Diaminopyrimidines and uses thereof

Abstract

The present application provides a kind of diaminopyrimidine compound shown in formula I, and the compound can be used as three mutant (sensitive mutation / T790M / C797S) EGFR kinase inhibitor for preventing and / or treating tumor.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and particularly relates to a kind of diaminopyrimidine compound and its pharmaceutical composition and application. BACKGROUND

[0002] Epidermal Growth Factor Receptor (EGFR), a kind of receptor protein tyrosine kinases (RPTKs), is over-activated or continuously activated in a variety of tumor cells, such as lung cancer, breast cancer, prostate cancer, etc. Most of the patients with non-small cell lung cancer (NSCLC) have EGFR overexpression, and the inhibition of EGFR can significantly improve the survival of some patients.

[0003] Common mutations of EGFR can be divided into two categories: one is drug-sensitive mutation, that is, after mutation, anti-tumor targeted drugs can be used, such as 19 exon deletion and 21 exon L858R mutation; the other is drug-resistant mutation, that is, after mutation, some anti-tumor targeted drugs are resistant, such as 20 exon T790M mutation and 20 exon C797S mutation.

[0004] The first generation of EGFR small molecule inhibitors has achieved significant clinical efficacy in patients carrying EGFR sensitive mutations, and has prolonged survival. However, after using the drug for 10-12 months, most patients will develop drug resistance. Among them, more than 50% of drug-resistant patients are due to T790M secondary mutation of EGFR. Compared with L858R sensitive mutation of EGFR, L858R / T790M secondary mutation of EGFR has stronger affinity for ATP, and the first generation of drugs are all ATP competitive inhibitors, thus leading to drug resistance.

[0005] The second generation of EGFR irreversible inhibitors, although good results are obtained in preclinical studies, lack selectivity for wild-type EGFR (EGFR WT ), and have greater toxicity. Although the EGFR irreversible inhibitor Gilotrif approved by FDA in 2013 is effective for patients with advanced NSCLC carrying activating EGFR mutations (L858R, del E746-A750), at the maximum tolerated dose (MTD) in clinic, it still cannot solve the clinical drug resistance caused by EGFR T790M mutation.

[0006] The third generation of EGFR irreversible inhibitors overcomes the problem of drug resistance caused by EGFR T790MThe drug-resistant irreversible inhibitor Osimertinib (AZD9291) is clinically effective in treating patients with advanced non-small cell lung cancer with epidermal growth factor receptor T790M mutation or resistance to other EGFR inhibitors. Although Osimertinib is clinically effective in treating EGFR T790M Mutant non-small cell lung cancer has achieved great success, but some patients benefit from treatment for 9-14 months and then develop drug resistance (Nature Medicine 2015, 21(6), 560-562). Studies have found that up to 40% of drug-resistant patients develop Osimertinib resistance due to (EGFR) C797S point mutation. Further mechanism studies have shown that the (EGFR) C797S point mutation converts the cysteine at position 797 to serine, preventing Osimertinib from forming a covalent bond with the target protein, ultimately causing drug resistance. There is currently a lack of effective EGFR inhibitors for the new mutation (C797S) alone in clinical use.

[0007] Therefore, there is an urgent need for new types of highly selective EGFR inhibitors to address drug resistance and other issues caused by (EGFR) C797S point mutations. SUMMARY

[0008] The purpose of the present application is to provide a new diaminopyrimidine compound that can selectively inhibit the activity of mutant EGFR and overcome the drug resistance problem of existing EGFR tyrosine kinase inhibitors (TKI).

[0009] In a first aspect of the present application, a compound of formula I or a pharmaceutically acceptable salt thereof is provided,

[0010]

[0011] wherein,

[0012] R 1 is selected from

[0013] X a , X b , X c , X d , X e , X f , X g is independently selected from a bond, -CH2-, -CH2-CH2-, -CH2-CH2-CH2- or -CH2-CH(CH3)-;

[0014] R 1a R 1d Independently selected from C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C3-C6 cycloalkyl, and halogen-substituted C3-C6 cycloalkyl; R 1b R 1c R 1e R 1f R 1g R 1h R 1i R 1j It is independently selected from hydrogen, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C3-C6 cycloalkyl, and halogen-substituted C3-C6 cycloalkyl;

[0015] R 2a R 2b R 2c It is independently selected from hydrogen, halogen, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C1-C4 alkoxy, halogen-substituted C1-C4 alkoxy, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, C2-C4 alkenyl, halogen-substituted C2-C4 alkenyl, C2-C4 alkynyl, halogen-substituted C2-C4 alkynyl, nitro, cyano, phenyl, pyridyl;

