CN116655599A - Synthesis and application of EGFR allosteric inhibitors - Google Patents

Abstract

The invention relates to synthesis and application of EGFR allosteric inhibitors, and belongs to the field of medicines. The invention provides a compound with a structure shown in a general formula (I)Or a salt thereof, which is capable of selectively inhibiting EGFR and thus useful in the treatment or amelioration of an abnormal cell proliferative disorder.

Description

Synthesis and use of a class of EGFR allosteric inhibitors

Technical Field

The present invention belongs to the field of medicine, and specifically relates to the synthesis and use of a class of EGFR allosteric inhibitors.

Background Art

Epidermal growth factor receptor (EGFR) is a transmembrane protein tyrosine kinase of the ErbB receptor family. When it binds to a growth factor ligand (e.g., epidermal growth factor (EGF)), the receptor can homodimerize with an attached EGFR molecule, or heterodimerize with another family member (e.g., ErbB2 (HER2), ErbB3 (HER3), or ErbB4 (HER4)). The homodimerization and/or heterodimerization of ErbB receptors leads to the phosphorylation of key tyrosine residues in cells and the stimulation of many intracellular signal transduction pathways involved in cell proliferation and survival. Dysregulation of ErbB family signal transduction promotes proliferation, invasion, metastasis, angiogenesis, and survival of tumor cells, and is closely related to human cancers such as lung cancer, head and neck cancer, colon cancer, and breast cancer.

There are currently four generations of EGFR inhibitors. The first generation of EGFR TKIs includes Gefitinib and Erlotinib, which are mainly used to target position 19 deletions and L858R mutations; the second generation includes Afatinib and Dacomitinib, which are mainly used to address the most common T790M-mediated resistance mechanism and covalently bind to the mutated EGFR protein; the third generation of EGFR TKIs is Osimertinib, which has activity against the EGFR TKIs resistance mutation T790M. The fourth generation of EGFR TKIs is mainly divided into two types: ATP competitive inhibitors and allosteric inhibitors, among which the representative drug is the recently launched Mobocertinib. Currently, many allosteric inhibitors have entered the clinic.

In the past decade, molecular targeted therapy represented by epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) has brought great changes to the treatment of cancer, but EGFR-TKI resistance has always been an unresolved problem. For example, EGFR-TKIs are initially effective in patients with non-small cell lung cancer (NSCLC) carrying EGFR mutations, but often evolve into acquired resistance and lose their therapeutic effect, such as the T790M mutation that causes resistance to first/second generation EGFR-TKIs and the C797S mutation that causes resistance to third generation EGFR-TKIs, especially tumors containing L858R/T790M/C797S mutations at the same time, which are helpless against the existing EGFR-TKIs. However, all current EGFR TKIs target the ATP site, and although third-generation irreversible inhibitors (such as osimertinib) can overcome T790M, resistance occurs due to the C797S mutation that has already appeared in treated patients. Cetuximab, an anti-EGFR antibody that blocks receptor dimerization, is ineffective in EGFR-mutant NSCLC because the mutational activation of the kinase is effectively "downstream" of receptor dimerization. Therefore, an alternative strategy is needed to inhibit EGFR. Currently, suitable compounds with alternative mechanisms of action that target mutant EGFR are not available.

As drugs are continuously used in clinical practice, patients often develop varying degrees of drug resistance. Common drug-resistant mutations include EGFR T790M, EGFR C797X, etc., which lead to a decrease in the ability to compete with ATP. Therefore, it is urgent to find new ways to overcome drug resistance. The discovery of allosteric inhibitors has helped researchers find a new binding site, which is located on the back of the ATP pocket. Instead of competing with ATP, the compound binds to the allosteric pocket to stabilize the EGFR protein in an inactive conformation, limiting the activity of the kinase to inhibit the proliferation of lung cancer cells. This is a new strategy for overcoming the drug resistance currently seen at the EGFR target.

In 2019, Black Diamond Therapeutics reported a mutation-selective EGFR allosteric inhibitor, BDTX-189. Studies have shown that EGFR and HER2 ins20 mutations and extracellular domain mutations have inhibitory effects, but their activity needs to be further improved.

Summary of the invention

Problem that the invention aims to solve

The invention provides a novel EGFR allosteric inhibitor, and the structural compound has better pharmacodynamic performance.

Furthermore, the present invention provides a compound having the general formula (I) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition and use thereof.

Solutions for solving problems

The present invention provides a compound having a structure represented by the following general formula (I) or a pharmaceutically acceptable salt thereof:

wherein ring A is selected from a benzene ring, a pyridine ring, a pyrazine ring, _R1 is H, a C1-4 alkyl group, a halogen (F, Cl, Br, I), and _R2 is a halogen (F, Cl, Br, I); or ring A is selected from a benzene ring, a pyrazine ring, _R1 is H, a C1-4 alkyl group, a halogen (F, Cl, Br, I), and _R2 is H;

R ₃ is selected from substituted or unsubstituted C6-10 aryl, C5-12 heteroaryl, and the substituents are arbitrarily selected from 1, 2 or 3 of the following substituents: H, halogen (F, Cl, Br, I);

W is -R ₄ (CH ₂ ) _m -, -O(CH ₂ ) _m -, wherein m is selected from 0, 1, 2, R ₄ is selected from alkenyl, alkynyl; Y ring is selected from C3-6 cycloalkyl, C3-12 heterocyclic group; or -WY is -OR ₅ , R ₅ is selected from C1-4 alkyl;

Z is selected from: H, -NR ₆ , _R6 is selected from H, C1-4 alkyl.

In some embodiments of the present invention, the C3-12 heterocyclic group is selected from a morpholine ring, a piperidine ring, _R7 is selected from C1-4 alkyl.

In some embodiments of the present invention, _R3 is selected from:

In some embodiments of the present invention, W is -R(CH ₂ ) _m -, -O(CH ₂ ) _m -, wherein R is selected from alkynyl, and m is selected from 0, 2;

In the present invention, further preferred compounds of the structure represented by general formula (I) or pharmaceutically acceptable salts thereof are selected from:

The compound of the structure represented by the general formula (I) of the present invention or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is an inorganic salt or an organic salt, the inorganic salt includes hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic salt is selected from acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate, isosulfonate, benzenesulfonate, and salicylate.

Another object of the present invention is to provide a pharmaceutical composition, which contains the compound represented by the above general formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

In another aspect, the present invention provides use of a compound having a structure represented by the above general formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a drug used as an EGFR inhibitor.

In another aspect, the present invention provides use of the compound having the structure represented by the above general formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating cancer.

The cancer mentioned in the present invention can be selected from breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer (such as gastric cancer and colorectal cancer), or lung cancer.

The beneficial effect of the present invention is to design a class of compounds with the biological function of inhibiting EGFR, thereby providing a new means for finding new treatments for cancer, metabolic and immune diseases, cardiovascular diseases, and neurological diseases.

Effects of the invention:

The compounds provided by the present invention having a structure represented by the general formula (I) can be used for therapeutic and/or preventive treatment of patients with cancer, especially non-small cell lung cancer, having EGFR mutations T790M/L858RT790M/L858R/C797S, L858R and/or L858R/C797S, wherein the treatment comprises determining the EGFR activating mutation status of the patient and then administering the compound of formula I or a pharmaceutically acceptable salt thereof as described herein to the patient.

DETAILED DESCRIPTION

The technical solution of the present invention will be described in detail below in conjunction with embodiments.

In the present invention, "C _1-6 alkyl" refers to a saturated linear or branched monovalent hydrocarbon group of 1 to 6 carbon atoms. Examples include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl and 2-methyl-2-propyl.

In the present invention, "heteroaryl" refers to an unsubstituted or substituted stable 5-membered or 6-membered monocyclic aromatic ring system or an unsubstituted or substituted 9-membered or 10-membered benzo-fused heteroaromatic ring system or a bicyclic heteroaromatic ring system, unless otherwise specified, which consists of carbon atoms and 1-4 heteroatoms selected from N, O or S, and wherein the nitrogen or sulfur heteroatom may be selectively oxidized and the nitrogen heteroatom may be selectively quaternized.

The C6-10 aryl group is a monocyclic or bicyclic aromatic hydrocarbon group having 6 to 10 ring atoms, for example, phenyl or naphthyl.

C5-12 heteroaryl refers to a monocyclic or bicyclic aromatic group having 5 to 12 ring atoms, wherein one or more, preferably one, two or three, ring atoms are heteroatoms selected from N, O, S, and the remaining ring atoms are carbon. Representative examples include, but are not limited to, pyrrolyl, thienyl, thiazolyl, imidazolyl, furanyl, methylindolyl, indolyl, isoindolyl, oxazolyl, isoxazolyl, benzothiazolyl, benzoxazolyl, quinolyl, isoquinolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl, tetrazolyl, and the like.

In the present invention, "substituted" means that one or more hydrogen atoms in the group are replaced by the same or different substituents.

"Administration" or "administering" a subject compound in the context of the present invention means providing the subject in need of treatment with a compound of the present invention.

