CN113527335B - Macrocyclic compounds as EGFR inhibitors and their applications - Google Patents

Abstract

The present invention belongs to the field of medicinal chemistry and relates to a class of macrocyclic compounds used as EGFR inhibitors and their applications. Specifically, the present invention provides compounds represented by formula (I) or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, their preparation methods, and pharmaceutical compositions containing these compounds and the use of these compounds or compositions for treating EGFR-mediated diseases.

Description

Macrocyclic compound as EGFR inhibitor and application thereof

Technical Field

The invention belongs to the field of medicinal chemistry, and in particular relates to a macrocyclic compound serving as an EGFR inhibitor or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, a preparation method of the macrocyclic compound or the isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, a pharmaceutical composition containing the macrocyclic compound and application of the macrocyclic compound or the isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug in treating EGFR-mediated diseases.

Background

EGFR (Epidermal Growth Factor Receptor) are receptors for Epidermal Growth Factor (EGF) cell proliferation and signaling. EGFR belongs to one of the ErbB receptor families, which includes EGFR (ErbB-1), HER2/c-neu (ErbB-2), HER 3 (ErbB-3) and HER 4 (ErbB-4). EGFR, also known as HER1, erbB1, mutations or overexpression generally causes tumors. EGFR is a glycoprotein, belongs to tyrosine kinase type receptor, has a through cell membrane and has a molecular weight of 170kDa.

EGFR is related to proliferation of tumor cells, angiogenesis, tumor invasion, metastasis and inhibition of apoptosis, and research shows that EGFR high expression or abnormal expression exists in solid tumors such as glial cells, kidney cancer, lung cancer, prostate cancer, pancreatic cancer, breast cancer and the like. Currently about 30% -40% of asian NSCLC patients carry EGFR mutations at the time of diagnosis.

Common mutations of EGFR can be divided into two main classes, one is drug-sensitive mutation, i.e. anti-tumor targeted drugs such as 19 exon deletion and 21 exon L858R mutation can be used after mutation, and the other is drug-resistant mutation, i.e. drug resistance to a certain anti-tumor targeted drug such as 20 exon T790M mutation and 20 exon C797S mutation after mutation. AZD9291 (Aoditinib) is an oral small molecule third generation EGFR-TKI, which is a lung cancer drug aiming at EGFR T790M mutation, but partial benefited patients have drug resistance after 9-14 months of treatment. It was found that up to 40% of drug resistant patients were resistant to octenib due to egfr_c797S point mutation. Further mechanism studies indicate that point mutation of egfr_c797s converts cysteine at position 797 to serine, resulting in failure of the formation of covalent binding of octreotide to the target protein, C797S being an important cause of drug resistance in the third generation drug octreotide. There is currently no clinically effective EGFR inhibitor for the novel mutation (C797S) alone, and therefore there is a need to develop a new generation EGFR inhibitor for mutations involving C797S.

Disclosure of Invention

It is an object of the present invention to provide a compound having EGFR inhibitory activity represented by the general formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof,

Wherein,

Cy is selected from aryl, heteroaryl, cycloalkyl, heterocyclyl, and heterocycloheteroaryl, the aryl, heteroaryl, cycloalkyl, heterocyclyl, and heterocycloheteroaryl being optionally substituted with one or more groups selected from halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, monoalkylamino, alkylamido, alkanoyl, alkylsulfonyl, aminoacyl, alkylaminoacyl, dialkylamino, alkenyl, alkynyl, haloalkoyl, hydroxyalkylacyl, cycloalkylacyl, heterocycloalkylacyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and oxo groups;

L is selected from the group consisting of a bond, -S-, -O-, -CH ₂-、-CH₂CH₂、-C(O)-、-S(O)-、-S(O)₂ -, Wherein R ⁴、R⁵ is independently selected from hydrogen, halogen, hydroxy, carboxy, cyano, amino, alkenyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy;

R ¹ is selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkylamido, alkylacyl, aminoacyl, alkylaminoacyl, and dialkylamino, and

R ²、R³ is independently selected from the group consisting of hydrogen, halogen, hydroxy, carboxy, cyano, amino, alkenyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, monoalkylamino, alkanoylamino, alkanoyl, aminoacyl, alkylaminoacyl, and cycloalkyl.

It is a further object of the present invention to provide a process for the preparation of the compounds of formula (I) of the present invention or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof.

It is a further object of the present invention to provide compositions comprising a compound of formula (I) of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof and a pharmaceutically acceptable carrier, as well as compositions comprising a compound of formula (I) of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof and another drug or drugs.

It is a further object of the present invention to provide a method of treating EGFR mediated diseases by the compounds of formula (I) or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof, and the use of the compounds of formula (I) or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof, in the manufacture of a medicament for treating EGFR mediated diseases.

Aiming at the purpose of the invention, the invention provides the following technical scheme:

in a first aspect, the present invention provides a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof,

Wherein,

Cy is selected from aryl, heteroaryl, cycloalkyl, heterocyclyl, and heterocycloheteroaryl, the aryl, heteroaryl, cycloalkyl, heterocyclyl, and heterocycloheteroaryl being optionally substituted with one or more groups selected from halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, monoalkylamino, alkylamido, alkanoyl, alkylsulfonyl, aminoacyl, alkylaminoacyl, dialkylamino, alkenyl, alkynyl, haloalkoyl, hydroxyalkylacyl, cycloalkylacyl, heterocycloalkylacyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and oxo groups;

L is selected from the group consisting of a bond, -S-, -O-, -CH ₂-、-CH₂CH₂、-C(O)-、-S(O)-、-S(O)₂ -, Wherein R ⁴、R⁵ is independently selected from hydrogen, halogen, hydroxy, carboxy, cyano, amino, alkenyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy;

R ¹ is selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxy, cyano, amino, monoalkylamino, alkylamido, alkylacyl, aminoacyl, alkylaminoacyl, and dialkylamino, and

R ²、R³ is independently selected from the group consisting of hydrogen, halogen, hydroxy, carboxy, cyano, amino, alkenyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, monoalkylamino, alkanoylamino, alkanoyl, aminoacyl, alkylaminoacyl, and cycloalkyl.

In some preferred embodiments, the compounds of the invention are compounds of formula (I) or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof, wherein:

Cy is selected from the group consisting of C _6-12 aryl, 5-12 membered heteroaryl, C _3-12 cycloalkyl, 3-12 membered heterocyclyl and 5-12 membered heteroaryl, said C _6-12 aryl, 5-12 membered heteroaryl, C _3-12 cycloalkyl, 3-12 membered heterocyclyl and 5-12 membered heteroaryl are optionally substituted with one or more groups selected from halogen, hydroxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, C _1-6 alkoxy, Halogenated C _1-6 alkoxy, hydroxyC _1-6 alkoxy, nitro, carboxyl, cyano, amino, mono C _1-6 alkylamino, C _1-6 alkylamido, C _1-6 Alkylacyl, C _1-6 alkylsulfonyl, aminoacyl, C _1-6 alkylaminoacyl, di C _1-6 alkylamino, C _2-10 alkenyl, C _2-10 alkynyl, halogenated C _1-6 alkanoyl, hydroxyC _1-6 alkanoyl, C _3-12 cycloalkylacyl, 3-12 membered heterocyclylacyl, C _3-12 cycloalkyl, 3-12 membered heterocyclyl, 6-12 membered aryl, 5-12 membered heteroaryl, and oxo groups;

Further preferred Cy is selected from the group consisting of C _6-10 aryl, 5-10 membered heteroaryl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl and 5-10 membered heteroaryl, said C _6-10 aryl, 5-10 membered heteroaryl, C _3-10 cycloalkyl, 3-10 heterocyclyl and 5-10 heteroaryl are optionally substituted with one or more substituents selected from halogen, hydroxy, C _1-3 alkyl, halo C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, haloC _1-3 alkoxy, hydroxyC _1-3 alkoxy, nitro, carboxyl, cyano, amino, mono C _1-3 alkylamino, C _1-3 Alkylacylamino, C _1-3 Alkylacyl, C _1-3 alkylsulfonyl, aminoacyl, C _1-3 Alkylaminoacyl, Bis C _1-3 alkylamino, C _2-6 alkenyl, C _2-6 alkynyl, halo C _1-3 alkanoyl, hydroxy C _1-3 alkanoyl, Group substitution of C _3-8 cycloalkylacyl, 3-8 membered heterocyclylacyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, 6-8 membered aryl, 5-8 membered heteroaryl, and oxo groups;

Still more preferably, cy is selected from phenyl, 5-6 membered heteroaryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 3-10 membered azetidinyl and 4-8 membered azetidinyl and 5-8 membered heteroaryl, said Cy is selected from phenyl, 5-6 membered heteroaryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 3-10 membered heterocyclyl and 4-8 membered heterocyclyl and 5-8 membered heteroaryl optionally being substituted with one or more groups selected from halogen, hydroxy, C _1-3 alkyl, halogenated C _1-3 alkyl, Hydroxy C _1-3 alkyl, C _1-3 alkoxy, halo C _1-3 alkoxy, hydroxy C _1-3 alkoxy, Nitro, carboxyl, cyano, amino, mono C _1-3 alkylamino, C _1-3 alkylamido, C _1-3 alkanoyl, C _1-3 alkylsulfonyl, aminoacyl, C _1-3 alkylamino acyl, di C _1-3 alkylamino, C _2-6 alkenyl, C _2-6 alkynyl, Halogenated C _1-3 Alkylacyl, hydroxyC _1-3 Alkylacyl, C _3-8 Cycloalkylacyl, 3-8 membered heterocyclylacyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, 6-8 membered aryl, 5-8 membered heteroaryl, and oxo groups.