[0016] R 3 Selected from hydrogen, halogen, methyl, halogen-substituted methyl, ethyl, halogen-substituted ethyl, methoxy, and halogen-substituted methoxy;

[0017] R 4a R 4b It is independently selected from hydrogen, halogen, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C1-C4 alkoxy, halogen-substituted C1-C4 alkoxy, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, C2-C4 alkenyl, halogen-substituted C2-C4 alkenyl, C2-C4 alkynyl, halogen-substituted C2-C4 alkynyl, nitro, cyano, phenyl, pyridyl;

[0018] The Cy group is selected from

[0019] Y a Y b Y c Y d Y e Y f Y g Y h Y i Y jIndependently selected from chemical bonds, -CH2-, -CH2-CH2-, -CH2-CH2-CH2-, or -CH2-CH(CH3)-;

[0020] R 5a R 5b R 5c R 5d R 5e R 5f R 5g R 5h Independently selected from hydrogen, halogen, methyl, halogen-substituted methyl, ethyl, halogen-substituted ethyl, methoxy, halogen-substituted methoxy;

[0021] R 7a R 7b R 7c R 7d Independently selected from hydrogen, halogen, methyl, halogen-substituted methyl, ethyl, halogen-substituted ethyl, methoxy, halogen-substituted methoxy;

[0022] R 6a R 6b R 6c R 6d R 6e R 6f R 6g R 6h R 6i R 6j It is independently selected from hydrogen, halogen, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C1-C4 alkoxy, halogen-substituted C1-C4 alkoxy, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, C2-C4 alkenyl, halogen-substituted C2-C4 alkenyl, C2-C4 alkynyl, halogen-substituted C2-C4 alkynyl, nitro, cyano, phenyl, pyridyl;

[0023] R 8a R 8b R 8c R 8d R 8e R 8f R 8i R 8j It is independently selected from hydrogen, halogen, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C1-C4 alkoxy, halogen-substituted C1-C4 alkoxy, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, C2-C4 alkenyl, halogen-substituted C2-C4 alkenyl, C2-C4 alkynyl, halogen-substituted C2-C4 alkynyl, nitro, cyano, phenyl, pyridyl;

[0024] R 9a R9c R 9e R 9g R 9i The ring is independently selected from 5-7 quintile monocyclic rings (A), 7-11 quintile spirocyclic rings (A), and 7-10 quintile bridged rings (A), wherein the monocyclic ring (A), spirocyclic ring (A), and bridged ring (A) contain one, two, or three nitrogen heteroatoms, and wherein the monocyclic ring (A), spirocyclic ring (A), and bridged ring (A) optionally contain one or two heteroatoms independently selected from oxygen and sulfur;

[0025] The C or N on the monocyclic (A), spirocyclic (A), and bridged ring (A) may optionally be independently replaced by one or more C1-C4 alkyl groups, halogen-substituted C1-C4 alkyl groups, or -(CH2) groups. m -NR 12 R 13 Substitution of 3- to 6-membered monocyclic (B) substituents;

[0026] The monocyclic ring (B) contains 0, 1, or 2 nitrogen heteroatoms; the monocyclic ring (B) contains 0, 1, or 2 heteroatoms independently selected from oxygen and sulfur; the C or N on the monocyclic ring (B) is optionally independently replaced by one or more C1-C4 alkyl groups, halogen-substituted C1-C4 alkyl groups, or -(CH2). n -NR 14 R 15 Substituents of the substituents;

[0027] m is 0, 1, or 2; R 12 R 13 It is independently selected from hydrogen, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C3-C6 cycloalkyl, and halogen-substituted C3-C6 cycloalkyl;

[0028] n is 0, 1, or 2; R 14 R 15 It is independently selected from hydrogen, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C3-C6 cycloalkyl, and halogen-substituted C3-C6 cycloalkyl;

[0029] R 10b R 10d R 10f R 10h R 10j R 11b R 11d R 11f R 11h R 11j Independently selected from hydrogen, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, -(CH2). p -NR 16 R17 ;

[0030] p is 1, 2, or 3; R 16 R 17 It is independently selected from hydrogen, C1-C4 alkyl, halogen-substituted C1-C4 alkyl, C3-C6 cycloalkyl, and halogen-substituted C3-C6 cycloalkyl.

[0031] In a preferred embodiment, R 7a R 7b R 7c R 7d It is hydrogen independently.

[0032] In a preferred embodiment, R 8a R 8b R 8c R 8d R 8e R 8f R 8i R 8j It is hydrogen independently.

