WO2022083616A1 - Quinazoline compound and pharmaceutical composition thereof - Google Patents

Abstract

A novel compound with cancer therapeutic activity. The present invention also relates to a method for preparing these compounds and a pharmaceutical composition containing same. (I)

Description

A kind of quinazoline compound and pharmaceutical composition thereof technical field

The present invention relates to a novel compound having cancer therapeutic activity. The present invention also relates to methods for the preparation of these compounds and pharmaceutical compositions containing them.

technical background

Clinical data show that RAS is the gene with the highest mutation rate in human tumors, about 20-30% of all tumors have RAS mutations, about 98% of pancreatic cancer, 52% of colon cancer, 43% of multiple myeloma, And 32% of lung adenocarcinomas have RAS gene mutations. The most common mutation of RAS is point mutation, which often occurs at codons 12, 13, and 61, of which the 12th codon mutation is the most common. KRAS-G12C mutations account for approximately 10-20% of KRAS mutations and 14% in non-small cell lung cancer.

For more than 40 years, scientists have been trying to develop drugs that target KRAS. AMG510 developed by AMGEN is the first KRAS G12C inhibitor to enter the clinic. In December 2020, AMGEN submitted a New Drug Application for AMG510 to the US FDA for the treatment of KRAS G12C-mutated advanced metastatic non-small cell lung cancer. The FDA has previously granted it Breakthrough Therapy designation. In May 2021, the FDA approved the world's first KRAS-targeted drug, AMG510.

ARAXES company patent WO2015054572A1 describes a KRAS inhibitor with a specific axial chiral fragment in the form of a racemate. The molecule theoretically has both R/S chirality, but the patent does not describe which is the dominant active configuration.

Jinfang Company's patent WO2020177629A1 discloses the activity data of specific chiral molecules (Table 1). According to the description, the two chiral molecules Z2-1/Z2-2 and Z25-1/Z25-2 with the same axial chiral fragment, the more active molecules Z2-1 and Z25-2 have completely opposite configurations.

Table 1

Compound number	H358IC 50 (μM)	A549IC 50 (μM)
Z2	0.016	3.266
Z2-2	0.023	2.165
Z25	0.079	2.771
Z25-2	0.044	3.424

SUMMARY OF THE INVENTION

The present invention provides a chiral KRAS inhibitor with a novel structure. Unexpectedly, this chiral molecule is more active than its corresponding axial chiral isomer, with a greater than 100-fold difference in activity. The use of chiral molecules with better activity can effectively reduce the dosage of drugs and avoid adverse drug reactions, which has important clinical significance.

The present invention provides a chiral compound, a deuterated compound or a pharmaceutically acceptable salt represented by the general formula (I),

wherein X is selected from C _3-14 cycloalkyl, 3-14 membered heterocyclyl, 3-14 membered heterocyclylamino; X is optionally further substituted with 1-4 substituents, wherein each substituent is independently Selected from C _1-6 alkyl, amino, C _1-6 aminoalkyl, carbamoyl, C _1-6 carbamoyl alkyl, carboxyl, C _1-6 carboxyalkyl, cyano, C _1-6 cyanoalkyl, halogen, C _1-6 haloalkyl, hydroxy, C _1-6 hydroxyalkyl, said C _1-6 alkyl optionally being selected from one or more of amino, carbamoyl, carboxyl , cyano, halogen, hydroxyl, oxo substituted;

R ₁ is acryloyl, substituted acryloyl or

R ₂ is selected from H, C _2-6 alkenyl, C _1-6 alkoxy, C _1-6 haloalkoxy, C _1-6 alkyl, C _1-6 alkane substituted by 4-10-membered heterocyclyl base, C _1-6 alkylsulfonyl, C _1-6 alkylsulfinyl, C _1-6 alkylthio, C _2-6 alkynyl, C _1-6 alkylamino, amino, C _1-6 Aminoalkyl, carbamoyl, C _1-6 carbamoylalkyl, C _1-6 carboxyalkyl, cyano, C _1-6 cyanoalkyl, halogen, C _1-6 haloalkyl, substituted or unsubstituted substituted aryl, substituted or unsubstituted 5-10 membered heteroaryl, substituted or unsubstituted C _3-7 cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclyl, hydroxy or oxo, The C _1-6 alkyl group may be optionally substituted by one or more groups selected from amino, carbamoyl, carboxyl, cyano, halogen, hydroxyl, and oxo;

R ₃ is selected from H, halogen, C _1-6 alkyl, substituted C _1-6 alkyl, C _2-6 alkenyl, substituted C _2-6 alkenyl, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl;

R ₄ or R ₅ is independently selected from H, halogen, C _1-6 alkyl, substituted C _1-6 alkyl, C _3-6 cycloalkyl, substituted C _3-6 cycloalkyl;

R ₆ is selected from hydroxyl, oxo, C _1-6 alkoxy, C _3-10 cycloalkyloxy, cyano-substituted cyclopropyl C _1-6 alkyleneoxy, the C _{1- 6} alkoxy is optionally further substituted by one or more substituents selected from halogen, hydroxy, C _1-6 alkoxy, C _3-8 cycloalkyl;

m or n are independently selected from 0, 1, 2, 3, 4;

Y is selected from a 3-14 membered heterocyclyl group directly connected to the quinazoline ring through a C atom on the ring, and the heterocyclyl group is optionally further selected from 1-4 members R ₇ or R ₈ is substituted; R ₇ or R ₈ is independently selected from H, =O, =S, cyano, halogen, nitro, C _1-6 alkyl, -C _0-6 alkylene-OR _a , -C _0-6 alkylene-OC(O)N(R _a ) ₂ , -C _0-6 alkylene-N(R _a ) ₂ , -C _0-6 alkylene-NR _a C( O)R _a , -C _0-6 alkylene-NR _a C(O)N(R _a ) ₂ , -C _0-6 alkylene-NR _a S(O)R _a , -C _0-6 Alkylene-NR _a S(O) ₂ R _a , -C _0-6 alkylene-S(=O)R _a , -C _0-6 alkylene-S(=O) ₂ R _a , - C _0-6 alkylene-SR _a , -C _0-6 alkylene-S(R _a ) ₅ , -C _0-6 alkylene-C(=O)R _a , -C _0-6 alkylene Alkyl-C(=O)OR _a , -C _0-3 alkylene-C(=O)N(R _a ) ₂ , C _2-6 alkenyl, C _2-6 alkynyl, acryloyl, - C _0-6 alkylene-C _3-14 cycloalkyl, -C _0-6 alkylene-(3-14-membered heterocyclic group), -C _0-6 alkylene-C _6-14 aryl Or -C _0-6 alkylene-(5-14-membered heteroaryl), the C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, -C _0-6 alkylene base-C _3-14 cycloalkyl, -C _0-6 alkylene-(3-14-membered heterocyclic group), -C _0-6 alkylene-C _6-14 aryl or -C _0-6 Alkylene-(5-14 membered heteroaryl) may optionally be substituted with one or more R _a ; each R _a is independently selected from H, halogen, hydroxy, amino, oxo, Nitro, cyano, carboxyl, C _1-6 alkyl, C _1-6 hydroxyalkyl, C _1-6 aminoalkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _1-6 Haloalkoxy, C _1-6 heteroalkyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, C _6-14 aryl, 5-14 membered heteroaryl, C _1-6 acyl or

In some embodiments, X in formula (I) is selected from a 4-10 membered heterocyclyl group optionally further selected from 1-4 members selected from C _1-6 alkyl, cyano substituted by substituents of C _1-6 cyanoalkyl, C _1-6 hydroxyalkyl; preferably

more preferably

In some embodiments, R in formula ( ^I ) is selected from

preferably

In some embodiments, R ² in formula (I) is selected from H, halogen or C _1-3 alkyl; preferably H.

In some embodiments, ^R in formula (I) is selected from H, C _1-6 alkyl, substituted C _1-6 alkyl, C _2-6 alkenyl, substituted C _2-6 alkenyl, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl; preferably C _1-3 alkyl, C _3-6 cycloalkyl or C _2-4 alkenyl; more preferably ethyl, vinyl or cyclo propyl.

In some embodiments, R ⁴ or R ⁵ in formula (I) is selected from H, halogen or C _1-6 alkyl; preferably R ⁴ is H and R ⁵ is methyl.

In some embodiments, R in formula (I) is selected from C ^1-6 alkoxy, C _3-8 cycloalkyloxy, cyano-substituted cyclopropyl C _{1-6 alkyleneoxy} , The C _1-6 alkoxy group is optionally further substituted by one or more substituents selected from halogen, hydroxyl, methoxy, C _3-8 cycloalkyl; preferably R ⁶ is selected from

In some embodiments, Y in formula (I) is selected from

preferably

more preferably

In some embodiments, R ₇ in formula (I) is selected from H, cyano, halogen, C _1-6 alkyl, -C _0-6 alkylene-OR _a , -C _0-6 alkylene- OC(O)N(R _a ) ₂ , -C _0-6 alkylene-N(R _a ) ₂ , -C _0-6 alkylene-NR _a C(O)R _a , -C _0-6 Alkylene-NR _a C(O)N(R _a ) ₂ , -C _0-6 alkylene-NR _a S(O)R _a , -C _0-6 alkylene-NR _a S(O) ₂ R _a , -C _0-6 alkylene-S(=O)R _a , -C _0-6 alkylene-S(=O) ₂ R _a , -C _0-6 alkylene-SR _a , -C _0-6 alkylene-S(R _a ) ₅ , -C _0-6 alkylene-C(=O)R _a , -C _0-6 alkylene-C(=O)OR _a , -C _0-3 alkylene-C(=O)N(R _a ) ₂ , acryloyl, -C _0-6 alkylene-C _3-14 cycloalkyl, -C _0-6 alkylene -(3-14 membered heterocyclyl), -C _0-6 alkylene-C _6-14 aryl or -C _0-6 alkylene-(5-14 membered heteroaryl), the C _{1 -6} alkyl, -C _0-6 alkylene-C _3-14 cycloalkyl, -C _0-6 alkylene-(3-14 membered heterocyclyl), -C _0-6 alkylene- C _6-14 aryl or -C _0-6 alkylene-(5-14-membered heteroaryl) optionally can also be substituted by 1 or more R _a ; each R _a is independently selected from H, halogen, hydroxyl, amino, oxo, cyano, carboxyl, C _1-6 alkyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, C _6-14 aryl, 5-14 Member heteroaryl, C _1-6 acyl or

Preferably R ₇ is selected from H, acryloyl, C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 acyl, -C _0-3 alkylene-C _1-6 alkoxy, C _{1- 6} hydroxyalkyl, C _1-6 cyanoalkyl, -C _0-3 alkylene-S(=O) ₂ -C _1-6 alkyl, C _3-6 keto,

-C _0-3 alkylene-C _3-6 cycloalkyl, -C _0-3 alkylene-C _3-8 heterocyclyl, -C _0-3 alkylene-phenyl, -C _{0- 3} alkylene-pyrazolyl, the -C _0-3 alkylene-C _3-6 cycloalkyl, -C _0-3 alkylene-C _3-8 heterocyclyl, -C _0-3 Alkylene-phenyl, -C _0-3 alkylene-pyrazolyl are optionally further selected from one or more groups selected from C _1-6 acyl, hydroxy, C _1-6 alkyl, C _3-6 ring Alkyl, halogen,

or cyano substituent, the C _3-8 heterocyclic group contains a heteroatom selected from O or N; more preferably R ₇ is selected from H, C _1-6 alkyl, acetyl,

In some embodiments, R ₈ in formula (I) is selected from H or C _1-6 alkyl; preferably H.

In some embodiments, the chiral compound represented by general formula (I) is selected from compounds of formula (IA)-(ID),

; preferably

Wherein, the substituents are as defined in the general formula (I).

The present invention also provides a chiral compound, a deuterated substance or a pharmaceutically acceptable salt, which is selected from:

preferably

The present invention provides a pharmaceutical composition, the pharmaceutical composition contains the chiral compound, deuterated compound or pharmaceutically acceptable salt of any one of formula (I) and at least one pharmaceutically acceptable excipient, and the pharmaceutical composition The content of the chiral compound is higher than that of its corresponding axial chiral isomer.

The present invention also provides a pharmaceutical composition, in which the content of the chiral compound, deuterated compound and pharmaceutically acceptable salt represented by the general formula (I) in the active ingredient is higher than 51%, 61% and 71% , 81%, 91%, 99% or higher.

The present invention also provides a pharmaceutical composition, wherein the therapeutically effective amount of at least one compound represented by formula (I), a deuterated substance, a pharmaceutically acceptable salt and a pharmaceutically acceptable adjuvant in the pharmaceutical composition is 0.0001:1 by mass percentage -10.

The present invention also provides the use of the chiral compound, deuterated compound, pharmaceutically acceptable salt or pharmaceutical composition containing the compound represented by structural formula (I) in the preparation of medicine; preferably, the medicine is an anticancer medicine.

Preferably, the application is the application of a medicament for the treatment of a disease mediated by KRAS G12C. Preferably, the disease is cancer.

Preferably, the cancer is selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, small cell lung cancer, pleomorphic lung cancer, ovarian cancer, esophagus Carcinoma, melanoma, colorectal cancer, hepatoma, head and neck tumor, hepatocholangiocarcinoma, myelodysplastic syndrome, glioblastoma, prostate cancer, thyroid cancer, Schwann cell tumor, lung squamous cell Carcinoma, lichenoid keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer, or liposarcoma.

The present invention also provides a method for treating and/or preventing cancer, comprising administering a therapeutically effective amount of a chiral compound, a deuterated compound, a pharmaceutically acceptable salt of the general formula (I), or a chiral compound containing the compound represented by the structural formula (I) to a subject to be treated. Pharmaceutical composition; preferably the cancer is mediated by a KRAS G12C, HRAS G12C or NRAS G12C mutation.

Preferably, in the above method, the cancer is selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, small cell lung cancer, pleomorphic Lung cancer, ovarian cancer, esophageal cancer, melanoma, colorectal cancer, hepatoma, head and neck tumors, hepatocholangiocarcinoma, myelodysplastic syndrome, glioblastoma, prostate cancer, thyroid cancer, Schwann cells tumor, lung squamous cell carcinoma, lichenoid keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer or liposarcoma.

Unless otherwise indicated, general chemical terms used in the structural formulae have their ordinary meanings.

For example, unless otherwise specified, the term "halogen" as used herein refers to fluorine, chlorine, bromine or iodine.

In the present invention, unless otherwise specified, "alkyl" includes linear or branched monovalent saturated hydrocarbon groups. For example, alkyl includes methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 3-(2-methyl)butyl, 2 -pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl, 2-methylpentyl, etc. Similarly, " _1-8 " in "C _1-8 alkyl" refers to a group containing 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms arranged in straight or branched chain form group.

"C _1-3 alkylene" means a divalent saturated hydrocarbon group including straight or branched chains. For example methylene, 1,2-ethylene, 1,3-propylene or 1,2-isopropylene. Similarly, " _1-6 " in " _C1-6 alkylene" refers to groups containing 1, 2, 3, 4, 5 or 6 carbon atoms arranged in straight or branched chain form.

In the present invention, "a", "an", "the", "at least one" and "one or more" are used interchangeably. Thus, for example, a composition comprising "a" pharmaceutically acceptable excipient can be interpreted to mean that the composition includes "one or more" pharmaceutically acceptable excipients.

The term "aryl", in the present invention, unless otherwise specified, refers to an unsubstituted or substituted mono- or fused-ring aromatic group comprising atoms of a carbocyclic ring, having a fully conjugated pi-electron system. Preferably, the aryl group is a 6-14 membered monocyclic or polycyclic aromatic ring group. Preferred are phenyl and naphthyl. Most preferred is phenyl. The aryl ring can be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring.

The term "heterocyclyl", in the present invention, unless otherwise specified, refers to an unsubstituted or substituted 3-14 membered stable ring consisting of carbon atoms and 1-3 heteroatoms selected from N, O or S. system, which is a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, such as a spiro or bridged ring. Nitrogen or sulfur heteroatoms in heterocyclyl groups can be selectively oxidized, and nitrogen heteroatoms can be selectively quaternized. The heterocyclyl group can be attached to any heteroatom or carbon atom to form a stable structure. Examples of such heterocyclyl groups include, but are not limited to, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidyl, tetrahydrofuranyl, dioxolanyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone and tetrahydro oxadiazolyl. The heterocyclyl group can be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl group.

The term "spirocyclyl", in the present invention, unless otherwise specified, refers to a polycyclic compound in which two monocyclic rings share one carbon atom, and examples of these spirocyclyls include but are not limited to

The term "bridged ring group", in the present invention, unless otherwise specified, refers to a polycyclic compound in which two monocyclic rings share two or more carbon atoms, and examples of these bridged ring groups include but are not limited to

The term "heteroaryl", in the present invention, unless otherwise specified, refers to an unsubstituted or substituted stable 5- or 6-membered monocyclic aromatic ring system or an unsubstituted or substituted 9-14 membered benzo-fused A heteroaromatic ring system or a polycyclic heteroaromatic ring system consisting of carbon atoms and 1-4 heteroatoms selected from N, O or S, and wherein the nitrogen or sulfur heteroatoms may be optionally replaced by oxidation, the nitrogen heteroatom can be selectively quaternized. Heteroaryl groups can be attached to any heteroatom or carbon atom to form a stable structure. Examples of heteroaryl groups include, but are not limited to, thienyl, furyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiazolyl, thiadiazolyl, triazolyl, pyridyl, pyridyl Azinyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, benzofuranyl, benzothienyl, benzisoxazolyl, benzothiazolyl, benzothiazolyl, benzene thiadiazolyl, benzotriazolyl adenine, quinolinyl or isoquinolinyl. The heteroaryl group can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is a heteroaryl ring.