In some specific embodiments, a compound of formula (I) according to the invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein Cy is selected from phenyl, 5-6 membered heteroaryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 3-10 membered azetidinyl and 4-8 membered azetidinyl and 5-8 membered heteroaryl, optionally substituted with one or more groups selected from fluoro, chloro, bromo, hydroxy, methyl, ethyl, propyl, trifluoromethyl, hydroxymethyl, methoxy, nitro, carboxy, cyano, amino, aminomethyl, formylamino, formyl, methylsulfonyl, aminoacyl, methylaminoacyl, dimethylamino, cyclopropyl, cyclobutyl, oxetanyl, aziridinyl, oxetanyl, azetidinyl, oxo.

In some preferred embodiments, the compounds of the invention are compounds of formula (I) or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof, wherein:

R ¹ is selected from the group consisting of hydrogen, halogen, hydroxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, C _1-6 alkoxy, halo C _1-6 alkoxy, hydroxy C _1-6 alkoxy, nitro, carboxy, cyano, amino, mono C _1-6 alkylamino, C _1-6 alkylamido, C _1-6 alkanoyl, aminoacyl, C _1-6 alkylaminoacyl, and di C _1-6 alkylamino;

Further preferably, R ¹ is selected from the group consisting of hydrogen, halogen, hydroxy, C _1-3 alkyl, halo C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, halo C _1-3 alkoxy, hydroxy C _1-3 alkoxy, nitro, carboxy, cyano, amino, mono C _1-3 alkylamino, C _1-3 alkylamido, C _1-3 alkanoyl, aminoacyl, C _1-3 alkylaminoacyl, and di C _1-3 alkylamino;

still more preferably, R ¹ is selected from the group consisting of hydrogen, halogen, hydroxy, methyl, ethyl, propyl, isopropyl, halogenated C _1-3 alkyl, hydroxyC _1-3 alkyl, C _1-3 alkoxy, halogenated C _1-3 alkoxy, hydroxyC _1-3 alkoxy, nitro, carboxyl, cyano, amino, mono C _1-3 alkylamino, C _1-3 alkylamido, C _1-3 alkanoyl, aminoacyl, C _1-3 alkylaminoacyl, and di C _1-3 alkylamino.

In some preferred embodiments, the compounds of the invention are compounds of formula (I) or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof, wherein:

R ²、R³ is independently selected from the group consisting of hydrogen, halogen, hydroxy, carboxy, cyano, amino, C _1-3 alkyl, halo C _1-3 alkyl, hydroxy C _1-3 alkyl, C _1-3 alkoxy, mono C _1-3 alkylamino, C _1-3 alkylamido, C _1-3 alkanoyl, aminoacyl, C _1-3 alkylaminoacyl, and C _3-6 cycloalkyl;

Further preferably, R ²、R³ is independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, iodo, hydroxy, carboxy, cyano, amino, methyl, ethyl, propyl, halogenated C _1-3 alkyl, hydroxyc _1-3 alkyl, C _1-3 alkoxy, mono C _1-3 alkylamino, C _1-3 alkylamido, C _1-3 alkanoyl, aminoacyl, C _1-3 alkylaminoacyl, and C _3-6 cycloalkyl.

In some preferred embodiments, the present invention provides compounds of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein formula (I) has the structure of formula (II),

Wherein R ¹、R³, cy, L have the definitions described above for formula (I).

In some preferred embodiments, a compound of formula (I) or formula (II) according to the invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein L is selected from the group consisting of a bond, -CH ₂-、-CH₂CH₂、-C(O)-、-S(O)₂ -

In some embodiments, a compound of formula (I) or formula (II) according to the invention above, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein Cy is selected from Which is optionally substituted with one or more groups selected from halogen, hydroxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, C _1-6 alkoxy, halo C _1-6 alkoxy, hydroxy C _1-6 alkoxy, nitro, carboxy, cyano, amino, mono C _1-6 alkylamino, C _1-6 alkylamido, C _1-6 alkanoyl, C _1-6 alkylsulfonyl, aminoacyl, C _1-6 alkylaminoacyl, bis C _1-6 alkylamino, C _2-10 alkenyl, C _2-10 alkynyl, halo C _1-6 alkanoyl, hydroxy C _1-6 alkanoyl, C _3-12 cycloalkylacyl, 3-12 membered heterocyclylacyl, C _3-12 cycloalkyl, 3-12 membered heterocyclyl, 6-12 membered aryl, 5-12 membered heteroaryl and oxo.

In some preferred embodiments, a compound of formula (I) or formula (II) according to the invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein Cy-L is selected from

In some embodiments, the present invention provides compounds of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein formula (I) has the structure of formula (III) below,

Wherein R ¹、R²、R³, L have the definition set forth in formula (I) above, ring A is selected from 3-12 membered azacycloalkyl, 3-12 membered diazacycloalkyl, 3-12 membered azacyclyl, 3-12 membered diazacyclyl, 5-12 membered azaheteroaryl, R ⁶ is each independently selected from halogen, hydroxy, carboxy, cyano, amino, alkenyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, monoalkylamino, alkylamido, alkylacyl, aminoacyl, alkylaminoacyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, n is 1,2,3,4, 5 or 6, or two R ⁶ together with the atoms to which they are attached form 3-12 membered azacyclyl, 3-12 membered diazacyclyl, 3-12 membered azaheterocyclyl, 5-12 membered azaheteroaryl, 3-12 membered dioxacycloalkyl, 3-12 membered dioxacyclyl, 3-12 membered oxaheterocyclyl, 3-12 membered dioxaheterocyclyl, 3-12 membered dioxacyclyl or 5-membered dioxacyclyl, optionally substituted with halogen, acylamino, hydroxy, alkylaminoyl, acylamino, alkyl, acylamino, hydroxy, alkylaryl, acylamino, alkyl, alkylamino, acylamino, or an alkyl group.

In some embodiments, a compound of formula (III) according to the invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein ring A is selected from 3-8 membered azacycloalkyl, 3-8 membered diazacycloalkyl, 3-8 membered azacyclyl, 3-8 membered diazacyclyl, 5-12 membered azaheteroaryl, R ⁶ is each independently selected from halogen, hydroxy, carboxy, cyano, amino, C _2-6 alkenyl, C _1-6 alkyl, haloC _1-6 alkyl, hydroxyC _1-6 alkyl, C _1-6 alkoxy, mono C _1-6 alkylamino, C _1-6 Alkylacylamino, C _1-6 Alkylacyl, aminoacyl, C _1-6 Alkylaminoacyl, C _6-12 aryl, C _5-12 heteroaryl, C _3-8 cycloalkyl, C _3-8 heterocycloalkyl, n being 2, 3 or 4, or two R ⁶ together with the atom to which they are attached form a 3-8 membered azacycloalkyl, 3-8 membered diazacycloalkyl, 3-8 membered azacyclyl, 3-8 membered diazacyclyl, 5-12 membered azaheteroaryl, 3-8 membered oxacycloalkyl, 3-8 membered dioxacycloalkyl, 3-8 membered oxaheterocyclyl, 3-8 membered dioxacyclyl or 5-12 membered oxaheteroaryl, optionally substituted with a member selected from halogen, hydroxy, carboxy, cyano, amino, C _2-6 alkenyl, C _1-6 alkyl, halogenated C _1-6 alkyl, hydroxyC _1-6 alkyl, C _1-6 alkoxy, mono C _1-6 alkylamino, C _1-6 alkylamido, C _1-6 Alkylacyl, aminoacyl, C _1-6 Alkylaminoacyl, C _6-12 aryl, C _5-12 heteroaryl, The groups of C _3-8 cycloalkyl and C _3-8 heterocycloalkyl are substituted.