[0033] In a preferred embodiment, the 5-7 elemental monocyclic ring (A) is selected from...

[0034] In a preferred embodiment, the 7-11 element helical ring (A) is selected from...

[0035] In a preferred embodiment, the 7- to 10-element bridge ring (A) is selected from...

[0036] In a preferred embodiment, the 3- to 6-membered monocyclic (B) is selected from...

[0037] In a preferred embodiment, R 1 Selected from

[0038] In a preferred example, X a X b X f X gIt is independently selected from chemical bonds, -CH2-, or -CH2-CH2-.

[0039] In a preferred embodiment, R 1a Selected from methyl, ethyl, n-propyl, isopropyl, trifluoromethyl, cyclopropyl; R 1b R 1c R 1h R 1i R 1j It is independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, trifluoromethyl, and cyclopropyl.

[0040] In a preferred embodiment, R 2a R 2b R 2c It is independently selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, trifluoromethyl, methoxy, trifluoromethoxy, and cyclopropyl.

[0041] In a preferred embodiment, R 3 Selected from hydrogen, fluorine, chlorine, bromine, methyl, trifluoromethyl, ethyl, methoxy, and trifluoromethoxy.

[0042] In a preferred embodiment, the compound

[0043] Selected from:

[0044]

[0045]

[0046]

[0047]

[0048] A second aspect of the present invention provides a pharmaceutical composition comprising:

[0049] A therapeutically effective amount of the compound as described in the first aspect of the invention or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

[0050] A third aspect of the invention provides the use of the compound or a pharmaceutically acceptable salt thereof as described in the first aspect of the invention: in the preparation of a medicament for the prevention and / or treatment of tumors. That is, for the prevention and / or treatment of tumors.

[0051] In a preferred embodiment, the tumor is a malignant tumor with an EGFR gene mutation.

[0052] In a preferred embodiment, the tumor is EGFR 敏感突变 / T790M / C797S Mutant non-small cell lung cancer; more specifically EGFR L858R / T790M / C797S Mutant non-small cell lung cancer, EGFR19del / T790M / C797S Non-small cell lung cancer.

[0053] The present invention provides diaminopyrimidine compounds or pharmaceutically acceptable salts thereof, which have the following advantages and beneficial effects:

[0054] The compounds of this invention can selectively inhibit the activity of mutant EGFR, representing a novel class of protein kinase inhibitors capable of overcoming resistance to existing EGFR tyrosine kinase inhibitors and possessing selectivity and favorable pharmacokinetic properties. These compounds can effectively inhibit the growth of various tumor cells. They can be used to prepare antitumor drugs and overcome resistance induced by existing drugs (such as gefitinib, erlotinib, and especially osimertinib), primarily targeting resistance induced by the 797 cysteine ​​mutation to serine (C797S) induced by existing third-generation EGFR small molecule inhibitors for non-small cell lung cancer, such as osimertinib (AZD9291), olmutinib (HM6171), and rociletinib (CO-1686). The compounds can be used to prevent postoperative recurrence of various tumors and for further consolidation therapy, aiming to prolong the survival of cancer patients, improve their quality of life, and inhibit tumor progression.

[0055] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0056] This invention includes the free form of compounds of formula I, as well as their pharmaceutically acceptable salts. The free form refers to the non-salt form of the compound. A pharmaceutically acceptable salt of the compounds of this invention means a conventional, non-toxic salt of the compound formed by the reaction of the compound with an inorganic / organic acid (or an inorganic / organic base). Salts of the compounds of this invention containing a basic or acidic moiety can be synthesized by conventional chemical methods; for example, salts of basic compounds can be prepared by reacting a free base with a stoichiometric amount or excess of the desired salt form of an inorganic or organic acid in a suitable solvent (or a suitable combination of solvents); similarly, for example, salts of acidic compounds can be formed by reacting with a suitable inorganic or organic base. Berg et al., Pharmaceutical Salts, Journal of Pharmaceutical Sciences, 1977:66:1-19, describe in more detail the pharmaceutically acceptable salts described above and the preparation of typical pharmaceutically acceptable salts.

[0057] Some compounds of the present invention may have asymmetric carbon atoms; racemates, single isomers, and combinations thereof are all included within the scope of the present invention.

[0058] Some compounds of the present invention may have double bonds, and both the Z configuration and the E configuration are included within the scope of the present invention.

[0059] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as deuterium. 2 H), tritium ( 3 H), Iodine-125 125 I) or C-14 14 C). All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0060] The compounds of this invention can exist in either a non-solventized or a solvated form, including hydrated forms. Generally, solvated and non-solventized forms are equivalent and both are included within the scope of this invention.