<Compound or Pharmaceutically Acceptable Salt thereof>

The present invention provides a novel oxoisoindole compound or a pharmaceutically acceptable salt thereof as an inhibitor capable of inhibiting epidermal growth factor receptor (EGFR), wherein the structural formula thereof is shown in general formula (I):

<Pharmaceutical Composition>

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

The compounds of the present invention or their pharmaceutically acceptable salts can be formulated into solid preparations for oral administration, including, but not limited to capsules, tablets, pills, powders, granules, etc. In these solid dosage forms, the compounds of the general formula (I) of the present invention are mixed as active ingredients 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; (2) binders, such as hydroxymethylcellulose, alginate, gelatin, polyvinyl pyrrolidone, sucrose, gum arabic, etc.; (3) humectants, such as glycerol, etc.; (4) disintegrators, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates and sodium carbonate, etc.; (5) solubilizers, such as paraffin, etc.; (6) absorption accelerators, such as quaternary ammonium compounds, etc.; (7) wetting agents, such as cetyl alcohol and glyceryl monostearate, etc.; (8) adsorbents, such as kaolin, etc.; (9) lubricants, such as talc, calcium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc., or mixtures thereof. Capsules, tablets and pills may also contain buffers.

The solid dosage forms such as tablets, pills, capsules, pills and granules can be coated or microencapsulated with coating and shell materials such as enteric coatings and other materials known in the art. They can contain opacifiers, and the release of the active ingredient in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active ingredient can also be formed into microcapsules with one or more of the above-mentioned excipients.

The compounds of the present invention or their pharmaceutically acceptable salts can be formulated into liquid dosage forms for oral administration, including, but not limited to, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures, etc. In addition to the compound of formula (I) or its pharmaceutically acceptable salt as an active ingredient, the liquid dosage form may contain inert diluents conventionally used in the art, such as water and other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, in particular cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil, etc. or mixtures of these substances, etc. In addition to these inert diluents, the liquid dosage form of the present invention may also include conventional adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and spices, etc.

The suspending agent includes, for example, ethoxylated stearyl alcohol, polyoxyethylene sorbitol, and sorbitan, microcrystalline cellulose, agar, etc. or a mixture of these substances.

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

The compound of the present invention or its pharmaceutically acceptable salt can be configured into dosage forms for topical administration, including ointments, powders, suppositories, drops, sprays and inhalants, etc. The compound of the present invention of general formula (I) or its pharmaceutically acceptable salt as an active ingredient is mixed with a physiologically acceptable carrier and optional preservatives, buffers, and propellants that may be required under sterile conditions.

The pharmaceutical composition of the present invention comprises a compound of the general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable carrier, excipient, and diluent. When preparing the pharmaceutical composition, the compound of the general formula (I) of the present invention or a pharmaceutically acceptable salt thereof is usually mixed with a pharmaceutically acceptable carrier, excipient, or diluent. The content of the compound of the general formula (I) or a pharmaceutically acceptable salt thereof can be 0.01-1000 mg, for example 0.05-800 mg, 0.1-500 mg, 0.01-300 mg, 0.01-200 mg, 0.05-150 mg, 0.05-50 mg, etc.

<Purpose>

The present invention also provides use of the compound of general formula (I) or a pharmaceutically acceptable salt thereof in preparing a compound for treating cancer in a mammal.

The compounds of general formula (I) or pharmaceutically acceptable salts thereof, the compounds and salts of the present invention can inhibit the activity of epidermal growth factor receptor (EGFR), and are used to treat mammals including humans for therapeutic and/or prophylactic treatment of patients with cancer, especially non-small cell lung cancer, who have EGFR activating mutations. The treatment comprises determining the EGFR activating mutation status of the patient, and then administering the compound of formula I or pharmaceutically acceptable salts thereof as described herein to the patient.

A "therapeutically effective amount" is an amount of a compound of the invention that is effective to produce a biological or medical response in a subject (eg, to reduce or inhibit enzyme or protein activity, or to improve symptoms, alleviate conditions, slow or delay disease progression, or prevent disease).

The cancers mentioned in the present invention include lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin melanoma or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, gallbladder cancer, stomach cancer, colon cancer, breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue carcinoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney cancer or ureteral cancer, renal cell carcinoma, renal pelvis cancer, central nervous system (CNS) cancer, primary central nervous system lymphoma, spinal cord axis cancer, brain stem glioma, pituitary adenoma, etc.

The compounds of the present invention or their pharmaceutically acceptable salts can be administered to mammals, including humans, orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), topically (powders, ointments, drops) or intratumorally.

The compound or pharmaceutically acceptable salt thereof of the present invention can be administered alone, or in combination with other pharmaceutically acceptable therapeutic agents, or in combination with other anti-tumor drugs. This combined therapy can be achieved by using the various components of the treatment simultaneously, sequentially or separately. The therapeutic agent includes, but is not limited to, anti-tumor drugs acting on the chemical structure of DNA, such as cisplatin, anti-tumor drugs affecting nucleotide synthesis, such as methotrexate, 5-fluorouracil, etc., anti-tumor drugs affecting nucleic acid transcription, such as doxorubicin, epirubicin, aclarubicin, etc., anti-tumor drugs acting on microtubule synthesis, such as paclitaxel, vinorelbine, etc., aromatase inhibitors such as aminoglutethimide, letrozole, arinide, etc., cell signaling pathway inhibitors such as ALK inhibitors crizotinib, ceritinib, alectinib, lorlatinib, etc. Anti-tumor monoclonal antibodies, immunosuppressants PD-1, PD-L1, etc., the components to be combined can be administered simultaneously or sequentially, in the form of a single preparation or in the form of different preparations. The combination includes not only a combination of one or other active agents of the compound of the present invention, but also a combination of two or more other active agents of the compound of the present invention.

The present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually based on conventional conditions.

Preparation of 4-chloro-7-methoxyquinazoline-6-carboxylate (Intermediate 1):

4-Chloro-7-methoxyquinazoline-6-carboxylate (80 g, 1.0 eq) was dissolved in thionyl chloride (80 mL), and a catalytic amount of N,N-dimethylformamide (0.2 mL) was added dropwise. The mixture was refluxed at 90°C for 4 h. The reaction was completed by monitoring by TLC. The mixture was cooled to room temperature, and the reaction solution was concentrated under reduced pressure to remove most of the solvent. The residue was slurried with petroleum ether (50 mL), filtered, and the filter cake was washed with petroleum ether and dried to obtain 4-chloro-7-methoxyquinazoline-6-carboxylate (Intermediate 1) (82.5 g, 95.8%) as a gray solid. ¹ H NMR (400 MHz, Chloroform-d) δ: 8.97 (s, 1H), 7.92 (s, 1H), 7.46 (s, 1H), 4.05 (s, 3H), 2.42 (s, 3H).

Preparation of 4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl acetate (Intermediate 2):

4-Chloro-7-methoxyquinazoline-6-carboxylate (Intermediate 1) (20.0 g, 1.0 eq) was dissolved in N-methylpyrrolidone (200 mL) solution, 3-chloro-4-(pyridin-2-ylmethoxy)aniline (18.57 g, 1.0 eq) was added, and the mixture was reacted at room temperature for 2 h. After TLC detection, the reaction was completed, water (400 mL) was added, and the mixture was filtered, the filter cake was washed with water and dried in a vacuum oven (45°C, 24 h) to obtain 4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazoline-6-yl acetate (Intermediate 2) (34.75 g, 97.3%) as a red solid. HRMS (m/z): 451.1626 [M+H] ⁺ .

Preparation of 4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-ol (Intermediate 3):

4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl acetate (Intermediate 2) (34.7 g, 1.0 eq) was dissolved in methanol (400 mL), potassium carbonate (31.9 g, 3.0 eq) was added, and the mixture was stirred at room temperature for 4 h. The reaction was completed after monitoring by TLC. The reaction solution was concentrated under reduced pressure, water was added to the residue, and the mixture was filtered. The filter cake was washed with water and dried in a vacuum oven (45°C, 12 h) to obtain 4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-ol (Intermediate 3) (31.1 g, 98.7%) as a white solid, which was directly used for the next step without purification. HRMS (m/z): 409.1572 [M+H] ⁺ .

Preparation of tert-butyl 4-((4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidine-1-carboxylate (Intermediate 4):

Dissolve 4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-ol (Intermediate 3) (7.7 g, 1.0 eq) in DMF (80 mL), add tert-butyl 4-(toluoyloxy)piperidine-1-carboxylate (8.0 g, 1.2 eq) and potassium carbonate (7.8 g, 3.0 eq), raise the temperature to 80°C, react overnight, monitor the completion of the reaction by TLC, add water (500 mL) to the reaction solution, The solid precipitated and was filtered off with suction. The filter cake was washed with water and dried in a vacuum oven (45°C, 12h) to give a crude product (8.8g). The crude product was purified by column chromatography [DCM:MeOH=20:1 (V/V)] to give tert-butyl 4-((4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidine-1-carboxylate (Intermediate 4) (4.4g, 39.4%) as a white solid. HRMS (m/z): 592.2416 [M+H]+.

Preparation of N-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)-7-methoxy-6-(piperidin-4-yloxy)quinazolin-4-amine (Intermediate 5):

To a reaction flask of tert-butyl 4-((4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidine-1-carboxylate (Intermediate 4) (4.35 g, 1.0 eq) was added a mixed solvent (45 mL) of dichloromethane:trifluoroacetic acid = 4:1 (V/V), and the mixture was reacted at room temperature for 2 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, and a saturated sodium bicarbonate solution was added to adjust the pH to 8-9. Solids precipitated and were filtered off by suction. The filter cake was washed with water and dried in a vacuum oven (45°C, 12 h) to give N-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)-7-methoxy-6-(piperidin-4-yloxy)quinazolin-4-amine (Intermediate 5) (1.73 g, 48.1%) as a brown solid, which was directly used for the next step without purification. HRMS (m/z): 492.1802 [M+H] ⁺ .