In some embodiments, a compound of formula (III) according to the invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein ring A is selected from 3-8 membered azacycloalkyl, 3-8 membered diazacycloalkyl, 3-8 membered azacyclyl, 3-8 membered diazacyclyl, 5-12 membered azaheteroaryl, R ⁶ is each independently selected from halogen, hydroxy, carboxy, cyano, amino, C _2-6 alkenyl, C _1-6 alkyl, haloC _1-6 alkyl, hydroxyC _1-6 alkyl, C _1-6 alkoxy, mono C _1-6 alkylamino, C _1-6 Alkylacylamino, C _1-6 Alkylacyl, aminoacyl, C _1-6 Alkylaminoacyl, C _6-12 aryl, C _5-12 heteroaryl, C _3-8 cycloalkyl, C _3-8 heterocycloalkyl, n being 2, 3 or 4, or two R ⁶ together with the atom to which they are attached form a 3-8 membered azacycloalkyl, 3-8 membered diazacycloalkyl, 3-8 membered azacyclyl, 3-8 membered diazacyclyl, 5-12 membered azaheteroaryl, 3-8 membered oxacycloalkyl, 3-8 membered dioxacycloalkyl, 3-8 membered oxaheterocyclyl, 3-8 membered dioxacyclyl or 5-12 membered oxaheteroaryl, optionally substituted with a member selected from the group consisting of fluoro, chloro, bromo, hydroxy, methyl, ethyl, propyl, trifluoromethyl, hydroxymethyl, methoxy, nitro, carboxy, cyano, amino, aminomethyl, formylamino, formyl, methylsulfonyl, aminoacyl, methylaminoacyl, dimethylamino, cyclopropyl, cyclobutyl, oxetanyl, aziridinyl, oxetanyl, one or more of the azetidinyl, oxo groups are substituted.

In some embodiments, a compound of formula (III) according to the invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein ring a is selected fromR ⁶ is independently selected from one or more of fluorine, chlorine, bromine, hydroxyl, methyl, ethyl, propyl, trifluoromethyl, hydroxymethyl, methoxy, nitro, carboxyl, cyano, amino, aminomethyl, formylamino, formyl, methylsulfonyl, aminoacyl, methylaminoacyl, dimethylamino, cyclopropyl, cyclobutyl, oxetanyl, aziridinyl, oxetanyl, azetidinyl, oxo groups substituted, n is 2,3 or 4; or two R ⁶ together with the atoms to which they are attached form a 3-8 membered azacycloalkyl, 3-8 membered diazacycloalkyl, 3-8 membered azacyclyl, 3-8 membered diazacyclyl, 5-12 membered azaheteroaryl, 3-8 membered oxacycloalkyl, 3-8 membered dioxacycloalkyl, 3-8 membered oxaheterocyclyl, 3-8 membered dioxacyclyl or 5-12 membered oxaheteroaryl group optionally substituted with one or more groups selected from fluoro, chloro, bromo, hydroxy, methyl, ethyl, propyl, trifluoromethyl, hydroxymethyl, methoxy, nitro, carboxy, cyano, amino, aminomethyl, formylamino, formyl, methylsulfonyl, aminoacyl, methylaminoacyl, dimethylamino, cyclopropyl, cyclobutyl, oxetanyl, aziridinyl, oxetanyl, azetidinyl, oxo groups.

The present invention provides the following specific compounds or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof:

In another aspect, the present invention provides a process for the preparation of a compound of formula (I) according to the invention comprising:

1) The compound of formula 3 may be prepared by reacting a compound of formula 1 with a compound of formula 2,

2) The compound of formula 4 may be prepared by reacting a compound of formula 3,

3) The compound of formula 6 may be prepared by reacting a compound of formula 4 with a compound of formula 5,

4) The compound of formula 7 may be prepared by reduction of a compound of formula 6,

5) The compound of formula 8 may be prepared by reacting a compound of formula 7,

6) The compound of formula 9 may be prepared by hydrolysis of a compound of formula 8,

7) The compound of formula 10 may be prepared by reacting a compound of formula 9,

8) The compounds of formula (I) may be prepared by reacting compounds of formula 10.

Wherein R ¹、R²、R³, cy, L have the definitions described in the general formula (I), and the compound of the formula (2) and the compound of the formula (5) are commercially available compounds or can be synthesized by other technical means which are customary to those skilled in the art.

In a third aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof.

In some embodiments, the invention provides a compound of the invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, and a pharmaceutical composition comprising a compound of the invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, for use in the treatment of an EGFR mediated disease.

In some embodiments, the present invention provides pharmaceutical compositions comprising a compound of the present invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, and a pharmaceutically acceptable carrier.

The compounds of the present invention, or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof, may be admixed with a pharmaceutically acceptable carrier, diluent or excipient to prepare a pharmaceutical formulation suitable for oral or parenteral administration. Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and oral routes. The formulation may be administered by any route, for example by infusion or bolus injection, by absorption through the epithelial or skin mucosa (e.g. oral mucosa or rectum, etc.). Administration may be systemic or local. Examples of formulations for oral administration include solid or liquid dosage forms, specifically including tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions and the like. The formulations may be prepared by methods known in the art and comprise carriers, diluents or excipients conventionally used in the art of pharmaceutical formulations.

In a fourth aspect, the present invention provides a method for treating an EGFR-mediated disease, and the use of a compound of formula (I), (II) or (III) of the present invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, or a pharmaceutical composition comprising the same, in the manufacture of a medicament for treating an EGFR-mediated disease.

In some preferred embodiments, the present invention provides methods for treating EGFR-mediated diseases, including but not limited to cancer, proliferative diseases, metabolic diseases or hematological diseases, using the compounds of formula (I), (II) or (III) of the invention, or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs thereof, or pharmaceutical compositions comprising the same, and in the manufacture of a medicament for treating EGFR-mediated diseases. In some embodiments, the EGFR-mediated disease described herein is cancer.

In some preferred embodiments, the invention provides methods of treating EGFR-mediated diseases including, but not limited to, breast cancer, esophageal cancer, bladder cancer, lung cancer (e.g., bronchogenic carcinoma, small Cell Lung Cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma, lung squamous carcinoma, hematopoietic system carcinoma, lymphoma, medulloblastoma, rectal adenocarcinoma, colon cancer, gastric cancer, pancreatic cancer, liver cancer, adenoid cystic carcinoma, prostate cancer, head and neck squamous cell carcinoma, brain cancer, hepatocellular carcinoma, melanoma, oligodendroglioma, glioblastoma, ovarian clear cell carcinoma, ovarian serous adenocarcinoma, thyroid cancer, multiple myeloma (AML), renal cell carcinoma, triple-negative breast cancer, lymphoma, or a pharmaceutical composition comprising the compound of formula (I), (II) or (III) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof of the invention, or a pharmaceutical composition comprising the compound of formula (I), (II) or (III).

The compound has specific tyrosine kinase inhibition activity on mutant drug-resistant EGFR, such as del19, T790M, C797S, L858R mutant drug-resistant EGFR, is used as a mutant drug-resistant EGFR specific tyrosine kinase inhibitor, and has better prevention and/or treatment effects on diseases mediated by the drug-resistant mutant EGFR, such as cancers, proliferative diseases, metabolic diseases or blood diseases. In some embodiments, the mutant drug resistant EGFR is a Del19/T790M/C797S or L858R/T790M/C797S triple mutation. In other embodiments, the mutant drug-resistant EGFR is del19/T790M, L858R/T790M, L858R or del19 primary or secondary mutation.

Definition of terms

Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

The terms "hydrogen", "carbon", "oxygen" in the compounds of the present invention include all isotopes thereof. Isotopes are understood to include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include protium, tritium, and deuterium, isotopes of carbon include ¹²C、¹³ C and ¹⁴ C, isotopes of oxygen include ¹⁶ O and ¹⁸ O, and the like.

"Isomer" in the present invention refers to molecules of the same atomic composition and manner of attachment, but differing in their three-dimensional spatial arrangement, including, but not limited to, diastereomers, enantiomers, cis-trans isomers, and mixtures thereof, such as racemic mixtures. Many organic compounds exist in optically active form, i.e. they have the ability to rotate the plane of plane polarized light. In describing optically active compounds, the prefix D, L or R, S is used to denote the absolute configuration of the chiral center of the molecule. The prefix D, L or (+), (-) is used to name the sign of the compound plane polarization rotation, where (-) or L means that the compound is left-handed and the prefix (+) or D means that the compound is right-handed. The chemical structures of these stereoisomers are identical, but the stereoisomers are not identical. The particular stereoisomer may be an enantiomer, and the mixture of isomers is commonly referred to as an enantiomeric mixture. The 50:50 enantiomeric mixture is known as a racemic mixture or racemate, which may result in the absence of stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to a mixture of two enantiomers in equimolar amounts, lacking optical activity.

Depending on the choice of starting materials and methods, the compounds of the invention may be present in the form of one of the possible isomers or mixtures thereof, for example racemates and non-corresponding isomer mixtures, depending on the number of asymmetric carbon atoms. Optically active (R) -or (S) -isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.

The resulting mixture of any stereoisomers may be separated into pure or substantially pure geometric isomers, enantiomers, diastereomers, e.g., by chromatography and/or fractional crystallization, depending on the differences in the physicochemical properties of the components.