The term "cycloalkyl" refers to a cyclic saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent having 3 to 14 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl . The cycloalkyl group can be fused to an aryl, heterocyclyl or heteroaryl ring, wherein the ring attached to the parent structure is a cycloalkyl group.

The term "substituted" refers to the replacement of one or more hydrogen atoms in a group with the same or a different substituent, respectively. Typical substituents include, but are not limited to, H, =O, =S, cyano, halogen, nitro, C _1-6 alkyl, -C _0-6 alkylene -OR _a , -C _0-6 alkylene base-OC(O)N(R _a ) ₂ , -C _0-6 alkylene-N(R _a ) ₂ , -C _0-6 alkylene-NR _a C(O)R _a , -C _{0 -6} alkylene-NR _a C(O)N(R _a ) ₂ , -C _0-6 alkylene-NR _a S(O)R _a , -C _0-6 alkylene-NR _a S( O) ₂ R _a , -C _0-6 alkylene-S(=O)R _a , -C _0-6 alkylene-S(=O) ₂ R _a , -C _0-6 alkylene- SR _a , -C _0-6 alkylene-S(R _a ) ₅ , -C _0-6 alkylene-C(=O)R _a , -C _0-6 alkylene-C(=O) OR _a , -C _0-3 alkylene-C(=O)N(R _a ) ₂ , C _2-6 alkenyl, C _2-6 alkynyl, acryloyl, -C _0-6 alkylene- C _3-14 cycloalkyl, -C _0-6 alkylene-(3-14-membered heterocyclic group), -C _0-6 alkylene-C _6-14 aryl or -C _0-6 alkylene base-(5-14 membered heteroaryl), the C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, -C _0-6 alkylene-C _3-14 cycloalkane base, -C _0-6 alkylene-(3-14 membered heterocyclic group), -C _0-6 alkylene-C _6-14 aryl or -C _0-6 alkylene-(5-14 heteroaryl) optionally can also be substituted by one or more R _a ; each R _a is independently selected from H, halogen, hydroxyl, amino, oxo, nitro, cyano, carboxyl, C _1-6 alkyl, C _1-6 hydroxyalkyl, C _1-6 aminoalkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _1-6 haloalkoxy, C _1-6 Heteroalkyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, C _6-14 membered aryl, 5-14 membered heteroaryl, C _1-6 membered acyl or

In some embodiments, the substituents are independently selected from the group consisting of -F, -Cl, -Br, -I, -OH, trifluoromethoxy, ethoxy, propoxy, isopropoxy, n-butoxy group, isobutoxy, tert-butoxy, -SCH ₃ , -SC ₂ H ₅ , carboxaldehyde, -C(OCH ₃ ), cyano, nitro, -CF ₃ , -OCF ₃ , amino, dimethyl amino, methylthio, sulfonyl and acetyl groups.

Examples of substituted alkyl groups include, but are not limited to, 2,3-dihydroxypropyl, 2-aminoethyl, 2-hydroxyethyl, pentachloroethyl, trifluoromethyl, methoxymethyl, pentafluoroethyl , phenylmethyl, dioxinylmethyl and piperazinylmethyl.

Examples of substituted alkoxy groups include, but are not limited to, 2-hydroxyethoxy, 2-fluoroethoxy, 2,2-difluoroethoxy, 2-methoxyethoxy, 2-aminoethoxy, 2,3-dihydroxypropoxy, cyclopropylmethoxy, aminomethoxy, trifluoromethoxy, 2-diethylaminoethoxy, 2-ethoxycarbonylethoxy, 3- Hydroxypropoxy.

The term "hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxy groups, eg, -(alkylene)-OH.

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, eg, -(alkylene)-halogen.

The term "alkoxy" refers to -O-(alkyl).

The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids.

Since the compound represented by formula (I) will be used as a medicine, it is preferable to use a certain purity, for example, at least 60% pure, more suitably at least 75% pure, particularly suitably at least 98% pure (% by weight) Compare).

Prodrugs of the compounds of the present invention are included within the scope of the present invention. In general, the prodrugs refer to functional derivatives that are readily converted into the desired compound in vivo. For example, any pharmaceutically acceptable salt, ester, salt of ester or other derivative of a compound of the present application which, upon administration to a recipient, is capable of providing, directly or indirectly, a compound of the present application or a pharmaceutically active metabolite thereof or Residues. Particularly preferred derivatives or prodrugs are those that increase the bioavailability of the compounds of the present application when administered to a patient (eg, make orally administered compounds more readily absorbed into the blood), or promote the delivery of the parent compound to biological organs or Those compounds that are delivered to the site of action (eg, the brain or lymphatic system). Therefore, the term "administration" in the treatment methods provided by the present invention refers to the administration of the compounds disclosed in the present invention that can treat different diseases, or, although not explicitly disclosed, can be transformed into the disclosed compounds in vivo after administration to a subject compounds of compounds.

Obviously, the definition of any substituent or variable at a particular position in a molecule is independent of other positions in the molecule. It is easy to understand that those skilled in the art can select the substituents or substituted forms of the compounds of the present invention by means of the prior art and the methods described in the present invention, so as to obtain chemically stable and easy-to-synthesize compounds.

The compounds of the present invention may contain one or more asymmetric centers and may thereby give rise to diastereomers and optical isomers. The present invention includes all possible diastereomers and racemic mixtures thereof, substantially pure resolved enantiomers thereof, all possible geometric isomers and pharmaceutically acceptable salts thereof.

When the compound represented by formula (I) has tautomers, unless otherwise stated, the present invention includes any possible tautomers and pharmaceutically acceptable salts thereof, and mixtures thereof.

Substitution of compounds of formula (I) with heavier isotopes (eg deuterium) may provide certain therapeutic advantages due to greater metabolic stability, eg increased in vivo half-life or reduced dosage requirements.

When the compounds of formula (I) and their pharmaceutically acceptable salts exist as solvates or polymorphs, the present invention includes any possible solvates and polymorphs. The type of solvent that forms the solvate is not particularly limited as long as the solvent is pharmaceutically acceptable. For example, water, ethanol, propanol, acetone and similar solvents can be used.

The term "composition", in the present invention, refers to a product comprising a specified amount of each of the specified ingredients, as well as any product produced directly or indirectly from a combination of the specified amounts of each of the specified ingredients. Accordingly, pharmaceutical compositions containing the compounds of the present invention as active ingredients and methods of preparing the compounds of the present invention are also part of the present invention. In addition, some of the crystalline forms of the compounds may exist as polymorphs, and such polymorphs are included in the present invention. In addition, some of the compounds may form solvates with water (ie, hydrates) or common organic solvents, and such solvates are also within the scope of this invention.

The pharmaceutical composition provided by the present invention comprises a compound represented by formula (I) (or a pharmaceutically acceptable salt thereof) as an active component, a pharmaceutically acceptable excipient and other optional therapeutic components or Accessories. Although in any given situation, the most appropriate mode of administration of the active ingredient will depend on the particular subject to be administered, the nature of the subject and the severity of the condition. The pharmaceutical compositions of the present invention may conveniently be presented in unit dosage form and prepared by any of the methods of preparation well known in the art of pharmacy.

The pharmaceutical composition of the present invention comprises a pharmaceutically acceptable carrier and a compound represented by formula (I) or its stereoisomer, tautomer, polymorph, solvate, pharmaceutically acceptable salt thereof, its prodrugs. The compound represented by formula (I) or a pharmaceutically acceptable salt thereof, combined with one or more other compounds with therapeutic activity are also included in the pharmaceutical composition of the present invention.

The pharmaceutical compositions of the present invention include pharmaceutical compositions suitable for oral, rectal, topical and parenteral (including subcutaneous, intramuscular, intravenous) administration. Tablets or capsules containing a compound or pharmaceutical composition of the present invention may contain one or more accessory ingredients. Preferably, each tablet or capsule contains about 0.05 mg to 5 g of active ingredient. The present invention also provides pharmaceutical compositions suitable for injection, including sterile aqueous solutions or dispersions. Further, the above-mentioned pharmaceutical compositions can be prepared in sterile powder form for the extemporaneous preparation of sterile injectable solutions or dispersions. The pharmaceutical compositions provided by the present invention may be in a form suitable for topical administration, eg, an aerosol, cream, ointment, lotion, dusting powder, or other similar dosage forms. Further, the pharmaceutical composition provided by the present invention can be in a form suitable for use in a transdermal drug delivery device. The pharmaceutical composition provided by the present invention can be in the form of rectal administration by using a solid as a carrier. Unit-dose suppositories are the most typical dosage form. Suppositories can be conveniently prepared by first mixing the pharmaceutical composition with softened or melted excipients, then cooling and moulding.

The formulations described above may also include, as appropriate, one or more additional adjuvant components such as diluents, buffers, flavoring agents, binders, surfactants, thickeners, lubricants and preservatives (including antioxidants) etc. Further, other adjuvants may also include osmotic enhancers that adjust the isotonic pressure of the drug and blood. The pharmaceutical composition comprising the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, can be prepared in the form of a powder or a concentrated solution.

In general, to treat the conditions or disorders indicated above, the dosage level of the drug is about 0.01 mg/kg body weight to 150 mg/kg body weight per day, or 0.5 mg to 7 g per patient per day. It will be appreciated, however, that lower or higher doses than those described above may be required. The specific dosage level and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, route of administration, excretion rate, drug combination condition and the severity of the specific disease being treated.

Description of drawings

Fig. 1 is the X-ray single crystal diffraction pattern A of compound 1a prepared in Example 1A.

Fig. 2 is the X-ray single crystal diffraction pattern B of compound 1a prepared in Example 1A.

Figure 3 is an absolute configuration diagram of compound 1a prepared in Example 1A.

Figure 4 is an in vivo pharmacodynamic study A on the human pancreatic cancer MIA PaCa-2 CDX tumor model.

Figure 5 is an in vivo pharmacodynamic study B on the human pancreatic cancer MIA PaCa-2 CDX tumor model.

Figure 6 is an in vivo pharmacodynamic study of a human lung cancer PDX tumor model.

Detailed ways

In order to make the above content clearer and clearer, the present invention will further illustrate the technical solutions of the present invention with the following examples. The following examples are only used to illustrate the specific embodiments of the present invention, so that those skilled in the art can understand the present invention, but are not intended to limit the protection scope of the present invention. In the specific embodiments of the present invention, the technical means or methods that are not specifically described are conventional technical means or methods in the field.

All parts and percentages herein are by weight and all temperatures are in degrees Celsius unless otherwise indicated. The following abbreviations are used in the examples:

BOP: Carter condensing agent;

CDI: carbonyldiimidazole;

DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene;

DIEA: N,N-diisopropylethylamine;

DMF: N,N-dimethylformamide;

DCM: dichloromethane;

Dioxane: Dioxane;

ESI-MS: Electrospray Ionization Mass Spectrometry;

EtOH: ethanol;

HOAc: glacial acetic acid;

MeOH: methanol;

NIS: N-iodosuccinimide;

NCS: N-chlorosuccinimide;

PE:EA: the ratio of petroleum ether and ethyl acetate;

POCl ₃ : phosphorus oxychloride;

SOCl ₂ : thionyl chloride;

THF: tetrahydrofuran;

TFA: trifluoroacetic acid;

TEA: triethylamine;

Toluene: toluene;

Sphos Pd G2: Chloro(2-dicyclohexylphosphino-2,6-dimethoxy-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II) );

Pd(dppf)Cl ₂ .CH ₂ Cl ₂ : [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex;

Pd(PPh ₃ ) ₄ : tetrakis(triphenylphosphine)palladium;

Pre-TLC: Thin-layer chromatography on silica gel plates.

Synthesis of intermediate compound M1

Step 1: Synthesis of Compound M1-2

To a mixture of compound M1-1 (40 g), HOAc (76.8 g), EtOH (400 mL) and _H2O (160 mL) was added iron powder (26.52 g) portionwise at room temperature. The resulting mixture was stirred at room temperature for 2 hours and then neutralized with NaOH (5N) solution. The mixture was then extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo to give the desired crude product as a brown oil (34 g, 98% yield), compound M1-2.

ESI-MS m/z: 190.02 [M+H] ⁺ .

Step 2: Synthesis of Compound M1-3

2,2,2-Trichloroethane-1,1-diol (66.4 g) and Na ₂ SO ₄ (503.4 g) were dissolved in water (560 mL) and then heated to 55°C. Water (240 mL) and 35% HCl (72 mL) containing compound M1-2 (34 g) were added, followed by an aqueous solution (100 mL) of hydroxylamine hydrochloride (81.4 g). The resulting mixture was stirred at 90°C for 3 hours and a yellow precipitate formed. The mixture was cooled to room temperature. The solid was collected by filtration, rinsed with water, and air-dried to give the product as a tan solid (47 g, 99% yield), compound M1-3.

ESI-MS m/z: 261.03 [M+H] ⁺ .

Step 3: Synthesis of Compounds M1-4

Compound M1-3 (47 g) was added to concentrated sulfuric acid (300 mL) at 60 °C, the temperature was raised to 90 °C and maintained for 3 hours, the reaction was complete, the reaction mixture was cooled to room temperature and poured into ice water. The yellow precipitate was collected by filtration and dried to give the product as a black solid (43 g, 99% yield), compound M1-4.

Step 4: Synthesis of Compounds M1-5

To a solution of compound M1-4 (43 g) in _NaOH (2N, 500 mL) at ₀ °C was added H2O2 solution (30%, 80 mL) and the resulting mixture was stirred at 0 °C for 30 min. Then moved to room temperature and stirred for 2 hours, the reaction was complete, the mixture was poured into ice water and then acidified with concentrated HCl solution, the precipitate was collected by filtration and air-dried to obtain a white solid product (20 g, 48.9% yield), that is, compound M1-5.

ESI-MS m/z: 233.97 [M+H] ⁺ .

Step 5: Synthesis of Compounds M1-6

To a solution of compound M1-5 (20 g) in DMF (200 mL) was added NIS (29 g) at room temperature and the resulting mixture was stirred at 70 °C overnight. The reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo to give the desired crude product (30 g, 98% yield) as a brown solid, i.e. Compounds M1-6.

ESI-MS m/z: 359.87 [M+H] ⁺ .

1H NMR (500 MHz, DMSO) δ 13.34 (s, 1H), 7.99 (s, 1H), 6.87 (s, 2H).

Step 6: Synthesis of Compound M1

At room temperature, bis(imidazol-1-yl)methanone (2.70 g) was added to crude compound M1-6 (4.0 g) in THF (20 mL), followed by N-ethyl-N-isopropylpropane -2-amine (1.44g, 1.94mL) was added to it, the mixture was moved to 50°C for reaction, and the reaction was about 2 hours. Compound M1-6 was almost completely converted into an intermediate product, and then the mixture was added dropwise to iced ammonia water (35mL) , the reaction was completed after stirring for 5 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, the residue was purified by flash silica gel column chromatography (petroleum ether/ethyl acetate = 70:30) , the desired target product compound M1 (1.64 g) was obtained as a brown solid.

Synthesis of intermediate compound M2

Step 1: Synthesis of Compound M2-1

At room temperature, compound M1-5 (3.6 g) was dissolved in DMF (20 mL), NCS (2.05 g) was added, and the reaction was carried out at 70° C. for 1.5 h. The reaction was completed, cooled to room temperature, diluted with water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered and spin-dried to obtain a brown solid M2-1 (3.86 g, crude product).

ESI-MS m/z: 267.91 [M+H] ⁺ .

Step 2: Synthesis of Compound M2-2

At room temperature, compound M2-1 (3.7 g) was dissolved in THF (20 mL), CDI (2.98 g, ) and DIEA (5.34 g) were added successively, and the reaction was carried out at 50 °C for 1.5 h. The reaction solution was dropped dropwise into ammonia water (40 mL) and stirred for 5 min to complete the reaction. Diluted with water, extracted three times with EA, the organic phases were combined and dried _over anhydrous _Na2SO4 , filtered, and spin-dried. Silica gel column separation and purification (PE:EA=2:1) gave yellow solid M2-2 (2.32 g, 62.93% yield).

ESI-MS m/z: 266.93 [M+H] ⁺ .

Step 3: Synthesis of Compound M2-3

At room temperature, compound 1-1 (4.17 g) was dissolved in THF (10 mL), M2-2 (2.3 g) was added, and the reaction was carried out at 40° C. for 4 h. It was cooled to room temperature, quenched by adding water, extracted three times with EA, and the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and spin-dried. A brown solid M2-3 (3.4 g, crude) was obtained.

ESI-MS m/z: 392.01 [M+H] ⁺ .

Step 4: Synthesis of Compound M2-4

At room temperature, sodium methoxide (2.34 g) was added to a solution of compound M2-3 (3.4 g) in toluene (50 mL), and the mixture was moved to 110° C. and stirred under reflux for 5 hours. Dropped to room temperature, diluted with water, adjusted pH to 6 with 3N HCl, extracted three times with EA, combined organic phases and dried over anhydrous Na ₂ SO ₄ , filtered, and spin-dried to give brown solid M2-4 (2.63 g, crude).

ESI-MS m/z: 374.01 [M+H] ⁺ .

Step 5: Synthesis of Compound M2-5

Under nitrogen protection, DIEA (5 mL) was added to POCl ₃ (50 mL) of compound M2-4 (2.6 g) at room temperature, moved to 110° C. and stirred for 2 hours. After the reaction was completed, the POCl ₃ was directly removed by concentration to obtain a tan solid M2-5 (2.8 g, crude product).

