CN113307797B - Polysubstituted quinazoline compound and application thereof - Google Patents

Abstract

The invention discloses a polysubstituted quinazoline compound and an application thereof, belonging to the field of chemical medicine. The substituted quinazoline compounds of the general formula (I) of the present invention and their pharmaceutically acceptable salts have excellent brain barrier permeability, enhanced metabolic stability, longer metabolic half-life, and are resistant to activation or drug resistance. The mutant form of EGFR shows higher inhibitory activity than wild-type EGFR, which can effectively reduce side effects.

Description

A kind of polysubstituted quinazoline compound and its application

technical field

The invention belongs to the field of chemical medicine, in particular to a polysubstituted quinazoline compound and application thereof.

Background technique

Epidermal growth factor receptor (EGFR) is a member of the erbB receptor family of transmembrane protein tyrosine kinases that, when bound to growth factor ligands such as epidermal growth factor (EGF), can bind to additional Homodimerization of EGFR molecules, or heterodimerization with another family member such as erbB2 (HER2), erbB3 (HER3), or erbB4 (HER4), homodimerization and/or heterodimerization of erbB receptors Dimerization results in the phosphorylation of key tyrosine residues in the cell and in the stimulation of many intracellular signaling pathways involved in cell proliferation and survival. Dysregulation of erbB family signaling promotes proliferation, invasion, metastasis, angiogenesis, and tumor cell survival, and is closely related in human cancers such as lung, head and neck, colon, and breast cancers.

Therefore, the erbB family is an ideal target for anticancer drug development. Specific protein tyrosine kinase inhibitors have attracted much attention as potential anticancer drugs. In 2004, there were reports (Science [2004] No. 304, 1497-1500 and NewEngland Journal of Medicine [2004] No. 350, 2129-2139) about drugs based on this target. Typical representatives of currently marketed EGFR reversible inhibitors include gefitinib and erlotinib, whose structures are as follows, used to inhibit EGFR wild-type and activating mutants (such as exon 19 deletion activation mutation or L858R activating mutation).

Clinical studies have shown that gefitinib and erlotinib have good therapeutic effects on non-small cell lung cancer patients with EGFR exon deletion or L858R point mutation. However, with the emergence of drug resistance, the further clinical application of such inhibitors is limited. Studies have shown that 50% of the drug resistance after gefitinib and erlotinib treatment is secondary to EGFR. Mutation (T790M) associated. Therefore, research to overcome drug resistance caused by T790M mutation is also underway, and irreversible inhibitors have become one of the research directions.

Compared with reversible EGFR inhibitors, irreversible EGFR inhibitors have certain advantages. Irreversible EGFR inhibitors inhibit EGFR for prolonged periods of time, limited only by the normal rate of receptor rebinding. Some studies have found that irreversible EGFR inhibitors can covalently bind to the cysteine residue (Cys797) on EGFR through Michael Addition reaction, so that the irreversible EGFR inhibitors can expand the binding site of ATP, so that they can be used at a certain level. To some extent overcome the drug resistance caused by the T790M mutation (Oncogene [2008], 27:4702-4711). Currently listed or under development irreversible EGFR inhibitors include Afatinib, Neratinib, EKB-569 (Pelitinib), PF00299804 (Dacomitinib) under development, etc. The structures are shown below. However, since such irreversible EGFR inhibitors also have a great inhibitory effect on wild-type EGFR, they will bring greater toxic and side effects, such as diarrhea, nausea, and rash, which limit their clinical application.

A class of pyrimidine compounds is disclosed in the international patent WO2012/061299A1 applied by Avila Therapeutics, a representative compound of which is CO1686 (Rociletinib), with the following structure. It has been reported in the literature that CO1686 can selectively act on EGFR activating mutations and T790M drug resistance mutations, but has a weak inhibitory effect on wild-type EGFR (Cancer Discovery, 2013, 2(12): 1404-1415). However, CO1686 was rejected by the FDA due to the lower-than-expected response rate and side effects of hyperglycemia and QT wave prolongation.

The international patent WO2013/014448A1 applied by AstraZeneca also discloses a series of pyrimidine compounds, of which the representative compound is osimertinib, the structural formula is as follows. Compared with wild-type EGFR, it can activate EGFR T760M mutation and T760M resistance mutation have better inhibitory effect, and the drug has been approved for marketing. The most common adverse reactions (≥25%) of the drug were diarrhea, rash, dry skin, and nail toxicity.

In addition, several generations of inhibitors that have been marketed have shown limited efficacy in the treatment of patients with non-small cell lung cancer with brain metastases, such as: gefitinib, erlotinib, afatinib, osimertinib, etc. Because none of them can effectively cross the blood-brain barrier (BBB) (Journal of Clinical Oncology Official Journal of the American Society of Clinical Oncology, 2006, 24(27): 4517-4520; Neuro-oncology, 2011, 13(12): 1364 -1369). Meanwhile, several reports show that brain metastases from lung cancer appear as an unmet clinical need (Journal of neuro-oncology, 2005, 75(1): 5-14; Journal of Clinical Oncology, 2004, 22(14): 2865-2872 ).

Papilla metastases occur when cancer spreads to the cerebrospinal cord (the layer of tissue that covers the brain and spinal cord). Metastases can spread to the meninges by blood or they can travel from brain metastases carried by cerebrospinal fluid (CSF) that flows through the meninges. If brain tumors enter the CFS and survive, they can travel throughout the central nervous system, leading to neurological problems (Surgical Neurology International, 2013, 4(Suppl 4):S265-S288). The incidence of leptomeningeal metastases is increasing, in part because cancer patients live longer, and because many chemotherapy and molecularly targeted therapies do not reach concentrations in the cerebrospinal fluid sufficient to kill tumor cells.

At the same time, some of the current quinazoline ring drugs have poor efficacy, and it is still necessary to further improve the permeability of the brain barrier, enhance the metabolic stability, improve the pharmacokinetic properties and drug potential.

SUMMARY OF THE INVENTION

The problem to be solved by the invention:

Constructing an unexpectedly excellent brain barrier permeability, enhanced metabolic stability, longer metabolic half-life, and showed higher inhibitory activity against activating or drug-resistant mutant forms of EGFR than wild-type EGFR, A novel polysubstituted quinazoline compound or a pharmaceutically acceptable salt thereof that can effectively reduce side effects.

Solution to problem:

The present invention first provides a compound having the general formula (I) or a pharmaceutically acceptable salt thereof,

in:

When Y is O,

R ¹ is selected from 2-alkenoyl, C4-C8 linear or branched alkyl, C3-C6 cycloalkyl, substituted benzyl or aryl;

R ² is selected from methoxy or H; the substituted group is halogenated C1-C4 alkyl; halogenated includes fluoro, chloro, bromo or iodo;

^{R is} selected from H, F, Cl;

X is selected from NH or O;

Z is selected from

When Y is NH,

R ¹ is selected from 2-alkenoyl, deuterated methyl, C1-C8 straight or branched chain alkyl, C3-C6 cycloalkyl, substituted benzyl or aryl; the substituted group is halogenated C1-C4 alkyl; halo includes fluoro, chloro, bromo or iodo;

R ² is selected from methoxy or H;

^{R is} selected from H, F, Cl;

X is selected from NH or O;

Z is selected from

In one embodiment of the present invention, R ¹ is preferably selected from acryloyl, methyl, deuterated methyl, cyclopropanemethyl, 2-ethylbutyl, 2-methylpentyl, 4-trifluoromethyl benzyl. Further, in some embodiments, R ¹ is preferably selected from acryloyl, deuterated methyl, and cyclopropanemethyl.

In one embodiment of the present invention, Z is preferably selected from

In one embodiment of the present invention, the compound is selected from:

In one embodiment of the present invention, the compound is further preferably the following compound:

In one embodiment of the present invention, the pharmaceutically acceptable salts are inorganic salts or organic salts, and inorganic salts include hydrochloride, hydrobromide, hydroiodide, perchlorate, sulfate , hydrogen sulfate, nitrate, phosphate, acid phosphate; the organic salt is selected from formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, ethyl acetate Diate, Malonate, Succinate, Glutarate, Fumarate, Maleate, Lactate, Malate, Citrate, Tartrate, Mesylate, Ethyl Sulfonates, benzenesulfonates, salicylates, p-toluenesulfonates, ascorbates. Still further, the pharmaceutically acceptable salt is selected from hydrochloride, sulfate, succinate or mesylate.

The present invention also provides the preparation method of the compound described in general formula (I), it comprises:

Reaction scheme one:

As shown in reaction formula 1, intermediate A and intermediate B are nucleophilically substituted to obtain intermediate C; intermediate C is de-Boc protected by trifluoroacetic acid to obtain intermediate D; finally intermediate D and the corresponding aldehyde are reduced Amination yields compound E, wherein R ₁ , R ₂ , X and R ₃ are as defined in general formula (I);

or,

Reaction scheme two:

As shown in Reaction Formula 2, intermediate D and acryloyl chloride are nucleophilically substituted to obtain compound F, wherein the definitions of Y, R ₂ , X and R ₃ are the same as those in general formula (I);

or,

Reaction scheme three:

As shown in reaction formula 3, intermediate G reacts with phenyl chloroformate to obtain intermediate H; intermediate H and (R)-4-Boc-2-methylpiperazine undergo amine transesterification to obtain intermediate I; intermediate I is de-Boc protected by trifluoroacetic acid to obtain intermediate J; finally intermediate J and the corresponding aldehyde are subjected to reductive amination to obtain compound K; wherein, the definitions of R ₁ , R ₂ , X and R ₃ are the same as those of general formula (I) The same definition in;

or,

Reaction scheme four:

As shown in reaction formula 4, intermediate G and diethyl squaraine are transesterified with amine to obtain intermediate L; intermediate L and (R)-4-Boc-2-methylpiperazine are transesterified with amine to obtain intermediate M; intermediate M is de-Boc protected by trifluoroacetic acid to obtain intermediate N; finally intermediate N and formaldehyde are reductively aminated to obtain compound O. Wherein, the definitions of R ₂ and R ₃ are the same as those in the general formula (I).

The third object of the present invention is to provide a pharmaceutical composition comprising the above-mentioned compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

The fourth object of the present invention is to provide the use of the above-mentioned compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating diseases mediated by EGFR-activating or drug-resistant mutants in mammals.

In one embodiment of the present invention, the disease mediated by EGFR-activating or drug-resistant mutants is cancer, specifically including: non-small cell lung cancer or metastatic non-small cell lung cancer.

The fifth object of the present invention is to provide an antitumor drug, which comprises: the above-mentioned compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

In one embodiment of the present invention, the anti-tumor drug may further include the following anti-tumor components:

(i) Antitumor drugs that act on DNA structures;

(ii) antitumor drugs that affect nucleic acid synthesis;

(iii) Antitumor drugs that affect nucleic acid transcription;

(iv) antitumor drugs for tubulin synthesis;

(v) Inhibitors of cell signaling pathways such as epidermal growth factor receptor inhibitors;

(vi) Anti-tumor monoclonal antibody.

