (s, 2H), 3.97 (s, 3H).

[0090] Step 2 A mixture of A-2 (7-bromo-8-methoxy-quinazolin-2-amine, 140 mg, 551 μmol, 1 equivalent), TBAC (184 mg, 661 μmol, 185 μL, 1.2 equivalent), and TMSCl (239 mg, 2.20 mmol, 280 μL, 4 equivalent) in DCM (1 mL) was stirred at 20 °C under N2. A solution of tert-butyl nitrite (171 mg, 1.65 mmol, 197 μL, 3 equivalent) in DMF (0.4 mL) was added to the mixture at 20 °C under N2. The mixture was stirred at 45 °C for 4 hours. The reaction mixture was quenched by adding H2O (3 mL) and extracted with DCM (2 mL × 3). The combined organic layers were washed with brine (2 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by preparative TLC (SiO2, DCM:ethyl acetate = 40:1). Intermediate A, 7-bromo-2-chloro-8-methoxy-quinazoline, was obtained as a yellow solid (60 mg, 219 μmol, 40% yield). 1 H NMR (400MHz, DMSO- d 6) δ ppm 9.65 (s, 1H), 8.01 (d, J = 8.8Hz, 1H), 7.93(d, J = 8.4Hz, 1H), 4.09 (s, 3H).

[0091] Intermediate product B

[0092] Step 1: At 0°C and under N2, B-1, i.e., (4-bromo-2-fluoro-6-methylphenyl)methanol BH3·THF (1 M, 10.7 mL, 2.5 equivalents), was added to a solution of 4-bromo-2-fluoro-6-methylbenzoic acid (1 g, 4.29 mmol, 1 equivalent) in 10 mL of THF. The mixture was stirred at 50°C for 2 hours. MeOH (2 mL) was added dropwise to the solution at 20°C and under N2. The mixture was concentrated under reduced pressure (3 batches) to obtain a residue. B-2, i.e., (4-bromo-2-fluoro-6-methylphenyl)methanol (2.8 g, crude), was obtained as a white solid. 1 H NMR (400MHz, MeOD) δ ppm 7.22 (s, 1H), 7.16-7.14 (m, 1H), 4.63 (d, J =2.0Hz, 2H), 2.43 (s, 3H).

[0093] Step 2: A mixture of B-2 (4-bromo-2-fluoro-6-methylphenyl)methanol (2.8 g, 12.78 mmol, 1 equivalent) and MnO2 (11.11 g, 127.82 mmol, 10 equivalent) in DCM (30 mL) was stirred at 40 °C for 8 hours. The reaction was filtered and concentrated under reduced pressure to obtain the residue. The residue (combined at the same scale) was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1:0 to 10:1). 3.9 g of B-3 (4-bromo-2-fluoro-6-methylbenzaldehyde) as a yellow oil was obtained.

[0094] Step 3: A mixture of B-3, namely 4-bromo-2-fluoro-6-methylbenzaldehyde (2 g, 9.22 mmol, 1 equivalent) and guanidine carbonate (1.33 g, 7.37 mmol, 0.8 equivalent), in DMA (20 mL) was stirred at 140 °C for 4 hours. H₂O (100 mL) was added to the reaction mixture. The mixture was filtered, and the filter cake was dried under reduced pressure to obtain a solid. B-4, namely 7-bromo-5-methyl-quinazoline-2-amine (1.5 g, crude), was obtained as a brown solid. 1 H NMR (400MHz, DMSO- d 6) δ ppm 9.22 (s, 1H), 7.43 (s, 1H), 7.17 (s, 1H), 7.00 (s, 2H), 2.59 (s, 3H).

[0095] Step 4A mixture of B-4 (7-bromo-5-methylquinazolin-2-amine) (0.5 g, 2.10 mmol, 1 equivalent), TBAC (700.4 mg, 2.52 mmol, 705 μL, 1.2 equivalent), and TMSCl (913 mg, 8.40 mmol, 1.07 mL, 4 equivalent) in DCM (3 mL) was stirred at 20 °C under N2. A solution of tert-butyl nitrite (650 mg, 6.30 mmol, 749 μL, 3 equivalent) in DMF (2 mL) was added to the mixture at 20 °C under N2. The mixture was stirred at 45 °C for 4 hours. The reaction mixture (two batches) was quenched by adding H2O (20 mL) and extracted with DCM (20 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, DCM:ethyl acetate = 1:0 to 50:1). Intermediate 4, 7-bromo-2-chloro-5-methylquinazoline, was obtained as a yellow solid (0.100 g, 388 μmol, 10% yield). 1 H NMR (400MHz, DMSO- d 6) δ ppm 9.70 (s, 1H), 8.09 (s, 1H), 7.82 (s, 1H), 2.77 (s, 3H).

[0096] intermediate product C

[0097] Step 1: BH3·THF (1M, 10.7mL, 2.5 equivalents) was added to a solution of C-1, 4-bromo-2-fluoro-6-methylbenzoic acid (1g, 4.29mmol, 1 equivalent), in THF (10mL) at 0°C under N2. The mixture was stirred at 50°C for 2 hours. MeOH (2mL) was added dropwise to the solution at 20°C under N2. The mixture was concentrated under reduced pressure (3 batches) to obtain a residue. C-2, (4-bromo-2-fluoro-6-methylphenyl)methanol (2.8g, crude), was obtained as a white solid. 1 H NMR (400MHz, MeOD) δppm 7.22 (s, 1H), 7.16-7.14 (m, 1H), 4.63 (d, J = 2.0Hz, 2H), 2.43 (s, 3H).

[0098] Step 2: A mixture of C-2 (4-bromo-2-fluoro-6-methylphenyl)methanol (2.8 g, 12.78 mmol, 1 equivalent) and MnO2 (11.11 g, 127.82 mmol, 10 equivalent) in DCM (30 mL) was stirred at 40 °C for 8 hours. The reaction was filtered and concentrated under reduced pressure to obtain the residue. The residue (combined at the same scale) was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1:0 to 10:1). 3.9 g of C-3 (4-bromo-2-fluoro-6-methylbenzaldehyde) as a yellow oil was obtained.

[0099] Step 3: A mixture of C-3, 4-bromo-2-fluoro-6-methylbenzaldehyde (2 g, 9.22 mmol, 1 equivalent) and guanidine carbonate (1.33 g, 7.37 mmol, 0.8 equivalent), in DMA (20 mL) was stirred at 140 °C for 4 hours. H₂O (100 mL) was added to the reaction mixture. The mixture was filtered, and the filter cake was dried under reduced pressure to obtain the residue. C-4, 7-bromo-5-methyl-quinazolin-2-amine (1.5 g, crude), was obtained as a brown solid. ¹H NMR (400 MHz, DMSO-) d 6) δ ppm 9.22 (s, 1H), 7.43 (s, 1H), 7.17 (s, 1H), 7.00 (s, 2H), 2.59 (s, 3H).

[0100] Step 4: A mixture of C-4 (7-bromo-5-methylquinazolin-2-amine) (0.5 g, 2.10 mmol, 1 equivalent), TBAC (700.4 mg, 2.52 mmol, 705 μL, 1.2 equivalent), and TMSCl (913 mg, 8.40 mmol, 1.07 mL, 4 equivalent) in DCM (3 mL) was stirred at 20 °C under N2. A solution of tert-butyl nitrite (650.0 mg, 6.30 mmol, 749 μL, 3 equivalent) in DMF (2 mL) was added to the mixture at 20 °C under N2. The mixture was stirred at 45 °C for 4 hours. The reaction mixture (two batches) was quenched by adding H2O (20 mL) and extracted with DCM (20 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM:ethyl acetate = 1:0 to 50:1). Intermediate C, 7-bromo-2-chloro-5-methylquinazoline, was obtained as a yellow solid (0.1 g, 388 μmol, 10% yield). 1 H NMR (400MHz, DMSO- d6) δ ppm 9.70 (s, 1H), 8.09 (s, 1H), 7.82 (s, 1H), 2.77 (s, 3H).

[0101] Intermediate product D

[0102] Step 1: Guanidine carbonate (243 mg, 1.35 mmol, 0.8 equivalents) was added to a solution of D-1, namely 4-bromo-2-chloro-6-fluorobenzaldehyde (400 mg, 1.68 mmol, 1 equivalent), in DMA (4 mL). The mixture was stirred at 140 °C for 4 hours. The residue was diluted with H2O (20 mL), filtered, and the filter cake was concentrated under reduced pressure to obtain the residue. The product was used in the next step without further purification. D-2, namely 7-bromo-5-chloroquinazoline-2(1H)-imine (350 mg, 1.35 mmol, 80% yield), was obtained as a brown solid. 1 H NMR (400MHz, DMSO- d 6) δ = 9.25 (s, 1H), 7.59 (d, J =1.4Hz, 1H), 7.53 (d, J = 1.8Hz, 1H), 7.34 (s, 2H).

[0103] Step 2: To a solution of D-2 (7-bromo-5-chloro-quinazolin-2-amine, 350 mg, 1.35 mmol, 1 equivalent), TMSCl (588 mg, 5.42 mmol, 687 μL, 4 equivalents), and tetrabutylammonium chloride (452 ​​mg, 1.62 mmol, 454 μL, 1.2 equivalents) in DCM (2 mL), tert-butyl nitrite (418.86 mg, 4.06 mmol, 483 μL, 3 equivalents) in DMF (2 mL) was added. The mixture was stirred at 45 °C for 1 hour. The residue was diluted with H2O (10 mL) and extracted with DCM (10 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 1:0 to 10:1). Intermediate product D, 7-bromo-2,5-dichloro-quinazoline, was obtained as a white solid (160 mg, 576 μmol, 43% yield).

[0104] Intermediate product E

[0105] Step 1: A solution of E1 (10 g, 45.25 mmol, 1 equivalent), carbonic acid, and guanidine (6.52 g, 36.20 mmol, 0.8 equivalent) in DMA (300 mL) was stirred at 140 °C for 4 hours. Water (1000 mL) was added, and the mixture was stirred at 20 °C for 0.5 hours. After this time, a yellow solid precipitated. The solid was collected, filtered, and washed with water (1000 mL). The filter cake was dried under reduced pressure. The product was used directly in the next step without further purification. The compound 7-bromo-5-fluoro-quinazoline-2-amine (50 g, 196 mmol, 87% yield, 95% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ = 9.23(s, 1H), 7.44 (d, J = 0.8Hz, 1H), 7.32 (s, 2H), 7.26 (dd, J = 1.6, 9.3Hz, 1H) Step 2: CuI (7.9 g, 41.3 mmol, 1 equivalent), diiodomethane (55.3 g, 206.6 mmol, 17.00 mL, 5 equivalent), and amyl nitrite (14.5 g, 124.0 mmol, 17 mL, 3 equivalent) were added to a solution of E2 (10 g, 41.3 mmol, 1 equivalent) in THF (300 mL). The mixture was stirred at 80 °C for 2 hours. The mixture was slowly added to ice water (100 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous Na2SO4, filtered, and concentrated to give the residue. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate = 10:1 to 3:1). Intermediate E, 7-bromo-5-fluoro-2-iodo-quinazoline (6 g, 16.15 mmol, 13% yield, 95% purity), was given as a white solid. 1 H NMR (400MHz, DMSO-) d 6) δ = 9.45 (s, 1H), 8.11 (s, 1H), 7.96 (dd, J = 1.5, 9.1Hz, 1H) intermediate product F

[0106] Step 1: NaOtBu (2M, 4.50 mL, 1 equivalent) and Davephos Pd G3 (34.36 mg, 45.02 μmol, 0.01 equivalent) were added to a solution of 5-nitro-2-(trifluoromethoxy)aniline (1 g, 4.50 mL, 1 equivalent) in DMF (30 mL). The mixture was stirred at 130 °C for 12 hours. Water (200 mL) was added, and the aqueous layer was extracted with DCM (50 mL × 3). The combined organic phases were dried over anhydrous Na₂SO₄, filtered, and concentrated to obtain the residue. The product was used in the next step without purification. 7-Bromo-5-fluoro-N-[5-nitro-2-(trifluoromethoxy)phenyl]quinazoline-2-amine (4 g, crude) was obtained as a yellow solid.