ESI-MS m/z: 391.97 [M+H] ⁺ .

Step 6: Synthesis of Compound M2

DIEA (4.60 g) was added to compound M2-5 (2.8 g), a dioxygen solution of tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (1.93 g) at room temperature Hexacyclic (30 mL) was reacted at room temperature for 5 min. It was quenched by adding water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, spin-dried, and purified by silica gel column separation (DCM:MeOH=10:1) to obtain M2 (3.4 g, 81.9%) as a brown solid Yield).

ESI-MS m/z: 582.16 [M+H] ⁺ .

Example 1: Compound 1 (1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(1 methylpiperidin-4-yl)-8-(2, 2,2-Trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one )Synthesis

Step 1: Synthesis of Compound 1-1

At room temperature, under nitrogen protection, 1-methylpiperidine-4-carboxylic acid (500 mg) was dissolved in SOCl ₂ (2 mL), moved to 70° C., and the reaction was stirred for 1 h. Concentrated in vacuo to remove thionyl chloride to give the desired product, compound 1-1 (520 mg, crude) as a white solid.

Step 2: Synthesis of Compounds 1-2

Compound 1-1 (405.27 mg) was added to compound M1 (450 mg) in THF (10 mL) at room temperature, and the mixture was moved to 40°C and stirred for 4 hours. After the reaction was completed, the THF was directly concentrated to obtain the crude target product as a white solid, namely compound 1-2 (600 mg, crude).

ESI-MS m/z: 483.94 [M+H] ⁺ .

Step 3: Synthesis of Compounds 1-3

At room temperature, sodium methoxide (200.87 mg) was added to compound 1-2 (600 mg) in toluene (15 mL), and the mixture was moved to 110° C. and stirred under reflux for 5 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to obtain the crude target product as a white solid, namely compound 1-3 (600 mg, crude).

ESI-MS m/z: 465.96 [M+H] ⁺ .

Step 4: Synthesis of Compounds 1-4

DIEA (0.5 mL) was added to compound 1-3 (600 mg) in POCl ₃ (5 mL) at room temperature under nitrogen protection, moved to 110° C. and stirred for 3 hours. After the reaction was completed, the POCl ₃ was directly concentrated to remove the target product as a tan solid crude product, namely compound 1-4 (600 mg, crude product).

ESI-MS m/z: 483.96 [M+H] ⁺ .

Step 5: Synthesis of Compounds 1-5

DIEA (480.12 mg) was added to compound 1-4 (600 mg), 1,4- tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (280.25 mg) at room temperature In dioxane (10 mL), the mixture was stirred at room temperature for 3 hours. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (DCM:MeOH=10:1) 1) Purification to obtain the desired target product, compound 1-5 (750 mg, 89.81% yield) as a yellow solid.

ESI-MS m/z: 674.10 [M+H] ⁺ .

Step 6: Synthesis of Compounds 1-6

Under nitrogen protection, trifluoroethanol (244.76 mg) was added to compound 1-5 (550 mg), Cs ₂ CO ₃ (531.45 mg, ) in 1,4-dioxane (5 mL), and moved to 100° C. The reaction was carried out for 30 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo to give the desired target product, compound 1-6 as a yellow solid (590 mg, crude).

ESI-MS m/z: 755.30 [M+H] ⁺ .

Step 7: Synthesis of Compounds 1-7

Under nitrogen protection, Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (63.82 mg) was added to compound 1-6 (590 mg), K ₂ CO ₃ (216.17 mg), 4,4,5,5-tetramethyl In a mixed solution of yl-2-vinyl-1,3,2-dioxaborolane (120.45 mg) in 1,4-dioxane (5.0 mL) and H ₂ O (0.5 mL), Stir at 70°C for 1 h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH = 10:1) purification gave the desired product, compound 1-7 (329 mg, 64.27% yield) as a yellow solid.

ESI-MS m/z: 655.63 [M+H] ⁺ .

Step 8: Synthesis of Compounds 1-8

Under nitrogen protection, Pd(PPh ₃ ) ₄ (116.16 mg) was added to compound 1-7 (329 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 , 5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (344.04 mg), K ₃ PO ₄ (213.38 mg, ) in 1,4-dioxane (3 mL ), H ₂ O (0.75 mL) solution, moved to 85° C. to react for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH = 10:1 ) was purified to give the desired product, compound 1-8 (244 mg, 61.45% yield) as a yellow solid.

ESI-MS m/z: 874.10 [M+H] ⁺ .

Step 9: Synthesis of Compounds 1-9

Compound 1-8 (244 mg) was dissolved in a mixed solvent of DCM (4 mL) and TFA (2 mL) at room temperature, moved to 40° C. to react for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the target product as a yellow solid crude product, namely compound 1-9 (190 mg, crude product).

ESI-MS m/z: 606.38 [M+H] ⁺ .

Step 10: Synthesis of Compound 1

Under nitrogen protection, in an ice-water bath, a solution of acryloyl chloride (17.03 mg) in THF was added to a solution of compound 1-9 (95 mg), THF (4 mL), and saturated Na ₂ CO ₃ (1 mL) solution, and the reaction was completed after the addition. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM/MeOH=10:1) Purification gave the desired product, compound 1 (24.1 mg, 22.96% yield) as a yellow solid.

ESI-MS m/z: 660.37 [M+H]+.

¹ H NMR(500MHz, Methanol-d ₄ )δ8.08(s,1H),7.52(d,J=8.6Hz,1H),7.43-7.32(m,2H),6.40(d,J=17.0,10.3 Hz, 1H), 6.32–6.17 (m, 2H), 5.78–5.69 (m, 2H), 5.09 (d, J=11.1Hz, 1H), 4.86–4.75 (m, 1H), 4.51 (m, 1H) ,4.15(s,2H),3.90(s,2H),3.72(td,J＝6.4,5.2,3.0Hz,4H),3.02(d,J＝11.4Hz,2H),2.85(t,J＝11.5 Hz, 1H), 2.34(s, 3H), 2.25(m, 2H), 2.13(m, 5H), 2.06(m, 3H), 1.90–1.83(m, 4H).

Example 1A: Resolution and Isomer Structure Analysis of Compound 1

Use chromatographic conditions: CHIRALCEL OD (2.5cm I.D.×25cm L, 10μm), UV 254nm, mobile phase Hexane/IPA/DEA=70/30/0.1 (V/V/V), flow rate 20ml/min, resolve 92mg compound 1, to obtain Isomer 1a (56.3 mg) and Isomer 1b (34.3 mg).

Isomer 1a: ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 13.00 (s, 1H), 8.00 (s, 1H), 7.49 (d, J=8.5 Hz, 1H), 7.40 (s, 1H) ,7.32(d,J＝8.6Hz,1H),6.35(dd,J＝17.0,10.3Hz,1H),6.18-6.06(m,2H),5.79-5.66(m,2H),5.11(d,J =11.1Hz,1H),4.97(dq,J=12.3,9.1Hz,1H),4.68(dq,J=12.3,9.1Hz,1H),4.05(s,2H),3.81–3.72(m,6H) ,2.86(dt,J＝11.5,3.1Hz,2H),2.68(tt,J＝11.4,3.6Hz,1H),2.19(s,3H),2.04(s,3H),1.98(dt,J＝6.7 , 4.1 Hz, 8H), 1.85 (qd, J=13.0, 3.9 Hz, 2H). ESI-MS m/z: 660.4 [M+H]+. Chromatographic conditions: CHIRALCEL OD (2.5cm ID×25cm L, 10μm), UV 254nm, mobile phase Hexane/IPA/DEA=70/30/0.1 (V/V/V), flow rate 20ml/min, retention time 6.39min ( back peak);

Isomer 1b: ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 13.01 (s, 1H), 8.00 (s, 1H), 7.50 (d, J=8.5 Hz, 1H), 7.41 (s, 1H) ,7.32(d,J＝8.6Hz,1H),6.35(dd,J＝17.0,10.3Hz,1H),6.19–6.06(m,2H),5.80–5.60(m,2H),5.11(d,J =11.2Hz,1H),4.98(dq,J=12.2,9.1Hz,1H),4.69(dq,J=12.2,9.2Hz,1H),4.05(s,2H),3.76(d,J=7.1Hz ,6H),2.85(dd,J＝8.9,5.8Hz,2H),2.68(tt,J＝11.6,3.7Hz,1H),2.19(s,3H),2.05(s,3H),2.02–1.96( m, 8H), 1.85 (qd, J=13.2, 4.0 Hz, 2H). ESI-MS m/z: 660.4 [M+H]+. Chromatographic conditions: CHIRALCEL OD (2.5cm ID×25cm L, 10μm), UV 254nm, mobile phase Hexane/IPA/DEA=70/30/0.1 (V/V/V), flow rate 20ml/min, retention time 4.09min ( Peak first);

A single crystal of the DMSO solvate of the isomer 1a was obtained by culturing the single crystal by the liquid surface diffusion method in the DMSO/H ₂ O system. The crystal size for the crystal diffraction experiment was 0.03 × 0.19 × 0.23 mm, and the single crystal was tested with a Bruker D8 Venture Photon II diffractometer. The light source was CuK _α radiation, and the scanning method was

scanning. The crystal structure was analyzed by the direct method (Shelxs97), and all 100 non-hydrogen atomic positions were obtained. The crystal belongs to the monoclinic system, the space group is P2 ₁ , and the unit cell parameters are: a=18.4614(13), b=11.8865(8),

α=γ=90°, β=110.112(3)°, unit cell volume

The number of asymmetric units in the unit cell is Z=2. The least squares method was used to correct the structural parameters and identify the atomic species, and the geometric calculation method and the difference Fourier method were used to obtain all the hydrogen atom positions. The results of single crystal structure analysis show that the molecular arrangement in the crystalline state belongs to the first space group, the compound should have optical activity, and the Flack coefficient is 0.044(12), which can confirm that the absolute configuration of the compound in the crystal is the S configuration.

Example 2: Compound 2 (1-(7-(2-cyclohexyl-7-(5-methyl-1H-indazol-4-yl)-8-(2,2,2-trifluoroethoxy) )-6-vinylquinazolin-4-yl)-2,-1,7-diazaspiro[3.5]nonan-2-yl)prop-2-en-1-one) synthesis

Step 1: Synthesis of Compound 2-1

At room temperature, cyclohexanecarbonyl chloride (367.63 mg) was added to compound M1 (300 mg) in THF (10 mL), and the mixture was moved to 40°C and stirred for 4 hours. After the reaction was completed, the THF was directly concentrated to obtain the crude target product as a white solid, namely compound 2-1 (390 mg, crude).

ESI-MS m/z: 469.00 [M+H] ⁺ .

Step 2: Synthesis of Compound 2-2

At room temperature, sodium methoxide (224.56 mg) was added to 2-1 (390 mg) in toluene (15 mL), and the mixture was moved to 110° C. and stirred under reflux for 5 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to obtain the crude target product as a white solid, namely compound 2-2 (350 mg, crude).

ESI-MS m/z: 450.94 [M+H] ⁺ .

Step 3: Synthesis of Compounds 2-3

DIEA (1.0 mL) was added to compound 2-2 (350 mg) in POCl ₃ (10.0 mL) at room temperature under nitrogen protection, moved to 110° C. and stirred for 12 hours. After the reaction was completed, the POCl ₃ was directly concentrated to remove the target product as a tan solid crude product, namely compound 2-3 (340 mg, crude product).

ESI-MS m/z: 468.87 [M+H]+.

Step 4: Synthesis of Compounds 2-4

DIEA (279 mg) was added to compound 2-3 (340 mg), 1,4-dioxo tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (163 mg) at room temperature Hexacyclic (10 mL) was stirred at room temperature for 5 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (PE:EA=10:1-15:1) Purification gave the desired target product, compound 2-4 (348 mg, 42.62% yield) as a yellow solid.

ESI-MS m/z: 659.06 [M+H] ⁺ .

Step 5: Synthesis of Compounds 2-5

Under nitrogen protection, trifluoroethanol (158.4 mg) was added to compound 2-4 (348 mg), Cs ₂ CO ₃ (343.92 mg, ) in 1,4-dioxane (5 mL), and moved to 100° C. The reaction was carried out for 30 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo to give the desired target product, compound 2-5 (400 mg, crude) as a yellow solid.

ESI-MS m/z: 739.01 [M+H] ⁺ .

Step 6: Synthesis of Compounds 2-6

Under nitrogen protection, Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (63.82 mg) was added to compound 2-5 (400 mg), K ₂ CO ₃ (149.53 mg), 4,4,5,5-tetramethyl In a mixed solution of yl-2-vinyl-1,3,2-dioxaborolane (83.32 mg, ) in 1,4-dioxane (4.0 mL), H ₂ O (0.4 mL) , and stirred at 70 °C for 1 h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, and the residue was subjected to flash silica gel column chromatography ( PE:EA=10:1-15:1) was purified to give the desired target product, compound 2-6 (213 mg, 61.56% yield) as a yellow solid.

ESI-MS m/z: 639.01 [M+H] ⁺ .

Step 7: Synthesis of Compounds 2-7

Under nitrogen protection, Pd(PPh ₃ ) ₄ (116.16 mg) was added to compound 2-6 (213 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 ,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (227.96 mg), K ₃ PO ₄ (141.39 mg, ) in 1,4-dioxane (2 mL ), H ₂ O (0.5 mL) solution, moved to 85° C. to react for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, the concentrate was passed through pre-TLC (PE:EA=2:1 ) was purified to give the desired product, compound 2-7 (180 mg, 69.74% yield) as a yellow solid. ESI-MS m/z: 775.42 [M+H] ⁺ .

Step 8: Synthesis of Compounds 2-8

Compound 2-7 (180 mg) was dissolved in a mixed solvent of DCM (6 mL) and TFA (3 mL) at room temperature, moved to 40° C. to react for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the target product as a yellow solid crude product, namely compound 2-8 (135 mg, crude product).

ESI-MS m/z: 591.40 [M+H] ⁺ .

Step 9: Synthesis of Compounds 2-9

Under nitrogen protection, in an ice-water bath, a solution of acryloyl chloride (17.03 mg) in THF was added to a solution of compound 2-8 (135 mg, ), THF (4 mL), and saturated Na ₂ CO ₃ (1 mL) solution, and the reaction was completed after the addition. . The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM/MeOH=10:1) Purification gave the desired product, compound 2 (66.71 mg, 44.36% yield) as a yellow solid. ESI-MS m/z: 645.44 [M+H] ⁺ . 1HNMR(500MHz, CDCl3)δ: 7.94(s,1H), 7.45-7.48(m,2H), 7.36-7.36(m,1H), 6.37-6.41(m,1H), 6.22-6.28(m,2H) ,5.70-5.73(m,1H),5.62-5.66(m,1H),5.05-5.07(m,1H),4.83-4.91(m,1H),4.47-4.54(m,1H),4.04(s, 2H), 3.94(s, 2H), 3.70-3.79(m, 4H), 2.81-2.87(m, 1H), 2.15(s, 3H), 2.07-2.09(m, 2H), 1.85-1.87(m, 2H), 1.74-1.77 (m, 1H), 1.61-1.71 (m, 3H), 1.41-1.48 (m, 3H), 1.25-1.37 (m, 3H).

Example 3: Compound 3 (1-(7-(2-tetrahydrofuran-3-yl-7-(5-methyl-1H-indazol-4-yl)-8-(2,2,2-trifluoro Ethoxy)-6-vinylquinazolin-4-yl)-2,-1,7-diazaspiro[3.5]nonan-2-yl)prop-2-en-1-one) of synthesis

Step 1: Synthesis of Compound 3-1

At room temperature, tetrahydrofuran-3-carbonyl chloride (402 mg) was added to compound M1 (300 mg) in THF (10 mL), and the mixture was moved to 40°C and stirred for 4 hours. After the reaction was completed, the THF was directly concentrated to obtain the crude target product as a yellow solid, namely compound 3-1 (460 mg, crude).

ESI-MS m/z: 456.94 [M+H] ⁺ .

Step 2: Synthesis of Compound 3-2

At room temperature, sodium methoxide (224.56 mg) was added to compound 3-1 (460 mg, 0.83 mmol) in toluene (10 mL), and the mixture was moved to 110° C. and stirred under reflux for 8 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to obtain the crude target product as a white solid, namely compound 3-2 (200 mg, crude).

ESI-MS m/z: 438.96 [M+H]+.

Step 3: Synthesis of Compound 3-3

DIEA (1.0 mL) was added to compound 3-2 (200 mg) in POCl ₃ (10.0 mL) at room temperature under nitrogen protection, moved to 110° C. and stirred for 12 hours. After the reaction was completed, the POCl ₃ was directly concentrated to remove the target product as a tan solid crude product, namely compound 3-3 (230 mg, crude product).

ESI-MS m/z: 456.87 [M+H] ⁺ .

Step 4: Synthesis of Compounds 3-4

DIEA (279 mg) was added to compound 3-3 (200 mg), 1,4-dioxo tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (90 mg) at room temperature Hexacyclic (2 mL) was stirred at room temperature for 5 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo , the residue was purified by climbing plate to give the desired target product as a yellow solid, compound 3 -4 (88 mg, 34.1% yield).

ESI-MS m/z: 647.14 [M+H] ⁺ .

Step 5: Synthesis of Compounds 3-5

Under nitrogen protection, trifluoroethanol (68 mg) was added to compound 3-4 (88 mg), Cs ₂ CO ₃ (88 mg) in 1,4-dioxane (2 mL), moved to 100° C. to react for 30 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo to give the desired target product, compound 3-5 (97 mg, crude) as a yellow solid.

ESI-MS m/z: 727.08 [M+H] ⁺ .