Example 1: 1-(4-((4-(3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidin-1-yl)prop-2-en-1-one

Preparation of 2-chloro-1-((3-fluorobenzyl)oxy)-4-nitrobenzene (Intermediate 6):

Under nitrogen protection, 3-chloro-4-fluoroaniline (20g, 1.0eq) was dissolved in N,N-dimethylformamide (200mL) in a three-necked flask, and Cs2CO3 (96.8g, 2.0eq) and (3-fluorophenyl)methanol (17.4g, 1.0eq) were added in sequence, and the temperature was raised to 80°C for overnight reaction. The reaction was completed by TLC monitoring, cooled to room temperature, and ice water (1L) was added to precipitate a white solid, which was filtered, and the filter cake was washed with water and dried to obtain 2-chloro-1-((3-fluorobenzyl)oxy)-4-nitrobenzene (Intermediate 6) (26.8g, 69.1%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.35(d,J＝2.8Hz,1H),8.26(dd,J＝9.2,2.8Hz,1H),7.48(td,J＝9.3,8.5,2.9Hz,2H),7.35-7.31(m,2H),7.21(td,J＝8.7,2.6Hz, 1H),5.41(s,2H).HRMS(m/z):282.2931[M+H] ⁺ .

Preparation of 3-chloro-4-((3-fluorobenzyl)oxy)aniline (Intermediate 7):

3-Chloro-4-((3-fluorobenzyl)oxy)aniline (26.7g, 1.0eq) was dissolved in a mixed solvent of methanol (225mL) and water (75mL), and ammonium chloride (35.6g, 7.0eq) was added. Iron powder (26.5g, 5.0eq) was added three times under heating conditions while stirring, and the temperature was raised to 90°C for reflux reaction, and stirred overnight. The reaction was monitored by TLC to be complete, cooled to room temperature, filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain 3-chloro-4-((3-fluorobenzyl)oxy)aniline (Intermediate 7) (718g, 75.3%) as an off-white solid. ¹ HNMR (400MHz, DMSO-d ₆ ) δ7.48(td,J＝8.0,5.9Hz,1H),7.36-7.24(m,2H),7.25-7.15(m,1H),6.96(d,J＝8.7Hz,1H),6.70(d,J＝2.7Hz,1H),6.52(dd,J＝8. 7,2.7Hz,1H),5.08(s,2H),5.02(s,2H).HRMS(m/z):252.0578[M+H] ⁺ .

Preparation of 4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl acetate (Intermediate 8):

The preparation of Intermediate 8 refers to the synthesis method of Intermediate 2, wherein 3-chloro-4-(pyridin-2-ylmethoxy)aniline is replaced by Intermediate 7. HRMS (m/z): 467.9471 [M+H] ⁺ .

Preparation of 4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-ol (Intermediate 9):

The preparation of Intermediate 9 refers to the synthesis method of Intermediate 3. HRMS (m/z): 426.1516 [M+H] ⁺ .

Preparation of tert-butyl 4-((4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidine-1-carboxylate (Intermediate 10):

The preparation of intermediate 10 refers to the synthesis method of intermediate 4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ9.41(s,1H),8.87(d,J＝1.4Hz,1H),8.72-8.65(m,2H),8.46(s,1H),7.97-7.90(m,2H),7.70(dd,J＝9.0,2.6Hz,1H),7.34(d,J＝9.0Hz,1H),7.22(s,1 H),5.38(s,2H),3.94(s,3H),3.67(dd,J＝13.4,6.9Hz,2H),3.26(s,2H),1.98(d,J＝10.0Hz,2H),1.64(d,J＝12.0Hz,2H),1.42(s,9H).HRMS(m/z):593.2 106[M+H] ⁺ .

Preparation of N-(3-chloro-4-((4-fluorobenzyl)oxy)phenyl)-7-methoxy-6-(piperidin-4-yloxy)quinazolin-4-amine (Intermediate 11):

The preparation of Intermediate 11 refers to the synthesis method of Intermediate 5. HRMS (m/z): 509.2158 [M+H] ⁺ .

Preparation of 1-(4-((4-(3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidin-1-yl)prop-2-en-1-one (Example 1):

Preparation of the compound of Example 1 Refer to the synthesis method of intermediate 6, and replace the base with 2 equivalents of triethylamine. ¹ HNMR (400 MHz, DMSO-d ₆ ) δ9.45 (s, 1H), 8.46 (s, 1H), 7.95 (d, J = 2.9 Hz, 2H), 7.69 (dd, J = 8.9, 2.6 Hz, 1H), 7.48 (td, J = 8.0, 5.9 Hz, 1H), 7.35-7.16 (m, 5H), 6.86 (dd, J = 16.7, 10.5 Hz, 1H), 6.13 (dd, J = 16.7, 2.5 Hz, 1 H),5.69(dd,J＝10.5,2.5Hz,1H),5.26(s,2H),4.82(dq,J＝7.5,3.8Hz,1H),3.94(s,3H),3.87(d,J＝11.2Hz,2H),3.58-3.45(m,2H),2.02(s,2H),1.71( s,2H).HRMS(m/z):563.1870[M+H] ⁺ .

Example 2: 1-(4-((4-(3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidin-1-yl)prop-2-en-1-one

Preparation of 2-((2-chloro-4-nitrophenoxy)methyl)pyrazine (Intermediate 12):

The preparation of intermediate 12 refers to the synthesis method of intermediate 6. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.90 (d, J = 1.4 Hz, 1H), 8.76-8.68 (m, 2H), 8.39 (d, J = 2.8 Hz, 1H), 8.28 (dd, J = 9.1, 2.8 Hz, 1H), 7.56 (d, J = 9.2 Hz, 1H), 5.58 (s, 2H). HRMS (m/z): 266.0032 [M+H] ⁺ .

Preparation of 3-chloro-4-((3-fluorobenzyl)oxy)aniline (Intermediate 13):

The preparation of intermediate 13 refers to the synthesis method of intermediate 7. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.81 (d, J = 1.5 Hz, 1H), 8.69-8.62 (m, 2H), 6.97 (d, J = 8.7 Hz, 1H), 6.66 (d, J = 2.7 Hz, 1H), 6.48 (dd, J = 8.7, 2.7 Hz, 1H), 5.17 (s, 2H), 5.01 (s, 2H). HRMS (m/z): 236.9606 [M+H] ⁺ .

Preparation of 4-((3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl acetate (Intermediate 14):

The preparation of Intermediate 14 refers to the synthesis method of Intermediate 2, wherein 3-chloro-4-(pyridin-2-ylmethoxy)aniline is replaced by Intermediate 13. HRMS (m/z): 452.1489 [M+H] ⁺ .

Preparation of 4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-ol (Intermediate 15):

The preparation of Intermediate 15 refers to the synthesis method of Intermediate 3. HRMS (m/z): 410.1334 [M+H] ⁺ .

Preparation of tert-butyl 4-((4-((3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidine-1-carboxylate (Intermediate 16):

The preparation of intermediate 16 refers to the synthesis method of intermediate 4. ¹ H NMR (400 MHz, DMSO-d ₆ )δ9.41(s,1H),8.87(d,J＝1.4Hz,1H),8.72-8.65(m,2H),8.46(s,1H),7.97-7.90(m,2H),7.70(dd,J＝9.0,2.6Hz,1H),7.34(d,J＝9.0Hz,1H),7.22(s,1 H),5.38(s,2H),3.94(s,3H),3.67(dd,J＝13.4,6.9Hz,2H),3.26(s,2H),1.98(d,J＝10.0Hz,2H),1.64(d,J＝12.0Hz,2H),1.42(s,9H).HRMS(m/z):593.2 106[M+H] ⁺ .

Preparation of N-(3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)-7-methoxy-6-(piperidin-4-yloxy)quinazolin-4-amine (Intermediate 17):

The preparation of Intermediate 17 refers to the synthesis method of Intermediate 5. HRMS (m/z): 493.2154 [M+H] ⁺ .

Preparation of 1-(4-((4-(3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidin-1-yl)prop-2-en-1-one (Example 2):

Example 2 Preparation of the compound Referring to the synthesis method of Intermediate 6, the base was replaced by 2 equivalents of triethylamine. ¹ HNMR (400 MHz, DMSO-d ₆ ) δ 9.46 (s, 1H), 8.87 (s, 1H), 8.71 (d, J = 2.5 Hz, 1H), 8.46 (s, 1H), 7.99-7.93 (m, 2H), 7.71 (dd, J = 9.0, 2.7 Hz, 1H), 7.35 (d, J = 8.9 Hz, 1H), 7.22 (s, 1H), 6.85 (dd, J = 16.7, 10.4 Hz, 1H), 6.27-6.13 (m, 1H),6.14-6.03(m,1H),5.69(dd,J＝10.5,2.5Hz,1H),5.39(s,2H),4.81(tt,J＝7.5,3.6Hz,1H),3.94(s,3H),3.51(s,4H),2.06-2.00(m,2H),1.71(d ,J＝11.9Hz,2H).HRMS(m/z):547.1681[M+H]+.