[0107] Step 2: A solution of 7-bromo-5-fluoro-N-[5-nitro-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (4 g, 8.95 mmol, 1 equivalent) in THF (30 mL), H₂O (10 mL), and Fe (4.00 g, 71.57 mmol, 8 equivalent) and NH₄Cl (3.83 g, 71.57 mmol, 8 equivalent) was stirred at 60 °C for 12 hours. The mixture was filtered and the filtrate was concentrated to obtain the residue. The residue was partitioned between water (200 mL) and DCM (200 mL), and the aqueous layer was extracted with DCM (3 × 100 mL). The combined organic phases were dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate = 2:1 to 0:1). The intermediate product F, namely N3-(7-bromo-5-fluoro-quinazolin-2-yl)-4-(trifluoromethoxy)benzene-1,3-diamine (1.4 g, 3.19 mmol, 36% yield, 95% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ =9.39 (s, 1H), 9.30 (s, 1H), 7.65 (s, 1H), 7.45 (br d, J = 8.3Hz, 1H), 7.23(br d, J = 2.0Hz, 1H), 7.04 (br d, J = 8.5Hz, 1H), 6.41 (dd, J = 2.6, 8.8Hz, 1H), 5.36 (s, 2H).

[0108] intermediate product G

[0109] Step 1: 4-Bromo-2,6-difluorobenzaldehyde (20 g, 90.50 mmol, 1 equivalent) and guanidine carbonate (13.04 g, 72.40 mmol, 0.8 equivalent) were added to DMA (250 mL). The mixture was stirred at 140 °C for 4 hours. Water (1000 mL) was added to the mixture, and the solid was allowed to precipitate. The crude product (combined with other batches in the same scale) was filtered and washed with water (1000 mL). The solid was collected and dried under reduced pressure. The product was used in the next step without further purification. The compound 7-bromo-5-fluoro-quinazoline-2-amine (57 g, 200.17 mmol, 74% yield, 85% purity) was given as a yellow solid. 1 H NMR (400MHz, DMSO-) d 6) δ = 9.23 (s, 1H), 7.44 (s, 1H), 7.32 (s, 2H), 7.27 (d, J = 9.6Hz, 1H).

[0110] Step 2: Add pyr dropwise to a mixture of 7-bromo-5-fluoro-quinazolin-2-amine (10 g, 41.31 mmol, 1 equivalent) in DCM (30 mL) at 0 °C. HF (20 mL). Then t-BuONO (12.78 g, 124 mmol, 15 mL, 3 equivalents) was added dropwise to the mixture at 0 °C. The mixture was stirred at 0 °C for 2 hours. The reaction mixture was poured into ice water (50 mL) and extracted with EtOAc (20 mL × 3). The combined organic phases were washed with saturated NaHCO3 (20 mL) and brine (20 mL), dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 100:1 to 50:1). The compound 7-bromo-2,5-difluoro-quinazoline (17.5 g, 71.42 mmol, 35% yield) was given as a white solid. 1 H NMR (400MHz, CDCl3) -d ) δ = 9.61 (s, 1H), 8.01 (s, 1H), 7.48 (d, J = 8.8Hz, 1H).

[0111] intermediate product H

[0112] Step 1The mixture of N-3-(7-bromo-5-fluoro-quinazolin-2-yl)-4-(trifluoromethoxy)phenyl-1,3-diamine (3 g, 7.2 mmol, 1 equivalent), ethylene sulfonamide (925 mg, 8.63 mmol, 1.2 equivalent), Pd(OAc)2 (32.3 mg, 144 μmol, 0.02 equivalent), tri-o-tolylphosphine (88 mg, 287.66 μmol, 0.04 equivalent), and TEA (2.18 g, 21.57 mmol, 3.00 mL, 3 equivalent) in DMF (30 mL) was degassed and purged three times with N2. The mixture was then stirred at 100 °C for 12 hours under N2 atmosphere. The reaction mixture was filtered and concentrated to remove the solvent. The crude product was milled at 20 °C for 30 minutes with PE:EtOAc (5:1, 20 mL). The compound (E)-2-[2-[5-amino-2-(trifluoromethoxy)anilino]-5-fluoro-quinazolin-7-yl]ethylenesulfonamide (2.7 g, 6.09 mmol, 85% yield) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ = 9.38 (s, 1H), 9.23 (s, 1H), 7.65 (s, 1H), 7.61 (d, J = 11.2Hz, 1H),7.57-7.49 (m, 1H), 7.46-7.37 (m, 1H), 7.25 (br s, 2H), 7.14 (d, J = 2.8Hz,1H), 7.05-7.01 (m, 1H), 6.43-6.38 (m, 1H), 5.33 (s, 2H).

[0113] Step 2At 0 °C, a solution of (E)-2-[2-[5-amino-2-(trifluoromethoxy)anilino]-5-fluoro-quinazolin-7-yl]ethylenesulfonamide (500 mg, 1.13 mmol, 1 equivalent) and p-TsOH·H2O (643.53 mg, 3.38 mmol, 3 equivalents) in ACN (12 mL) was added dropwise to a solution of NaNO2 (155.61 mg, 2.26 mmol, 2 equivalents) and KI (468.01 mg, 2.82 mmol, 2.5 equivalents) in H2O (4 mL). The mixture was stirred at 20 °C under N2 for 2 hours. The reaction mixture was quenched by adding H2O (60 mL) and an aqueous solution of Na2S2O3 (2 M, 6 mL), followed by extraction with EtOAc (10 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The crude product was ground with EtOAc (10 mL) at 20 °C for 20 minutes. (E)-2-[5-fluoro-2-[5-iodo-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (400 mg, 722 μmol, 64% yield) was obtained as a pink solid. 1 H NMR (400MHz, DMSO- d 6) δ = 9.74 (s, 1H), 9.44 (s, 1H), 8.36 (d, J = 2.0Hz, 1H), 7.74-7.64 (m, 2H), 7.64-7.59 (m, 1H), 7.55 (s,1H), 7.51-7.44 (m, 1H), 7.28-7.21 (m, 3H).

[0114] Intermediate product I-001

[0115] Step 1A mixture of 5-bromo-2-(trifluoromethoxy)aniline (50 mg, 195.30 μmol, 1 equivalent), 1-methylpiperazine (23.47 mg, 234.36 μmol, 26.00 μL, 1.2 equivalent), NaOBu-t (37.54 mg, 390.59 μmol, 2 equivalent), and [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium; di-tert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphonane (15.51 mg, 19.53 μmol, 0.1 equivalent) in dioxane (5 mL) was degassed and purged three times with N2. The mixture was then stirred at 90 °C for 12 hours under N2 atmosphere. The mixture was filtered through diatomaceous earth, and the solvent was removed under reduced pressure to provide the residue. The residue was purified by preparative HPLC (column: Waters Xbridge BEH C18 250×50mm×10μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 25%-50%, 10 min). 180 mg of the compound 5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)aniline, a white solid, was obtained, i.e., intermediate-001. 1 H NMR (400MHz, DMSO- d 6) δ = 6.89 (d, J = 8.8Hz, 1H), 6.32 (d, J = 2.8Hz, 1H), 6.14 (dd, J = 2.8Hz, 8.8Hz,1H), 5.11 (br s, 2H), 3.04 - 3.02 (m, 4H), 2.42-2.40 (m, 4H), 2.20 (s, 3H).

[0116] Intermediates I-002 to I-036 shown in Table A were prepared according to the teachings of the general reaction scheme and the method for preparing intermediate I-001.

[0117] Table A

[0118]

[0119]

[0120]

[0121] Intermediate product I-037

[0122] Step 1 A solution of NaSMe (1.05 g, 14.98 mmol, 954.55 μL, 1.10 equivalent) in H2O (25 mL) was added dropwise to a solution of 4-bromo-1-fluoro-2-nitrobenzene I-037-1 (3 g, 13.64 mmol, 1.68 mL, 1 equivalent) in DMF (75 mL) at 0 °C under N2. The mixture was stirred at 0 °C under N2 for 1 hour. The mixture was filtered to obtain a filter cake and washed with H2O (10 mL). Brine (20 mL) was added to the residual filtrate, and then extracted with EtOAc (10 mL × 3). The combined organic layers were dried over Na2SO4. Na2SO4 was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 50 / 1). The compound 4-bromo-1-methylthio-2-nitrobenzene (3 g, 12.09 mmol, 89% yield) was given as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ = 8.37 (d, J =2.2Hz, 1H), 7.91-7.95 (m, 1H), 7.53 (d, J = 8.6Hz, 1H), 2.54 (s, 3H).

[0123] Step 2: A mixture of 4-bromo-1-methylthio-2-nitrobenzene (2 g, 8.06 mmol, 1 equivalent), 1-methylpiperazine (1.61 g, 16.12 mmol, 1.79 mL, 2 equivalents), Cs₂CO₃ (5.25 g, 16.12 mmol, 2 equivalents), Xantphos (466.45 mg, 806.14 μmol, 0.1 equivalents), and Pd₂(dba)₃ (369.10 mg, 403.07 μmol, 0.05 equivalents) in dioxane (30 mL) was degassed and purged three times with N₂. The mixture was then stirred at 90 °C for 12 hours under N₂ atmosphere. The filtrate was filtered and concentrated to remove the solvent. The residue was purified by preparative HPLC (FA conditions; column: Phenomenex luna C18 (250 × 70 mm, 10 μm); mobile phase: [water (FA)-ACN]; gradient: 5%–30% B, for 25 min). The compound 1-methyl-4-(4-methylthio-3-nitro-phenyl)piperazine (1.5 g, 5.61 mmol, 70% yield) was obtained as a brown oil. 1 HNMR (400MHz, CDCl3) δ = 7.73 (d, J= 2.6Hz, 1H), 7.24-7.27 (m, 1H), 7.14-7.17(m, 1H), 3.39 (br t, J = 4.6Hz, 4H), 3.03 (br t, J = 4.6Hz, 4H), 2.61 (s, 3H), 2.45 (s, 3H).

[0124] Step 3 Fe (522.22 mg, 9.35 mmol, 5 equivalents) was added to a solution of 1-methyl-4-(4-methylthio-3-nitro-phenyl)piperazine (500 mg, 1.87 mmol, 1 equivalent) and NH4Cl (500.21 mg, 9.35 mmol, 5 equivalents) in EtOH (6 mL) and H2O (3 mL) at 50 °C. After addition, the resulting mixture was stirred at 85 °C for 2 hours. The mixture was filtered to obtain a filtrate, which was then concentrated to remove the solvent. No further purification was required, and it was used directly in the next step. The compound 5-(4-methylpiperazin-1-yl)-2-methylthioaniline (220 mg, crude) was obtained as a brown solid. m / z (ES + ) [M+H]+= 238.1.

[0125] Intermediates I-038 to I-045 shown in Table B were prepared according to the teachings of the general reaction scheme and the method for preparing intermediate I-037.

[0126] Table B

[0127] Intermediate product I-046

[0128] Step 1KHMDS (1M, 15.62mL, 2 equivalents) was added dropwise to a solution of 5-bromo-2-(trifluoromethoxy)aniline (2g, 7.81mmol, 1 equivalent) in THF (20mL) at 0°C and stirred for 0.5 hours. Then, Boc₂O (1.70g, 7.81mmol, 1.79mL, 1 equivalent) was added fractionally at 0°C. The mixture was stirred at 25°C for 2 hours. The mixture was quenched with saturated NH₄Cl (50mL) and extracted with ethyl acetate (20mL × 3). The combined organic phases were dried over anhydrous Na₂SO₄, filtered, and concentrated to give the residue. The crude product was ground with petroleum ether / ethyl acetate (5:1, 10 mL) at 15 °C for 10 minutes, filtered, and the filter cake was dried under vacuum to give N-[5-bromo-2-(trifluoromethoxy)phenyl]carbamate tert-butyl ester (1.5 g, 4.21 mmol, 54% yield) as a white solid. 1 H NMR (400MHz, DMSO- d 6) δ = 9.33 (s, 1H), 7.81 (d, J =2.0Hz, 1H), 7.42- 7.38 (dd, 1H), 7.37-7.33 (dd, 1H), 1.50 (s, 9H).