Step 6: Synthesis of Compounds 3-6

Under nitrogen protection, Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (10 mg) was added to compound 3-5 (90 mg), K ₂ CO ₃ (34 mg, ), 4,4,5,5-tetramethyl - A mixed solution of 2-vinyl-1,3,2-dioxaborolane (19 mg) in 1,4-dioxane (2.5 mL), H ₂ O (0.5 mL) at 70 °C and stirred for 1 h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, and the residue was subjected to flash silica gel column chromatography ( PE:EA=10:1-3:1) was purified to give the desired target product, compound 3-6 (213 mg, 64.4% yield) as a yellow solid.

ESI-MS m/z: 627.21 [M+H] ⁺ .

Step 7: Synthesis of Compounds 3-7

Under nitrogen protection, Pd(PPh ₃ ) ₄ (18.5 mg) was added to compound 3-6 (50 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 , 5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (54.6 mg), 1,4-dioxane (2 mL) in _K3PO4 ( ₃₄ mg), H ₂ O (0.5 mL) solution, moved to 85° C. to react for about 4 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, the concentrate was passed through pre-TLC (PE:EA=2:1 ) was purified to give the desired product, compound 3-7 (45 mg, 73.8% yield) as a yellow solid.

ESI-MS m/z: 763.39 [M+H] ⁺ .

Step 8: Synthesis of Compounds 3-8

Compound 3-7 (45 mg) was dissolved in a mixed solvent of DCM (1 mL) and TFA (0.5 mL) at room temperature, moved to 40° C. to react for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the target product as a yellow solid crude product, namely compound 3-8 (56 mg, crude product). ESI-MS m/z: 579.32 [M+H] ⁺ .

Step 9: Synthesis of Compounds 3-9

Under nitrogen protection, in an ice-water bath, a solution of acryloyl chloride (5.85 mg) in THF was added to a solution of compound 3-8 (56 mg, ), THF (2 mL), and saturated Na ₂ CO ₃ (0.5 mL) solution, and the reaction was performed after the addition. completely. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM/MeOH=10:1) Purification gave the desired product, compound 3 (12.05 mg, 32.2% yield) as a yellow solid. ESI-MS m/z: 633.41 [M+H]+.

1H NMR(500MHz, Chloroform-d)δ7.88(s,1H),7.40(d,J=7.5Hz,2H),7.19(s,1H),6.32(dd,J=17.0,1.9Hz,1H) ,6.18(m,J＝17.0,10.6,4.9Hz,2H),5.65(dd,J＝10.3,1.9Hz,1H),5.58(dd,J＝17.4,1.0Hz,1H),5.01(dd,J =10.9,1.0Hz,1H),4.78–4.64(m,1H),4.34(m,1H),4.20(td,J=8.1,3.7Hz,1H),4.05(dd,J=8.3,6.8Hz, 1H), 4.02–3.95 (m, 3H), 3.94–3.89 (m, 1H), 3.87 (s, 2H), 3.75 (td, J=10.0, 9.0, 5.5Hz, 2H), 3.71–3.62 (m, 3H), 2.41 (m, 1H), 2.29 (m, 1H), 2.08 (s, 3H), 1.18 (d, J=7.2Hz, 4H).

Example 4: Compound 4 (1-(7-(6-ethyl-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl) -8-(2,2,2-Trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-ene-1- ketone) synthesis

Step 1: Synthesis of Compound 4-1

Pd/C (80 mg) was added to a solution of compound 1-9 (95 mg) in MeOH (10 mL) at room temperature under a hydrogen atmosphere, and stirred at room temperature for 2 hours. After the reaction was completed, the Pd/C was removed by filtration, and the Pd/C was washed with MeOH, and the filtrate was concentrated to obtain a yellow solid crude product, namely compound 4-1 (70 mg, crude product).

ESI-MS m/z: 608.43 [M+H] ⁺ .

Step 2: Synthesis of Compound 4

Under nitrogen protection, in an ice-water bath, a THF solution of acryloyl chloride (12.51 mg) was added to compound 4-1 (70 mg), THF (4 mL), saturated Na ₂ CO ₃ (1 mL) solution, and the reaction was completed after the addition. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM/MeOH=10:1) Purification gave the desired product, compound 4 (18.3 mg, 23.79% yield) as a yellow solid.

ESI-MS m/z: 662.37 [M+H] ⁺ .

Example 5: Compound 5 (1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(tetrahydro-2H-pyran-4-yl)-8-( 2,2,2-Trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-ene-1 -ketone) synthesis

Step 1: Synthesis of Compound 5-1

At room temperature, tetrahydropyran-4-carbonyl chloride (248.38 mg, 1.67 mmol) was added to compound M1 (300 mg, ) in THF (3 mL), moved to 40°C and stirred for 4 hours. After the reaction was completed, the THF was directly concentrated to obtain the crude target product as a white solid, namely compound 5-1 (390 mg, crude).

ESI-MS m/z: 470.92 [M+H] ⁺ .

Step 2: Synthesis of Compound 5-2

At room temperature, sodium methoxide (224.56 mg) was added to a solution of compound 5-1 (390 mg) in toluene (4 mL), and the mixture was moved to 110° C. and stirred under reflux for 5 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to obtain the crude target product as a white solid, namely compound 5-2 (350 mg, crude).

ESI-MS m/z: 452.96 [M+H] ⁺ .

Step 3: Synthesis of Compound 5-3

DIEA (0.4 mL) was added to compound 5-2 (350 mg, 0.77 mmol) in POCl ₃ (4 mL) at room temperature under nitrogen protection, moved to 110° C. and stirred for 12 hours. After the reaction was completed, the POCl ₃ was directly concentrated to remove the target product as a tan solid crude product, namely compound 5-3 (210 mg, crude product).

ESI-MS m/z: 470.95 [M+H] ⁺ .

Step 4: Synthesis of Compounds 5-4

DIEA (115.13 mg) was added to compound 5-3 (210 mg), 1,4- tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (120.96 mg) at room temperature In dioxane (3 mL), the mixture was stirred at room temperature for 30 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (PE:EA=10:1-15:1) Purification gave the desired target product, compound 5-4 (110 mg, 37.34% yield) as a yellow solid.

ESI-MS m/z: 661.17 [M+H] ⁺ .

Step 5: Synthesis of Compounds 5-5

Under nitrogen protection, trifluoroethanol (49.92 mg) was added to compound 5-4 (110 mg), Cs ₂ CO ₃ (108.39 mg, ) in 1,4-dioxane (2 mL), and moved to 100° C. The reaction was carried out for 30 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo to give the desired target product, compound 5-5 (105 mg, crude) as a yellow solid.

ESI-MS m/z: 741.14 [M+H] ⁺ .

Step 6: Synthesis of Compounds 5-6

Under nitrogen protection, Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (11.56 mg) was added to compound 5-5 (105 mg), K ₂ CO ₃ (39.15 mg), 4,4,5,5-tetramethyl In a mixed solution of yl-2-vinyl-1,3,2-dioxaborolane (21.81 mg) in 1,4-dioxane (2 mL) and H ₂ O (0.4 mL), Stir at 70°C for 1 h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, and the residue was subjected to flash silica gel column chromatography ( PE:EA=10:1-15:1) was purified to give the desired target product, compound 5-6 (60 mg, 66.04% yield) as a yellow solid. ESI-MS m/z: 641.25 [M+H] ⁺ .

Step 7: Synthesis of Compounds 5-7

Under nitrogen protection, Pd(PPh ₃ ) ₄ (21.60 mg) was added to compound 5-6 (60 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 ,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (64.02 mg), K ₃ PO ₄ (39.71 mg, ) in 1,4-dioxane (2 mL ), H ₂ O (0.2 mL) solution, moved to 85°C for reaction for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, the concentrate was passed through pre-TLC (PE:EA=2:1 ) was purified to give the desired product, compound 5-7 (25 mg, 34.41% yield) as a yellow solid.

ESI-MS m/z: 777.42 [M+H] ⁺ .

Step 8: Synthesis of Compounds 5-8

Compound 5-7 (25 mg) was dissolved in a mixed solvent of DCM (1 mL) and TFA (0.5 mL) at room temperature, moved to 40° C. to react for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the target product as a yellow oily crude product, namely compound 5-8 (20 mg, crude product).

ESI-MS m/z: 593.33 [M+H] ⁺ .

Step 9: Synthesis of Compound 5

Under nitrogen protection, in an ice-water bath, a solution of acryloyl chloride (3.20 mg) in THF was added to a solution of compound 5-8 (20 mg, ), THF (2 mL), and saturated Na ₂ CO ₃ (0.5 mL) solution, and the reaction was performed after the addition. completely. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM/MeOH=10:1) Purification gave the desired product, compound 5 (5.2 mg, 33.48% yield) as a yellow solid.

ESI-MS m/z: 647.33 [M+H] ⁺ .

Example 6: Compound 6(1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(N-methylpyrrol-3-yl)-8-(2, 2,2-Trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one )Synthesis

Step 1: Synthesis of Compound 6-1

At room temperature under nitrogen protection, N-methylpyrrole-3-carboxylic acid (600 mg) was dissolved in SOCl ₂ (5.5 mL), the temperature was raised to 70° C., and the reaction was stirred for 1 h. Concentrated in vacuo to remove thionyl chloride to give the desired product as a white solid, compound 6-1 (68 mg, crude).

Step 2: Synthesis of Compound 6-2

Compound 6-1 (369 mg) was added to compound M1 (300 mg) in THF (5 mL) at room temperature and stirred for 16 hours. After the reaction was completed, the THF was removed by rotary evaporation. Using dichloromethane:methanol=90:10 as a developing solvent, column chromatography was used to obtain a pale yellow solid target product, namely compound 6-2 (262 mg).

ESI-MS m/z: 469.93, 471.94 [M+H] ⁺ .

Step 3: Synthesis of Compound 6-3

At room temperature, sodium methoxide (151 mg) was added to a solution of compound 6-2 (262 mg) in toluene (10 mL), the temperature was raised to 110° C. and refluxed and stirred for 16 hours. After the reaction was completed, the mixture was poured into 40 mL of ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to obtain the target product as a pale yellow solid, namely compound 6-3 (240 mg, crude product) .

ESI-MS m/z: 451.96, 453.91 [M+H] ⁺ .

Step 4: Synthesis of Compounds 6-4

Under nitrogen protection at room temperature, DIEA (0.5 mL) was added to a solution of compound 6-3 (226 mg) in POCl ₃ (5 mL), and the temperature was raised to 110° C. and stirred for 3 hours. After the reaction was completed, the POCl ₃ was removed by rotary evaporation, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was collected, washed with saturated brine, dried over Na ₂ SO ₄ and concentrated in vacuo to obtain the target product as a brown solid , namely compound 6-4 (116 mg, crude).

ESI-MS m/z: 469.96, 471.94 [M+H] ⁺ .

Step 5: Synthesis of Compounds 6-5

DIEA (96 mg) was added to compound 6-4 (116 mg), tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (56 mg) in dioxane (5 mL) at room temperature ) solution and stirred at room temperature for 2 hours. After the reaction, dioxane was spin-dried, ice water was added, the mixture was extracted with ethyl acetate/methanol=10:1, and the organic phase was collected and spin-dried. The residue was purified by silica gel column chromatography (developing solvent: DCM:MeOH=10:1) to obtain the yellow solid target product, namely compound 6-5 (58 mg).

ESI-MS m/z: 660.08, 662.07 [M+H] ⁺ .

Step 6: Synthesis of Compounds 6-6

Compound 6-5 (56 mg) was added to the well-dried reaction flask, dissolved in dioxane (3 mL), then trifluoroethanol (42 mg) and Cs ₂ CO ₃ (55 mg) were added, and the temperature was raised to 100° C. to react for 1 h. After completion of the reaction, the reaction solution was diluted with THF, and Cs ₂ CO ₃ was removed by filtration. After the filtrate was spin-dried, the sample was dried using a diaphragm pump to obtain the yellow solid target product, namely compound 6-6 (60 mg).

ESI-MS m/z: 740.03, 742.07 [M+H] ⁺ .

Step 7: Synthesis of Compounds 6-7

Under _N2 protection, into a dry reaction flask was added compound 6-6 (60 mg), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (13 mg), dissolved in dioxane (5 mL), then K ₂ CO ₃ (22.40 mg), H ₂ O (0.5 mL) were added, and after the dispersion was uniform, 1,1'-bis-diphenylphosphinodioxin was added Iron palladium chloride (5.93 mg), N ₂ displacement system three times. The temperature was raised to 70 °C and stirred for 1 h. After the reaction was completed, the reaction solution was cooled to room temperature, added with water, extracted with ethyl acetate:methanol=10:1, back-extracted with saturated brine, collected the organic phase, dried over Na ₂ SO ₄ , filtered and spin-dried, and the concentrate was Purification by pre-TLC (developing solvent: DCM/MeOH=10:1) gave the product as a yellow solid, namely compound 6-7 (44 mg).

ESI-MS m/z: 640.11, 642.13 [M+H] ⁺ .

Step 8: Synthesis of Compounds 6-8

Compound 6-7 (44 mg) was added to a nitrogen-protected reaction flask, dissolved in dioxane (5 mL), followed by 5-methyl-1-tetrahydropyran-2-yl-4-(4, 4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (47 mg), K ₃ PO ₄ (29 mg), deionized water (0.5 mL) and Pd ( _PPh3 ) ₄ (8 mg). The temperature was raised to 85°C for 3 hours. The reaction mixture was cooled to room temperature, extracted with ethyl acetate:methanol = 10:1 after adding water, back-extracted with saturated brine, the organic phase was collected, dried over Na ₂ SO ₄ , filtered and spun dry, the concentrate was passed through pre-TLC (developing solvent is DCM/MeOH=10:1) and purified to obtain a yellow solid product, namely compound 6-8 (18 mg). ESI-MS m/z: 776.87 [M+H] ⁺ .

Step 9: Synthesis of Compounds 6-9

Compound 6-8 (18 mg) was dissolved in a mixed solvent of DCM (2 mL) and TFA (1 mL) at room temperature, and the temperature was raised to 40° C. to react for 1 h. After the reaction was completed, the solvent was concentrated and removed to obtain a yellow solid crude product, that is, compound 6-9 (21 mg, crude product) was directly used in the next feeding. ESI-MS m/z: 592.94 [M+H] ⁺ .

Step 10: Synthesis of Compound 6

Under nitrogen protection and ice-water bath, a solution of acryloyl chloride (2.0 mg) in THF (1 mL) was added dropwise to a mixed solution of compound 6-9 (13 mg,) in THF (4 mL) and saturated NaHCO ₃ (1 mL), and stirred for 10 minute. The reaction solution was poured into water, extracted with ethyl acetate:methanol=10:1, back-extracted with saturated brine, the organic phase was collected, dried over Na ₂ SO ₄ , filtered, and spin-dried, and the concentrate was passed through pre-TLC (developing solvent). Purification with DCM/MeOH=10:1) gave the product as a pale yellow solid, compound 6 (7 mg, LC-MS purity 96.2%).

ESI-MS m/z: 646.34 [M+H] ⁺ .

Example 7: Compound 7 (1-(7-(2-(1-ethylpiperidin-4-yl)-7-(5-methyl-1H-indazol-4-yl)-8-(2 ,2,2-Trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-ene-1- ketone) synthesis

Step 1: Synthesis of Compound 7-1

At room temperature, iodoethane (3.27 g), K ₂ CO ₃ (2.89 g) was added to a solution of methyl piperidine-4-carboxylate (1.5 g, ) in EtOH (15 mL), moved to 85 ℃ reflux reaction for 1h. Concentration in vacuo to remove EtOH afforded the desired product 7-1 as a white solid (1.7 g, crude).

ESI-MS m/z: 172.06 [M+H] ⁺ .

Step 2: Synthesis of Compound 7-2

LiOH (447.57 mg) was added to a solution of compound 7-1 (1.7 g) in MeOH (10 mL) at room temperature, and the mixture was moved to 50° C. to react for 1 hour. Diluted hydrochloric acid solution equivalent to LiOH was added to it to free the target product, directly spin-dried, and the residue was purified by flash silica gel column chromatography (DCM:MeOH=92:8-90:10) to obtain the desired target as a yellow solid Product 7-2 (700 mg, 44.85% yield).

ESI-MS m/z: 158.15 [M+H] ⁺ .

Step 3: Synthesis of Compound 7-3

At room temperature, under nitrogen protection, compound 7-2 (400 mg) was dissolved in SOCl ₂ (3 mL), moved to 70° C., and the reaction was stirred for 1 h. Concentration in vacuo to remove thionyl chloride gave the desired product 7-3 as a white solid (450 mg, crude).

Step 4: Synthesis of Compounds 7-4

Compound 7-3 (293.63 mg) was added to compound M1 (300 mg) in THF (10 mL) at room temperature, and the mixture was moved to 45°C and stirred for 12 hours. A large amount of solid was precipitated in the reaction, which was filtered with suction, and the solid was the target product 7-4 (150 mg, crude product).

ESI-MS m/z: 498.00 [M+H] ⁺ .

Step 5: Synthesis of Compounds 7-5

At room temperature, sodium methoxide (81.34 mg) was added to toluene (5 mL) of compound 7-4 (150 mg), and the mixture was moved to 110° C. and stirred under reflux for 4 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to give the crude target product 7-5 as a brown solid (150 mg, crude).

ESI-MS m/z: 480.00 [M+H] ⁺ .