Example 3: 1-(4-((4-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidin-1-yl)-2-fluoroprop-2-en-1-one

Preparation of 2-fluoroacryloyl chloride (Intermediate 18):

Dissolve methacrylic acid (250 mg, 1.0 eq.) in dichloromethane solution (2.5 mL) in a three-necked flask, add oxalyl chloride (383 mg, 1.1 eq.) in dichloromethane solution (4 mL) dropwise at 0°C under nitrogen protection, and then add a catalytic amount of N,N-dimethylformamide. React at room temperature for 4 hours. After the reaction is completed, concentrate under reduced pressure to obtain 2-fluoroacryloyl chloride as a yellow solid, which is directly used for the next step without purification.

Preparation of 1-(4-((4-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidin-1-yl)-2-fluoroprop-2-en-1-one (Example 3):

The preparation of Example 3 was carried out according to the synthesis method of Intermediate 6, except that acryloyl chloride was replaced by Intermediate 18, and the base was replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.47 (s, 1H), 8.61 (dd, J=5.0, 1.7 Hz, 1H), 8.47 (s, 1H), 7.96 (d, J=2.2 Hz, 2H), 7.89 (td, J=7.7, 1.8 Hz, 1H), 7.68 (dd, J=8.9, 2.6 Hz, 1H), 7.60 (d, J=7.8 Hz, 1H), 7.38 (dd, J=7.5, 4.9 Hz, 1H), 7.28 (d, J=9.0 Hz, 1 H),7.23(s,1H),5.36-5.23(m,4H),4.84(dp,J＝7.2,3.5Hz,1H),3.95(s,3H),3.80(ddd,J＝12.3,7.7,3.9Hz,2H),3.54(ddd,J＝12.5,7.8,3.5Hz,2H),2. 10-2.03(m,2H),1.78(s,2H).HRMS(m/z):564.1910[M+H] ⁺ .

Example 4: 1-(4-((4-(3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidin-1-yl)-2-fluoroprop-2-en-1-one

The preparation of Example 4 was carried out according to the synthesis method of Intermediate 6, except that acryloyl chloride was replaced by Intermediate 18, and the base was replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.55 (s, 1H), 8.47 (s, 1H), 8.03-7.96 (m, 2H), 7.72 (dd, J＝8.9, 2.6 Hz, 1H), 7.48 (td, J＝8.0, 6.0 Hz, 1H), 7.35-7.30 (m, 2H), 7.28-7.22 (m, 2H), 7.19 (ddd, J＝10.3, 8.0, 2.6 Hz, 1H), 5.3 5-5.27(m,2H),5.26(s,2H),4.88(tt,J＝7.3,3.6Hz,1H),3.95(s,3H),3.83(dd,J＝14.4,6.7Hz,2H),3.55(d,J＝10.5Hz,2H),2.11-2.04(m,2H),1.77( s,2H).HRMS(m/z):581.2318[M+H] ⁺ .

Example 5: 1-(4-((4-(3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidin-1-yl)-2-fluoroprop-2-en-1-one

The preparation of Example 5 was carried out according to the synthesis method of Intermediate 6, except that acryloyl chloride was replaced by Intermediate 18, and the base was replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d6) δ9.47 (s, 1H), 8.88 (d, J = 1.4 Hz, 1H), 8.69 (dd, J = 12.8, 2.2 Hz, 2H), 8.47 (s, 1H), 8.00-7.94 (m, 2H), 7.72 (dd, J = 9.0, 2.6 Hz, 1H), 7.35 (d, J = 9.0 Hz, 1H), 7.23 (s, 1H ),5.39(s,2H),5.33-5.15(m,2H),4.84(tt,J＝7.1,3.5Hz,1H),3.95(s,3H),3.80(ddd,J＝12.4,7.6,3.7Hz,2H),3.58-3.49(m,2H),2.06(s,2H),1.78 (s,2H).HRMS(m/z):565.1766[M+H] ⁺ .

Example 6: 1-(4-((4-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidin-1-yl)prop-2-en-1-one

Preparation of tert-butyl (R)-4-(chlorocarbonyl)-3-methylpiperazine-1-carboxylate (Intermediate 19):

Under nitrogen protection at 0°C, bis(trichloromethyl)carbonate (1.0 eq.) and dichloromethane (15 mL) were added to a three-necked flask, followed by pyridine (3.0 eq.) and a dichloromethane (5 mL) solution of tert-butyl (R)-3-methylpiperazine-1-carboxylate (B-1, 1.36 g, 1.0 eq.), and the mixture was reacted overnight at room temperature. The reaction was monitored by TLC, and the solvent was removed by concentration under reduced pressure to obtain tert-butyl (R)-4-(chlorocarbonyl)-3-methylpiperazine-1-carboxylate (Intermediate 19) as a yellow solid. The product was used directly in the next step without purification.

Preparation of 4-(tert-butyl)1-(4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)(R)-2-methylpiperazine-1,4-dicarboxylate (Intermediate 20):

Intermediate 3 (6 g, 1.0 eq) was dissolved in a three-necked flask of N,N-dimethylformamide (60 mL), potassium carbonate (6.1 g, 3.0 eq) was added, and intermediate 19 (0=4.6 g, 1.2 eq) was added under nitrogen protection, and the reaction was allowed to proceed overnight at room temperature. The reaction was completed by monitoring by TLC, and water (400 mL) was added to the reaction solution. Solids precipitated, filtered, and the filter cake was washed with water and dried in a vacuum oven (45°C, 12 h) to obtain 4-(tert-butyl)1-(4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)(R)-2-methylpiperazine-1,4-dicarboxylate (Intermediate 20) (7.9 g, 84.4%) as a brown solid. The product was directly used for the next step without purification. HRMS (m/z): 635.2681 [M+H] ⁺ .

Preparation of 4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-2-methylpiperazine-1-carboxylate (Intermediate 21):

The preparation of intermediate 21 was prepared by referring to the preparation of synthetic method 5. HRMS (m/z): 535.2288 [M+H] ⁺ .

Preparation of 1-(4-((4-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)piperidin-1-yl)prop-2-en-1-one (Example 6):

The preparation of Example 6 was carried out according to the synthesis method of Intermediate 6, except that the base was replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.57 (s, 1H), 8.64-8.53 (m, 2H), 8.31 (s, 1H), 8.06 (d, J=2.6 Hz, 1H), 7.88 (td, J=7.7, 1.9 Hz, 1H), 7.72 (dd, J=9.0, 2.7 Hz, 1H), 7.58 (d, J=7.8 Hz, 1H), 7.41-7.23 (m, 3H), 6.85 (q, J=13.9 Hz, 1 H),6.20(d,J＝16.6Hz,1H),5.78-5.73(m,1H),5.29(s,2H),4.60-4.03(m,3H),4.01(s,1H),3.94(s,3H),3.47(d,J＝13.9Hz,2H),2.99(d,J＝66.5Hz,1 H),1.23(s,3H).HRMS(m/z):588.19[M+H] ⁺ .

Example 7: 4-((3-chloro-4-((5-fluoropyridin-2-yl)methoxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-4-acryloyl-2-methylpiperazine-1-carboxylate

Preparation of 4-(tert-butyl)1-(4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)(R)-2-methylpiperazine-1,4-dicarboxylate (Intermediate 23):

The preparation of intermediate 23 refers to the preparation of intermediate 20. HRMS (m/z): 652.2827 [M+H] ⁺ .

Preparation of 4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-2-methylpiperazine-1-carboxylate (Intermediate 24):

The preparation of intermediate 24 refers to the preparation of intermediate 5. HRMS (m/z): 552.2260 [M+H] +.

Preparation of 4-((3-chloro-4-((5-fluoropyridin-2-yl)methoxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-4-acryloyl-2-methylpiperazine-1-carboxylate (Example 7):

The preparation of Example 7 was carried out according to the synthesis method of Intermediate 6, except that the base was replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.57 (s, 1H), 8.56 (s, 1H), 8.31 (s, 1H), 8.06 (d, J=2.6 Hz, 1H), 7.73 (dd, J=9.0, 2.6 Hz, 1H), 7.47 (td, J=8.0, 6.0 Hz, 1H), 7.35-7.29 (m, 3H), 7.25 (d, J=9.0 Hz, 1H), 7.18 (td, J=8.8, 2.6 Hz, 1H), 6.86 (td, J=16.2, 10.3 Hz, 1H),6.20(dd,J＝16.9,5.9Hz,1H),5.78-5.74(m,1H),5.25(s,2H),4.36(dd,J＝45.6,13.4Hz,2H),3.99(d,J＝13.8Hz,1H),3.94(s,3H),3.47(d,J＝14.0 Hz,2H),3.29-2.86(m,2H),1.22(s,3H).HRMS(m/z):606.2007[M+H] ⁺ .

Example 8: 4-((3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-4-acryloyl-2-methylpiperazine-1-carboxylate

Preparation of 4-(tert-butyl)1-(4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)(R)-2-methylpiperazine-1,4-dicarboxylate (Intermediate 26):

The preparation of intermediate 26 refers to the synthesis method of intermediate 20. HRMS (m/z): 636.2162 [M+H] ⁺ .

Preparation of 4-((3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-2-methylpiperazine-1-carboxylate (Intermediate 27):

The preparation of Intermediate 26 refers to the synthesis method of Intermediate 5. HRMS (m/z): 536.2000 [M+H] ⁺ .