[0129] Step 2 Pd(dppf)Cl2 (287.64 mg, 393.11 μmol, 0.1 equivalent) and KOAc (1.16 g, 11.79 mmol, 3.5 equivalent) were added to a solution of N-[5-bromo-2-(trifluoromethoxy)phenyl]carbamate tert-butyl ester (1.4 g, 3.93 mmol, 1 equivalent) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1,3,2-dioxaborhexacyclopentaborane (2.00 g, 7.86 mmol, 2 equivalent) in dioxane (10 mL). The mixture was stirred at 100 °C for 12 hours under N2. Water (50 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phases were dried over anhydrous Na₂SO₄, filtered, and concentrated to obtain the residue. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 100 / 1, 50 / 1). The compound N-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-2-(trifluoromethoxy)phenyl]carbamate tert-butyl ester (1.2 g, 2.98 mmol, 76% yield) was obtained as a white oil. 1 H NMR (400MHz, DMSO- d6) δ = 9.02 (s, 1H), 7.97 (s, 1H), 7.47 (d, J =8.2Hz, 1H), 7.34 (d, J = 8.2Hz, 1H), 1.46 (s, 9H), 1.29 (s, 12H).

[0130] Step 3 Pd(dppf)Cl2 (18.15 mg, 24.80 μmol, 0.01 equivalent) and K2CO3 (1.03 g, 7.44 mmol, 3 equivalent) were added to a solution of N-[5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)-2-(trifluoromethoxy)phenyl]carbamate (1 g, 2.48 mmol, 1 equivalent) and 3-bromo-1H-pyridazin-6-one (520.78 mg, 2.98 mmol, 1.2 equivalent) in dioxane (40 mL) and H2O (4 mL). The mixture was stirred at 100 °C for 12 hours. Water (50 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (20 mL × 3). The combined organic phases were dried over anhydrous Na2SO4, filtered, and concentrated to give the residue. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 100 / 1, 50 / 1). 600 mg of the compound N-[5-(6-oxo-1H-pyridazin-3-yl)-2-(trifluoromethoxy)phenyl]carbamate tert-butyl ester was obtained as a white oil. 1 H NMR (400MHz, DMSO- d 6) δ = 13.27 (s, 1H), 9.17 (br s,1H), 8.22 (br s, 1H), 8.00 (br dd, J = 2.8, 9.7Hz, 1H), 7.71 - 7.62 (m, 1H),7.45 (d, J = 6.7Hz, 1H), 7.01 (br dd, J = 3.0, 9.8Hz, 1H), 1.47 (s, 9H).

[0131] Step 4A mixture of N-[5-(6-oxo-1H-pyridazin-3-yl)-2-(trifluoromethoxy)phenyl]carbamate tert-butyl ester (600 mg, 1.62 mmol, 1 equivalent) in HCl / EtOAc (4 M, 5 mL, 12.38 equivalent) was stirred at 20 °C for 1 hour. The mixture was concentrated under vacuum. The product was used in the next step without purification. The compound 3-[3-amino-4-(trifluoromethoxy)phenyl]-1H-pyridazin-6-one (350 mg, 1.14 mmol, 70% yield, HCl salt) was given as a white solid. 1 H NMR (400MHz, DMSO-) d 6) δ = 13.20 (s, 1H), 7.89 (d, J = 9.9Hz, 1H), 7.35 (d, J =2.1Hz, 1H), 7.20 (d, J = 8.5Hz, 1H), 7.05 (dd, J = 2.1, 8.5Hz, 1H), 6.97 (d,J = 9.9Hz, 1H) Intermediates I-047 to I-064 shown in Table C were prepared according to the teachings of the general reaction scheme and the method for preparing intermediate I-046.

[0132] Table C

[0133]

[0134]

[0135] Example The following examples are intended to further illustrate certain embodiments of the present invention, and are not intended to limit the scope of the invention.

[0136] Example 1

[0137] Step 14-Methylbenzenesulfonic acid hydrate (415 mg, 2.18 mmol, 1.2 equivalents) was added in a single step to a mixture of 7-bromo-2-chloroquinazoline (663 mg, 2.72 mmol, 1.5 equivalents) and 5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)aniline (500 mg, 1.82 mmol, 1 equivalent) in DMF (6 mL) under N2 conditions. The mixture was stirred at 130 °C for 12 h. H2O (60 mL) was added to the mixture and it was extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 0 / 1 to ethyl acetate / MeOH = 3 / 1). The compound 7-bromo-N-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (500 mg, 1.04 mmol, 57% yield) was obtained as a white solid.

[0138] Step 2 Under N2, TEA (210 mg, 2.07 μmol, 1 equivalent), and Pd(OAc)2 (4.66 mg, 20.7 μmol, 0.1 equivalent) were added in a single batch to a mixture of 7-bromo-N-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (100 mg, 207 μmol, 1 equivalent), tri-o-tolylphosphine (25 mg, 83.0 μmol, 0.4 equivalent), and ethylene sulfonamide (67 mg, 622 μmol, 15.0 μL, 3 equivalent) in DMF (0.3 mL). The mixture was stirred at 80 °C for 12 hours. The mixtures were combined in 15 mg increments. The mixture was filtered to obtain a filtrate, which was then concentrated under vacuum. Preparative HPLC purification of the residue (column: Waters XBridge BEH C18 100×30mm×10μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 35%-65%, 8 min). (E)-2-[2-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (10 mg, 0.2 mmol, 10% yield, 100% purity) was obtained as a white solid. 1 H NMR (400MHz, CD3CN) δ ppm 9.20 (s, 1H), 8.37 (d, J = 2.8Hz, 1H), 7.90 (d, J = 8.4Hz, 1H),7.86 (s, 1H), 7.71 (s, 1H), 7.64-7.59 (m, 2H), 7.29 (d,J = 8.4Hz, 1H), 7.24-7.21 (m, 1H), 6.70-6.67 (m, 1H), 5.58 (s, 2H), 3.27 (t, J = 4.8Hz, 4H), 2.56(t, J = 4.8Hz, 4H), 2.15 (s, 3H). m / z (ES + [M+1] + = 509.1.

[0139] Example 2

[0140] Step 5 A solution of 7-bromo-2-chloro-5-methylquinazoline (0.1 g, 388 μmol, 1 equivalent), 5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)aniline (107 mg, 388 μmol, 1 equivalent), and TFA (89 mg, 777 μmol, 58 μL, 2 equivalent) in n-BuOH (10 mL) was stirred at 90 °C for 12 h. The mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (neutral conditions; column: Waters XBridge Prep OBD C18 150 × 40 mm × 10 μm; mobile phase: [H2O (10 mM NH4HCO3)-ACN]; gradient: 45%-75% B, for 8.0 min). 7-Bromo-5-methyl-N-(5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)quinazolin-2-amine 2F was obtained as a white solid (40 mg, 81 μmol, 21% yield). 1 H NMR (400MHz, DMSO- d 6) δ ppm 9.40 (s, 1H), 9.20 (s, 1H), 7.60 (d, J =2.8Hz, 2H), 7.36 (s, 1H), 7.21 (d, J = 8.8Hz, 1H), 6.80-6.77 (m, 1H), 3.19-3.17 (m, 4H), 2.66 (s, 3H), 2.48-2.46 (m, 4H), 2.23 (s, 3H).

[0141] Step 6Under N2 conditions, Tris-o-tolylphosphine (2.45 mg, 8.1 μmol, 0.1 equivalent) and Pd(OAc)2 (1.81 mg, 8.06 μmol, 0.1 equivalent) were added to a solution of 7-bromo-5-methyl-N-(5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (40 mg, 81.0 μmol, 1 equivalent), ethylene sulfonamide (8.6 mg, 81 μmol, 1 equivalent), and TEA (16 mg, 161 μmol, 22.4 μL, 2 equivalent) in DMF (1 mL). The mixture was stirred at 100 °C for 12 hours. The mixture (combined in 60 mg increments) was concentrated under vacuum without further post-treatment. The residue was subjected to preparative HPLC (neutral conditions; column: Waters XBridge Prep OBD C18). 150×40mm×10μm; mobile phase: [H2O (10mMNH4HCO3)-ACN]; gradient: 30%-60% B (for 8.0 min) purification. 25 mg of (E)-2-[5-methyl-2-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (Example 2, 98% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ ppm 9.41 (s, 1H), 9.10 (s, 1H), 7.69 (d, J =2.8Hz, 1H), 7.63 (s, 1H), 7.57 (m, 1H), 7.44 (d, J = 4.0Hz, 2H), 7.22-7.20 (m,3H), 6.79-6.76 (m, 1H), 3.20-3.18 (m, 4H), 2.68-2.66 (m, 3H), 2.49-2.46 (m,4H), 2.23 (s, 3H). m / z (ES + [M+1] + = 523.2.

[0142] Example 3

[0143] Step 1TFA (131.28 mg, 1.15 mmol, 86 μL, 2 equivalents) was added to a solution of 7-bromo-2,5-dichloroquinazoline (160 mg, 576 μmol, 1 equivalent) and 5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)aniline (158 mg, 576 μmol, 1 equivalent) in n-BuOH (10 mL). The mixture was stirred at 90 °C for 12 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge BEH C18 100 × 30 mm 5 µm; mobile phase: [H2O(10 mM NH4HCO3)-MeCN]; gradient: 55%-85% B, 8.0 min). The compound 7-bromo-5-chloro-N-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (60 mg, 116 μmol, 20% yield) was obtained as a yellow solid. m / z (ES + [M+1] + = 516.1.

[0144] Step 2 A mixture of 7-bromo-5-chloro-N-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (50 mg, 97 μmol, 1 equivalent), ethylene sulfonamide (12 mg, 116 μmol, 1.2 equivalent), Pd(OAc)2 (2.2 mg, 9.7 μmol, 0.1 equivalent), tri-o-tolylphosphine (3.0 mg, 10.0 μmol, 0.1 equivalent), and TEA (29 mg, 290 μmol, 40 μL, 3 equivalent) in DMF (1 mL) was degassed and purged three times with N2. The mixture was then stirred at 80 °C for 12 hours under N2 atmosphere. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge Prep OBDC18 150×40mm×10µm; mobile phase: [H2O(10mM NH4HCO3)-MeCN]; gradient: 35%-65% B, for 8.0 min). The compound (E)-2-[5-chloro-2-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (12 mg, 20 μmol, 21% yield, 91% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-) d6) δ = 9.41 (s, 1H), 7.94 (s, 1H), 7.79 (s, 1H), 7.65 - 7.59 (m, 1H), 7.52 (br d, J = 2.6Hz, 1H), 7.45 (d, J = 15.6Hz, 1H), 7.28 - 7.18 (m, 2H), 6.82 (dd, J = 2.8, 9.1Hz, 1H), 3.21 - 3.14 (m, 4H), 2.45 (br s, 4H), 2.22 (s, 3H). m / z (ES + [M+1] + = 543.1.

[0145] Example 4

[0146] Step 1 TFA (70 mg, 612 μmol, 46 μL, 2 equivalents) and 5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)aniline (84.22 mg, 305.95 μmol, 1 equivalent) were added to a solution of 7-bromo-2-chloro-5-fluoroquinazoline (80 mg, 306 μmol, 1 equivalent) in n-BuOH (8 mL). The mixture was stirred at 90 °C for 12 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: 2-Phenomenex Gemini C18 75 × 40 mm × 3 µm; mobile phase: [H2O(10 mMNH4HCO3)-MeCN]; gradient: 40%-75% B, 8.0 min). The compound 7-bromo-5-fluoro-N-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (30 mg, 60 μmol, 20% yield) was obtained as a yellow solid.

[0147] Step 2Pd(OAc)₂ (898 μg, 4.00 μmol, 0.1 equivalent), tri-o-tolylphosphine (1.2 mg, 4.0 μmol, 0.1 equivalent), and TEA (8.1 mg, 80 μmol, 11 μL, 2 equivalent) were added to a solution of 7-bromo-5-fluoro-N-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (30 mg, 39.98 μmol, 1 equivalent) and ethylene sulfonamide (9 mg, 80 μmol, 2 equivalent) in DMF (0.1 mL). The mixture was stirred at 80 °C under N₂ for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge BEH C18 100×30mm×10μm; mobile phase: [H2O(10mM NH4HCO3)-MeCN]; gradient: 30%-60% B, 8.0 min). A yellow solid (E)-2-[5-fluoro-2-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (6 mg, 19% yield, 97% purity) was obtained. 1 H NMR (400MHz, DMSO- d 6) δ = 9.41 (br d, J = 9.6Hz,2H), 7.68 - 7.59 (m, 2H), 7.58 - 7.42 (m, 3H), 7.29 - 7.17 (m, 3H), 6.86 -6.78 (m, 1H), 3.18 (br s, 4H), 2.48 - 2.43 (m, 4H), 2.23 (s, 3H). m / z (ES + [M+1] + = 527.1.

