Fused ring compound, and preparation method therefor and use thereof

Abstract

Provided are a class of fused ring compounds, and a preparation method therefor and the use thereof. Specifically, provided are a compound of the following formula or a pharmaceutically acceptable form thereof, a pharmaceutical composition thereof, and a preparation method therefor and the use thereof. The compound can be used as a drug for treating diseases related to abnormal cell proliferation.

Description

Condensed ring compound and its preparation method and application Technical Field

[0001] The present invention relates to a class of condensed ring compounds, as well as their preparation methods and applications in the medical field. Background Art

[0002] Camptothecin (CPT) is a pentacyclic compound isolated from Camptotheca acuminata (see the structural formula below). Due to its excellent anticancer activity, it was introduced into clinical practice in the early 1970s. However, clinical trials were subsequently discontinued due to severe side effects such as vomiting, diarrhea, and hair loss.

[0003] Camptothecin and its derivatives have strong anti-tumor activity in animal models of ovarian, lung, colorectal, and breast cancer (Nature Review Cancer. 2006, 6, 789). Research data show that camptothecin can form a ternary complex with cellular DNA topoisomerase I, thereby inhibiting DNA unwinding, leading to DNA replication blockage and cell death (Cancer Res. 1989, 49, 6365).

[0004] Currently, several camptothecins have been approved for the treatment of tumors (Med. Res. Rev. 2015, 35, 753), for example, belotecan for the treatment of ovarian cancer and small cell lung cancer, topotecan for ovarian cancer, and irinotecan for the treatment of colorectal cancer.

[0005] Camptothecin drugs or derivatives often have hematotoxicity and gastrointestinal side effects. The hematotoxicity is caused by bone marrow suppression, which is manifested as thrombocytopenia, anemia, neutropenia, leukopenia, etc. Gastrointestinal adverse reactions include diarrhea, nausea, vomiting, etc. Clinical studies have found that it is possible to improve the safety and efficacy of camptothecin compounds by adjusting the activity of camptothecin compounds, improving the pharmacokinetic properties of camptothecin compounds, reducing the dosage, or forming antibody-drug conjugates with antibodies. Therefore, the research and development of camptothecin compounds and their conjugates with novel structures that can improve efficacy and improve safety issues still has a high clinical demand and application value. Summary of the Invention

[0006] The present invention provides a camptothecin compound with a novel structure. The camptothecin compound has good anti-tumor activity and good metabolic stability and is expected to be used in the treatment of tumor diseases.

[0007] In a first aspect, the present invention provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled compound, metabolite, and prodrug thereof, wherein the compound has the structure shown below:

[0008] in,

[0009] to exist or not to exist;

[0010] R1 is fluorine or chlorine,

[0011] R2 is selected from methyl, chloro, hydroxy or amino;

[0012] Alternatively, R1 and R2 together with the carbon atom to which they are attached form

[0013] R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring;

[0014] Ring A is selected from:

[0015] X is selected from oxygen or sulfur;

[0016] n is selected from 1 to 5;

[0017] when When the methyl group exists, R1 is fluorine, R2 is methyl, R3 is hydrogen, and X is oxygen, ring A is not morpholinyl.

[0018] In some embodiments, the compound has the structure shown below:

[0019] in,

[0020] R1 is fluorine or chlorine,

[0021] R2 is selected from methyl, chloro, hydroxy or amino;

[0022] Alternatively, R1 and R2 together with the carbon atom to which they are attached form

[0023] R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring;

[0024] Ring A is selected from: Preferred

[0025] X is selected from oxygen or sulfur;

[0026] n is selected from 1 to 5;

[0027] When R1 is fluorine, R2 is methyl, R3 is hydrogen, and X is oxygen, ring A is not morpholinyl.

[0028] In some embodiments, the compound has the structure shown below:

[0029] in,

[0030] R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring;

[0031] Ring A is selected from: Preferred

[0032] X is selected from oxygen or sulfur;

[0033] n is selected from 1 to 5;

[0034] When R1 is fluorine, R2 is methyl, R3 is hydrogen, and X is oxygen, ring A is not morpholinyl.

[0035] In some embodiments, the compound has the structure shown below:

[0036] in,

[0037] R1 is fluorine or chlorine,

[0038] R2 is selected from methyl, chloro, hydroxy or amino;

[0039] Alternatively, R1 and R2 together with the carbon atom to which they are attached form

[0040] R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring;

[0041] Ring A is selected from: Preferred

[0042] X is selected from oxygen or sulfur;

[0043] n is selected from 1-5.

[0044] In some embodiments, the compound has the structure shown below:

[0045] in,

[0046] R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring;

[0047] Ring A is selected from: Preferred

[0048] X is selected from oxygen or sulfur;

[0049] n is selected from 1-5.

[0050] In some embodiments, the compound has the structure shown below:

[0051] in,

[0052] R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring;

[0053] Ring A is selected from: Preferred

[0054] X is selected from oxygen or sulfur;

[0055] n is selected from 1-5.

[0056] In some embodiments, in the compounds of Formula I to Formula V above, X is selected from oxygen.

[0057] In some embodiments, in the compounds of Formula I to Formula V above, X is selected from sulfur.

[0058] In some embodiments, in the compounds of formula I to formula V, R3 is independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms and the connecting atom connect to form a 3-6 membered carbocyclic ring or a 3-6 membered heterocyclic ring.

[0059] In some embodiments, in the compounds of formula I to formula V, R3 is independently selected from hydrogen, C 1-4 Alkyl, halogen.

[0060] In some embodiments, in the compounds of Formula I to Formula V above, R3 is independently selected from hydrogen, methyl, ethyl, isopropyl, fluorine, chlorine, bromine or iodine.

[0061] In some embodiments, in the compounds of formula I to formula V above, ring A is selected from In some embodiments, in the compounds of formula I to formula V above, ring A is selected from

[0062] In some embodiments, in the compounds of formula I to formula V above, ring A is selected from

[0063] In some embodiments, in the compounds of formula I to formula V above, Selected from

[0064] In some embodiments, in the compounds of formula I to formula V above, Selected from

[0065] In some embodiments, in the compounds of formula I to formula V above, Selected from

[0066] In some embodiments, in the compounds of Formula I to Formula V above, n is selected from 1, 2, 3, 4, and 5.

[0067] In some embodiments, in the compounds of Formula I to Formula V above, n is selected from 1 and 2.

[0068] In some embodiments, the compound has the structure shown below:

[0069] In some embodiments, the compound has the structure shown below:

[0070] In some embodiments, the compound has the structure shown below:

[0071] Another aspect of the present invention provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite and prodrug thereof, wherein the compound has the structure shown below:

[0072] definition

[0073] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as those commonly understood by those skilled in the art. References to technology used herein are intended to refer to technology commonly understood in the art, including variations of technology or substitutions of equivalent technology that would be apparent to those skilled in the art. While it is believed that the following terms are well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

[0074] The terms "comprises," "comprising," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps.

[0075] As used herein, the "*" marked in a compound structural formula indicates that the marked carbon atom is a chiral carbon atom, and the present invention includes a pair of enantiomers formed by this chiral carbon atom. If a compound contains two different chiral carbon atoms, the present invention includes the four optical isomers formed by these chiral carbon atoms.

[0076] As used herein, Indicates the site where the structural fragment is connected to the rest of the molecule.

[0077] The term "alkyl" is defined as a straight or branched chain saturated aliphatic hydrocarbon group. In some embodiments, the alkyl group has 1 to 12, such as 1 to 6, carbon atoms. For example, as used herein, the term "C 1-6 "Alkyl" refers to a linear or branched group of 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl and n-hexyl), which is optionally substituted by 1 or more (such as 1, 2 or 3) suitable substituents.

[0078] The term "hydroxyalkyl" refers to an alkyl group substituted by one or more hydroxy groups, wherein the alkyl group is as defined above. For example, the term "C 1-6 "Hydroxyalkyl" refers to a hydroxyalkyl group having 1 to 6 carbon atoms. Common hydroxyalkyl groups include (but are not limited to) -CH2OH, -CH2CH2OH, -CH2CH(OH)2, and -(CH2)3OH.

[0079] The term "aminoalkyl" refers to an alkyl group substituted by one or more amino groups, wherein the alkyl group is as defined above. For example, the term "C 1-6 "Aminoalkyl" refers to an aminoalkyl group having 1 to 6 carbon atoms. Common aminoalkyl groups include (but are not limited to) -CH2-NH2, -CH2CH2-NH2, -CH2CH(NH2)2, and -(CH2)3-NH2.

[0080] The term "alkenyl" refers to a straight or branched chain hydrocarbon group containing at least one carbon-carbon double bond, including, for example, "C 2-6 Alkenyl", "C 2-4 Examples include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1,3-butadienyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1,4-hexadienyl, and the like.

[0081] The term "alkynyl" refers to a straight or branched chain hydrocarbon group containing at least one carbon-carbon triple bond. 2-6 Alkynyl", "C 4-6 Examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 1,3-butadiynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,4-hexadiynyl, and the like.

[0082] The term "cycloalkyl" refers to a saturated cyclic hydrocarbon group, including but not limited to monocyclic alkyl and bicyclic alkyl (such as spirocyclic alkyl, bicyclic alkyl and bridged cycloalkyl).3-6 "Cycloalkyl" refers to a cycloalkyl group having 3 to 6 ring carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc., which may be optionally substituted with 1 or more (such as 1, 2 or 3) suitable substituents, for example methyl substituted cyclopropyl.

[0083] The term "carbocyclyl" refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic structure, a hydrocarbon group connected through a ring carbon. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0084] The term "carbocycle" refers to a saturated or unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (for example, a monocyclic ring such as a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, a cycloheptane ring, a cyclooctane ring, a cyclononane ring, or a bicyclic ring, including a spirocyclic, fused or bridged system (such as a bicyclo[1.1.1]pentane ring, a bicyclo[2.2.1]heptane ring, a bicyclo[3.2.1]octane ring or a bicyclo[5.2.0]nonane ring, a decalin ring, etc.), which may be optionally substituted with one or more (such as one, two or three) suitable substituents. The term "3-6 membered carbocycle" refers to a carbocycle containing 3, 4, 5 or 6 ring-forming carbon atoms.

[0085] The term "heterocyclyl" or "heterocycle" refers to a saturated or partially saturated, monocyclic or polycyclic (such as a bicyclic) non-aromatic cyclic structure, the ring atoms of which are composed of carbon atoms and at least one (e.g., 1, 2, or 3) heteroatoms selected from nitrogen, oxygen, and sulfur. If the valence bond requirements are met, the heterocyclyl can be connected to the rest of the molecule via any one of the ring atoms. The heterocyclyl in the present invention is preferably a 3-6 membered heterocyclyl. The term "3-6 membered heterocyclyl" used in the present invention refers to a heterocyclyl having 3 to 6 ring atoms, including 3-membered heterocyclyl, 4-membered heterocyclyl, 5-membered heterocyclyl, and 6-membered heterocyclyl, including nitrogen-containing heterocyclyl, oxygen-containing heterocyclyl, such as 4-6 membered heterocyclyl, such as 4-6 membered nitrogen-containing heterocyclyl, 4-6 membered oxygen-containing heterocyclyl, 5-6 membered oxygen-containing heterocyclyl, and 5-membered oxygen-containing heterocyclyl. Common heterocyclic groups include, but are not limited to, azetidinyl, oxetanyl, tetrahydrofuryl, pyrrolidinyl, pyrrolidinonyl, imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, piperazinyl, and morpholinyl. The heterocyclic groups of the present invention may be optionally substituted with one or more substituents described herein. The heterocyclic groups of the present invention may be optionally fused to one or more aromatic or non-aromatic rings.

[0086] The term "oxygen-containing heterocycle" refers to a heterocycle as described above in which one or more (e.g., 1, 2, or 3) ring atoms are oxygen atoms, such as a 5-6 membered oxygen-containing heterocycle, a 5-membered oxygen-containing heterocycle, and specific examples include but are not limited to an oxirane ring, a tetrahydrofuran ring, a furan ring, a tetrahydropyran ring, a pyran ring, a 1,3-dioxolane ring, and the like.

[0087] The "nitrogen-containing heterocycle" described in the present invention refers to a heterocycle as described above in which one or more (eg, 1, 2 or 3) ring atoms are nitrogen atoms.

[0088] The term "haloalkyl" refers to an alkyl group substituted by one or more (such as 1, 2 or 3) the same or different halogen atoms, wherein alkyl is defined as above. For example, the term "C 1-6 "Haloalkyl" refers to a haloalkyl group having 1 to 6 carbon atoms. Common haloalkyl groups include (but are not limited to) -CH2F, -CHF2, -CF3, -CH2CF3, -CF2CF3, -CH2CH2CF3, -CH2Cl, etc. The haloalkyl group in the present invention is optionally substituted with one or more substituents described herein.