Preparation of 4-((3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-4-acryloyl-2-methylpiperazine-1-carboxylate (Example 8):

The preparation of Example 8 refers to the synthesis method of Intermediate 6, except that the base is replaced by 2 equivalents of triethylamine. ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.58 (s, 1H), 8.64-8.51 (m, 2H), 8.32 (s, 1H), 8.07 (d, J = 2.6Hz, 1H), 7.89 (td, J = 7.7, 1.8Hz, 1H), 7.72 (dd, J = 9.0, 2.6Hz, 1H), 7.5 9(d,J＝7.8Hz,1H),7.41-7.22(m,3H),5.40-5.21(m,4H),4.44(d,J＝71.6Hz, 1H),3.95(s,3H),3.01(s,6H),1.20(d,J＝23.4Hz,3H).HRMS(m/z):590.1628 [M+H] ⁺ .

Example 9: 4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-4-(2-fluoroacryloyl)-2-methylpiperazine-1-carboxylate

The preparation of Example 9 was carried out according to the synthesis method of Example 3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.58 (s, 1H), 8.64-8.51 (m, 2H), 8.32 (s, 1H), 8.07 (d, J=2.6 Hz, 1H), 7.89 (td, J=7.7, 1.8 Hz, 1H), 7.72 (dd, J=9.0, 2.6 Hz, 1H), 7.59 (d, J=7.8 Hz, 1H), 7.41-7.22 (m, 3H), 5.40-5.21 (m, 4H), 4.44 (d, J=71.6 Hz, 1H), 3.95 (s, 3H), 3.01 (s, 6H), 1.23 (s, 3H). HRMS (m/z): 607.1771 [M+H] ⁺ .

Example 10: 4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-4-(2-fluoroacryloyl)-2-methylpiperazine-1-carboxylate

The preparation of Example 10 was carried out according to the synthesis method of Example 3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.57 (s, 1H), 8.56 (s, 1H), 8.31 (s, 1H), 8.06 (d, J=2.6 Hz, 1H), 7.73 (dd, J=9.0, 2.6 Hz, 1H), 7.47 (td, J=8.0, 6.0 Hz, 1H), 7.36-7.14 (m, 5H), 5.41-5.22 (m, 4H), 4.53 (s, 1H), 3.95 (s, 3H), 2.81 (d, J=63.0 Hz, 6H), 1.28-1.18 (m, 3H). HRMS (m/z): 624.1536 [M+H] ⁺ .

Example 11: 4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl (R)-4-(2-fluoroacryloyl)-2-methylpiperazine-1-carboxylate

The preparation of Example 11 was carried out according to the synthesis method of Example 3. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.59 (s, 1H), 8.87 (d, J=1.5 Hz, 1H), 8.72-8.64 (m, 2H), 8.57 (s, 1H), 8.32 (s, 1H), 8.08 (d, J=2.6 Hz, 1H), 7.76 (dd, J=9.0, 2.6 Hz, 1H), 7.35-7.30 (m, 2H), 5.44-5.29 (m, 4H), 4.52 (s, 1H), 3.95 (s, 3H), 3.18 (d, J=5.0 Hz, 6H), 1.23 (s, 3H). HRMS (m/z): 608.1346 [M+H] ⁺ .

Example 12: 1-(4-((4-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidin-1-yl)prop-2-en-1-one

Preparation of tert-butyl 2-methyl-4-(toluoyloxy)piperidine-1-carboxylate (Intermediate 28):

Dissolve tert-butyl 4-hydroxy-2-methylpiperidine-1-carboxylate (2g, 1.0eq) in dichloromethane (20mL), add triethylamine (2.8g, 3.0eq) and DMAP (114mg, 0.1eq) in turn, add p-toluenesulfonyl chloride (1.77g, 1.0eq) in three batches at 0°C, stir overnight at room temperature under nitrogen protection, monitor the reaction by TLC, and concentrate the reaction solution under reduced pressure to obtain a white solid crude product (3.1g). After column chromatography [DCM:MeOH＝10:1(v/v)], tert-butyl 2-methyl-4-(toluoyloxy)piperidine-1-carboxylate (Intermediate 28) (2.5g, 72.8%) was obtained as a light pink solid. HRMS(m/z):392.1988[M+Na] ⁺ .

Preparation of tert-butyl 4-((4-((3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidine-1-carboxylate (Intermediate 29):

The synthesis of Intermediate 29 was carried out according to the synthesis method of Intermediate 4. HRMS (m/z): 606.2643 [M+H] ⁺ .

Preparation of N-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)-7-methoxy-6-((2-methylpiperidin-4-yl)oxy)quinazolin-4-amine (Intermediate 30):

The synthesis of Intermediate 30 was carried out according to the synthesis method of Intermediate 5. HRMS (m/z): 506.2109 [M+H] ⁺ .

Preparation of 1-(4-((4-(3-chloro-4-(pyridin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidin-1-yl)prop-2-en-1-one (Example 12):

The synthesis of Example 12 was carried out according to the synthesis method of Intermediate 6, and the base was replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.43 (s, 1H), 8.45 (s, 1H), 7.95-7.88 (m, 2H), 7.67 (dd, J = 8.9, 2.7 Hz, 1H), 7.48 (td, J = 8.0, 5.9 Hz, 1H), 7.39-7.20 (m, 5H), 6.82 (dd, J = 16.7, 10.5 Hz, 1H), 6.11 (dd, J = 16.6, 2.6 Hz ,1H),5.67(dd,J＝10.4,2.5Hz,1H),5.26(s,2H),5.01(t,J＝3.2Hz,1H),4.57(s,1H),3.95(s,4H),2.14-1.84(m,4H),1.78(s,1H),1.35(d,J＝6.9Hz,3H ).HRMS(m/z):560.2066[M+H] ⁺ .

Example 13: 1-(4-((4-(3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidin-1-yl)prop-2-en-1-one

Preparation of tert-butyl 4-((4-((3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidine-1-carboxylate (Intermediate 31):

The synthesis of Intermediate 31 was carried out according to the synthesis method of Intermediate 4. HRMS (m/z): 623.3984 [M+H] ⁺ .

Preparation of N-(3-chloro-4-((4-fluorobenzyl)oxy)phenyl)-7-methoxy-6-((2-methylpiperidin-4-yl)oxy)quinazolin-4-amine (Intermediate 32)

The synthesis of Intermediate 32 was carried out according to the synthesis method of Intermediate 5. HRMS (m/z): 523.1992 [M+H] ⁺ .

Preparation of 1-(4-((4-(3-chloro-4-((4-fluorobenzyl)oxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidin-1-yl)prop-2-en-1-one (Example 13):

The synthesis of Example 13 was carried out according to the synthesis method of Intermediate 6, and the base was replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.43 (s, 1H), 8.45 (s, 1H), 7.96-7.87 (m, 2H), 7.67 (dd, J = 8.9, 2.7 Hz, 1H), 7.48 (td, J = 8.0, 5.9 Hz, 1H), 7.38-7.21 (m, 5H), 6.82 (dd, J = 16.7, 10.5 Hz, 1H), 6.11 (dd, J = 16.6, 2.6 Hz ,1H),5.67(dd,J＝10.4,2.5Hz,1H),5.26(s,2H),5.01(t,J＝3.2Hz,1H),4.57(s,1H),3.95(s,4H),2.21-1.84(m,4H),1.78(s,1H),1.35(d,J＝6.9Hz,3H ).HRMS(m/z):581.2318[M+H] ⁺ .

Example 14: 1-(4-((4-(3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidin-1-yl)prop-2-en-1-one

Preparation of tert-butyl 4-((4-((3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidine-1-carboxylate (Intermediate 33):

The synthesis of Intermediate 33 was carried out according to the synthesis method of Intermediate 4. HRMS (m/z): 607.1998 [M+H] ⁺ .

Preparation of N-(3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)-7-methoxy-6-((2-methylpiperidin-4-yl)oxy)quinazolin-4-amine (Intermediate 34):

The synthesis of intermediate 34 refers to the synthesis method of intermediate 5. HRMS (m/z): 507.1949 [M+H] +.

Preparation of 1-(4-((4-(3-chloro-4-(pyrazin-2-ylmethoxy)phenyl)amino)-7-methoxyquinazolin-6-yl)oxy)-2-methylpiperidin-1-yl)prop-2-en-1-one (Example 14):

The synthesis of Example 14 was carried out by referring to the synthesis method of Example 1. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.47 (s, 1H), 8.88 (d, J=1.4 Hz, 1H), 8.71 (t, J=2.0 Hz, 1H), 8.67 (d, J=2.6 Hz, 1H), 8.47 (d, J=2.7 Hz, 1H), 7.96 (d, J=2.6 Hz, 1H), 7.91 (s, 1H), 7.71 (dd, J=8.9, 2.6 Hz, 1H), 7.35 (d, J=9.0 Hz, 1H), 7.22 (d, J=5.2 Hz, 1H), 6.87- 6.79(m,1H),6.12(dd,J＝16.7,2.5Hz,1H),5.68(dd,J＝10.4,2.5Hz,1H),5.39(s,2H),5.05-4.99(m,1H),3.95(s,3H),3.93(s,1H),2.51(p,J＝1.9Hz,2 H),2.19-1.84(m,4H),1.35(d,J＝7.0Hz,3H).HRMS(m/z):561.2033[M+H] ⁺ .

Example 15: N-(7-(2-morpholinethoxy)-4-(quinolin-6-ylamino)quinazolin-6-yl)acrylamide

Preparation of 4-(1,4-dioxypyridinyl[4.5]dec-7-en-8-yl)pyridine (Intermediate 35):

The preparation of intermediate 35 refers to the synthesis method of intermediate 1.