[0148] Prepared according to the general reaction scheme and the synthesis procedure of Example 4, as shown in Table I: Table I

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161] Example 74

[0162] Step 1 A solution of 7-bromo-2-chloro-8-methoxyquinazoline (60 mg, 219 μmol, 1 equivalent), 5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)aniline (60 mg, 219 μmol, 1 equivalent), and TFA (50 mg, 439 μmol, 32.6 μL, 2 equivalent) in n-BuOH (6 mL) was stirred at 90 °C for 12 h. The mixture (combined in 15 mg increments) was concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (neutral conditions; column: Waters XBridge Prep OBD C18 150 × 40 mm × 10 μm; mobile phase: [H2O (10 mM NH4HCO3)-ACN]; gradient: 40%-70% B, for 8.0 min). 20 mg of 7-bromo-8-methoxy-N-(5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)quinazolin-2-amine 5-2 (39 μmol) was obtained as a brown oil.

[0163] Step 2Under N2, Tris-o-tolylphosphine (1.2 mg, 4 μmol, 0.1 equivalent) and Pd(OAc)2 (876 μg, 3.9 μmol, 0.1 equivalent) were added to a solution of 7-bromo-8-methoxy-N-(5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)quinazolin-2-amine (20 mg, 39 μmol, 1 equivalent), ethylene sulfonamide (4 mg, 39 μmol, 1 equivalent), and TEA (8 mg, 78 μmol, 11 μL, 2 equivalent) in DMF (0.5 mL). The mixture was stirred at 100 °C for 12 hours under N2. The reaction mixture (combined in 40 mg increments) was filtered and concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC (neutral conditions; column: Waters XBridge Prep OBD C18 150×40mm×10μm; mobile phase: [H2O (10mM NH4HCO3)-ACN]; gradient: 30%-60% B, for 8.0 min). 17 mg of (E)-2-(8-methoxy-2-((5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)amino)quinazolin-7-yl)ethylene-1-sulfonamide (29 μmol, 93% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ ppm 9.32 (s, 1H), 9.19 (s, 1H), 7.89 (s, 1H),7.74-7.68 (m, 3H), 7.45 (d, J = 8.4Hz, 1H), 7.25-7.22 (m, 3H), 6.78-6.75 (m,1H), 4.02 (s, 3H), 3.21-3.19 (m, 4H), 2.48-2.43 (m, 4H), 2.23 (s, 3H). m / z (ES + [M+1] + = 539.2.

[0164] Example 75

[0165] Step 1A solution of BBr3 (586 mg, 2.34 mmol, 225 μL, 4 equivalents) in DCM (3 mL) was added to a mixture of 7-bromo-2-chloro-8-methoxyquinazoline (0.16 g, 585 μmol, 1 equivalent) in DCM (1.6 mL) at 0 °C under N2. The mixture was stirred at 20 °C for 12 hours. A solution of DCM / MeOH (6 mL, 5:1) was added to the reaction mixture at 0 °C. The mixture was concentrated under reduced pressure to obtain a residue. 7-bromo-2-chloro-quinazoline-8-ol 6-1 (0.2 g, crude) was obtained as a brown solid. m / z (ES + [M+1] + = 259.0.

[0166] Step 2 A mixture of 7-bromo-2-chloroquinazoline-8-ol (0.2 g, 771 μmol, 1 equivalent), K₂CO₃ (213.04 mg, 1.54 mmol, 2 equivalents), and iodoethane (240 mg, 1.54 mmol, 123 μL, 2 equivalents) in DMF (2 mL) was stirred at 60 °C for 2 h. The reaction mixture was quenched by adding H₂O (5 mL) and extracted with EtOAc (5 mL × 3). The combined organic layers were washed with brine (3 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by preparative TLC (SiO₂, petroleum ether / ethyl acetate = 5:1). 7-bromo-2-chloro-8-ethoxy-quinazoline 6-2 (0.15 g, 475 μmol, 62% yield) was given as a yellow solid. m / z (ES + [M+1] + = 287.2.

[0167] Step 3A mixture of 7-bromo-2-chloro-8-ethoxyquinazoline (90 mg, 313 μmol, 1 equivalent), 5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)aniline (86 mg, 313 μmol, 1 equivalent), and TFA (71 mg, 626 μmol, 46.50 μL, 2 equivalent) in n-BuOH (1 mL) was stirred at 90 °C for 12 h. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (neutral conditions; column: Waters XBridge Prep OBD C18 150 × 40 mm × 10 μm; mobile phase: [H2O (10 mM NH4HCO3)-ACN]; gradient: 45%-75% B, for 8.0 min). 7-Bromo-8-ethoxy-N-(5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)quinazolin-2-amine 6-3 was obtained as a brown solid (60 mg, 112 μmol, 36% yield, 98% purity). m / z (ES + [M+1] + = 526.1.

[0168] Step 4 Under N2 conditions, Tris-o-tolylphosphine (3.68 mg, 12.09 μmol, 0.1 equivalent) and Pd(OAc)2 (2.71 mg, 12.1 μmol, 0.1 equivalent) were added to a solution of 7-bromo-8-ethoxy-N-(5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)quinazolin-2-amine (60 mg, 114 μmol, 0.943 equivalent), ethylene sulfonamide (13 mg, 121 μmol, 1 equivalent), and TEA (25 mg, 242 μmol, 34 μL, 2 equivalent) in DMF (1 mL). The mixture was stirred at 100 °C for 12 hours. The reaction was filtered and concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC (neutral conditions; column: WatersXBridge Prep OBD C18 150×40mm×10μm; mobile phase: [H2O (10mM NH4HCO3)-ACN]; gradient: 30%-60% B, for 8.0 min). A yellow solid (E)-2-(8-ethoxy-2-((5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl)amino)quinazolin-7-yl)ethane-1-sulfonamide Example 75 (15 mg, 27.15 μmol, 22.5% yield, 100% purity) was obtained. 1 H NMR (400MHz, CD3CN) δ ppm 9.19 (s, 1H), 8.16 (d, J=2.8Hz, 1H), 7.89 (d, J = 15.6Hz, 1H), 7.73 (s, 1H), 7.62 (d, J = 8.8Hz, 1H), 7.57 (d, J = 7.6Hz, 1H), 7.28-7.23 (m, 2H), 6.72-6.69 (m, 1H), 5.54 (s, 2H), 4.45-4.40 (m, 2H), 3.23 (t, J = 4.8Hz, 4H), 2.52 (t, J = 4.8Hz, 4H), 2.28 (s,3H), 1.33 (t, J = 7.2Hz, 3H). m / z (ES + [M+1] + = 553.2.

[0169] Prepared according to the general reaction scheme and the synthesis procedure of Example 6, as shown in Table I (below): Table I (continued)

[0170] Example 78

[0171] Step 1 A mixture of 5-nitro-2-(trifluoromethoxy)aniline 7-1 (1.3 g, 5.85 mmol, 1 equivalent), 7-bromo-5-fluoro-2-iodo-quinazoline intermediate E (2.07 g, 5.85 mmol, 1 equivalent), Davephos Pd G3 (44.67 mg, 58.53 μmol, 0.01 equivalent), and NaOtBu (2 M, 5.85 mL, 2 equivalent) in dioxane (25 mL) was degassed and purged three times with N2. The mixture was then stirred at 60 °C for 12 hours under N2 atmosphere. The mixture was filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50:1 to 10:1). 0.9 g of 7-bromo-5-fluoro-N-[5-nitro-2-(trifluoromethoxy)phenyl]quinazoline-2-amine as a yellow solid was obtained. m / z (ES + ) [M+H]+= 446.9.

[0172] Step 2Fe (562.02 mg, 10.06 mmol, 5 equivalents) was added to a solution of 7-bromo-5-fluoro-N-[5-nitro-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (0.9 g, 2.01 mmol, 1 equivalent) and NH4Cl (538.34 mg, 10.06 mmol, 5 equivalents) in H2O (2 mL) and EtOH (4 mL) at 50 °C. After addition, the resulting mixture was stirred at 85 °C for 2 hours. The mixture was filtered to obtain a filtrate, which was then concentrated to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 10 / 1). The compound N3-(7-bromo-5-fluoro-quinazolin-2-yl)-4-(trifluoromethoxy)phenyl-1,3-diamine (0.4 g, 958.88 μmol, 48% yield) was given as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ = 9.44(s, 1H), 9.34 (s, 1H), 7.71 (s, 1H), 7.50 (d, J = 9.6Hz, 1H), 7.28 (d, J = 1.6Hz, 1H), 7.09 (d, J = 8.8Hz, 1H), 6.43-6.48 (m, 1H), 5.41 (s, 2H).

[0173] Step 3 To a solution of N3-(7-bromo-5-fluoro-quinazolin-2-yl)-4-(trifluoromethoxy)phenyl-1,3-diamine (100 mg, 239.72 μmol, 1 equivalent), 4A MS (100 mg), and 2-morpholine acetaldehyde (46.44 mg, 359.58 μmol, 1.5 equivalent) in MeOH (2 mL), NaBH3CN (30.13 mg, 479.44 μmol, 2 equivalent) was added. The mixture was stirred at 25 °C for 12 hours. The mixture was filtered to obtain a filtrate, which was then concentrated to remove the solvent. The residue was purified by preparative TLC (SiO2, PE:EtOAc = 1:1). The compound N3-(7-bromo-5-fluoro-quinazolin-2-yl)-N1-(2-morpholinoethyl)-4-(trifluoromethoxy)benzene-1,3-diamine (60 mg, 113.14 μmol, 47% yield) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ =9.39 (s, 1H), 9.35 (s, 1H), 7.63 (s, 1H), 7.44-7.47 (m, 1H), 7.26 (d,J =2.2Hz, 1H), 7.10 (d, J =8.0Hz, 1H), 6.43 (dd, J =2.8, 8.8Hz, 1H), 5.82 (t, J =5.6Hz, 1H), 3.58 (t, J =4.8Hz, 4H), 3.13-3.19 (m, 2H), 2.52-2.54 (m, 2H), 2.40-2.44 (m, 4H).

[0174] Step 4 A mixture of N3-(7-bromo-5-fluoro-quinazolin-2-yl)-N1-(2-morpholinoethyl)-4-(trifluoromethoxy)phenyl-1,3-diamine (50 mg, 94.28 μmol, 1 equivalent), ethylene sulfonamide (15.15 mg, 141.43 μmol, 1.5 equivalent), diacetoxypalladium (423.35 μg, 1.89 μmol, 0.02 equivalent), tri-o-tolylphosphine (1.15 mg, 3.77 μmol, 0.04 equivalent), and TEA (28.62 mg, 282.85 μmol, 39.37 μL, 3 equivalent) in DMF (2 mL) was degassed and purged three times with N2. The mixture was then stirred at 100 °C for 12 hours under N2 atmosphere. The mixture was filtered and concentrated to remove the solvent. The residue was purified by preparative HPLC (neutral conditions; column: CD07-Daisogel SP-100-8-ODS-PK 150×25×10μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 32%-62% B, for 10 min). The compound (E)-2-[5-fluoro-2-[5-(2-morpholinoethylamino)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (4 mg, 7.19 μmol, 8% yield) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ = 9.38 (s, 1H), 9.28 (s, 1H), 7.58-7.69 (m, 2H), 7.49-7.58 (m, 1H), 7.38-7.46 (m, 1H), 7.25 (br s, 2H), 7.18 (d, J = 1.6Hz, 1H), 7.10 (d, J= 9.4Hz, 1H), 6.42-6.46 (m, 1H), 5.78 (t, J =5.2Hz, 1H), 3.57 (t,J = 4.4Hz, 4H), 3.13-3.20 (m, 2H), 2.52-2.55 (m, 2H), 2.40-2.44(m, 4H). m / z (ES + ) [M+H]+= 557.2.