[0089] The term "alkoxy" refers to a group having the structure "alkyl-O-", wherein alkyl is as defined above. 1-6 Alkoxy, C 1-4 Alkoxy, C 1-3 Alkoxy or C 1-2 Alkoxy, etc. Common alkoxy groups include (but are not limited to) methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, etc. The alkoxy group in the present invention is optionally substituted with one or more substituents described herein.

[0090] The term "alkoxyalkyl" refers to an alkyl group substituted by one or more (e.g., 1, 2, 3, or 4) alkoxy groups, wherein alkoxy and alkyl are as defined above. For example, the term "C 1-6 "Alkoxyalkyl" refers to an alkyl group having 1-6 carbon atoms, substituted by one or more (e.g., 1, 2, 3, or 4) alkoxy groups. Common alkoxyalkyl groups include (but are not limited to) CH3O-CH2-, C2H5-O-CH2-, C2H5-O-CH2CH2-, etc.

[0091] The term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

[0092] The term "nitrogen oxide" refers to an oxide (eg, mono- or di-oxide) of at least one nitrogen atom in the structure of the present invention. Nitrogen mono-oxides may exist as a single positional isomer or a mixture of positional isomers.

[0093] If a substituent is described as being "optionally substituted," the substituent may be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more substituents from the list of substituents, one or more hydrogens on the carbon (to the extent of any hydrogens present) may be replaced, individually and / or collectively, with independently selected optional substituents. If a nitrogen of a substituent is described as being optionally substituted with one or more substituents from the list of substituents, one or more hydrogens on the nitrogen (to the extent of any hydrogens present) may each be replaced with an independently selected optional substituent.

[0094] If a functional group or structural moiety is described as "substituted or unsubstituted," the functional group or structural moiety may be (1) unsubstituted or (2) substituted.

[0095] The term "substituted" refers to the replacement of one or more (e.g., 1, 2, 3, 4, or 5) hydrogen atoms on a designated compound or structural fragment by a substituent, provided that the normal valence of the designated atom in the present case is not exceeded and the substitution forms a stable compound. Combinations of substituents and / or variables are permitted only if such combinations form stable compounds. For example, the substituents are each independently composed of one or more of the following structures: -O-, -S-, -NR'-, halogen, -CN, -OH, -NH2, -NO2, -CN, =O, C 1-6 (Ethylene) alkyl, C 1-6 Halogenated (ene) alkyl, C 1-6 Alkoxy, C 2-6 (Ethylene) alkenyl, C 2-6 (Ene) alkynyl, C 3-8 (Ethyl) cycloalkyl, 3-8 membered (ethyl) heterocyclic group, C 6-10 (E)aryl and 5-10 membered (E)heteroaryl, etc. If a substituent is described as being "independently selected" from a group of functional groups, each substituent is selected independently of the other. Thus, each substituent may be the same as or different from another (other) substituent.

[0096] As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5 or 10, where reasonable.

[0097] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0098] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In a compound with one or more (e.g., one, two, three, or four) asymmetric centers, it can produce a racemic mixture, a single enantiomer, a diastereomeric mixture, and a separate diastereomer. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. It is understood that the scope of this application encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%).

[0099] In this article, solid lines can be used Solid wedge or virtual wedge The carbon-carbon bonds of the compounds of the present invention are depicted. The use of solid lines to depict bonds to asymmetric carbon atoms is intended to indicate that all possible stereoisomers at that carbon atom are included (e.g., specific enantiomers, racemic mixtures, etc.). The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the indicated stereoisomers exist. When present in a racemic mixture, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the present invention are intended to exist as stereoisomers, including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, atropisomers, and mixtures thereof. The compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

[0100] The present invention encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs or mixtures of more than one polymorph in any ratio.

[0101] It should also be understood that certain compounds of the present invention may be used therapeutically in free form or, where appropriate, in the form of pharmaceutically acceptable derivatives thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, metabolites, or prodrugs that, upon administration to a patient in need thereof, are capable of directly or indirectly providing a compound of the present invention or a metabolite or residue thereof. Therefore, when reference is made herein to a "compound of the present invention," such various derivative forms of the compound are also intended to be encompassed.

[0102] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.

[0103] Suitable acid addition salts are formed from acids that form pharmaceutically acceptable salts and include aspartate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, and the like.

[0104] Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts, including aluminum salts, arginine salts, choline salts, diethylamine salts and the like.

[0105] For a review of suitable salts see Stahl and Wermuth, “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.

[0106] The term "ester" refers to esters derived from the compounds of the general formula herein, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the present invention in the form of free acid or alcohol). The compounds of the present invention themselves may also be esters.

[0107] The compounds of the present invention may exist in the form of solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0108] Also included within the scope of the present invention are metabolites of the compounds of the present invention, i.e., substances formed in vivo upon administration of the compounds of the present invention. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic hydrolysis, and the like of the administered compound. Thus, the present invention includes metabolites of the compounds of the present invention, including compounds produced by contacting a compound of the present invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

[0109] The present invention further includes within its scope prodrugs of the compounds of the present invention. Typically, such prodrugs will be functional group derivatives of the compounds that are readily converted into the desired therapeutically active compound in vivo. Therefore, in these cases, the term "administering" as used in the methods of treatment of the present invention should include treating various diseases or conditions with one or more prodrug forms of the claimed compounds, but the prodrug forms are converted into the aforementioned compounds in vivo after administration to the individual. For example, "Design of Prodrug", ed. H. Bundgaard, Elsevier, 1985, describes conventional methods for selecting and preparing suitable prodrug derivatives.

[0110] The present invention further includes within its scope isotopically labeled compounds of the invention which are identical to the compounds of the invention except that one or more atoms are replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature.

[0111] The present invention also encompasses compounds of the present invention that contain protecting groups. During any process for preparing the compounds of the present invention, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules involved, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved using conventional protecting groups, for example, those described in Protective Groups in Organic Chemistry, ed. JFW McOmie, Plenum Press, 1973; and TW Greene & P.GM Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

[0112] Pharmaceutical composition

[0113] In a third aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers.

[0114] The term "pharmaceutical composition" refers to a composition that can be used as a medicament, which comprises an active pharmaceutical ingredient (API) (or therapeutic agent) and, optionally, one or more pharmaceutically acceptable carriers. The term "pharmaceutically acceptable carrier" refers to an excipient that is administered with a therapeutic agent and that is suitable, within the scope of sound medical judgment, for contact with the tissues of humans and / or other animals without excessive toxicity, irritation, allergic reaction, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0115] The pharmaceutical compositions can act systemically and / or locally, which can be achieved by suitable dosage forms, including but not limited to tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, and syrups.

[0116] The pharmaceutical composition may contain 0.01 mg to 1000 mg of at least one compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof.

[0117] The present invention also provides a method for preparing the above-mentioned pharmaceutical composition or its corresponding formulation, which comprises combining at least one compound of the present invention or its pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug with one or more pharmaceutically acceptable carriers.

[0118] Medicine box products

[0119] In a fourth aspect, the present invention provides a kit product comprising:

[0120] a) at least one compound described herein as a first therapeutic agent, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled, metabolite or prodrug thereof, or a pharmaceutical composition as a first pharmaceutical composition;

[0121] b) optionally at least one additional therapeutic agent as a second therapeutic agent, or a pharmaceutical composition comprising an additional therapeutic agent as a second pharmaceutical composition; and

[0122] c) optional packaging and / or instructions.

[0123] The above-mentioned kit product may contain 0.01 mg to 1000 mg of at least one compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof.

[0124] The present invention also provides a method for preparing the above-mentioned drug kit, which comprises combining at least one compound of the present invention or its pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug or the above-mentioned pharmaceutical composition with at least one other therapeutic agent or pharmaceutical composition containing other therapeutic agents, packaging and / or instructions.

[0125] Medical uses

[0126] The compound of the present invention can exhibit a strong effect of inhibiting abnormal cell proliferation.

[0127] Therefore, the present application provides the compounds of the present invention or their pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope labels, metabolites and prodrugs, the above-mentioned pharmaceutical compositions or the above-mentioned drug kit products, which are used to treat diseases related to abnormal cell proliferation.

[0128] In addition, the present application also provides the use of the compound of the present invention or its pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled substances, metabolites and prodrugs, the above-mentioned pharmaceutical compositions or the above-mentioned drug kit products in the preparation of drugs for treating diseases related to abnormal cell proliferation.

[0129] In some embodiments, the disorder involving abnormal cell proliferation includes, but is not limited to, a tumor, such as an advanced solid tumor.

[0130] The present application also provides the use of a compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite and prodrug thereof or a pharmaceutical composition of the present invention in the preparation of a preparation for inhibiting the proliferation of tumor cells. In certain embodiments, the preparation is used for in vivo or in vitro administration. For example, the preparation can be administered to a subject to inhibit the proliferation of tumor cells in the subject; alternatively, the preparation can be administered to in vitro cells (e.g., cell lines or cells from a subject) to inhibit the proliferation of tumor cells in vitro.

[0131] The tumors described in the present invention include (but are not limited to): brain tumor, 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, female reproductive tract cancer, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, mast cell tumor, multiple myeloma, melanoma, glioma or sarcoma.

[0132] Treatment

[0133] In another aspect, the present invention provides a method for treating diseases involving abnormal cell proliferation, comprising the following steps: administering a therapeutically effective amount of a compound of the present invention or its pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope labels, metabolites and prodrugs, or the pharmaceutical composition or kit described above to an individual in need thereof.

[0134] The term "effective amount" refers to a dose that can induce a biological or medical response in cells, tissues, organs or organisms (eg, individuals) and is sufficient to achieve the desired preventive and / or therapeutic effect.

[0135] The dosage regimen can be adjusted to provide the optimal desired response. For example, the drug may be administered in a single dose, divided doses may be administered over time, or the dosage may be proportionally reduced or increased based on the actual situation. It will be understood that for any particular individual, the specific dosage regimen should be adjusted according to the needs and the professional judgment of the person administering the composition or supervising the administration of the composition.

[0136] The dosage of the compound of the present invention will depend on the individual situation, the severity of the disease or illness, the rate of administration, the disposal of the compound and the judgment of the prescribing physician. Generally speaking, the effective amount is about 0.001-10000mg / kg subject body weight / day. In appropriate cases, the effective amount is about 0.01-1000mg / kg subject body weight / day. About 0.01-1000mg / kg subject body weight, usually about 0.1-500mg / kg subject body weight, can be administered every day, every two days or every three days. An exemplary dosage regimen is once a day or more, or once a week or more, or once a month or more. During multiple administration, the intervals between single doses can generally be daily, weekly, monthly or annual. Alternatively, it can be administered in the form of a sustained-release formulation, in which case a lower frequency of administration is required. Dosage and frequency can vary according to the half-life of the drug in the subject, or can vary according to whether it is a preventive application or a therapeutic application. In preventive applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time; in therapeutic applications, a relatively high dosage is sometimes required at shorter intervals until the progression of the disease is slowed or stopped, preferably until the individual exhibits partial or complete improvement in disease symptoms, after which preventive applications can be employed.

[0137] The term "treat" refers to the alleviation or elimination of a targeted disease or condition. If a subject receives a therapeutic amount of a compound of the present invention, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention, and at least one indicator or symptom of the subject exhibits observable and / or detectable relief and / or improvement, the subject has been successfully "treated." It will be understood that treatment encompasses not only complete treatment but also less than complete treatment while achieving some biologically or medically relevant outcome.

[0138] The term "administrate / administrating / administration" (or "dosage") refers to the process of applying a pharmaceutically active ingredient (such as a compound of the present invention) or a pharmaceutical composition comprising a pharmaceutically active ingredient (such as a pharmaceutical composition of the present invention) to an individual or its cells, tissues, organs, biological fluids, and other parts, so as to bring the pharmaceutically active ingredient or pharmaceutical composition into contact with the individual or its cells, tissues, organs, biological fluids, and other parts. Common administration routes include (but are not limited to) oral administration, subcutaneous administration, intramuscular administration, subperitoneal administration, ocular administration, nasal administration, sublingual administration, rectal administration, vaginal administration, and the like.

[0139] The term "in need of" refers to a physician's or other health care provider's judgment that an individual needs or will benefit from a preventive and / or therapeutic procedure, which judgment is based on various factors within the physician's or other health care provider's area of ​​expertise.

[0140] The term "individual" (or subject) refers to a human or non-human animal. The individual of the present invention includes individuals (patients) suffering from a disease and / or condition and normal individuals. The non-human animals of the present invention include all vertebrates, such as non-mammals, such as birds, amphibians, reptiles, etc., and mammals, such as non-human primates, livestock and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

[0141] Preparation method

[0142] Another aspect of the present invention provides a method for synthesizing the compound.