Step 6: Synthesis of Compounds 7-6

DIEA (1.0 mL) was added to compound 7-5 (150 mg) in POCl ₃ (5 mL) at room temperature under nitrogen protection, moved to 110° C. and stirred for 3 hours. After the reaction was completed, it was directly concentrated to remove POCl ₃ , diluted with EA, extracted with water three times, combined with the organic phases, dried and concentrated to obtain the target product 7-6 (150 mg, crude product) as a white solid.

ESI-MS m/z: 497.99 [M+H] ⁺ .

Step 7: Synthesis of Compounds 7-7

DIEA (116.65 mg) was added to compound 7-6 (150 mg), tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (68.09 mg) in dioxane at room temperature (5 mL), moved to 45°C and stirred for 1 hour. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (DCM:MeOH=10:1) 1) Purification gave the desired target product 7-7 as a yellow solid (50 mg, 24.14% yield).

ESI-MS m/z: 688.14 [M+H] ⁺ .

Step 8: Synthesis of Compounds 7-8

Under nitrogen protection, trifluoroethanol (21.80 mg) was added to compound 7-7 (50 mg), Cs ₂ CO ₃ (47.33 mg) in dioxane (1 mL), moved to 100° C. to react for 3 h. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo to give the desired target product 7-7 as a yellow solid (55 mg, Crude).

ESI-MS m/z: 768.18 [M+H] ⁺ .

Step 9: Synthesis of Compounds 7-9

Under nitrogen protection, Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (5.84 mg) was added to compound 7-8 (55 mg), K ₂ CO ₃ (19.78 mg), 4,4,5,5-tetramethyl A mixed solution of yl-2-vinyl-1,3,2-dioxaborolane (11.02 mg) in dioxane (1.0 mL) and H ₂ O (0.1 mL) was stirred at 70°C 1h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH = 10:1) purification gave the desired product 7-9 as a yellow solid (25 mg, 37.39 μmol, 52.24% yield).

ESI-MS m/z: 668.18 [M+H] ⁺ .

Step 10: Synthesis of Compounds 7-10

Under nitrogen protection, Pd(PPh ₃ ) ₄ (8.64 mg) was added to compound 7-9 (25 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 ,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (25.59 mg), K ₃ PO ₄ (15.87 mg, ) in dioxane (1 mL), H ₂ O (0.25 mL) solution, moved to 85°C to react for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH=10:1 ) was purified to give the desired product 7-10 (20 mg, 66.53% yield) as a yellow solid. ESI-MS

m/z: 804.56[M+H] ⁺ .

Step 11: Synthesis of Compounds 7-11

Compound 7-10 (20 mg) was dissolved in a mixed solvent of DCM (1 mL) and TFA (0.5 mL) at room temperature, and moved to 40° C. to react for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the solvent to obtain the target product 7-11 (18 mg, crude product) as a yellow solid.

ESI-MS m/z: 620.28 [M+H] ⁺ .

Step 12: Synthesis of Compound 7

Under nitrogen protection, in an ice-water bath, a solution of acryloyl chloride (3.15 mg) in THF was added to a solution of compound 9-11 (18 mg, ), THF (2 mL), and saturated Na ₂ CO ₃ (0.5 mL) solution, and the reaction was performed after the addition. completely. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM/MeOH=10:1) Purification gave the desired product 7 as a yellow solid (10.7 mg, 52.69% yield).

ESI-MS m/z: 674.37 [M+H] ⁺ .

¹ H NMR (500MHz, DMSO-d ₆ ) δ 13.05(s, 1H), 8.30(s, 1H), 8.01(s, 1H), 7.50(d, J=8.5Hz, 1H), 7.40(s, 1H), 7.33 (d, J=8.5Hz, 1H), 6.35 (m, 1H), 6.18–6.05 (m, 2H), 5.82–5.66 (m, 2H), 5.11 (d, J=11.1Hz, 1H) ), 4.97(m, 1H), 4.68(m, 1H), 3.23–3.05(m, 4H), 2.80(m, 1H), 2.25(m, 2H), 2.05(d, J=6.7Hz, 5H) ,1.95(m,6H),1.62–1.55(m,1H),1.32(m,1H),1.07(t,J=7.2Hz,3H),0.94(t,J=7.3Hz,2H).

Example 8: Compound 8 (1-(7-(6-Chloro-8-ethoxy-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidine) Synthesis of -4-yl)-quinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one)

Step 1: Synthesis of Compound 8-1

Under nitrogen protection, ethanol (395 mg) was added to compound M2 (1.0 g), Cs ₂ CO ₃ (1.68 g) in dioxane (10 mL), moved to 80° C. to react for 1 h. It was lowered to room temperature, water was slowly added, a solid was precipitated, filtered, and dried to obtain a yellow solid 8-1 (0.96 g, crude product).

ESI-MS m/z: 608.19 [M+H] ⁺ .

Step 2: Synthesis of Compound 8-2

Under nitrogen protection, Pd(PPh ₃ ) ₄ (341 mg) was added to compound 8-1 (900 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole ( _758.6 mg), _K3PO4 (941.1 mg, ) in dioxane (20 mL), _H2O (5mL) solution, moved to 85°C to react for 4h. It was cooled to room temperature, diluted with water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, spun dry, separated and purified by silica gel column (DCM:MeOH=10:1) to obtain yellow solid 8-2 (665 mg, 49.9% yield).

ESI-MS m/z: 744.39 [M+H] ⁺ .

Step 3: Synthesis of Compound 8-3

At room temperature, compound 8-2 (120 mg) was dissolved in a mixed solvent of DCM (6 mL) and TFA (3 mL), moved to 40° C. and reacted for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the solvent to obtain a brown oil crude product 8-3 (73 mg, crude product).

ESI-MS m/z: 560.28 [M+H] ⁺ .

Step 4: Synthesis of Compound 8

Under nitrogen protection, a solution of acryloyl chloride (12.1 mg) in THF (1 mL) was added to a solution of compound 8-3 (73 mg), THF (4 mL), saturated Na ₂ CO ₃ (1 mL) in an ice-water bath, maintaining the temperature Stir for 5 minutes. It was diluted with water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, spun dry, and separated and purified by pre-TLC (DCM:MeOH=10:1) to obtain 8 (36.5 mg, 43.3 mg) as a white solid. %Yield).

ESI-MS m/z: 614.40 [M+H] ⁺ .

Example 9: Compound 9 (1-(7-(8-ethoxy-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl) )-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one) synthesis

Step 1: Synthesis of Compound 9-1

Under nitrogen protection, Sphos Pd G2 (51.7 mg) was added to compound 8-2 (300 mg), K ₃ PO ₄ (228.4 mg, ), vinylborate (110.49 mg) in dioxane (4 mL) , H ₂ O (1 mL) mixed solution, stirred at 100 ° C for 4 h. It was cooled to room temperature, diluted with water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, spun dry, and separated and purified by pre-TLC (DCM:MeOH=10:1) to obtain a white solid 9- 1 (185 mg, 70.08% yield).

ESI-MS m/z: 736.45 [M+H] ⁺ .

Step 2: Synthesis of Compound 9-2

At room temperature, compound 9-1 (185 mg) was dissolved in a mixed solvent of DCM (4 mL) and TFA (2 mL), moved to 40° C. and reacted for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the solvent to obtain a brown oil crude product 9-2 (138 mg, crude product).

ESI-MS m/z: 552.34 [M+H] ⁺ .

Step 3: Synthesis of Compound 9

Under nitrogen protection, a solution of acryloyl chloride (22.6 mg) in THF (1 mL) was added to a solution of compound 9-2 (138 mg), THF (4 mL), saturated Na ₂ CO ₃ (1 mL) in an ice-water bath, maintaining the temperature Stir for 5 minutes. It was diluted with water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, spin-dried, and separated and purified by pre-TLC (DCM:MeOH=10:1) to obtain a white solid 9 (62.5 mg, 38.2 %Yield).

ESI-MS m/z: 606.45 [M+H] ⁺ .

¹ H NMR (500MHz, Chloroform-d)δ7.90(s,1H),7.51(s,1H),7.46(d,J=8.5Hz,1H),7.37-7.34(m,1H),6.39(dd ,J=17.0,1.9Hz,1H),6.24(m,2H),5.71(dd,J=10.2,1.9Hz,1H),5.62(d,J=17.4Hz,1H),5.03(d,J= 11.0Hz, 1H), 4.22(m, 1H), 4.03(s, 2H), 3.93(s, 2H), 3.91– 3.84(m, 1H), 3.75(m, 4H), 3.00(t, J=6.1 Hz, 2H), 2.87(m, 1H), 2.35(s, 3H), 2.16(s, 3H), 2.12–2.08(m, 5H), 2.03(m, 4H), 1.33(m, 4H).

Example 10: Compound 10 (1-(7-(6-Chloro-8-methoxy-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidine) Synthesis of -4-yl)-quinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one)

Step 1: Synthesis of Compound 10-1

Under nitrogen protection, methanol (275 mg) was added to compound M2 (1.0 g), Cs ₂ CO ₃ (1.68 g) in dioxane (10 mL), and moved to 70° C. to react for 5 h. It was lowered to room temperature, water was slowly added, a solid was precipitated, filtered, and dried to obtain a yellow solid 10-1 (0.91 g, crude product).

ESI-MS m/z: 594.18 [M+H] ⁺ .

Step 2: Synthesis of Compound 10-2

Under nitrogen protection, Pd(PPh ₃ ) ₄ (77.69 mg) was added to compound 10-1 (200 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 ,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (172.5 mg), K ₃ PO ₄ (214.1 mg, ) in dioxane (4 mL), H ₂ O (1 mL) solution, moved to 85°C and reacted for 4h. It was cooled to room temperature, diluted with water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, spun dry, separated and purified on a silica gel column (DCM:MeOH=10:1) to obtain a yellow solid 10-2 (202 mg, 79.8% yield).

ESI-MS m/z: 730.38 [M+H] ⁺ .

Step 3: Synthesis of Compound 12-3

At room temperature, compound 10-2 (200 mg) was dissolved in a mixed solvent of DCM (6 mL) and TFA (3 mL), moved to 40° C. to react for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the solvent to obtain a brown oil crude product 10-3 (149 mg, crude product).

ESI-MS m/z: 546.27 [M+H] ⁺ .

Step 4: Synthesis of Compound 10

Under nitrogen protection, a solution of acryloyl chloride (24.7 mg) in THF (1 mL) was added to a solution of compound 12-3 (149 mg), THF (4 mL), saturated Na ₂ CO ₃ (1 mL) in an ice-water bath, maintaining the temperature Stir for 5 minutes. Diluted with water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, spin-dried, and separated and purified by pre-TLC (DCM:MeOH=10:1) to obtain a white solid 10 (80.3 mg, 47.27 mg) % yield, 96.39% purity).

ESI-MS m/z: 600.35 [M+H] ⁺ .

Example 11: Compound 11 (1-(7-(8-methoxy-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl) )-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one) synthesis

Step 1: Synthesis of Compound 11-1

Under nitrogen protection, Sphos Pd G2 (59.2 mg) was added to compound 10-2 (300 mg), K ₃ PO ₄ (261.5 mg), vinyl boronate (126.53 mg) in dioxane (4 mL), H ₂ O (1 mL) mixed solution was stirred at 100° C. for 4 h. The temperature was lowered to room temperature, diluted with water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, spun dry, separated and purified by pre-TLC (DCM:MeOH=10:1) to obtain a white solid 11- 1 (205 mg, 69.13% yield).

ESI-MS m/z: 722.43 [M+H] ⁺ .

Step 2: Synthesis of Compound 11-2

At room temperature, compound 11-1 (205 mg) was dissolved in a mixed solvent of DCM (6 mL) and TFA (3 mL), moved to 40° C. to react for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the solvent to obtain a brown oil crude product 11-2 (152 mg, crude product).

ESI-MS m/z: 538.32 [M+H] ⁺ .

Step 3: Synthesis of Compound 11

Under nitrogen protection, a solution of acryloyl chloride (25.4 mg) in THF (1 mL) was added to a solution of compound 11-2 (152 mg), THF (4 mL), saturated Na ₂ CO ₃ (1 mL) in an ice-water bath, maintaining the temperature Stir for 5 minutes. It was diluted with water, extracted three times with EA, the organic phases were combined and dried over anhydrous Na ₂ SO ₄ , filtered, spun dry, and separated and purified by pre-TLC (DCM:MeOH=10:1) to obtain 11 as a yellow solid (87.2 mg, 52.1 %Yield).

ESI-MS m/z: 592.43 [M+H] ⁺ .

¹ H NMR (500MHz, Chloroform-d) δ7.90(s, 1H), 7.52(s, 1H), 7.47(d, J=8.5Hz, 1H), 7.36(d, J=8.5Hz, 1H), 6.39(dd,J＝17.0,2.0Hz,1H),6.22(m,2H),5.72(dd,J＝10.2,2.0Hz,1H),5.62(d,J＝17.4Hz,1H),5.03(d , J=11.0Hz, 1H), 4.03(s, 2H), 3.93(s, 2H), 3.75(m, 8H), 3.00(m, 3H), 2.35(s, 3H), 2.14(s, 3H) ,2.11(d,J＝7.1Hz,5H),2.04(m,4H).

Example 12: Compound 12 (1-(7-(2-(1-acetylpiperidin-4-yl)-7-(5-methyl-1H-indazol-4-yl)-8-(2, 2,2-Trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one )Synthesis

Step 1: Synthesis of Compound 12-1

1-Acetylpiperidine-4-carbonyl chloride (567 mg) was added to compound M1 (300 mg) in THF (10 mL) at room temperature, moved to 40°C and stirred for 4 hours. After the reaction was completed, the THF was directly concentrated to obtain the target product 12-1 (460 mg, crude product) as a yellow solid.

ESI-MS m/z: 512.14 [M+H] ⁺ .

Step 2: Synthesis of Compound 12-2

At room temperature, sodium methoxide (224.56 mg) was added to toluene (10 mL) of compound 12-1 (460 mg), and the mixture was moved to 110° C. and stirred under reflux for 8 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to give the target product 12-2 (210 mg, crude) as a white solid.

ESI-MS m/z: 494.06 [M+H] ⁺ .

Step 3: Synthesis of Compound 12-3

Compound 12-2 (170 mg) was added to DMF (5 mL) at room temperature, followed by tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (92 mg), BOP (376 mg) ), DBU (258mg), reacted at 40 degrees for 6h. The reaction was not carried out until 30% of the raw materials remained in the later stage, water was added to the reactant, extracted with ethyl acetate, the organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo, and the concentrate was passed through pre-TLC (developing solvent: DCM/MeOH= 15:1) Purification to yield product 12-3 as a yellow solid (150 mg, 62.2% yield).

ESI-MS m/z: 702.28 [M+H] ⁺ .

Step 4: Synthesis of Compound 12-4

Under nitrogen protection, trifluoroethanol (106 mg) was added to compound 12-3 (150 mg), Cs ₂ CO ₃ (139 mg, ) in dioxane (3 mL), moved to 100° C. to react for 3 h. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, the concentrate was purified by pre-TLC (developing solvent: DCM/MeOH=15:1), The desired target product 12-4 was obtained as a yellow oil (120 mg, 71.9% yield).

ESI-MS m/z: 782.48 [M+H] ⁺ .

Step 5: Synthesis of Compound 12-5

Under nitrogen protection, Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (12 mg) was added to compound 12-4 (115 mg), K ₂ CO ₃ (41 mg, ), 4,4,5,5-tetramethyl A mixed solution of -2-vinyl-1,3,2-dioxaborolane (22.7 mg) in dioxane (2.5 mL) and H ₂ O (0.5 mL) was stirred at 70 °C for 6 h . The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (PE:EA=1: 2) Purification gave the desired target product 12-5 as a yellow solid (65 mg, 65.4% yield).

ESI-MS m/z: 682.58 [M+H] ⁺ .

Step 6: Synthesis of Compound 12-6

Under nitrogen protection, Pd(PPh ₃ ) ₄ (22 mg) was added to compound 12-5 (65 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (65 mg), _K3PO4 (40 mg) in dioxane ( ₂ mL), _H2O (0.5 mL) ) solution, moved to 85°C and reacted for about 4 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo, the concentrate was passed through pre-TLC (PE:EA=2:1 ) to give the desired product 12-6 as a yellow oil (70 mg, 89.7% yield).

ESI-MS m/z: 818.94 [M+H] ⁺ .

Step 7: Synthesis of Compound 12-7

Compound 14-6 (70 mg, 85.7 μmol) was dissolved in a mixed solvent of DCM (1 mL) and TFA (0.5 mL) at room temperature, moved to 40° C. to react for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the solvent to obtain the target product 12-7 (93 mg, crude product) as a yellow solid.

ESI-MS m/z: 634.32 [M+H] ⁺ .

Step 8: Synthesis of Compound 12

Under nitrogen protection, in an ice-water bath, a solution of acryloyl chloride (8.4 mg, 94 μmol) in THF was added to a solution of compound 14-7 (54 mg), THF (2 mL), saturated Na ₂ CO ₃ (0.5 mL), and a solution of The reaction is complete. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM/MeOH=10:1) Purification gave the desired product 12 as a yellow solid (25.7 mg, 43.2% yield).

ESI-MS m/z: 688.49 [M+H] ⁺ .