[0175] Prepared according to the teachings of the general reaction scheme and the synthesis procedure of Example 78, as shown in Table I: Table I (continued)

[0176]

[0177] Example 87

[0178] Step 1 The mixture of 5-nitro-2-(trifluoromethoxy)aniline (5 g, 22.51 mmol, 1 equivalent) in HCOOH (50 mL) was degassed and purged three times with N2. The mixture was then stirred at 100 °C for 1 hour under N2 atmosphere. The mixture was concentrated to remove the solvent and used directly in the next step without further purification. The compound N-[5-nitro-2-(trifluoromethoxy)phenyl]formamide (5 g, 19.99 mmol, 89% yield) was given as a dark brown solid. m / z (ES+) [M+H]+ = 251.0.

[0179] Step 2 NaH (11.99 g, 299.84 mmol, 60% purity, 15 equivalents) was added fractionally to a solution of N-[5-nitro-2-(trifluoromethoxy)phenyl]formamide (5 g, 19.99 mmol, 1 equivalent) in THF (60 mL) at 0 °C under N2. The mixture was stirred at 20 °C under N2 for 0.5 h. Then, 7-bromo-2-chloro-5-fluoro-quinazoline (6.27 g, 23.99 mmol, 1.2 equivalents) was added fractionally to the mixture at 0 °C under N2, and the mixture was stirred at 20 °C under N2 for 47.5 h. The reaction mixture was quenched with saturated NH4Cl (20 mL) and treated with EtOAc (10 mL). 3) Extraction. The combined organic layers were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to produce a residue. The crude product was used directly in the next step without purification. A yellow solid, 7-bromo-5-fluoro-N-[5-nitro-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (6 g, crude product), was obtained. m / z (ES+) [M+H]+ = 447.0, 449.0.

[0180] Step 3 Fe (3.75 g, 67.09 mmol, 5 equivalents) was added fractionally to a solution of 7-bromo-5-fluoro-N-[5-nitro-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (6 g, 13.42 mmol, 1 equivalent) and NH4Cl (3.59 g, 67.09 mmol, 5 equivalents) in H2O (20 mL) and EtOH (40 mL) at 50 °C. After addition, the resulting mixture was stirred at 85 °C for 2 hours. The reaction mixture was filtered, and the filtrate was concentrated to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1). The compound N3-(7-bromo-5-fluoro-quinazolin-2-yl)-4-(trifluoromethoxy)phenyl-1,3-diamine (3 g, 7.19 mmol, 54% yield) was given as a yellow solid. 1 H NMR (400MHz, DMSO- d 6) δ = 9.44 (s, 1H), 9.34 (s, 1H), 7.71 (s, 1H), 7.50 (d, J = 9.6Hz, 1H), 7.28 (d, J = 1.6Hz, 1H), 7.09 (d, J = 8.8Hz, 1H), 6.46 (dd, J = 8.4Hz, 2.4Hz, 1H), 5.41 (s, 2H).

[0181] Step 4 The mixture of N3-(7-bromo-5-fluoro-quinazolin-2-yl)-4-(trifluoromethoxy)benzene-1,3-diamine (3 g, 7.19 mmol, 1 equivalent), ethylene sulfonamide (924.53 mg, 8.63 mmol, 1.2 equivalent), Pd(OAc)2 (32.29 mg, 143.83 μmol, 0.02 equivalent), tri-o-tolylphosphine (87.55 mg, 287.66 μmol, 0.04 equivalent), and TEA (2.18 g, 21.57 mmol, 3.00 mL, 3 equivalent) in DMF (30 mL) was degassed and purged three times with N2. The mixture was then stirred at 100 °C for 12 hours under N2 atmosphere. The reaction mixture was filtered and concentrated to remove the solvent. The crude product was milled at 20 °C for 30 minutes with petroleum ether:EtOAc (5:1, 20 mL). The compound (E)-2-[2-[5-amino-2-(trifluoromethoxy)anilino]-5-fluoro-quinazolin-7-yl]ethylenesulfonamide (2.7 g, 6.09 mmol, 85% yield) was obtained as a yellow solid. 1H NMR (400MHz, DMSO-) d 6) δ = 9.38 (s, 1H), 9.23 (s, 1H), 7.65 (s, 1H), 7.61 (d, J = 11.2Hz, 1H), 7.57-7.49 (m, 1H), 7.46-7.37 (m, 1H), 7.25 (br s, 2H), 7.14 (d, J = 2.8Hz, 1H), 7.05-7.01 (m, 1H), 6.43-6.38 (m, 1H), 5.33 (s, 2H).

[0182] Step 5 At 0 °C, a solution of (E)-2-[2-[5-amino-2-(trifluoromethoxy)anilino]-5-fluoro-quinazolin-7-yl]ethylenesulfonamide (500 mg, 1.13 mmol, 1 equivalent) and p-TsOH·H2O (643.53 mg, 3.38 mmol, 3 equivalents) in ACN (12 mL) was added dropwise to a solution of NaNO2 (155.61 mg, 2.26 mmol, 2 equivalents) and KI (468.01 mg, 2.82 mmol, 2.5 equivalents) in H2O (4 mL). The mixture was stirred at 20 °C under N2 for 2 hours. The reaction mixture was quenched by adding H2O (60 mL) and an aqueous solution of Na2S2O3 (2 M, 6 mL), followed by EtOAc (10 mL). 3) Extraction. The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The crude product was ground with EtOAc (10 mL) at 20 °C for 20 minutes. (E)-2-[5-fluoro-2-[5-iodo-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (400 mg, 721.69 μmol, 64% yield) was obtained as a pink solid. 1 H NMR (400MHz, DMSO- d 6) δ = 9.74 (s, 1H), 9.44(s, 1H), 8.36 (d, J = 2.0Hz, 1H), 7.74-7.64 (m, 2H), 7.64-7.59 (m, 1H), 7.55(s, 1H), 7.51-7.44 (m, 1H), 7.28-7.21 (m, 3H).

[0183] Step 6A mixture of 4-bromo-2H-isoquinoline-1-one (400 mg, 1.79 mmol, 1 equivalent), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1,3,2-dioxacyclopentaborane (2.72 g, 10.71 mmol, 6 equivalents), Pd2(dba)3 (163.48 mg, 178.53 μmol, 0.1 equivalents), tricyclohexylphosphine (100.13 mg, 357.06 μmol, 0.2 equivalents), and AcOK (525.64 mg, 5.36 mmol, 3 equivalents) in dioxane (20 mL) was degassed and purged three times with N2. The mixture was then stirred at 110 °C for 12 hours under N2 atmosphere. The reaction mixture was filtered to obtain a filtrate, which was then concentrated to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1). The compound 4-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-2H-isoquinoline-1-one (300 mg, 1.11 mmol, 62% yield) was given as a white solid. m / z (ES+) [M+H]+ = 272.1.

[0184] Step 7 The following ingredients were added: (E)-2-[5-fluoro-2-[5-iodo-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylene sulfonamide (50 mg, 90.21 μmol, 1 equivalent), 4-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-2H-isoquinolin-1-one (36.69 mg, 135.32 μmol, 1.5 equivalent), and Pd(dppf)Cl2. . CH₂Cl₂ (7.37 mg, 9.02 μmol, 0.1 equivalent) and Na₂CO₃ (23.90 mg, 225.53 μmol, 2.5 equivalent) were degassed in a mixture of THF (2 mL) and H₂O (0.2 mL) and purged three times with N₂. The mixture was then stirred at 70 °C for 12 hours under N₂ atmosphere. The reaction mixture was filtered and washed with DMF (3 mL), and the filtrate was concentrated to remove the solvent. The residue was subjected to preparative HPLC (column: CD07-Daisogel SP-100-8-ODS-PK 150). 25 10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 35%-65% B (for 15 minutes) purification. The compound (E)-2-[5-fluoro-2-[5-(1-oxo-2H-isoquinolin-4-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (3 mg, 5.25 μmol, 6% yield) was obtained as a yellow solid.1 H NMR (400MHz, DMSO- d 6)δ = 11.53 (br s, 1H), 9.80 (s, 1H), 9.43 (s, 1H), 8.32 (d, J = 8.0Hz, 1H),8.05 (s, 1H), 7.82-7.73 (m, 2H), 7.65-7.61 (m, 2H), 7.60-7.52 (m, 3H), 7.47-7.40 (m, 1H), 7.38-7.33 (m, 1H), 7.30-7.19 (m, 3H). m / z (ES+) [M+H]+= 572.1.

[0185] Prepared according to the teachings of the general reaction scheme and the synthesis procedure of Example 87, as shown in Table I: Table I (continued)

[0186] Example 90

[0187] Step 1: NBS (495.49 mg, 2.78 mmol, 1 equivalent) was added fractionally to a solution of 6-chloro-2H-isoquinoline-1-one (500 mg, 2.78 mmol, 1 equivalent) in THF (9 mL) and DCM (6 mL) at 0 °C. The mixture was stirred at 20 °C under N2 for 12 hours. The reaction mixture was quenched by adding H2O (20 mL), extracted with CH2Cl2 (10 mL × 3), and washed with 30 mL of brine (10 mL × 3). The organic phases were then combined, dried over Na2SO4, filtered, and concentrated under vacuum to give the residue. The crude product was milled at 20 °C with DCM (10 mL) for 20 minutes. The compound 4-bromo-6-chloro-2H-isoquinoline-1-one (450 mg, 1.74 mmol, 63% yield) was given as a white solid. 1H NMR (400MHz, DMSO-d6) δ = 11.73 (s, 1H), 8.22 (d, J = 8.8Hz, 1H), 7.71 (d, J = 2.0Hz, 1H), 7.67-7.60 (m, 2H).

[0188] Step 2: Degas the mixture of 4-bromo-6-chloro-2H-isoquinoline-1-one (400 mg, 1.55 mmol, 1 equivalent), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1,3,2-dioxacyclopentaborane (707.30 mg, 2.79 mmol, 1.8 equivalent), Pd2(dba)3 (141.70 mg, 154.74 μmol, 0.1 equivalent), tricyclohexylphosphine (86.79 mg, 309.48 μmol, 0.2 equivalent), and KOAc (455.59 mg, 4.64 mmol, 3 equivalent) in dioxane (10 mL) and purge with N2 three times. Then, stir the mixture at 110 °C for 12 hours under N2 atmosphere. The reaction mixture was filtered to obtain a filtrate, which was then concentrated to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1). The compound 6-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-2H-isoquinoline-1-one (360 mg, 1.18 mmol, 76% yield) was given as a white solid. m / z (ES+) [M+H]+ = 306.1, 308.0.

[0189] Step 3: Add 6-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-2H-isoquinoline-1-one (106.02 mg, 270.63 μmol, 1.5 equivalent), Pd(dppf)Cl2.CH2Cl2 (14.73 mg, 18.04 μmol, 0.1 equivalent) and Na2CO3 (47.81 mg, 451.05 μmol, 2.5 equivalent) to a solution of (E)-2-[5-fluoro-2-[5-iodo-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (100 mg, 180.42 μmol, 1 equivalent) in THF (2 mL) and water (0.2 mL). The mixture was degassed and purged three times with N2, and then stirred at 70°C for 12 hours under N2 atmosphere.

[0190] The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was then passed through a preparative HPLC system (column: CD07-Daisogel SP-100-8-ODS-PK 150). 25 10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 40%-70% B (after 15 min) purification. The compound (E)-2-[2-[5-(6-chloro-1-oxo-2H-isoquinolin-4-yl)-2-(trifluoromethoxy)anilino]-5-fluoro-quinazolin-7-yl]ethylenesulfonamide (4 mg, 6.46 μmol, 98% purity) was obtained as a yellow solid. 1H NMR(400MHz, DMSO-d6) δ = 11.83-11.49 (m, 1H), 9.82(s, 1H), 9.44(s, 1H), 8.32 (d,J = 8.4Hz, 1H), 8.06 (d, J = 2.0Hz, 1H), 7.69-7.60 (m, 4H), 7.58-7.50 (m,2H), 7.44-7.38 (m, 1H), 7.35 (dd, J = 8.4Hz, 2.0Hz, 1H), 7.33-7.16 (m, 3H). m / z (ES+) [M+H]+ = 606.0, 608.0.