[0061] "Pharmaceutically acceptable carrier" refers to any carrier medium capable of delivering an effective amount of the active substance of the present invention without interfering with the biological activity of the active substance and without toxic side effects on the host or patient.

[0062] "Active substance," "active ingredient," or "active agent" refers to a chemical entity that can effectively treat a target disorder, disease, or symptom.

[0063] For the purposes of this study, the term "effective amount" or "therapeutic effective amount" refers to a sufficient quantity of drug or pharmacologically active agent to achieve the desired effect. The determination of the effective amount varies from person to person, depending on age and general condition, as well as the specific active substance. The appropriate effective amount in a given case can be determined by a person skilled in the art based on routine testing.

[0064] "Alkyl" refers to a branched or straight-chain saturated aliphatic hydrocarbon group with a specific number of carbon atoms. Compounds containing only carbon and hydrogen are called hydrocarbons, or simply hydrocarbons.

[0065] “C x -C y "A group refers to a group that includes x to y carbon atoms. For example, "C1-C4 alkyl" includes alkyl groups with 1, 2, 3 or 4 carbon atoms arranged in a straight chain or branched chain; including methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl and isobutyl.

[0066] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0067] "Cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group with a specific number of carbon atoms. "C3-C6 alkyl" includes cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, etc.

[0068] "Alkoxy" refers to a group with an -O-alkyl structure; for example, "C1-C4 alkoxy" refers to -O-C1-C4 alkyl.

[0069] "Alkenyl" refers to a hydrocarbon group having one or more carbon-carbon double bonds at any position in the chain. "C2-C4 alkenyl" includes, but is not limited to, vinyl, propenyl, butenyl, and butadienyl.

[0070] "Alynyl" refers to a hydrocarbon group that has one or more carbon-carbon triple bonds at any point in the chain. "C2-C4 alkynyl" includes, but is not limited to, ethynyl, propynyl, butynyl, etc.

[0071] A "ring" can be saturated, partially unsaturated, or aromatic; including but not limited to cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl, or heteroaryl; including but not limited to monocyclic, spirocyclic, or bridged rings. The number of atoms on a ring is usually defined as the ring's elemental number; for example, a "5- to 7-membered ring" refers to a ring with 5 to 7 atoms arranged in a ring.

[0072] "Spiral ring" refers to a polycyclic ring in which single rings share a single ring atom (called a spiro atom); "bridged ring" refers to a polycyclic ring in which two or more ring atoms are shared.

[0073] A ring composed of a carbon atom and at least one other atom is called a heterocycle. The atoms other than the carbon atom in the ring are called heteroatoms. Common heteroatoms are nitrogen, oxygen, and sulfur.

[0074] "Optional" or "optionally" means that the event or condition described below may occur but is not required to occur, and the description includes both the scenario in which said event or condition occurs and the scenario in which said event or condition does not occur.

[0075] The term "(substituent) substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, as long as the valence state of the particular atom is normal and the substituted compound is stable.

[0076] When any variable (e.g., R) 1 When a substance (e.g., t) appears more than once in the composition or structure of a compound, the definition of each occurrence is independent of the definitions of other occurrences. Similarly, combinations of substituents and variables are permitted, as long as such combinations stabilize the compound.

[0077] It will be understood that those skilled in the art can select the substituents and substitution patterns of the compounds of the present invention to provide chemically stable compounds that can be synthesized from readily available starting materials using techniques in the art and the methods described below. If a substituent is itself replaced by more than one group, it should be understood that these groups can be on the same carbon atom or on different carbon atoms, as long as the structure is stable.

[0078] In this invention, when referring to a specific enumerated value, the term "about" means that the value can vary from the enumerated value by no more than 1%. For example, as used herein, the expression "about 100" includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0079] The terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed. In other words, the terms also include “consistently made of” or “made of”.

[0080] The compounds of the present invention can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention.

[0081] The compounds of the present invention can be prepared by referring to the following reaction procedure:

[0082]

[0083] "Lg" stands for "leaving group," which refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (such as nucleophilic substitution). For example, representative leaving groups include halogens (such as chlorine, bromine, and iodine), sulfonate groups (such as methanesulfonate, toluenesulfonate, p-bromobenzenesulfonate, p-toluenesulfonate, etc.), and acyloxy groups (such as acetoxy, trifluoroacetoxy, etc.).

