CN118359631A

CN118359631A - Polycyclic PI3K alpha inhibitors

Info

Publication number: CN118359631A
Application number: CN202410075812.6A
Authority: CN
Inventors: 刘斌; 陈博
Original assignee: Xuanzhu Biopharmaceutical Co Ltd
Current assignee: Xuanzhu Biopharmaceutical Co Ltd
Priority date: 2023-01-19
Filing date: 2024-01-18
Publication date: 2024-07-19

Abstract

The invention belongs to the technical field of medicines. In particular, the invention relates to a polycyclic PI3K alpha inhibitor compound, a pharmaceutically acceptable salt or a stereoisomer thereof, a pharmaceutical composition and a preparation containing the compound, the pharmaceutically acceptable salt or the stereoisomer thereof, a method for preparing the compound, the pharmaceutically acceptable salt or the stereoisomer thereof, and application of the compound, the pharmaceutically acceptable salt or the stereoisomer thereof in preparing medicines for treating and/or preventing diseases mediated by mutant PI3K alpha and related diseases.

Description

Polycyclic PI3K alpha inhibitors

Technical Field

The invention belongs to the technical field of medicines. In particular, the invention relates to a polycyclic compound which can be used as a mutant PI3K alpha inhibitor, pharmaceutically acceptable salts and stereoisomers thereof, pharmaceutical compositions and preparations containing the compound, the pharmaceutically acceptable salts and stereoisomers thereof, and applications of the compound, the pharmaceutically acceptable salts and stereoisomers thereof.

Background

Cancer is a major cause of death in humans, and global cancer data published by the international agency for research on cancer (IARC) of the world health organization in 2020 indicates that various types of cancer are severely threatening the life health of humans. PI3K signaling pathways can promote cell survival and play an important role in the development and progression of a variety of cancers. Among them, PI3K (phosphatidylinositol-3-kinase) is a key protein on the signaling pathway. In plasma membrane metabolism, PI3K with phosphatidylinositol kinase activity catalyzes PI-4-P (PIP), producing PI-3,4-P ₂(PIP₂), catalyzing PI-4,5-P ₂(PIP₂), producing PI-3,4,5-P ₃(PIP₃). These membrane-bound PIP3 provide anchor sites for a variety of signal transduction proteins, such as AKT (PKB), PDK1, etc., which in turn activate a variety of downstream signaling pathways, producing a broad range of effects that affect cell behavior, such as promoting cell survival, altering cell metabolism.

PI3 ks are classified into type I, type II and type III according to the structure and the specificity of the substrate. Among them, type I PI3K is the most deeply studied and widely studied subtype, and has the most close relation with tumor, and type I PI3K is divided into two subtypes IA and IB. The catalytic subunits of type IA PI3K include p110α, p110β and p110δ, and the catalytic subunits of type IB PI3K are p110γ only, and PI3K can be classified into four subtypes of pi3kα, pi3kβ, pi3kδ and pi3kγ according to the difference of the 4 subunit structures of the catalytic subunits p 110.

As an important bridge linking cell membrane surface receptors and nuclear transcriptional activity, the overactivation of PI3K signaling pathways has been considered one of the most common driving mechanisms in many cancers. The PIK3CA gene (which encodes the p110α subunit) is the most common gene that undergoes point mutations in somatic cells. PIK3CA mutations have a high mutation frequency (e.g., breast cancer: 40%) in some common tumors and occur at three sites of H1047, E542, E545, etc. In tumor cells, PIK3CA mutations enhance kinase activity, continuously stimulate downstream AKT signaling pathways, allow the cell to proliferate independent of growth factors, increase cell invasion and metastasis capacity, and thus, tumors with PIK3CA mutations exist, with a generally poor prognosis. The PIK3CA gene plays an important role in tumorigenesis and development, and is a better target for intervention treatment.

The 2019 PI3K alpha inhibitor Alpelisib was approved for Alpelisib in combination with fulvestrant for use in advanced breast cancer male and postmenopausal female patients with HR positive HER-2 negative, PIK3CA mutations that developed disease progression during or after endocrine therapy. But it has significant adverse effects of hyperglycemia, rash, diarrhea, etc., with incidence of 3 or 4-level hyperglycemia of 33% and 3.9%, respectively. In the Alpelisib clinical phase III trial, patients who were reduced in dose or discontinued in dose due to hyperglycemia reached 54%. There is a great need for safer and more effective agents that inhibit PIK3CA mutations, providing patients with better treatment options.

Disclosure of Invention

The invention aims to provide a compound with a novel structure and good selective inhibition effect on mutant PI3K alpha. Furthermore, the compounds can be used for preparing medicines for treating and/or preventing diseases mediated by mutant PI3K alpha or related diseases.

The technical scheme of the invention is as follows:

In one aspect, the present invention provides a compound of the following formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

Wherein,

When (when)X ₁、X₂ is selected from CH or N, X ₃ is selected from-CR ^1aR^1b-、-N(R^1c) -, -O-, -S-, -C (O) -, -S (O) -or-S (O) ₂ -, when representing a single bond;

When (when) X ₁、X₂ is C, X ₃ is selected from-CR ^1aR^1b-、-N(R^1c) -, -O-, -S-, -C (O) -, -S (O) -or-S (O) ₂ -, when representing a single bond;

When (when) X ₁ is selected from-CH-or-N-, when representing a double bond; x ₂ is-C-, and X ₃ is selected from-CR ^1a -or-N-;

Each X ₄ is independently selected from-CR ^2aR^2b-;

y is selected from-CR ^3aR^3b-、-O-、-NR^3c -or-S-;

L ₁ is selected from-CR ^4aR^4b-CR^4aR^4b-、-CR^4aR^4b-NR^4c -or-CR ^4aR^4b -O-;

Ring a and the X ¹、X² to which it is attached together form a 3-10 membered cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered heteroaryl or 6-10 membered aryl;

Ring B is selected from 3-10 membered cycloalkyl, 3-10 membered heterocycloalkyl, 5-10 membered heteroaryl or 6-10 membered aryl;

Each R ₁, each R ₂ is independently selected from halogen, hydroxy, nitro, cyano, amino, carboxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, amino C _1-6 alkyl 、-NR^aR^b、-OR^a、-SR^a、-NR^a-C(O)-R^b-、-C(O)R^a、-C(O)NR^a、-C(O)OR^a, or the following groups optionally substituted with substituents: -L ₂ -3-10 membered cycloalkyl, -L ₂ -3-10 membered heterocyclyl, -L ₂ -5-10 membered heteroaryl, -L ₂ -6-10 membered aryl; the substituent is selected from halogen, hydroxy, nitro, amino, cyano, carboxyl, C _1-6 alkyl, C _1-6 alkoxy or halogenated C _1-6 alkyl;

Each L ₂ is independently selected from-C _1-6 alkylene-, -C _1-6 alkylene-O-, or-NH-;

R ₃、R₄、R₅ is independently selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, C _1-6 alkyl, haloC _1-6 alkyl, hydroxyC _1-6 alkyl, aminoC _1-6 alkyl, cyanoC _1-6 alkyl, C _1-6 alkoxy, haloC _1-6 alkoxy, hydroxyC _1-6 alkoxy, aminoC _1-6 alkoxy, cyanoC _1-6 alkoxy;

R^1a、R^1b、R^1c、R^2a、R^2b、R^3a、R^3b、R^3c、R^4a、R^4b、R^4c、R^a、R^b Each occurrence is independently selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, C _1-6 alkyl, halo C _1-6 alkyl, hydroxy C _1-6 alkyl, amino C _1-6 alkyl, cyano C _1-6 alkyl, C _1-6 alkoxy, halo C _1-6 alkoxy, hydroxy C _1-6 alkoxy, amino C _1-6 alkoxy, cyano C _1-6 alkoxy;

m, s, t are each independently selected from 0, 1,2, 3 or 4.

In certain embodiments, a compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein,

Ring a and the X ¹、X² to which it is attached together form a 5-10 membered cycloalkyl, 5-10 membered heterocycloalkyl, 5-10 membered heteroaryl or phenyl;

Ring B is selected from phenyl or 5-6 membered heteroaryl;

Each R ₁, each R ₂ is independently selected from halogen, hydroxy, nitro, cyano, amino, carboxy, C _1-4 alkyl, halo C _1-4 alkyl, hydroxy C _1-4 alkyl, amino C _1-4 alkyl 、-NR^aR^b、-OR^a、-SR^a、-NR^a-C(O)-R^b-、-C(O)R^a、-C(O)NR^a、-C(O)OR^a, or the following groups optionally substituted with substituents: -L ₂ -3-6 membered cycloalkyl, -L ₂ -3-6 membered heterocyclyl, -L ₂ -5-6 membered heteroaryl, -L ₂ -phenyl; the substituent is selected from halogen, hydroxy, nitro, amino, cyano, carboxyl, C _1-4 alkyl, C _1-4 alkoxy or halogenated C _1-4 alkyl;

Each L ₂ is independently selected from-C _1-4 alkylene-, -C _1-4 alkylene-O-, or-NH-;

R ₃、R₄、R₅ is independently selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, C _1-4 alkyl, haloC _1-4 alkyl, hydroxyC _1-4 alkyl, aminoC _1-4 alkyl, cyanoC _1-4 alkyl, C _1-4 alkoxy, haloC _1-4 alkoxy, hydroxyC _1-4 alkoxy, aminoC _1-4 alkoxy, cyanoC _1-4 alkoxy;

R^1a、R^1b、R^1c、R^2a、R^2b、R^3a、R^3b、R^3c、R^4a、R^4b、R^4c、R^a、R^b Each occurrence is independently selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, C _1-4 alkyl, halo C _1-4 alkyl, hydroxy C _1-4 alkyl, amino C _1-4 alkyl, cyano C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkoxy, hydroxy C _1-4 alkoxy, amino C _1-4 alkoxy, cyano C _1-4 alkoxy;

m, s, t are each independently selected from 0, 1,2 or 3.

In certain embodiments, ring A and the X ¹、X² to which it is attached together comprise a 5-6 membered monocycloalkyl, 5-6 membered monocycloheteroaryl, phenyl, 8-10 membered fused ring group, 8-10 membered fused heterocyclic group, or 8-10 membered fused heteroaryl group.

In certain embodiments, each R ₁, each R ₂ is independently selected from halogen, hydroxy, nitro, cyano, amino, carboxy, C _1-4 alkyl, halo C _1-4 alkyl, hydroxy C _1-4 alkyl, amino C _1-4 alkyl, -NR ^aR^b、-OR^a、-C(O)OR^a, or the following groups optionally substituted with substituents: -L ₂ -3-6 membered cycloalkyl, -L ₂ -3-6 membered heterocyclyl, -L ₂ -5-6 membered heteroaryl, -L ₂ -phenyl; the substituent is selected from halogen, hydroxy, nitro, amino, cyano, carboxyl, C _1-4 alkyl, C _1-4 alkoxy or halogenated C _1-4 alkyl;

R ^a、R^b is independently at each occurrence selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, C _1-4 alkyl, halo C _1-4 alkyl, C _1-4 alkoxy, or halo C _1-4 alkoxy.

In certain embodiments, L ₂ is selected from -CH₂-、-CH₂-CH₂-、-CH₂-CH₂-CH₂-、-CH₂(CH₃)-CH₂-、-CH₂-O-、-CH₂-CH₂-O-、-O- or-NH-.

,R^1a、R^1b、R^1c、R^2a、R^2b、R^3a、R^3b、R^3c、R^4a、R^4b、R^4c In certain embodiments is independently selected at each occurrence from hydrogen, halogen, C _1-4 alkyl, halogenated C _1-4 alkyl, C _1-4 alkoxy, halogenated C _1-4 alkoxy.

,R^1a、R^1b、R^1c、R^2a、R^2b、R^3a、R^3b、R^3c、R^4a、R^4b、R^4c In certain embodiments is independently hydrogen or C _1-4 alkyl at each occurrence.

In certain embodiments, whenX ₁、X₂ is selected from CH or N, X ₃ is selected from-CH ₂ -, -NH, -O-, -S-, -C (O) -, -S (O) -or-S (O) ₂ -, when representing a single bond;

When (when) X ₁、X₂ is C, X ₃ is selected from-CH ₂ -, -NH, -O-, -S-, -C (O) -, -S (O) -or-S (O) ₂ -, when X represents a single bond;

When (when) X ₁ is selected from-CH-or-N-, when representing a double bond; x ₂ is-C-, X ₃ is selected from-CH-or-N-.

In certain embodiments, whenX ₁、X₂ is independently selected from CH or N, X ₃ is selected from-CH ₂ -, -NH, -O-or-S-when representing a single bond;

When (when) X ₁、X₂ is C, X ₃ is selected from-CH ₂ -, -NH, -O-or-S-, when the bond is represented by the single bond;

In certain embodiments, each X ₄ is independently-CH ₂ -.

In certain embodiments, the compounds of the foregoing general formula (I), pharmaceutically acceptable salts thereof, or stereoisomers thereof, have the structure shown in general formula (II-1),

Wherein X ₁、X₂ is each independently selected from CH or N; r ₁、R₂、R₃、R₄、R₅, Y, ring A, ring B, L ₁、s、t、m、R^1a、R^2a、R^2b are as defined in any of the preceding schemes.