Beneficial effects of the present invention:

The present invention provides novel quinazoline inhibitors of activating mutant forms of epidermal growth factor receptors with unexpectedly superior brain barrier penetration properties, making these inhibitors useful for cancers that have metastasized to the CNS The treatment of EGFR, especially brain metastasis and leptomeningeal metastasis, in addition, the inhibitor of the present invention has better pharmacodynamic properties, higher metabolic stability, and shows a stronger response to activating or drug-resistant mutant forms of EGFR than wild-type EGFR. The higher inhibitory activity of EGFR can effectively reduce side effects such as rash and diarrhea.

Detailed ways

The technical solutions of the present invention will be described in detail below with reference to the embodiments.

The term "disease" as used herein refers to any condition or disorder that impairs or interferes with the normal function of cells, organs or tissues.

The term "inhibitor" as used herein refers to a compound or agent having the ability to inhibit the biological function of a targeted protein or polypeptide, eg, by inhibiting the activity or expression of the protein or polypeptide.

The term "anti-neoplastic agent" as used in the present invention refers to any agent useful in the treatment of neoplastic disorders.

The term "pharmaceutically acceptable" as used herein means suitable for use in contact with human and other mammalian tissues without undue toxicity, irritation, allergic reaction, etc., within a reasonable medical range, and with reasonable benefit/ components of the hazard ratio. "Pharmaceutically acceptable salt" refers to any non-toxic salt which, upon administration to a recipient, is capable of providing, directly or indirectly, a compound of the present invention or a prodrug of a compound.

As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a compound or pharmaceutical composition described herein that is sufficient for the intended application, including, but not limited to, treating a disease. In some embodiments, the amount is detected effective to kill or inhibit the growth or spread of cancer cells; the size or number of tumors; or the level of severity, stage, and progression of the cancer. The therapeutically effective amount may vary depending on the intended application, eg, in vitro or in vivo, the state and severity of the disease, the age, weight, or mode of administration of the subject, and the like. The term also applies to doses that will induce a specific response in target cells, eg, reducing cell migration. The specific dose will depend, for example, on the particular compound chosen, the subject species and their age/existing medical condition or risk of medical condition, the route of administration, the severity of the disease, administration in combination with other agents, The time of administration, the tissue to which it is administered, and the delivery device, etc.

In the present invention "administering" or "administering" a compound to an individual refers to providing a compound of the present invention to an individual in need of treatment.

The compounds of the present invention may contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, single enantiomers, individual diastereomers and diastereomeric mixtures. All such isomeric forms of these compounds are expressly included in the present invention. The compounds of the present invention may also exhibit various tautomeric forms, in which case the present invention expressly includes all tautomeric forms of the compounds described herein. All such isomeric forms of such compounds are included in the present invention. All crystalline forms of the compounds described herein are expressly included in the present invention.

<Compound or a pharmaceutically acceptable salt thereof>

The present invention provides a novel quinazoline compound of an activated mutant form of epidermal growth factor receptor or a pharmaceutically acceptable salt thereof, whose structural formula is shown in general formula (I):

Compounds of general formula (I) include pharmaceutically acceptable salts thereof. The pharmaceutically acceptable salts of the present invention are inorganic or organic salts, and inorganic salts include hydrochloride, hydrobromide, hydroiodide, perchlorate, sulfate, hydrogen sulfate, nitrate, phosphoric acid salt, acid phosphate; the organic salt is selected from formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, Succinate, glutarate, fumarate, maleate, lactate, malate, citrate, tartrate, mesylate, ethanesulfonate, benzenesulfonate , salicylate, p-toluenesulfonate, ascorbate. Preferably, from the viewpoint of druggability, the salt of the present invention is hydrochloride, sulfate, succinate or mesylate.

It will be appreciated that certain compounds of formula (I), or pharmaceutically acceptable salts thereof, may exist in solvated forms as well as in unsolvated forms such as, for example, water and forms. It should be understood that the present invention encompasses all such solvate forms possessing inhibitory activity for activating mutant EGFR.

The synthesis of compounds of general formula (I) of the present invention can be accomplished by one of ordinary skill in synthetic chemistry. The documents mentioned in the background of this document are all incorporated by reference in their entirety. The preparation method is described in detail in the examples.

<Pharmaceutical composition>

The present invention provides a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof described in the present invention and a pharmaceutically acceptable carrier, excipient or diluent.

The compounds of the present invention or pharmaceutically acceptable salts thereof can be formulated into solid preparations for oral administration, including, but not limited to, capsules, tablets, pills, powders, granules, and the like. In these solid dosage forms, the compounds of general formula (I) of the present invention are mixed as active ingredient with at least one conventional inert excipient (or carrier), for example with sodium citrate or dicalcium phosphate. Or mixed with the following ingredients: (1) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol and silicic acid, etc.; (2) binders, such as hydroxymethyl cellulose, alginate, gelatin , polyvinyl pyrrolidone, sucrose, gum arabic, etc.; (3) humectants, such as glycerol, etc.; (4) disintegrating agents, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicic acid (5) Solubilizers, such as paraffin, etc.; (6) Absorption accelerators, such as quaternary ammonium compounds, etc.; (7) Wetting agents, such as cetyl alcohol and glycerol monostearate, etc.; (8) ) adsorbents, eg, kaolin, etc.; (9) lubricants, eg, talc, calcium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc., or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

The solid dosage forms such as tablets, dragees, capsules, pills and granules can be coated or microencapsulated with coatings and shell materials such as enteric coatings and other materials well known in the art. They may contain opacifying agents, and the release of the active ingredient in such compositions may be in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric substances and waxes. If desired, the active ingredient may also be in microencapsulated form with one or more of the above-mentioned excipients.

The compounds of the present invention, or pharmaceutically acceptable salts thereof, may be formulated in liquid dosage forms for oral administration, including, but not limited to, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures, and the like. In addition to the compound of general formula (I) or a pharmaceutically acceptable salt thereof as the active ingredient, liquid dosage forms may contain inert diluents conventionally employed in the art, such as water and other solvents, solubilizers and emulsifiers, for example, ethanol , isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil, etc. or a mixture of these substances, etc. Besides these inert diluents, the liquid dosage forms of the present invention may also include conventional adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents, and the like.

Such suspending agents include, for example, ethoxylated stearyl alcohol, polyoxyethylene sorbitol, and sorbitan, microcrystalline cellulose, agar, and the like, or mixtures of these substances.

The compounds of the present invention and their pharmaceutically acceptable salts can be formulated in dosage forms for parenteral injection including, but not limited to, sterile physiologically acceptable aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and Sterile powder for reconstitution into sterile injectable solutions and dispersions. Suitable carriers, diluents, solvents, excipients include water, ethanol, polyols and suitable mixtures thereof.

The compounds of the present invention, or pharmaceutically acceptable salts thereof, may be formulated for topical administration in dosage forms including, for example, ointments, powders, suppositories, drops, propellants, inhalants, and the like. A compound of general formula (I) of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient under sterile conditions and a physiologically acceptable carrier and optional preservatives, buffers, and propellants that may be required if necessary Mix together.

The compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof will be administered to a mammal in a unit dose in the range of 0.01-2000 mg/kg, especially 2.5-1000 mg/kg, especially 5-500 mg/kg, and This should provide an effective dose. However, the daily dose will necessarily vary depending on the host being treated, the particular route of administration, and the severity of the disease being treated. Thus, the optimal dosage can be determined by the practitioner treating any particular patient.

<Use>

The present invention provides a compound of formula (I) as defined above and a pharmaceutically acceptable salt thereof in the manufacture of a compound for the treatment of diseases mediated by EGFR-activating or drug-resistant mutants, especially cancer, in mammals, especially humans. Use in medicine.

In the present invention, the activating mutant form of EGFR, the drug resistant mutant form of EGFR may be, for example, an L858R activating mutant, an Exon19 deletion activating mutant and/or a T790M resistance mutant. Thus, the disease, disorder, disorder or condition mediated by an activating or drug-resistant mutant of EGFR can be, for example, a disease, disorder mediated by an activating mutant of L858R, an activating mutant of deletion of Exon19, and/or a resistance mutant of T790M , disorders or conditions, the present invention is particularly applicable to diseases, disorders, disorders or conditions mediated by EGFR activating mutants such as L858R activating mutants, Exon19 deletion activating mutants. Cancer types that may be susceptible to treatment with a compound of formula (I) or a pharmaceutically acceptable salt thereof include, but are not limited to: ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glial cancer tumor, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin lymphoma, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer , kidney cancer, anaplastic large cell lymphoma, acute myeloid leukemia, multiple myeloma, melanoma and mesothelioma. Preferably, said cancers include non-small cell lung cancer and metastatic non-small cell lung cancer.

For the treatment of cancer according to the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof will be administered to a mammal, more particularly a human.

CNS symptoms such as headache and vomiting that occur in some non-small cell lung cancer patients with CNS metastases (particularly brain metastases and/or leptomeningeal metastases). For these patients, whole brain radiation therapy (WBRT) can be used to improve these symptoms, and when used in combination with WBRT, the compounds of the present invention or pharmaceutically acceptable salts thereof can enhance the antitumor effect of WBRT and further improve CNS symptoms.

Activating mutant forms of EGFR, drug-resistant mutant forms of EGFR activity described herein may be applied as a sole therapy or may involve conventional surgery or radiation therapy in addition to the compounds of the present invention (eg, those described herein). WBRT), may be administered in combination with other pharmaceutically acceptable therapeutic agents, in combination with other antineoplastic agents, and this combination therapy may be achieved by simultaneous, sequential or separate administration of the components of the therapy. The therapeutic agent tumor agents include but are not limited to: anti-tumor drugs that act on the chemical structure of DNA, such as cisplatin, anti-tumor drugs that affect nucleotide synthesis, such as methotrexate, 5-fluorouracil, etc., and anti-tumor drugs that affect nucleic acid transcription. Tumor drugs such as doxorubicin, epirubicin, aclarithromycin, etc., antitumor drugs acting on tubulin synthesis such as paclitaxel, vinorelbine, etc., aromatase inhibitors such as aminoglutamine, letrozole , Ruiningde, etc., cell signaling pathway inhibitors such as epidermal growth factor receptor inhibitors imatinib, gefitinib, erlotinib, afatinib, osimertinib, etc., 6-(4-bromo- 2-Chloro-phenylamino)-7-fluoro-3-methyl-3H-benzimidazole-5-carboxylic acid (2-hydroxy-ethoxy)-amide or a pharmaceutically acceptable salt thereof, 1- [(1S)-1-(imidazo[1,2-a]pyridin-6-yl)ethyl]-6-(1-methyl-1H-pyrazol-4-yl)-1H[1,2 ,3]triazolo[4,5-b]pyrazine or a pharmaceutically acceptable salt thereof. Anti-tumor monoclonal antibodies, such as anti-CTLA-4 antibody, immunosuppressant PD-1, PD-L1, OX40 agonist antibody, etc., the components to be combined can be administered simultaneously or sequentially, in the form of a single preparation or in different preparations. form given. The combination includes not only a combination of one or other active agents of a compound of the present invention, but also a combination of two or more other active agents of a compound of the present invention.