[0084] When preparing the compounds of this invention using synthetic methods well known to those skilled in the art, protecting groups may be used as needed. The term "protecting group" includes, but is not limited to, "amino protecting group," "hydroxyl protecting group," or "thiol protecting group." (Peter GMWuts and Theodora W. Greene, Greene's Protective Groups in Organic Synthesis (4)) th The book, published in 2007, systematically introduces protecting groups in organic synthesis.

[0085] Preparation Example 1

[0086] Weigh 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine (1 mmol), 2-isopropoxy-5-methyl-4-(piperidin-4-yl)-aniline (1.2 mmol), and p-toluenesulfonic acid monohydrate (1.5 mmol) into a 25 ml flask, add isopropanol (10 ml), heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0087]

[0088] 1 H NMR(400MHz,DMSO-d6)δ8.45(d,J=8.4Hz,1H),8.22(s,1H),8.03(s,1H),7.83(dd,J=7.9,1.6Hz ,1H),7.60(ddd,J=8.7,7.3,1.7Hz,1H),7.51(s,1H),7.39–7.28(m,1H),6.82(s,1H),4.54(p,J =6.0Hz,1H),3.49(p,J=6.7Hz,1H),3.09–2.95(m,2H),2.71(tt,J=11.7,3.6Hz,1H),2.61(td,J =11.9,2.6Hz,2H),2.12(s,3H),1.64–1.45(m,4H),1.22(d,J=6.0Hz,6H),1.16(d,J=6.8Hz,6H).

[0089] MS m / z(ESI,[M+H]+):558.2.

[0090] Example 1

[0091]

[0092] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-methyl-4-(4-methylpiperazin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0093] Example 2

[0094]

[0095] Weigh 100 mmol of 2-fluoro-5-nitrotoluene into a 250 mL flask, add 100 mL of dimethyl sulfoxide and stir to dissolve. Then add 150 mmol of potassium carbonate, 150 mmol of N,N,N'-trimethylethylenediamine, and 5 mL of tetrabutylammonium fluoride (1.0 M tetrahydrofuran solution). Heat the mixture to 100 °C for 6-12 h, monitoring the reaction by TLC until the reactants are completely reacted. After cooling to room temperature, pour the reaction mixture into 300 mL of water and extract three times with 400 mL of ethyl acetate. Wash the organic phase with water and saturated brine, concentrate the organic phase, and purify the concentrate by column chromatography (methanol / dichloromethane = 1 / 100 to 1 / 30) to obtain N... 1 N 1 N 2 -trimethyl-N 2 -(2-methyl-4-nitrophenyl)ethane-1,2-diamine.

[0096] Weigh N 1 N 1 N 2 -trimethyl-N 2 93 mmol of 1,2-diamine (2-methyl-4-nitrophenyl)ethane was dissolved in 180 ml methanol in a 500 ml flask by stirring at room temperature. Pd / C (20% wt) was added to displace hydrogen gas, and the mixture was stirred at room temperature for 12-24 h. The reaction was monitored by TLC until the reactants were completely reacted. Pd / C was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain N22-diamine. 1 -(2-(dimethylamino)ethyl)-N 1 ,2-Dimethylphenyl-1,4-diamine.

[0097]

[0098] Weigh out 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine and N... 1 -(2-(dimethylamino)ethyl)-N 1 2-Dimethylphenyl-1,4-diamine (0.6 mmol) and p-toluenesulfonic acid monohydrate (0.75 mmol) were placed in a 25 ml flask, and isopropanol (5 ml) was added. The mixture was heated and stirred at 85 °C for 12-18 h. The reaction was monitored by TLC until the reactants were completely reacted. After cooling to room temperature, the reaction solution was directly purified by liquid chromatography to obtain the target compound.

[0099] Example 3

[0100]

[0101] 10 mmol of 2-bromo-5-nitrotoluene, 20 mmol of N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester, and 30 mmol of sodium carbonate were weighed into a 100 mL flask. 45 mL of 1,4-dioxane and 5 mL of water were added. The mixture was stirred and purged with nitrogen. 1 mmol of bis(triphenylphosphine)-palladium dichloride was added and purged with nitrogen. The mixture was heated to 100 °C for 5-12 h. The reaction was monitored by TLC until the reactants were completely reacted. After cooling to room temperature, 100 mL of water was added to the reaction mixture. The mixture was extracted three times with 150 mL of ethyl acetate. The organic phase was washed once with saturated brine. The organic phase was concentrated and purified by column chromatography (ethyl acetate / petroleum ether = 1 / 9 to 1 / 4) to obtain t-butyl-4-(2-methyl-4-nitrophenyl)-5,6-dihydropyridine-1(2H)-carboxylic acid ester.