In certain embodiments, the compounds of the foregoing general formula (I), pharmaceutically acceptable salts thereof, or stereoisomers thereof, have the structure shown in general formula (II-2),

Wherein X ₁ is selected from CH or N; x ₂ is C; r ₁、R₂、R₃、R₄、R₅, Y, ring A, ring B, L ₁、s、t、m、R^1a、R^2a、R^2b are as defined in any of the preceding schemes.

In certain embodiments, the compounds of the foregoing general formula (I), pharmaceutically acceptable salts thereof, or stereoisomers thereof, have the structure shown in general formula (II-3),

Wherein each X ₁、X₂ is independently C; r ₁、R₂、R₃、R₄、R₅, Y, ring A, ring B, L ₁、s、t、m、R^1a、R^2a、R^2b are as defined in any of the preceding schemes.

In certain embodiments, the compounds of the foregoing general formula (I), general formula (II-1), general formula (II-2), general formula (II-3), pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein,

Y is selected from-CHR ^3a -, -O-or-NR ^3c -;

l ₁ is selected from-CHR ^4a-CHR^4a-、-CHR^4a-NR^4c -or-CHR ^4a -O-;

ring a and the X ₁、X₂ to which it is attached together form the following group:

When (when) When representing a double bond, each X ₂ is independently C, and each X ₁ is independently selected from N or CH;

When (when) When representing a single bond, each X ₁ and each X ₂ are independently selected from N or CH;

Ring B is selected from phenyl or 6 membered heteroaryl;

Each R ₁, each R ₂ is independently selected from halogen, hydroxy, nitro, cyano, amino, carboxy, C _1-4 alkyl, halo C _1-4 alkyl, hydroxy C _1-4 alkyl, amino C _1-4 alkyl, -NR ^aR^b、-OR^a、-C(O)OR^a;

R ₃、R₄、R₅ is independently selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, C _1-4 alkyl, halogenated C _1-4 alkyl, C _1-4 alkoxy, or halogenated C _1-4 alkoxy;

R ^3a、R^3c、R^4a、R^4c、R^a、R^b is independently at each occurrence selected from hydrogen, halogen, C _1-4 alkyl, halogenated C _1-4 alkyl, C _1-4 alkoxy, or halogenated C _1-4 alkoxy;

m is selected from 0, 1 or 2;

s, t are each independently selected from 0, 1, 2 or 3.

Ring B is selected from phenyl or 6 membered heteroaryl;

m is selected from 0, 1 or 2;

s, t are each independently selected from 0, 1, 2 or 3.

ring B is selected from phenyl, pyridinyl or pyrimidinyl;

Each R ₁, each R ₂ is independently selected from halogen, hydroxy, nitro, cyano, amino, carboxy, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, monofluoromethyl, difluoromethyl, trifluoromethyl, -NR ^aR^b、-OR^a、-C(O)OR^a;

R ₃、R₄、R₅ is independently selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoromethoxy, difluoromethoxy or trifluoromethoxy;

r ^3a、R^3c、R^4a、R^4c、R^a、R^b is independently selected at each occurrence from hydrogen, fluoro, chloro, bromo, iodo, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoromethoxy, difluoromethoxy or trifluoromethoxy;

m is selected from 0, 1 or 2;

s, t are each independently selected from 0, 1, 2 or 3.

In certain embodiments, ring a, and the X ₁、X₂ to which it is attached, together comprise the following group:

In certain embodiments, each R ₁ is independently selected from halogen, carboxyl, C _1-4 alkyl, halogenated C _1-4 alkyl, -OR ^a, OR-C (O) OR ^a;R^a is independently selected from hydrogen, C _1-4 alkyl, OR halogenated C _1-4 alkyl.

In certain embodiments, each R ₁ is independently selected from halogen, carboxyl, C _1-4 alkyl, halogenated C _1-4 alkyl; r ^a is independently selected from hydrogen or C _1-4 alkyl.

In certain embodiments, each R ₁ is independently selected from fluorine, chlorine, carboxyl, methyl, ethyl, monofluoromethyl, difluoromethyl, or trifluoromethyl.

In certain embodiments, each R ₂ is independently selected from halogen, C _1-4 alkyl, halogenated C _1-4 alkyl, OR —or ^a;R^a is independently selected from hydrogen, C _1-4 alkyl, OR halogenated C _1-4 alkyl.

In certain embodiments, each R ₂ is independently selected from fluorine, chlorine, methyl, ethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, OR-OR ^a;R^a is independently selected from hydrogen, methyl, ethyl, monofluoromethyl, difluoromethyl, OR trifluoromethyl.

In certain embodiments, R ₃、R₄、R₅ is each independently hydrogen, halogen, C _1-4 alkyl, or halogenated C _1-4 alkyl.

In certain embodiments, each R ₃、R₅ is independently hydrogen.

In certain embodiments, R ₄ is selected from hydrogen, fluoro, chloro, methyl, monofluoromethyl, difluoromethyl, or trifluoromethyl.

In some embodiments of the present invention, in some embodiments, Y is selected from-CH ₂ -, -O-or-NH-.

In certain embodiments, Y is O.

In certain embodiments, L ₁ is selected from-CHR ^4a-CHR^4a-、-CHR^4a-NR^4c -or-CHR ^4a-O-;R^4a、R^4c, each independently selected from hydrogen or C _1-6 alkyl.

In certain embodiments, L ¹ is-CHR ^4a-NR^4c-;R^4a、R^4c each independently selected from hydrogen or C _1-4 alkyl.

In certain embodiments, m is selected from 0 or 1.

In certain embodiments, s is selected from 1 or 2.

In certain embodiments, s is 1 and R ₁ is carboxyl.

In certain embodiments, t is selected from 0, 1, or 2.

In certain embodiments, the compounds of the foregoing general formula (I), general formula (II-1), general formula (II-2), general formula (II-3), pharmaceutically acceptable salts thereof, or stereoisomers thereof, have structures as shown in general formula (III-1), general formula (III-2), general formula (III-3),

Wherein Y' is each independently selected from CH or N;

X ₁、X₂、R₁、R₂、R₃、R₄、R₅, Y, and ring A, s, t, m, R ^1a、R^2a、R^2b、R^4a、R^4c are as defined in any of the preceding schemes.

In certain embodiments, the compounds described by the foregoing general formula (I), general formula (II-1), general formula (II-2), general formula (II-3), general formula (III-1), general formula (III-2), general formula (III-3), pharmaceutically acceptable salts thereof, and stereoisomers thereof, are selected from the following compounds:

The invention also provides a pharmaceutical composition, which contains the compounds of the general formula (I), the general formula (II-1), the general formula (II-2), the general formula (II-3), the general formula (III-1), the general formula (III-2) and the general formula (III-3), pharmaceutically acceptable salts or stereoisomers thereof, and one or more medicinal carriers and/or diluents; the pharmaceutical composition can be prepared into any clinically or pharmaceutically acceptable dosage form, such as tablets, capsules, pills, granules, solutions, suspensions, syrups, injections (including injections, sterile powders for injection and concentrated solutions for injection), suppositories, inhalants or sprays and the like.

In certain embodiments of the present invention, the above-described pharmaceutical formulations may be administered orally, parenterally, rectally, or pulmonary, etc., to a patient or subject in need of such treatment. For oral administration, the pharmaceutical composition may be formulated into oral preparations, for example, into conventional oral solid preparations such as tablets, capsules, pills, granules, etc.; can also be made into oral liquid preparation such as oral solution, oral suspension, syrup, etc. When the composition is formulated into oral preparations, suitable fillers, binders, disintegrants, lubricants, etc. may be added. For parenteral administration, the pharmaceutical preparations may also be formulated as injections, including injectable solutions, injectable sterile powders and injectable concentrated solutions. When the injection is prepared, the conventional method in the existing pharmaceutical field can be adopted for production, and when the injection is prepared, no additive can be added, and the proper additive can be added according to the property of the medicine. For rectal administration, the pharmaceutical composition may be formulated as suppositories and the like. For pulmonary administration, the pharmaceutical composition may be formulated as an inhalant or spray, etc.

The pharmaceutically acceptable carrier and/or diluent useful in the pharmaceutical composition or pharmaceutical formulation of the present invention may be any conventional carrier and/or diluent in the pharmaceutical formulation arts, and the choice of the particular carrier and/or diluent will depend on the mode of administration or type and state of disease for the particular patient being treated. The preparation of suitable pharmaceutical compositions for specific modes of administration is well within the knowledge of those skilled in the pharmaceutical arts.

In a further aspect, the invention also relates to the use of a compound of the aforementioned general formula (I), general formula (II-1), general formula (II-2), general formula (II-3), general formula (III-1), general formula (III-2), general formula (III-3), a pharmaceutically acceptable salt thereof or a stereoisomer thereof for the preparation of a medicament for the prophylaxis and/or treatment of diseases and related disorders mediated by mutant PI3K alpha, which medicament can be used in combination with one or more other medicaments for the prophylaxis or treatment of diseases and related disorders mediated by mutant PI3K alpha. The disease and related conditions are selected from cancers, including carcinoma in situ and metastatic cancers, or benign tumors. Further, the cancers include, but are not limited to, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, thyroid cancer, female genital tract cancer, lymphoma, neurofibromas, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, small cell lung cancer, non-small cell lung cancer, gastrointestinal stromal tumor, mast cell tumor, multiple myeloma, melanoma, leukemia, glioma or sarcoma, and the like.

In certain embodiments, the mutant pi3kα has the E542K, E K and H1047R mutations.

In certain embodiments, the mutant pi3kα has an H1047R mutation.

Furthermore, the invention also relates to the use of the pharmaceutical preparations of the compounds of the general formula (I), the general formula (II-1), the general formula (II-2), the general formula (II-3), the general formula (III-1), the general formula (III-2) and the general formula (III-3), pharmaceutically acceptable salts or stereoisomers thereof for preparing medicaments, wherein the medicaments can be combined with one or more medicaments to treat and/or prevent diseases and related diseases mediated by mutant PI3K alpha.

In another aspect, the application relates to the aforementioned general formula (I), general formula (II-1), general formula (II-2), general formula (II-3), general formula (III-1), general formula (III-2), general formula (III-3), the compounds described in formula (III-3), their pharmaceutically acceptable salts or the stereoisomers thereof, may be administered alone or in combination with one or more second therapeutically active agents for use in combination with the PI3K alpha inhibitor compounds of the application in the treatment and/or prevention of diseases and related disorders mediated by mutant PI3K alpha. Thus, in certain embodiments, the pharmaceutical composition further comprises one or more second therapeutically active agents. Optionally, the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers and/or diluents.

In certain embodiments, the individual components to be combined (e.g., a compound of the invention, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, and a second therapeutically active agent) can be administered simultaneously or sequentially and separately administered in sequence. For example, the second therapeutically active agent may be administered before, simultaneously with, or after administration of the compound of the invention, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. Furthermore, the individual components to be combined can also be administered jointly in the form of the same formulation or in the form of separate different formulations.

In another aspect, the present invention also provides a method of treating diseases and related conditions mediated by mutant pi3kα, comprising administering to a patient in need thereof an effective amount of a compound of formula (I), a pharmaceutically acceptable salt or stereoisomer thereof, a formulation or a pharmaceutical composition as described above; the diseases and related conditions mediated by mutant pi3kα are as defined above.

By "effective amount" is meant an amount of a drug capable of alleviating, delaying, inhibiting or curing a condition in a subject. The size of the dose administered is determined by the mode of administration of the drug, the pharmacokinetics of the agent, the severity of the disease, the individual sign (sex, weight, height, age) of the subject, etc.

[ Definitions and general terms ]

In the description and claims of the present application, compounds are named according to chemical structural formulas, and if the same compounds are represented, the named and chemical structural formulas of the compounds are not identical, the chemical structural formulas are used as references.

In the present application, unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by one of ordinary skill in the art, however, for a better understanding of the present application, the following definitions of some terms are provided. When the definition and interpretation of terms provided by the present application are not identical to the meanings commonly understood by those skilled in the art, the definition and interpretation of terms provided by the present application is in control.

"Halogen" as used herein refers to a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

"C _1-6 alkyl" as used herein means straight or branched chain alkyl groups containing 1 to 6 carbon atoms, including for example "C _1-4 alkyl", "C _1-3 alkyl", "C _1-2 alkyl", "C _2-6 alkyl", "C _2-5 alkyl", "C _2-4 alkyl", "C _2-3 alkyl", "C _3-6 alkyl", "C _3-5 alkyl", "C _3-4 alkyl" and the like, specific examples include, but are not limited to: methyl, ethyl, n-propyl (propyl), isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 1, 2-dimethylpropyl, and the like. As used herein, "C _1-4 alkyl" refers to a specific example of C _1-6 alkyl containing 1 to 4 carbon atoms.

As used herein, "C _1-6 alkoxy" refers to "C _1-6 alkyl-O-", and "C _1-6 alkyl" is as defined above. As used herein, "C _1-4 alkoxy" refers to "C _1-4 alkyl-O-", and "C _1-4 alkyl" is as defined above.