The following examples are intended to illustrate, but not limit, the synthesis of compounds of general formula (I). Temperatures are in degrees Celsius. All evaporations were carried out under reduced pressure unless otherwise stated. If not stated otherwise, reagents were purchased from commercial suppliers and used without further purification. The structures of final products, intermediates, and starting materials are confirmed by standard analytical methods, eg, elemental analysis, spectroscopic characterization, eg, MS, NMR. Abbreviations used are those conventional in the art.

In the specific embodiment, the following intermediate substances are involved, which can be synthesized with reference to the following route process:

Intermediate 6: 4-tert-Butyl-1-4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl(2R)-2-methylpiperin Preparation of oxazine-1,4-dicarboxylate

Step a: Preparation of 4-hydroxy-7-methoxyquinazolin-6-yl acetate (Intermediate 1):

A mixture of 7-methoxyquinazoline-4,6-diol (10.0 g, 52 mmol) and pyridine (8.2 g, 104 mmol) in acetic anhydride (50 mL) was heated to 80° C. for 1 h, and the reaction was completed by TLC detection. The reaction solution was cooled and distilled under reduced pressure, the residue was poured into 200 mL of water, filtered, and the filter cake was vacuum-dried to obtain Intermediate 1 (12.1 g, yield 99%), which could be used in the next reaction without further purification. MS-ESI (m/z): 235.05 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 12.21 (s, 1H), 8.09 (s, 1H), 7.75 (s, 1H ),7.28(s,1H),3.92(s,3H),2.30(s,3H).

Step b: Preparation of 4-chloro-7-methoxyquinazolin-6-yl acetate (Intermediate 2):

To a mixture of Intermediate 1 (12.0 g, 51 mmol) in acetonitrile (100 mL) was added dropwise 1.0 mol/L thionyl chloride (205 mL) under ice bath conditions, followed by 0.5 mL of N,N-dimethylmethane The amide, after dropping, was refluxed at 80°C, and the reaction was detected by TLC. The solvent was evaporated under reduced pressure, the residue was poured into 150 mL of water, and a saturated aqueous sodium bicarbonate solution was slowly added dropwise to the residue under an ice bath, the pH value was adjusted to 8.0-9.0, filtered, the filter cake was washed with water, and the filter cake was vacuum-dried, Intermediate 2 (12.1 g, 93.7% yield) was obtained, which was used in the next reaction without further purification. MS-ESI (m/z): 253.05 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 8.56 (s, 1H), 7.82 (s, 1H), 7.38 (s, 1H ),3.93(s,3H),2.31(s,3H).

Step c: Preparation of 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl acetate (Intermediate 3):

To a suspension of Intermediate 2 (10.0 g, 40 mmol) in acetonitrile (100 mL) was added 2-fluoro-3-chloroaniline (6.3 g, 43 mmol), the mixture was refluxed at 80° C. overnight, and the reaction was completed by TLC detection. The solvent was distilled off under reduced pressure to obtain intermediate 3 (14.3 g, yield 99%), which was used in the next reaction without further purification. MS-ESI (m/z): 362.15 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 11.58 (s, 1H), 8.88 (s, 1H), 8.64 (s, 1H ),7.64(t,J＝7.5Hz,1H),7.54(t,J＝7.4Hz,1H),7.50(s,1H),7.37(t,J＝8.2Hz,1H),4.02(s,3H ),2.40(s,3H).

Step d: Preparation of 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-ol (Intermediate 4):

Anhydrous potassium carbonate (13.7 g, 100 mmol) was added to a mixture of intermediate 3 (14.3 g, 40 mmol) in methanol (150 mL), the mixture was stirred at room temperature overnight, and the reaction was completed by TLC detection. The solvent was evaporated under reduced pressure, the residue was poured into 300 mL of water, filtered, and the filter cake was vacuum-dried to obtain intermediate 4 (9.8 g, yield 77.5%), which could be used in the next reaction without further purification. MS-ESI (m/z): 320.15 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.38 (s, 1H), 8.30 (s, 1H), 7.61 (s, 1H ),7.51(t,J＝7.5Hz,1H),7.3(t,J＝7.4Hz,1H),7.24(t,J＝8.1Hz,1H),7.17(s,1H),3.95(s,3H ).

Step e: 4-tert-Butyl-1-{4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl}(2R)-2-methyl Preparation of piperazine-1,4-dicarboxylate (intermediate 5):

To intermediate 4 (9.8 g, 31 mmol) and tert-butyl(3R)-4-(chlorocarbonyl)-3-methylpiperazine-1-carboxylate (16.1 g, 62 mmol) in N,N-dimethyl Anhydrous potassium carbonate (8.5 g, 62 mmol) was added to the mixture of methylformamide (150 mL), stirred at room temperature overnight, and the reaction was completed by TLC detection. The mixture was poured into 150 mL of water, filtered, and the filter cake was dried in vacuo to give intermediate 5 (15.4 g, 92% yield), which was used in the next reaction without further purification. MS-ESI (m/z): 546.10 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.75 (s, 1H), 8.48 (s, 1H), 8.23 (s, 1H ),7.50(q,J=7.6Hz,2H),7.34(s,1H),7.28(t,J=8.1Hz,1H),4.50～4.20(m,1H),3.98～4.04(m,1H) ,3.95(s,3H),3.82(d,J＝13.7Hz,2H),3.24～3.04(m,2H),2.98～2.82(m,1H),1.44(s,9H),1.23(s,3H ).

Step f: 4-tert-Butyl-1-4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl(2R)-2-methylpiperazine - Preparation of 1,4-dicarboxylate (Intermediate 6):

Intermediate 5 (15.0 g, 2 mmol) was dissolved in a mixed solution of dichloromethane (160 mL) and trifluoroacetic acid (40 mL), reacted at room temperature for 1 h, and the reaction was completed by TLC detection. The solvent was evaporated under reduced pressure, 150 mL of water was added to the residue, the pH was adjusted to 8.0-9.0 with saturated aqueous sodium bicarbonate solution, the layers were separated, the aqueous layer was extracted with 100 mL of dichloromethane, the organic layers were combined, and dried over anhydrous sodium sulfate, The solvent was evaporated under reduced pressure to obtain intermediate 6 (11.4 g, yield 93%), which was used in the next reaction without further purification. MS-ESI (m/z): 446.8 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.74 (s, 1H), 8.47 (s, 1H), 8.22 (s, 1H ),7.50(q,J＝7.9Hz,2H),7.33(s,1H),7.28(t,J＝8.1Hz,1H),4.35～4.10(m,1H),3.95(s,3H),3.85 ～3.60(m,1H), 3.20～3.05(m,1H), 2.99～2.88(m,1H), 2.83～2.73(m,2H), 2.60～2.54(m,1H), 1.32(s,3H) .

Intermediate 13: 4-{[4-((3-Chloro-2-fluorophenyl)amino)-7-methoxyquinazolin-6-yl]carbamoyl}-(R)-3-methyl Preparation of piperazine

Step a: Preparation of 4-chloro-7-methoxy-6-nitroquinazoline (Intermediate 7):

To a mixture of 7-methoxy-6-nitroquinazolin-4-one (4.5 g, 20.3 mmol) and acetonitrile (60 mL) was slowly added dropwise 1.0 mol/L thionyl chloride ( 80 mL), followed by dropwise addition of 2 drops of N,N-dimethylformamide, refluxed at 80° C. for 1 h, and the reaction was completed by TLC detection. The solvent was evaporated under reduced pressure to obtain intermediate 7 (4.8 g, 99% yield), which was used in the next reaction without further purification. MS-ESI(m/z): 234.05[M+H] ⁺ .

Step b: Preparation of 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxy-6-nitroquinazoline (Intermediate 8):

3-Chloro-2-fluoroaniline (3.1 g, 21 mmol) was added dropwise to a mixture of intermediate 7 (4.8 g, 20 mmol) and acetonitrile (60 mL), and the mixture was refluxed at 80° C. for 30 min. TLC detected that the reaction was complete. Cool to room temperature, filter, wash the filter cake with dichloromethane, and dry the filter cake in vacuo to obtain intermediate 8 (6.9 g, yield 99%), which can be used in the next reaction without further purification. MS-ESI (m/z): 349.00 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d6) δ: 11.87 (s, 1H), 9.54 (s, 1H), 8.89 (s, 1H) ,7.64(s,2H),7.54(t,J=6.6Hz,1H),7.37(t,J=8.2Hz,1H),4.12(s,3H).

Step c: Preparation of 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxy-6-aminoquinazoline (Intermediate 9):

Intermediate 8 (6.8 g, 19.5 mmol) was added to a mixed solution of water (40 mL) and ethanol (120 mL), iron powder (5.5 g, 97.5 mmol) and ammonium chloride (7.3 g, 136.5 mmol) were added, and at 90 The temperature was refluxed for 1 h, and the reaction was complete as detected by TLC. Cool to room temperature, adjust the pH to 8-9 with saturated aqueous sodium bicarbonate solution, add 200 mL of ethanol, 100 mL of dichloromethane, 100 mL of ethyl acetate to the system, stir at room temperature for 1 h, filter with diatomaceous earth, wash the filter cake with ethanol, and collect the filtrate , the solvent was evaporated under reduced pressure, 100 mL of water was added to the residue, filtered, and the filter cake was vacuum-dried to obtain intermediate 9 (6.1 g, yield 98%), which could be used in the next reaction without further purification. MS-ESI(m/z): 319.00[M+H] ⁺ ; ¹ H-NMR (400MHz, DMSO-d ₆ )δ: 10.04(s, 1H), 8.43(s, 1H), 7.94～6.78(m ,5H),5.71(s,2H),4.00(s,3H).

Step d: Preparation of phenyl 4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-ylcarbamate (Intermediate 10):

Pyridine (2.7 g, 33.8 mmol) and phenyl chloroformate (3 g, 18.8 mmol) were added to a solution of intermediate 9 (6 g, 18.8 mmol) in N,N-dimethylformamide (20 mL), and reacted at room temperature for 2 h, TLC detection reaction was completed. The reaction solution was added to 100 mL of water, extracted with ethyl acetate, the organic phases were combined, washed with water, dried over anhydrous sodium sulfate, and evaporated to remove the solvent under reduced pressure to obtain Intermediate 10 (6.8 g, yield 82%), the intermediate It was used in the next reaction without further purification. MS-ESI (m/z): 439.00 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.95 (s, 1H), 9.65 (s, 1H), 8.64 (s, 1H ),8.45(s,1H),7.51～7.41(m,4H),7.33～7.22(m,5H),4.03(s,3H).

Step e: 4-{[4-((3-Chloro-2-fluorophenyl)amino)-7-methoxyquinazolin-6-yl]carbamoyl}-(R)-3-methyl Preparation of piperazine-1-carboxylate tert-butyl ester (Intermediate 11):

To a solution of intermediate 10 (6.2 g, 14 mmol) in N,N-dimethylformamide (20 mL) was added (R)-4-Boc-2-methylpiperazine (5.7 g, 28 mmol), and the temperature was raised to 60 The reaction was carried out overnight at °C, and the reaction was completed by TLC detection. The reaction system was added dropwise to 50 mL of ice water, filtered, and the filter cake was vacuum-dried to obtain intermediate 11 (5.7 g, yield 75%), which could be used in the next reaction without further purification. MS-ESI (m/z): 545.10 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.78 (s, 1H), 8.54 (s, 1H), 8.40 (s, 1H ), 8.03(s, 1H), 7.47(q, J=6.7Hz, 2H), 7.25(d, J=8.7Hz, 2H), 4.43～4.30(m, 1H), 4.00(s, 3H), 3.91 ~3.73(m, 3H), 3.10(t, J=12.6Hz, 2H), 1.43(s, 9H), 1.14(d, J=6.7Hz, 3H).