The "halogen" in the present invention means fluorine, chlorine, bromine, iodine. "halo" in the present invention means substituted with fluorine, chlorine, bromine or iodine.

"Alkyl" in the present invention refers to a straight or branched saturated aliphatic hydrocarbon group, preferably a straight or branched group having 1 to 6 carbon atoms, further preferably a straight or branched group having 1 to 3 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl and the like. Alkyl groups may be substituted or unsubstituted, and when substituted, the substituents may be at any useful point of attachment.

The term-C (O) -as used herein means "carbonyl".

The term-S (O) ₂ -as used herein means "sulfonyl".

"Haloalkyl" in the present invention refers to an alkyl group substituted with at least one halogen.

"Hydroxyalkyl" in the present invention refers to an alkyl group substituted with at least one hydroxy group.

"Alkoxy" in the present invention refers to an-O-alkyl group. Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, isobutoxy, sec-butoxy and the like. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents may be at any useful point of attachment.

"Cycloalkyl" in the present invention refers to a cyclic saturated hydrocarbon group. Suitable cycloalkyl groups may be substituted or unsubstituted, monocyclic, bicyclic or tricyclic saturated hydrocarbon groups having 3 to 12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

"Heterocyclyl" of the present invention refers to groups of 3-to 12-membered non-aromatic ring systems ("3-12 membered heterocyclyl") having 1 to 4 ring heteroatoms, each of which is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon. In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, as long as the valence permits. The heterocyclyl groups may either be monocyclic ("monocyclic heterocyclyl") or fused, bridged or spiro ring systems (e.g., bicyclic systems (also known as "bicyclic heterocyclyl")) and may be saturated or may be partially unsaturated. Suitable heterocyclyl groups include, but are not limited to, piperidinyl, azetidinyl, aziridinyl, tetrahydropyrrolyl, piperazinyl, dihydroquinazolinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, Etc. Each instance of a heterocyclic group may be optionally substituted or unsubstituted, and when substituted, the substituent may be at any useful point of attachment.

"Aryl" in the present invention refers to aromatic systems which may comprise a single ring or a fused multiple ring, preferably a single ring or a fused double ring, containing from 6 to 12 carbon atoms, preferably from about 6 to about 10 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, fluorenyl, indanyl. Aryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents may be at any useful point of attachment.

"Heteroaryl" according to the invention means an aryl group having at least one carbon atom replaced by a heteroatom, preferably consisting of 5 to 12 atoms (5 to 12 membered heteroaryl), more preferably 5 to 10 atoms (5 to 10 membered heteroaryl), said heteroatom being O, S, N. The heteroaryl group includes, but is not limited to, imidazolyl, pyrrolyl, furanyl, thienyl, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl, indolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, isoindolyl, benzopyrazolyl, benzimidazolyl, benzofuranyl, benzopyranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, quinolinyl, isoquinolinyl, quinazolinyl, cinnolinyl, quinoxalinyl, benzoxazinyl, benzothiazinyl, imidazopyridinyl, pyrimidopyrazyl, pyrimidoimidazolyl, and the like. Heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents may be at any useful point of attachment.

The term "pharmaceutically acceptable salts" as used herein refers to salts of the compounds of the present invention which are safe and effective when used in a mammal, and which possess the desired biological activity.

"Solvate" according to the present invention is intended in the conventional sense to mean a complex formed by the combination of a solute (e.g. active compound, salt of active compound) and a solvent (e.g. water). The solvent refers to a solvent known to or easily determined by those skilled in the art. In the case of water, the solvate is generally referred to as a hydrate, such as a hemihydrate, a monohydrate, a dihydrate, a trihydrate, or an alternative amount thereof, and the like.

The in vivo effect of the compound of formula (I) may be exerted in part by one or more metabolites formed in the human or animal body following administration of the compound of formula (I). As mentioned above, the in vivo effects of the compounds of formula (I) may also be exerted via metabolism of the precursor compounds ("prodrugs"). The "prodrug" of the present invention means a compound which is converted into a compound of the present invention by reaction with an enzyme, gastric acid or the like under physiological conditions in an organism, that is, a compound which is converted into a compound of the present invention by oxidation, reduction, hydrolysis or the like of an enzyme and/or a compound which is converted into a compound of the present invention by hydrolysis reaction of gastric acid or the like, or the like.

The "crystal" of the present invention means a solid whose internal structure is formed by repeating constituent atoms (or groups thereof) regularly in three dimensions, unlike an amorphous solid which does not have such a regular internal structure.

The term "pharmaceutical composition" according to the instant invention shall mean a mixture comprising any one of the compounds of the instant invention, including the corresponding isomer, prodrug, solvate, pharmaceutically acceptable salt or chemically protected form thereof, and one or more pharmaceutically acceptable carriers and/or another drug or drugs. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism. The compositions are generally useful in the manufacture of medicaments for the treatment and/or prophylaxis of diseases mediated by one or more kinases.

By "pharmaceutically acceptable carrier" is meant a carrier that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the compound being administered, and that comprises all solvents, diluents or other excipients, dispersants, surfactant isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like. Unless any conventional carrier medium is incompatible with the compounds of the present invention. Some examples of pharmaceutically acceptable carriers that may be used include, but are not limited to, sugars such as lactose, glucose, and sucrose, starches such as corn starch and potato starch, celluloses and their derivatives such as sodium carboxymethyl cellulose, and celluloses and cellulose acetate, malt, gelatin, and the like.

The "excipient" of the present invention refers to an inert substance added to a pharmaceutical composition to further facilitate administration of the compound. Excipients may include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols.

The "EGFR WT" of the present invention is WIDE TYPE EGFR, i.e., wild-type EGFR. The "EGFR (del 19/T790M/C797S)" of the present invention is the EGFR mutated in del 19/T790M/C797S.

Detailed Description

The present invention will be described in further detail with reference to examples, but the present invention is not limited to these examples. The materials used in the examples below are commercially available unless otherwise specified.

Example 1 (R, E) -5 ⁶ - (4-cyclopropylpiperazine-1-carbonyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundecan-3-one

Preparation of step 12- (5-hydroxy-1-methyl-1H-pyrazol-4-yl) -6-isonicotinic acid methyl ester hydrochloride

Methyl 2-chloro-6-methyliisonicotinate (25.0 g,134.7 mmol), 1-methyl-5-hydroxypyrazole (17.2 g,175.1 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (4.9 g,6.7 mmol) and sodium carbonate (28.6 g,269.4 mmol) were added to anisole (600 mL) and stirred under argon at 130℃for 12h. After the completion of the reaction, the reaction system was cooled to room temperature and filtered through celite, and a 4 mol/L1, 4-dioxane solution (67.5 mL,270.0 mmol) of hydrogen chloride was added to the filtrate, and stirred at room temperature for 1 hour, and concentrated and filtered to give the title compound. ESI-MS m/z 248.1[ M+H-HCl ] ⁺.

Step 2 preparation of (R) - (+) -3, 7-dimethyl-6-octenoic acid

(R) - (+) -5-methyl-2- (prop-2-ylene) cyclohexanone (50.0 g,328.4 mmol) was dissolved in 4mol/L hydrogen chloride in 1, 4-dioxane (164.2 mL,656.8 mmol), stirred overnight at 30 ℃, after the reaction was completed, the reaction was added dropwise to 2mol/L potassium hydroxide solution (350.0 mL) and stirred at room temperature for 3h, pH=1 was adjusted with 2.5mol/L dilute hydrochloric acid, the aqueous phase was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give the title compound. ESI-MS m/z 169.2[ M-H ] ^-.

Preparation of tert-amyl (R) - (2, 6-dimethylhept-5-en-1-yl) carbamate

(R) - (+) -3, 7-dimethyl-6-octenoic acid (12.0 g,70.5 mmol), triethylamine (14.3 g,141.0 mmol) were dissolved in toluene (75 mL), stirred at 50℃and then diphenyl azide phosphate (23.3 g,84.6 mmol) was slowly added, stirred at 70℃for 4 hours, t-amyl alcohol (28.0 g,317.3 mmol) was added, stirred at 120℃overnight, after the reaction was completed, the reaction system was cooled to room temperature, quenched by water, extracted with ethyl acetate, dried over anhydrous sodium sulfate and purified by column chromatography to give the title compound. ESI-MS m/z 256.2[ M+H ] ⁺.

Preparation of tert-amyl (R) - (5-hydroxy-2-methylpentyl) carbamate

Tert-amyl (R) - (2, 6-dimethylhept-5-en-1-yl) carbamate (9.5 g,37.2 mmol) is dissolved in methanol (95 mL), the mixture is placed in a precooling mode at a temperature of minus 78 ℃, ozone is introduced into a reaction system, after the raw materials react, argon is introduced into the reaction system for 15min, sodium borohydride (2.8 g,74.4 mmol) is added into the reaction system, the temperature is slowly raised to room temperature, the mixture is stirred overnight, after the reaction is finished, the saturated aqueous solution of ammonium chloride is added for quenching the reaction, most of methanol is evaporated by rotary evaporation, ethyl acetate extraction, anhydrous sodium sulfate drying and column chromatography purification are carried out, and the title compound is obtained. ESI-MS m/z 232.2[ M+H ] ⁺.