Example 13: Compound 13 (1-(7-(6-cyclopropyl-8-(2-methoxyethoxy)-7-(5-methyl-1H-indazol-4-yl)- 2-(1-Methylpiperidin-4-yl)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1-one )Synthesis

Step 1: Synthesis of Compound 13-1

Under nitrogen protection, Pd(dppf) _2Cl2DCM (112 _mg ) was added to potassium carbonate (378 mg), compound 1-6 (1 g) and cyclopropylboronic acid (124 mg) in toluene (10 mL) and water (1 mL) The solution was stirred at 100 degrees for 3 hours, TLC and LCMS showed that the reaction was complete, cooled to room temperature, diluted with ethyl acetate, washed twice with saturated brine, dried, filtered, and concentrated to obtain the crude product, which was subjected to column chromatography (DCM/MeOH= 15:1) to obtain the desired product (530 mg, 60% yield). ESI-MS m/z: 668.30[M+H]+

Step 2: Synthesis of Compound 13-2

Under nitrogen protection, Pd2(dba) _{3 (} 9.9 mg) and SPhos (8.8 mg) were added to 13-1 (72 mg),5-methyl-1-(tetrahydro-2H-pyran-2-yl )-4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-1H-indazole (37mg) and potassium phosphate (46mg) in dioxygen Hexacyclic (3mL) and water (0.4mL) solution, heated to 110 degrees and reacted for 2 hours, TLC and LCMS showed that the reaction was complete, cooled to room temperature, concentrated, column chromatography (DCM/MeOH=15:1) to obtain the product (38 mg, 45% yield). ESI-MS m/z:804.50[M+H]+

Step 3: Synthesis of Compound 13-3

At room temperature, 13-2 (200 mg) was dissolved in a mixed solvent of DCM (4 mL) and TFA (2 mL), moved to 40° C. and reacted for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the solvent to obtain the crude target product 13-3 (100 mg) as a yellow solid. ESI-MS m/z: 620.38[M+H]+.

Step 4: Synthesis of Compound 13

Under nitrogen protection, a solution of acryloyl chloride (22 mg) in THF was added to a solution of 13-3 (100 mg) in THF (4 mL) and saturated Na2CO3 (1 mL) in an ice-water bath, and the reaction was complete. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM/MeOH=10:1) Purification gave the desired product 13 as a yellow solid (24.1 mg, 22% yield). ESI-MS m/z: 674.40 [M+H]+.

Example 14: Compound 14 (1-(7-(6-Cyclopropyl-8-(2-methoxyethoxy)-7-(5-methyl-1H-indazol-4-yl)- 2-(1-Methylpiperidin-4-yl)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1-one )Synthesis

Under nitrogen protection, ethylene glycol methyl ether (762 mg) was added to 1-5 (1.35 g), Cs ₂ CO ₃ (1.96 g) in Dioxane (15 mL), and moved to 100° C. to react for 30 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo to give the desired target product 14-1 as a yellow solid (1.3 g , the crude product). ESI-MS m/z: 730.15[M+H]+

Then, 14-2, 14-3 and 14-4 were synthesized according to the synthesis steps of compound 13, and the target product 14 was finally obtained. ESI-MS m/z: 650.4[M+H]+

Example 15: Compound 15 (1-(7-(6-cyclopropyl-8-(2,2-difluoroethoxy)-7-(5-methyl-1H-indazol-4-yl) -2-(1-Methylpiperidin-4-yl)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1- ketone) synthesis

Step 1: Synthesis of Compound 15-1

Under nitrogen protection, difluoroethanol (201 mg) was added to 1-5 (550 mg), Cs ₂ CO ₃ (531.45 mg) in Dioxane (5 mL), moved to 100° C. to react for 30 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo to give the desired target product 15-1 as a yellow solid (550 mg, crude). ESI-MS m/z: 736.30[M+H]+

Then, 15-2, 15-3 and 15-4 were synthesized according to the synthesis steps of compound 13, and the target product 15 was finally obtained. ESI-MS m/z: 656.4[M+H]+

Example 16: Compound 16 (1-(7-(8-ethoxy-6-ethyl-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperin) Synthesis of pyridin-4-yl)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1-one)

Step 1: Synthesis of Compound 16-1

Under nitrogen protection, ethanol (112 mg) was added to 1-5 (550 mg), Cs ₂ CO ₃ (531 mg) in Dioxane (5 mL), moved to 100° C. to react for 30 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo to give the desired target product 16-1 as a yellow solid (590 mg, crude). ESI-MS m/z: 700.3 [M+H]+.

Step 2: Synthesis of Compound 16-2

Under nitrogen protection, Pd(dppf)Cl ₂ .CH ₂ Cl ₂ (67 mg) was added to 16-1 (crude 590 mg), K ₂ CO ₃ (226 mg), 4,4,5,5-tetramethyl- A mixed solution of 2-vinyl-1,3,2-dioxaborolane (126 mg) in dioxane (5.0 mL) and H ₂ O (0.5 mL) was stirred at 70° C. for 1 h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH = 10:1) purification gave the desired product 16-2 as a yellow solid (329 mg, 66% yield). ESI-MS m/z: 600.6 [M+H]+.

Step 3: Synthesis of Compound 16-3

Under nitrogen protection, Pd(PPh ₃ ) ₄ (116.16 mg) was added to 16-3 (329 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (344.04 mg), dioxane ( ₃ mL) in _K3PO4 (213.38 mg), _H2O (0.75 mL) The solution was moved to 85°C and reacted for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH=10:1 ) was purified to give the desired product 16-3 (244 mg, 65% yield) as a yellow solid. ESI-MS m/z: 736.60 [M+H]+.

Step 4: Synthesis of Compound 16-4

At room temperature, 16-3 (100 mg) was dissolved in methanol, 300 mg of palladium on carbon was added, hydrogen was replaced, and the reaction was moved to 40° C. overnight. After the reaction was completed, diatomaceous earth was used for filtration, and the filtrate was concentrated to remove the solvent to obtain the crude target product 16-4 (150 mg, crude) as a yellow solid. ESI-MS m/z: 738.60 [M+H]+.

Step 5: Synthesis of Compound 16-5

At room temperature, 16-4 (150 mg, crude) was dissolved in a mixed solvent of DCM (4 mL) and TFA (2 mL), moved to 40° C. to react for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the solvent to obtain the crude target product 16-5 (100 mg, crude) as a yellow solid. ESI-MS m/z: 554.60 [M+H]+.

Step 6: Synthesis of Compound 16

Under nitrogen protection, the THF solution of acryloyl chloride (17.03 mg) was added to 16-5 (100 mg), THF (4 mL), saturated Na2CO3 (1 mL) solution in an ice-water bath, and the reaction was completed after the addition. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM/MeOH=10:1) Purification gave the desired product 16 as a yellow solid (21 mg, 20% yield). ESI-MS m/z: 608.40 [M+H]+.

Embodiments 17-22 respectively adopt cyclobutanol, isopropanol, difluoroethanol, 2-fluoroethanol, ethylene glycol methyl ether and cyclopropyl methanol to react with compounds 1-5 to obtain corresponding products, and then follow the synthetic steps of 16 The corresponding final product compounds 17-22 were synthesized.

Example 23: Compound 23 (1-(7-(6-Chloro-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl)- 8-(2,2,2-Trifluoroethoxy)quinazoline-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-ene-1- ketone) synthesis

Step 1: Synthesis of Compound 23-1

To a solution of 2-amino-4-bromo-3-fluorobenzoic acid (20 g) in DMF (200 mL) was added NCS (13.8 g) at room temperature and the resulting mixture was stirred at 70 °C overnight. The reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo to give the desired crude product as a brown solid (22 g, 98% yield). ESI-MS m/z: 267.10 [M+H]+.

Step 2: Synthesis of Compound 23-2

1-1 (2.42 g) was added to M1-5 (2.33 g) in THF (20 mL) at room temperature, moved to 40°C and stirred overnight. After the reaction is complete, add 30 mL of ammonia water, adjust the pH of the solution to 11, stir at 35 degrees, detect until the reaction is complete, a large amount of insoluble matter is precipitated, and the insoluble matter is filtered to obtain the crude product 23-2 (3.5g, crude). ESI-MS m/z: 374.2 [M+H]+.

Step 3: Synthesis of Compound 23-3

Under nitrogen protection at room temperature, compound 23-2 (1 g) and tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (725 mg) were added to the reaction flask, DMF (10 mL) and DBU were added (2.44g), cooled in an ice-water bath, added BOP (2.95g), warmed up to 45 degrees and reacted for 2 hours, detected the completion of the reaction, cooled to room temperature, diluted with ethyl acetate, diluted with water, extracted once with ethyl acetate in the aqueous phase, and the organic phase Combined, washed 3 times with 0.5N hydrochloric acid and Brine (1:1), once with aq. NaHCO ₃ , once with brine, dried over sodium sulfate, and concentrated to obtain the target product 23-3 (1.2 g, crude product).

Referring to the synthesis steps of compound 13, 23-4, 23-4 were synthesized by substituting the fluorine atom with trifluoroethanol under basic conditions. 23-5 and 23-5 were synthesized by the method of Suzuki coupling. To obtain compound 23-6, 23-6 was reacted with acryloyl chloride under alkaline conditions to synthesize the target product 23.

The synthesis of compounds 24 and 25 was completed with reference to the synthesis method of compound 23, 24 and 25 were respectively replaced by cyclobutanol and difluoroethanol instead of trifluoroethanol to replace the fluorine atom to obtain the corresponding products, and then the target products 24 and 25 were synthesized according to the steps of 23.

Example 26: Compound 26 (1-(7-(6-ethyl-2-(1-(2-methoxyethyl)piperidin-4-yl)-7-(5-methyl-1H- Indazol-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl) Synthesis of prop-2-en-1-one)

Step 1: Synthesis of Compound 26-1

At room temperature, piperidine-4-carboxylic acid (7.000 g) was added to THF (200 mL) and water (40 mL), triethylamine (16.45 g) was added, and (2,5-dioxapyrrolidine-1- base) 2-trimethylsilyl ethyl carbonate (15.46g), stirred at room temperature until the reaction was completed, concentrated THF, cooled, acidified to pH2-3 with 1N HCl, extracted three times with DCM, washed twice with organic phase brine , dried and concentrated to obtain the target product 26-1 (16 g, crude product).

Step 2: Synthesis of Compound 26-2

At room temperature, compound 26-1 (2.73 g) was added to DCM (30 mL), 2 drops of DMF were added, oxalyl chloride (3.81 g) was added dropwise, bubbles appeared, the reaction was completed, and concentrated to obtain 26-2 (3 g, crude product), used directly in the next step.

Step 3: Synthesis of Compounds 26-3 and 26-4

At room temperature, 26-2 (3 g) was added to THF (40 mL) of M1 (1.8 g), moved to 40 °C and stirred overnight, and detected until the reaction was complete to obtain 26-3, added 30 mL of ammonia water, and adjusted the pH of the solution to Stir at 11, 40 degrees, detect until the reaction is complete, a large amount of insoluble matter is precipitated, and the insoluble matter is filtered to obtain 26-4 (1.9 g, 63% yield). ESI-MS m/z: 374.2 [M+H]+.

Step 4: Synthesis of Compound 26-5

Under nitrogen protection at room temperature, compound 26-4 (1.22 g) and tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (695 mg) were added to the reaction flask, DMF (12 mL) and DBU (1.87g) was cooled in an ice-water bath, BOP (2.2g) was added, the temperature was raised to 45°C for 2 hours, the reaction was detected, cooled to room temperature, diluted with ethyl acetate, diluted with water, extracted once with ethyl acetate in the aqueous phase, and the organic The phases were combined, washed 3 times with 0.5N hydrochloric acid and Brine (1:1), once with aq. NaHCO ₃ , once with Brine, dried over sodium sulfate, and concentrated to obtain the target product 26-5 (1 g, crude product).

Step 5: Synthesis of Compound 26-6

Under nitrogen protection, trifluoroethanol (2.77 g) was added to 26-5 (2.23 g), Cs ₂ CO ₃ (2.7 g) in Dioxane (25 mL), moved to 100° C. to react for 30 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo to give the desired desired product 26-6 as a solid (4.6 g, crude). ESI-MS m/z: 784.30[M+H]+

Step 6: Synthesis of Compound 26-7

Under nitrogen protection, Pd(dppf) _2Cl2DCM ( _1.82g ) was added to the dioxane of potassium carbonate (6.16g), compound 26-6 (19.7g) and vinylborate (3.43g) (200 mL) and water (20 mL), stirred at 100 degrees for 3 hours, TLC and LCMS showed that the reaction was complete, cooled to room temperature, diluted with ethyl acetate, washed twice with saturated brine, dried, filtered, and concentrated to obtain the crude product, Column chromatography gave (DCM/EA=20:1) the desired product (9.6 g, 55% yield). ESI-MS m/z: 895.33[M+H]+

Step 7: Synthesis of Compound 26-8

Under nitrogen protection, Pd(PPh ₃ ) ₄ (2.94 g) was added to D1-7 (10 g), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5, A solution of 5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (7.85 g) and potassium phosphate (5.41 g) in dioxane (100 mL) and water (20 mL) , heated to 85 degrees and reacted for 4 hours, TLC and LCMS showed that the reaction was completed, cooled to room temperature, diluted with ethyl acetate, washed with water, washed with saturated brine, dried and concentrated to obtain the crude product, and column chromatography gave a yellow foamy solid (10 g, 85 % yield). ESI-MS m/z: 920.40[M+H]+

Step 8: Synthesis of Compound 26-9

At room temperature, 26-8 (1 g) was dissolved in methanol, palladium carbon (1 g) was added, hydrogen was replaced, and the reaction was moved to 40° C. overnight. After the reaction was completed, diatomaceous earth was used for filtration, and the filtrate was concentrated to remove the solvent to obtain the crude target product 26-9 (1 g, crude) as a yellow solid. ESI-MS m/z: 922.50 [M+H]+.

Step 9: Synthesis of Compounds 26-10

At room temperature, compound 26-9 (1.6 g) was dissolved in THF (10 mL), TBAF (4 mL, 1M in THF) was added, stirred at 35 degrees for 4 hours, concentrated, diluted with ethyl acetate, washed twice with water, and washed with saturated brine twice, dried over sodium sulfate, and concentrated to give the product (912 mg, 67% yield).

Step 10: Synthesis of Compounds 26-11

Compound 26-10 (100 mg) was dissolved in DMF (5 mL) at room temperature, cesium carbonate (126 mg) was added, 2-bromoethyl methyl ether (54 mg) was added, and the mixture was stirred overnight at room temperature. The reaction was monitored by LCMS and diluted with ethyl acetate. Washed once with water, washed twice with saturated brine, dried, concentrated to obtain crude product, prepared by plate (DCM/MeOH=10:1) to prepare 26-11 (37 mg, 34% yield) ESI-MS m/z: 836.46 [M +H]+

Step 11: Synthesis of Compounds 26-12

At room temperature, 26-11 (37 mg) was dissolved in a mixed solvent of DCM (2 mL) and TFA (1 mL), moved to 40° C. to react for about 1 h. After the reaction is complete, the solvent is directly concentrated to remove the solvent to obtain the crude target product 26-12. ESI-MS m/z: 652.40 [M+H]+.

Step 12: Synthesis of Compound 26

Under nitrogen protection, in an ice-water bath, a solution of acryloyl chloride (4 mg) in THF was added to a solution of 26-12 (29 mg, crude product) in THF (2 mL) and saturated Na ₂ CO ₃ (0.5 mL), and the reaction was complete after the addition. . The reaction mixture was poured into water, extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was purified by pre-TLC (DCM/MeOH=10:1) to give the desired product 26 (3.1 mg, 6% yield). ESI-MS m/z: 706.45 [M+H]+.

Example 27: Compound 27 (1-(7-(6-ethyl-2-(1-(2-hydroxy-2-methylpropyl)piperidin-4-yl)-7-(5-methyl) -1H-Indazol-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2 Synthesis of -yl)prop-2-en-1-one)

Step 1: Synthesis of Compounds 27-11

Compound 27-10 (100 mg, 0.13 mmol) was dissolved in ethanol (2 mL), methyl propylene oxide (93 mg, ) and triethylamine (40 mg) were added, and the reaction was carried out by microwave at 100 degrees for 0.5 hours. After the reaction was detected by LCMS, it was directly concentrated. Obtained crude 27-11.

Referring to the synthetic method of Example 26, the protecting group was removed under the action of trifluoroacetic acid, and then reacted with acryloyl chloride under basic conditions to obtain the target product 27 (8.8 mg). ESI-MS m/z: 720.35[M+H]+

Example 28: Compound 28 (1-(7-(6-ethyl-7-(5-methyl-1H-indazol-4-yl)-2-(1-(oxetan-3-yl) )piperidin-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl ) synthesis of prop-2-en-1-one)

Step 1: Synthesis of Compounds 28-11

Compound 26-10 (165 mg) was dissolved in 1,2-dichloroethane (10 mL), 3-oxetanone (76.4 mg,) and sodium borohydride acetate (225 mg) were added, and the mixture was stirred at room temperature at 35 degrees overnight. The reaction was completed, diluted with ethyl acetate, washed with water, washed with saturated brine, dried and concentrated to obtain the crude product.

Referring to the synthesis steps of Example 26, the protective group was removed under the action of trifluoroacetic acid, and then reacted with acryloyl chloride under basic conditions to obtain the target product 28 (13.5 mg). ESI-MS m/z:704.50[M+H]+

Example 29: Compound 29 (1-(7-(6-ethyl-2-(1-(2-hydroxy-2-methylpropyl)azetidin-3-yl)-7-(5 -Methyl-1H-indazol-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nona Synthesis of yl-2-yl)prop-2-en-1-one).

See Examples 26 and 27 for synthetic procedures.