[0143] Method A: The compound of formula (I) of the present invention can be synthesized by the following synthetic route:

[0144] Wherein, R1, R2, R3, X, A ring and n have the same meanings as described above, and PG is an amino protecting group;

[0145] Step 1

[0146] Compounds of formula (I)-SM1 and formula (I)-SM2 undergo condensation reaction to obtain a compound of formula (I)-IM1.

[0147] In some embodiments, this step is performed at a suitable temperature of 20-100°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0148] In some embodiments, this step is carried out in a suitable organic solvent, which can be selected from halogenated hydrocarbons (such as dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane (1,2-DCE), etc.), nitriles (such as acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), tetrahydrofuran (THF), 1,4-dioxane (Dioxane), dimethyl sulfoxide (DMSO) and any combination thereof, preferably DMF.

[0149] In some embodiments, this step is carried out in the presence of a suitable base, which includes an organic base or an inorganic base. The organic base can be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py); the inorganic base can be selected from potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), cesium carbonate (Cs2CO3) and NaOH, preferably DIPEA.

[0150] In some embodiments, this step is carried out in the presence of a suitable condensing agent, including HATU, HBTU, EDCI, DCC, DMTMM and HOBT, preferably DMTMM and HATU.

[0151] Step 2

[0152] The compound of formula (I)-IM1 is subjected to removal of the amino protecting group to obtain the compound of formula (I).

[0153] Method B: The compound of formula (III) in the present invention can be synthesized by the following synthetic route:

[0154] Wherein, R1, R2, R3, X, A ring and n have the same meanings as described above, and PG is an amino protecting group;

[0155] Step 1

[0156] Compounds of formula (III)-SM1 and formula (III)-SM2 undergo condensation reaction to obtain a compound of formula (III)-IM1.

[0157] In some embodiments, this step is performed at a suitable temperature of 20-100°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0158] In some embodiments, this step is carried out in a suitable organic solvent, which can be selected from halogenated hydrocarbons (such as dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane (1,2-DCE), etc.), nitriles (such as acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), tetrahydrofuran (THF), 1,4-dioxane (Dioxane), dimethyl sulfoxide (DMSO) and any combination thereof, preferably DMF.

[0159] In some embodiments, this step is carried out in the presence of a suitable base, which includes an organic base or an inorganic base. The organic base can be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py); the inorganic base can be selected from potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), cesium carbonate (Cs2CO3) and NaOH, preferably DIPEA.

[0160] In some embodiments, this step is carried out in the presence of a suitable condensing agent, including HATU, HBTU, EDCI, DCC, DMTMM and HOBT, preferably DMTMM and HATU.

[0161] Step 2

[0162] The compound of formula (III)-IM1 is subjected to removal of the amino protecting group to obtain the compound of formula (III).

[0163] Advantageous Effects of the Invention

[0164] The compounds of the present invention have good anti-tumor activity and can be used to treat abnormal cell proliferation disorders, including but not limited to advanced solid tumors. DETAILED DESCRIPTION

[0165] The present application is further described below by describing specific embodiments, but this is not intended to limit the present application. Those skilled in the art can make various modifications or improvements based on the teachings of the present application without departing from the basic idea and scope of the present application.

[0166] The abbreviations used in this invention have the following meanings:

[0167] The structures of the compounds described in the following examples were determined by nuclear magnetic resonance ( 1 H NMR) or mass spectrometry (MS).

[0168] Nuclear magnetic resonance (NMR)1 H NMR was measured using a Bruker 400 MHz nuclear magnetic resonance instrument; hexadeuterated dimethyl sulfoxide (DMSO-d6); and tetramethylsilane (TMS) as the internal standard.

[0169] The abbreviations used in the nuclear magnetic resonance (NMR) spectra in the examples are shown below.

[0170] s: singlet, d: doublet, t: triplet, q: quartet, m: multiplet, br: broad, J: coupling constant, Hz: hertz, DMSO-d6: deuterated dimethyl sulfoxide. δ values ​​are expressed in ppm.

[0171] The mass spectrometry (MS) was performed using an Agilent (ESI) mass spectrometer, model Agilent 6120B.

[0172] Example 1: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-4-oxoazetidine-2-carboxamide (Compound 1-5a)

[0173] The methanesulfonate salt of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizine[1,2-b]quinoline-10,13-dione (24 mg, 45.15 μmol) and (R)-4-oxoazetidine-2-carboxylic acid (6.00 mg, 52.13 μmol) were dissolved in DMF (1 mL). DIPEA (16 mg, 123.80 μmol) was added dropwise, causing the mixture to turn dark. HATU (20.59 mg, 54.18 μmol) was added, and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was directly purified by HPLC and freeze-dried to obtain the title compound (22 mg, 40.49 μmol).

[0174] Its structural characterization data are as follows:

[0175] MS m / z(ESI):533.3[M+H] +

[0176] 1H NMR(400MHz,DMSO)δ8.75(d,J=8.4Hz,1H),8.12(s,1H),7.82(d,J=10.8Hz,1H),7 .31(s,1H),6.54(s,1H),5.65-5.60(m,1H),5.43(s,2H),5.33-5.28(m,1H),5.15- 5.11(m,1H),4.00-3.98(m,1H),3.23-3.09(m,3H),2.92-2.87(m,1H),2.41(s,3H ),2.24-2.19(m,1H),2.14-2.09(m,1H),1.91-1.80(m,2H),0.87(t,J=7.2Hz,3H).

[0177] The preparation method is as follows:

[0178] Chromatographic column: Waters Sunfire Prep C18 OBD (5μm*19mm*150mm)

[0179] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0180] Example 2: Preparation of (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-4-oxoazetidine-2-carboxamide (Compound 1-5b)

[0181] (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizine[1,2-b]quinoline-10,13-dione methanesulfonate (38 mg, 71.49 μmol) was added to DMF (2 mL), followed by DIPEA (28.63 mg, 221.56 μmol). The mixture was stirred at 22°C for 20 minutes. Finally, (S)-4-oxoazetidine-2-carboxylic acid (12.75 mg, 110.78 μmol) was added, and the mixture was stirred at 22°C for 1 hour. The reaction mixture was purified by HPLC and freeze-dried to obtain the title compound (25 mg, 46.01 μmol).

[0182] Its structural characterization data are as follows:

[0183] MS m / z(ESI):533.3[M+H] +

[0184] 1H NMR (400MHz, DMSO) δ8.79(d,J=8.7Hz,1H),8.16(s,1H),7.81(d,J=11.0Hz,1H),7.30(s ,1H),6.54(s,1H),5.65-5.58(m,1H),5.43(s,2H),5.24(d,J=18.8Hz,1H),5.11(d,J=18 .8Hz,1H),4.02(dd,J=5.4,2.4Hz,1H),3.19(s,2H),3.13-3.06(m,1H),2.90(dd,J=10. 5,4.0Hz,1H),2.40(s,3H),2.24-2.10(m,2H),1.93-1.78(m,2H),0.87(t,J=7.3Hz,3H).

[0185] The preparation method is as follows:

[0186] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0187] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0188] Example 3: Preparation of N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3,3-difluoroazetidine-2-carboxamide (Compound 1-6)

[0189] Step 1: Preparation of tert-butyl 2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)-3,3-difluoroazetidine-1-carboxylate (Compound 1-6-1)

[0190] Methanesulfonate of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizine[1,2-b]quinoline-10,13-dione (50 mg, 94.06 μmol), 1-tert-butoxycarbonyl-3,3-difluoro-azetidine-2-carboxylic acid (24.54 mg, 103.47 μmol), and HATU (53.62 mg, 141.10 μmol) were dissolved in DMF (1 mL), and DIPEA (36.47 mg, 282.19 μmol) was added dropwise. The mixture was stirred for 1 hour. Water was added, and 1N hydrochloric acid was added dropwise to acidify the mixture. Ethyl acetate was added for extraction. The organic phase was washed with saturated brine, dried and concentrated to obtain a crude product (60 mg, 91.65 μmol), which was directly used for the next reaction.

[0191] Its structural characterization data are as follows:

[0192] MS m / z(ESI):516.2[M+H] +

[0193] Step 2: Preparation of N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3,3-difluoroazetidine-2-carboxamide (Compound 1-6)

[0194] Dissolve tert-butyl 2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)-3,3-difluoroazetidine-1-carboxylate (60 mg, 91.65 μmol, crude) in dioxane hydrochloride (1 mL) and stir for 1 hour. The mixture was concentrated under reduced pressure, purified by HPLC, and freeze-dried to give the title compound (16.10 mg, 25.88 μmol).

[0195] Its structural characterization data are as follows:

[0196] MS m / z(ESI):555.5[M+H] +

[0197] 1H NMR(400MHz,DMSO)δ8.79(ddd,J=21.1,15.8,8.9Hz,1H),7.80(d,J=11.0Hz,1H),7 .31(t,J=2.0Hz,1H),6.53(d,J=2.1Hz,1H),5.65-5.55(m,1H),5.43(s,2H),5.39-5 .05(m,3H),4.68-4.55(m,1H),3.92(d,J=8.1Hz,2H),3.23-3.08(m,2H),2.39(d,J= 5.1Hz,3H),2.21-2.09(m,2H),1.86(dt,J=15.0,7.4Hz,2H),0.87(t,J=7.3Hz,3H).

[0198] The preparation method is as follows:

[0199] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0200] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0201] Example 4: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)isoxazolidine-3-carboxamide (Compound 2-1a)

[0202] Step 1: Preparation of (R)-3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)isoxazolidine-2-carboxylic acid tert-butyl ester (Compound 2-1a-1)

[0203] The methanesulfonate salt of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizine[1,2-b]quinoline-10,13-dione (30 mg, 56.44 μmol) and (3R)-2-tert-butoxycarbonylisoxazolidine-3-carboxylic acid (13.49 mg, 62.08 μmol) were dissolved in DMF (1 mL). HATU (32.17 mg, 84.66 μmol) was added, and DIPEA (21.88 mg, 169.32 μmol) was added dropwise. The mixture was stirred for 1 hour. Water was added, 1N hydrochloric acid was added dropwise to acidify, and ethyl acetate was added for extraction. The organic phase was washed with saturated brine, dried and concentrated to obtain a crude product (35 mg, 55.15 μmol), which was directly used for the next reaction.

[0204] Its structural characterization data are as follows:

[0205] MS m / z(ESI):635.8[M+H] +

[0206] Step 2: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)isoxazolidine-3-carboxamide (Compound 2-1a)

[0207] Dissolve (R)-tert-butyl 3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)isoxazolidine-2-carboxylate (35 mg, 55.15 μmol, crude) in dioxane hydrochloride (1 mL) and stir for 1 hour. The reaction mixture was concentrated under reduced pressure, purified by HPLC, and freeze-dried to give the title compound (17.23 mg, 29.57 μmol).

[0208] Its structural characterization data are as follows:

[0209] MS m / z(ESI):535.1[M+H] +

[0210] 1H NMR (400MHz, DMSO) δ8.65(d,J=9.8Hz,1H),7.82-7.75(m,1H),7.31(d,J=2.7Hz,1H),6.52(d,J=7.7Hz,1H),5.58(s,2H),5.39(d,J=23.0Hz,2H),5. 36(s,1H),5.19(s,1H),4.06(s,2H),3.51(s,2H),3.29-3.20(m,2H),2.3 9(s,3H),2.18(d,J=5.7Hz,2H),1.90-1.80(m,2H),0.87(t,J=7.4Hz,3H).

[0211] The preparation method is as follows:

[0212] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0213] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0214] Example 5: Preparation of (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)isoxazolidine-3-carboxamide (Compound 2-1b)

[0215] Step 1: Preparation of (S)-3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)isoxazolidine-2-carboxylic acid tert-butyl ester (Compound 2-1b-1)

[0216] Methanesulfonate of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizine[1,2-b]quinoline-10,13-dione (30 mg, 56.4 μmol) and (S)-2-(tert-butoxycarbonyl)isoxazolidine-3-carboxylic acid (22.3 mg, 103 μmol) were dissolved in DMF (2 mL). DIPEA (26.6 mg, 205.9 μmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (39.1 mg, 102.9 μmol) were added. The mixture was stirred at 22°C for 1 hour. The reaction solution was purified by HPLC and freeze-dried to obtain the title compound (30.5 mg, 48.0 μmol).

[0217] Its structural characterization data are as follows:

[0218] MS m / z(ESI):637.2[M+H] +

[0219] Step 2: Preparation of (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)isoxazolidine-3-carboxamide (Compound 2-1b)

[0220] (S)-tert-Butyl 3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)isoxazolidine-2-carboxylate (30.5 mg, 48.0 μmol) was added to a single-necked vial. DCM (3 mL) was then added, followed by 1 mL of trifluoroacetic acid. The mixture was stirred at 22°C for 1 hour. The crude product was concentrated under reduced pressure at room temperature, purified by HPLC, and freeze-dried to afford the title compound (18.94 mg, 35.3 μmol).