Step 5 (R) -5-amino-4-methylpentyl-1-ol hydrochloride preparation

Tert-amyl (R) - (5-hydroxy-2-methylpentyl) carbamate (5.1 g,22.0 mmol) was dissolved in methyl tert-butyl ether (38 mL), 4mol/L hydrogen chloride in 1, 4-dioxane (33.0 mL,132.0 mmol) was added at room temperature, stirred overnight at room temperature, and after the reaction was completed, the title compound was obtained by spin-drying. ESI-MS m/z 118.1[ M+H-HCl ] ⁺.

Step 6 preparation of (R) -5- ((5-bromo-2-nitrophenyl) amino) -4-methylpentan-1-ol

2-Fluoro-4-bromonitrobenzene (1.7 g,7.7 mmol), (R) -5-amino-4-methylpentyl-1-ol hydrochloride (1.3 g,8.5 mmol) and potassium carbonate (3.3 g,23.9 mmol) were added to N, N-dimethylformamide (20 mL), stirred at room temperature overnight, after completion of the reaction, diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate and concentrated to give the title compound. ESI-MS m/z 317.1[ M+H ] ⁺.

Step 7 (R) -5- ((5-bromo-2-nitrophenyl) amino) -4-methylpentylmethylsulfonate preparation

(R) -5- ((5-bromo-2-nitrophenyl) amino) -4-methylpentan-1-ol (2.9 g,9.2 mmol) was dissolved in dichloromethane (25 mL), triethylamine (1.4 g,13.8 mmol) was added, methanesulfonyl chloride (1.3 g,11.0 mmol) was added under ice-bath, stirred at room temperature for 2h, after the reaction was completed, quenched with water, and the organic phase was dried over anhydrous sodium sulfate and concentrated to give the title compound. ESI-MS m/z 395.0[ M+H ] ⁺.

Preparation of methyl 8 (R) -2- (5- ((5- ((5-bromo-2-nitrophenyl) amino) -4-methylpentyl) oxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid

(R) -5- ((5-bromo-2-nitrophenyl) amino) -4-methylpentylmethsulfonate (3.1 g,7.8 mmol), 2- (5-hydroxy-1-methyl-1H-pyrazol-4-yl) -6-isonicotinic acid methyl ester hydrochloride (2.2 g,7.8 mmol) and potassium carbonate (3.2 g,23.4 mmol) were added to N, N-dimethylformamide (25 mL), stirred overnight at 60℃and, after completion of the reaction, diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate and purified by column chromatography to give the title compound. ESI-MS m/z 546.1[ M+H ] ⁺.

Preparation of methyl (R) -2- (5- ((5- ((2-amino-5-bromophenyl) amino) -4-methylpentyl) oxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid

(R) -2- (5- ((5- ((5-bromo-2-nitrophenyl) amino) -4-methylpentyl) oxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid methyl ester (3.4 g,6.2 mmol) was added to ethanol (30 mL), 6mL of water was added, iron powder (3.4 g,62.3 mmol) and ammonium chloride (3.3 g,62.3 mmol) were further added, stirring was carried out at 75℃for 2H, after completion of the reaction, filtration, concentration, dilution with water, extraction with dichloromethane, drying over anhydrous sodium sulfate, concentration gave the title compound. ESI-MS m/z 516.2[ M+H ] ⁺.

Preparation of step 10 (R) -2- (5- ((5- (2-amino-6-bromo-1H-benzo [ d ] imidazol-1-yl) -4-methylpentyl) oxy) -1-methyl-1H-pyrazol-4-yl) -6-isonicotinic acid methyl ester

(R) -2- (5- ((5- ((2-amino-5-bromophenyl) amino) -4-methylpentyl) oxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid methyl ester (3.2 g,6.2 mmol) was dissolved in dichloromethane (30 mL), tert-butanol (6 mL) and cyanogen bromide (0.79 g,7.5 mmol) were added, stirred overnight at 40 ℃, after the completion of the reaction, the reaction was quenched by addition of saturated aqueous sodium bicarbonate solution, the organic layer was separated, dried over anhydrous sodium sulfate, and concentrated to give the title compound. ESI-MS m/z 541.1[ M+H ] ⁺.

Step 11 preparation of (R) -2- (5- ((5- (2-amino-6-bromo-1H-benzo [ d ] imidazol-1-yl) -4-methylpentyl) oxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid

(R) -2- (5- ((5- (2-amino-6-bromo-1H-benzo [ d ] imidazol-1-yl) -4-methylpentyl) oxy) -1-methyl-1H-pyrazol-4-yl) -6-isonicotinic acid methyl ester (3.2 g,5.9 mmol) was dissolved in tetrahydrofuran (30 mL), 1mol/L aqueous sodium hydroxide solution (30 mL) was added, stirred at room temperature for 2 hours, after the reaction was completed, tetrahydrofuran was concentrated and rotary evaporated, pH was adjusted to weak acidity with 4mol/L diluted hydrochloric acid, and the mixture was filtered and dried in vacuo to obtain the title compound. ESI-MS m/z 527.1[ M+H ] ⁺.

Step 12 preparation of (R, E) -5 ⁶ -bromo-1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

(R) -2- (5- ((5- (2-amino-6-bromo-1H-benzo [ d ] imidazol-1-yl) -4-methylpentyl) oxy) -1-methyl-1H-pyrazol-4-yl) -6-methyliisonicotinic acid (2.6 g,4.9 mmol) was dissolved in dichloromethane (60 mL), triethylamine (2.0 g,19.6 mmol) and 2- (1H-benzotrisazo L-1-yl) -1, 3-tetramethylurea tetrafluoroborate (1.9 g,5.9 mmol) were added, stirred at room temperature for 3H, after the reaction was completed, the reaction system was washed three times with water, the organic phase was dried over anhydrous sodium sulfate, and the title compound was obtained by column chromatography purification. ESI-MS m/z 509.1[ M+H ] ⁺.

Preparation of step 13 (R, E) -1 ¹,2⁶, 7-trimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxo-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) pyrazolocyclopentane-5 ⁶ -carboxylic acid

Palladium acetate (0.01 g,0.047 mmol) and 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.027 g,0.047 mmol) were placed in a reaction flask and replaced three times with argon, formic acid (0.254 g,5.5 mmol) and ((R, E) -5 ⁶ -bromo-1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazolo cycloundecan-3-one (0.400 g,0.79 mmol) in N, N-dimethylformamide (10 mL) were added, and after completion of the reaction, the mixture was stirred at 100℃for 3H, filtered and purified by column chromatography to give the title compound ESI-43M 2+M. ⁺.Z.

Step 14 preparation of (R, E) -5 ⁶ - (4-cyclopropylpiperazine-1-carbonyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundecan-3-one

(R, E) -1 ¹,2⁶, 7-trimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxo-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) pyrazolocyclopentane-5 ⁶ -carboxylic acid (0.085 g, 0.178 mmol) was dissolved in dichloromethane (10 mL), triethylamine (0.036 g, 0.178 mmol), 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (0.035 g, 0.178 mmol), 1-hydroxybenzotriazole (0.024 g, 0.178 mmol) and 1-cyclopropylpiperazine (0.022 g, 0.178 mmol) were added sequentially, stirred overnight at room temperature, after the reaction was completed, the dichloromethane was diluted and washed three times with water, dried over anhydrous sodium sulfate, and purified by column chromatography to give the title compound .¹H NMR(400MHz,CDCl₃)δ8.44(s,1H),8.12(s,1H),7.62(s,1H),7.37(s,1H),7.31(d,J＝8.1Hz,1H),7.24(s,1H),4.43(dd,J＝13.0,8.4Hz,1H),4.33(d,J＝13.2Hz,1H),3.90–3.83(m,1H),3.79–3.63(m,6H),3.27–3.16(m,2H),2.86–2.76(m,1H),2.64–2.48(m,6H),2.34–2.27(m,1H),2.19(s,3H),1.96–1.85(m,1H),1.74–1.67(m,1H),1.63(s,1H),1.54–1.43(m,1H),1.20(t,J＝7.2Hz,1H),0.49–0.36(m,4H).ESI-MS m/z:583.3[M+H]⁺.