As used herein, "hydroxy C _1-6 alkyl, amino C _1-6 alkyl, halo C _1-6 alkyl, carboxy C _1-6 alkyl" means that one or more hydrogens of the C _1-6 alkyl group are replaced with one or more hydroxy, amino, halogen, or carboxy groups, respectively. The "C _1-6 alkyl" is as defined above.

The term "hydroxy C _1-6 alkoxy, amino C _1-6 alkoxy, halo C _1-6 alkoxy" as used herein means that one or more hydrogens of the "C _1-6 alkoxy" are replaced with one or more hydroxy, amino or halogen groups.

The terms "C _1-6 alkylamino, C _1-6 alkoxyamino, C _1-6 alkylcarbonyl, C _1-6 alkoxycarbonyl, C _1-6 alkylthio carbonyl, C _1-6 alkylsulfonyl, C _1-6 alkylamido, di (C _1-6 alkyl) amino, di (C _1-6 alkyl) aminocarbonyl" as used herein refer to C _1-6 alkyl-NH-, C _1-6 alkyl-O-NH-, C _1-6 alkyl-C (O) -, C _1-6 alkoxy-C (O) -, C _1-6 alkylthio-C (O) -, C _1-6 alkyl-S (O) ₂-、C_1-6 alkyl-C (O) -NH-,

The "6-10 membered aryl" described herein includes "6-8 membered monocyclic aryl" and "8-10 membered condensed ring aryl".

"6-8 Membered monocyclic aryl" as used herein refers to monocyclic aryl groups containing 6-8 ring carbon atoms, examples of which include, but are not limited to: phenyl, cyclooctatetraenyl, and the like; phenyl is preferred.

The term "8-to 10-membered condensed ring aryl" as used herein refers to an unsaturated, aromatic cyclic group containing 8 to 10 ring carbon atoms, preferably "9-to 10-membered condensed ring aryl", which is formed by sharing two or more adjacent atoms with each other by two or more cyclic structures, and specific examples thereof are naphthyl and the like.

The "5-10 membered heteroaryl" as used herein includes "5-8 membered monocyclic heteroaryl" and "8-10 membered fused heteroaryl".

"5-8 Membered monocyclic heteroaryl" as used herein refers to a monocyclic cyclic group having aromaticity which contains 5-8 ring atoms, at least one of which is a heteroatom, such as a nitrogen atom, an oxygen atom or a sulfur atom. Optionally, a ring atom (e.g., a carbon atom, a nitrogen atom, or a sulfur atom) in the cyclic structure may be oxo. "5-8 membered monocyclic heteroaryl" includes, for example, "5-7 membered monocyclic heteroaryl", "5-6 membered nitrogen containing monocyclic heteroaryl", "6 membered nitrogen containing monocyclic heteroaryl", etc., wherein the heteroatoms in the "nitrogen containing heteroaryl" contain at least one nitrogen atom, for example, contain only 1 or 2 nitrogen atoms, or contain one nitrogen atom and 1 or 2 other heteroatoms (e.g., oxygen and/or sulfur atoms), or contain 2 nitrogen atoms and 1 or 2 other heteroatoms (e.g., oxygen and/or sulfur atoms). Specific examples of the "5-8 membered monocyclic heteroaryl" include, but are not limited to, furyl, thienyl, pyrrolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, 1,2, 3-triazolyl, 1,2, 4-triazolyl, 1,2, 3-oxadiazolyl, 1,2, 4-oxadiazolyl, 1,2, 5-oxadiazolyl, 1,3, 4-oxadiazolyl, pyridyl, 2-pyridonyl, 4-pyridonyl, pyrimidinyl, pyridazinyl, pyrazinyl, 1,2, 3-triazinyl, 1,3, 5-triazinyl, 1,2,4, 5-tetrazinyl, azepanyl, 1, 3-diazinoheptenyl, azocyclotetraenyl and the like. The "5-6 membered heteroaryl" refers to a specific example in which 5-8 membered heteroaryl contains 5-6 ring atoms.

The "8-to 10-membered fused heteroaryl group" as used herein refers to an unsaturated aromatic ring structure containing 8 to 10 ring atoms (at least one of which is a heteroatom such as a nitrogen atom, an oxygen atom or a sulfur atom) formed by two or more ring structures sharing two adjacent atoms with each other. Optionally, a ring atom (e.g., a carbon atom, a nitrogen atom, or a sulfur atom) in the cyclic structure may be oxo. Including "9-10 membered fused heteroaryl", "8-9 membered fused heteroaryl", "9-10 membered fused heteroaryl containing 1-2 heteroatoms selected from nitrogen, oxygen or sulfur" and the like, which may be fused in such a manner as to be benzo 5-6 membered heteroaryl, 5-6 membered heteroaryl and the like; specific examples include, but are not limited to: pyrrolopyrroles, pyrrolofurans, pyrazolopyrroles, pyrazolothiophenes, furanthiophenes, pyrazolooxazoles, benzofuranyl, benzisofuranyl, benzothienyl, indolyl, isoindolyl, benzoxazolyl, benzimidazolyl, indazolyl, benzotriazolyl, quinolinyl, 2-quinolinonyl, 4-quinolinonyl, 1-isoquinolonyl, isoquinolinyl, acridinyl, phenanthridinyl, benzopyridazinyl, phthalazinyl, quinazolinyl, quinoxalinyl, purinyl, naphthyridinyl, and the like.

The "3-10 membered cycloalkyl" as used herein includes "3-7 membered monocyclic cycloalkyl" and "8-10 membered fused ring cycloalkyl".

"3-7 Membered monocyclic cycloalkyl" as used herein refers to a saturated or partially saturated and non-aromatic monocyclic ring group containing 3-7 ring atoms, including "3-6 membered monocyclic cycloalkyl", "5-6 membered monocyclic cycloalkyl", specific examples include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexene, or the like.

"8-10 Membered fused ring cycloalkyl" as used herein refers to a saturated or partially saturated, non-aromatic cyclic group containing 8-10 ring atoms formed by two or more cyclic structures sharing two adjacent atoms with each other, examples of which include, but are not limited to: Etc.

As used herein, "3-8 membered cycloalkyl" refers to a specific example of "3-10 membered cycloalkyl" containing 3-8 ring carbon atoms.

The "3-10 membered heterocyclic group" as used herein includes "3-7 membered single heterocyclic group" and "8-10 membered condensed heterocyclic group".

"3-7 Membered heterocyclic group" as used herein refers to a saturated or partially saturated and non-aromatic monocyclic ring group containing at least one heteroatom (e.g., containing 1,2, 3,4 or 5) and having 3 to 7 ring atoms, the heteroatom being a nitrogen atom, an oxygen atom and/or a sulfur atom, optionally, a ring atom in the ring structure (e.g., carbon atom, nitrogen atom or sulfur atom) may be oxo. The "3-7 membered monocyclic group" described in the present invention includes "3-7 membered saturated monocyclic group" and "3-7 membered partially saturated monocyclic group". Preferably, the "3-7 membered mono-heterocyclic group" according to the present invention contains 1-3 heteroatoms; preferably, the "3-7 membered mono-heterocyclic group" according to the present invention contains 1-2 hetero atoms, and the hetero atoms are selected from nitrogen atoms and/or oxygen atoms; preferably, the "3-7 membered mono-heterocyclic group" according to the present invention contains 1 nitrogen atom. The "3-7 membered mono-heterocyclic group" is preferably "3-6 membered mono-heterocyclic group", "4-7 membered mono-heterocyclic group", "4-6 membered mono-heterocyclic group", "6-8 membered mono-heterocyclic group", "5-7 membered mono-heterocyclic group", "5-6 membered mono-heterocyclic group", "3-6 membered saturated mono-heterocyclic group", "5-6 membered saturated mono-heterocyclic group", "3-6 membered nitrogen-containing mono-heterocyclic group", "3-6 membered saturated nitrogen-containing mono-heterocyclic group", "5-6 membered saturated nitrogen-containing mono-heterocyclic group" or the like. For example, containing only 1 or2 nitrogen atoms, or containing one nitrogen atom and1 or2 other heteroatoms (e.g., oxygen and/or sulfur atoms). Specific examples of "3-7 membered mono-heterocyclyl" include, but are not limited to: aziridinyl, 2H-aziridinyl, diazinoalkyl, 3H-diazinopropenyl, azetidinyl, 1, 4-dioxanyl, 1, 3-dioxanyl, 1, 4-dioxanyl, tetrahydrofuranyl, dihydropyrrolyl, pyrrolidinyl, imidazolidinyl, 4, 5-dihydroimidazolyl, pyrazolidinyl, 4, 5-dihydropyrazolyl, 2, 5-dihydrothienyl, tetrahydrothienyl, 4, 5-dihydrothiazolyl, thiazolidinyl, piperidinyl, tetrahydropyridinyl, piperidonyl, tetrahydropyridinonyl, dihydropyridinonyl, piperazinyl, morpholinyl, 4, 5-dihydro-oxazolyl, 4, 5-dihydro-isoxazolyl, 2, 3-dihydro-isoxazolyl, oxazolyl, 2H-1, 2-oxazinyl, 6H-1, 2-oxazinyl, 4H-1, 3-oxazinyl, 6H-1, 3-oxazinyl, 3H-1, 2-oxazinyl, 4H-oxazinyl, 4, 3H-oxazinyl, 4H-1, 2-oxazinyl, 3H-oxazinyl, 4, 3-oxazinyl, 4H-1, 3-oxazinyl, 3-H-oxazinyl, 4-H-1, 3-H-oxazinyl, 3-H-2-oxazinyl, etc.

The "8-10 membered fused heterocyclic group" as used herein refers to a saturated or partially saturated, non-aromatic cyclic group containing 8 to 10 ring atoms and at least one ring atom being a heteroatom, formed by two or more cyclic structures sharing two adjacent atoms with each other, wherein one of the rings may be an aromatic ring, but the whole of the fused ring is not aromatic, and the heteroatom is a nitrogen atom, an oxygen atom and/or a sulfur atom, optionally, a ring atom (e.g., a carbon atom, a nitrogen atom or a sulfur atom) in the cyclic structure may be oxo, including, but not limited to, "8-9 membered fused heterocyclic group", "9-10 membered fused heterocyclic group", and the like; specific examples of the "8-to 10-membered fused heterocyclic group" include, but are not limited to: pyrrolidinyl-cyclopropyl, cyclopentylazacyclopropyl, pyrrolidinyl-cyclobutyl, pyrrolidinyl-piperidyl, pyrrolidinyl-piperazinyl, pyrrolidinyl-morpholinyl, piperidinyl-morpholinyl, benzopyrrolidinyl, benzocyclopentyl, benzocyclohexyl, benzotetrahydrofuranyl, benzopyrrolidinyl, pyrimidotetrahydropyranyl; tetrahydroimidazo [4,5-c ] pyridinyl, 3, 4-dihydroquinazolinyl, 1, 2-dihydroquinoxalinyl, benzo [ d ] [1,3] dioxolyl, 2H-chromen-2-onyl, 4H-chromen-yl, 4H-chromen-4-onyl, 4H-1, 3-benzoxazolyl, 4, 6-dihydro-1H-furo [3,4-d ] imidazolyl, 3a,4,6 a-tetrahydro-1H-furo [3,4-d ] imidazolyl, 4, 6-dihydro-1H-thieno [3,4-d ] imidazolyl, 4, 6-dihydro-1H-pyrrolo [3,4-d ] imidazolyl, octahydro-benzo [ d ] imidazolyl, decahydroquinolinyl, hexahydrothienoimidazoyl, hexahydrofuroimidazoyl, 4,5,6, 7-tetrahydro-1H-benzo [3,4-d ] imidazolyl, octahydro-pyrrolo [3,4-d ] imidazolyl, and the like.

The term "carbon atom, nitrogen atom or sulfur atom is oxo" as used herein refers to a structure that forms c= O, N = O, S =o or SO ₂.

"Optionally substituted" as used herein refers to both cases where one or more atoms on the substituted group may be "substituted" or "unsubstituted" with one or more substituents.

The term "pharmaceutically acceptable salts" as used herein refers to salts of acidic functional groups present in the compounds (e.g., -COOH, -OH, -SO ₃ H, etc.) with suitable inorganic or organic cations (bases), including salts with alkali metals or alkaline earth metals, ammonium salts, and salts with nitrogen-containing organic bases; and salts of basic functional groups (e.g., -NH ₂, etc.) present in the compounds with suitable inorganic or organic anions (acids), including salts with inorganic or organic acids (e.g., carboxylic acids, etc.).

"Stereoisomers" as used herein refers to compounds of the invention which contain one or more asymmetric centers and are thus useful as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. The compounds of the present invention may have asymmetric centers that each independently produce two optical isomers. The scope of the present invention includes all possible optical isomers and mixtures thereof. The compounds of the present invention, if they contain olefinic double bonds, include cis-isomers and trans-isomers unless specified otherwise. The compounds described herein may exist in tautomeric (one of the functional group isomers) forms having different points of attachment of hydrogen through displacement of one or more double bonds, for example, the keto and enol forms thereof are keto-enol tautomers. Each tautomer and mixtures thereof are included within the scope of the present invention. Enantiomers, diastereomers, racemates, meso, cis-trans isomers, tautomers, geometric isomers, epimers, mixtures thereof and the like of all compounds are included within the scope of the present invention.