[0191] Example 91

[0192] Step 1 TsOH (216.35 mg, 1.26 mmol, 0.2 equivalent) was added to a solution of 6-methyl-2H-isoquinoline-1-one (1 g, 6.28 mmol, 1 equivalent) and NIS (2.12 g, 9.42 mmol, 1.5 equivalent) in DCE (6 mL) under N2. The mixture was stirred at 25 °C for 12 h. The reaction mixture was quenched by adding H2O (20 mL) and then extracted with DCM (20 mL × 2). The combined organic layers were dried over Na2SO4 and filtered, and the filtrate was concentrated under reduced pressure to produce a residue. The crude product was milled with DCM (10 mL) for 15 min. The compound 4-iodo-6-methyl-2H-isoquinoline-1-one (400 mg, 1.40 mmol, 22% yield) was given as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 11.41 (s,1H), 8.08 (d, J =8.0Hz, 1H), 7.58 (d, J = 4.4Hz, 1H), 7.40(t, J = 8.0Hz, 2H), 2.54 (s, 3H). m / z(ES+) [M+H]+ = 285.9 Step 2 A mixture of 5-nitro-2-(trifluoromethoxy)aniline (5 g, 22.51 mmol, 1 equivalent) in HCOOH (100 mL) was degassed and purged three times with N2. The mixture was then stirred at 100 °C for 1 hour under N2 atmosphere. The compound N-[5-nitro-2-(trifluoromethoxy)phenyl]formamide (16 g, 63.97 mmol, 95% yield) was given as a brown solid. m / z(ES+) [M+H]+ = 251.0.

[0193] Step 3 NaH (11.99 g, 299.84 mmol, 60% purity, 15 equivalents) was added fractionally to a solution of N-[5-nitro-2-(trifluoromethoxy)phenyl]formamide (5 g, 19.99 mmol, 1 equivalent) in THF (60 mL) at 0 °C under N2. The mixture was stirred at 20 °C under N2 for 0.5 h. Then, 7-bromo-2-chloro-5-fluoro-quinazoline (5.75 g, 21.99 mmol, 1.1 equivalents) was added fractionally to the mixture at 0 °C under N2, and the mixture was stirred at 20 °C under N2 for 47.5 h. The reaction mixture was quenched with saturated NH4Cl (60 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were dried over Na2SO4 and filtered, and the filtrate was concentrated under reduced pressure to produce a residue. The obtained compound was used directly in the next step without further purification. The compound 7-bromo-5-fluoro-N-[5-nitro-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (4 g, crude) was obtained as a yellow solid. This compound was used directly in the next step without any purification.

[0194] Step 4 Fe (1.25 g, 22.36 mmol, 5 equivalents) was added fractionally to a solution of 7-bromo-5-fluoro-N-[5-nitro-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (2 g, 4.47 mmol, 1 equivalent) and NH4Cl (1.20 g, 22.36 mmol, 5 equivalents) in H2O (20 mL) and EtOH (40 mL) at 85 °C. After addition, the resulting mixture was stirred at 85 °C for 2 hours. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1). The compound N3-(7-bromo-5-fluoro-quinazolin-2-yl)-4-(trifluoromethoxy)phenyl-1,3-diamine (3 g, 7.19 mmol, 54% yield) was given as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ = 9.44 (s,1H), 9.34 (s, 1H), 7.71 (s, 1H), 7.50 (d, J = 9.6Hz, 1H), 7.28 (d, J = 1.6Hz,1H), 7.09 (d, J = 8.8Hz, 1H), 6.46 (dd, J = 2.4Hz, 8.4Hz, 1H), 5.41 (s, 2H).

[0195] Step 5 A mixture of N3-(7-bromo-5-fluoro-quinazolin-2-yl)-4-(trifluoromethoxy)phenyl-1,3-diamine (3.0 g, 7.20 mmol, 1 equivalent), ethylene sulfonamide (924.54 mg, 8.62 mmol, 1.2 equivalent), Pd(OAc)2 (32.3 mg, 143.84 μmol, 0.02 equivalent), tri-o-tolylphosphine (87.56 mg, 287.66 μmol, 0.04 equivalent), and TEA (2.18 g, 21.58 mmol, 3.00 mL, 3 equivalent) in DMF (60 mL) was degassed and purged three times with N2. The mixture was then stirred at 100 °C for 12 hours under N2 atmosphere. The mixture was filtered to obtain a filtrate, which was then concentrated under reduced pressure to obtain a residue. The crude product was ground with PE:EtOAc (5:1, 20 mL) at 20 °C for 30 minutes and filtered to obtain (E)-2-[2-[5-amino-2-(trifluoromethoxy)anilino]-5-fluoro-quinazolin-7-yl]ethylenesulfonamide (2.7 g, 6.09 mmol, 85% yield) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 9.38 (s, 1H), 9.23 (s, 1H), 7.65 (s, 1H), 7.61(d, J = 11.2Hz, 1H), 7.57-7.49 (m, 1H), 7.46-7.37 (m, 1H), 7.25 (br s, 2H), 7.14 (d, J = 2.8Hz, 1H), 7.03 (dd, J = 1.2Hz, 8.8Hz, 1H), 6.41 (dd, J =2.8Hz, 8.8Hz, 1H), 5.33 (s, 2H).

[0196] Step 6At 0 °C, a solution of (E)-2-[2-[5-amino-2-(trifluoromethoxy)anilino]-5-fluoro-quinazolin-7-yl]ethylenesulfonamide (1 g, 2.26 mmol, 1 equivalent) and p-TsOH·H₂O (1.29 g, 6.77 mmol, 3 equivalents) in ACN (18 mL) was added dropwise with NaNO₂ (311.23 mg, 4.51 mmol, 2 equivalents) and KI (936.02 mg, 5.64 mmol, 2.5 equivalents) in H₂O (6 mL). The mixture was stirred at 20 °C for 2 hours. The reaction mixture was quenched by adding H₂O (120 mL) and Na₂S₂O₃ (2 M, 10 mL), followed by extraction with EtOAc (20 mL × 3). The combined organic layers were dried over Na₂SO₄ and filtered, and the filtrate was concentrated under reduced pressure to give the residue. The crude product was ground with EtOAc:PE (5:1, 20 mL) at 20 °C for 30 minutes and filtered to obtain (E)-2-[5-fluoro-2-[5-iodo-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (700 mg, 1.26 mmol, 28% yield) as a dark red solid. 1 H NMR (400MHz, DMSO-d6) δ = 9.74 (s,1H), 9.44 (s, 1H), 8.36 (dd, J = 2.0Hz, 1H), 7.74-7.64 (m, 2H), 7.64-7.59 (m,1H), 7.56 (s, 1H), 7.53 (m, 1H), 7.28-7.21 (m, 3H).

[0197] Step 7 A mixture of (E)-2-[5-fluoro-2-[5-iodo-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (500 mg, 902.11 μmol, 1 equivalent), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)-1,3,2-dioxacyclopentaborane (458.16 mg, 1.80 mmol, 2 equivalents), cataCXiumA Pd G3 (65.70 mg, 90.21 μmol, 0.1 equivalents), and AcOK (265.61 mg, 2.71 mmol, 3 equivalents) in dioxane (10 mL) was degassed and purged three times with N2. The mixture was then stirred at 90 °C for 12 hours under N2 atmosphere. The mixture was filtered to obtain a filtrate, which was then concentrated under reduced pressure to obtain a residue. The residue was passed through preparative HPLC (neutral conditions; column: CD02-Waters Xbidge BEH C18 150). 25 10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 52%-72% B (after 15 min) purification. The compound [3-[[5-fluoro-7-[(E)-2-aminosulfonylvinyl]quinazolin-2-yl]amino]-4-(trifluoromethoxy)phenyl]boronic acid (150 mg, 317.68 μmol, 35% yield) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ= 9.64 (s, 1H), 9.39 (s, 1H), 8.21 (s, 2H), 8.09 (d, J = 1.2Hz, 1H), 7.72(dd, J- = 1.6Hz, 8.4Hz, 1H), 7.62-7.57 (m, 2H), 7.52 (s, 1H), 7.44 (s, 1H), 7.40-7.36 (m, 1H), 7.24 (s, 2H).

[0198] Step 8 Pd(dppf)Cl2 was added to a solution of [3-[[5-fluoro-7-[(E)-2-aminosulfonylvinyl]quinazolin-2-yl]amino]-4-(trifluoromethoxy)phenyl]boronic acid (62.11 mg, 131.54 μmol, 1.5 equivalents) and 4-iodo-6-methyl-2H-isoquinolin-1-one (25 mg, 87.69 μmol, 1 equivalent) in THF (1 mL) and H2O (0.1 mL) under N2 conditions. . CH₂Cl₂ (7.16 mg, 8.77 μmol, 0.1 equivalent) and Na₂CO₃ (27.88 mg, 263.08 μmol, 3 equivalent). The mixture was stirred at 75 °C for 6 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to provide the residue. The residue was subjected to preparative HPLC (column: CD03-Welch Ultimate C18 150). 25 Purification was performed using a mobile phase of 5 μm (water (HCl)-ACN) and a gradient of 40%-70% B for 10 minutes. The compound (E)-2-[5-fluoro-2-[5-(6-methyl-1-oxo-2H-isoquinolin-4-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (8 mg, 12.86 μmol, 15% yield) was obtained as a yellow solid. 1H NMR(400MHz, DMSO-d6) δ = 11.45 (d, J=6.0Hz,1H), 9.79 (s, 1H), 9.43 (s, 1H), 8.22(d, J=8.0Hz,1H), 8.027 (d, J=2.0Hz,1H), 7.65-7.59 (m, 2H), 7.55-7.51 (m, 2H),7.47 (s, 1H), 7.43-7.41 (m, 1H), 7.39 (s, 1H), 7.36 (dd, J=2.0Hz,8.4Hz,1H),7.26 (s,2H), 7.19 (d, J=6.0Hz, 1H), 2.42 (s, 3H). m / z (ES+) [M+H]+ = 586.1 Prepared according to the teachings of the general reaction scheme and the synthesis procedure of Example 91, as shown in Table I: Table I (continued)

[0199] Example 98

[0200] Step 1 The mixture of (E)-2-[5-fluoro-2-[5-iodo-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (intermediate H, 80 mg, 144.34 μmol, 0.7 equivalents), 1-cyclopropylpiperazine (39.03 mg, 309.29 μmol, 1.3 equivalents), Pd-PEPPSI-IHept-Cl (20.06 mg, 20.62 μmol, 0.1 equivalents), and t-BuONa (2 M, 309.29 μL, 3 equivalents) in 2-methylbut-2-ol (2 mL) was degassed and purged three times with N2. The mixture was then stirred at 90 °C for 12 hours under N2 atmosphere. The mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was subjected to preparative HPLC (column: CD07-Daisogel SP-100-8-ODS-PK 150). 25 10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 44%-74% B (for 11 min) purification. The compound (E)-2-[2-[5-(4-cyclopropylpiperazin-1-yl)-2-(trifluoromethoxy)anilino]-5-fluoro-quinazolin-7-yl]ethylenesulfonamide (12 mg, 20.68 μmol, 96% yield, 95.24% purity) was obtained as a yellow solid. 1H NMR (400MHz, DMSO- d 6) δ = 9.42 (s, 1H), 9.40 (s, 1H), 7.69-7.60 (m, 2H), 7.59-7.42 (m, 3H), 7.29-7.19 (m, 3H), 6.81 (dd, J = 2.8Hz, 9.2Hz, 1H), 3.14(t, J = 4.4Hz, 4H), 2.69 (t, J = 1.2Hz, 4H), 1.73-1.61 (m, 1H), 0.50-0.39 (m, 2H), 0.39-0.29 (m, 2H). m / z (ES+) [M+H]+= 553.3.