Preparation of 7-fluoro-6-nitro-N-(quinolin-6-yl)quinazolin-4-amine (Intermediate 36):

The preparation of Intermediate 36 was carried out according to the synthesis method of Intermediate 2, except that 3-chloro-4-(pyridin-2-ylmethoxy)aniline was replaced by quinolin-6-amine. HRMS (m/z): 336.1103 [M+H] ⁺ .

Preparation of 7-(2-morpholinethoxy)-6-nitro-N-(quinolin-6-yl)quinazolin-4-amine (Intermediate 37):

Intermediate 36 (3 g, 1.0 eq) was dissolved in DMF (30 mL), NaH (0.3 g, 2.0 eq) was added at 0 ° C, and 2-morpholinoethanol (1.76 g, 1.5 eq) was added after stirring for 30 min. The mixture was protected by nitrogen. The mixture was moved to room temperature and reacted overnight. TLC detected that the reaction was complete, and water (180 mL) was added to the reaction solution. Solids precipitated and were filtered. The filter cake was washed with water and dried in a vacuum oven (45 ° C, 24 h) to obtain 7-(2-morpholinoethoxy)-6-nitro-N-(quinolin-6-yl)quinazolin-4-amine (Intermediate 37) (3.71 g, 93%) as a red solid. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.38 (s, 1H), 9.32 (s, 1H), 8.84 (d, J = 4.2Hz, 1H), 8.71 (s, 1H), 8.51 (s, 1H), 8.35 (d, J = 8.3Hz, 1H), 8.17 (d, J = 9.3Hz, 1H), 8.05 ( d,J＝9.2Hz,1H),7.54(d,J＝4.7Hz,2H),4.44(t,J＝5.6Hz,2H),3.58(t,J＝4.7Hz,4H),2.78(t,J＝5.5Hz,2H),2.51(s,4H).HRMS(m/z):447.1833[M+H] ⁺ .

Preparation of 7-(2-morpholinethoxy)-N ⁴ -(quinolin-6-yl)quinazoline-4,6-diamine (Intermediate 38):

The intermediate 37 (3.65 g, 1.0 eq) was dissolved in a solution of methanol: water = (225 mL: 75 mL), and ammonium chloride (3.06 g, 7.0 eq) solid was added. Under heating and stirring at 80°C, iron powder (2.25 g, 5.0 eq) was added in three batches. The mixture was refluxed at 80°C overnight. The reaction was completed after TLC detection. The reaction solution was cooled to room temperature, filtered with diatomaceous earth, and washed with dichloromethane: methanol = 10: 1. The filtrate was concentrated under reduced pressure to obtain a crude product, which was separated and purified by silica gel column chromatography to obtain a brown solid (2.99 g, 87.7%). HRMS (m/z): 417.3158 [M+H] ⁺ .

Preparation of N-(7-(2-morpholinethoxy)-4-(quinolin-6-ylamino)quinazolin-6-yl)acrylamide (Example 15):

The preparation of Example 15 was carried out according to the synthesis method of Intermediate 6, except that the base was replaced by 2 equivalents of triethylamine. ESI-MS m/z: 392.2 [M+H] ⁺ .

Example 16: N-(7-(2-(piperidin-1-yl)ethoxy)-4-(quinolin-6-ylamino)quinazolin-6-yl)acrylamide

Preparation of 6-nitro-7-(2-(piperidin-1-yl)ethoxy)-N-(quinolin-6-yl)quinazolin-4-amine (Intermediate 39):

The preparation of intermediate 39 refers to the synthesis method of intermediate 37. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.39(s,1H),9.32(s,1H),8.84(d,J＝4.3Hz,1H),8.71(s,1H),8.52(s,1H),8.36(d,J＝8.4Hz,1H),8.18(d,J＝9.2Hz,1H),8.05(d,J＝9.1Hz,1H),7.52 (d,J＝9.4Hz,2H),4.41(t,J＝5.7Hz,2H),2.74(t,J＝5.7Hz,2H),2.46(t,J＝5.5Hz,4H),1.49(p,J＝5.6Hz,4H),1.40-1.36(m,2H).HRMS(m/z):445.2557[M+ H] ⁺ .

Preparation of 7-(2-(piperidin-1-yl)ethoxy)-N ⁴ -(quinolin-6-yl)quinazoline-4,6-diamine (Intermediate 40):

The preparation of intermediate 40 refers to the synthesis method of intermediate 38. HRMS (m/z): 415.3186 [M+H] ⁺ .

Preparation of N-(7-(2-(piperidin-1-yl)ethoxy)-4-(quinolin-6-ylamino)quinazolin-6-yl)acrylamide (Example 16):

The preparation of Example 16 was carried out according to the synthesis method of Intermediate 6, except that the base was replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.03 (s, 1H), 9.67 (s, 1H), 8.99 (s, 1H), 8.81 (dd, J = 4.2, 1.7 Hz, 1H), 8.59 (s, 1H), 8.49 (d, J = 2.3 Hz, 1H), 8.32 (dd, J = 8.5, 1.6 Hz, 1H), 8.20 (dd, J = 9.1, 2.4 Hz, 1H), 8.02 (d, J = 9.1 Hz, 1H), 7.50 (dd, J = 8.3, 4.2 Hz, 1H), 7.36 (s, 1 H),6.72(dd,J＝17.0,10.2Hz,1H),6.33(dd,J＝17.1,1.9Hz,1H),5.83(dd,J＝10.1,2.0Hz,1H),4.35(t,J＝5.9Hz,2H),2.82(t,J＝5.8Hz,2H),2.49(s,4H),1. 51(p,J＝5.5Hz,4H),1.39(q,J＝6.0Hz,2H).HRMS(m/z):469.2755[M+H] ⁺ .

Example 17: Preparation of N-(7-(2-morpholinethoxy)-4-(naphthalen-2-ylamino)quinazoline-6-yl)acrylamide

Preparation of 7-fluoro-N-(naphthalen-2-yl)-6-nitroquinazolin-4-amine (Intermediate 41):

The preparation of Intermediate 41 was carried out according to the synthesis method of Intermediate 36, except that 3-chloro-4-(pyridin-2-ylmethoxy)aniline was replaced by naphthalene-2-amine. HRMS (m/z): 335.0960 [M+H] ⁺ .

Preparation of 7-(2-morpholinethoxy)-N-(naphthalen-2-yl)-6-nitroquinazolin-4-amine (Intermediate 42):

The preparation of intermediate 42 refers to the synthesis method of intermediate 37. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.27 (s, 1H), 9.31 (s, 1H), 8.68 (s, 1H), 8.42 (s, 1H), 7.98-7.86 (m, 4H), 7.49 (h, J = 7.2 Hz, 3H), 4.41 (t, J = 5.6 Hz, 2H), 3.58 (t, J = 4.5 Hz, 4H), 2.76 (t, J = 5.6 Hz, 2H), 2.51 (t, J = 4.1 Hz, 4H). HRMS (m/z):

446.0395[M+H] ⁺ .

Preparation of 7-(2-morpholinethoxy)-N4-(naphthalen-2-yl)quinazoline-4,6-diamine (Intermediate 43):

The preparation of intermediate 43 refers to the synthesis method of intermediate 38. HRMS (m/z): 416.2068 [M+H] ⁺ .

Preparation of N-(7-(2-morpholinethoxy)-4-(naphthalen-2-ylamino)quinazolin-6-yl)acrylamide (Example 17):

The preparation of Example 17 refers to the synthesis method of Intermediate 6, and the base is replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.95 (s, 1H), 9.70 (s, 1H), 8.97 (s, 1H), 8.58 (s, 1H), 8.40 (d, J＝2.0 Hz, 1H), 8.00-7.83 (m, 4H), 7.47 (dt, J＝25.5, 7.2 Hz, 2H), 7.35 (s, 1H), 6.72 (dd, J＝17.0, 10.2 Hz, 1H), 6.3 4(dd,J＝17.0,2.0Hz,1H),5.84(dd,J＝10.0,2.0Hz,1H),4.36(t,J＝5.7Hz,2H),3.58(t,J＝4.6Hz,4H),2.84(t,J＝5.7Hz,2H),2.54(d,J＝4.4Hz,4H).HRMS(m/z) :470.1047[M+H] ⁺ .

Example 18: N-(4-(naphthalen-2-ylamino)-7-(2-(piperidin-1-yl)ethoxy)quinazolin-6-yl)acrylamide

Preparation of N-(naphthalen-2-yl)-6-nitro-7-(2-(piperidin-1-yl)ethoxy)quinazolin-4-amine (Intermediate 44):

The preparation of intermediate 44 refers to the synthesis method of intermediate 39. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.28 (s, 1H), 9.30 (s, 1H), 8.68 (s, 1H), 8.42 (s, 1H), 7.98-7.84 (m, 4H), 7.49 (h, J=7.1 Hz, 3H), 4.39 (t, J=5.7 Hz, 2H), 2.72 (t, J=5.6 Hz, 2H), 2.46 (t, J=5.5 Hz, 4H), 1.49 (p, J=5.3 Hz, 4H), 1.38 (q, J=5.6 Hz, 2H). HRMS (m/z): 444.2029 [M+H] ⁺ .

Preparation of 7-(2-morpholinethoxy)-N4-(naphthalen-2-yl)quinazoline-4,6-diamine (Intermediate 45):

The preparation of intermediate 45 refers to the synthesis method of intermediate 38. HRMS (m/z): 417.2522 [M+H] ⁺ .