The term "dosage form" as used herein refers to a form of a medicament formulated for clinical use, including, but not limited to, powders, tablets, granules, capsules, solutions, emulsions, suspensions, injections (including injectable solutions, injectable sterile powders and injectable concentrated solutions), sprays, aerosols, powder sprays, lotions, liniments, ointments, plasters, pastes, patches, gargles or suppositories, more preferably powders, tablets, granules, capsules, solutions, injections, ointments, gargles or suppositories.

Advantageous effects of the invention

(1) The compound, the pharmaceutically acceptable salt or the stereoisomer thereof has excellent inhibitory activity on the mutant PI3K alpha, and can treat diseases mediated by the mutant PI3K alpha and related diseases;

(2) The compound, the pharmaceutically acceptable salt or the stereoisomer thereof has good pharmacokinetic properties, longer action and high bioavailability;

(3) The compound, the pharmaceutically acceptable salt or the stereoisomer thereof has good safety;

(4) The compound has the advantages of simple preparation process, high purity of the medicine, stable quality and easy mass industrial production.

Detailed Description

The present invention will be described in further detail with reference to the following examples. It should not be construed that the scope of the invention is limited to the following examples. All techniques that can be realized based on the above-described aspects of the present invention are within the scope of the present invention.

Preparation example of the Compounds of the invention

DCM: dichloromethane THF: tetrahydrofuran NaHMDS: sodium bis (trimethylsilyl) amide

EA: ethyl acetate TEA: triethylamine DMF: n, N-dimethylformamide

PE: petroleum ether TFA: trifluoroacetic acid DIPEA: n, N-diisopropylethylamine Pd (PPh ₃)₂Cl₂: bis (triphenylphosphine) palladium dichloride JohnPhos: bis (tert-butylphosphine) biphenyl Xantphos:4, 5-bis (triphenylphosphine) -9, 9-dimethylxanthene LiHMDS: lithium bis (trimethylsilyl) amide HATU:2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate

CMMTPP: methoxymethylene triphenylphosphine

Pd (dppf) Cl ₂: 1,1' -bis-diphenylphosphino ferrocene palladium dichloride

Preparation example 1: preparation of 2- ((1- (9-methyl-7-oxo-4 b,5,6, 14-tetrahydro-7H-chromene [3',2':5,6] pyrido [2,1-a ] isoindol-11-yl) ethyl) amino) benzoic acid (Compound 1)

1.1 Preparation of 1- (2-iodobenzyl) piperidine-2, 6-dione (Int-3)

DMF (200 mL), piperidine-2, 6-dione (Int-1) (20.0 g,176.8 mmol), 1- (bromomethyl) -2-iodobenzene (Int-2) (52.6 g,176.8 mmol) and cesium carbonate (57.6 g,176.8 mmol) were added sequentially to a 1L reaction flask and reacted at 60℃for 2 hours. Cooling to 20deg.C, extracting with water and EA, concentrating the organic phase under reduced pressure, and purifying the crude product with silica gel column (EA: PE=1:3) to obtain the target compound (20.0 g, yield 34.4%).

Preparation of 10 b-hydroxy-2, 3,6,10 b-tetrahydropyrido [2,1-a ] isoindol-4 (1H) -one (Int-4)

DMF (200 mL), int-3 (20.0 g,60.8 mmol) and cesium fluoride (11.1 g,73.0 mmol) were added sequentially to a 1L reaction flask, and tributyltin trimethylsilicate (26.5 g,73.0 mmol) was added dropwise at 0deg.C under nitrogen protection and reacted for 2 hours at 0deg.C. The reaction was quenched with water and DCM and the organic phase was purified by column on silica gel (EA: pe=1:3) to give the title compound (7.0 g, 56.7% yield).

Preparation of 2,3,6,10 b-tetrahydropyrido [2,1-a ] isoindol-4 (1H) -one (Int-5)

DCM (200 mL), int-4 (10.5 g,51.7 mmol), p-toluenesulfonic acid (1.8 g,10.3 mmol) and palladium on carbon (10.5 g,10% wt) were added sequentially to a 500mL reaction flask, and the mixture was reacted at 20℃for 16 hours under hydrogen protection at 20 atm. Filtration and purification of the filtrate over a silica gel column (EA: pe=1:1) gave the title compound (5.0 g, yield 51.7%).

4.Preparation of methyl 3-bromo-2-hydroxy-5-methylbenzoate (Int-7)

Chloroform (100 mL) and methyl 2-hydroxy-5-methylbenzoate (Int-6) (9.0 g,54.2 mmol) were sequentially added to a 500mL reaction flask, and bromine (8.7 g,54.2 mmol) was added at 0deg.C to react for 2 hours at 20deg.C. Aqueous sodium bicarbonate was added to extract, and the organic phase was concentrated to dryness under reduced pressure to give the objective compound (9.0 g, yield 67.8%).

Preparation of methyl 3-bromo-2- (methoxymethoxy) -5-methylbenzoate (Int-8)

DMF (100 mL) and Int-7 (9.0 g,36.7 mmol) were added sequentially to a 1L reaction flask, sodium hydride (1.8 g,44.0mmol,60% wt) was added at 0deg.C, and the mixture was reacted at 0deg.C for 0.5 hours, chloromethyl methyl ether (3.5 g,44.0 mmol) was added, and the mixture was reacted at 20deg.C for 2 hours. Water and EA were added for extraction, and the organic phase was concentrated under reduced pressure, and the crude product was purified by silica gel column (EA: pe=1:10) to give the objective compound (9.0 g, yield 84.9%).

Preparation of 3- (3-bromo-2- (methoxymethoxy) -5-methylbenzoyl) -2,3,6,10 b-tetrahydropyrido [2,1-a ] isoindol-4 (1H) -one (Int-9)

THF (30 mL) and Int-5 (3.0 g,16.0 mmol) were sequentially added to a 500mL reaction flask, a tetrahydrofuran solution of LiHMDS (32 mL,32.0mmol,1 mol/L) was added dropwise at 70℃and reacted at 70℃for 1 hour, and Int-8 (4.6 g,16.0 mmol) was added and reacted at 20℃for 2 hours. Saturated aqueous ammonium chloride and EA were added for extraction, the reaction system was concentrated under reduced pressure, and purified by silica gel column (EA: pe=1:4) to give the objective compound (2.5 g, yield 35.1%).

7.3 Preparation of 3- (3-bromo-2-hydroxy-5-methylbenzoyl) -2,3,6,10 b-tetrahydropyrido [2,1-a ] isoindol-4 (1H) -one (Int-10)

DCM (10 mL), int-9 (2.5 g,5.6 mmol) and TFA (5 mL) were added sequentially to a 50mL reaction flask and reacted at 20℃for 2 hours. Concentrating under reduced pressure, extracting with DCM and sodium bicarbonate aqueous solution, concentrating the organic phase under reduced pressure, pulping with n-heptane, and filtering. The title compound (1.3 g, yield 57.8%) was obtained.

8.11 Preparation of bromo-9-methyl-4 b,5,6, 14-tetrahydro-7H-chromene [3',2':5,6] pyrido [2,1-a ] isoindol-7-one (Int-11)

To a 100mL reaction flask were successively added dichloroethane (20 mL), int-10 (1.3 g,3.2 mmol), trifluoromethanesulfonic anhydride (1.4 g,4.8 mmol) and DIPEA (0.6 g,4.8 mmol), and reacted at 20℃for 2 hours. Aqueous sodium bicarbonate and EA were added to extract, the organic phase was concentrated to dryness under reduced pressure, and EA was added to slurry and filtered. The title compound (400 mg, yield 32.2%) was obtained.

9.Preparation of 11-acetyl-9-methyl-4 b,5,6, 14-tetrahydro-7H-chromene [3',2':5,6] pyrido [2,1-a ] isoindol-7-one (Int-12)

A50 mL reaction flask was charged with 1, 4-dioxane (10 mL), int-11 (400 mg,1.05 mmol), tributyl (1-ethoxyethylene) tin (Int-37) (45 mg,1.26 mmol), palladium acetate (47 mg,0.21 mmol), and Xantphos (185 mg,0.32 mmol) in this order, and reacted at 100℃for 2 hours. Dilute hydrochloric acid (1 mol/L,1.1 mL) was added at 50 ℃, reacted at 50 ℃ for 0.5 hours, extracted with water and EA at 20 ℃, and the organic phase was concentrated under reduced pressure to the remaining 5mL and filtered. The title compound (240 mg, yield 66.5%) was obtained.

Preparation of 11- (1-hydroxyethyl) -9-methyl-4 b,5,6, 14-tetrahydro-7H-chromeno [3',2':5,6] pyrido [2,1-a ] isoindol-7-one (Int-13)

DCM (5 mL), methanol (5 mL) and Int-12 (240 mg,0.69 mmol) were added sequentially to a 100mL reaction flask, sodium borohydride (52 mg,1.38 mmol) was added at 0deg.C, and the reaction was carried out at 20deg.C for 0.5 hours. Water and DCM were added and extracted, the organic phase concentrated to the remaining 1mL under reduced pressure and filtered. The title compound (100 mg, yield 41.5%) was obtained.

11.11 Preparation of (1-bromoethyl) -9-methyl-4 b,5,6, 14-tetrahydro-7H-chromene [3',2':5,6] pyrido [2,1-a ] isoindol-7-one (Int-14)

DCM (5 mL) and Int-13 (100 mg,0.29 mmol) were added sequentially to a 50mL reaction flask, and boron tribromide (236 mg,0.87 mmol) was added at 0deg.C and reacted for 1 hour at 30deg.C. Quench with water at 0 ℃, adjust ph=8 with aqueous sodium bicarbonate, extract with dcm and concentrate the organic phase under reduced pressure. The title compound (56 mg, yield 47.5%) was obtained.

Preparation of methyl 2- ((1- (9-methyl-7-oxo-4 b,5,6, 14-tetrahydro-7H-chromene [3',2':5,6] pyrido [2,1-a ] isoindol-11-yl) ethyl) amino) benzoate (Int-16)

DMF (5 mL), int-14 (56 mg,0.14 mmol), methyl 2-aminobenzoate (Int-15) (42 mg,0.28 mmol) and DIPEA (36 mg,0.28 mmol) were added sequentially to a 50mL reaction flask and reacted at 80℃for 16 hours. Water and DCM were added for extraction, and the organic phase was concentrated under reduced pressure and the crude product purified by silica gel column (EA: pe=1:1) to give the title compound (35 mg, yield 53.3%).

Preparation of- ((1- (9-methyl-7-oxo-4 b,5,6, 14-tetrahydro-7H-chromene [3',2':5,6] pyrido [2,1-a ] isoindol-11-yl) ethyl) amino) benzoic acid (Compound 1)

To a 50mL reaction flask were successively added methanol (2 mL), water (2 mL), int-16 (35 mg,0.073 mmol) and lithium hydroxide monohydrate (16 mg,0.37 mmol), and reacted at 50℃for 2 hours. Dilute aqueous hydrochloric acid was added to adjust ph=3, ea was extracted, the organic phase was concentrated under reduced pressure, THF was added to slurry, and filtration was performed to give the objective compound (10 mg, yield 29.4%).

Molecular formula C ₂₉H₂₆N₂O₄ molecular weight 466.54LC-MS (M/e): 467.14 (M+H ⁺)

¹H-NMR(400MHz,DMSO-d6)δ:8.60(br,1H),7.65～7.81(m,2H),7.23～7.55(m,6H),6.53(s,2H),4.95～5.31(m,4H),3.60～3.80(m,1H),2.69～2.89(m,1H),2.50～2.67(m,2H),2.31(s,3H),1.66(s,3H).

Preparation example 2: preparation of 2- ((1- (9-methyl-7-oxo-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen [3,2-c ] quinolin-11-yl) ethyl) amino) benzoic acid (Compound 2)

1.2 Preparation of tert-butyl 2- (2- ((methylsulfonyl) oxy) ethyl) piperidine-1-carboxylate (Int-18)

To a solution of tert-butyl 2- (2-hydroxyethyl) piperidine-1-carboxylate (Int-17) (10.0 g,43.6 mmol) in DCM (100 mL) was added TEA (8.8 g,87.0 mmol) and methylsulfonyl chloride (7.5 g,65.4 mmol) dropwise at 0deg.C for 30 min. The system was quenched with water, the organic phase extracted with DCM, concentrated and spun-dried to give crude product which was used directly in the next step.

2.Preparation of diethyl 2- (2- (1- (tert-Butoxycarbonyl) piperidin-2-yl) ethyl) malonate (Int-19)

To a solution of diethyl malonate (13.1 g,87.2 mmol) in THF (100 mL) at 0 ℃ was added sodium hydride (60%) (3.5 g,87.5 mmol), and reacted for 30 minutes, then Int-18 (crude product of the above step) was added, and the mixture was allowed to react at 55 ℃ for 24 hours. The pH was adjusted to 4 with 1M dilute hydrochloric acid at 0deg.C, the organic phase was extracted with EA, dried and concentrated, and purified by silica gel column (EA: PE=1:5) to give 8.0g of the title compound in two steps: 49.4%.