Step f: 4-{[4-((3-Chloro-2-fluorophenyl)amino)-7-methoxyquinazolin-6-yl]carbamoyl}-(R)-3-methyl Preparation of Piperazine (Intermediate 12):

Intermediate 11 (5.5 g, 10.1 mmol) was dissolved in a mixed solution of dichloromethane (40 mL) and trifluoroacetic acid (20 mL), reacted at room temperature for 1 h, and the reaction was completed by TLC detection. The solvent was evaporated under reduced pressure, 60 mL of water was added to the residue, the pH was adjusted to 8.0-9.0 with saturated aqueous sodium bicarbonate solution, the layers were separated, the aqueous layer was extracted with ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate, and then reduced The solvent was evaporated under pressure to give intermediate 12 (4.2 g, 94% yield), which was used in the next reaction without further purification. MS-ESI(m/z): 445.15[M+H] ⁺ .

Intermediate 13: 4-{[4-((3-Chloro-2-fluorophenoxy)-7-methoxyquinazolin-6-yl]carbamoyl}-(R)-3-methyl Preparation of piperazine

The preparation method was referred to the synthesis of intermediate 12 except that 3-chloro-2-fluorophenol was replaced by 3-chloro-2-fluoroaniline.

MS-ESI(m/z): 446.15[M+H] ⁺ . ¹ H-NMR (400MHz, DMSO-d ₆ )δ: 8.78(s, 1H), 8.59(s, 1H), 8.07(s, 1H ),7.60(t,J＝8.0Hz,1H),7.53(t,J＝8.0Hz,1H),7.48(s,1H),7.37(t,J＝8.0Hz,1H),4.33～4.29(m ,1H),4.08(s,3H),3.84～3.78(m,1H),3.20～3.13(m,1H),2.79(d,J＝12.0Hz,1H),2.66(d,J＝8.0Hz, 1H), 2.36～2.29(m, 1H), 2.07(d, J=8.0Hz, 1H), 1.23(s, 3H).

Intermediate 14: Preparation of 4-{[4-((3-Chloro-2-fluorophenyl)amino)quinazolin-6-yl]carbamoyl}-(R)-3-methylpiperazine

The preparation method refers to the synthesis of intermediate 12, except that 6-nitroquinazolin-4-one replaces 7-methoxy-6-nitroquinazolin-4-one.

MS-ESI (m/z): 415.15 [M+H] ⁺ . ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.85 (s, 1H), 8.88 (s, 1H), 8.47 (s, 1H ),8.42(s,1H),7.83(d,J＝8.0Hz,1H),7.73(d,J＝8.0Hz,1H),7.54～7.44(m,2H),7.27(t,J＝8.0Hz ,1H),4.46～4.37(m,1H),3.93(d,J=12.0Hz,2H),3.82～3.7(m,2H),3.14～3.05(m,3H),1.14(d,J=8.0 Hz, 3H).

Intermediate 17: 3-{[4-((3-Chloro-2-fluorophenyl)amino)-7-methoxyquinazolin-6-yl]amino}-(R)-4-(2- Preparation of methylpiperazin-1-yl)cyclobut-3-ene-1,2-dione

Step a: 3-{[(4-((3-Chloro-2-fluorophenyl)amino)-7-methoxyquinazolin-6-yl]amino}-4-ethoxycyclobutan-3 - Preparation of ene-1,2-dione (intermediate 15):

Intermediate 9 (2.5 g, 7.8 mmol) was added to ethanol (30 mL), diethyl squaraine (5.3 g, 31.4 mmol) and zinc trifluoromethanesulfonate (8.6 g, 23.5 mmol) were added, and the mixture was heated at 90° C. The reaction was completed overnight, and the reaction was completed by TLC. Concentrate under reduced pressure, add water to the residue, beat slurry, filter with suction, and dry the filter cake in vacuo to obtain intermediate 15 (1.9 g, yield 54.7%), which can be used in the next reaction without further purification. MS-ESI(m/z): 443.10[M+H] ⁺ .

Step b: 3-{[4-((3-Chloro-2-fluorophenyl)amino)-7-methoxyquinazolin-6-yl]amino}-(R)-4-(4-Boc Preparation of -2-methylpiperazin-1-yl)cyclobut-3-ene-1,2-dione (Intermediate 16):

Intermediate 15 (1.8 g, 3.0 mmol) was added to ethanol (30 mL), (R)-4-Boc-2-methylpiperazine (1.2 g, 6.0 mmol) and zinc triflate (3.3 mmol) were added g, 9.0 mmol), reacted at 90°C overnight. The reaction system was cooled to room temperature, filtered with suction, the filtrate was concentrated under reduced pressure to remove the solvent, and separated and purified by silica gel column chromatography (DCM:MeOH=20:1) to obtain Intermediate 16 (0.65 g, yield 26.8%). MS-ESI(m/z): 597.20[M+H] ⁺ .

Step c: 3-{[4-((3-Chloro-2-fluorophenyl)amino)-7-methoxyquinazolin-6-yl]amino}-(R)-4-(2-methyl) Preparation of ylpiperazin-1-yl)cyclobut-3-ene-1,2-dione (Intermediate 17):

Intermediate 16 (0.6 g, 1.0 mmol) was added to a mixed solution of trifluoroacetic acid (5 mL) and dichloromethane (10 mL), and the reaction was carried out at room temperature for 1 h. TLC detected the end of the reaction. The solvent was evaporated under reduced pressure, 20 mL of water was added to the residue, the pH was adjusted to 8.0-9.0 with saturated aqueous sodium bicarbonate solution, the layers were separated, the aqueous layer was extracted with ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate, and then reduced The solvent was evaporated under pressure to give intermediate 17 (0.46 g, 92.1% yield), which was used in the next reaction without further purification. MS-ESI(m/z): 497.15[M+H] ⁺ .

Intermediate 18: 3-{[4-((3-Chloro-2-fluorophenyl)amino)-quinazolin-6-yl]amino}-(R)-4-(2-methylpiperazine- Preparation of 1-yl)cyclobut-3-ene-1,2-dione

The preparation method refers to the synthesis of intermediate 17, except that 6-nitroquinazolin-4-one replaces 7-methoxy-6-nitroquinazolin-4-one.

Example 1: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl(R)-2-methyl-4-(cyclopropylmethyl ) piperazine-1-carboxylate

Cyclopropylcarbaldehyde (47.24 mg, 0.67 mmol) was added to a solution of intermediate 6 (200 mg, 0.45 mmol) in dichloromethane (15 mL), and the reaction was stirred at room temperature for 0.5 h under nitrogen atmosphere. A mixture of sodium cyanoborohydride (113 mg, 1.8 mmol), acetic acid (2 mL) and methanol (10 mL) was added to the reaction system, stirred at room temperature for 2 h, and the reaction was completed by TLC detection. Water was added to the reaction system, the pH value was adjusted to 8.0-9.0 with saturated aqueous sodium bicarbonate solution, the layers were separated, extracted with dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure, and the mixture was subjected to silica gel column chromatography ( Dichloromethane-methanol, volume ratio 20:1) was separated and purified to obtain the desired product Example (197 mg, yield 88%). MS-ESI (m/z): 500.2 [M+H] ⁺ . ¹ H-NMR (400MHz, DMSO-d6)δ: 9.73(s, 1H), 8.46(s, 1H), 8.21(s, 1H), 7.49(q, J=8.0Hz, 2H), 7.32(s, 1H), 7.27(t, J＝8.0Hz, 1H), 4.32(s, 1H), 3.94(s, 3H), 3.30～3.22(m, 1H), 2.99(d, J＝12.0Hz, 1H), 2.87(d,J＝12.0Hz,1H),2.21(d,J＝8.0Hz,2H),2.17(d,J＝16.0Hz,1H),2.03～1.93(m,1H),1.35(s,3H) ),0.90～0.79(m,1H),0.53～0.48(m,2H),0.13～0.09(m,2H).

Example 2: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl(R)-2-methyl-4-(2-ethylbutane) yl)-piperazine-1-carboxylate

The synthetic method refers to Example 1, wherein 2-ethylbutanal replaces cyclopropylcarbaldehyde.

MS-ESI (m/z): 530.30 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.73 (s, 1H), 8.47 (s, 1H), 8.22 (s, 1H ), 7.56～7.45(m, 2H), 7.33(s, 1H), 7.28(t, J=7.7Hz, 1H), 4.41～4.20(m, 1H), 3.95(s, 3H), 3.86～3.64( m, 1H), 3.31～2.27 (m, 1H), 2.85 (d, J=11.2Hz, 1H), 2.73 (d, J=11.4Hz, 1H), 2.17 (dd, J=12.3, 7.6Hz, 1H) ), 2.10(dd, J＝12.1, 6.6Hz, 2H), 2.00～1.92(m, 1H), 1.48～1.43(m, 1H), 1.40～1.28(m, 7H), 0.86(t, J＝7.4 Hz, 6H).

Example 3: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl(R)-2-methyl-4-(2-methylpentan yl)piperazine-1-carboxylate

The synthetic method refers to Example 1, wherein 2-methylpentanal replaces cyclopropylcarbaldehyde.

MS-ESI (m/z): 530.30 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 9.73 (s, 1H), 8.47 (s, 1H), 8.22 (s, 1H) ,7.54～7.46(m,2H),7.33(s,1H),7.28(t,J=7.7Hz,1H),4.42～4.25(m,1H),3.95(s,3H),3.86～3.72(m ,1H),3.28～3.15(m,1H),2.83(t,J＝12.0Hz,1H),2.73(t,J＝10.6Hz,1H),2.18～1.92(m,4H),1.67(s, 1H), 1.45～1.28(m, 6H), 1.13～1.01(m, 1H), 0.90～0.86(m, 6H).

Example 4: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl(R)-2-methyl-4-[4-(trifluoro Methyl)benzyl]piperazine-1-carboxylate

The synthetic method refers to Example 1, wherein 4-trifluoromethylbenzaldehyde replaces cyclopropylcarbaldehyde.

MS-ESI (m/z): 604.25 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.73 (s, 1H), 8.47 (s, 1H), 8.22 (s, 1H ),7.73(d,J=8.1Hz,2H),7.60(d,J=8.0Hz,2H),7.55～7.50(m,2H),7.34(s,1H),7.28(t,J=8.1Hz ,1H),4.48～4.06(m,2H),3.95(s,3H),3.91～3.77(m,1H),3.68(d,J=14.0Hz,1H),3.58(d,J=14.0Hz, 1H), 2.87(d, J＝11.3Hz, 1H), 2.69(d, J＝11.6Hz, 1H), 2.23(d, J＝10.8Hz, 1H), 2.10(t, J＝11.6Hz, 1H) ,1.38(s,3H).