[0221] Its structural characterization data are as follows:

[0222] MS m / z(ESI):535.2[M+H] +

[0223] 1H NMR (400MHz, DMSO) δ8.67(d,J=7.8Hz,1H),7.79(d,J=11.0Hz,1H),7.30(s,1H),5.58-5.51(m,1H),5.42(s,2H),5.22-5.17(m,2H),4 .13-3.85(m,3H),3.71-3.66(m,2H),3.16(t,J=6.0Hz,2H),2.39(s,3H),2.23-2.08(m,2H),1.91-1.79(m,2H),0.87(t,J=7.2Hz,3H).

[0224] The preparation method is as follows:

[0225] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0226] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% trifluoroacetic acid)

[0227] Example 6: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)thiazolidine-2-carboxamide (Compound 2-6a)

[0228] Step 1: Preparation of (R)-2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (Compound 2-6a-1)

[0229] The methanesulfonate of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (120 mg, 225.75 μmol mmol), (R)-3-(tert-butoxycarbonyl)thiazolidine-2-carboxylic acid (63 mg, 0.27 mmol) and DIPEA (88 mg, 0.68 mmol) were added to DMF (3 mL) and stirred for 5 minutes. HATU (129 mg, 338.63 μmol) was added and stirred at 25 ° C for 1 hour. The reaction solution was directly purified by C18 reverse column (ACN / H2O = 10-90%, 0.05% formic acid aqueous solution), and ethyl acetate (50 ml) was added to the purified solution and stirred for 5 minutes. The upper organic phase was separated and concentrated to obtain the title compound (120 mg, 0.18 mmol).

[0230] Its structural characterization data are as follows:

[0231] MS m / z(ESI):651.3[M+H] +

[0232] Step 2: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)thiazolidine-2-carboxamide (Compound 2-6a)

[0233] (R)-2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (120 mg, 0.18 mmol) was dissolved in 1,4-dioxane (4 mL), methanol (1 mL) and 1% CO 2 HCl. mL) mixed solvent, added hydrogen chloride-1,4-dioxane solution (4M, 1.4 mL), heated to 35 ° C for 3 hours, added methyl tert-butyl ether (5 ml), stirred and cooled to room temperature, allowed to stand, the supernatant was decanted, methyl tert-butyl ether (5 ml) was added, stirred for 15 minutes, allowed to stand, the supernatant was decanted, vacuum dried, acetonitrile (5 ml) and deionized water (5 ml) were added to dissolve, and lyophilized to obtain the title compound (85 mg, 0.14 mmol, hydrochloride).

[0234] Its structural characterization data are as follows:

[0235] MS m / z(ESI):551.2[M+H] +

[0236] Example 7: Preparation of (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)thiazolidine-2-carboxamide (Compound 2-6b)

[0237] Step 1: Preparation of (S)-2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (Compound 2-6b-1)

[0238] (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione methanesulfonate (120 mg, 225.75 μmol) and (2S)-3-tert-butoxycarbonylthiazolidine-2-carboxylic acid (63.20 mg, 270.90 μmol) were added to DMF (5 mL), followed by the addition of DIPEA (87.53 mg, 677.25 μmol) and HATU (129 mg, 338.63 μmol). After the addition was complete, the reaction was stirred at 22°C for 1 hour. The reaction solution was purified by HPLC and freeze-dried to obtain the title compound (120 mg, 184.41 μmol).

[0239] Its structural characterization data are as follows:

[0240] MS m / z(ESI):651.3[M+H] +

[0241] The preparation method is as follows:

[0242] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0243] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0244] Step 2: Preparation of (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)thiazolidine-2-carboxamide (Compound 2-6b)

[0245] (S)-tert-Butyl 2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)thiazolidine-3-carboxylate (120 mg, 184.41 μmol) was added to 1,4-dioxane (5 mL), followed by a 4 M hydrogen chloride-1,4-dioxane solution (2 mL). After addition, the mixture was stirred at 35°C for 2 hours. The reaction solution was purified by HPLC and freeze-dried to obtain the title compound (52 mg, 84.15 μmol, hydrochloride salt).

[0246] Its structural characterization data are as follows:

[0247] MS m / z(ESI):551.2[M+H] +

[0248] The preparation method is as follows:

[0249] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0250] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0251] Example 8: Preparation of (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)thiazolidine-4-carboxamide (Compound 2-9a)

[0252] Step 1: Preparation of (S)-4-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (Compound 2-9a-1)

[0253] The methanesulfonate of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizine[1,2-b]quinoline-10,13-dione (80 mg, 150.50 μmol), (S)-3-(tert-butyloxycarbonyl)thiazolidine-4-carboxylic acid (70.22 mg, 301.01 μmol), HATU (171.68 mg, 451.5 μmol) and DIPEA (97.25 mg, 752.51 μmol) were added to a DMF (6 mL) reaction system and stirred at 25°C for 18 hours. The reaction solution was prepared by HPLC and freeze-dried to obtain the title compound (85 mg, 130.63 μmol).

[0254] Its structural characterization data are as follows:

[0255] MS m / z(ESI):651.3[M+H] +

[0256] The preparation method is as follows:

[0257] Chromatographic column: Waters Xbridge Prep C18 OBD (5μm*19mm*150mm)

[0258] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0259] Step 2: Preparation of (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)thiazolidine-4-carboxamide (Compound 2-9a)

[0260] Dissolve (S)-tert-Butyl 4-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)thiazolidine-3-carboxylate (60 mg, 92.21 μmol) in HCl / 1,4-dioxane (5 mL) and stir at 25°C for 1 hour. The reaction mixture was concentrated to obtain the crude product, which was purified by preparative HPLC and freeze-dried to obtain the title compound (25 mg, 42.16 μmol).

[0261] Its structural characterization data are as follows:

[0262] MS m / z(ESI):551.2[M+H] +

[0263] 1 H NMR(400MHz,DMSO)δ8.65(d,J=8.4Hz,1H),7.77(d,J=10.4Hz,1H),7.29(s,1H), 6.53(s,1H),5.60-5.50(m,1H),5.42(s,2H),5.28-5.08(m,2H),4.14(d,J=8.4H z,1H),4.00(d,J=7.6Hz,1H),3.90-3.80(m,1H),3.20-3.10(m,2H),3.08-2.90( m,2H),2.38(s,3H),2.27-2.05(m,2H),1.95-1.75(m,2H),0.87(t,J=6.0Hz,3H).

[0264] The preparation method is as follows:

[0265] Chromatographic column: Waters Xbridge Prep C18 OBD (5μm*19mm*150mm)

[0266] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0267] Example 9: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)thiazolidine-4-carboxamide (Compound 2-9b)

[0268] Step 1: Preparation of (R)-4-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (Compound 2-9b-1)

[0269] (4R)-3-tert-Butyloxycarbonylthiazolidine-4-carboxylic acid (87.8 mg, 0.37 mmol), (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione methanesulfonate (200 mg, 0.37 mmol), and N,N-diisopropylethylamine (194.5 mg, 1.5 mmol) were dissolved in DMF (5 mL). HATU (171.7 mg, 0.45 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion, the reaction mixture was purified by reverse phase column chromatography to yield the title compound (70 mg, 0.11 mmol).

[0270] Its structural characterization data are as follows:

[0271] MS m / z(ESI):651.3[M+H] +

[0272] Step 2: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)thiazolidine-4-carboxamide (Compound 2-9b)

[0273] Dissolve (R)-tert-butyl 4-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)thiazolidine-3-carboxylate (70 mg, 0.11 mmol) in 1,4-dioxane (4 mL). Cool the mixture to 0°C and add a 4 M solution of hydrogen chloride in 1,4-dioxane (2 mL). The reaction mixture was allowed to react at room temperature for 1 hour. After completion of the reaction, the solvent was concentrated to obtain a crude product. This crude product was reacted to yield the title compound (33.06 mg, 58.84 μmol).

[0274] Its structural characterization data are as follows:

[0275] MS m / z(ESI):551.3[M+H] +

[0276] 1 H NMR (400MHz, DMSO) δ8.78-8.68(m,1H),7.80(d,J=10.8Hz,1H),7.31(s,1H),6.54( s,1H),5.60-5.54(m,1H),5.43(s,2H),5.30-5.14(m,2H),4.25-4.13(m,1H),4.07 -3.98(m,1H),3.86-3.77(m,1H),3.21-3.13(m,2H),3.09-3.01(m,1H),2.96-2.92 (m,1H),2.40(s,2H),2.24-2.09(m,2H),1.90-1.82(m,2H),0.87(t,J=7.2Hz,3H).

[0277] The preparation method is as follows:

[0278] Chromatographic column: Agilent Prep C18OBD (5μm*19mm*150mm)

[0279] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0280] Example 10: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-2,2-dimethylthiazolidine-4-carboxamide (Compound 2-10b)

[0281] Step 1: Preparation of (9H-fluoren-9-yl)methyl (R)-4-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)-2,2-dimethylthiazolidine-3-carboxylate (Compound 2-10b-1)

[0282] The methanesulfonate of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (85 mg, 159.91 μmol), (R)-3-(((9H-fluoren-9-yl)methoxy)carbonyl)-2,2- Dimethylthiazolidine-4-carboxylic acid (122.64 mg, 319.82 μmol), HATU (121.61 mg, 319.82 μmol) and DIPEA (72.33 mg, 559.68 μmol) were added to the DMF (5 mL) reaction system and stirred at 25 °C for 1 hour. The reaction solution was prepared by HPLC and freeze-dried to obtain the title compound (73 mg, 128.07 μmol).

[0283] Its structural characterization data are as follows:

[0284] MS m / z(ESI):801.1[M+H ]+

[0285] The preparation method is as follows:

[0286] Chromatographic column: Waters Xbridge Prep C18 OBD (5μm*19mm*150mm)

[0287] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0288] Step 2: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-2,2-dimethylthiazolidine-4-carboxamide (Compound 2-10b)

[0289] (9H-Fluoren-9-yl)methyl (R)-4-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)-2,2-dimethylthiazolidine-3-carboxylate (30 mg, 30.76 μmol) was dissolved in DMF (5 mL). Diethylamine (13.70 mg, 187.29 μmol) was added and stirred at 25°C for 1 hour. The reaction mixture was directly purified by preparative HPLC and freeze-dried to obtain the title compound (2 mg, 3.39 μmol).

[0290] Its structural characterization data are as follows:

[0291] MS m / z(ESI):579.1[M+H] +

[0292] 1 H NMR (400MHz, DMSO) δ8.84(d,J=8.4Hz,1H),7.82(d,J=10.8Hz,1H),7.31(s, 1H),6.54(s,1H),5.67-5.58(m,1H),5.42(s,2H),5.28-5.10(m,2H),4.03- 3.93(m,1H),3.24-3.16(m,3H),3.08-2.98(m,2H),2.41(s,3H),2.22-2.13 (m,2H),1.92-1.80(m,2H),1.62(s,3H),1.44(s,3H),0.87(t,J=7.2Hz,3H).

[0293] The preparation method is as follows:

[0294] Chromatographic column: Waters Xbridge Prep C18 OBD (5μm*19mm*150mm)

[0295] Mobile phase A: acetonitrile; mobile phase B: water

[0296] Example 11: Preparation of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizine[1,2-b]quinoline-10,13-dione (Compound 2-12)

[0297] Methanesulfonate of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizine[1,2-b]quinoline-10,13-dione (55 mg, 103.47 μmol), 1,1-dioxoisothiazolidine-3-carboxylic acid (34.18 mg, 206.94 μmol), HATU (78.69 mg, 206.94 μmol) and DIPEA (66.86 mg, 517.35 μmol) were added to a DMF (5 mL) reaction system and stirred at 25°C for 1 hour. The reaction solution was prepared by HPLC and freeze-dried to obtain the title compound (14 mg, 23.79 μmol).

[0298] Its structural characterization data are as follows:

[0299] MS m / z(ESI):583.2[M+H] +

[0300] 1 H NMR (400MHz, DMSO) δ8.62 (d, J = 8.4Hz, 1H), 7.80 (d, J = 10.8Hz, 1H), 7.35-7. 25(m,2H),6.53(br,1H),5.64-5.52(m,1H),5.43(s,2H),5.32-5.15(m,2H) ,4.10-4.01(m,1H),3.36-3.00(m,5H),2.71-2.63(m,1H),2.40(s,3H),2.3 1-2.21(m,1H),2.15-2.05(m,1H),1.93-1.80(m,2H),0.87(t,J=7.2Hz,3H).