3.4- (Piperidin-2-yl) butyrate hydrochloride preparation (Int-20)

To Int-19 (7.0 g,18.8 mmol) was added 6M aqueous hydrochloric acid (80 mL) and the mixture was allowed to react at 100℃for 20 hours. After the reaction is finished, the solvent is removed under reduced pressure, and the crude product of the target compound is obtained and is directly used for the next step.

4. Preparation of octahydro-4H-quinolin-4-one (Int-21)

To a solution of Int-20 (crude in the above step) in DCM (100 mL) was added TEA (11.4 g,112.6 mmol) and HATU (7.1 g,18.7 mmol) and reacted at 25℃for 1 h. After the reaction, the reaction was quenched with water, the organic phase was extracted with DCM, dried and concentrated and purified by silica gel column (EA: pe=7:3) to give the target compound 1.6g, two step yield: 55.4%.

5.3 Preparation of 3- (3-bromo-2- (methoxymethoxy) 5-methylbenzoyl) octahydro-4H-quinolin-4-one (Int-22)

NaHMDS (2M) (10.4 mL,20.8 mmol) was added dropwise to a solution of Int-8 (3.0 g,10.4 mmol) and Int-21 (1.6 g,10.4 mmol) in THF (50 mL) at-5℃for 5min, and then water (50 mL) was added thereto at 25℃for 10 min for 16 h. After the reaction, the organic phase was extracted with EA and the residue was concentrated by column chromatography (EA: pe=1:6) to give 3.6g of the target product, yield: 84.6%.

6.3 Preparation of 3- (3-bromo-2-hydroxy-5-methylbenzoyl) octahydro-4H-quinolin-4-one (Int-23)

To a solution of Int-22 (3.6 g,8.8 mmol) in DCM (18 mL) was added TFA (6 mL) and the reaction was allowed to proceed at 25℃for 2 hours. After the reaction, concentrating, adding saturated sodium bicarbonate to adjust the pH to be alkaline, extracting an organic phase with DCM, drying and concentrating to obtain a crude product of the target compound, which is directly used in the next step.

Preparation of 11-bromo-9-methyl-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen-o [3,2-c ] quinolin-7-one (Int-24)

To a solution of Int-23 (crude in the above step) in DCM (50 mL) was added trifluoromethanesulfonic anhydride (10 mL) and the reaction was continued at 25℃for 16 hours. After the reaction, the mixture was concentrated, saturated sodium bicarbonate solution was added to adjust ph=8, DCM was added to extract the organic phase, and the residue was concentrated by column chromatography (EA: pe=4:1) to give 2.0g of the target product, two step yield: 65.5%.

8.11 Preparation of acetyl-9-methyl-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen-o [3,2-c ] quinolin-7-one (Int-25)

To a solution of Int-24 (2.3 g,6.6 mmol) in dioxane (40 mL), int-37 (3.1 g,8.6 mmol) was added, pd (PPh ₃)₂Cl₂ (463 mg,0.66 mmol) was added, and reacted under nitrogen protection at 95℃for 16 hours, cooled to 50℃HCl (1M, 20 mL) was added, reacted for 0.5 hours, quenched with saturated potassium fluoride solution at 25℃and filtered, the organic phase was extracted by EA and the residue was concentrated by column chromatography (EA: PE=2:3) to give the target product 2.0g in 97.2% yield.

9.Preparation of 11- (1-hydroxyethyl) -9-methyl-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen-o [3,2-c ] quinolin-7-one (Int-26)

To a solution of Int-25 (350 mg,1.1 mmol) in methanol (5 mL) was added sodium borohydride (83 mg,2.2 mmol) and the mixture was reacted at 25℃for 1 hour. After the reaction, adding water for quenching, extracting with DCM, drying and concentrating to obtain the crude product of the target compound which is directly used in the next step.

Preparation of 11- (1-bromoethyl) -9-methyl-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen-o [3,2-c ] quinolin-7-one (Int-27)

To DCM (10 mL) of Int-26 (crude in the previous step) was added phosphorus tribromide (893 mg,3.3 mmol) at 0deg.C and reacted for 1 hour at 25deg.C. After the reaction, saturated sodium bicarbonate solution was added to adjust ph=8, DCM was added to extract the organic phase, and the residue was concentrated by column chromatography (EA: dcm=1:2) to give 250mg of the target product, two step yield: 59.1%.

11.2 Preparation of methyl- ((1- (9-methyl-7-oxo-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen [3,2-c ] quinolin-11-yl) ethyl) benzoate (Int-28)

To a solution of Int-27 (250 mg,0.66 mmol) in DMF (10 mL) was added Int-15 (150 mg,0.99 mmol) and DIPEA (256 mg,2.0 mmol) and reacted at 80℃for 16 hours. After the reaction, quench with water, extract the organic phase with EA, concentrate the residue on column chromatography (EA: dcm=1:2) to give 120mg of the desired product in yield: 40.5%.

12.2 Preparation of- ((1- (9-methyl-7-oxo-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen [3,2-c ] quinolin-11-yl) ethyl) amino) benzoic acid (Compound 2)

To a solution of Int-28 (200 mg,0.46 mmol) in a mixture of methanol (3 mL), water (3 mL) and THF (3 mL) was added lithium hydroxide monohydrate (168 mg,4.0 mmol) and reacted at 50℃for 4 hours. After the reaction, EA was added to extract the organic phase twice, ph=6 of the aqueous phase was adjusted with a dilute hydrochloric acid solution, the organic phase was extracted with DCM, and the product was dried and concentrated to give the objective product 30mg, yield: 15.1%.

Molecular formula C ₂₆H₂₈N₂O₄ molecular weight 432.5LC-MS (M/e): 433.2 (M+H ⁺)

¹H-NMR(400MHz,DMSO-d₆)δ:12.9-12.7(m,1H),8.35-8.33(m,1H),7.81-7.79(m,1H),7.61(s,1H),7.34(s,1H),7.24-7.23(m,1H),6.53-6.52(m,1H),6.47-6.43(m,1H),5.08-5.06(m,1H),4.03-4.01(m,1H),3.02-2.90(m,1H),2.70-2.60(m,1H),2.59-2.57(m,2H),2.29(s,3H),2.00-1.90(m,1H),1.85-1.65(m,4H),1.63-1.60(m,3H),0.25-0.15(m,3H).

Preparation example 3: preparation of 2- ((1- (2-methyl-12-oxo-7, 8,9,10 a, 11-hexahydro-12H-chromen-3, 2-b ] indolizin-4-yl) ethyl) amino) benzoic acid (Compound 3)

1. (E) Preparation of tert-butyl-2- (3-methoxy-3-oxopropyl-1-en-1-yl) piperidine-1-carboxylate (Int-30)

Tert-butyl 2-formylpiperidine-1-carboxylate (Int-29) (8.0 g,37.5 mmol) was dissolved in THF (80 mL), CMMTPP (12.5 g,37.5 mmol) was added and reacted at 70℃for 4 hours. The system was quenched with water, the organic phase was extracted with EA, and the concentrated residue was purified by a silica gel column (EA/PE=15%), to give the objective compound (9.5 g, yield: 94.1%).

2.Preparation of tert-butyl 2- (3-methoxy-3-oxopropyl) piperidine-1-carboxylate (Int-31)

Int-30 (9.5 g,35.2 mmol) was dissolved in methanol (100 mL), and wet palladium on carbon (950 mg) was added thereto and reacted at 25℃for 16 hours in a hydrogen atmosphere. Filtration, drying and concentration gave the objective compound (9.3 g, yield: 96.9%).

3.3 Preparation of methyl 3- (piperidin-2-yl) propionate (Int-32)

To Int-31 (7.0 g,25.7 mmol) in DCM (70 mL) at 25℃was added TFA (1 mL) and the reaction was allowed to proceed for 16 h. After the completion of the reaction, the solvent was removed under reduced pressure, the residue was diluted with DCM, saturated sodium bicarbonate was added to adjust the pH to alkaline, the organic phase was extracted with DCM methanol, dried, and concentrated to give the title compound (3.2 g, yield: 72.7%).

4. Preparation of hexahydroindolizine-3 (2H) -one (Int-33)

Int-32 (3.2 g,18.6 mmol) was dissolved in DMF (35 mL), and potassium carbonate (12.8 g,93.0 mmol) was added and reacted at 80℃for 16 hours. After the reaction, water was added to quench the reaction, EA was added to extract the reaction mixture, and the mixture was concentrated to obtain a crude product (2.5 g).

5.Preparation of 2- (3-bromo-2- (methoxymethoxy) -5-methylbenzoyl) indolizin-3 (2H) -one (Int-34)

Int-33 (2.5 g,17.9 mmol) was dissolved in THF (50 mL), int-8 (5.2 g,17.9 mmol) was added thereto, lithium bis (trimethylsilyl) amide (36 mL,36 mmol) was added dropwise at 25℃and reacted at 25℃for 30 minutes, water (40 mL) was added and reacted at 25℃for 16 hours. After the completion of the reaction, the mixture was extracted twice with EA, washed once with water, and the residue was concentrated by column chromatography (EA/n-heptane=45%) to give the objective product (4.5 g, yield: 63.4%)

6.Preparation of 2- (3-bromo-2-hydroxy-5-methylbenzoyl) hexahydroindolizin-3 (2H) -one (Int-35)

Int-34 (4.5 g,11.4 mmol) was dissolved in DCM (50 mL) and TFA (10 mL) was added and reacted at 25℃for 3 hours. After the reaction, the mixture was concentrated, saturated sodium bicarbonate was added to adjust ph=8, ea was extracted twice, and dried, and concentrated to give the objective compound (4.2 g crude product).

Preparation of 4-bromo-2-methyl-7, 8,9,10 a, 11-hexahydro-12H-chromen [3,2-b ] indolizin-12-one (Int-36)

A solution of Int-35 (4.2 g of crude) in DCM (50 mL) was added to the solution at 25℃with trifluoromethanesulfonic anhydride (10 mL) and the reaction was continued at 25℃for 16 h. After the reaction, the mixture was concentrated, and saturated sodium bicarbonate solution was added to adjust ph=8, DCM was added to extract, and the mixture was washed once with water and concentrated to give the objective compound (3.6 g, two-step yield: 94.7%).

8.4 Preparation of acetyl-2-methyl-7, 8,9,10 a, 11-hexahydro-12H-chromen-o [3,2-b ] indolizin-12-one (Int-38)

Int-36 (3.6 g,10.8 mmol) was dissolved in dioxane (40 mL), int-37 (4.7 g,13.0 mmol) was added, pd (PPh ₃)₂Cl₂ (1.5 g,2.16 mmol) was added, reacted under nitrogen protection at 95℃for 16 hours, HCl (1M, 40 mL) was added at 50℃for 0.5 hours, saturated potassium fluoride solution was added at 25℃for 0.5 hours, filtration, EA extraction once with DCM, concentrated residue column chromatography (EA/n-heptane=40%) gave the target product (2.0 g, yield: 62.5%)

9.Preparation of 4- (1-hydroxyethyl) -2-methyl-7, 8,9,10 a, 11-hexahydro-12H-chromen-o [3,2-b ] indolizin-12-one (Int-39)

Int-38 (700 mg,2.3 mmol) was dissolved in methanol (10 mL), and sodium borohydride (133 mg,3.5 mmol) was added thereto and reacted at 25℃for 1 hour. After the reaction, water was added to quench, DCM was extracted, dried and concentrated to give the title compound (600 mg, 85.1%).

Preparation of 4- (1-bromoethyl) -2-methyl-7, 8,9,10 a, 11-hexahydro-12H-chromen-o [3,2-b ] indolizin-12-one (Int-40)

Int-39 (600 mg,2.0 mmol) was dissolved in DCM (10 mL) and phosphorus tribromide (1.6 g,6.0 mmol) was added at 0deg.C and reacted at 30deg.C for 4 hours. After the reaction was completed, a saturated sodium bicarbonate solution was added to adjust ph=8, DCM was added to extract twice, and the residue was concentrated by column chromatography (methanol/dcm=15%) to give the objective product (660 mg, yield: 91.4%).

11.2 Preparation of methyl- ((1- (2-methyl-12-oxo-7, 8,9,10 a, 11-hexahydro-12H-chromen-3, 2-b ] indolizin-4-yl) ethyl) amino) benzoate (Int-41)

Int-40 (450 mg,1.2 mmol) was dissolved in DMF (10 mL), int-15 (283 mg,1.9 mmol) was added, DIPEA (460 mg,3.6 mmol) was added and the reaction was carried out at 80℃for 3 hours. After the reaction, it was quenched with water, extracted twice with EA, and the residue was concentrated by column chromatography (methanol/dcm=16%) to give the target product (381 mg, yield: 70.8%).

12.2 Preparation of- ((1- (2-methyl-12-oxo-7, 8,9,10 a, 11-hexahydro-12H-chromen-o [3,2-b ] indolizin-4-yl) ethyl) amino) benzoic acid (Compound 3)

Int-41 (200 mg,0.46 mmol) was dissolved in methanol (5 mL) and water (5 mL), lithium hydroxide monohydrate (10 mL) was added at 25℃and reacted at 40℃for 3 hours. After the reaction, pH=5 was adjusted by adding 2M hydrochloric acid solution, and then extracted with EA, and the residue was concentrated by column chromatography (methanol/water=55%), followed by beating with EA to obtain the objective product (12 mg, yield: 6.2%).