Preparation of N-(4-(naphthalen-2-ylamino)-7-(2-(piperidin-1-yl)ethoxy)quinazolin-6-yl)acrylamide (Example 18):

The preparation of Example 18 was carried out according to the synthesis method of Intermediate 6, except that the base was replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.94 (s, 1H), 9.74 (s, 1H), 8.99 (s, 1H), 8.56 (s, 1H), 8.39 (d, J=2.0 Hz, 1H), 7.98-7.84 (m, 4H), 7.52-7.40 (m, 2H), 7.34 (s, 1H), 6.74 (dd, J=17.0, 10.3 Hz, 1H), 6.3 3(dd,J＝17.0,2.0Hz,1H),5.83(dd,J＝10.1,2.0Hz,1H),4.34(t,J＝5.8Hz,2H),2.82(s,2H),1.52(p,J＝5.3Hz,4H),1.39(q,J＝5.9Hz,2H).HRMS(m/z):468.1 262[M+H] ⁺ .

Example 19: N-(4-((1-methyl-1H-indol-6-yl)amino)-7-(2-morpholinethoxy)quinazolin-6-yl)acrylamide

Preparation of 1-methyl-6-nitro-1H-indole (Intermediate 47):

Compound 6-nitro-1H-indole (1 g, 1.0 eq) was dissolved in DMF (10 mL), potassium carbonate (2.56 g, 3.0 eq) and iodomethane (1.3 g, 1.5 eq) were added, and the mixture was reacted at room temperature for 4 h. The reaction was completed after monitoring by TLC. Water was added to the reaction solution, and a solid precipitated. The solid was filtered off with suction, and the filtrate was washed with water and dried in a vacuum oven (45 ° C, 12 h) to obtain 1-methyl-6-nitro-1H-indole (intermediate 47) (1.07 g, 98.7%) as a yellow solid.

Preparation of 1-methyl-1H-indol-6-amine (Intermediate 48):

Intermediate 47 (1 g, 1.0 eq) was dissolved in methanol (10 mL), palladium carbon (100 mg, 0.1 eq) was added, and the mixture was reacted overnight at room temperature under hydrogen atmosphere. The reaction was completed by TLC monitoring, and diatomaceous earth was used for filtration. The filtrate was concentrated under reduced pressure to obtain 1-methyl-1H-indole-6-amine (Intermediate 48) (686 mg, 82.7%) as a yellow solid. HRMS (m/z): 147.0398 [M+H] ⁺ .

Preparation of 7-fluoro-N-(1-methyl-1H-indol-6-yl)-6-nitroquinazolin-4-amine (Intermediate 49):

The preparation of intermediate 49 refers to the synthesis method of intermediate 36. HRMS (m/z): 336.1103 [M+H] ⁺ .

Preparation of N-(1-methyl-1H-indol-6-yl)-7-(2-morpholinethoxy)-6-nitroquinazolin-4-amine (Intermediate 50):

The preparation of intermediate 50 refers to the synthesis method of intermediate 37. ¹ H NMR (400MHz, DMSO-d ₆ ) δ10.14(s,1H),9.28(s,1H),8.57(s,1H),7.88(s,1H),7.55(d,J＝8.5Hz,1H),7.47(s,1H),7.39(d,J＝8.5Hz,1H),7.33(d,J＝3.0 Hz,1H),6.42(d,J＝3.0Hz,1H),4.42(t,J＝5.5Hz,2H),3.79(s,3H),3.58(t,J＝4.6Hz,4H),2.78(t,J＝5.5Hz,2H),2.52(s,4H).HRMS(m/z):449.2531[M+H] ⁺ .

Preparation of N ⁴ -(1-methyl-1H-indol-6-yl)-7-(2-morpholinethoxy)quinazoline-4,6-diamine (Intermediate 51):

The preparation of intermediate 51 refers to the synthesis method of intermediate 38. HRMS (m/z): 419.2456 [M+H] ⁺ .

Preparation of N-(4-((1-methyl-1H-indol-6-yl)amino)-7-(2-morpholinethoxy)quinazolin-6-yl)acrylamide (Example 19):

The preparation of Example 19 refers to the synthesis method of Intermediate 6, and the base is replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.95 (s, 1H), 9.70 (s, 1H), 8.97 (s, 1H), 8.58 (s, 1H), 8.40 (d, J＝2.0 Hz, 1H), 8.00-7.83 (m, 4H), 7.47 (dt, J＝25.5, 7.2 Hz, 2H), 7.35 (s, 1H), 6.72 (dd, J＝17.0, 10.2 Hz, 1H), 6.3 4(dd,J＝17.0,2.0Hz,1H),5.84(dd,J＝10.0,2.0Hz,1H),4.36(t,J＝5.7Hz,2H),3.58(t,J＝4.6Hz,4H),2.84(t,J＝5.7Hz,2H),2.54(d,J＝4.4Hz,4H).HRMS: (m/z ):473.1913[M+H] ⁺ .

Example 20: N-(4-((1-methyl-1H-indol-6-yl)amino)-7-(2-(piperidin-1-yl)ethoxy)quinazolin-6-yl)acrylamide

Preparation of N-(1-methyl-1H-indol-6-yl)-6-nitro-7-(2-(piperidin-1-yl)ethoxy)quinazolin-4-amine (Intermediate 52):

The preparation of intermediate 52 refers to the synthesis method of intermediate 39. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.14(s,1H),9.28(s,1H),8.58(s,1H),7.89(d,J＝1.8Hz,1H),7.55(d,J＝8.5Hz,1H),7.46(s,1H),7.40(dd,J＝8.5,1.8Hz,1H),7.33(d,J＝3.1Hz,1H) ,6.42(d,J＝3.1Hz,1H),4.39(t,J＝5.6Hz,2H),2.73(t,J＝5.6Hz,2H),2.46(t,J＝5.4Hz,4H),1.49(p,J＝5.5Hz,4H),1.38(q,J＝6.8,6.2Hz,2H).HRMS(m/z):4 47.2029[M+H] ⁺ .

Preparation of N ⁴ -(1-methyl-1H-indol-6-yl)-7-(2-morpholinethoxy)quinazoline-4,6-diamine (Intermediate 53):

The preparation of intermediate 53 refers to the synthesis method of intermediate 38. HRMS (m/z): 419.2456 [M+H] ⁺ .

Preparation of N-(4-((1-methyl-1H-indol-6-yl)amino)-7-(2-(piperidin-1-yl)ethoxy)quinazolin-6-yl)acrylamide (Example 20):

The preparation of Example 20 refers to the synthesis method of Intermediate 6, and the base is replaced by 2 equivalents of triethylamine. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ9.72 (s, 1H), 9.64 (s, 1H), 8.92 (s, 1H), 8.46 (s, 1H), 7.87 (d, J = 1.6 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.40 (dd, J = 8.5, 1.9 Hz, 1H), 7.29 (d, J = 3.1 Hz, 2H), 6.70 (dd, J = 17.0, 10.2 Hz, 1H), 6.40 (d, J = 3.0 Hz, 1H), 6.3 2(dd,J＝16.9,2.0Hz,1H),5.82(dd,J＝10.3,2.0Hz,1H),4.31(t,J＝5.9Hz,2H),3.78(s,3H),2.80(t,J＝5.8Hz,2H),2.48(t,J＝4.3Hz,4H),1.49(q,J＝5.6Hz ,4H),1.38(q,J＝5.9Hz,2H).HRMS(m/z):471.2119[M+H] ⁺ .

Embodiment 21

¹ H NMRδ8.07(d,J＝2.6Hz,1H),7.74(dd,J＝9.0,2.6Hz,1H),7.46(td,J＝8.0,6.0Hz,1H),7.36-7.29(m, 3H),7.24(d,J＝9.0Hz,1H),7.18(td,J＝8.8,2.6Hz,1H),6.87(td,J＝16.2,10.3Hz,1H),6.21(dd,J＝16.9 ,5.9Hz,1H ),5.78-5.75(m,1H),5.24(s,2H),4.35(dd,J＝45.6,13.4Hz,2H),3.99(d,J＝13.8Hz,1H),3.95(s,3H) ,3.46(d,J＝14.0Hz,2H),3.28-2.86(m,2H),2.20(s,3H),2.17-2.55(m,4H),1.25(s,3H).MS:(m/ z):589.21[M+H] ⁺ .

Example 22

¹ H NMRδ8.08(d,J＝2.6Hz,1H),7.75(dd,J＝9.0,2.6Hz,1H),7.46(td,J＝8.0,6.0Hz,1H),7.37-7.28(m, 3H),7.25(d,J＝9.0Hz,1H),7.17(td,J＝8.8,2.6Hz,1H),6.86(td,J＝16.2,10.3Hz,1H),6.22(dd,J＝16.9 ,5.9Hz,1H ),5.78-5.74(m,1H),5.24(s,2H),4.35(dd,J＝45.6,13.4Hz,2H),3.99(d,J＝13.8Hz,1H),3.97(s,3H) ,3.46(d,J＝14.0Hz,2H),3.29-2.86(m,2H),2.21(s,3H),2.17-2.54(m,4H),1.24(s,3H).MS:(m/ z):588.22[M+H] ⁺ .

Example 23 Biological Activity Assay

Determination of the inhibitory effect of the compounds of the present invention on EGFR (L858R + T790M) mutant kinase:

Operation process:

(1) Prepare 1× Kinase buffer.