Operation of compound 29-1 Referring to the synthesis operation of compound 26-1, the nitrogen atom of 3-azetidinecarboxylic acid was protected. Operation of compound 29-2 Referring to the synthesis operation of compound 26-2, the carboxylic acid of compound 26-2 was converted into an acid chloride, and then reacted with compound M1 to obtain compound 29-3. Compound 29-3 was directly added with ammonia water without treatment to adjust the pH to 11 ring closure to synthesize compound 29-4. Compound 29-4 was introduced into compound 29-5 by spiro ring under the action of DBU and BOP, and then its fluorine atom was replaced with trifluoroethanol to obtain compound 29-6. Compound 29-6 was subjected to two Suzuki coupling reactions to synthesize compound 29-8 , and then the double bond is reduced, and the protecting group of the nitrogen atom is removed to obtain the shared intermediate 29-10. Compound 29-10 can be obtained by microwave reaction with methyl propylene oxide in triethylamine and ethanol system with reference to the synthesis steps of 27-11, followed by deprotection of trifluoroacetic acid to obtain compound 29-12. Reaction with acryloyl chloride under neutral conditions affords compound 29.

Example 30: Compound 30 (3-(3-(4-(2-Acryloyl-2,7-diazaspiro[3.5]nonan-7-yl)-6-ethyl-7-(5 -Methyl-1H-indazol-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-2-yl)azetidin-1-yl)propionitrile) Synthesis

Step 1; Synthesis of compounds 30-11

Compound 29-10 (50 mg) was dissolved in ethanol (3 mL), acrylonitrile (34 mg) and triethylamine (7 mg) were added, the reaction was carried out at room temperature 35 degrees for 4 hours, concentrated, diluted with ethyl acetate, washed with water, and washed with saturated brine , dried and concentrated, and purified by preparative plate (DCM/MeOH=20:1) to give the product (26 mg). ESI-MS m/z:830.20[M+H]+

Then, referring to the synthetic method of Example 26, the protecting group was removed under the condition of trifluoroacetic acid, and the acryloyl group was introduced under the basic condition to obtain compound 30.

Examples 31, 33, 34, 35 and 37 were synthesized with reference to the method of Example 28, sharing intermediates 29-10 with oxetanone, tetrahydropyranone, cyclobutanone, cyclopentanone and acetaldehyde, respectively. The corresponding target compounds were prepared under the conditions of reductive amination, and then deprotected and acryloyl groups were introduced to obtain target compounds 31, 33, 34, 35 and 37 respectively.

The synthetic operation of embodiment 32 refers to the synthetic operation of embodiment 30, adopts the method of 1,4-addition, methyl vinyl sulfone reacts with common intermediate 29-10 to obtain the corresponding product, and then deprotection and introduction of acryloyl group finally obtain the target Compound Example 32.

The synthetic procedure of embodiment 36 is: use common intermediate 29-10 and cyclopropylcarbonyl chloride to react in dichloromethane and sodium bicarbonate aqueous solution system to obtain corresponding compound, then deprotect and introduce acryloyl group to finally obtain target compound embodiment 36.

Example 38: Compound 38 (1-(7-(8-(2-hydroxy-2-methylpropoxy)-7-(5-methyl-1H-indazol-4-yl)-2-( 1-Methylpiperidin-4-yl)-6-vinylquinazoline-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-ene-1 -ketone) synthesis

Synthetic operation of Example 38 Referring to the operation of Example 16, 2-methyl-1,2-propanediol was used to react with the common intermediate 1-5 to obtain compound 38-1, and then two coupling reactions were carried out according to Example 16, Deprotection and acryloylation steps complete the synthesis of 38.

Examples 39 and 40 were respectively reacted with difluoroethanol and 1-(hydroxymethyl)cyclopropanecarbonitrile with the common intermediates 1-5 to obtain the corresponding target compounds, and then the final products 39 and 40 were prepared according to the steps of Example 16. synthesis

Example 41: Compound 41 (1-(6-(7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl)-8-(2 ,2,2-Trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,6-diazaspiro[3.3]hept-2-yl)prop-2-ene-1- ketone) synthesis

Step 1: Synthesis of Compound 41-1

DIEA (480 mg) was added to a solution of 1-4 (600 mg), 2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (245 mg) in dioxane (10 mL) at room temperature , stirring at room temperature 40 degrees. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the crude product was purified by flash silica gel column chromatography (DCM:MeOH=10:1) to give a yellow color The desired target product 41-1 as a solid (680 mg, 85% yield).

After that, the synthesis of the target product 41 was completed with reference to the synthesis steps of Example 16.

For the synthesis of Examples 42, 43 and 44, refer to the synthesis of 41-1 using 2,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester, (S)-4-N-tert-butoxy Carbonyl-2-methylpiperazine and tert-butyl(S)-2-(cyanomethyl)piperazine-1-carboxylate react with common intermediates 1-4 to obtain the corresponding compounds, followed by the synthesis of reference 25 The steps complete the synthesis of target products 42, 43 and 44.

Example 45: Compound 45 (1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(1-(2-(methylsulfonyl)ethyl)piperidine -4-yl)-8-(2,2,2-trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2 Synthesis of -yl)prop-2-en-1-one)

Step 1: Synthesis of Compound 45-1

At room temperature, compound 26-8 (340 mg) was dissolved in THF (5 mL), TBAF (2 mL, 1 M in THF) was added, stirred at 35 degrees overnight, concentrated, diluted with ethyl acetate, washed twice with water and twice with saturated brine , dried over sodium sulfate, and concentrated to give the product (311 mg, crude product).

Step 2: Synthesis of Compound 45-2

Compound 45-1 (50 mg) was dissolved in ethanol (3 mL) at room temperature, methyl vinyl sulfone (68 mg) was added, stirred overnight at room temperature, monitored by LCMS, the reaction was completed, concentrated, diluted with ethyl acetate, washed once with water, and washed with saturated brine for two times. times, dried and concentrated to obtain crude product, which was prepared by plate preparation (DCM/MeOH=20:1) to obtain 45-2 (32 mg, 36% yield). ESI-MS m/z: 882.46[M+H]+

Step 3: Synthesis of Compound 45-3

At room temperature, 45-2 (32 mg) was dissolved in a mixed solvent of DCM (2 mL) and TFA (1 mL), moved to 40° C. and reacted for about 1 h. After the reaction was completed, the solvent was directly concentrated to remove the solvent to obtain the crude target product 45-3. ESI-MS m/z: 698.40 [M+H]+.

Step 4: Synthesis of Compound 45

Under nitrogen protection, a solution of acryloyl chloride (4 mg) in THF was added to a solution of 45-3 (25 mg crude product) in THF (2 mL) and saturated Na ₂ CO ₃ (0.5 mL) in an ice-water bath, and the reaction was complete. The reaction mixture was poured into water, extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo, the concentrate was purified by pre-TLC (DCM/MeOH=10:1) to give the desired product 45 (6.6 mg, 24% yield). ESI-MS m/z: 752.45 [M+H]+.

Example 51: Compound 51 (1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(piperidin-4-yl)-8-(2,2,2 -Trifluoroethoxy)-6-vinylquinazoline-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1-one) of synthesis

Step 1: Synthesis of Compound 51-1

Compound 26-8 (100 mg) was dissolved in dichloromethane (2 mL), hydrochloric acid dioxane solution (0.5 mL, 4N in dioxane) was added, and the reaction solution was concentrated to obtain a crude product (50 mg), which was directly used in the next step.

Step 2: Synthesis of Compound 51-2

Under nitrogen protection, a solution of acryloyl chloride (6 mg) in THF was added to a solution of 51-1 (50 mg, crude) in THF (2 mL) and saturated Na ₂ CO ₃ (1 mL) in an ice-water bath, and the reaction was complete. The reaction mixture was poured into water, extracted with ethyl acetate, the organic layer was washed with brine, dried _{over Na2SO4} and concentrated in vacuo to give crude product (30 mg).

Step 3: Synthesis of Compound 51

Compound 51-2 (30 mg) was dissolved in dichloromethane (1 mL), TFA (1 mL) was added, stirred at room temperature for 2 hours, the reaction was completed, concentrated, and the target product (2.6 mg, 11% yield).

Example 47 is synthesized with reference to the steps of Example 1, and the corresponding target product can be obtained by reacting acrylonitrile with the common intermediate 45-1, followed by deprotection and introduction of an acryloyl group to finally obtain the target compound 47.

Examples 46, 53, 54, 55, 56, 57, 59, 60, 61, 62, 65, 66, 67, 68, 70 and 71 refer to Example 28 by reductive amination using different ketones or aldehydes Reaction with shared intermediate 45-1 to obtain the corresponding product, then deprotection and introduction of acetyl group to synthesize the final target product; Embodiment 48 refers to Example 27, and uses shared intermediate 45-1 to react with methyl propylene oxide to generate corresponding The product, then deprotection and introduction of an acryloyl group to finally obtain the target compound 48; Examples 49, 58, 63, 64, 69 adopt an alkylation strategy to react with the common intermediate 45-1 to generate the corresponding product, and then deprotection and The introduction of the acryloyl group finally yields the final target product. The corresponding synthetic intermediates are as follows:

Example 72: Compound 72 (2-fluoro-1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl)- 8-(2,2,2-Trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)propan-2 -en-1-one) synthesis

Compound 2-8 (120 mg) was dissolved in DMF (5 mL), 2-fluoroacrylic acid (54 mg), HATU (226 mmol), DIEA (77 mg) were added, stirred for 2 hours, the reaction was completed, diluted with ethyl acetate, washed with saturated brine Three times, dried and concentrated to obtain the crude product, and the target product (3 mg) was obtained by preparative plate preparation (DCM/MeOH=10/1).

Example 75: Compound 75 ((E)-1-(7-(6-Chloro-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidine-4 -yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)-4-( Synthesis of dimethylamino)but-2-en-1-one)

A freshly prepared solution of trans-4-dimethylaminocrotonyl chloride hydrochloride (12 mg) in THF was added to 23-6 (100 mg, crude) in THF (2 mL) and Saturated Na ₂ CO ₃ (1 mL) solution was added to complete the reaction. The reaction mixture was poured into water, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo to give crude product, which was prepared as a plate (DCM/MeOH=10:1) (6 mg, 5 % yield).

The synthesis of Example 73 refers to the synthesis procedure of 50, except that difluoroethanol is used instead of trifluoroethanol.

The synthesis of Example 74 refers to the synthetic procedure of 72.

Referring to Example 1A, the compounds of Examples 2 to 75 were resolved by substantially the same method, and the corresponding chiral isomers 2a to 75a were obtained respectively. For example, the structure of compound 9a is

The structural formula of compound 47a is

Synthesis of Comparative Compound 1 (D1)

For the synthesis of AMG510(D1), refer to WO2018217651A1.

Synthesis of Comparative Compound 2 (D2)

D2 synthesis reference WO2018143315A1.

Pharmacological experiments

Example A: Cell Proliferation Inhibition Experiment

The CALU-1 cells were plated in a 96-well ultra-low adsorption plate at 1000 cells and 190 μL/well. After overnight incubation, compound solutions with gradient concentrations were prepared, and 10 μL of the DMSO solution of each concentration of the test compound was added to the cells in each well. 0 nM (all final concentrations of DMSO were 0.25%). Incubate for 120 hours at 37°C, 5% _CO2 . Add 60 μL of Cell-titer Glo working solution to each well, shake and mix well, incubate at room temperature for 10 minutes, read the Luminescence luminescence value with a multi-function microplate reader, and convert the luminescence value reading into inhibition percentage:

Percent Inhibition=(Max-Reading)/(Max-Min)*100.

"Max" is the DMSO control; "Min" is the cell-free control.

Curve fitting was performed with Graphpad Prism software and _IC50 values were obtained.

Example compounds have good inhibitory activity on CALU-1 cells, especially the Y substituent is selected from

Compared with other ring structure molecules directly connected to the quinazoline ring through the C atom on the ring, it has better activity. See Table 2 for _IC50 data of the compounds of the examples for inhibition of CALU-1 cells.

Table 2

Compound name	IC50 (nM)
Compound 1	2.7
Compound 2	776.1
Compound 3	277.6
Compound 4	2.9
Compound 5	45.7
Compound 6	4.6

Example B: Cell Proliferation Inhibition Experiment

MIA-PACA-2 cells were plated in a 96-well ultra-low adsorption plate at 600 cells and 160 μL/well. After overnight incubation, compound solutions with gradient concentrations were prepared, and 40 μL of DMSO solution of each concentration of the test compound was added to each well of cells respectively. 0 nM (all final concentrations of DMSO were 0.25%). Incubate for 96h at 37°C, 5% _CO2 . Add 50 μL of Cell-titer Glo working solution to each well, incubate at room temperature for 10 min after shaking and mixing, read the Luminescence luminescence value with a multi-function microplate reader, and convert the luminescence value reading into the inhibition percentage:

Percent Inhibition=(Max-Reading)/(Max-Min)*100.

"Max" is the DMSO control; "Min" is the cell-free control.

Curve fitting was performed with Graphpad Prism software and _IC50 values were obtained.

Table 3 shows the _IC50 data of some compounds in the examples for inhibition of MIA-PACA-2 cells.

Example C: Cell Proliferation Inhibition Experiment

H358 cells were plated in 96-well ultra-low adsorption plates at 2000 cells and 190 μL/well. After overnight incubation, compound solutions with gradient concentrations were prepared, and 10 μL of DMSO solution of each concentration of the test compound was added to each well of cells respectively. 0 nM (all final concentrations of DMSO were 0.25%). Incubate for 96h at 37°C, 5% _CO2 . Add 50 μL of Cell-titer Glo working solution to each well, incubate at room temperature for 10 min after shaking and mixing, read the Luminescence luminescence value with a multi-function microplate reader, and convert the luminescence value reading into the inhibition percentage:

Percent Inhibition=(Max-Reading)/(Max-Min)*100.

"Max" is the DMSO control; "Min" is the cell-free control. Curve fitting was performed with Graphpad Prism software and _IC50 values were obtained.

Table 3 shows the _IC50 data of some compounds in the examples for inhibition of H358 cells.

table 3

Example D: Isomer Activity

Use chromatographic conditions: CHIRALCEL OD (2.5cm I.D.×25cm L, 10μm), UV 254nm, mobile phase Hexane/IPA/DEA=70/30/0.1 (V/V/V), flow rate 20ml/min, the compound was separated. Then, the isomer activity was determined using the methods of Example B and Example C.

The results show that the activity of the representative isomer of the chiral compound represented by the general formula (I) of the present invention is better than that of the corresponding axial chiral isomer, and the activity difference is more than 100 times.

Table 4

table 5

Example E: In vivo pharmacodynamic studies on human pancreatic cancer MIA PaCa-2 CDX tumor model

The classical mouse tumor model test was used to observe the tumor inhibition rate TGI (%) of the target compound after oral administration to evaluate its anti-tumor activity.

Experimental methods: Human pancreatic cancer MIA PaCa-2 cells were cultured in monolayer in vitro, cultured in DMEM/F12 medium with 10% fetal bovine serum, 1% double antibody, and cultured in a 37°C 5% CO ₂ incubator. Routine treatment passaging was performed twice a week. When the cell saturation is 80%-90% and the number reaches the requirement, the cells are collected, counted, and seeded. 0.2 mL (5×10 ⁶ cells) of MIA PaCa-2 cells (plus Matrigel, 1:1 by volume) were subcutaneously inoculated into the right back of each mouse, and the tumors were grouped into groups when the average tumor volume reached about 149 mm ³ . medicine.

Experimental results: Both the target compound 1a and AMG510 can effectively reduce tumor TGI, which is significantly different from the solvent control group (Figure 4). The target compound 1a treated mice with better body weight changes and showed better safety potential (Figure 5).

Example F: In vivo pharmacodynamic study of human non-small cell lung cancer PDX tumor model

The classical mouse tumor model test was used to observe the tumor inhibition rate TGI (%) of the target compound after oral administration to evaluate its anti-tumor activity.

Experimental methods: The establishment of human lung cancer LU-01-0030PDX model was originally derived from surgically resected clinical samples of G12C mutation-positive patients, which were defined as P0 generation after implantation in nude mice. The implantation of tumor tissue from the P0 generation into the next generation is called the P1 generation. And so on for continuous implantation in nude mice. The FP3 tumor was recovered from the P2 generation. The next generation from the FP3 generation is designated as FP4, and so on. The tumor tissue of the FP5 generation was subcutaneously inoculated into the right upper limb of each mouse, and the group administration started when the average tumor volume reached about 141 mm3.

Experimental results: On the LU-01-0030PDX model of human non-small cell lung cancer, a weak tumor inhibitory effect was observed with 10mg/kg AMG 510, and the TGI was 3.3%; the target compound 1a was administered with 3mg/kg, 10mg/kg or 30mg/kg , TGI% were 32%, 28% and 52%, the effect was better than AMG 510 (Figure 6). Compound 1a showed good resistance.

Example G: hERG test

1 Steady Transduced Cell Preparation

The cell line was derived from HEK293 cells overexpressing hERG potassium ion channel, and the cells were cultured in a 37°C, 5% CO2 incubator. When the cell density reaches 80% of the dish, pre-wash with phosphate buffered saline (PBS), then digest the cells with trypsin/EDTA for 2-3 minutes, add cell culture medium to stop the digestion, pipette gently and Transfer to a centrifuge tube, centrifuge at 1000 rpm for 3 minutes, pour off the supernatant, add the cell culture medium, mix the cells by gently pipetting, and then transfer to a petri dish for subculture, or drop the cells on a circular The cells were cultured in cell culture medium on glass slides until the cells adhered for the experiments.

Cell culture medium composition: DMEM, 15% fetal bovine serum and 1% 100x penicillin-streptomycin.

2 solution preparation

The composition of intracellular fluid and extracellular fluid is shown in Table 4.