Example 5: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl(R)-2-methyl-4-acryloylpiperazine-1 - Carboxylate

Triethylamine (91 mg, 0.9 mmol) was slowly added dropwise to a solution (5 mL) of intermediate 6 (200 mg, 0.45 mmol) in dichloromethane under nitrogen atmosphere at 0 °C. After the drop was completed, slowly added dropwise at 0 °C. Acryloyl chloride (41 mg, 0.45 mmol) in dichloromethane (1 mL) solution was dripped, moved to room temperature overnight, and the reaction was complete as detected by TLC. Water was added to the reaction system, extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. MS-ESI (m/z): 500.05 [M+H] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ -d ₆ ) δ: 8.73 (s, 1H), 8.45 (s, 1H), 7.66 (s, 1H), 7.52(s, 1H), 7.32(s, 1H), 7.16(d, J=7.1Hz, 2H), 6.59(s, 1H), 6.39(d, J=16.7Hz, 1H), 5.79( d, J＝10.4Hz, 1H), 4.75～4.43(m, 2H), 4.20～4.06(m, 1H), 3.94(s, 3H), 3.88～3.79(m, 1H), 3.60～3.46(m, 1H), 3.40～3.14(m, 2H), 2.95(s, 1H), 1.33(s, 3H).

Example 6: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl-4-methyl-(R)-2-methylpiperazine- 1-Carboxamide

Paraformaldehyde (20 mg, 0.68 mmol) was added to a solution of intermediate 12 (200 mg, 0.45 mmol) in dichloromethane (10 mL), and the reaction was stirred at room temperature for 0.5 h under nitrogen atmosphere. A mixture of sodium cyanoborohydride (113 mg, 1.8 mmol), acetic acid (2 mL) and methanol (10 mL) was added to the reaction system, stirred at room temperature for 2 h, and the reaction was completed by TLC detection. Water was added to the reaction system, the pH value was adjusted to 8.0-9.0 with saturated aqueous sodium bicarbonate solution, the layers were separated, extracted with dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure, and the mixture was subjected to silica gel column chromatography ( Dichloromethane-methanol, volume ratio 20:1) was separated and purified to obtain the desired product Example (185 mg, yield 90%). MS-ESI (m/z): 459.10 [M+H] ⁺ . ¹ H-NMR (400MHz, DMSO-d6)δ: 9.76(s, 1H), 8.56(s, 1H), 8.40(s, 1H), 7.93(s, 1H), 7.47(q, J=7.7Hz, 2H), 7.26(t, J=9.2Hz, 2H), 4.00(s, 3H), 3.85(d, J=13.2Hz, 1H), 3.20～3.06(m, 2H), 2.79(d, J=10.3 Hz,1H), 2.67(d,J=10.4Hz,1H), 2.36～2.28(m,1H), 2.19(s,3H), 2.10～2.04(m,1H), 1.27(s,3H).

Example 7: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl-4-(cyclopropylmethyl)-(R)-2- Methylpiperazine-1-carboxamide

The synthetic method refers to Example 6, wherein cyclopropylformaldehyde replaces paraformaldehyde.

MS-ESI (m/z): 499.15 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.78 (s, 1H), 8.56 (s, 1H), 8.39 (s, 1H ), 7.93(s, 1H), 7.47(q, J=6.8Hz, 2H), 7.27(t, J=9.2Hz, 2H), 4.37～4.29(m, 1H), 4.00(s, 3H), 3.86 (d,J＝12.7Hz,1H),3.18～3.08(m,1H),2.98(d,J＝11.1Hz,1H),2.86(d,J＝11.2Hz,1H),2.20(d,J＝ 6.2Hz, 2H), 2.13(dd, J＝11.1, 3.8Hz, 1H), 1.97～1.89(m, 1H), 1.27(d, J＝6.6Hz, 3H), 1.17(t, J＝7.1Hz, 1H), 0.57～0.41(m, 2H), 0.12～0.09(m, 2H).

Example 8: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl-4-(2-ethylbutyl)-(R)-2 -Methylpiperazine-1-carboxamide

The synthetic method refers to Example 6, wherein 2-ethylbutyraldehyde replaces paraformaldehyde.

MS-ESI (m/z): 529.25 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.78 (s, 1H), 8.55 (s, 1H), 8.39 (s, 1H ),7.92(s,1H),7.47(q,J＝6.4Hz,2H),7.27(t,J＝9.3Hz,2H),4.34～4.28(m,1H),3.99(s,3H),3.85 (d, J＝12.7Hz, 1H), 3.12 (t, J＝12.2Hz, 1H), 2.82 (d, J＝11.2Hz, 1H), 2.71 (d, J＝11.1Hz, 1H), 2.19～2.09 (m,2H),2.08～2.02(m,1H),1.94～1.86(m,1H),1.28～1.24(m,5H),0.85(d,J=7.4Hz,6H).

Example 9: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl-4-(2-methylpentyl)-(R)-2 -Methylpiperazine-1-carboxamide

The synthetic method refers to Example 6, wherein 2-methylpentanal replaces paraformaldehyde.

MS-ESI (m/z): 529.25 [M+H] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ -d ₆ ) δ: 8.90 (s, 1H), 8.65 (s, 1H), 8.16 (t, J=7.5Hz, 1H), 7.97(s, 1H), 7.45(s, 1H), 7.37(s, 1H), 7.20(t, J=8.0Hz, 1H), 7.13(t, J=8.1Hz, 1H), 4.32～4.23(m, 1H), 4.08(s, 3H), 3.83(d, J=12.7Hz, 1H), 3.44～3.30(m, 1H), 2.91(d, J=11.1Hz, 1H) ),2.77(t,J＝12.9Hz,1H),2.32～2.12(m,5H),1.42(d,J＝7.0Hz,5H),1.18～1.04(m,J＝8.0Hz,2H),0.95 ～0.92(m,3H),0.91～0.88(m,3H).

Example 10: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl-4-[4-(trifluoromethyl)benzyl]-( R)-2-Methylpiperazine-1-carboxamide

The synthetic method refers to Example 6, wherein 4-trifluoromethylbenzaldehyde replaces paraformaldehyde.

MS-ESI (m/z): 603.25 [M+H] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ -d ₆ ) δ: 8.99 (s, 1H), 8.62 (s, 1H), 7.84 (s, 1H), 7.62(d, J＝7.8Hz, 2H), 7.54(s, 1H), 7.51(d, J＝10.1Hz, 2H), 7.33(t, J＝7.4Hz, 1H), 7.18(t, J=8.1Hz, 1H), 4.36～4.24(m, 1H), 4.12(s, 3H), 3.87(d, J=12.7Hz, 1H), 3.74～3.54(m, 2H), 3.46～3.34(m ,1H),3.04～2.92(m,1H),2.75(d,J=11.4Hz,1H),2.44～2.32(m,1H),2.04～1.98(m,1H),1.43(d,J=6.6 Hz, 3H).

Example 11: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl-4-acryloyl-(R)-2-methylpiperazine- 1-Carboxamide

Triethylamine (91 mg, 0.9 mmol) was slowly added dropwise to a solution (5 mL) of intermediate 12 (200 g, 0.45 mmol) in dichloromethane under nitrogen atmosphere at 0 °C. After the drop was completed, slowly added dropwise at 0 °C. Acryloyl chloride (41 mg, 0.45 mmol) in dichloromethane (1 mL) solution was dripped, moved to room temperature overnight, and the reaction was complete as detected by TLC. Water was added to the reaction system, extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, evaporated to remove the solvent under reduced pressure, and separated and purified by silica gel column chromatography (dichloromethane-methanol, volume ratio 20:1) to obtain the objective Product Example (194 mg, 87% yield). MS-ESI (m/z): 499.15 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.76 (s, 1H), 8.53 (s, 1H), 8.40 (s, 1H ), 8.07(s, 1H), 7.48(q, J=7.3, 6.8Hz, 2H), 7.31～7.20(m, 2H), 6.86(d, J=13.3Hz, 1H), 6.18(d, J= 17.0Hz, 1H), 5.74(d, J＝12.5Hz, 1H), 4.30～4.14(m, 1H), 4.00(s, 3H), 3.94(d, J＝12.7Hz, 1H), 3.48～3.38( m, 1H), 3.27～3.04 (m, 3H), 2.92～2.82 (m, 1H), 1.13 (d, J=8.0Hz, 3H).

Example 12: 4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl-4-acryloyl-(R)-2-methylpiperazine- 1-Carboxamide

Anhydrous potassium carbonate (0.62 g, 4.5 mmol) was added to a solution of intermediate 12 (1 g, 2.2 mmol) in acetonitrile (10 mL), and D ₃ -p-toluene was slowly added dropwise to the reaction system at 0° C. under nitrogen atmosphere A solution of sulfonyl methyl ester (0.43 g, 2.3 mmol) in acetonitrile (2 mL) was dripped, moved to room temperature and reacted overnight, and TLC detected that the reaction was complete. Add water to the reaction system, extract with dichloromethane, combine the organic phases, dry over anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and separate and purify by silica gel column chromatography (dichloromethane-methanol, volume ratio 20:1) to obtain the desired product Example (0.78 g, 75% yield). MS-ESI (m/z): 462.15[M+H] ⁺ ; ¹ H-NMR (400MHz, DMSO-d ₆ ) δ 9.79(s, 1H), 8.55(s, 1H), 8.39(s, 1H ), 7.95(s, 1H), 7.47(t, J=7.6Hz, 2H), 7.27(d, J=9.8Hz, 2H), 4.35～4.29(m, 1H), 3.99(s, 3H), 3.87 ～3.81(m, 1H), 3.17～3.08(m, 1H), 2.78(d, J=10.8Hz, 1H), 2.66(d, J=11.2Hz, 1H), 2.06(d, J=8.8Hz, 1H), 1.91～1.82(m, 1H), 1.24(d, J=3.7Hz, 3H).

Example 13::4-(3-Chloro-2-fluorophenoxy)-7-methoxyquinazolin-6-yl-4-methyl-(R)-2-methylpiperazine-1 -Carboxamide

For the synthesis method, refer to Example 6, in which intermediate 13 is substituted for intermediate 12.

MS-ESI (m/z): 460.10 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 8.78 (s, 1H), 8.59 (s, 1H), 8.07 (s, 1H ),7.60(t,J＝7.3Hz,1H),7.53(t,J＝7.5Hz,1H),7.48(s,1H),7.37(t,J＝8.2Hz,1H),4.33～4.29(m ,1H),4.08(s,3H),3.84～3.78(m,1H),3.20～3.13(m,1H),2.79(d,J＝11.0Hz,1H),2.66(d,J＝10.4Hz, 1H), 2.36～2.29(m, 1H), 2.18(s, 3H), 2.07(d, J=8.1Hz, 1H), 1.23(s, 3H).

Example 14: 4-(3-Chloro-2-fluorophenoxy)-7-methoxyquinazolin-6-yl-4-(cyclopropylmethyl)-(R)-2-methyl Piperazine-1-carboxamide

The synthetic method refers to Example 6, wherein intermediate 13 is substituted for intermediate 12, and cyclopropylformaldehyde is substituted for paraformaldehyde.