[0201] Example 99

[0202] Step 1 The mixture of (E)-2-[5-fluoro-2-[5-iodo-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (80.10 mg, 144.51 μmol, 1.5 equivalents), 1,3-dihydroimidazol-2-one (8.1 mg, 96.34 μmol, 1 equivalent), Pd(OAc)2 (1.08 mg, 4.82 μmol, 0.05 equivalents), and NaOAc (23.71 mg, 289.02 μmol, 3 equivalents) in DMSO (3 mL) was degassed and purged three times with N2. The mixture was then stirred at 80 °C for 12 hours under N2 atmosphere. The reaction mixture was filtered to obtain a filtrate and concentrated to remove the solvent. The residue was subjected to preparative HPLC (column: CD02-Waters Xbidge BEHC18 150). 25 10 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 22%-52% B (for 10 min) purification. The compound (E)-2-[5-fluoro-2-[5-(2-oxo-1,3-dihydroimidazol-4-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (5 mg, 9.40 μmol, 6.0% yield, 96% purity) was obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ = 10.55 (s, 1H), 10.14 (s, 1H), 9.64 (s, 1H), 9.41 (s, 1H), 7.99 (s, 1H), 7.69 (s, 1H), 7.65 (d, J =10.8Hz, 1H), 7.58 (m, 1H), 7.43 (m, 3H), 7.24 (s, 2H), 6.96-6.95 (m, 1H). 1 H NMR (400MHz, DMSO-d6 and a drop of D2O) δ =9.39 (s, 1H), 7.99 (s, 1H), 7.63 (br d, J = 6.3Hz, 1H), 7.55 - 7.49 (m, 1H),7.48 - 7.44 (m, 1H), 7.40-7.34 (m, 1H), 7.38 (br s, 2H), 6.96 - 6.88 (m, 1H). m / z (ES+) [M+H]+ = 511.0 Example 100

[0203] Step 1: Add 4-bromo-2,3,6-trifluorobenzaldehyde (1.8 g, 7.53 mmol, 1 equivalent) and guanidine carbonate (1.09 g, 6.03 mmol, 0.8 equivalent) to a solution of DMA (20 mL). Stir the mixture at 140 °C for 4 hours. Add water (50 mL) to the mixture and filter to obtain a filter cake. Dry the filter cake under vacuum to obtain the desired product. The product is used in the next step without purification. A white solid, 7-bromo-5,8-difluoro-quinazoline-2-amine (1.3 g, 5.00 mmol, 66% yield), is obtained. 1 H NMR (400MHz, DMSO-d6) δ = 9.28 (d, J = 1.0Hz, 1H), 7.62 (br s,2H), 7.36 (dd, J = 4.8, 8.8Hz, 1H).

[0204] Step 2: Tetrabutylammonium chloride (1.03 g, 3.69 mmol, 1.03 mL, 1.2 equivalents) was added to a solution of 7-bromo-5,8-difluoro-quinazolin-2-amine (800 mg, 3.08 mmol, 1 equivalent) in DCM (8 mL) and DMF (1 mL) at 0 °C and maintained for 0.5 h. Then, chloro(trimethyl)silane (1.34 g, 12.31 mmol, 1.56 mL, 4 equivalents) and tert-butyl nitrite (951.74 mg, 9.23 mmol, 1.10 mL, 3 equivalents) were added to the mixture. The mixture was stirred at 45 °C for 4.5 h. The reaction mixture was poured into ice water (50 mL) and extracted with EtOAc (20 mL × 3). The combined organic phases were washed with saturated NaHCO3 (20 mL) and brine (20 mL), dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 10 / 1, 3 / 1). The compound 7-bromo-2-chloro-5,8-difluoro-quinazoline (320 mg, 1.15 mmol, 37% yield) was given as a yellow oil. 1 H NMR (400MHz, DMSO-d6) δ = 9.78 (s, 1H), 8.12 (br dd, J =4.4, 8.8Hz, 1H).

[0205] Step 3: At 0°C under N2, add participles of NaH (161.00 mg, 5.37 mmol, 1.34 mL, 80% purity, 15 equivalents) to a solution of N-[5-(4-methylpiperazin-1-yl-2-(trifluoromethoxy)phenyl]formamide (130.22 mg, 429.39 μmol, 1.2 equivalents) and 7-bromo-2-chloro-5,8-difluoro-quinazoline (100 mg, 357.82 μmol, 1 equivalent) in THF (1 mL). Stir the mixture at 25°C for 48 hours. Add saturated NH4Cl (20 mL) to the mixture. Then, use ethyl acetate (10 mL) to... 3) Extraction of the mixture. The combined organic phases were dried over anhydrous Na₂SO₄, filtered, and concentrated to obtain the residue. The product was used in the next step without purification. The compound 7-bromo-5,8-difluoro-N-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (20 mg, 38.59 μmol, 11% yield) was obtained as a white solid. 1H NMR (400MHz, DMSO-d6) δ =9.80 (s, 1H), 9.45 (d, J = 1.0Hz, 1H), 7.72 (br s, 1H), 7.57 (dd, J = 4.6,8.9Hz, 1H), 7.24 (dd, J = 1.1, 9.0Hz, 1H), 6.83 (dd, J = 2.9, 9.1Hz, 1H), 3.22 - 3.17 (m, 4H), 2.48 (br s, 4H), 2.25 (s, 3H).

[0206] Step 4: To a solution of 7-bromo-5,8-difluoro-N-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]quinazolin-2-amine (70 mg, 135.06 μmol, 1 equivalent) and ethylene sulfonamide (17.36 mg, 162.08 μmol, 1.2 equivalent) in DMF (1 mL), add TEA (41.00 mg, 405.19 μmol, 56.40 μL, 3 equivalent), Pd(OAc)2 (606.45 μg, 2.70 μmol, 0.02 equivalent), and tri-o-tolylphosphine (1.64 mg, 5.40 μmol, 0.04 equivalent). Stir the mixture at 100 °C under N2 for 12 hours. Filter the mixture and concentrate the filtrate to obtain the residue. The residue was passed through a preparative HPLC system (column: Waters Xbridge BEH C18 100). 30mm 10 μm; mobile phase: [H2O(10 mM NH4HCO3)-ACN]; gradient: 45%-75% B, for 8.0 min) purification. The compound (E)-2-[5,8-difluoro-2-[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (1.2 mg, 2.19 μmol, 2% yield, 99% purity) was obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ = 9.73 (s, 1H), 9.45 (s, 1H), 7.77 (s, 1H), 7.73 - 7.68 (m, 1H), 7.67 - 7.61 (m, 1H), 7.59 - 7.52 (m, 1H), 7.38 (s, 2H), 7.24 (br d, J = 8.9Hz, 1H), 6.82 (dd, J = 2.6, 9.1Hz, 1H), 3.20 (br s, 4H), 2.48 (br s, 4H), 2.24 (s, 3H). m / z (ES+) [M+H]+ =545.1.

[0207]

[0208] Example 101 Step 1: TsOH·H₂O (65.58 mg, 344.77 μmol, 0.07 equivalent) and NIS (1.86 g, 8.27 μmol, 1.68 equivalent) were added to a solution of 5-bromo-2-iodo-2-methoxy-pyridine-4-amine (1 g, 4.93 mmol, 1 equivalent) in ACN (20 mL). The mixture was stirred at 70 °C for 12 h. The reaction mixture was concentrated under reduced pressure to obtain a residue. The crude product was milled with EtOAc (20 mL) at 25 °C for 20 min, filtered, and the filter cake was dried under vacuum. The compound 5-bromo-3-iodo-2-methoxy-pyridine-4-amine (1.37 g, 4.16 mmol, 84% yield) was given as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 7.89 (s, 1H), 6.09 (s, 2H), 3.80 (s, 3H). m / z (ES+) [M+H]+ = 328.8, 330.9.

[0209] Step 2: Under N2, Pd(OAc)2 (46.75 mg, 208.25 μmol, 0.05 equivalent) was added to a solution of 5-bromo-3-iodo-2-methoxy-pyridine-4-amine (1.37 g, 4.16 mmol, 1 equivalent), ethyl propionate (750.56 mg, 7.50 mmol, 814.94 μL, 1.8 equivalent), TEA (632.18 mg, 6.25 mmol, 869.57 μL, 1.5 equivalent), and tri-o-tolylphosphine (126.77 mg, 416.50 μmol, 0.1 equivalent) in DMF (14 mL). The mixture was stirred under N2 at 100 °C for 12 hours. The reaction mixture was quenched by adding 30 mL of aqueous NaClO solution, and the mixture was concentrated under reduced pressure to produce a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1). Ethyl (E)-3-(4-amino-5-bromo-2-methoxy-3-pyridyl)prop-2-enoate was given as a yellow solid (0.9 g, 2.99 mmol, 72% yield). 1 H NMR (400MHz, DMSO-d6) δ = 7.95 (s, 1H), 7.72 (d, J =4.0Hz, 1H), 6.63-6.56 (m, 3H), 4.18 (dd, J =6.8Hz, 14.4Hz, 2H), 3.86 (s, 3H), 1.25 (t, J =6.8Hz, 3H). m / z (ES+) [M+H]+ =301.0, 302.8.

[0210] Step 3: Under N2, add NaSMe (230.42 mg, 3.29 mmol, 209.47 μL, 1.1 equivalent) to a solution of ethyl (E)-3-(4-amino-5-bromo-2-methoxy-3-pyridyl)prop-2-enoate (900 mg, 2.99 mmol, 1 equivalent) in EtOH (10 mL). Stir the mixture at 25 °C for 2 hours. Quench the reaction mixture by adding H2O (30 mL) at 0 °C, followed by adding DCM (30 mL). 2) Extraction. The combined organic layers were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 8 / 1). The compound 8-bromo-5-methoxy-1H-1,6-naphthid-2-one (230 mg, 901.72 μmol, 30% yield) was given as a yellow solid. 1HNMR (400MHz, DMSO-d6) δ = 11.14 (s, 1H), 8.30 (s, 1H), 7.99 (d, J =8.8Hz, 1H), 6.60 (d, J =8.8Hz, 1H), 3.98 (s, 3H). m / z (ES+) [M+H]+ = 255.0, 257.0.

[0211] Step 4: Under N2, add Pd(dppf)Cl2.CH2Cl2 (7.37 mg, 9.02 μmol, 0.1 equivalent) to a solution of (E)-2-[5-fluoro-2-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (50 mg, 90.20 μmol, 1 equivalent), 8-bromo-5-methoxy-1H-1,6-naphthidium-2-one (20.71 mg, 81.18 μmol, 0.9 equivalent) and Na2CO3 (28.68 mg, 270.60 μmol, 3 equivalent) in THF (2 mL) and H2O (0.4 mL). Stir the mixture under N2 at 75 °C for 6 hours. Concentrate the reaction mixture under reduced pressure to obtain the residue. The residue was passed through a preparative HPLC system (column: CD24-XPT C18 150). 25 7 μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 35%-65% B (for 10 min) purification. The compound (E)-2-[5-fluoro-2-[5-(5-methoxy-2-oxo-1H-1,6-naphthid-8-yl)-2-(trifluoromethoxy)anilino]quinazolin-7-yl]ethylenesulfonamide (16 mg, 26.56 μmol, 30% yield) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 10.63 (s, 1H), 9.83 (s, 1H), 9.49(s, 1H), 8.19 (d, J =1.6Hz, 1H), 8.11-8.10 (m, 2H), 7.79 (s, 1H), 7.64-7.58 (m,2H), 7.56-7.54 (m, 1H), 7.46-7.42 (m, 1H), 7.33 (dd, J =1.2Hz, 8.8Hz, 1H), 7.27(s, 2H), 6.63 (d, J=9.6Hz, 1H), 4.04 (s, 3H). m / z (ES+) [M+H]+ = 603.1.

[0212] Example A This embodiment illustrates that the exemplary compounds of the present invention inhibit PLK1 enzyme activity.

[0213] The compounds were dissolved in DMSO to prepare a 10 mM stock solution, which was then diluted in DMSO to a 100 μM working solution. A series of ten ternary dilutions of each compound's working solution were dispensed using two technical replicates (Tecan D300e digital dispenser) on a 384-well white ProxiPlate (PerkinElmer, #6008280) to produce a series of dilutions with final concentrations ranging from 0.05 nM to 1000 nM in a total reaction volume of 10 μL. Volasertib (MedChemExpress, #HY-12137) and GSK461364 (MedChemExpress, #HY-50877) were used as positive controls (1.0 μM final concentration, 5 technical replicates each). Kinase buffer (PerkinElmer, HTRF) supplemented with 1.0 mM DTT and 5.0 mM MgCl2 was used. ® For the KinEASE-STKS1 kit (#62ST1PEB), bring the volume to 4 μL and use the same assay buffer without the compound as a negative control. Normalize DMSO to a final concentration of 1% on all wells.