Molecular formula C ₂₅H₂₆N₂O₄ molecular weight 418.5LC-MS (M/e): 419.2 (M+H ⁺)

¹H-NMR(400MHz,DMSO-d₆)δ:12.68(s,1H),8.53(s,1H),7.80-7.78(m,1H),7.63(s,1H),7.34(s,1H),7.22-7.18(m,1H),6.53-6.46(m,2H),5.08-5.05(m,1H),4.02(m,1H),3.84-3.82(m,1H),3.08-3.06(m,2H),2.50(m,1H),2.40-2.31(m,3H),1.81-1.70(m,3H),0.56-0.62(m,3H),0.51-0.46(m,3H).

Preparation example 4: preparation of 2- ((1- (2-fluoro-8-methyl-10-oxo-10, 11-indano [1,2-b ] chromen-6-yl) ethyl) amino) benzoic acid (Compound 4)

1.3 Preparation of (Int-44) propionic acid (2-bromo-4-methylphenoxy)

To a solution of 2-bromo-4-methylphenol (Int-42) (25.0 g,134.4 mmol) in DMF (500 mL) at 25℃was added 3-bromopropionic acid (Int-43) (20.4 g,134.4 mmol), and the reaction was carried out at 25℃for 2 hours. After the reaction, 2M hydrochloric acid solution (200 mL) was added for quenching, EA (300 mL x 3) was added for extraction, the organic phase was collected, dried by spin, and column chromatography (SiO ₂, PE: ea=1:1) to give 23.0g of product, yield: 66.3%.

Preparation of 8-bromo-6-methylchromen-4-one (Int-45)

To Int-44 (23.0 g,0.089 mol) was added p-toluenesulfonic acid (100 mL) at 25℃and reacted at 40℃for 1 hour. After the reaction, the reaction mixture was slowly added dropwise to ice water (200 mL) to quench the reaction, EA (200 ml×3) was extracted, the organic phase was collected, dried by spin, and column chromatography (SiO ₂, PE: EA3: 1) gave 18.0g of product, yield: 84.3%.

3. (E) Preparation of 8-bromo-3- (2-bromo-5-fluorobenzylidene) -6-methylchromen-4-one (Int-47)

To Int-45 (18.0 g,75.0 mmol) ethanol (200 mL) was added 2-bromo-5-fluorobenzaldehyde (Int-46) (22.7 g,112.5 mmol), 5% aqueous sodium hydroxide (50 mL) and stirred for 2h at 25℃and after completion of the reaction, the reaction mixture was filtered and the filter cake was washed with cold ethanol (20 mL. Times.3). After drying, 18.0g of product is obtained, the yield is 56.6%

4.8 Preparation of bromo-3- (2-bromo-5-fluorobenzyl) -6-methyl-4H-chromen-4-one (Int-48)

Int-47 (9.0 g,21.2 mmol) was dissolved in DMF (100 mL), potassium carbonate (4.4 mg,31.8 mmol) was added, and after the reaction was completed at 110 ℃ for 2 hours, the reaction mixture was dried by spin-drying, and the residue was purified by silica gel column (PE: ea=50:1) to give 3.0g of the target compound, yield: 33.3%.

Preparation of 8-acetyl-3- (2-bromo-5-fluorobenzyl) -6-methyl-4H-chromen-4-one (Int-49)

Int-48 (3.0 g,7.1 mmol) was dissolved in 1, 4-dioxane (50 mL), int-37 (2.6 g,7.1 mmol) was added, pd (PPh ₃)₂Cl₂ (498 mg,0.71 mmol), reacted at 95℃for 12 hours under nitrogen atmosphere, 2M HCl aq. (40 mL) was added after the completion of the reaction, reacted at 50℃for 1 hour, extracted twice with EA, washed once with water, concentrated, and the residue was subjected to column chromatography (EA/n-heptane=1/5) to give 2.0g of the title compound in a yield of 72.6%.

6.3 Preparation of- (2-bromo-5-fluorobenzyl) -8- (1-hydroxyethyl) -6-methyl-4H-chromen-4-one (Int-50)

Int-49 (800 mg,2.1 mmol) was dissolved in methanol (20 mL), and sodium borohydride (238 mg,6.3 mmol) was added. The reaction was carried out at 25℃for 1 hour. After the reaction is finished, adding water to quench the reaction, adding EA to extract, collecting an organic phase, and concentrating to obtain 1.0g of crude product which is directly used for the next reaction.

7.3 Preparation of- (2-bromo-5-fluorobenzyl) -8- (1-bromoethyl) -6-methyl-4H-chromen-4-one (Int-51)

Int-50 (1.0 g, crude) was dissolved in DCM (20 mL) and phosphorus tribromide (1.7 g,6.3 mmol) was added. The reaction was quenched by addition of saturated aqueous sodium bicarbonate (50 mL) at 25 ℃ for 3h, extracted with DCM (50 mL x 3), and the organic phase collected, dried by spinning and the residue was chromatographed (SiO ₂, PE: ea=10:1) to give 470mg of the title compound in two steps with a yield of 49.5%.

8.2 Preparation of tert-butyl- ((1- (3- (2-bromo-5-fluorobenzyl) -6-methyl-4-oxo-4H-chromen-8-yl) ethyl) amino) benzoate (Int-52)

Int-51 (460 mg,1.0 mmol) was dissolved in acetonitrile (20 mL), and tert-butyl 2-aminobenzoate (212 mg,1.1 mmol) and N, N-diisopropylethylamine (387 mg,3.0 mmol) were added and reacted at 55℃for 12 hours. Spin-drying the reaction mixture and column chromatography of the residue (SiO ₂, PE: ea=50:1) gave 400mg of the title compound in a yield of 70.8%.

9.2 Preparation of tert-butyl- ((1- (2-fluoro-8-methyl-10-oxo-10, 11-indano [1,2-b ] chromen-6-yl) ethyl) amino) benzoate (Int-53)

Int-52 (200 mg,0.36 mmol) was dissolved in toluene (20 mL), palladium acetate (18 mg,0.072 mmol), johnPhos (42 mg,0.14 mmol) and potassium acetate (106 mg,1.1 mmol) were added, the reaction was carried out at 110℃for 2 hours under nitrogen atmosphere, the reaction mixture was dried by spin-drying, and the residue was subjected to silica gel column chromatography (EA/n-heptane=1/20) to give the objective compound, 40mg, yield: 22.9%.

10.2 Preparation of- ((1- (2-fluoro-8-methyl-10-oxo-10, 11-indano [1,2-b ] chromen-6-yl) ethyl) amino) benzoic acid (Compound 4)

Int-53 (40 mg,0.082 mmol) was dissolved in DCM (2 mL), TFA (2 mL) was added and the reaction was allowed to react at 40℃for 3 hours, the reaction was dried by spin-drying, and the residue was chromatographed on a reverse phase column (water/acetonitrile=5/3) to give the title compound 15mg, yield: 42.5%.

Molecular formula C ₂₆H₂₀FNO₄ molecular weight 429.1LC-MS (M/e): 430.1 (M+H ⁺)

¹H-NMR(400MHz,DMSO-d₆)δ:12.85(brs,1H),8.51(d,J＝6.0Hz,1H),8.04-8.01(m,1H),7.82(s,1H),7.79(d,J＝7.7Hz,1H),7.58(s,2H),7.39(t,J＝8.8Hz,1H),7.20(t,J＝6.5Hz,1H),7.00(d,J＝51.3Hz,1H),6.72-6.50(m,1H),6.50-6.38(m,1H),5.41-5.35(m,1H),,2.37(s,3H),1.66(d,J＝6.1Hz,3H).

Preparation example 5: preparation of 2- ((1- (8-methyl-6-oxo-1, 2, 3a,4, 5-hexahydro-6H-chromen-o [2,3-e ] indolizin-10-yl) ethyl) amino) benzoic acid (Compound 10)

Preparation of tert-butyl 2- (2-hydroxyethyl) pyrrolidine-1-carboxylate (Int-55)

To a solution of 2- (1- (tert-butoxycarbonyl) pyrrolidin-2-yl) acetic acid (Int-54) (10.0 g,43.6 mmol) in THF (100 mL) was added a solution of borane in tetrahydrofuran (8.8 g,87.0 mmol) at 25℃followed by reaction at 40℃for 6 hours. The system was quenched with methanol, the mixture was concentrated in the organic phase, the crude product was extracted with water, and the organic phase was concentrated to dryness in vacuo to give 9.5g of the title compound in 101.0% yield.

2.Preparation of tert-butyl 2- (2- ((methylsulfonyl) oxy) ethyl) pyrrolidine-1-carboxylate (Int-56)

To a solution of Int-55 (5.4 g,25.1 mmol) in DCM (60 mL) was added TEA (5.1 g,50.4 mmol) and methylsulfonyl chloride (4.3 g,37.5 mmol) dropwise at 0deg.C for 30 min. The system was quenched with water, the organic phase extracted with DCM, concentrated and spun-dried to give crude product which was used directly in the next step.

3.Preparation of diethyl 2- (2- (1- (tert-Butoxycarbonyl) pyrrolidin-2-yl) ethyl) malonate (Int-58)

To a solution of diethyl malonate (Int-57) (7.5 g,49.9 mmol) in THF (100 mL) at 0 ℃ was added sodium hydride (60%) (2.0 g,50.0 mmol), and the mixture was reacted for 30 minutes, then Int-56 (crude product of the above step) was added, and the mixture was reacted at 55 ℃ for 24 hours. The pH was adjusted to 4 with 1M dilute hydrochloric acid at 0deg.C, the organic phase was extracted with EA, dried and concentrated, and purified by silica gel column (EA: PE=1:4) to give 3.2g of the target compound, two step yield: 35.7%.

4.Preparation of 4- (pyrrolidin-2-yl) butyrate hydrochloride (Int-59)

To Int-58 (3.2 g,9.0 mmol) was added 6M aqueous hydrochloric acid (50 mL) and the mixture was allowed to react at 100℃for 16 hours. After the reaction is finished, the solvent is removed under reduced pressure, and the crude product of the target compound is obtained and directly used for the next step.

5. Preparation of hexahydroindolizine-5 (1H) -one (Int-60)

To a solution of Int-59 (crude in the above step) in DCM (50 mL) was added TEA (2.7 g,26.7 mmol) and HATU (3.4 g,8.9 mmol) and reacted at 25℃for 1 h. After the reaction, the reaction was quenched with water, the organic phase was extracted with DCM, dried and concentrated and purified by silica gel column (EA: pe=7:3) to give 1.5g of crude target compound.

6.6 Preparation of 6- (3-bromo-2- (methoxymethoxy) -5-methylbenzoyl) indolizin-5 (1H) -one (Int-61)

To a solution of Int-8 (3.0 g,10.4 mmol) and Int-60 (1.5 g crude) in THF (50 mL) at-5 ℃ was added NaHMDS (2M) (10.4 mL,20.8 mmol) dropwise, reacted for 5 min, reacted at 25 ℃ for 10min, then water and EA were added to extract the organic phase, and the residue was concentrated by column chromatography (EA: pe=1:2) to give the target product 1.3g, yield: 31.6%.

7.6 Preparation of 6- (3-bromo-2-hydroxy-5-methylbenzoyl) hexahydroindolizin-5 (1H) -one (Int-62)

To a solution of Int-61 (1.3 g,3.3 mmol) in DCM (25 mL) was added TFA (5 mL) and the reaction was allowed to proceed at 25℃for 1 hour. After the reaction, concentrating, adding saturated sodium bicarbonate to adjust the pH to be alkaline, extracting an organic phase with DCM, drying and concentrating to obtain a crude product of the target compound, which is directly used in the next step.

Preparation of 10-bromo-8-methyl-1, 2, 3a,4, 5-hexahydro-6H-chromen [2,3-e ] indolizin-6-one (Int-63)

To a solution of Int-62 (crude in the above step) in DCM (60 mL) was added trifluoromethanesulfonic anhydride (15 mL) and the reaction was continued at 25℃for 10 hours. After the reaction, the mixture was concentrated, ph=8 was adjusted by adding saturated sodium bicarbonate solution, the organic phase was extracted with DCM, and the residue was concentrated by column chromatography (EA: pe=7:3) to give 550mg of the target product in two steps: 35.3%.

9.10 Preparation of acetyl-8-methyl-1, 2, 3a,4, 5-hexahydro-6H-chromen-o [2,3-e ] indolizin-6-one (Int-64)

To a solution of Int-63 (550 mg,1.6 mmol) in dioxane (25 mL), int-37 (751 mg,2.1 mmol) was added, pd (PPh ₃)₂Cl₂ (112 mg,0.16 mmol) was added, and reacted at 95℃for 16 hours under nitrogen protection, HCl (1M, 5 mL) was added to cool to 50℃and reacted for 0.5 hours, quenched with saturated potassium fluoride solution at 25℃and filtered, the organic phase was extracted by EA, and the residue was concentrated by column chromatography (EA: DCM=4:1) to give 430mg of the desired product in 87.9% yield.