MS-ESI (m/z): 500.15 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 8.79(s, 1H), 8.59(s, 1H), 8.06(s, 1H ),7.60(t,J＝7.3Hz,1H),7.54(t,J＝7.6Hz,1H),7.48(s,1H),7.40～7.34(m,1H),4.37～4.27(m,1H) ,4.08(s,3H),3.83(d,J＝12.7Hz,1H),3.17(t,J＝11.7Hz,1H),2.99(d,J＝11.3Hz,1H),2.86(d,J＝ 11.2Hz, 1H), 2.20 (d, J=6.6Hz, 2H), 2.13 (d, J=15.2Hz, 1H), 1.99～1.91 (m, 1H), 1.26 (d, J=11.2Hz, 3H) ,0.88～0.79(m,1H),0.51～0.46(m,2H),0.16～0.11(m,2H).

Example 15: 4-(3-Chloro-2-fluorophenoxy)-7-methoxyquinazolin-6-yl-4-(2-ethylbutyl)-(R)-2-methan piperazine-1-carboxamide

The synthetic method refers to Example 6, wherein intermediate 13 is substituted for intermediate 12, and 2-ethylbutyraldehyde is substituted for paraformaldehyde.

MS-ESI (m/z): 530.25 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 8.79 (s, 1H), 8.58 (s, 1H), 8.02 (s, 1H ),7.59(t,J＝7.0Hz,1H),7.55～7.50(m,1H),7.48(s,1H),7.37(t,J＝8.3Hz,1H),4.32～4.26(m,1H) ,4.08(s,3H),3.81(d,J＝12.6Hz,1H),3.20～3.11(m,1H),2.83(d,J＝11.2Hz,1H),2.71(d,J＝11.2Hz, 1H), 2.15(dd, J＝12.2, 7.5Hz, 1H), 2.08(dd, J＝11.5, 7.5Hz, 2H), 1.97～1.87(m, 1H), 1.45(q, J＝6.4Hz, 1H) ),1.38～1.28(m,4H),1.25(d,J=6.8Hz,3H),0.85(t,J=8.0Hz,6H).

Example 16: 4-(3-Chloro-2-fluorophenoxy)-7-methoxyquinazolin-6-yl-4-(2-methylpentyl)-(R)-2-methan piperazine-1-carboxamide

The synthetic method refers to Example 6, wherein intermediate 13 is substituted for intermediate 12, and 2-methylvaleraldehyde is substituted for paraformaldehyde.

MS-ESI (m/z): 530.20 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 8.79 (s, 1H), 8.58 (s, 1H), 8.02 (s, 1H ),7.59(t,J＝8.2Hz,1H),7.53(t,J＝6.9Hz,1H),7.48(s,1H),7.37(t,J＝8.6Hz,1H),4.33～4.27(m ,1H),4.08(s,3H),3.81(d,J＝12.7Hz,1H),3.21～3.11(m,1H),2.86～2.78(m,1H),2.74～2.66(m,1H), 2.14～2.06(m,2H), 2.04～1.91(m,2H), 1.70～1.64(m,1H), 1.43～1.32(m,2H), 1.29～1.19(m,5H), 0.90～0.86(m ,6H).

Example 17: 4-(3-Chloro-2-fluorophenoxy)-7-methoxyquinazolin-6-yl-4-[4-(trifluoromethyl)benzyl]-(R) -2-Methylpiperazine-1-carboxamide

For the synthesis method, refer to Example 6, wherein intermediate 13 replaces intermediate 12, and 4-trifluoromethylbenzaldehyde replaces paraformaldehyde.

MS-ESI (m/z): 604.20 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 8.77 (s, 1H), 8.59 (s, 1H), 8.08 (s, 1H ),7.72(d,J＝8.4Hz,2H),7.59(d,J＝7.9Hz,3H),7.48(s,1H),7.37(t,J＝7.4Hz,1H),4.36～4.29(m ,1H),4.08(s,3H),3.85(d,J＝12.8Hz,1H),3.66(d,J＝14.0Hz,1H),3.56(d,J＝12.0Hz,1H),3.21(t , J＝12.3Hz, 1H), 2.84(d, J＝11.1Hz, 1H), 2.67(d, J＝10.0Hz, 1H), 2.20(d, J＝7.5Hz, 1H), 2.07(t, J =10.8Hz,1H),1.28(d,J=6.6Hz,3H).

Example 18: 4-(3-Chloro-2-fluorophenoxy)-7-methoxyquinazolin-6-yl-4-deuteromethyl-(R)-2-methylpiperazine- 1-Carboxamide

For the synthesis method, refer to Example 12, in which intermediate 14 is substituted for intermediate 13.

MS-ESI (m/z): 463.15 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 8.78(s, 1H), 8.59(s, 1H), 8.05(s, 1H ), 7.60(t, J＝7.6Hz, 1H), 7.53(t, J＝8.0Hz, 1H), 7.48(s, 1H), 7.37(t, J＝8.1Hz, 1H), 4.36–4.27(m ,1H),4.09(s,3H),3.86–3.77(m,1H),3.21–3.12(m,1H),2.82–2.74(m,1H),2.66(d,J=10.7Hz,1H), 2.12–2.04 (m, 1H), 1.92–1.84 (m, 1H), 1.24 (s, 3H).

Example 19::4-(3-Chloro-2-fluorophenoxy)-7-methoxyquinazolin-6-yl-4-acryloyl-(R)-2-methylpiperazine-1 -Carboxamide

For the synthesis method, refer to Example 11, in which Intermediate 13 replaces Intermediate 12.

MS-ESI (m/z): 500.15[M+H] ⁺ ; ¹ H-NMR (400MHz, DMSO-d ₆ )δ: 8.75(s, 1H), 8.60(s, 1H), 8.20(s, 1H ),7.60(t,J＝6.7Hz,1H),7.54(t,J＝7.9Hz,1H),7.49(s,1H),7.37(t,J＝7.5Hz,1H),6.83(dd,J =16.8,10.5Hz,1H),6.18(dd,J=16.6,7.2Hz,1H),5.74(d,J=12.7,1H),4.44–4.37(m,1H),4.09(s,3H), 3.90(d,J＝12.0Hz,1H),3.43(d,J＝12.2Hz,1H),3.14(t,J＝12.1Hz,1H),3.18–3.10(m,1H),3.04(d,J =12.7Hz,1H),2.87(t,J=12.1Hz,1H),1.10(d,J=6.8Hz,3H).

Example 20::4-[(3-Chloro-2-fluorophenyl)amino]-quinazolin-6-yl-4-methyl-(R)-2-methylpiperazine-1-carboxamide

For the synthesis method, refer to Example 6, in which intermediate 14 is substituted for intermediate 12.

MS-ESI (m/z): 429.05 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.83 (s, 1H), 8.80 (s, 1H), 8.48 (s, 1H ), 8.41(s, 1H), 7.83(d, J＝6.8Hz, 1H), 7.72(d, J＝9.0Hz, 1H), 7.49(dt, J＝14.5, 7.5Hz, 2H), 7.27(t , J=8.1Hz, 1H), 4.34 (t, J=5.1Hz, 1H), 3.91 (d, J=13.2Hz, 1H), 3.48–3.40 (m, 1H), 3.17 (d, J=5.3Hz) ,1H),3.13–3.03(m,1H),2.80(d,J=11.2Hz,1H),2.67(d,J=10.7Hz,1H),2.19(s,3H),1.25(d,J= 6.6Hz, 3H).

Example 21: 4-[(3-Chloro-2-fluorophenyl)amino]-quinazolin-6-yl-4-(cyclopropylmethyl)-(R)-2-methylpiperazine- 1-Carboxamide

For the synthesis method, refer to Example 6, wherein intermediate 14 is substituted for intermediate 12, and cyclopropylformaldehyde is substituted for paraformaldehyde. .

MS-ESI (m/z): 469.10 [M+H] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ -d ₆ ) δ: 8.68 (s, 1H), 8.41 (s, 1H), 8.30 (s, 1H), 7.81(d, J＝8.9Hz, 1H), 7.59(d, J＝8.9Hz, 1H), 7.16(q, J＝7.8Hz, 2H), 7.02(s, 1H), 4.45–4.25( m, 1H), 3.91 (d, J=13.0Hz, 1H), 3.45–3.31 (m, 1H), 3.09 (d, J=11.4Hz, 1H), 2.99 (d, J=11.6Hz, 1H), 2.42–2.24(m,4H),1.26(s,3H),0.89(s,1H),0.63-0.54(m,2H),0.15-0.12(m,2H).

Example 22: 4-[(3-Chloro-2-fluorophenyl)amino]-quinazolin-6-yl-4-(2-ethylbutyl)-(R)-2-methylpiperazine -1-Carboxamide

The synthetic method refers to Example 6, wherein intermediate 14 is substituted for intermediate 12, and 2-ethylbutyraldehyde is substituted for paraformaldehyde.

MS-ESI (m/z): 499.30 [M+H] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ -d ₆ ) δ: 8.66 (s, 1H), 8.43 (s, 1H), 8.22 (s, 1H),7.79(d,J=8.9Hz,1H),7.59(d,J=8.7Hz,1H),7.19-7.09(m,2H),7.05(s,1H),4.36-4.26(m,1H) ),3.87(d,J＝12.9Hz,1H),3.31(d,J＝12.8Hz,1H),2.87(d,J＝11.3Hz,1H),2.73(d,J＝11.5Hz,1H), 2.30-2.20(m, 2H), 2.18-2.14(m, 1H), 2.08-2.02(m, 1H), 1.38(s, 3H), 1.35-1.24(m, 5H), 0.88(t, J=7.4 Hz, 6H).

Example 23: 4-[(3-Chloro-2-fluorophenyl)amino]-quinazolin-6-yl-4-(2-methylpentyl)-(R)-2-methylpiperazine -1-Carboxamide

The synthetic method refers to Example 6, wherein intermediate 14 is substituted for intermediate 12, and 2-methylvaleraldehyde is substituted for paraformaldehyde.

MS-ESI (m/z): 499.20 [M+H] ⁺ ; ¹ H-NMR (400 MHz, CDCl ₃ -d ₆ ) δ: 8.66 (s, 1H), 8.46 (s, 1H), 8.22 (s, 1H),7.83(d,J=8.9Hz,1H),7.66(s,1H),7.21～7.09(m,3H),4.42～4.31(m,1H),3.97～3.87(s,1H),3.44 ～3.34(m,1H), 2.99～2.89(m,1H), 2.84～2.75(m,1H), 2.34～2.18(m,4H), 1.38～1.35(m,1H), 1.29～1.22(m, 5H), 1.18～1.00(m, 2H), 0.94～0.88(m, 6H).

Example 24: 4-[(3-Chloro-2-fluorophenyl)amino]-quinazolin-6-yl-4-[4-(trifluoromethyl)benzyl]-(R)-2- Methylpiperazine-1-carboxamide

The synthetic method refers to Example 6, wherein intermediate 14 is substituted for intermediate 12, and 4-trifluoromethylbenzaldehyde is substituted for paraformaldehyde.