Example 2 (R, E) -5 ⁶ - (3-hydroxy-3-methylazetidine-1-carbonyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazole cycloundecan-3-one

Preparation method the same as in example 1 except that the starting 1-cyclopropylpiperazine was replaced with 3-methyl-3-acridine hydrochloride to give the title compound .¹H NMR(400MHz,CDCl₃)δ8.42(s,1H),8.12(s,1H),7.65(d,J＝21.1Hz,2H),7.43(d,J＝7.7Hz,1H),7.30(d,J＝6.4Hz,1H),4.47–4.38(m,1H),4.34(d,J＝12.2Hz,2H),4.19(s,3H),3.90–3.81(m,1H),3.81–3.66(m,4H),2.87–2.73(m,1H),2.60(s,3H),2.29–2.18(m,1H),2.14–2.03(m,1H),1.97–1.87(m,1H),1.82–1.74(m,1H),1.59(s,3H),0.89(d,J＝5.0Hz,3H).ESI-MS m/z:544.3[M+H]⁺.

Example 3 (R, E) -1 ¹,2⁶, 7-trimethyl-5 ⁶ - ((((1R, 5S) -8-methyl-3, 8-diazabicyclo [3.2.1] oct-3-yl) methyl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundecan-3-one

Preparation of step 1 (R, E) -1 ¹,2⁶, 7-trimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazolocycloundecane-5 ⁶ -carbaldehyde

Palladium acetate (0.013 g,0.059 mmol), N-butylbis (1-adamantyl) phosphine (0.042 g,0.118 mmol) and sodium formate (0.667 g,9.81 mmol) were placed in a reaction flask and replaced three times with argon, a mixture of (R, E) -5 ⁶ -bromo-1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazolocycloundecan-3-one (0.500 g,0.982 mmol) in N, N-dimethylformamide (10 mL) and formic acid (0.148 mL) and acetic anhydride (0.371 mL) was added, stirred at 30℃for 1H, triethylamine (0.497 g,4.91 mmol) was added, stirred at 110℃for 3H, after the reaction was completed, diluted with ethyl acetate, the aqueous phase was washed with water, and the title compound was dried without aqueous chromatography. ESI-MS m/z 459.2[ M+H ] ⁺.

Step 2 preparation of (R, E) -1 ¹,2⁶, 7-trimethyl-5 ⁶ - ((((1R, 5S) -8-methyl-3, 8-diazabicyclo [3.2.1] oct-3-yl) methyl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundec-3-one

(R, E) -1 ¹,2⁶, 7-trimethyl-3-oxo-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridine-1 (4, 5) -pyrazolocycloundecane-5- ⁶ -carbaldehyde (0.080 g,0.175 mmol), 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride (0.070 g,0.350 mmol) were dissolved in dichloromethane (10 mL), triethylamine (0.071 g,0.70 mmol), glacial acetic acid (0.106 g,1.75 mmol) and sodium triacetoxyborohydride (0.112 g,0.525 mmol) were added in sequence, the reaction was quenched at room temperature overnight, and after the reaction was completed, a saturated aqueous solution of sodium bicarbonate was quenched, diluted with water, the organic phase was dried over anhydrous sodium sulfate, column chromatography was added to purify the title compound .¹H NMR(400MHz,CDCl₃)δ8.65(s,1H),8.48(s,1H),8.15(s,1H),7.66(s,1H),7.31–7.27(m,1H),7.24–7.20(m,1H),4.53–4.42(m,1H),4.36(d,J＝12.9Hz,1H),3.94–3.85(m,1H),3.79(s,3H),3.74–3.59(m,3H),3.48(s,2H),2.88–2.73(m,3H),2.70–2.59(m,5H),2.55(s,3H),2.32–2.22(m,1H),2.15–1.92(m,6H),1.58–1.47(m,1H),0.91(d,J＝6.3Hz,3H).ESI-MS m/z:569.3[M+H]⁺.

Example 4 (R, E) -1 ¹,2⁶, 7-trimethyl-5 ⁶ - (((6-methyl-2, 6-diazaspiro [3.3] hept-2-yl) methyl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 2) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

The procedure was followed except that 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride as the starting material was replaced with 2-methyl-2, 6-diazaspiro [3.3] heptane hydrochloride to prepare the title compound .¹H NMR(400MHz,CDCl₃)δ8.47(d,J＝4.3Hz,2H),8.15(s,1H),7.65(s,1H),7.24(s,1H),7.10(d,J＝7.8Hz,1H),4.50–4.43(m,1H),4.34(d,J＝13.0Hz,1H),3.92–3.85(m,1H),3.82–3.65(m,10H),3.44(s,4H),2.90–2.78(m,1H),2.64(s,3H),2.54(s,3H),2.31–2.20(m,1H),2.17–2.08(m,1H),2.00–1.88(m,1H),1.57–1.46(m,1H),0.90(d,J＝6.0Hz,3H).ESI-MS m/z:555.3[M+H]⁺.

EXAMPLE 5 (R, E) -1 ¹,2⁶, 7-trimethyl-5 ⁶ - (((1S, 4S) -5-methyl-2, 5-diazabicyclo [2.2.1] heptan-2-yl) methyl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazol-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundecan-3-one

The procedure was followed in the same manner as in example 3 except that 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride as the starting material was replaced with (1S, 4S) -2-methyl-2, 5-diazabicyclo [2.2.1] heptane dihydrobromide to give the title compound .¹H NMR(400MHz,CDCl₃)δ8.41(s,1H),8.09(s,1H),7.59(s,1H),7.19(s,2H),7.15(d,J＝6.9Hz,1H),4.45–4.35(m,1H),4.28(d,J＝12.9Hz,1H),3.81(d,J＝13.3Hz,2H),3.74(d,J＝14.4Hz,4H),3.66(d,J＝12.2Hz,1H),3.29(s,2H),3.02(d,J＝8.4Hz,1H),2.79(d,J＝7.4Hz,2H),2.64(d,J＝11.6Hz,1H),2.57(s,3H),2.41(s,3H),2.20(d,J＝6.8Hz,1H),2.04(s,1H),1.92(dd,J＝22.9,5.7Hz,3H),1.76(s,2H),0.82(s,3H).ESI-MS m/z:555.3[M+H]⁺.

Example 6 (R, E) -1 ¹,2⁶, 7-trimethyl-5 ⁶ - ((2-methyl-2, 7-diazaspiro [3.5] nonyl-7-yl) methyl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

The procedure was followed except that 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride as the starting material was replaced with 2-methyl-2, 7-diazaspiro [3.5] nonane dihydrochloride to give the title compound .¹H NMR(400MHz,CDCl₃)δ8.48(s,1H),8.16(s,1H),7.66(s,1H),7.29(s,2H),7.17(d,J＝7.9Hz,1H),4.52–4.42(m,1H),4.35(d,J＝13.1Hz,1H),3.93–3.85(m,1H),3.79(s,3H),3.75–3.69(m,1H),3.55(q,J＝13.7Hz,2H),3.10(s,4H),2.90–2.79(m,1H),2.64(s,3H),2.39(s,3H),2.37–2.26(m,4H),2.19–2.08(m,2H),2.01–1.90(m,2H),1.79(s,4H),0.91(d,J＝6.2Hz,3H).ESI-MS m/z:583.3[M+H]⁺.

Example 7 (R, E) -5 ⁶ - (2-fluoro-5- (4-methylpiperazin-1-yl) phenyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole 2 (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

Step 11 preparation of- (3-bromo-4-fluorophenyl) -4-methylpiperazine

3-Bromo-4-fluoroaniline (2.0 g,10.5 mmol), bis (2-chloroethyl) methylamine hydrochloride (2.0 g,10.5 mmol) and sodium carbonate (3.9 g,36.8 mmol) were placed in a bottle, and isopropanol (190 mL) was added under argon atmosphere and stirred overnight at 100℃and after completion of the reaction, filtered, dried by spin-on, and purified by column chromatography to give the title compound. ESI-MS m/z 273.0[ M+H ] ⁺.

Step 2 preparation of (2-fluoro-5- (4-methylpiperazin-1-yl) phenyl) boronic acid

1- (3-Bromo-4-fluorophenyl) -4-methylpiperazine (1.0 g,3.7 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), pre-cooled under argon protection at-78℃and a tetrahydrofuran solution (2.2 mL) of 2.5mol/L n-butyllithium was added, and stirred for 20min, triisopropyl borate (1.1 g,5.6 mmol) was added, stirred for 1h at-78℃after completion of the reaction, quenched by addition of saturated aqueous ammonium chloride solution, concentrated, extracted with dichloromethane/isopropanol (V ₁:V₂ = 3:1), dried over anhydrous sodium sulfate, and concentrated to give the title compound. ESI-MS m/z 239.1[ M+H ] ⁺.