[0301] The preparation method is as follows:

[0302] Chromatographic column: Waters Xbridge Prep C18 OBD (5μm*19mm*150mm)

[0303] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0304] Example 12: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)-4-oxazetidine-2-carboxamide (Compound 1'-5a)

[0305] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (33.33 mg, 81.4 μmol) and (R)-4-oxoazetidine-2-carboxylic acid (9.74 mg, 84.61 μmol) were dissolved in DMF (1 mL). DIPEA (16 mg, 123.80 μmol) was added dropwise, followed by PyBOP (45.76 mg, 87.93 μmol), and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was directly purified by HPLC and freeze-dried to obtain the title compound (9.11 mg, 17.72 μmol).

[0306] Its structural characterization data are as follows:

[0307] MS m / z(ESI):507.0[M+H] +

[0308] 1 H NMR (400MHz, DMSO) δ9.01-8.98(m,1H),8.36(d,J=8.4Hz,1H),8.12(s,1H),7.91(d,J=10.4Hz,1H),7.32(s,1H),6.53(s,1H),5.46-5.44(m, 3H),4.88(d,J=5.6Hz,2H),3.98-3.96(m,1H),3.13-3.08(m,2H),2.74-2.70(m,1H),2.49(s,3H),1.92-1.81(m,2H),0.87(t,J=7.2Hz,3H).

[0309] The preparation method is as follows:

[0310] Chromatographic column: Waters Sunfire Prep C18 OBD (5μm*19mm*150mm)

[0311] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% ammonium bicarbonate)

[0312] Example 13: Preparation of (S)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)-4-oxoazetidine-2-carboxamide (Compound 1'-5b)

[0313] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (30 mg, 73.28 μmol) was added to DMF (2 mL), followed by DIPEA (28.41 mg, 219.83 μmol). The mixture was stirred at 22°C for 20 minutes. Finally, (S)-4-oxoazetidine-2-carboxylic acid (12.65 mg, 109.91 μmol) was added, and the mixture was stirred at 22°C for 1 hour. The reaction solution was purified by HPLC and freeze-dried to obtain the title compound (8 mg, 15.01 μmol).

[0314] Its structural characterization data are as follows:

[0315] MS m / z(ESI):507.3[M+H] +

[0316] 1 H NMR (400MHz, DMSO) δ9.00(t,J=5.8Hz,1H),8.36(d,J=8.1Hz,1H),8.13(s,1H),7.91(d,J=10.8Hz,1H),7.32(s,1H),5.45(d,J=3.9Hz,4H),4.94- 4.85(m,2H),3.96(dd,J=5.5,2.4Hz,1H),3.14-3.07(m,1H),2.69(dd,J= 15.4,8.6Hz,1H),1.93-1.79(m,2H),1.23(s,2H),0.87(t,J=7.3Hz,3H).

[0317] The preparation method is as follows:

[0318] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0319] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0320] Example 14: Preparation of N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizine[1,2-b]quinolin-11-yl)methyl)-3,3-difluoroazetidine-2-carboxamide (Compound 1'-7)

[0321] Step 1: Preparation of tert-butyl 2-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyranyl[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)-3,3-difluoroazetidine-1-carboxylate (Compound 1'-7-1)

[0322] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (30 mg, 73.28 μmol), 1-tert-butoxycarbonyl-3,3-difluoro-azetidine-2-carboxylic acid (19.12 mg, 80.60 μmol), and HATU (41.77 mg, 109.91 μmol) were dissolved in DMF (1 mL). DIPEA (28.41 mg, 219.83 μmol) was added dropwise and stirred for 1 hour. Water was added, and 1N aqueous hydrochloric acid was added dropwise to acidify the mixture. Ethyl acetate was added for extraction. The organic phase was washed with saturated brine, dried, and concentrated to obtain a crude product (46 mg, 73.18 μmol), which was directly used for the next step.

[0323] Its structural characterization data are as follows:

[0324] MS m / z(ESI):629.3[M+H]+

[0325] Step 2: Preparation of N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizine[1,2-b]quinolin-11-yl)methyl)-3,3-difluoroazetidine-2-carboxamide (Compound 1'-7)

[0326] Dissolve tert-butyl 2-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyranyl[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)-3,3-difluoroazetidine-1-carboxylate (46 mg, 73.18 μmol, crude) in dioxane hydrochloride (1 mL) and stir for 1 hour. The reaction mixture was concentrated under reduced pressure, purified by HPLC, and freeze-dried to give the title compound (3.15 mg, 5.30 μmol).

[0327] Its structural characterization data are as follows:

[0328] MS m / z(ESI):529.5[M+H] +

[0329] 1 H NMR (400MHz, DMSO) δ9.02(s,1H),8.41(d,J=8.5Hz,1H),7.90(d,J=10.8Hz,1H),7.32(s,1H),6.53(s,1H),5.50(s,2H),5.44(s,2H),4.98(dd,J=14 .9,6.6Hz,1H),4.88-4.81(m,1H),4.54(s,1H),3.92(s,1H),3.62(s,1H), 3.38(s,1H),2.48-2.39(m,3H),1.92-1.81(m,2H),0.87(t,J=7.3Hz,3H).

[0330] The preparation method is as follows:

[0331] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0332] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0333] Example 15: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)isoxazolidine-3-carboxamide (Compound 2'-1a)

[0334] Step 1: Preparation of (R)-3-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)isoxazolidine-2-carboxylic acid tert-butyl ester (Compound 2'-1a-1)

[0335] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (30 mg, 73.28 μmol), (3R)-2-tert-butoxycarbonylisoxazolidine-3-carboxylic acid (17.51 ​​mg, 80.60 μmol), and HATU (32.17 mg, 84.66 μmol) were dissolved in DMF (1 mL). DIPEA (28.41 mg, 219.83 μmol) was added dropwise and stirred for 1 hour. Water was added, and 1N hydrochloric acid was added dropwise to acidify the mixture. Ethyl acetate was added for extraction. The organic phase was washed with saturated brine, dried, and concentrated to obtain a crude product (44 mg, 72.30 μmol), which was directly used for the next step.

[0336] Its structural characterization data are as follows:

[0337] MS m / z(ESI):609.8[M+H] +

[0338] Step 2: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)isoxazolidine-3-carboxamide (Compound 2'-1a)

[0339] Dissolve (R)-tert-butyl 3-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)isoxazolidine-2-carboxylate (44 mg, 72.30 μmol, crude) in dioxane hydrochloride (1 mL) and stir for 1 hour. The reaction mixture was concentrated under reduced pressure, purified by HPLC, and freeze-dried to obtain the title compound (6.40 mg, 11.16 μmol).

[0340] Its structural characterization data are as follows:

[0341] MS m / z(ESI):509.1[M+H] +

[0342] 1H NMR (400MHz, DMSO) δ8.85(s,1H),8.42(d,J=7.9Hz,1H),7.90(d,J=10.8Hz,1H),7.31(s,1H),6.52(s,1H),5.44(d,J=2.9Hz,4H),4.86 (ddd,J=21.0,15.5,6.2Hz,3H),3.90(s,2H),2.49-2.49(m,3H),2.33(dd,J=6.8,5.0Hz,3H),1.91-1.81(m,2H),0.87(t,J=7.3Hz,3H).

[0343] The preparation method is as follows:

[0344] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0345] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0346] Example 16: Preparation of (S)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)isoxazolidine-3-carboxamide (Compound 2'-1b)

[0347] Step 1: Preparation of (S)-3-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)isoxazolidine-2-carboxylic acid tert-butyl ester (Compound 2'-1b-1)

[0348] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizine[1,2-b]quinoline-3,14(4H)-dione (30 mg, 73.3 μmol) and (S)-2-(tert-butoxycarbonyl)isoxazolidine-3-carboxylic acid (23.7 mg, 10.9 μmol) were dissolved in DMF. To the 1% acetic acid methyl ester (2 mL) was added DIPEA (28.2 mg, 218.9 μmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (41.8 mg, 109.4 μmol). After the addition was complete, the mixture was stirred at 22°C for 1 hour. The reaction solution was purified by HPLC and freeze-dried to obtain the title compound (28.9 mg, 47.4 μmol).

[0349] Its structural characterization data are as follows:

[0350] MS m / z(ESI):609.2[M+H] +

[0351] Step 2: Preparation of (S)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)isoxazolidine-3-carboxamide (Compound 2'-1b)

[0352] (S)-tert-Butyl 3-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)isoxazolidine-2-carboxylate (28.9 mg, 47.4 μmol) was added to a single-necked vial. DCM (3 mL) was then added, followed by 1 mL of trifluoroacetic acid. The mixture was stirred at 22°C for 1 hour. The crude product was concentrated under reduced pressure at room temperature, purified by HPLC, and freeze-dried to obtain the title compound (13.88 mg, 27.2 μmol).

[0353] Its structural characterization data are as follows:

[0354] MS m / z(ESI):509.2[M+H] +

[0355] 1H NMR (400MHz, DMSO) δ8.92(s,1H),8.40(d,J=8.2Hz,1H),7.89(d,J=10.8Hz,1H),7.31(s,1H),5.51-5.46(m,4H), 4.88-4.61(m,2H),4.01-3.71(m,3H),2.42(s,3H),2.40-2.18(m,3H),1.94-1.78(m,2H),0.88(t,J=7.2Hz,3H).

[0356] The preparation method is as follows:

[0357] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0358] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% trifluoroacetic acid)

[0359] Example 17: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)thiazolidine-2-carboxamide (Compound 2'-6a)

[0360] Step 1: Preparation of (R)-2-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (Compound 2'-6a-1)

[0361] ((S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizine[1,2-b]quinoline-3,14(4H)-dione (100 mg, 0.22 mmol), (R)-3-(tert-butoxycarbonyl)thiazolidine-2-carboxylic acid (68 mg, 0.29 mmol), and DIPEA (95 mg, 0.73 mmol) were added to DMF (3 mL) and stirred for 5 minutes. HATU (139 mg, 0.37 mmol) was added, and the reaction was stirred at 25°C for 1 hour. The reaction solution was directly purified by C18 reverse phase column (acetonitrile / 0.05% formic acid aqueous solution = 10-90%). The purified solution was added with ethyl acetate (50 ml) and stirred for 5 minutes. The upper organic phase was separated and concentrated to obtain the title compound (105 mg, 0.16 mmol).

[0362] Its structural characterization data are as follows:

[0363] MS m / z(ESI):625.2[M+H] +

[0364] Step 2: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)thiazolidine-2-carboxamide (Compound 2'-6a)

[0365] (R)-2-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (105 mg, 0.16 mmol) was dissolved in 1,4-Dioxane (4 mL) and chlorinated To the mixture was added a 4M hydrogen-1,4-dioxane solution (4M, 1.3 mL), heated to 35°C for 5 hours, added methyl tert-butyl ether (5 ml), stirred, cooled to room temperature, allowed to stand, the supernatant was decanted, methyl tert-butyl ether (5 ml) was added, stirred for 15 minutes, allowed to stand, the supernatant was decanted, dried in vacuo, added acetonitrile (5 ml) and deionized water (5 ml), dissolved, and lyophilized to obtain the title compound (73 mg, 0.13 mmol, hydrochloride).

[0366] Its structural characterization data are as follows:

[0367] MS m / z(ESI):525.2[M+H] +

[0368] 1 HNMR (400MHz, DMSO) δ9.60-9.49(m,1H),8.36(d,J=8.0Hz,1H),7.93(d,J=10.8Hz,1H),7.33(s,1H),6.56(br,1H),5.49(s,2H),5.44(s,2H) ,5.26(d,J=4.0Hz,1H),4.93(d,J=5.6Hz,2H),3.51-3.44(m,2H),3.15-3.08(m,2H),2.53(s,3H),1.92-1.81(m,2H),0.87(t,J=7.2Hz,3H).

[0369] Example 18: Preparation of (S)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)methyl)thiazolidine-2-carboxamide (Compound 2'-6b)

[0370] Step 1: Preparation of (S)-2-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (Compound 2'-6b-1)

[0371] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (241.31 mg, 471.31 μmol) and (2S)-3-tert-butyloxycarbonylthiazolidine-2-carboxylic acid (100.00 mg, 428.66 μmol) were added to DMF (5 mL), followed by DIPEA (166.20 mg, 1.29 mmol) and HATU (325.95 mg, 857.32 μmol). After addition, the mixture was stirred at 22°C for 1 hour. The reaction solution was purified by HPLC and freeze-dried to obtain the title compound (145 mg, 232.12 μmol).

[0372] Its structural characterization data are as follows:

[0373] MS m / z(ESI):625.3[M+H] +

[0374] The preparation method is as follows:

[0375] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0376] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0377] Step 2: Preparation of (S)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)methyl)thiazolidine-2-carboxamide (Compound 2'-6b)

[0378] (S)-tert-Butyl 2-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)thiazolidine-3-carboxylate (145 mg, 232.12 μmol) was added to 1,4-dioxane (5 mL), followed by a 4 M hydrogen chloride-1,4-dioxane solution (2 mL). After addition, the mixture was stirred at 35°C for 2 hours. The reaction solution was purified by HPLC and freeze-dried to obtain the title compound (76.8 mg, 130.05 μmol, hydrochloride salt).