MS-ESI (m/z): 573.30 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 8.65 (s, 1H), 8.49 (s, 1H), 8.15 (s, 1H ),7.85(d,J=8.9Hz,1H),7.66(d,J=9.1Hz,1H),7.60(d,J=8.0Hz,2H),7.49(d,J=7.8Hz,2H), 7.16(dq,J＝16.4,8.2Hz,4H),4.39～4.30(m,1H),3.94(d,J＝12.8Hz,1H),3.64(d,J＝13.7Hz,1H),3.52(d , J＝13.6Hz, 1H), 3.36(t, J＝12.3Hz, 1H), 2.90(d, J＝11.2Hz, 2H), 2.70(d, J＝11.4Hz, 2H), 2.31(d, J ＝11.3Hz, 2H), 2.19(t, J＝11.0Hz, 2H), 1.26(s, 3H).

Example 25: 4-[(3-Chloro-2-fluorophenyl)amino]-quinazolin-6-yl-4-acryloyl-(R)-2-methylpiperazine-1-carboxamide

For the synthesis method, refer to Example 11, in which intermediate 14 is substituted for intermediate 12.

MS-ESI (m/z): 469.10 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.87 (s, 1H), 8.93 (s, 1H), 8.48 (s, 1H ),8.42(s,1H),7.84(d,J＝9.1Hz,1H),7.74(d,J＝9.0Hz,1H),7.49(q,J＝8.0Hz,2H),7.27(t,J =8.1Hz,1H),6.96～6.79(m,1H),6.23～6.13(m,1H),5.74(d,J=7.9Hz,1H),4.50～4.42(m,1H),4.00～3.94( m, 1H), 3.43～3.37 (m, 1H), 3.24～3.13 (m, 1H), 3.12～3.02 (m, 1H), 3.00 (d, J=13.9Hz, 1H), 2.82 (t, J= 12.4Hz, 1H), 1.23(s, 3H).

Example 26: 3-{[4-((3-Chloro-2-fluorophenyl)amino)-7-methoxyquinazolin-6-yl]amino}-4-(4-methyl-2 -(R)-Methylpiperazin-1-yl)cyclobut-3-ene-1,2-dione

Paraformaldehyde (18 mg, 0.60 mmol) was added to a solution of intermediate 17 (200 mg, 0.40 mmol) in dichloromethane (10 mL), and the reaction was stirred at room temperature for 0.5 h under nitrogen atmosphere. A mixture of sodium cyanoborohydride (101 mg, 1.6 mmol), acetic acid (2 mL) and methanol (10 mL) was added to the reaction system, stirred at room temperature for 2 h, and the reaction was completed by TLC detection. Water was added to the reaction system, the pH value was adjusted to 8.0-9.0 with saturated aqueous sodium bicarbonate solution, the layers were separated, extracted with dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure, and the mixture was subjected to silica gel column chromatography ( Dichloromethane-methanol, volume ratio 20:1) was separated and purified to obtain the desired product Example (179 mg, yield 87%). MS-ESI (m/z): 511.20 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.60 (s, 1H), 8.42 (s, 1H), 7.89 (s, 1H ),7.49(q,J＝7.0,5.8Hz,2H),7.33～7.18(m,2H),3.99(s,3H),3.67(s,1H),3.44(t,J＝12.4Hz,1H) ,2.74(d,J＝11.3Hz,1H),2.66(d,J＝11.2Hz,1H),2.16(s,3H),1.98(d,J＝12.1Hz,1H),1.90(d,J＝ 11.2Hz, 1H), 1.35(d, J=6.7Hz, 3H).

Example 27: 3-{[4-((3-Chloro-2-fluorophenyl)amino)-quinazolin-6-yl]amino}-4-(4-methyl-2-(R)- Methylpiperazin-1-yl)cyclobut-3-ene-1,2-dione

For the synthetic method, refer to Example 26, in which intermediate 18 is substituted for intermediate 17.

MS-ESI (m/z): 481.20 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.75 (s, 1H), 8.45 (s, 1H), 7.87 (s, 1H ), 7.81–7.65 (m, 3H), 7.55–7.47 (m, 2H), 7.29 (t, J=8.1Hz, 1H), 4.14 (dd, J=5.5, 3.3Hz, 1H), 3.49 (d, J=13.0Hz, 2H), 2.76(d, J=12.0Hz, 1H), 2.67(d, J=4.6Hz, 1H), 2.33(s, 1H), 2.21(d, J=4.2Hz, 1H) ,2.17(s,3H),1.36(d,J＝6.8Hz,3H).

Example 28: 4-[(3,4-Dichloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl(R)-2-methyl-4-acryloylpiperin Azine-1-carboxylate

For the synthesis method, refer to Example 5, wherein 3,4-dichloro-2-fluoroaniline is substituted for 3-chloro-2-fluoroaniline.

MS-ESI (m/z): 534.20 [M+H] ⁺ ; ¹ H NMR (400 MHz, Chloroform-d) δ 8.70 (s, 1H), 8.52 (t, J=8.6 Hz, 1H), 7.65 ( s,1H),7.35(s,3H),6.60(s,1H),6.43–6.39(m,1H),5.83–5.79(m,1H),4.74～4.41(m,2H),4.19～4.08( m, 1H), 3.99(s, 3H), 3.88～3.79(m, 1H), 3.61～3.46(m, 1H), 3.41～3.14(m, 2H), 2.94(s, 1H), 1.62(s, 3H).

Example 29: 4-[(3-Chloro-2,4-difluorophenyl)amino]-7-methoxyquinazolin-6-yl(R)-2-methyl-4-acryloylpiperin Azine-1-carboxylate

For the synthesis method, refer to Example 5, in which 3-chloro-2,4-difluoroaniline is substituted for 3-chloro-2-fluoroaniline.

MS-ESI (m/z): 518.10 [M+H] ⁺ ; ¹ H NMR (400 MHz, Chloroform-d) δ 8.69 (s, 1H), 8.51 (t, J=8.8 Hz, 1H), 7.65 ( s,1H),7.45-7.31(m,3H),6.61(s,1H),6.43-6.38(m,1H),5.83-5.76(m,1H),4.74～4.41(m,2H),4.19～ 4.08(m,1H),3.99(s,3H),3.89～3.79(m,1H),3.63～3.46(m,1H),3.41～3.13(m,2H),2.94(s,1H),1.61( s, 3H).

Example 30: 4-[(3,4-Dichloro-2-fluorophenyl)amino]-quinazolin-6-yl-4-acryloyl-(R)-2-methylpiperazine-1- Carboxamide

For the synthesis method, refer to Example 25, in which 3,4-dichloro-2-fluoroaniline is substituted for 3-chloro-2-fluoroaniline.

MS-ESI (m/z): 503.10 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.87(s, 1H), 8.94(s, 1H), 8.53(s, 1H ),8.45(s,1H),7.88(d,J＝9.1Hz,1H),7.74(d,J＝9.0Hz,1H),7.51(q,J＝8.0Hz,1H),7.27(t,J =8.1Hz,1H),6.96～6.79(m,1H),6.23～6.12(m,1H),5.73(d,J=7.9Hz,1H),4.50～4.42(m,1H),4.00～3.94( m, 1H), 3.43～3.37 (m, 1H), 3.24～3.13 (m, 1H), 3.12～3.02 (m, 1H), 3.00 (d, J=13.9Hz, 1H), 2.82 (t, J= 12.4Hz, 1H), 1.23(s, 3H).

Example 31: 4-[(3-Chloro-2,4-difluorophenyl)amino]-quinazolin-6-yl-4-acryloyl-(R)-2-methylpiperazine-1- Carboxamide

For the synthesis method, refer to Example 25, wherein 3-chloro-2,4-difluoroaniline was replaced by 3-chloro-2-fluoroaniline.

MS-ESI (m/z): 487.20 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.86 (s, 1H), 8.93 (s, 1H), 8.52 (s, 1H ),8.43(s,1H),7.92-7.74(m,2H),7.51(q,J＝8.0Hz,1H),7.27(t,J＝8.1Hz,1H),6.93～6.78(m,1H) ,6.26～6.16(m,1H),5.72(d,J＝7.9Hz,1H),4.51～4.42(m,1H),4.00～3.92(m,1H),3.43～3.35(m,1H),3.26 ～3.15(m, 1H), 3.12～3.02(m, 1H), 3.00(d, J=13.9Hz, 1H), 2.82(t, J=12.4Hz, 1H), 1.23(s, 3H).

Example 32: Biological Activity Assay

NCI-H3255 (L858R) cells were maintained in BEBM medium supplemented with BEGM BulletKit (CC-4175) containing 10% fetal bovine serum (FBS) (Gibco). PC-9 (exon 19 deleted EGFR) cells were maintained in RPMI 1640 (Gibco) containing 10% fetal bovine serum. NCI-H838 (EGFR wild type) cells were maintained in RPMI 1640 (Gibco) containing 10% fetal bovine serum.

All cells were grown at 37°C in a humidified incubator with 5% _CO2 . Measurement of cellular phosphorylation of endogenous p-EGFR in cell lysates was performed according to the protocol described in the Phospho-EGFReceptor (Tyr1068) Sandwich ELISA Kit (Phospho-EGF Receptor (Tyr1068) Sandwich ELISA Kit).

100 μL of cells (32000 cells/well) were seeded in RPMI 1640 + 1% fetal bovine serum in 96-well cell culture plates and incubated overnight at 37°C, 5% CO ₂ . Cells were dosed with compounds serially diluted in 100% DMSO using Tecan. Following the addition of these compounds, the cell plates were incubated for an additional 4 h, (for NCI-H838: rhEGF was added to the cell plates at a final concentration of 100 ng/mL rhEGF to stimulate for 5 minutes), followed by aspirating the medium and adding to each well 110 μL of IP Lysis Buffer (IP Lysis Buffer: Add 1:100 Phosphatase Inhibitor Cocktail 2 & 3, 1:100 Protease Inhibitor Cocktail 2 to IP Lysis Buffer). The plates were placed at 4°C while spinning at 300 rpm for 0.5-1 hour. 100 μL/well of cell lysate was transferred to a coated plate (Cell Signaling Kit) and incubated overnight at 4°C while spinning at 300 rpm. The plates were allowed to go from 4°C to 37°C while spinning at 300 rpm for 1 hour. After aspirating and washing the plates with 1X wash buffer, 100 [mu]L of detection antibody was added to each well. The plate was sealed with tape and incubated at 37°C for 2 hours while spinning at 300 rpm. After aspirating and washing the plates with 1X wash buffer, 100 [mu]L of HRP-labeled secondary antibody (Cell Signaling Kit) was added to each well. The plate was sealed with tape and incubated at 37°C for 1 hour while spinning at 300rpm. After aspirating and washing the plates with 1X wash buffer, 100 [mu]l of TMB substrate (Cell Signaling Kit) was added to each well. The plate was sealed with tape and incubated at 37°C for 30 minutes while spinning at 300rpm. 100 μL of Stop Solution (Cell Signaling Kit) was added to these plates and the absorbance was read at 450 nm over 30 minutes on a SpectraMax M5e microplate reader. The data obtained with each compound is exported into a suitable software package (eg H-BASE) for curve fitting analysis from which _IC50 values are determined.