Table 6

3 Electrophysiological Manual Patch Clamp System Experimental Protocol

The stably transfected cells were dropped on a circular glass slide and placed in a petri dish, the cell density was less than 50%, and cultured overnight. The experimental cells were transferred to a cell bath embedded in an inverted microscope platform and perfused with extracellular fluid at a rate of 2.7 ml/min. After 5 minutes of stabilization, the experiment can be started. Membrane currents were recorded using a HEKA EPC-10 patch clamp amplifier and PATCHMASTER acquisition system (HEKA Instruments Inc., D-67466 Lambrecht, Pfalz, Germany). All experiments were performed at room temperature (22-24°C). Electrodes (BF150-110-10) were straightened using a P-97 microelectrode puller (Sutter Instrument Company, One Digital Drive, Novato, CA 94949) in the experiments. The inner diameter of the electrode is 1-1.5mm, and the water entry resistance after filling with the inner liquid is 2-4MΩ.

The electrophysiological stimulation scheme of hERG potassium channel is to first clamp the membrane voltage at -80mV, give the cell a continuous 2s, +20mV voltage stimulation, activate the hERG potassium channel, and then repolarize to -50mV for 5s to generate an outward tail current, The stimulation frequency was every 15s. The current value is the peak value of the tail current.

The channel currents were recorded in the whole-cell recording mode. First perfuse extracellular fluid (about 2 ml per minute) and record continuously, and wait for the current to stabilize (the current decay (Run-Down) is less than 5% within 5 minutes), at which time the peak tail current is the control current value. Then, the extracellular fluid containing the drug to be tested was perfused and recorded continuously until the inhibitory effect of the drug on the hERG current reached a steady state. At this time, the peak value of the tail current was the current value after the drug was added. The standard of steady state is judged by whether the last three consecutive current recording lines overlap. After reaching a stable state, if the hERG current is restored or close to the size before adding the drug after perfusion and flushing with extracellular fluid, you can continue to test other concentrations or drugs by perfusion. 30 μM Quinidine (quinidine) was used in the experiments as a positive control to ensure the normal response of the cells used.

4 Compound Preparation

Dilute the mother liquid with DMSO to the following concentrations (secondary mother liquid), respectively: 3.3, 1.1, 0.37, 0.12 mM, take 30 μl of each of the mother liquid and the secondary mother liquid and dilute into 10 ml of extracellular fluid for electrophysiological detection;

The final concentrations of the compounds were 30, 10, 3.3, 1.1, 0.37 μM;

The final concentration of DMSO was 3:1000.

5 Data Analysis

Experimental data were collected using PATCHMASTER V2X60 (HEKA Instruments Inc., D-67466 Lambrecht, Pfalz, Germany), and analyzed and statistically analyzed using Origin 8.5 (OriginLab Corporation, Northampton, MA) software and Microsoft Excel.

By measuring the maximum current value of the control group and the drug-treated group, calculating the ratio of the maximum current value of the treatment group to the maximum current value of the control group, and evaluating the effect of the test compound on the hERG potassium channel at the test concentration (Mean±SE) .

6Quality Control

This study was performed in accordance with the standard operating procedures of Keruis laboratory (approximately GLP specifications), and all experimental methods were referenced to the literature published in peer-reviewed journals, and were performed in accordance with the standard experimental operating procedures of Keruis Biological.

The test data in the report needs to meet the following criteria:

Electrophysiological recording parameters

Sealing resistance>500MΩ

Direct connection resistance (Rs)<10MΩ

Initial tail current amplitude>300pA

Current rundown (spontaneous decrease) <2%/min

Leakage current <200pA or 10% of peak hERG current (within 90% of recording time)

7 Experimental results:

Table 7

compound	hERG IC 50 (uM)
1a	>30
D2	8.12

8 Experimental conclusions:

Inhibition of cardiac hERG potassium channel by drugs is the main reason for drug-induced QT prolongation syndrome. Compound 1a has almost no blocking effect on hERG potassium channel in the tested concentration range (IC ₅₀ >30μM); Compound D2 has moderate blocking effect on hERG potassium channel in the tested concentration range (3μM<predicted IC ₅₀ <10μM) , the compound has a certain degree of cardiac risk due to its effect on hERG.

Example H: In vitro CYP enzyme inhibition assay

The human liver microsome incubation system was used to reflect the activities of various enzymes by the amount of metabolites generated.

Experimental method: Weigh an appropriate amount of the analyte standard and dilute it to 2 mM with DMSO to obtain a working solution; the probe substrate is prepared with pure acetonitrile, and the positive control inhibitor is prepared with DMSO. The concentrations are shown in Table 8. Take 1 μL of the test compound/positive control inhibitor, 1 μL of the probe substrate, 1 μL of human liver microsomes (20 mg/mL) and 177 μL of PBS buffer to prepare a reaction mixture (the final concentration of the test compound/positive control inhibitor in the incubation reaction is 10 μM ). The above mixture was pre-incubated at 37°C for 5 minutes. 20 μL of NADPH solution (10 mM, prepared in PBS) was added to the reaction mixture, and the CYP2C8 and CYP2C9 enzymes were incubated at 37° C. for 10 minutes, and the CYP2C19 enzymes were incubated for 30 minutes. All incubation samples were double-sampled. The reaction was stopped by adding 200 μL ice-cold acetonitrile containing internal standard. Centrifuge at 3200 rpm for 15 minutes. The supernatant was transferred to LC-MS/MS analysis. The activity of various enzymes is reflected by the production of metabolites. The single-point method is used to calculate, and the formula is as follows:

Inhibiton%=100%*(1-Test compound Aear ratio/Control Aear ratio), the experimental results are shown in Table 9.

Table 8

metabolic enzymes	Probe Substrate/Concentration	Positive Control/Concentration
CYP2C8	Paclitaxel/2mM	Quercertin/2mM
CYP2C9	Diclofenac/1mM	Sulfaphenazole/2mM
CYP2C19	S-Mephenytoin/10mM	(+)-N-3-Benzylnirvanol/2mM

Table 9%Inhibition on major CYP isoforms@10μM

Experimental results: the preferred compounds of the present invention have weak inhibitory effects on CYP2C8, CYP2C9 and CYP2C19 enzymes, so the possibility of drug-metabolic interactions is low in clinical administration.

Although the present invention has been fully described in terms of its embodiments, it is worth noting that various changes and modifications will be apparent to those skilled in the art. Such changes and modifications are intended to be included within the scope of the appended claims of the present invention.

Claims (15)

1. A kind of chiral compound, deuterated substance or pharmaceutically acceptable salt shown in general formula (I),

   

   wherein X is selected from C _3-14 cycloalkyl, 3-14 membered heterocyclyl, 3-14 membered heterocyclylamino; X is optionally further substituted with 1-4 substituents, wherein each substituent is independently Selected from C _1-6 alkyl, amino, C _1-6 aminoalkyl, carbamoyl, C _1-6 carbamoyl alkyl, carboxyl, C _1-6 carboxyalkyl, cyano, C _1-6 cyanoalkyl, halogen, C _1-6 haloalkyl, hydroxyl, C _1-6 hydroxyalkyl;

   R ₁ is acryloyl, substituted acryloyl or

   

   R ₂ is selected from H, C _2-6 alkenyl, C _1-6 alkoxy, C _1-6 haloalkoxy, C _1-6 alkyl, C _1-6 alkane substituted by 4-10-membered heterocyclyl base, C _1-6 alkylsulfonyl, C _1-6 alkylsulfinyl, C _1-6 alkylthio, C _2-6 alkynyl, C _1-6 alkylamino, amino, C _1-6 Aminoalkyl, carbamoyl, C _1-6 carbamoylalkyl, C _1-6 carboxyalkyl, cyano, C _1-6 cyanoalkyl, halogen, C _1-6 haloalkyl, substituted or unsubstituted Substituted aryl, substituted or unsubstituted 5-10 membered heteroaryl, substituted or unsubstituted C _3-7 cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclyl, hydroxy or oxo;

   R ₃ is selected from H, halogen, C _1-6 alkyl, substituted C _1-6 alkyl, C _2-6 alkenyl, substituted C _2-6 alkenyl, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl;

   R ₄ or R ₅ is independently selected from H, halogen, C _1-6 alkyl, substituted C _1-6 alkyl, C _3-6 cycloalkyl, substituted C _3-6 cycloalkyl;

   R ₆ is selected from hydroxyl, oxo, C _1-6 alkoxy, C _3-10 cycloalkyloxy, cyano-substituted cyclopropyl C _1-6 alkyleneoxy, the C _{1- 6} alkoxy is optionally further substituted by one or more substituents selected from halogen, hydroxy, C _1-6 alkoxy, C _3-8 cycloalkyl;

   m or n are independently selected from 0, 1, 2, 3, 4;

   Y is selected from a 3-14 membered heterocyclyl group directly connected to the quinazoline ring through a C atom on the ring, and the heterocyclyl group is optionally further selected from 1-4 members R ₇ or R ₈ is substituted; R ₇ or R ₈ is independently selected from H, =O, =S, cyano, halogen, nitro, C _1-6 alkyl, -C _0-6 alkylene-OR _a , -C _0-6 alkylene-OC(O)N(R _a ) ₂ , -C _0-6 alkylene-N(R _a ) ₂ , -C _0-6 alkylene-NR _a C( O)R _a , -C _0-6 alkylene-NR _a C(O)N(R _a ) ₂ , -C _0-6 alkylene-NR _a S(O)R _a , -C _0-6 Alkylene-NR _a S(O) ₂ R _a , -C _0-6 alkylene-S(=O)R _a , -C _0-6 alkylene-S(=O) ₂ R _a , - C _0-6 alkylene-SR _a , -C _0-6 alkylene-S(R _a ) ₅ , -C _0-6 alkylene-C(=O)R _a , -C _0-6 alkylene Alkyl-C(=O)OR _a , -C _0-3 alkylene-C(=O)N(R _a ) ₂ , C _2-6 alkenyl, C _2-6 alkynyl, acryloyl, - C _0-6 alkylene-C _3-14 cycloalkyl, -C _0-6 alkylene-(3-14-membered heterocyclic group), -C _0-6 alkylene-C _6-14 aryl Or -C _0-6 alkylene-(5-14-membered heteroaryl), the C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, -C _0-6 alkylene base-C _3-14 cycloalkyl, -C _0-6 alkylene-(3-14-membered heterocyclic group), -C _0-6 alkylene-C _6-14 aryl or -C _0-6 Alkylene-(5-14 membered heteroaryl) may optionally be substituted with one or more R _a ; each R _a is independently selected from H, halogen, hydroxy, amino, oxo, Nitro, cyano, carboxyl, C _1-6 alkyl, C _1-6 hydroxyalkyl, C _1-6 aminoalkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _1-6 Haloalkoxy, C _1-6 heteroalkyl, C _3-8 cycloalkyl, 3-8 membered heterocyclyl, C _6-14 aryl, 5-14 membered heteroaryl, C _1-6 acyl or

   
2. The chiral compound, deuterated compound or pharmaceutically acceptable salt according to claim 1, wherein X is selected from a 4-10-membered heterocyclic group, and the 4-10-membered heterocyclic group is optionally further replaced by 1-4 substituted with a substituent selected from C _1-6 alkyl, cyano, C _1-6 cyanoalkyl, and C _1-6 hydroxyalkyl; preferably

   

   

   

   more preferably

   
3. The chiral compound, deuterated substance or pharmaceutically acceptable salt according to claim 1 or 2, R ¹ is selected from

   

   

   

   preferably

   
4. The chiral compound, deuterated compound or pharmaceutically acceptable salt according to any one of claims 1-3, wherein R ² is selected from H, halogen or C _1-3 alkyl; preferably H.
5. The chiral compound, deuterated compound or pharmaceutically acceptable salt according to any one of claims 1-4, wherein R ³ is selected from H, C _1-6 alkyl, substituted C _1-6 alkyl, C _2-6 alkenyl, substituted C _2-6 alkenyl, C _3-6 cycloalkyl or substituted C _3-6 cycloalkyl; preferably C _1-3 alkyl, C _3-6 cycloalkyl or C _2-4 alkenyl; more preferably ethyl, vinyl or cyclopropyl.
6. The chiral compound, deuterated compound or pharmaceutically acceptable salt according to any one of claims 1-5, wherein R ⁴ or R ⁵ is selected from H, halogen or C _1-6 alkyl; preferably R ⁴ is H and R ⁵ is methyl.
7. The chiral compound, deuterated compound or pharmaceutically acceptable salt according to any one of claims 1-6, wherein R ⁶ is selected from C _1-6 alkoxy, C _3-8 cycloalkyloxy, cyano substituted cyclopropyl C _1-6 alkyleneoxy, said C _1-6 alkoxy optionally further by one or more selected from halogen, hydroxy, methoxy, C _3-8 cycloalkane substituted by the substituent of the group; preferably R ⁶ is selected from

   

   
8. The chiral compound, deuterated substance or pharmaceutically acceptable salt according to any one of claims 1-7, wherein Y is selected from

   

   preferably

   

   

   more preferably

   
9. The chiral compound, deuterated compound or pharmaceutically acceptable salt according to any one of claims 1-8, wherein R ₇ is selected from H, cyano, halogen, C _1-6 alkyl, -C _0-6 Alkylene-OR _a , -C _0-6 alkylene-OC(O)N(R _a ) ₂ , -C _0-6 alkylene-N(R _a ) ₂ , -C _0-6 alkylene base -NR _a C(O)R _a , -C _0-6 alkylene-NR _a C(O)N(R _a ) ₂ , -C _0-6 alkylene-NR _a S(O)R _a , -C _0-6 alkylene-NR _a S(O) ₂ R _a , -C _0-6 alkylene-S(=O)R _a , -C _0-6 alkylene-S(=O ) ₂ R _a , -C _0-6 alkylene-SR _a , -C _0-6 alkylene-S(R _a ) ₅ , -C _0-6 alkylene-C(=O)R _a , -C _0-6 alkylene-C(=O)OR _a , -C _0-3 alkylene-C(=O)N(R _a ) ₂ , acryloyl, -C _0-6 alkylene- C _3-14 cycloalkyl, -C _0-6 alkylene-(3-14-membered heterocyclic group), -C _0-6 alkylene-C _6-14 aryl or -C _0-6 alkylene base-(5-14-membered heteroaryl), the C _1-6 alkyl, -C _0-6 alkylene-C _3-14 cycloalkyl, -C _0-6 alkylene-(3- 14-membered heterocyclyl), -C _0-6 alkylene-C _6-14 aryl or -C _0-6 alkylene-(5-14-membered heteroaryl) optionally can also be replaced by 1 or Replaced by multiple R _a ; each R _a is independently selected from H, halogen, hydroxyl, amino, oxo, cyano, carboxyl, C _1-6 alkyl, C _3-8 cycloalkyl, 3- 8-membered heterocyclyl, C _6-14 -membered aryl, 5-14-membered heteroaryl, C _1-6 acyl or

   

   Preferably R ₇ is selected from H, acryloyl, C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 acyl, -C _0-3 alkylene-C _1-6 alkoxy, C _{1- 6} hydroxyalkyl, C _1-6 cyanoalkyl, -C _0-3 alkylene-S(=O) ₂ -C _1-6 alkyl, C _3-6 keto,

   

   

   -C _0-3 alkylene-C _3-6 cycloalkyl, -C _0-3 alkylene-C _3-8 heterocyclyl, -C _0-3 alkylene-phenyl, -C _{0- 3} alkylene-pyrazolyl, the -C _0-3 alkylene-C _3-6 cycloalkyl, -C _0-3 alkylene-C _3-8 heterocyclyl, -C _0-3 Alkylene-phenyl, -C _0-3 alkylene-pyrazolyl are optionally further selected from one or more of C _1-6 acyl, hydroxy, C _1-6 alkyl, C _3-6 ring Alkyl, halogen,

   

   or cyano substituent, the C _3-8 heterocyclic group contains a heteroatom selected from O or N; more preferably R ₇ is selected from H, C _1-6 alkyl, acetyl,

   

   
10. The chiral compound, deuterated compound or pharmaceutically acceptable salt according to any one of claims 1-9, wherein R ₈ is selected from H or C _1-6 alkyl; preferably H.
11. The chiral compound, deuterated compound or pharmaceutically acceptable salt according to any one of claims 1-10, wherein the chiral compound represented by general formula (I) is selected from compounds of formula (IA)-(ID),

    

    

    ; preferably

    

    wherein the substituents are as defined in claims 1-10.
12. A chiral compound, deuterated substance or pharmaceutically acceptable salt selected from:

    

    

    

    preferably

    

    
13. A pharmaceutical composition, characterized in that the pharmaceutical composition contains the chiral compound, deuterated substance or pharmaceutically acceptable salt according to any one of claims 1-12 and at least one pharmaceutically acceptable adjuvant, and the drug The content of the chiral compound in the composition is higher than that of its corresponding axial chiral isomer; preferably, the content of the chiral compound represented by the general formula (I) in the pharmaceutical composition is higher than that of the active ingredient 51%, 61%, 71%, 81%, 91%, 99% or higher.
14. Application of the chiral compound, deuterated compound, pharmaceutically acceptable salt of any one of claims 1-12 or the pharmaceutical composition of claim 13 in the preparation of medicine; preferably the medicine is an anticancer drug; more preferably the Cancer is mediated by KRAS G12C, HRAS G12C or NRAS G12C mutations.
15. A method of treating and/or preventing cancer, comprising administering a therapeutically effective amount of a chiral compound, deuterated substance, a pharmaceutically acceptable salt or the medicine of any one of claims 1-12 to a treatment object A composition; preferably the cancer is mediated by a KRAS G12C, HRAS G12C or NRAS G12C mutation.

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