Step 3 preparation of (R, E) -5 ⁶ - (2-fluoro-5- (4-methylpiperazin-1-yl) phenyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole 2 (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

(R, E) -5 ⁶ -bromo-1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundecan-3-one (0.100 g, 0.197mmol), (2-fluoro-5- (4-methylpiperazin-1-yl) phenyl) boronic acid (0.094 g, 0.3994 mmol), (1, 1' -bis (diphenylphosphino) ferrocene) palladium dichloride (0.015 g, 0.020mmol) and sodium carbonate (0.084 g,0.788 mmol) were added to N, N-dimethylformamide (10 mL), water (2 mL) was added, heated and stirred under argon at 120℃for 6H, after the end of the reaction, the ethyl acetate was diluted with water, washed three times, the organic phase was dried over anhydrous sodium sulfate, and the column was purified to give the title compound by chromatography .¹H NMR(400MHz,CDCl₃)δ8.50(s,1H),8.16(s,1H),7.68(s,1H),7.46(s,1H),7.41(s,2H),7.10(t,J＝9.4Hz,1H),7.02–6.97(m,1H),6.94–6.88(m,1H),4.53–4.44(m,1H),4.39(d,J＝14.1Hz,1H),3.94–3.86(m,1H),3.83–3.72(m,4H),3.21(s,4H),2.91–2.82(m,1H),2.65(s,3H),2.61(s,4H),2.37(s,3H),2.31–2.24(m,1H),2.18–2.10(m,1H),2.00–1.91(m,1H),1.53–1.47(m,1H),0.93(d,J＝6.3Hz,3H).ESI-MS m/z:623.3[M+H]⁺.

Example 8 (R, E) -5 ⁶ - (((4-hydroxy-4- (trifluoromethyl) piperidin-1-yl) methyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazol-2 (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

The procedure was followed except that 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride as the starting material was replaced with 4- (trifluoromethyl) piperidin-4-ol to give the title compound .ESI-MS m/z:612.3[M+H]⁺.¹H NMR(400MHz,DMSO)δ12.56(s,1H),8.43(s,1H),7.93(s,1H),7.57(s,1H),7.54–7.42(m,2H),7.18(d,J＝7.8Hz,1H),5.26(s,1H),4.60(s,2H),4.36(s,1H),4.19(s,1H),4.01(s,1H),3.96–3.82(m,1H),3.74(s,3H),2.82(s,1H),2.56(s,3H),2.22(s,1H),1.98(t,J＝22.8Hz,3H),1.46(s,2H),1.24(s,4H),0.83(d,J＝6.3Hz,5H).

Example 9 (R, E) -1 ¹,2⁶, 7-trimethyl-5 ⁶ - ((4- (oxetan-3-yl) piperazin-1-yl) methyl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazol-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundecan-3-one

The procedure was followed except that 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride as the starting material was replaced with 1- (3-oxetanyl) piperazine to give the title compound .ESI-MS m/z:585.3[M+H]⁺.1H NMR(400MHz,DMSO)δ12.71(s,1H),8.43(s,1H),7.93(s,1H),7.70–7.30(m,3H),7.17(d,J＝7.9Hz,1H),4.54(dd,J＝22.4,16.2Hz,2H),4.38(d,J＝18.5Hz,2H),4.20(d,J＝13.1Hz,1H),3.95(dd,J＝26.5,14.3Hz,2H),3.74(s,3H),3.55(d,J＝26.7Hz,2H),3.39(s,3H),2.56(s,2H),2.42(s,3H),2.26(s,3H),2.03–1.90(m,2H),1.46(s,1H),1.34(s,1H),1.24(s,2H),1.18(d,J＝6.4Hz,1H),0.82(t,J＝11.1Hz,3H).

Example 10 (E) -5 ⁶ - ((4-hydroxy-4- (trifluoromethyl) piperidin-1-yl) methyl) -1 ¹,2⁶ -dimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

Preparation method the procedure was followed in examples 1 and 3, except that the starting material (R) -5-amino-4-methylpentyl-1-ol hydrochloride was replaced with 5-amino-1-pentanol to give the title compound .ESI-MS m/z:598.3[M+H]⁺.1H NMR(400MHz,DMSO)δ12.60(s,1H),8.41(s,1H),8.32(s,1H),7.92(s,1H),7.57(s,1H),7.56–7.43(m,1H),7.19(d,J＝7.9Hz,1H),5.32(s,1H),4.21(s,3H),3.73(s,2H),3.58(s,2H),2.69(d,J＝11.7Hz,2H),2.56(s,2H),2.33–2.18(m,2H),2.02(d,J＝32.6Hz,3H),1.80(s,2H),1.65(t,J＝16.2Hz,3H),1.23(s,4H),0.85(s,1H).

Example 11 preparation of (R, E) -5 ⁶ - ((3-hydroxy-3- (trifluoromethyl) -azetidin-1-yl) methyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazol-2 (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

Preparation method the procedure of example 3 was followed except that the starting 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride was replaced with 3- (trifluoromethyl) azetidine-3-ol hydrochloride to give the title compound .ESI-MS m/z:584.3[M+H]+.¹H NMR(400MHz,DMSO)δ12.72(s,1H),8.42(s,1H),8.18(s,1H),7.93(s,1H),7.53(d,J＝31.3Hz,2H),7.15(d,J＝7.2Hz,1H),5.76(s,1H),4.35(s,1H),4.19(d,J＝12.3Hz,1H),3.95(d,J＝26.6Hz,3H),3.73(s,3H),3.53(d,J＝8.1Hz,2H),3.21(d,J＝6.9Hz,2H),2.78(s,1H),2.55(s,3H),2.20(s,1H),2.10–1.78(m,2H),1.45(s,1H),1.22(s,1H),0.81(s,3H).

Example 12 preparation of (R, E) -5 ⁶ - ((7, 8-dihydro-1, 6-naphthyridin-6 (5H) -yl) methyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

The procedure was followed in the same manner as in example 3 except that 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride as the starting material was replaced with 5,6,7, 8-tetrahydro-1, 6-naphthyridine dihydrochloride to give the title compound .ESI-MS m/z:577.3[M+H]+.¹H NMR(400MHz,DMSO)δ12.71(s,1H),8.39(d,J＝19.9Hz,1H),8.33(s,1H),8.18(s,1H),7.92(s,1H),7.57(s,1H),7.51(d,J＝7.1Hz,1H),7.42(d,J＝6.5Hz,1H),7.24(d,J＝7.1Hz,1H),7.13(s,1H),4.35(s,1H),4.18(d,J＝12.3Hz,1H),4.03–3.88(m,3H),3.78(s,2H),3.72(s,2H),3.61(s,2H),2.87(d,J＝23.4Hz,2H),2.73(d,J＝43.5Hz,3H),2.51(s,3H),2.19(s,1H),2.03–1.82(m,2H),1.44(s,1H),0.80(d,J＝4.2Hz,3H).

Example 13 preparation of (R, E) -1 ¹,2⁶, 7-trimethyl-5 ⁶ - (3-methyl-3- (4-methylpiperazin-1-yl) but-1-yn-1-yl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

(R, E) -5 ⁶ -bromo-1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundecan-3-one (120 mg;0.2358 mmol), 1-methyl-4- (2-methylbutan-3-yn-2-yl) piperazine (53 mg; 0.3536 mmol) was weighed into a 25ml three-necked flask, 6mL N, N-dimethylformamide was dissolved, triphenylphosphine (4 mg; 0.0142 mmol), palladium acetate (2 mg; 0.0071 mmol) was weighed out, and under the protection of argon, 110 ℃ C.) was reacted overnight, after the reaction was complete, 20ml water was added and ethyl acetate was extracted to prepare the title product .ESI-MS m/z:595.3[M+H]⁺.1H NMR(400MHz,DMSO)δ12.84(s,1H),8.42(s,1H),7.93(s,1H),7.73(s,1H),7.57(s,2H),7.28(d,J＝7.8Hz,1H),4.35(s,1H),4.10(dd,J＝36.8,19.1Hz,2H),3.74(s,3H),2.85–2.71(m,3H),2.56(s,3H),2.29(d,J＝24.5Hz,5H),2.04–1.86(m,3H),1.43(s,3H),1.33(d,J＝15.6Hz,2H),1.24(s,6H),0.89–0.78(m,4H).

Example 14 preparation of (R, E) -5 ⁶ - ((3S, 4R) -4-amino-3-methyl-1-oxa-8-azaspiro [4.5] dec-8-yl) methyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundecan-3-one

Preparation method the procedure was followed in example 3 except that starting 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride was replaced with (R) -2-methyl-N- ((3S, 4R) -3-methyl-1-oxo-8-azaspiro [4.5] dec-4-yl) propane-2-sulfinamide to give the title compound ¹H NMR(400MHz,DMSO)δ8.43(s,1H),7.93(s,1H),7.57(s,1H),7.41(dd,J＝53.6,7.2Hz,2H),7.17(d,J＝7.8Hz,1H),4.36(s,1H),4.20(d,J＝12.7Hz,1H),4.03(d,J＝23.0Hz,2H),3.96–3.86(m,1H),3.73(s,3H),3.62(d,J＝8.1Hz,1H),3.55(s,2H),2.99(s,1H),2.87–2.67(m,2H),2.56(s,3H),2.17(dd,J＝18.5,10.8Hz,2H),1.97(dd,J＝24.2,14.9Hz,3H),1.65(dd,J＝24.2,10.3Hz,2H),1.49(d,J＝27.5Hz,3H),1.23(s,6H),1.10(d,J＝5.6Hz,2H),0.81(d,J＝5.2Hz,3H).ESI-MS m/z:613.4[M+H]⁺.