Aqueous单溶液细胞增殖测定(Promega)来间接测量各孔中的活细胞的数目。这个测定是一种用于通过检测四唑嗡盐向蓝色甲臜衍生物的酶促转化以测量活细胞的代谢活性来测定活细胞的数目的比色方法。添加试剂(20μL)至各孔中，并且使板返回至孵育器中，持续2小时。接着使用Envision板读取器(PerkinElmer)在490nm下测量各孔中的吸光度。通过使用Microsoft XLfit软件或Accelrys Pipeline Pilot，在最佳拟合曲线中相较于DMSO对照，确定使MTS信号减小50％所需的化合物浓度来计算IC₅₀值。The activity of compounds in selectively inhibiting EGFR exon 20 insertion mutations can be assessed using the murine progenitor B cell line Ba/F3 cells that have been transduced with EGFR exon 20 insertions. The expression vector pLVX-IRES puro (Clontech) encoding human EGFR exon 20 insertion A763_Y764insFQEA was transfected into HEK293 cells by the trans lentiviral ORF assembly system (ThermoScientific) to generate virus encoding EGFR exon 20 insertion. Infection by EGFR exon 20 virus was maintained in cells supplemented with 10% fetal bovine serum, 200 μM L-glutamine/200 μg/mL penicillin/200 μg/mL streptomycin (Life Technology) and 10 ng/mL IL-3 (R&D system) in RPMI1640 medium and subsequently selected by puromycin (Life Technology) selection and IL-3 depletion. Ba/F3 cells expressing EGFR exon 20 insertion (designated Ba/F3-EGFR-A763_Y764insFQEA) can proliferate in the absence of IL-3. Determination of the antiproliferative activity of compounds Method: BaF3-EGFR exon 20 cells seeded in 96-well plates ( A763_Y764insFQEA) (2500 cells/well). Plates were incubated for 72 hours in a 37°C incubator under 5% _CO and passed through

Aqueous single solution cell proliferation assay (Promega) was used to indirectly measure the number of viable cells in each well. This assay is a colorimetric method for determining the number of viable cells by detecting the enzymatic conversion of tetrazolium salts to blue formazan derivatives to measure their metabolic activity. Reagents (20 μL) were added to each well and the plate was returned to the incubator for 2 hours. The absorbance in each well was then measured at 490 nm using an Envision plate reader (PerkinElmer). _IC50 values were calculated by determining the concentration of compound required to reduce the MTS signal by 50% compared to the DMSO control in the best fit curve using Microsoft XLfit software or the Accelrys Pipeline Pilot.

The data (nM) for the activity assays for the examples of the invention and the reference compounds are shown in Table 1:

Activity measurement data (nM) of the compounds obtained in the examples of Table 1

where n = the number of repetitions of the experiment.

This shows that Examples 1, 5, 6, 7, 11, 12, 25, 28, 29, 30, 31 have better potency than AZD3759, and Examples 1, 5, 6, 7, 11, 12, 25, 28, 29, 30, 31 are more selective than AZD3759, and have the potential to reduce medical doses and reduce adverse reactions.

Example 33: Blood Brain Barrier Penetration Assay

According to the literature (Journal of Medicinal Chemistry, 2013, 56(1):2-12) Kp, _uuBrain and _Kp , _uuCSF are both the main parameters measured and optimized during CNS drug discovery. Relationship between concentrations of unbound drug in brain and blood Kp _,uuBrain predicts drug effect on metastatic tumors in brain leptomeningeal metastases (LM) caused by metastatic spread of cancer to leptomeninges, metastases Sexual tumors cause central nervous system dysfunction. K _{p,uu CSF} represents the distribution of the drug in the CSF compared to the distribution of the drug in the blood, which drives the drug response during leptomeningeal transfer therapy.

In vitro blood and brain binding assays were performed on HT dialysis plates with semipermeable membranes. Diluted blood (1:1 with DPBS, pH 7.4) and brain homogenate (1:3 with DPBS, pH 7.4) were spiked with 5 μM test compound (in triplicate) and incubated at 37°C with slow rotation. The plates were dialyzed against 150 μL of an equal volume of 100 mM PBS buffer (pH 7.4) for 4 hours. At the end of the incubation, take a 50 μL aliquot from the recipient side and a 5 μL aliquot from the donor chamber. 5 μL of sample was further diluted with 45 μL of blank blood or brain homogenate. Paired samples were matrix matched with buffer or blank blood/brain homogenate and mixed for 2 min, and then precipitated with 150 μL of cold acetonitrile with 100 ng/mL tolbutamide as internal standard. After centrifugation at 4000 rpm for 20 min, the supernatant was diluted with 0.1% aqueous formic acid and analyzed for LC/MS/MS. The unbound fraction (fu) of the test compound in the brain homogenate and diluted blood was calculated by the ratio of the buffer-side reaction to the brain homogenate/blood-side reaction, and using the following equation f _u,bl (f _u,br ) =(1/D)/[(1/fu-1)+1/D)] Calculate the unbound fraction of the test compound in undiluted blood and tissue from the measured fu in homogenate and diluted blood (f _{u, bl} and f _{u, br} ). D is the dilution factor.

The short-term oral absorption (SOA) model is an in vivo screening model used to identify brain penetration of a compound. Six male wistar rats were dosed orally with 2 mg/kg of compound in 1% methylcellulose. Cerebrospinal fluid (CSF) was collected from the cistern at 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, and 7 hours after dosing, and blood samples (>60 μL/time point/each site) were collected via cardiac puncture. EDTA alone coagulates the tube and then immediately dilutes with 3 volumes of water. Brain tissue was harvested and homogenized in 3 volumes of phosphate buffered saline (pH 7.4). All samples were stored at approximately -70°C prior to LC/MS/MS analysis.

Standards were prepared by spiking blank blood, brain homogenate, and artificial CSF from 0.2 ng/mL to 500 ng/mL. Homogenized brain tissue along with blood samples were precipitated by adding 3 volumes of cold acetonitrile containing internal standards (40 ng/mL dexamethasone and 40 ng/mL diclofenac), and 10 μL of CSF was precipitated with 100 μL cold acetonitrile containing internal standards sample. After vortexing for 2 min and centrifuging at 14,000 rpm for 5 min, the supernatant was analyzed by LC/MS/MS. Two sets of standard curves were run at the beginning and end of each batch of analysis from blood samples. A standard curve was made along with the test samples for brain samples and CSF samples.

Total brain levels expressed as brain/blood ratio ( _Kp , brain) were measured by AUC brain/AUC blood in rodents following oral administration. The free fraction of the test compound in the biological matrix was determined by in vitro blood and brain binding assays. Calculate Kp _{,uuBrain and Kp,uuCSF by the following equations: Kp, uuBrain =} AUCBrain/ _AUCBlood ×( _{fu,Brain / Fu.Blood} ) and Kp _,uuCSF = _AUCCSF / (AUC _blood x f _{u. blood} ).

The data determined for the examples of the invention and for AZD3759 are shown in the following table:

Table 2 Examples of the present invention and experimental data of brain barrier permeability of AZD3759

compound K_{p, uu Brain} K_{p，uu CSF} 1 1.65 1.68 5 1.31 1.36 6 1.33 1.35 7 1.52 1.57 11 1.30 1.33 12 1.43 1.46 25 1.33 1.38 28 1.31 1.35 29 1.30 1.35 30 1.32 1.36 AZD3759 1.31 1.35

When compared with AZD3759, Examples 1, 7, and 12 of the present invention have unexpectedly better brain barrier penetration properties, and Examples 5, 6, 11, and 25 demonstrate brain barrier penetration properties comparable to AZD3759, and are more Excellent potency and selectivity.

Example 34: Evaluation of Compound Stability Using Human Liver Microsomes

Determination of Liver Microsomal Enzyme Stability of Example Compounds:

Assay system: The metabolic stability of the compounds of the present invention was tested with 1 mM NADPH using mixed liver microsomes. Samples were analyzed using a mass spectrometer. HRMS was used to determine the peak area response ratio (peak area corresponding to the test compound or control divided by the peak area of the analytical internal standard) without running a standard curve. To detect all possible metabolites, HRMS scans were performed in the appropriate m/z range.

Assay conditions: The assay was performed with one incubation (N=1). Test compounds were incubated at 37°C in buffer containing 0.5 mg/ml liver microsomal protein. Reactions were initiated by addition of cofactors and samples were taken at 0, 2, 4, 8, 16, 24, 36, 48 hours, and positive controls (5 μM testosterone) were incubated in parallel and at 0, 2, 4, 8, 16, 24 , 36, 48 hours sampling.

Assay quality control: The control compound testosterone was performed in parallel to confirm the enzymatic activity of (liver) microsomes. After the final time point, fluorometry was used to confirm the addition of NADPH to the reaction mixture. The T1/2 of the control met an acceptable internal standard.

Analytical method:

Liquid chromatography column: Thermo BDS Hypersil C18 30X2.0mm, 3μm, with guard column M.P., buffer: 25mM formic acid buffer, pH 3.5;

Aqueous phase (A): 90% water, 10% buffer; organic phase (B): 90% acetonitrile, 10% buffer; flow rate: 300 μl/min Autosampler: injection volume 5 μl. See Table 3 for the gradient program.

Table 3 Gradient program

time (minutes) %A %B 0.0 100 0 1.5 0 100 2.0 0 100 2.1 100 0 3.5 100 0

By using human liver microsomes, Examples 1, 7, 12, 25, 28, 30 as described in the present invention exhibited a metabolic half-life greater than 24 hours, significantly greater than that of AZD3759 of 20 hours. Relatively long metabolic half-lives give them the potential to reduce medical doses and extend dosing intervals.

Claims (9)

1. A compound having the general formula (I) or a pharmaceutically acceptable salt thereof,

the following compounds are specifically selected:

2. the compound of claim 1, wherein said pharmaceutically acceptable salt is an inorganic salt or an organic salt; wherein the inorganic salt is selected from any one or more of the following: hydrochloride, hydrobromide, hydroiodide, perchlorate, sulfate, bisulfate, nitrate, phosphate, acid phosphate; the organic salt is selected from any one or more of: formate, acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, succinate, glutarate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, salicylate, p-toluenesulfonate, ascorbate.

3. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

4. Use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating a disease mediated by an EGFR-activating or drug-resistant mutant in a mammal.

5. The use of claim 4, wherein the EGFR-activating or drug-resistant mutant-mediated disease is non-small cell lung cancer.

6. The use of claim 4, wherein the EGFR-activating or drug-resistant mutant-mediated disease is metastatic non-small cell lung cancer.

7. An anti-tumor drug, comprising: a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient, or diluent.

8. The antitumor agent as claimed in claim 7, further comprising any one of the following antitumor components:

(i) antineoplastic drugs acting on the DNA structure;

(ii) antineoplastic agents that affect nucleic acid synthesis;

(iii) anti-tumor drugs that affect nucleic acid transcription;

(iv) tubulin synthesized antineoplastic drugs;

(v) inhibitors of cellular signaling pathways;

(vi) an anti-tumor monoclonal antibody.

9. The antitumor agent as claimed in claim 8, wherein said cell signaling pathway inhibitor is an epidermal growth factor receptor inhibitor.