(2) Preparation of compound concentration gradient: Dilute compound to 100 times the final maximum inhibitor concentration required in the reaction, dilute in 100% DMSO. Transfer 100 μl of this compound dilution to a well in a 96-well plate. Add 100 μl of 100% DMSO to two empty wells to perform a no-compound control and a no-enzyme control in the same 96-well plate. Label the plate as the source plate. Transfer 40 μL of compound from the source plate to a new 384-well echo plate as an intermediate plate. Transfer 100 nL of each well from the 384-well echo plate to the 384-well assay plate by echo.

(3) Add 5 μL of kinase solution to each well of the assay plate, except for the control wells without enzyme (add 5 μL of 1× kinase buffer).

(4) Prepare a substrate solution of substrate and ATP in 1x kinase reaction buffer at a concentration of 2 times the final concentration of each reagent required in the assay.

(5) Add 5 μL of substrate solution to each well of the assay plate.

(6) The 384-well assay plate was incubated at room temperature for 30 minutes or 60 minutes.

(7) Prepare a detection solution of kinase quenching buffer and antibody in Lance detection buffer at twice the desired final concentration of each reagent.

(8) Add 10 μL of the detection solution and incubate at room temperature for 60 minutes.

Table 1 lists the data (500 nM) of the activity assay of the examples of the present invention and the reference compounds. The results are shown in Table 1.

Table 1 EGFR (L858R + T790M) mutant kinase inhibition rate determination data

Test substance Inhibition rate of test substance (@500nM) BDTX-189 93.1%±1.1% Example 1 94.4%±0.4% Example 2 94.3%±0.4% Embodiment 11 94.7%±2.1% Example 12 81.9%±2.7% Embodiment 14 80.4%±1.1% Embodiment 15 84.8%±0.8% Embodiment 17 96.7%±1.1% Embodiment 18 84.7%±0.5% Embodiment 19 97.1%±1.6% Embodiment 20 87.1%±1.6% Embodiment 21 98.2%±1.1% Example 22 98.9%±1.2%

Determination of the anti-proliferative effect of the compound of the present invention combined with cetuximab on human lung adenocarcinoma cells (NCI-H1975 cells):

Operation process:

(1) Prepare 1640 complete medium in the following proportions: 1640 basal medium 89%, fetal bovine serum 10%, and double antibody 1%.

(2) The test compound was prepared into a 5 mM stock solution using cell-grade DMSO and stored in a 4°C refrigerator until use.

(3) The frozen NCI-H1975 cells were quickly thawed in a 37°C constant temperature water bath, transferred to a centrifuge tube containing 5 mL of complete culture medium, centrifuged at 1000 r for 5 min, suspended into a single cell suspension and cultured in a T25 culture flask. When the cell density reached 80%, the cells were subcultured as follows: rinsed with PBS buffer, digested with 2 mL of trypsin for 3 min, added with 6 mL of complete culture medium to stop digestion and transferred to a 15 mL centrifuge tube, centrifuged at 1000 r for 5 min, discarded the supernatant, added 5 mL of complete culture medium, blown evenly, and transferred to a T25 culture flask for continued culture.

(4) Take the cells in the logarithmic growth phase and make a suspension, count them, dilute them to 8000 cells/well and plate them in a 96-well plate, 100 μL per well, and culture them in a 37°C constant temperature incubator for 24 h. After the cells adhere to the wall, the remaining medium in each well is aspirated, and the test compound and cetuximab (1 μg/mL) diluted in the medium to different concentrations (9 concentrations) are added. At the same time, a blank control group (complete medium only) and a positive control group (cells and medium, no drug) are set up and incubated in the incubator for 72 h.

(5) Using CCK-8 reagent, add 10 μL of CCK-8 reagent to each well, incubate in an incubator for 2-4 h, shake the plate in an ELISA reader (4 min), and measure the fluorescence (OD) value of each well at 450 nm.

(6) Graphpad Prism 5 software was used to calculate the _IC50 value according to the following formula:

The data of the cell antiproliferative activity of the examples of the present invention and the reference compounds are listed in Table 2. The results are shown in Table 2.

Table 2 Results of the cell antiproliferative activity assay of the examples and reference compounds

Test substance Test substance + cetuximab NCI-H1975 IC ₅₀ (nM) BDTX-189 13.2 Example 1 7.6 Example 2 9.1 Embodiment 11 8.2 Embodiment 17 7.1 Embodiment 18 10.7 Embodiment 19 8.6 Embodiment 21 4.4 Example 22 3.5

The results showed that all the compounds of the present invention are highly effective allosteric inhibitors of EGFR.

Embodiment 24:

Evaluation of compound stability using human liver microsomes:

The liver microsomal enzyme stability of the example compounds was compared with that of BDTX189.

Assay system: The metabolic stability of the compounds of the invention was tested using liver microsomes from male and female pools with 1 mM NADPH. The samples were analyzed using a mass spectrometer. HRMS was used to determine the peak area response ratio (corresponding to the peak area of the test compound or control divided by the peak area of the analytical internal standard) without running a standard curve. In order to detect all possible metabolites, HRMS scans were performed over 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 the addition of cofactors and samples were taken at 0, 2, 4, 8, 16, 24, 36, 48 hours. A positive control (5 μM testosterone) was incubated in parallel and samples were taken at 0, 2, 4, 8, 16, 24, 36, 48 hours.

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

Analytical methods:

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: 300μl/min autosampler: injection volume 10μl. Gradient program see Table 3.

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, as described in the present invention, Examples 1, 2, 11, 12, 14 showed a metabolic half-life of more than 20 hours, and Examples 15, 17, 18, 19, 20, 21 and 22 showed a metabolic half-life of between 13 and 20 hours, which was significantly greater than the 12-hour metabolic half-life of BDTX189. The relatively long metabolic half-life enables the present invention to have the potential to reduce medical doses and expand dosing intervals.

For compounds of the general formula (I), the linking groups and substituent groups have an important influence on the pharmacodynamic properties of the compounds. Although the present invention is illustrated by the specific examples above, it should not be construed as being limited thereto; rather, the present invention encompasses the general aspects previously disclosed. Various modifications and embodiments may be made without departing from the spirit and scope of the present invention.

The embodiments provided above are not intended to limit the scope of the present invention, and the steps described are not intended to limit the execution order thereof. Those skilled in the art may make obvious improvements to the present invention in combination with existing common knowledge, which also fall within the scope of protection defined by the claims of the present invention.

Claims (10)

1. A compound of structure shown in general formula (I) or a pharmaceutically acceptable salt thereof: Wherein, ring A is selected from benzene ring, pyridine ring, pyrazine ring, R ₁ is H, C1-4 alkyl, halogen, R ₂ is halogen; or ring A is selected from benzene ring, pyrazine ring, R ₁ is H , C1-4 alkyl, halogen, R ₂ is H; _R3 is selected from substituted or unsubstituted C6-10 aryl and C5-12 heteroaryl, and the substituents are arbitrarily selected from 1, 2 or 3 of the following substituents: H, halogen; W is -R ₄ (CH ₂ ) _m -, -O(CH ₂ ) _m -, wherein m is selected from 0, 1, 2, R ₄ is selected from alkenyl, alkynyl; Y ring is selected from C3-6 ring Alkyl, C3-12 heterocyclyl; or -WY is -OR ₅ , R ₅ is selected from C1-4 alkyl; Z is selected from: H, -NR ₆ , R ₆ is selected from H, C1-4 alkyl. 2. The compound of structure shown in general formula (I) according to claim 1 or its pharmaceutically acceptable salt, it is characterized in that, C3-12 heterocyclyl is selected from morpholine ring, piperidine ring, R ₇ is selected from C1-4 alkyl. 3. the compound of structure shown in general formula (I) according to claim 1 or its pharmaceutically acceptable salt, it is characterized in that, R ₃ is selected from: 4. the compound of structure shown in general formula (I) according to claim 1 or its pharmaceutically acceptable salt, it is characterized in that, described pharmaceutically acceptable salt is inorganic salt or organic salt; Inorganic salt comprises Hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, acid phosphate; said organic salt is selected from acetate, trifluoroacetate, propionic acid Salt, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, methanesulfonate , with sulfonate, besylate, salicylate. 5. A pharmaceutical composition, characterized in that it comprises a compound represented by general formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or diluent. 6. Use of the compound of the structure represented by general formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1-4 in the preparation of medicines for treating or preventing diseases based on EGFR. 7. Use of the compound of the structure represented by the general formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1-4 in the preparation of antitumor drugs. 8. The application according to claim 7, characterized in that, during the application, the compound of the structure represented by general formula (I) or a pharmaceutically acceptable salt thereof is used as an EGFR inhibitor. 9. The use according to claim 7, wherein the tumor is selected from breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal tract cancer, small cell lung cancer or non-small cell lung cancer. 10. The application according to claim 7, characterized in that, the compound of the structure shown in general formula (I) or a pharmaceutically acceptable salt thereof is used in combination with any of the following antineoplastic agents: (i) Antineoplastic drugs acting on DNA structure; (ii) Antineoplastic drugs that affect nucleic acid synthesis; (iii) Antineoplastic drugs that affect nucleic acid transcription; (iv) Antitumor drugs for tubulin synthesis; (v) Cell signaling pathway inhibitors such as epidermal growth factor receptor inhibitors; (vi) Anti-tumor monoclonal antibody.