Example 15 preparation of (R, E) -5 ⁶ - (((1R, 5R) -1, 4-diazabicyclo [3.2.1] oct-4-yl) methyl) -1 ¹,2⁶, 7-trimethyl-5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2 (2, 4) -pyridin-1 (4, 5) -pyrazolocycloundec-3-one

The procedure was followed except that 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride as the starting material was replaced with (1R, 5R) -1, 4-diazabicyclo [3.2.1] octane dihydrochloride to give the title compound .¹H NMR(400MHz,CDCl₃)δ8.48(s,1H),8.16(s,1H),7.66(s,1H),7.30(d,J＝6.9Hz,2H),7.18(d,J＝7.9Hz,1H),4.48(dd,J＝13.1,8.3Hz,1H),4.37(d,J＝13.3Hz,1H),3.93–3.87(m,1H),3.79(s,3H),3.73(d,J＝13.1Hz,1H),3.61–3.52(m,2H),3.32(s,1H),3.24–3.15(m,2H),3.12(d,J＝10.9Hz,1H),2.92–2.77(m,4H),2.71–2.67(m,1H),2.64(s,3H),2.47–2.38(m,1H),2.32–2.23(m,1H),2.18–2.08(m,2H),2.03–1.91(m,2H),1.69–1.63(m,1H),0.91(d,J＝6.4Hz,3H).ESI-MS m/z:555.3[M+H]⁺.

Example 16 (R, E) -1 ¹,2⁶, 7-trimethyl-5 ⁶ - ((3-methyl-3, 6-diazabicyclo [3.1.1] hept-6-yl) methyl) -5 ²,5³ -dihydro-1 ¹H,5¹ H-11-oxa-4-aza-5 (2, 1) -benzo [ d ] imidazole-2- (2, 4) -pyridin-1 (4, 5) -pyrazole cycloundecan-3-one

The procedure was followed except that 8-methyl-3, 8-diaza-bicyclo [3.2.1] octane hydrochloride as the starting material was replaced with 3-methyl-3, 6-diazabicyclo [3.1.1] heptane dihydrochloride to prepare the title compound .¹H NMR(400MHz,CDCl₃)δ8.48(s,1H),8.15(s,1H),7.65(s,1H),7.30–7.25(m,2H),7.22(d,J＝7.7Hz,1H),4.51–4.42(m,1H),4.35(d,J＝13.0Hz,1H),3.95–3.85(m,2H),3.85–3.64(m,7H),3.27–2.96(m,4H),2.89–2.70(m,2H),2.64(s,3H),2.57(s,3H),2.30–2.21(m,1H),2.18–2.07(m,2H),2.00–1.90(m,2H),0.93(d,J＝5.9Hz,3H).ESI-MS m/z:555.3[M+H]⁺.

Experimental example 1 evaluation of in vitro cell Activity of Compounds

1. Experimental materials

Test Compound Compounds of the invention prepared in the above examples, were prepared in 10mM stock solution with DMSO and finally diluted to 10 concentrations for detection, and the final concentrations of the compounds for BaF3 (EGFR del 19/T790M/C797S) cell experiments were 1000nM, 250nM, 62.5nM, 15.63nM, 3.91nM, 0.977nM, 0.244nM, 0.0610nM, 0.0153nM, 0.00382nM, and the final concentrations of the compounds for BaF3 (EGFR WT) cell experiments were 50000nM, 12500nM, 3125nM, 781.25nM, 195.31nM, 48.83nM, 12.21nM, 3.05nM, 0.76nM, 0.19nM.

BaF3 (EGFR del 19/T790M/C797S) cells and BaF3 (EGFR WT) cells were all supplied by Kanglong chemical (Beijing) New drug technologies Co., ltd.

Epidermal Growth Factor (EGF) with the product number of invitrogen PHG0311, cellTiter-Glo Luminescent Cell Viability Assay with the product number of Promega G7573, brigatinib with the product number of Selleck S8229.

2. Experimental method

2.1 Cell culture and passage

(1) All cells were cultured in accordance with the methods recommended by ATCC, and the cells were tested in exponential growth phase.

(2) Cell culture medium 1640 medium, 10% FBS,1% Streptomyces lividans, 1% GlutaMax,10ng/mL EGF.

(3) Cell culture conditions of 37℃and 5% CO ₂ and 95% air

(4) The culture medium is changed or passaged every 2-3 days depending on the cell growth.

2.2 Cell plating

Cells were removed from the cell culture flask and resuspended in fresh medium. 30. Mu.L of the cell resuspension was inoculated into 384 well plates, 700 cells/well.

2.3 Administration of drugs

BaF3 (EGFR del 19/T790M/C797S) cells on the basis of the original culture medium (30 mu L), 30nL of drugs with different concentrations are added into the administration group, 30nL of 10mM Brigatinib,DMSO groups are added into the Positive Control (PC) group, 30nL of DMSO is added into the Positive Control (PC) group, two compound holes are arranged in each concentration group, and the culture is continued in a 5% CO ₂ incubator for 3 days. The compounds were formulated by weighing 1-2mg of the compound in advance and preparing 10mM stock solution using DMSO. The drug was diluted with DMSO to a concentration starting at 1mM and starting at the highest concentration, and sequentially diluted to 10 concentration gradients of 1000000nM, 250000nM, 62500nM, 15625nM, 3906.25nM, 9760.563nM, 244.14nM, 61.03nM, 15.26nM, 3.81nM in a 1:3 gradient.

BaF3 (EGFR WT) cells, cells were incubated in an incubator with 5% CO ₂ for 3 days with 150nL of different concentrations of drug in the original medium (30. Mu.L) and 150nL of DMSO in the Positive Control (PC) group and 150nL of 10mM Brigatinib,DMSO groups. The compounds were formulated by weighing 1-2mg of the compound in advance and preparing 10mM stock solution using DMSO. The drug was diluted with DMSO to a concentration of 10mM starting at the highest concentration, and sequentially diluted to 10 concentration gradients of 10000000nM, 2500000nM, 625000nM, 156250nM, 39062.5nM, 9765.63nM, 2441.41nM, 610.35nM, 152.59nM, 38.15nM in a 1:3 gradient.

2.4 Detection

BaF3 (EGFR del 19/T790M/C797S) cells and BaF3 (EGFR WT) cells were equilibrated to room temperature after 3 days of drug treatment by taking CellTiter-Glo Luminescent Cell Viabillity Assay out 30min in advance. Then 30. Mu.L of Celltiter-Glo reagent was added to the Positive Control (PC), dosing and DMSO wells, and shaken. After further incubation for 30min at 5% CO ₂ at 37℃in the dark, the luminescence signal was detected. From the LUM values, the Inhibition (%) =100- (LUM _{Drug delivery hole} –LUM_{PC Hole(s)} )/(LUM _{Solvent control well} -LUM_{PC Hole(s)} ) ×100 for each well was calculated relative to the solvent control wells. According to different drug concentrations and the corresponding inhibition rates, the IC ₅₀ curve is drawn by GraghPad 5.0.0 software, the data are analyzed, the final IC ₅₀ value is obtained, and the experimental results are shown in Table 1.

3. Experimental results

TABLE 1

"-" Indicates no detection.

From the above experimental results, it can be seen that the compounds of the present invention exhibit good inhibitory activity against EGFR-mutated BaF3 cells, while having weak inhibitory activity against EGFR-wild-type BaF3 cells, indicating that the compounds of the present invention have excellent selectivity and are highly promising as EGFR-mutated non-small cell lung cancer therapeutics.

Claims (7)

1. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof, in, Cy-L is selected from ^R1 is selected from _C1-6 alkyl; and ^R2 is hydrogen; ^R3 is selected from _C1-3 alkyl. 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the following compounds: 3. A pharmaceutical composition comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. 4. Use of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 3 in the preparation of a medicament for treating EGFR-mediated diseases. 5. The use according to claim 4, wherein the disease is a tumor disease. 6. The method according to claim 5, wherein the tumor disease is breast cancer, esophageal cancer, bladder cancer, lung cancer, hematopoietic system cancer, lymphoma, medulloblastoma, medulloblastoma, rectal adenocarcinoma, colon cancer, gastric cancer, pancreatic cancer, liver cancer, adenoid cystic carcinoma, prostate cancer, head and neck squamous cell carcinoma, brain cancer, melanoma, oligodendroglioma, glioblastoma, testicular cancer, ovarian clear cell carcinoma, ovarian serous cystadenocarcinoma, thyroid cancer, multiple myeloma or renal cell carcinoma. 7. The use according to claim 5, wherein the tumor disease is bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, hepatocellular carcinoma, mantle cell lymphoma or triple-negative breast cancer.