[0214] All subsequently measured fractions were diluted with kinase buffer supplemented with 1 mM DTT and 5 mM MgCl2. A 25 nM working solution of LK1 protein (Carna Biosciences, #05-157) was prepared and 2 μL was dispensed into each well to obtain a final concentration of 5 nM. A 10 μM STK substrate 1-biotin solution (PerkinElmer, HTRF) was prepared according to the manufacturer's instructions. ® KinEASE-STKS1 kit (#62ST1PEB) was used, and 2 μL was dispensed into each well to obtain a final concentration of 2 μM. Finally, a 100 μM ATP solution (Sigma-Aldrich, #A7699) was prepared and added to each well to obtain a final concentration of 20 μM, initiating the kinase reaction. The plate was sealed, briefly shaken to thoroughly mix the reaction contents, centrifuged, and incubated at 37°C for 90 minutes.

[0215] After the kinase reaction is complete, remove the plate seal and proceed with the assay according to the manufacturer's instructions (PerkinElmer, HTRF). ®Prepare 0.5 μM streptavidin-XL665 (PerkinElmer, HTRF) in KinEASE-STK S1 kit #62ST1PEB. ® KinEASE-STK S1 Kit #62ST1PEB working solution was dispensed into each well, yielding a final concentration of 0.125 μM. Then, 5 μL of the antibody-crystallization compound (PerkinElmer, HTRF) was added to each well. ® KinEASE-STK S1 kit #62ST1PEB solution. Reseal the plate, briefly vortex, centrifuge, and incubate at room temperature (25°C) for 60 minutes. Then, incubate with Pherastar. ® FRET signals were read using 620nm / 665nm wavelengths for excitation / emission on an FSX microplate reader (BMG LABTECH). Signal values ​​were normalized to background signal, and the acceptor / donor signal ratio was calculated. GraphPad Prism™ software was used to plot the compound concentration-response curve (CRC) against compound concentration to generate a semi-logarithmic concentration-response curve. The IC50 was determined using nonlinear regression with a variable slope model. 50 value.

[0216] Table II

[0217] Table II (continued)

[0218] Example B This example illustrates the reactivity of glutathione (GSH) in its reduced state at pH 7.5 with the exemplary compounds of the present invention.

[0219] The reactivity of the compound with GSH (USP, catalog 1294820) was analyzed in a time-dependent experiment using liquid chromatography-mass spectrometry (LC-MS). The peak area of ​​the compound was calculated using the peak area normalized with the internal standard (IS) Rhodamine B (Sigma Aldrich, catalog R6626) to determine the half-life (t½).

[0220] The compounds were dissolved in DMSO to obtain a 10 mM concentration. A dilution of each compound was prepared in acetonitrile (LC-MS grade) to obtain a 100 μM concentration. The assay buffer (50 mM Tris, pH 7.5, with 150 mM NaCl) was deoxygenated by maintaining it under a nitrogen flow for approximately 1 hour prior to testing. GSH and IS were dissolved in the assay buffer to obtain 1.2 mM and 25 μM concentrations, respectively.

[0221] 4 μL of 25 μM Rhodamine B solution was added to 86 μL of 1.2 mM GSH solution in a glass vial containing an insert. To initiate the reaction, 10 μL of 100 μM compound solution was added to the vial, and the reaction mixture was immediately injected into the LC-MS system, followed by repeated injections at fixed time intervals for up to 24 hours. The autosampler temperature was set to 25 °C to maintain a stable incubation temperature. The reaction was analyzed using an ACQUITY UPLC system on a Dionex UltiMate 3000 system (ThermoScientific). ® Compounds were separated on an HSS T3 1.8µm, 2.1×100mm column (Waters) at 40 °C using a 10%–95% gradient of 0.1% formic acid in acetonitrile at a flow rate of 0.25 mL / min for 5.5 min. The separation was performed using a Q Exactive ionizer equipped with an electrospray ionization source. ™ The analysis was performed using a Plus mass spectrometer (Thermo Scientific). This mass spectrometer operates in standard full MS mode within a scan range of 200 to 600 m / z.

[0222] Table III

[0223] Table III (continued)

[0224] Based on the obtained chromatogram, Chromeleon was used. ™ The chromatographic data system integrates the peak areas of the test compound and the internal standard (IS), calculates the area ratio between the test compound and the IS, and then calculates the natural logarithm (ln) from the obtained area ratio. The t½ value for each compound is determined by plotting ln (area ratio) against time and the equation t½ = lN² / (-slope).

Claims

1. A compound of formula (I) and a pharmaceutically acceptable salt thereof: Formula (I) in: R 1 It can be hydrogen, halogen, hydroxyl, C1-C6 alkyl, cycloalkyl, haloalkyl, or aniline; R 2 It is a hydrogen, alkoxy, or -O-haloalkyl group optionally substituted with one or more alkoxy groups. R 3 Hydrogen, optionally with one or more R 4 Substituted heterocyclic groups, -NH-L 1 -N(R A R B -NH-L 2 -cycloalkyl or -NH-L 2 - Heterocyclic group, wherein the cycloalkyl group or the heterocyclic group is optionally surrounded by one or more R groups. 5 replace; R 6 It can be OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azacyclobutane, or S(O)(O)CH3; R 7 It can be NH2, NHCH3, or CH3; Each R 4 Independently C1-C4 alkyl, cyclopropyl, hydroxyalkyl, -L 1 -N(R A R B ), halogen, O, O-CH3, NH2, -C(O)O-tert-butyl; -L 3 -aryl; or -L 4 - Heterocyclic group; Each R 5 Independently, it is a C1-C4 alkyl group, N(CH3)(CH3) or a heterocyclic group optionally substituted with an aryl group; Each L 1 It is a C1-C4 alkylene group; Each L 2 It is a bond or a C1-C4 alkylene group; Each L 3 It is -CH2-O-CH2-; Each L 4 For bond or methylene; Each R A It is hydrogen or C1-C3 alkyl; Each R B It is hydrogen or C1-C3 alkyl.

2. The compound according to claim 1, wherein R 1 It is a halogen.

3. The compound according to claim 2, wherein the halogen is chlorine, fluorine or bromine.

4. The compound according to claim 1, wherein R 1 It is a hydroxyl group.

5. The compound according to claim 1, wherein R 1 It is a C1-C6 alkyl group.

6. The compound according to claim 5, wherein the C1-C6 are alkyl groups, specifically methyl, ethyl, or isopropyl.

7. The compound according to claim 1, wherein R 1 It is a cycloalkyl group.

8. The compound according to claim 7, wherein the cycloalkyl group is cyclopropyl.

9. The compound according to claim 1, wherein R 1 It is a haloalkyl group.

10. The compound according to claim 9, wherein the haloalkyl group is fluoromethyl, difluoromethyl, or trifluoromethyl.

11. The compound according to claim 1, wherein R 1 It is aniline.

12. The compound according to claim 11, wherein the aniline is NH2.

13. The compound according to any one of claims 1 to 12, wherein R 2 It is an alkoxy or -O-haloalkyl group.

14. The compound according to claim 13, wherein R 2 It is an alkoxy group.

15. The compound according to claim 14, wherein the alkoxy group is a methoxy or propoxy group.

16. The compound according to claim 13, wherein R 2 It is an -O-haloalkyl group.

17. The compound of claim 16, wherein the haloalkyl group is a difluoromethyl group.

18. The compound according to any one of claims 1 to 17, wherein R 6 It is OCF3.

19. The compound according to any one of claims 1 to 18, wherein R 7 It is NH2.

20. The compound according to any one of claims 1 to 19, wherein R 3 To be optionally used by one or more R 4 Substituted heterocyclic groups, -NH-L 1 -N(R A R B -NH-L 2 -cycloalkyl or -NH-L 2 - Heterocyclic group, wherein the cycloalkyl group or the heterocyclic group is optionally surrounded by one or more R groups. 5 replace.

21. The compound according to claim 20, wherein R 3 To be optionally used by one or more R 4 Substituted heterocyclic groups.

22. The compound according to claim 21, wherein the heterocyclic group is 1,4-diazacycloheptyl, 1-methyl-1,4-diazacycloheptyl, or 1,4-dimethyl-1,4-di(11-oxoalkyl)-114,414-piperazinyl.

23. The compound of claim 21, wherein the heterocyclic group is pyrrolidinyl, piperidinyl, or piperazineyl, each optionally surrounded by one or more R... 4 Replace, where R 4 C1-C4 alkyl, hydroxyalkyl, -L 1 -N(R A R B -C(O)O-tert-butyl, -L 3 -Aryl or -L 4 - Heterocyclic group.

24. The compound of claim 23, wherein the heterocyclic group is formed by an R 4 Substituted pyrroleyl and R 4 For L 4 - Heterocyclic group, where L 4 The molecule is methylene, and the heterocyclic group is pyrrolidinyl.

25. The compound of claim 23, wherein the heterocyclic group is formed by an R 4 Substituted piperidinyl and R 4 For L 4 - Heterocyclic group, where L 4 The heterocyclic group is a pyrrole alkyl group.

26. The compound of claim 23, wherein the heterocyclic group is piperazine group, and R 4 C1-C4 alkyl, hydroxyalkyl, -L 1 -N(R A R B -C(O)O-tert-butyl; or -L 3 -Aryl.

27. The compound of claim 26, wherein the piperazine group is denoted by an R 4 Replace, and R 4 It is a C1-C4 alkyl group, wherein the C1-C4 alkyl group is methyl or ethyl.

28. The compound of claim 26, wherein the piperazine group is denoted by an R 4 Replace, and R 4 -L 1 -N(R A R B ).

29. The compound of claim 26, wherein the piperazine group is denoted by an R 4 Replace, and R 4 It is -C(O)O-tert-butyl.

30. The compound of claim 26, wherein the piperazine group is affected by two R groups. 4 Group substitution, wherein one R4 group is a C1-C4 alkyl group and the second R4 group is a hydroxyalkyl group or -L 3 -Aryl.

31. The compound according to claim 20, wherein R 3 -NH-L 1 -N(R A R B ).

32. The compound according to claim 31, wherein L 1 It is propylidene and R A and R B Each is a methyl group.

33. The compound according to claim 31, wherein L 1 It is propylidene and R A and R B Each is an ethyl group.

34. The compound according to claim 20, wherein R 3 -NH-L 2 - Heterocyclic group, wherein the heterocyclic group is optionally composed of one or more R 5 replace.

35. The compound according to claim 20, wherein R 3 NH-L 2 -Cycloalkyl, wherein the cycloalkyl group is optionally composed of one or more R 5 replace.

36. The compound according to claim 34, wherein L 2 It is a bond, and the heterocyclic group is optionally bonded by one or more R 5 Substituted pyrrolidinyl groups.

37. The compound of claim 36, wherein the pyrroleyl group is reacted with an R 5 Replace, where R 5 It is ethyl or isopropyl.

38. The compound according to claim 34, wherein L 2 It is a C1-C4 alkylene group, and the heterocyclic group is pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, each optionally surrounded by one or more R groups. 5 replace.

39. The compound according to claim 38, wherein L 2 It is vinylidene or propylene-based.

40. The compound of claim 39, wherein the heterocyclic group is optionally surrounded by one or more R groups. 5 Substituted piperazine group, wherein R 5 It is a methyl group.

41. The compound according to claim 34, wherein L 2 It is a dimethyl vinyl group, and the heterocyclic group is pyrrolidinyl, piperidinyl, or morpholinyl.

42. The compound according to claim 38, wherein L 2 It is a dimethyl vinyl group, and the heterocyclic group is optionally surrounded by one or more R groups. 5 Substituted piperazine group, wherein R 5 It is a methyl group.

43. A compound of formula (I), wherein the compound is: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 , , , , , , , , , , , , , , , , , , , , , , , and .

44. A pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt, and at least one pharmaceutically acceptable excipient or elixir.

45. A method of treating a PLK1-mediated disease or condition in a subject in need, the method comprising administering a therapeutically effective amount of the compound of claims 1 to 43 or a pharmaceutical salt thereof or a pharmaceutical composition of claim 44.

46. ​​The method of claim 45, wherein the PLK1-mediated disease or condition is cancer.

47. The method of claim 46, wherein the PLK1-mediated cancer is melanoma, non-small cell lung cancer (NSCLC), head and neck cancer, esophageal cancer, pharyngeal cancer, breast cancer, liver cancer, endometrial cancer, colorectal cancer, ovarian cancer, pancreatic cancer, or prostate cancer.

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