10.Preparation of 10- (1-hydroxyethyl) -8-methyl-1, 2, 3a,4, 5-hexahydro-6H-chromen-o [2,3-e ] indolizin-6-one (Int-65)

To a solution of Int-64 (400 mg,1.3 mmol) in methanol (15 mL) was added sodium borohydride (147 mg,3.9 mmol) and the mixture was reacted at 25℃for 1 hour. After the reaction, water is added to catalyze, DCM is used for extraction, drying and concentration are carried out to obtain 400mg of crude target compound.

11.10 Preparation of (1-bromoethyl) -8-methyl-1, 2, 3a,4, 5-hexahydro-6H-chromen-o [2,3-e ] indolizin-6-one (Int-66)

To DCM (20 mL) of Int-65 (200 mg, crude in the previous step) was added phosphorus tribromide (544 mg,2.0 mmol) at 0deg.C and reacted for 2 hours at 25deg.C. After the reaction, saturated sodium bicarbonate solution was added to adjust ph=8, DCM was added to extract the organic phase, and the residue was concentrated by column chromatography (EA: dcm=4:1) to give 58mg of the target product, two step yield: 23.8%.

12.2 Preparation of methyl- ((1- (8-methyl-6-oxo-1, 2, 3a,4, 5-hexahydro-6H-chromen-2, 3-e ] indolizin-10-yl) ethyl) benzoate (Int-67)

To a solution of Int-66 (58 mg,0.16 mmol) in DMF (5 mL) was added Int-15 (150 mg,0.24 mmol) and DIPEA (62 mg,0.48 mmol) and reacted at 80℃for 16 hours. After the reaction, quench with water, extract the organic phase with EA, concentrate the residue on column chromatography (EA: dcm=7:3) to give 40mg of the desired product in yield: 57.8%.

13.2 Preparation of- ((1- (8-methyl-6-oxo-1, 2, 3a,4, 5-hexahydro-6H-chromen-o [2,3-e ] indolizin-10-yl) ethyl) amino) benzoic acid (Compound 10)

To a solution of Int-67 (40 mg,0.092 mmol) in methanol (3 mL), water (3 mL) and THF (3 mL) was added lithium hydroxide monohydrate (77 mg,1.8 mmol) and reacted at 50℃for 4 hours. After the reaction, EA was added to extract the organic phase twice, ph=6 of the aqueous phase was adjusted with a dilute hydrochloric acid solution, the organic phase was extracted with DCM, and the resultant was dried and concentrated to give the objective product 17mg, yield: 43.9%.

¹H-NMR(400MHz,DMSO-d6)δ:12.8-12.7(m,1H),8.37-8.33(m,1H),7.80-7.78(m,1H),7.61(s,1H),7.31-7.21(m,2H),6.55-6.44(m,2H),5.08-5.03(m,1H),3.80-3.73(m,1H),3.60-3.58(m,2H),2.86-2.82(m,1H),2.29(s,3H),2.25-1.85(m,5H),1.75-1.65(m,3H),1.35-1.15(m,2H).

Preparation example 6: preparation of 2- ((1- (3-fluoro-9-methyl-7-oxo-5, 7-dihydro-6H-benzo [ c ] xanthen-11-yl) ethyl) amino) benzoic acid (Compound 12)

1.8 Preparation of bromo-6-methyl-4-oxo-4H-chromene-3-carbaldehyde (Int-71)

1- (3-Bromo-2-hydroxy-5-methylphenyl) ethan-1-one (Int-70) (4.5 g,19.3 mmol) was dissolved in N, N-dimethylformamide (50 mL), phosphorus oxychloride (5 mL) was added, and the mixture was reacted at 25℃for 16 hours. After the reaction is finished, adding water into the system to separate out a large amount of solids, filtering, and spin-drying to obtain 4.0g of target compound with the yield: 77.7%.

2. Preparation of (2-bromo-5-fluorobenzylidene) triphenyl-lambda ⁵ -phosphine (Int-69)

1-Bromo-2- (bromomethyl) -4-fluorobenzene (Int-68) (4.0 g,14.9 mmol) was dissolved in toluene (50 mL), triphenylphosphine (4.7 g,17.9 mmol) was added, reacted at 120℃for 3 hours, potassium t-butoxide (3.0 g,26.8 mmol) was added at 25℃and reacted at 25℃for 30 minutes to give 55mL toluene reaction solution which was directly used in the next step.

3. (E) Preparation of 8-bromo-3- (2-bromo-5-fluorostyryl) -6-methyl-4H-chromen-4-one (Int-72)

To 55mL of the toluene reaction solution obtained in the previous step was added Int-71 (4.0 g,14.9 mmol), and toluene (20 mL) was added thereto and the mixture was reacted at 100℃for 1 hour. After the reaction, quench with water, extract with EA twice, concentrate, and purify the residue on a silica gel column (EA/pe=20%), to give 3.2g of the target compound, yield: 49.2%.

4.8 Preparation of bromo-3- (2-bromo-5-fluorophenylethyl) -6-methyl-4H-chromen-4-one (Int-73)

Int-72 (3.2 g,7.3 mmol) was dissolved in methanol (15 mL) and THF (15 mL), and triphenylphosphine rhodium chloride (340 mg,0.73 mmol) was added and reacted under hydrogen atmosphere at 25℃for 18 hours. After the reaction, the residue was directly concentrated by column chromatography (EA/pe=20%) to give the objective product 2.2g, yield: 68.8%.

Preparation of 11-bromo-3-fluoro-9-methyl-5, 6-dihydro-7H-benzo [ c ] xanthen-7-one (Int-74)

Int-73 (2.2 g,5.0 mmol) was dissolved in DMF (25 mL), and palladium acetate (225 mg,1.0 mmol), potassium acetate (982 mg,10.0 mmol) and (2-biphenylyl) di-tert-butylphosphine (299 mg,1.0 mmol) were added and reacted at 95℃for 2 hours under nitrogen. After the completion of the reaction, the reaction was quenched with water, extracted twice with EA, concentrated, and the residue was purified by silica gel column (EA/pe=25%) to give 300mg of the objective compound in 16.7% yield.

Preparation of 11-acetyl-3-fluoro-9-methyl-5, 6-dihydro-7H-benzo [ c ] xanthen-7-one (Int-75)

Int-74 (300 mg,0.84 mmol) was dissolved in dioxane (5 mL), int-37 (361 mg,1.0 mmol) and Pd (PPh ₃)₂Cl₂ (56 mg,0.08 mmol) were added and reacted at 95℃for 16 hours under nitrogen protection, 2M HCl solution (6 mL) was added at 50℃and reacted at 50℃for 50 minutes after completion of the reaction, quenched with saturated sodium bicarbonate solution, extracted twice with EA, concentrated and purified by silica gel column (EA/PE=30%) to give 140mg of the title compound in a yield of 51.9%.

Preparation of 3-fluoro-11- (1-hydroxyethyl) -9-methyl-5, 6-dihydro-7H-benzo [ c ] xanthen-7-one (Int-76)

Int-75 (140 mg,0.43 mmol) was dissolved in methanol (5 mL), and sodium borohydride (25 mg,0.65 mmol) was added to react at 25℃for 1 hour. After the reaction, water was added to quench the reaction, DCM was used for extraction, and the reaction mixture was dried and concentrated to give 120mg of the target compound in 85.1% yield.

8.Preparation of 11- (1-bromoethyl) -3-fluoro-9-methyl-5, 6-dihydro-7H-benzo [ c ] xanthen-7-one (Int-77)

Int-76 (120 mg,0.37 mmol) was dissolved in DCM (5 mL) and phosphorus tribromide (300 mg,1.11 mmol) was added at 0deg.C and reacted at 30deg.C for 4 hours. After the reaction, saturated sodium bicarbonate solution is added to adjust the pH to be 8, and the mixture is concentrated to obtain 1000mg of target product with the yield: 69.9%.

9.2 Preparation of methyl- ((1- (3-fluoro-9-methyl-7-oxo-5, 7-dihydro-6H-benzo [ c ] xanthen-11-yl) ethyl) amino) benzoate (Int-78)

Int-77 (100 mg,0.26 mmol) was dissolved in DMF (5 mL), int-15 (59 mg,0.39 mmol) was added, DIPEA (101 mg,0.78 mmol) was added, and the reaction was carried out at 80℃for 3 hours. After the reaction, quench with water, extract with EA twice, concentrate the residue column chromatography (EA/pe=15%) to give the target product 40mg, yield: 34.0%.

10.2 Preparation of- ((1- (3-fluoro-9-methyl-7-oxo-5, 7-dihydro-6H-benzo [ c ] xanthen-11-yl) ethyl) amino) benzoic acid (Compound 12)

Int-78 (40 mg,0.087 mmol) was dissolved in methanol (2 mL) and water (2 mL), lithium hydroxide monohydrate (73 mg,1.75 mmol) was added at 25℃and reacted at 40℃for 3 hours. After the reaction, 2M hydrochloric acid solution was added to adjust ph=5, EA was added to extract, and the residue was concentrated by column chromatography (methanol/water=35%) to give 4mg of the objective product, yield: 11.0%.

Molecular formula C ₂₇H₂₂FNO₄ molecular weight 443.5LC-MS (M/e): 444.2 (M+H ⁺)

¹H-NMR(400MHz,DMSO-d₆)δ:8.06-8.04(m,1H),8.53(s,1H),7.92-7.91(m,1H),7.86(s,1H),7.59-7.58(m,1H),7.19-7.13(m,3H),6.57-6.48(m,2H),5.38-5.33(m,1H),3.04-3.00(m,2H),2.92-2.88(m,2H),2.39(s,3H),1.73-1.59(m,3H).

Preparation example 7: preparation of 3- (((1R) -1- (9-methyl-7-oxo-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen-o [3,2-c ] quinolin-11-yl) ethyl) amino) -6- (trifluoromethyl) picolinic acid (Compound 15)

Preparation of methyl 3-fluoro-6- (trifluoromethyl) picolinate (Int-80)

To a solution of 2-bromo-3-fluoro-6- (trifluoromethyl) pyridine (Int-79) (600 mg,2.5 mmol) in methanol (15 mL) and DMF (5 mL) were added Pd (dppf) Cl ₂ (181 mg,0.25 mmol) and TEA (506 mg,5.0 mmol), and the mixture was reacted at 60 ℃ for 16 hours under carbon monoxide atmosphere, water was added to the system, the organic phase was extracted by EA, and the residue was concentrated by column chromatography (EA: pe=1:10) to give 280mg of the objective product, yield: 51.0%.

2. Preparation of (R) -2-methyl-N- ((E) -1- (9-methyl-7-oxo-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen [3,2-c ] quinolin-11-yl) ethylene) propane-2-thioamide (Int-81)

To a solution of Int-25 (1.0 g,3.2 mmol) in THF (50 mL) was added (R) -2-methylpropane-2-sulfonamide (776 mg,6.4 mmol) and tetraisopropyl titanate (3.6 g,12.7 mmol) and reacted at 75℃for 32 hours. After the reaction, the reaction mixture was quenched with water, filtered, the organic phase extracted with EA, dried and the residue concentrated by column chromatography (EA: dcm=1:2) to give 1.0g of the desired product in yield: 75.7%.

3. Preparation of (R) -2-methyl-N- ((1R) -1- (9-methyl-7-oxo-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen [3,2-c ] quinazolin-11-yl) ethyl) propane-2-thioamide (Int-82)

To a solution of Int-81 (750 mg,1.4 mmol) in methanol (15 mL) at-78deg.C were added cerium chloride heptahydrate (261 mg,0.70 mmol) and sodium borohydride (106 mg,2.8 mmol), and the mixture was reacted at-78deg.C for 4 hours. After the reaction, water was added to the system, the organic phase was extracted with DCM, and the concentrated residue was purified in reverse phase (methanol: water=3:2) to give 470mg of the desired product, yield: 82.1%.

4.11- ((R) -1-aminoethyl) -9-methyl-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen-o [3,2-c ] quinolin-7-one preparation (Int-83)

To a solution of Int-82 (200 mg,0.48 mmol) in EA (10 mL) was added a solution of EA hydrochloride (1.0 mL) and the mixture was reacted at 25℃for 1 hour. The reaction solution was dried by spinning, diluted with DCM, pH was adjusted to alkaline with saturated sodium bicarbonate solution, the organic phase was extracted with DCM, and concentrated to dryness to give crude product of the desired product, 190mg.

Preparation of methyl 3- (((1R) -1- (9-methyl-7-oxo-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen [3,2-c ] quinolin-11-yl) ethyl) amino) -6- (trifluoromethyl) picolinate (Int-84)

To a solution of Int-83 (135 mg,0.43 mmol) in DMF (10 mL) was added Int-80 (144 mg,0.66 mmol) and DIPEA (278 mg,2.2 mmol) and reacted at 100℃for 16 hours. After the reaction, quench with water, extract the organic phase with EA, concentrate the residue on column chromatography (EA: dcm=1:3) to give 115mg of the target product, yield: 51.6%.