[0379] Its structural characterization data are as follows:

[0380] MS m / z(ESI):525.2[M+H] +

[0381] 1 H NMR (400MHz, DMSO) δ8.92(t,J=5.2Hz,1H),8.35(d,J=8.4Hz,1H),7.90(d,J=10.8Hz,1H),7.31(s,1H),6.53(s,1H),5.50-5.38(m,4H),4 .93-4.76(m,3H),3.28-3.00(m,3H),2.82-2.73(m,1H),2.72-2.63(m,1H),2.55-2.45(s,3H),1.94-1.78(m,2H),0.87(t,J=7.2Hz,3H).

[0382] The preparation method is as follows:

[0383] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0384] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0385] Example 19: Preparation of (S)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)thiazolidine-4-carboxamide (Compound 2'-9a)

[0386] Step 1: Preparation of (S)-4-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (Compound 2'-9a-1)

[0387] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (80 mg, 195.40 μmol), (S)-3-(tert-butyloxycarbonyl)thiazolidine-4-carboxylic acid (91.17 mg, 390.81 μmol), HATU (148.60 mg, 390.81 μmol) and DIPEA (126.27 mg, 977.02 μmol) were added to a DMF (6 mL) reaction system and stirred at 25°C for 18 hours. The reaction solution was purified by reverse phase chromatography (ACN / H2O=0-65%, 0.05% formic acid) and freeze-dried to obtain the title compound (70 mg, 112.06 μmol).

[0388] Its structural characterization data are as follows:

[0389] MS m / z(ESI):625.3[M+H] +

[0390] Step 2: Preparation of (S)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)thiazolidine-4-carboxamide (Compound 2'-9a)

[0391] Dissolve (S)-tert-butyl 4-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)thiazolidine-3-carboxylate (70 mg, 112.06 μmol) in HCl / 1,4-dioxane solution (8 mL) and stir at 25°C for 1 hour. The reaction mixture was concentrated to obtain the crude product, which was purified by preparative HPLC and freeze-dried to obtain the title compound (27 mg, 46.68 μmol).

[0392] Its structural characterization data are as follows:

[0393] MS m / z(ESI):525.2[M+H] +

[0394] 1 H NMR (400MHz, DMSO) δ8.65(d,J=8.4Hz,1H),8.34(d,J=8.0Hz,1H),7.88(d,J=10.8Hz,1H),7.29(s,1H),6.54(s,1H),5.50-5.35(m,4H),4 .92-4.72(m,2H),4.15-3.85(m,3H),3.08-2.98(m,1H),2.91-2.81(m,1H),2.55-2.47(s,3H),1.95-1.78(m,2H),0.88(t,J=7.2Hz,3H).

[0395] The preparation method is as follows:

[0396] Chromatographic column: Waters Xbridge Prep C18 OBD (5μm*19mm*150mm)

[0397] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0398] Example 20: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)thiazolidine-4-carboxamide (Compound 2'-9b)

[0399] Step 1: Preparation of (R)-4-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)thiazolidine-3-carboxylic acid tert-butyl ester (Compound 2'-9b-1)

[0400] (4R)-3-tert-Butyloxycarbonylthiazolidine-4-carboxylic acid (56.98 mg, 0.24 mmol), (S)-11-(aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizine[1,2-b]quinoline-3,14(4H)-dione (100 mg, 0.24 mmol), and N,N-diisopropylethylamine (94.70 mg, 0.73 mmol) were dissolved in DMF (2 mL), and HATU (139.31 mg, 0.37 mmol) was added. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was purified by reverse phase column chromatography to obtain the title compound 2'-9b-1 (70 mg, 0.106 mmol).

[0401] Its structural characterization data are as follows:

[0402] MS m / z(ESI):625.2[M+H] +

[0403] Step 2: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)thiazolidine-4-carboxamide (Compound 2'-9b)

[0404] Dissolve (R)-tert-butyl 4-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)thiazolidine-3-carboxylate (70 mg, 0.106 mmol) in 1,4-dioxane (4 mL). Cool the mixture to 0°C and add a 4 M solution of hydrogen chloride in 1,4-dioxane (2 mL). The reaction mixture was allowed to react at room temperature for 1 hour. After completion of the reaction, the solvent was concentrated to obtain a crude product. This crude product was reacted to obtain the title compound (29.45 mg, 51.44 μmol).

[0405] Its structural characterization data are as follows:

[0406] MS m / z(ESI):525.2[M+H] +

[0407] 1 H NMR (400MHz, DMSO) δ8.92(t,J=5.6Hz,1H),8.36(d,J=8.4Hz,1H),7.90(d,J=10.8Hz,1H),7.31(s,1H),6.54(s,1H),5.52-5.37(m,4H),4.90-4.7 7(m,2H),4.10-4.06(m,1H),3.99-3.89(m,2H),3.5-3.41(m,1H),3.04-2 ,99(m,1H),2.87-2,81(m,1H),1.92-1.79(m,2H),0.87(t,J=7.2Hz,3H).

[0408] The preparation method is as follows:

[0409] Chromatographic column: Agilent Prep C18OBD (5μm*19mm*150mm)

[0410] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0411] Example 21: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)-2,2-dimethylthiazolidine-4-carboxamide (Compound 2'-10b)

[0412] Step 1: Preparation of (9H-fluoren-9-yl)methyl (R)-4-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)-2,2-dimethylthiazolidine-3-carboxylate (Compound 2'-10b-1)

[0413] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizine[1,2-b]quinoline-3,14(4H)-dione (90 mg, 219.83 μmol), (R)-3-(((9H-fluoren-9-yl)methoxy)carbonyl)-2,2-dimethylthiazolidine-4-carboxylic acid (168.59 mg, 439.66 μmol), HATU (167.18 mg, 439.66 μmol) and DIPEA (142.05 mg, 1.10 mmol) were added to a DMF (5 mL) reaction system and stirred at 25°C for 1 hour. The reaction solution was prepared by HPLC and freeze-dried to obtain the title compound (45 mg, 58.08 μmol).

[0414] Its structural characterization data are as follows:

[0415] MS m / z(ESI):775.1[M+H] +

[0416] The preparation method is as follows:

[0417] Chromatographic column: Waters Xbridge Prep C18 OBD (5μm*19mm*150mm)

[0418] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0419] Step 2: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)-2,2-dimethylthiazolidine-4-carboxamide (Compound 2'-10b)

[0420] (9H-fluoren-9-yl)methyl (R)-4-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)-2,2-dimethylthiazolidine-3-carboxylate (45 mg, 58.08 μmol) was dissolved in DMF (4 mL). Diethylamine (21.20 mg, 290.38 μmol) was added and stirred at 25°C for 1 hour. The reaction solution was directly purified by preparative HPLC and freeze-dried to obtain the title compound (16 mg, 27.51 μmol).

[0421] Its structural characterization data are as follows:

[0422] MS m / z(ESI):553.1[M+H] +

[0423] 1 H NMR (400MHz, DMSO) δ8.99(t,J=5.6Hz,1H),8.38(d,J=8.0Hz,1H),7.90(d,J=10. 4Hz,1H),7.32(s,1H),6.54(s,1H),5.55-5.37(m,4H),4.94-4.84(m,2H),4.02-3 .91(m,1H),3.24-3.18(m,1H),3.12(d,J=11.6Hz,1H),2.97-2.90(m,1H),2.53- 2.47(s,3H),1.92-1.80(m,2H),1.44(s,3H),1.43(s,3H),0.87(t,J=7.2Hz,3H).

[0424] The preparation method is as follows:

[0425] Chromatographic column: Waters Xbridge Prep C18 OBD (5μm*19mm*150mm)

[0426] Mobile phase A: acetonitrile; mobile phase B: water

[0427] Example 22: Preparation of N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)isothiazolidine-3-carboxamide 1,1-dioxide (Compound 2'-12)

[0428] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizine[1,2-b]quinoline-3,14(4H)-dione (55 mg, 120.91 μmol), isothiazolidine-3-carboxylic acid 1,1-dioxide (19.97 mg, 120.91 μmol), HATU (68.96 mg, 181.36 μmol) and DIPEA (62.50 mg, 483.62 μmol) were added to a DMF (5 mL) reaction system and stirred at 25°C for 1 hour. The reaction solution was prepared by HPLC and freeze-dried to obtain the title compound (15 mg, 26.68 μmol).

[0429] Its structural characterization data are as follows:

[0430] MS m / z(ESI):557.0[M+H] +

[0431] 1 H NMR (400MHz, DMSO) δ8.86(t,J=5.6Hz,1H),8.41(d,J=8.4Hz,1H),7.90(d,J=10.8Hz ,1H),7.34(d,J=6.8Hz,1H),7.32(s,1H),6.54(br,1H),5.54-5.40(m,4H),4.91-4. 77(m,2H),4.03-3.93(m,1H),3.20-3.11(m,1H),3.02-2.92(m,1H),2.64-2.54(m,1 H),2.53-2.46(s,3H),2.18-2.06(m,1H),1.94-1.80(m,2H),0.87(t,J=7.2Hz,3H).

[0432] The preparation method is as follows:

[0433] Chromatographic column: Waters Xbridge Prep C18 OBD (5μm*19mm*150mm)

[0434] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0435] Example 23: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)morpholine-3-carboxamide (Compound 2'-16a)

[0436] Step 1: Preparation of (R)-3-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)morpholine-4-carboxylic acid tert-butyl ester (Compound 2'-16a-1)

[0437] (R)-4-(tert-Butoxycarbonyl)morpholine-3-carboxylic acid (44.06 mg, 190.52 μmol), (S)-11-(aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (60.00 mg, 146.55 μmol), and HATU (111.44 mg, 293.11 μmol) were added to DMF (2 mL), followed by the addition of DIPEA (94.70 mg, 732.76 μmol). After the addition was complete, the mixture was stirred at 22°C for 6 hours. The reaction solution was directly purified by HPLC and freeze-dried to obtain the title compound (57 mg, 91.55 μmol).

[0438] Its structural characterization data are as follows:

[0439] MS m / z(ESI):623.64[M+H] +

[0440] The preparation method is as follows:

[0441] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0442] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0443] Step 2: Preparation of (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)morpholine-3-carboxamide (Compound 2'-16a)

[0444] (R)-3-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)morpholine-4-carboxylic acid tert-butyl ester (57 mg, 91.55 μmol) was added to a 1,4-dioxane hydrochloride solution (5 mL). The reaction was stirred at 25°C for 6 hours. The reaction solution was directly prepared by high performance liquid chromatography and freeze-dried to obtain (R)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)morpholine-3-carboxamide (22.46 mg, 42.93 μmol).

[0445] Its structural characterization data are as follows:

[0446] MS m / z(ESI):523.53[M+H] +

[0447] 1 H NMR (400MHz, DMSO) δ8.77(t,J=6.0Hz,1H),8.40(d,J=8.4Hz,1H),7.89(d,J=10.8Hz,1H),7.31(s,1H),6.53(s,1H),5.45(d,J=9.2Hz,4 H),4.77-4.83(m,2H),3.65-3.69(m,2H),3.39-3.44(m,2H),2.63-2.71(m,2H),2.33(s,3H),1.81-1.91(m,2H),0.87(t,J=7.2Hz,3H).

[0448] The preparation method is as follows:

[0449] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0450] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0451] Example 24: Preparation of (S)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)morpholine-3-carboxamide (Compound 2'-16b)

[0452] Step 1: Preparation of (S)-3-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)morpholine-4-carboxylic acid tert-butyl ester (Compound 2'-16b-1)

[0453] To DMF (3 mL) was added (S)-4-(tert-butoxycarbonyl)morpholine-3-carboxylic acid (67.78 mg, 293.10 μmol), (S)-11-(aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (100 mg, 244.25 μmol), DIPEA (94.70 mg, 732.76 μmol), and HATU (144.22 mg, 379.34 μmol). The reaction was stirred at 25°C for 3 h. The reaction solution was purified by flash column chromatography (C18, 0.05% formic acid / acetonitrile) and lyophilized to obtain the title compound (87 mg, 132.74 μmol).

[0454] Its structural characterization data are as follows:

[0455] MS m / z(ESI):623.3[M+H] +

[0456] Step 2: Preparation of (S)-N-(((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)morpholine-3-carboxamide (Compound 2'-16b)

[0457] To a solution of 1,4-dioxane in hydrogen chloride (5 mL) was added tert-butyl-(S)-3-((((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)carbamoyl)morpholine-4-carboxylate (87 mg, 132.74 μmol). The reaction was stirred at 22°C for 3 h. The solvent was removed by rotary evaporation, and the residue was purified by HPLC and freeze-dried to give the formate salt of the title compound (36.88 mg, 65.01 μmol).