[0102] Weigh 9.15 mmol of t-butyl 4-(2-methyl-4-nitrophenyl)-5,6-dihydropyridine-1(2H)-carboxylic acid ester into a 100 mL flask, add 18 mL of methanol, stir to dissolve at room temperature, add Pd / C (20% wt) to replace hydrogen gas, stir to react at room temperature for 12-24 h, monitor the reaction by TLC until the starting material reacts completely, filter to remove Pd / C, concentrate the filtrate under reduced pressure to obtain t-butyl 4-(4-amino-2-methylphenyl)piperidine-1-carboxylic acid ester.

[0103]

[0104] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of t-butyl-4-(4-amino-2-methylphenyl)piperidine-1-carboxylic acid ester, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants have reacted completely, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0105] Example 4

[0106]

[0107] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 1-(4-amino-2-methylphenyl)-N,N-dimethylpiperidin-4-amine, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0108] Example 5

[0109]

[0110] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-methyl-4-(9-methyl-3,9-diazaspirocyclo[5,5]undecane-3-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0111] Example 6

[0112]

[0113] 3-Methyl-4-(7-methyl-2,7-diazaspirocyclic[3,5]nonane-3-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0114]

[0115] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 3-methyl-4-(7-methyl-2,7-diazaspirocyclo[3,5]nonane-3-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0116] Example 7

[0117]

[0118] 3-Methyl-4-(5-methylhexahydropyrrole[3,4-c]pyrrole-2(1H)-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0119]

[0120] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-methyl-4-(5-methylhexahydropyrrole[3,4-c]pyrrole-2(1H)-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 mL flask. Add 5 mL of isopropanol, heat and stir at 85 °C for 12-18 h. Monitor the reaction by TLC until the reactants are completely reacted. After cooling to room temperature, the reaction solution is directly purified by liquid chromatography to obtain the target compound.

[0121] Example 8

[0122]

[0123] 3-Methyl-4-(3-methyl-1,3-diazacycloheptane-1-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0124]

[0125] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-methyl-4-(3-methyl-1,3-diazacycloheptane-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0126] Example 9

[0127]

[0128] (S)-4-(3,4-dimethylpiperazin-1-yl)-3-methylaniline can be obtained by a similar preparation method as in Example 2.

[0129]

[0130] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of (S)-4-(3,4-dimethylpiperazin-1-yl)-3-methylaniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0131] Example 10

[0132]

[0133] (R)-4-(3,4-dimethylpiperazin-1-yl)-3-methylaniline can be obtained by a similar preparation method as in Example 2.

[0134]

[0135] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of (R)-4-(3,4-dimethylpiperazin-1-yl)-3-methylaniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0136] Example 11

[0137]

[0138] 3-Methyl-4-((3S,5R)-3,4,5-trimethylpiperazin-1-yl)-3-methylaniline can be obtained by a similar preparation method as in Example 2.

[0139]

[0140] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 3-methyl-4-((3S,5R)-3,4,5-trimethylpiperazin-1-yl)-3-methylaniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask. Add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h. Monitor the reaction by TLC until the reactants are completely reacted. After cooling to room temperature, the reaction solution is directly purified by liquid chromatography to obtain the target compound.

[0141] Example 12

[0142]

[0143] 3-Methyl-4-(4-(oxecyclobutane-3-aryl)piperazin-1-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0144]

[0145] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-methyl-4-(4-(oxecyclobutane-3-aryl)piperazin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0146] Example 13

[0147]

[0148] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 4-(4-methylpiperazin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0149] Example 14

[0150]

[0151] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 2-methyl-4-(4-methylpiperazin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0152] Example 15

[0153]

[0154] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-chloro-4-(4-methylpiperazin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0155] Example 16

[0156]

[0157] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-bromo-4-(4-methylpiperazin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0158] Example 17

[0159]

[0160] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 4-(4-methylpiperazin-1-yl)-3-trifluoromethylaniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0161] Example 18

[0162]

[0163] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 3-fluoro-4-(4-methylpiperazin-1-yl)-aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0164] Example 19

[0165]

[0166] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 3-methoxy-4-(4-methylpiperazin-1-yl)-aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0167] Example 20

[0168]

[0169] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 1-(2-(dimethylamino)ethyl)-1H-pyrazole-4-amine, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0170] Example 21

[0171]

[0172] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 1-(2-(diethylamino)ethyl)-1H-pyrazole-4-amine, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0173] Example 22

[0174]

[0175] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 4-(4-ethylpiperazin-1-yl)-3-methylaniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0176] Example 23

[0177]

[0178] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of t-butyl-4-(4-amino-2-methylphenyl)piperazine-1-carboxylic acid ester, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0179] Example 24

[0180]

[0181] 3-Methyl-4-((1S,4S)-5-methyl-2,5-diazabicyclo[2.2.1]heptane-2-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0182]

[0183] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-methyl-4-((1S,4S)-5-methyl-2,5-diazabicyclo[2.2.1]heptane-2-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask. Add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h. Monitor the reaction by TLC until the reactants are completely reacted. After cooling to room temperature, the reaction solution is directly purified by liquid chromatography to obtain the target compound.