Preparation of 3- (((1R) -1- (9-methyl-7-oxo-2, 3, 4a,5, 6-hexahydro-1H, 7H-chromen [3,2-c ] quinolin-11-yl) ethyl) amino) -6- (trifluoromethyl) picolinic acid (Compound 15)

To a solution of Int-84 (95 mg,0.18 mmol) in methanol (4 mL), water (4 mL) and THF (4 mL) was added lithium hydroxide monohydrate (151 mg,3.6 mmol) and reacted at 50℃for 30 minutes. After the reaction, EA was added to extract the organic phase twice, ph=6 of the aqueous phase was adjusted with a dilute hydrochloric acid solution, the organic phase was extracted with DCM, and the product was dried and concentrated to obtain 20mg of the objective product, yield: 21.6%.

Molecular formula C ₂₆H₂₆F₃N₃O₄ molecular weight 501.5LC-MS (M/e): 502.2 (M+H ⁺)

¹H-NMR(400MHz,DMSO-d6)δ:13.7-13.0(m,1H),8.65-8.55(m,1H),7.72-7.69(m,1H),7.64(s,1H),7.38(s,1H),7.12-7.08(m,1H),5.15-5.05(m,1H),4.40-4.28(m,1H),3.40-3.30(m,2H),3.02-2.88(m,1H),2.65-2.55(m,1H),2.50-2.45(m,1H),2.30(s,3H),1.98-1.92(m,1H),1.76-1.73(m,3H),1.67-1.63(m,4H),1.47-1.43(m,2H).

Using the same or similar methods as the preparation examples described above, the compounds shown in the following tables were prepared:

2 test of pharmacological Activity of Compounds of the invention

The beneficial effects of the compounds of the present invention are further illustrated below by means of pharmacological test examples, but this should not be understood as the compounds of the present invention having only the following beneficial effects.

Experimental example 1 in vitro cytological inhibitory Activity of the Compounds of the invention

Test objects part of the compounds according to the invention are given in the preparation examples for their chemical names and structures.

The cell lines used in the following experiments were as follows:

T47D: PI3K alpha H1047R mutant human breast ductal carcinoma cells

MDA-MB-453: PI3K alpha H1047R mutant human breast cancer cells

ZR-75-1: PI3K alpha WT wild type human breast cancer cells

Experimental method (CelltiterGlo assay)

1. Preparation of cells

1.1 Cell culture:

All cells were adherent cells, the T47D and ZR-75-1 cell culture media were RPMI Medium 1640+10% FBS+1% PS, the MDA-MB-453 cell culture media were DMEM Medium+10% FBS+1% PS, and the cells were tested in the logarithmic phase.

1.2 Preparation of cell suspension:

Cells in the logarithmic growth phase were harvested and counted using a platelet counter. Cell viability was checked by trypan blue exclusion, ensuring that cell viability was above 90%. To adjust to the appropriate concentration, add 90. Mu.L cell suspension to a white bottom transparent 96-well plate, and incubate for 24h.

TABLE 1 cell seed number

2. Formulation of test compounds

2.1 Preparation of test compound DMSO stock solution at a concentration of 10mM.

2.2 Preparing test Compound working solution

The test compound stock solution was 10mM, diluted 3-fold with DMSO, and 8 concentrations in total. Then 2 mu L of the gradient diluted compound is added into 198 mu L of culture solution to obtain test compound working solution (the concentration of the compound working solution is 10 times of the final concentration, and the highest concentration is 100 mu M)

2.3 Compound treatment

24H after inoculation of the cells, 10. Mu.L of compound working solution (10-fold dilution, final DMSO concentration of 0.1%) was added to each well, and the final concentration of test compound was: 10. Mu.M, 3.3. Mu.M, 1.1. Mu.M, 0.37. Mu.M, 0.123. Mu.M, 0.041. Mu.M, 0.014. Mu.M, 0.005. Mu.M, solvent control wells were added with 10. Mu.L of 1% DMSO solution and blank wells contained no cell-only medium.

2.4 Control well settings

Solvent control: 0.1% DMSO.

Blank control: and (5) a culture solution.

2.5 96 Well plates to which test compounds have been added are incubated for 3 days at 37℃in a 5% CO ₂ cell incubator.

3. Detection of

The CTG reagent was thawed at room temperature and the 96-well plate was equilibrated to room temperature for 30 minutes, 60. Mu.L of reagent (CELLTITER GLO ASSAY KIT) was added to each well, shaken for 2 minutes with a shaker (light-shielding), and incubated at room temperature for 20 minutes (light-shielding). The multifunctional enzyme label instrument reads the optical signal value.

4. Data processing

1) Inhibition (%) = [1- (test compound well reading-blank control well reading)/(DMSO solvent control well reading-blank control well reading) ]x100%;

2) IC ₅₀ values were plotted and calculated based on inhibition.

Experimental results and conclusions

TABLE 2 in vitro cytological Activity of the Compounds of the invention

As can be seen from Table 2, the compounds of the present invention can effectively inhibit proliferation of T47D, MDA-MB-453 cells with PI3K alpha H1047R mutation, while having no proliferation inhibition activity on ZR-75-1 cells of PI3K alpha WT, indicating that the compounds of the present invention can effectively and selectively inhibit proliferation of PI3K alpha H1047R mutant tumor cells.

Experimental example 2 in vitro cytological inhibitory Activity of the Compounds of the invention

The cell lines used in the following experiments were as follows:

T47D: PI3K alpha H1047R mutant human breast ductal carcinoma cells

MDA-MB-453: PI3K alpha H1047R mutant human breast cancer cells

Experimental method (CelltiterGlo assay)

1. Preparation of cells

1.1 Cell culture:

all cells were adherent cells, T47D cell culture Medium was RPMI Medium 1640+10% FBS+1% PS, MDA-MB-453 cell culture Medium was DMEM Medium+10% FBS+1% PS, and cells were tested in the logarithmic phase.

1.2 Preparation of cell suspension:

cells in the logarithmic growth phase were harvested and counted using a platelet counter. Cell viability was checked by trypan blue exclusion, ensuring that cell viability was above 90%. To adjust to the appropriate concentration, add 90. Mu.L cell suspension to a white bottom transparent 96-well plate, and incubate for 24h. One of the plates was plated with cells alone, and CTG was detected after 24 hours of culture as a cell viability base value control D0.

TABLE 3 cell seeding number

2. Formulation of test compounds

2.2 Preparing test Compound working solution

The test compound stock solution was 10mM, diluted 3-fold with DMSO, and 8 concentrations in total. Then, 2. Mu.L of the gradient diluted compound was added to 198. Mu.L of the culture solution, respectively, to give test compound working solutions (the concentration of the compound working solution was 10 times the final concentration, and the maximum concentration was 100. Mu.M).

2.3 Compound treatment

2.4 Control well settings

Solvent control: 0.1% DMSO.

Blank control: and (5) a culture solution.

2.5 96 Well plates to which test compounds have been added are incubated for 5 days at 37℃in a 5% CO ₂ cell incubator.

3. Detection of

4. Data processing

1) Inhibition (%) = [1- (D5 test object well reading-D0 reading mean)/(D5 solvent control well reading mean-D0 reading mean) ]x100%;

2) GI ₅₀ values were plotted and calculated based on inhibition.

Experimental results and conclusions

TABLE 4 in vitro cytological Activity of the Compounds of the invention

As can be seen from Table 4, the compounds of the present invention can effectively inhibit proliferation of T47D, MDA-MB-453 cells mutated with PI3K alpha H1047R.

Claims

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

Wherein,

each X ₄ is independently selected from-CR ^2aR^2b -;

y is selected from-CR ^3aR^3b-、-O-、-NR^3c -or-S-;

m, s, t are each independently selected from 0, 1,2, 3 or 4.

2. The compound of claim 1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein,

Ring B is selected from phenyl or 5-6 membered heteroaryl;

m, s, t are each independently selected from 0, 1,2 or 3.

3. The compound, pharmaceutically acceptable salt thereof, or stereoisomer thereof according to claim 1 or 2, wherein,

Ring a and the attached X ¹、X² together form a 5-6 membered monocycloalkyl, 5-6 membered monocycloheteroaryl, phenyl, 8-10 membered fused ring group, 8-10 membered fused heterocyclic group or 8-10 membered fused heteroaryl;

Each R ₁, each R ₂ is independently selected from halogen, hydroxy, nitro, cyano, amino, carboxy, C _1-4 alkyl, halo C _1-4 alkyl, hydroxy C _1-4 alkyl, amino C _1-4 alkyl, -NR ^aR^b、-OR^a、-C(O)OR^a, or the following optionally substituted with substituents: -L ₂ -3-6 membered cycloalkyl, -L ₂ -3-6 membered heterocyclyl, -L ₂ -5-6 membered heteroaryl, -L ₂ -phenyl; the substituent is selected from halogen, hydroxy, nitro, amino, cyano, carboxyl, C _1-4 alkyl, C _1-4 alkoxy or halogenated C _1-4 alkyl;

R ^a、R^b is independently at each occurrence selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, C _1-4 alkyl, halo C _1-4 alkyl, C _1-4 alkoxy, or halo C _1-4 alkoxy;

L ₂ is selected from -CH₂-、-CH₂-CH₂-、-CH₂-CH₂-CH₂-、-CH₂(CH₃)-CH₂-、-CH₂-O-、-CH₂-CH₂-O-、-O- or-NH-;

R^1a、R^1b、R^1c、R^2a、R^2b、R^3a、R^3b、R^3c、R^4a、R^4b、R^4c Each occurrence is independently selected from hydrogen, halogen, C _1-4 alkyl, halogenated C _1-4 alkyl, C _1-4 alkoxy, halogenated C _1-4 alkoxy.

4. The compound, pharmaceutically acceptable salt thereof, or stereoisomer thereof according to any one of claim 1 to 3, wherein,

When (when)X ₁、X₂ is selected from CH or N, X ₃ is selected from-CH ₂ -, -NH, -O-, -S-, -C (O) -, -S (O) -or-S (O) ₂ -, when representing a single bond;

5. The compound, pharmaceutically acceptable salt thereof, or stereoisomer thereof according to any one of claim 1 to 4, wherein,

Y is selected from-CHR ^3a -, -O-or-NR ^3c -;

l ₁ is selected from-CHR ^4a-CHR^4a-、-CHR^4a-NR^4c -or-CHR ^4a -O-;

Ring B is selected from phenyl or 6 membered heteroaryl;

m is selected from 0, 1 or 2;

s, t are each independently selected from 0, 1, 2 or 3.

6. The compound, pharmaceutically acceptable salt thereof, or stereoisomer thereof according to any one of claim 1 to 5, wherein,

Preferably, ring a and the X ₁、X₂ to which it is attached together constitute the following group:

ring B is selected from phenyl or 6 membered heteroaryl; preferably, ring B is selected from phenyl, pyridinyl or pyrimidinyl;

Each R ₁, each R ₂ is independently selected from halogen, hydroxy, nitro, cyano, amino, carboxy, C _1-4 alkyl, halo C _1-4 alkyl, hydroxy C _1-4 alkyl, amino C _1-4 alkyl, -NR ^aR^b、-OR^a、-C(O)OR^a; preferably, each R ₁, each R ₂ is independently selected from halogen, hydroxy, nitro, cyano, amino, carboxy, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, monofluoromethyl, difluoromethyl, trifluoromethyl, -NR ^aR^b、-OR^a、-C(O)OR^a;

R ₃、R₄、R₅ is independently selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, C _1-4 alkyl, halogenated C _1-4 alkyl, C _1-4 alkoxy, or halogenated C _1-4 alkoxy; preferably, R ₃、R₄、R₅ is each independently selected from hydrogen, halogen, hydroxy, amino, nitro, cyano, carboxy, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoromethoxy, difluoromethoxy or trifluoromethoxy;

R ^3a、R^3c、R^4a、R^4c、R^a、R^b is independently at each occurrence selected from hydrogen, halogen, C _1-4 alkyl, halogenated C _1-4 alkyl, C _1-4 alkoxy, or halogenated C _1-4 alkoxy; preferably, R ^3a、R^3c、R^4a、R^4c、R^a、R^b is independently selected at each occurrence from hydrogen, fluoro, chloro, bromo, iodo, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoromethoxy, difluoromethoxy or trifluoromethoxy;

m is selected from 0, 1 or 2;

s, t are each independently selected from 0, 1, 2 or 3.

7. The compound of claim 1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, selected from the group consisting of:

8. A pharmaceutical formulation comprising a compound according to any one of claims 1 to 7, a pharmaceutically acceptable salt or stereoisomer thereof, wherein the pharmaceutical formulation comprises one or more pharmaceutically acceptable excipients and is in any one of the pharmaceutically acceptable dosage forms.

9. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, characterized in that it comprises one or more second therapeutically active agents; optionally, the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers and/or diluents.

10. Use of a compound of any one of claims 1-7, a pharmaceutically acceptable salt or stereoisomer thereof, a pharmaceutical formulation of claim 8, or a pharmaceutical composition of claim 9 for the manufacture of a medicament for the treatment and/or prophylaxis of diseases and related conditions mediated by mutant pi3kα selected from cancer or benign tumors selected from lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, thyroid cancer, female genital tract cancer, lymphoma, neurofibromatosis, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, small cell lung cancer, non-small cell lung cancer, gastrointestinal stromal tumor, mast cell tumor, multiple myeloma, melanoma, leukemia, glioma or sarcoma; preferably, the mutant pi3kα has the E542K, E545K and H1047R mutations.