[0458] Its structural characterization data are as follows:

[0459] MS m / z(ESI):523.2[M+H] +

[0460] 1H NMR(400MHz,DMSO)δ8.76(t,J=5.9Hz,1H),8.42-8.36(m,1H),7.89(d,J=10.8Hz,1 H),7.31(s,1H),6.53(s,1H),5.49-5.40(m,4H),4.82(t,J=6.0Hz,2H),3.67(dd,J =10.8,3.5Hz,1H),3.58-3.48(m,1H),3.41(dd,J=10.8,8.0Hz,1H),3.27(dd,J=8. 1,3.5Hz,1H),2.75-2.57(m,2H),2.51(s,3H),1.86(m,2H),0.87(t,J=7.4Hz,3H).

[0461] The preparation method is as follows:

[0462] Chromatographic column: Waters XBridge Prep C18OBD (5μm*19mm*150mm)

[0463] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0464] Example 25: Preparation of (S)-3-amino-8-ethyl-8-hydroxy-2,8,11,14-tetrahydro-12H-cyclopenta[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-9,12(1H)-dione (Compound A-1)

[0465] Step 1: Preparation of (S)-8-ethyl-8-hydroxy-9,12-dioxo-1,2,8,9,12,14-hexahydro-11H-cyclopenta[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-3-yl trifluoromethanesulfonate (Compound A-1-2)

[0466] To (S)-8-ethyl-3,8-dihydroxy-2,8,11,14-tetrahydro-12H-cyclopenta[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-9,12(1H)-dione (0.154 g, 394.48 μmol) and 1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (166.95 mg, 591.72 μmol) was added DMF (6 mL), and triethylamine (119.75 mg, 1.18 mmol) was added dropwise. The mixture was stirred at room temperature for 1.5 hours. After completion of the reaction, the reaction solution was added dropwise to water with stirring, resulting in the precipitation of a solid. The mixture was extracted with ethyl acetate / methanol, and the organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane-(dichloromethane:methanol=3:1)=0-50%) and concentrated under reduced pressure to obtain the title compound (200 mg, 382.81 μmol).

[0467] Its structural characterization data are as follows:

[0468] MS m / z(ESI):523.0[M+H] +

[0469] Step 2: Preparation of tert-butyl (S)-(8-ethyl-8-hydroxy-9,12-dioxo-1,2,8,9,12,14-hexahydro-11H-cyclopenta[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-3-yl)carbamate (Compound A-1-3)

[0470] To tert-butyl carbamate (156.96 mg, 1.34 mmol) and (S)-8-ethyl-8-hydroxy-9,12-dioxo-1,2,8,9,12,14-hexahydro-11H-cyclopenta[de]pyrano[3',4':6,7]indolizine[1,2-b]quinolin-3-yl trifluoromethanesulfonate (70 mg, 133.98 μmol), cesium carbonate (87.31 mg, 267.97 μmol), Pd2(dba)3 (49.08 mg, 53.59 μmol) and X-Phos (25.55 mg, 53.59 μmol) was added 1,4-dioxane (10 mL), and the mixture was bubbling with nitrogen and microwaved at 130 ° C for 1.5 hours. After the reaction is complete, the reaction mixture is extracted with water and ethyl acetate. The organic phase is washed with brine, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain the crude product. The aqueous phase is adjusted to pH 4-5 with 2N HCl, extracted with ethyl acetate, and the organic phase is washed with brine, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The crude products are combined and purified by silica gel column chromatography (dichloromethane-methanol = 7-10%) and concentrated again under reduced pressure to obtain the title compound (182 mg, 111.54 μmol).

[0471] Its structural characterization data are as follows:

[0472] MS m / z(ESI):490.1[M+H] +

[0473] Step 3: Preparation of (S)-3-amino-8-ethyl-8-hydroxy-2,8,11,14-tetrahydro-12H-cyclopenta[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-9,12(1H)-dione (Compound A-1)

[0474] Tert-butyl (S)-(8-ethyl-8-hydroxy-9,12-dioxo-1,2,8,9,12,14-hexahydro-11H-cyclopenta[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-3-yl)carbamate (157.22 mg, 96.35 μmol) was dissolved in dichloromethane (5 mL). Hydrogen chloride-dioxane (4 M, 5 mL) and methanol (2 mL) were added and stirred at room temperature for 0.5 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The crude product was dissolved in methanol, purified by HPLC, and freeze-dried to obtain the title compound (13.97 mg, 35.52 μmol).

[0475] Its structural characterization data are as follows:

[0476] MS m / z(ESI):390.0[M+H] +

[0477] 1 H NMR (400MHz, DMSO) δ7.64(d,J=8.8Hz,1H),7.24(d,J=8.8Hz,1H),7.21(s,1H),6.44(s,1H),5.63(s,2H), 5.40(s,2H),5.11(s,2H),3.42-3.40(m,2H),3.18-3.15(m,2H),1.91-1.81(m,2H),0.88(t,J=7.2Hz,3H).

[0478] The preparation method is as follows:

[0479] Chromatographic column: Waters Sunfire Prep C18 OBD (5μm*19mm*150mm)

[0480] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0481] Example 26: Preparation of (S)-N-((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)-2-((methylamino)oxy)acetamide (Compound B-1)

[0482] Step 1: Preparation of tert-butyl hydroxymethylcarbamate (B-1-2)

[0483] Dissolve N-methylhydroxylamine hydrochloride (1 g, 11.97 mmol) in a mixture of THF (4 mL) and water (4 mL). Add potassium carbonate (827.41 mg, 5.99 mmol). Then cool to 5°C and dropwise add a solution of di-tert-butyl dicarbonate (2.87 g, 13.17 mmol) in THF (4 mL). After complete addition, stir at room temperature and allow to react overnight. After completion of the reaction, concentrate the reaction mixture under reduced pressure and extract with dichloromethane. The organic phase is washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude title compound (1.3 g, 8.83 mmol), which is used directly in the next step without purification.

[0484] Step 2: Preparation of methyl 2-(((tert-butoxycarbonyl)(methyl)amino)oxy)acetate (B-1-3)

[0485] Dissolve crude tert-butyl hydroxymethylcarbamate (1.3 g, 8.83 mmol) in THF (60 mL) and cool to 0-5°C under nitrogen. Add NaH (388.62 mg, 9.72 mmol, 60% purity) portionwise. Incubate for 30 minutes, then add methyl 2-bromoacetate (1.62 g, 10.60 mmol) dropwise. Incubate for 3 hours. After completion of the reaction, extract with water and ethyl acetate. The organic phase is washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford the crude title compound (1.3 g, 8.83 mmol), which is used directly in the next step without purification.

[0486] Its structural characterization data are as follows:

[0487] MS m / z(ESI):164.1[M-56+H] +

[0488] Step 3: Preparation of 2-(((tert-Butoxycarbonyl)(methyl)amino)oxy)acetic acid (B-1-4)

[0489] Methanol (20 mL) was added to methyl 2-(((tert-butoxycarbonyl)(methyl)amino)oxy)acetate (1.92 g, 8.76 mmol), and an aqueous sodium hydroxide solution (1 M, 10.91 mL) was added dropwise. The mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to remove methanol and then extracted with ethyl acetate. The aqueous phase was adjusted to pH 5-6 with 1N dilute hydrochloric acid and then extracted with ethyl acetate. The organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound (813 mg, 3.96 mmol).

[0490] Its structural characterization data are as follows:

[0491] MS m / z(ESI):150.7[M-56+H] +

[0492] 1 H NMR (400MHz, CDCl3) δ4.47(s,2H),3.15(s,3H),1.52(s,9H).

[0493] Step 4: Preparation of (S)-(2-(((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)amino)-2-oxoethoxy)(methyl)carbamic acid tert-butyl ester (B-1-5)

[0494] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (30 mg, 73.28 μmol) and 2-(((tert-butyloxycarbonyl)(methyl)amino)oxy)acetic acid (30.07 mg, 146.55 μmol) were dissolved in DMF (1 mL). PyBOP (76.26 mg, 146.55 μmol) and DIPEA (32 mg, 247.60 μmol) were added and stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was extracted with water and ethyl acetate. The organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude title compound (43.72 mg, 73.28 μmol), which was used directly in the next step without purification.

[0495] Its structural characterization data are as follows:

[0496] MS m / z(ESI):597.3[M+H] +

[0497] Step 5: Preparation of (S)-N-((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)-2-((methylamino)oxy)acetamide (B-1)

[0498] Crude product of tert-butyl (S)-(2-(((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)amino)-2-oxoethoxy)(methyl)carbamate (43.72 mg, 73.28 μmol) was dissolved in dichloromethane (2 mL), and a 4 M hydrogen chloride-1,4-dioxane solution (2 mL) was added dropwise. The mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the crude product was dissolved, purified by HPLC, and freeze-dried to obtain the title compound (4.81 mg, 9.11 μmol).

[0499] Its structural characterization data are as follows:

[0500] MS m / z(ESI):497.0[M+H] +

[0501] 1 H NMR (400MHz, DMSO) δ8.62(t,J=5.6Hz,1H),8.42(d,J=8.4Hz,1H),7.90(d,J=10.8Hz,1H),7.32(s,1H),6.76(q,J=6.8Hz,1H),6.52 (s,1H),5.48(s,2H),5.44(s,2H),4.87(d,J=6.0Hz,1H),2.53-2.52(m,3H),2.49(s,3H),1.92-1.80(m,2H),0.87(t,J=7.2Hz,3H).

[0502] The preparation method is as follows:

[0503] Chromatographic column: Waters Sunfire Prep C18OBD (5μm*19mm*150mm)

[0504] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0505] Example 27: Preparation of (S)-N-((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)-2-(methoxyamino)acetamide (Compound B-2) and (S)-2-(dimethylamino)-N-((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)acetamide (Compound E-1)

[0506] Step 1: Preparation of (S)-2-bromo-N-((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolo[1,2-b]quinolin-11-yl)methyl)acetamide (B-2-1)

[0507] (S)-11-(Aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H)-dione (40.00 mg, 97.70 μmol) and 2-bromoacetic acid (20.36 mg, 146.55 μmol) were mixed and dissolved in DMF (4 mL). Triethylamine (12.36 mg, 122.13 μmol) was added dropwise, and PyBOP (61.01 mg, 117.24 μmol) was added. The reaction was stirred at room temperature for 1 hour. The reaction solution was extracted with water and ethyl acetate, and the organic phase was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane-methanol = 0-30%) and concentrated again under reduced pressure to give the title compound (40 mg, 75.42 μmol).

[0508] Its structural characterization data are as follows:

[0509] MS m / z(ESI):530.2 / 532.2[M+H] +

[0510] Step 2: Preparation of (S)-N-((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)-2-(methoxyamino)acetamide (Compound B-2) and (S)-2-(dimethylamino)-N-((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl)methyl)acetamide (Compound E-1)

[0511] To (S)-2-bromo-N-((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolo[1,2-b]quinolin-11-yl)methyl)acetamide (15 mg, 28.28 μmol) and O-methylhydroxylamine hydrochloride (47.24 mg, 565.67 μmol) was added DMF (1 mL), followed by TEA (57.24 mg, 565.67 μmol). The mixture was stirred at room temperature for 1 hour, then heated to 80°C, sealed, and stirred overnight. After completion of the reaction, the reaction solution was filtered, and the filtrate was directly purified by HPLC and freeze-dried to obtain the title compounds B-2 (1.53 mg, 2.87 μmol) and E-1 (3.56 mg, 6.26 μmol).

[0512] The structural characterization data of compound B-2 are as follows:

[0513] MS m / z(ESI):497.0[M+H] +

[0514] 1 H NMR (400MHz, DMSO) δ8.74(t,J=5.6Hz,1H),8.40(d,J=8.0Hz,1H),7.91(d,J=10.4Hz,1H),7.31(s,1H),6.86(t,J=5.6Hz,1H),6. 54(s,1H),5.47(s,2H),5.44(s,2H),4.87(d,J=6.0Hz,1H),3.32(s,3H),2.49(s,3H),1.92-1.81(m,2H),0.87(t,J=7.2Hz,3H).

[0515] The structural characterization data of compound E-1 are as follows:

[0516] MS m / z(ESI):495.1[M+H] +

[0517] 1 H NMR (400MHz, DMSO) δ8.74(t,J=5.6Hz,1H),8.43(d,J=8.0Hz,1H),7.90(d,J=10.4Hz,1H),7.31(s,1H),6.50-6.40(m,1H),5.4 7(s,2H),5.44(s,2H),4.82(d,J=6.0Hz,1H),2.88(s,2H),2.49(s,3H),2.13(s,6H),1.92-1.81(m,2H),0.87(t,J=7.2Hz,3H).