[0184] Example 25

[0185]

[0186] (R)-t-butyl4-(4-amino-2-chlorophenyl)2-methylpiperazine-1-carboxylic acid ester can be obtained by a similar preparation method as in Example 2.

[0187]

[0188] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of (R)-t-butyl-4-(4-amino-2-chlorophenyl)2-methylpiperazine-1-carboxylic acid ester, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0189] Example 26

[0190]

[0191] 3-Chloro-4-(3-methyl-1,3-diazacycloheptane-1-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0192]

[0193] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-chloro-4-(3-methyl-1,3-diazacycloheptane-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0194] Example 27

[0195]

[0196] (R)-3-chloro-4-(3,4-dimethylpiperazin-1-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0197]

[0198] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of (R)-3-chloro-4-(3,4-dimethylpiperazin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0199] Example 28

[0200]

[0201] 3-Methyl-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0202]

[0203] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-methyl-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0204] Example 29

[0205]

[0206] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)-pyrimidine, 0.6 mmol of 4-(4-isopropylpiperazin-1-yl)-3-methylaniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0207] Example 30

[0208]

[0209] 3-Methyl-4-(4-(pyrrololin-1-yl)piperidin-1-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0210]

[0211] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-methyl-4-(4-(pyrrololin-1-yl)piperidin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0212] Example 31

[0213]

[0214] 3-Methyl-4-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)aniline can be obtained by a similar preparation method as in Example 2.

[0215]

[0216] Weigh 0.5 mmol of 2,5-dichloro-4-amino-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine, 0.6 mmol of 3-methyl-4-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)aniline, and 0.75 mmol of p-toluenesulfonic acid monohydrate into a 25 ml flask, add 5 ml of isopropanol, heat and stir at 85 °C for 12-18 h, monitor by TLC until the reactants are completely reacted, cool to room temperature, and then directly prepare the target compound by liquid-phase purification.

[0217] Examples 1 to 31: Target compounds 1 The results of H NMR and MS tests are shown in the table below:

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225] Examples 32 to 56

[0226] Following a method similar to that used in Preparation Example 1, the following compounds were obtained through Examples 32 to 56.

[0227]

[0228]

[0229]

[0230]

[0231] Example 62 Kinase Activity Assay

[0232] Kinases: EGFR-WT (catalog number PV3872), EGFR-T790M / L858R (catalog number PV4879), EGFR-C797S (catalog number A33499), and EGFR-L858R / T790M / C797S (catalog number A33502) were all purchased from Thermo Fisher Scientific.

[0233] Kinase Assay Kit: Z′-LYTE TM Kinase Assay Kit – Tyrosine 2 Peptide (item number PV3191), purchased from Thermo Fisher Scientific.

[0234] Application of Z′-LYTE TMThe technology (using fluorescence detection, enzyme-coupled, based on the difference in sensitivity of phosphorylated and non-phosphorylated peptides to protein hydrolysis) employs the fluorescence resonance energy transfer (FRET) principle and uses Z′LYTE TM FRET peptide substrates were used, and a second-order reaction was employed to detect the kinase activity of the compounds. The kinase was serially diluted and added to the FRET peptide and ATP, followed by different concentrations of the compound. After reacting for 1 hour, a site-specific protease was added to recognize and cleave the non-phosphorylated FRET peptide. After reacting for another 1 hour, the absorbance at 445 nm and 520 nm was detected using a 400 nm excitation wavelength.

[0235] The emission ratio can be obtained by dividing the 445nm detection value (Coumarin Emission) by the 520nm detection value (Fluorescein Emission).

[0236]

[0237] The degree of phosphorylation (%Phosphorylation) of the sample wells can be calculated using 0% and 100% phosphorylation control wells.

[0238]

[0239] C 100％ = Average Coumarin Emission at 100% Phosphorylation

[0240] C 0％ = Average Coumarin Emission at 0% Phosphorylation

[0241] F 100％ = Average Fluorescein Emission at 100% Phosphorylation

[0242] F 0％ =Average Fluorescein Emission at 0% Phosphorylation

[0243] Calculate IC50 based on the kinase activity versus concentration curve. 50 Value; IC of EGFR-L858R / T790M / C797S 50 See Table 1. "++" indicates IC. 50 <100nm, "++" indicates 100nm≤IC 50 <1000nm, "+" indicates IC 50 ≥1000nm.

[0244] In Table 1, Activity% represents the percentage value of the phosphorylation level when the concentration of the test compound is 1 μM compared to the phosphorylation level of the control group without the compound, and no replicate wells were set. "+++" indicates Activity% ≤ 20, "++" indicates 20 < Activity% ≤ 40, and "+" indicates 40 < Activity% ≤ 60.

[0245] As can be seen from Table 1, the compounds of the present invention exhibit high kinase inhibitory activity, especially with respect to EGFR-WT, and their inhibitory activity against triple-mutant EGFR is stronger.

[0246] Cell viability experiment of Example 63

[0247] Cell lines: A431 (human epidermal carcinoma cell line), NCI-H1975 (human non-small cell lung cancer cell line), sourced from the American Type Culture Collection (ATCC). Method: CCK8 (Cell Counting Kit-8) method.

[0248] The specific steps are as follows: Inoculate a certain number of tumor cells in the logarithmic growth phase into a 96-well culture plate. After culturing for 24 h until the cells adhere, add the test compounds of the present invention at different concentrations, set three replicate wells for each concentration, and set the corresponding concentration of DMSO solvent control and cell-free control wells. After treating the cells with the drug for 72 h, add 10 μL of CCK8 reagent, continue to culture for 1 - 4 h, and use an enzyme-labeled instrument to measure the absorbance at wavelengths of 450 nm and 650 nm. The actual absorbance value A of each well = OD 450 -OD 650 .

[0249] Cell viability (%) = [(As - Ac) / (Ab - Ac)] × 100%

[0250] As: Test compound well (containing cells and test compound)

[0251] Ab: Solvent control well (containing cells, without test compound)

[0252] Ac: Cell-free control well (without cells, without test compound)

[0253] The test compound was serially diluted 3-fold to obtain 10 concentrations ranging from 10 μM to 0.508 nM. Input the cell viability data and the concentration of the test compound into GraphPad Prism 5 Demo software, and use a non-linear regression model to calculate the IC 50 value. The results are shown in Table 2. "+++" indicates IC 50 < 100 nm, "++" indicates 100 nm ≤ IC 50 < 1000 nm, "+" indicates IC50 ≥1000nm.

[0254] As shown in Table 2, the compounds of this invention can effectively inhibit the growth of A431 and NCI-H1975 tumor cells. Table 1

[0255]

[0256] Table 2

[0257]

[0258] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound of Formula I, or a pharmaceutically acceptable salt thereof, wherein R 1 selected from X a is selected from a chemical bond, -CH2-; R 1a selected from methyl, ethyl, n-propyl, i-propyl; R 2a , R 2b , R 2c is independently selected from hydrogen; R 3 selected from hydrogen; R 4b selected from hydrogen; R 4a selected from halogen; The group Cy is selected from Y a is selected from a chemical bond, -CH2-; R 5a is hydrogen; R 7a is hydrogen; R 6a selected from hydrogen, halogen, C1-C4alkyl; R 8a is hydrogen; R 9a selected from a 5-7 membered monocyclic ring (A), a 7-11 membered spiro ring (A), a 7-10 membered bridged ring (A), said monocyclic ring (A), spiro ring (A), bridged ring (A) containing 1 or 2 or 3 nitrogen heteroatoms; the C or N on the monocycle (A), spirocycle (A), bridgcycle (A) is optionally substituted with one or more substituents independently selected from C1-C4 alkyl, 3-6 membered monocycle (B); the monocycle (B) contains 0 or 1 nitrogen heteroatom; the C or N on the monocycle (B) is optionally substituted with one or more substituents independently selected from C1-C4 alkyl; The 5-7 membered monocyclic ring (A) is selected from The 7-11 membered spirocyclic ring (A) is selected from The 7-10 membered bridged ring (A) is selected from The 3-6 membered monocyclic ring (B) is selected from 2. A compound selected from the group consisting of:

3. A pharmaceutical composition, characterized by, the pharmaceutical composition comprises: a therapeutically effective amount of a compound of any one of claims 1-2, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

4. Use of a compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention and / or treatment of a tumor, characterized in that, The tumor is EGFR 敏感突变 / T790M / C797S Mutated non-small cell lung cancer.

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