[0518] The preparation method is as follows:

[0519] Chromatographic column: Waters Sunfire Prep C18 OBD (5μm*19mm*150mm)

[0520] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0521] Example 28: Preparation of (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolo[1,2-b]quinolin-1-yl)-5,5-dimethylthiazolidine-4-carboxamide (2-16a)

[0522] Step 1: Preparation of (9H-fluoren-9-yl)methyl (S)-4-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolo[1,2-b]quinolin-1-yl)carbamoyl)-5,5-dimethylthiazolidine-3-carboxylate (2-16a-1)

[0523] (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (97.03 mg, 182.55 μmol), (S)-3-(((9H-fluoren-9-yl)methoxy)carbonyl)-5,5-dimethylthiazolidine-4-carboxylic acid (70.00 mg, 182.55 μmol), HATU (138.82 mg, 365.10 μmol) and DIPEA (94.37 mg, 730.19 μmol) were added to the DMF (5 mL) reaction system and stirred at 25°C for 2 hours. The reaction was monitored by LC-MS. The reaction solution was purified by reverse phase purification (ACN / H2O=0-65%, 0.05% formic acid) and lyophilized to obtain a light yellow solid (140 mg, 174.81 μmol).

[0524] Its structural characterization data are as follows:

[0525] MS m / z(ESI):801.3[M+H] +

[0526] Step 2: Preparation of (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolo[1,2-b]quinolin-1-yl)-5,5-dimethylthiazolidine-4-carboxamide (2-16a)

[0527] (9H-fluoren-9-yl)methyl (S)-4-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolo[1,2-b]quinolin-1-yl)carbamoyl)-5,5-dimethylthiazolidine-3-carboxylate (50 mg, 62.43 μmol) was dissolved in DMF (3 mL). Diethylamine (22.83 mg, 312.15 μmol) was added and stirred at 25°C for 1.5 hours. After completion of the reaction, the reaction mixture was purified by HPLC to obtain the title compound (8 mg, 13.69 μmol).

[0528] Its structural characterization data are as follows:

[0529] MS m / z(ESI):579.2[M+H] +

[0530] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0531] 1 H NMR (400MHz, DMSO) δ8.60(d,J=8.4Hz,1H),7.82(d,J=10.8Hz,1H),7.31(s,1H),6.54(s,1H) ,5.70-5.63(m,1H),5.43(s,2H),5.33(d,J=19.2Hz,1H),5.12(d,J=18.8Hz,1H),4.34-4.26 (m,1H),4.04-3.95(m,1H),3.82-3.64(m,1H),3.29-3.19(m,2H),3.16-3.04(m,1H),2.41(s ,3H),2.22-2.05(m,2H),1.94-1.76(m,2H),1.54(s,3H),1.26(s,3H),0.87(t,J=7.2Hz,3H).

[0532] Example 29: Preparation of (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolo[1,2-b]quinolin-1-yl)-5,5-dimethylthiazolidine-4-carboxamide (2-16b)

[0533] To DMF (2 mL) was added (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolo[1,2-b]quinoline-10,13-dione (20 mg, 45.93 μmol), (R)-5,5-dimethylthiazolidine-4-carboxylic acid (7.40 mg, 45.93 μmol), DIPEA (23.74 mg, 183.72 μmol), and HATU (17.46 mg, 45.93 μmol). The reaction was stirred at 22°C for 2 h. The reaction mixture was purified by HPLC and lyophilized to obtain the title compound (11 mg, 16.73 μmol).

[0534] Its structural characterization data are as follows:

[0535] MS m / z(ESI):579.3[M+H] +

[0536] The preparation method is as follows:

[0537] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0538] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0539] 1 H NMR (400MHz, DMSO) δ8.56-8.58(d,J=8.3Hz,1H),7.81-7.83(d,J=11.0Hz,1H),7.31(s,1H),6.53(s,1H), 5.52-5.57(m,1H),5.43(s,2H),5.05-5.36(m,2H),4.24-4.28(m,1H),3.98-4.03(t,J=9.5Hz,1H),3.76(m 1H),3.27-3.30(m,1H),3.16-3.19(t,J=5.9Hz,2H),2.42(d,J=2.0Hz,3H),2.28- 2.06(m,1H),1.81-1.91(m,2H),1.43(s,3H),1.23(s,3H),0.88(t,J=7.3Hz,3H).

[0540] Biological evaluation

[0541] 1. Tumor Cell Proliferation Inhibition Test

[0542] 1. Inhibitory effect of the compound on HT29 cell proliferation

[0543] (1) Cell plating: First, culture HT29 tumor cells in the appropriate culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to an appropriate concentration for plating. The tumor cell sources are shown in Table 1.

[0544] Table 1. Tumor cell sources

[0545] Co-incubation of the compound of the present invention and tumor cells: Cell suspensions were added to 96-well plates respectively. After the cells adhered to the wall, bioactive molecules (compounds of the present invention) diluted with culture medium were added to the wells of the plates and incubated.

[0546] In vitro cell activity assay: After incubation, add Cell Counting-Lite TM2.0 reagent, shake and mix in the dark, and after a certain reaction time, the test can be carried out and the reading is read on a microplate reader (manufacturer: BMG). The reading of the culture medium well without cells is the background RLU, and the reading of the culture medium well with cells but without compound is the cell control RLU. Cell inhibition rate = (1-(sample RLU-background RLU) / (cell control RLU-background RLU)) × 100%. According to the four-parameter model fitting curve, the half-maximal inhibitory concentration (IC) of the compound is calculated. 50 ), and the test results are shown in Tables 2-1, 2-2, 2-3, 2-4 and 2-5.

[0547] (2) Data results

[0548] Table 2-1. Inhibitory activity of compounds on HT29 cell proliferation

[0549] Table 2-2. Inhibitory activity of compounds on HT29 cell proliferation

[0550] Table 2-3. Inhibitory activity of compounds on HT29 cell proliferation

[0551] Table 2-4. Inhibitory activity of compounds on HT29 cell proliferation

[0552] Table 2-5. Inhibitory activity of compounds on HT29 cell proliferation

[0553] The test results show that the compounds of the present invention in Tables 2-1, 2-2, 2-3, 2-4 and 2-5 have a strong proliferation inhibitory effect on HT29 colon cancer cells.

[0554] 2. Inhibitory effect of compounds on NCI-N87 cell proliferation

[0555] (1) Cell plating: First, culture NCI-N87 tumor cells in the appropriate culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to an appropriate concentration for plating. The tumor cell sources are shown in Table 3.

[0556] Table 3. Tumor cell origin

[0557] Co-incubation of the compound of the present invention and tumor cells: Cell suspensions were added to 96-well plates. After the cells adhered to the wall, bioactive molecules (compounds of the present invention) diluted with culture medium were added to the wells and incubated for 72 hours.

[0558] In vitro cell activity assay: After incubation, add Cell Counting-LiteTM 2.0 reagent, shake and mix in the dark, and after a certain reaction time, the test can be carried out and the reading is read on a microplate reader (manufacturer: BMG). The reading of the culture medium well without cells is the background RLU, and the reading of the culture medium well with cells but without compound is the cell control RLU. Cell inhibition rate = (1-(sample RLU-background RLU) / (cell control RLU-background RLU)) × 100%. According to the four-parameter model fitting curve, the half-maximal inhibitory concentration (IC) of the compound is calculated. 50 ), and the test results are shown in Tables 4-1, 4-2, 4-3, 4-4 and 4-5.

[0559] (2) Data results

[0560] Table 4-1. Inhibitory activity of compounds on NCI-N87 cell proliferation

[0561] Table 4-2. Inhibitory activity of compounds on NCI-N87 cell proliferation

[0562] Table 4-3. Inhibitory activity of compounds on NCI-N87 cell proliferation

[0563] Table 4-4. Inhibitory activity of compounds on NCI-N87 cell proliferation

[0564] Table 4-5. Inhibitory activity of compounds on NCI-N87 cell proliferation

[0565] The test results show that the compounds of the present invention in Tables 4-1, 4-2, 4-3, 4-4 and 4-5 have significant inhibitory effects on the proliferation of NCI-N87 human gastric cancer cells.

[0566] 3. Inhibitory effect of the compounds on HCC1954 cell proliferation

[0567] (1) Cell plating: First, culture HCC1954 tumor cells in the appropriate culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to an appropriate concentration for plating. The tumor cell sources are shown in Table 5.

[0568] Table 5. Tumor cell origin

[0569] Co-incubation of the compound of the present invention and tumor cells: After the cells adhere to the wall, the culture medium in the cells is removed, and the diluted bioactive molecules (compound of the present invention) are added to the above plate wells and incubated.

[0570] In vitro cell viability assay: After incubation, add Cell Counting-Lite™ 2.0 reagent to each well, oscillate to mix in the dark, and after a certain reaction time, perform the assay and read the results using a microplate reader (manufacturer: BMG). The background RLU is calculated using the readings from the wells containing culture medium without cells, and the vehicle RLU is calculated using the readings from the wells containing culture medium without compounds. Cell inhibition rate = (1-(sample RLU-background RLU) / (vehicle RLU-background RLU)) × 100%. The half-maximal inhibitory concentration (IC) of the compound is calculated using the four-parameter curve fitting model. 50 RLU (relative light unit): The test results are shown in Tables 6-1, 6-2, and 6-3.

[0571] (2) Data results

[0572] Table 6-1. Antiproliferation activity of compounds on HCC1954 cells

[0573] Table 6-2. Inhibitory activity of compounds on HCC1954 cell proliferation

[0574] Table 6-3. Inhibitory activity of compounds on HCC1954 cell proliferation

[0575] The test results show that the compounds of the present invention in Tables 6-1, 6-2 and 6-3 have an inhibitory effect on the proliferation of HCC1954 human breast cancer cells.

[0576] 4. Inhibitory effect of compounds on NCI-H1975 cell proliferation

[0577] (1) Cell plating: First, culture NCI-H1975 tumor cells in the appropriate culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to an appropriate concentration for plating. The tumor cell sources are shown in Table 7.

[0578] Table 7. Tumor cell sources

[0579] Co-incubation of the compound of the present invention and tumor cells: After the cells adhere to the wall, the culture medium in the cells is removed, and the diluted bioactive molecules (compound of the present invention) are added to the above plate wells and incubated.

[0580] In vitro cell viability assay: After incubation, add Cell Counting-Lite™ 2.0 reagent to each well, oscillate to mix in the dark, and after a certain reaction time, perform the assay and read the results using a microplate reader (manufacturer: BMG). The background RLU is calculated using the readings from the wells containing culture medium without cells, and the vehicle RLU is calculated using the readings from the wells containing culture medium without compounds. Cell inhibition rate = (1-(sample RLU-background RLU) / (vehicle RLU-background RLU)) × 100%. The half-maximal inhibitory concentration (IC) of the compound is calculated using the four-parameter curve fitting model. 50 RLU (relative light unit): The test results are shown in Tables 8-1, 8-2, 8-3, and 8-4.

[0581] (2) Data results

[0582] Table 8-1. Antiproliferation activity of compounds on NCI-H1975 cells

[0583] Table 8-2. Inhibitory activity of compounds on NCI-H1975 cell proliferation

[0584] Table 8-3. Inhibitory activity of compounds on NCI-H1975 cell proliferation

[0585] Table 8-4. Inhibitory activity of compounds on NCI-H1975 cell proliferation

[0586] The test results show that the compounds of the present invention in Tables 8-1, 8-2, 8-3 and 8-4 have an inhibitory effect on the proliferation of NCI-H1975 human non-small cell lung cancer cells.

[0587] 5. Inhibitory effect of compounds on NCI-H358 cell proliferation

[0588] (1) Cell plating: First, culture NCI-H358 tumor cells in the appropriate culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to an appropriate concentration for plating. The tumor cell sources are shown in Table 9.

[0589] Table 9. Tumor cell sources

[0590] Co-incubation of the compound of the present invention and tumor cells: After the cells adhere to the wall, the culture medium in the cells is removed, and the diluted bioactive molecules (compound of the present invention) are added to the above plate wells and incubated.

[0591] In vitro cell viability assay: After incubation, add Cell Counting-Lite™ 2.0 reagent to each well, oscillate to mix in the dark, and after a certain reaction time, perform the assay and read the results using a microplate reader (manufacturer: BMG). The background RLU is calculated using the readings from the wells containing culture medium without cells, and the vehicle RLU is calculated using the readings from the wells containing culture medium without compounds. Cell inhibition rate = (1-(sample RLU-background RLU) / (vehicle RLU-background RLU)) × 100%. The half-maximal inhibitory concentration (IC) of the compound is calculated using the four-parameter curve fitting model. 50 RLU (relative light unit): The test results are shown in Tables 10-1, 10-2, 10-3, and 10-4.

[0592] (2) Data results

[0593] Table 10-1. Antiproliferation activity of compounds on NCI-H358 cells

[0594] Table 10-2. Inhibitory activity of compounds on NCI-H358 cell proliferation

[0595] Table 10-3. Inhibitory activity of compounds on NCI-H358 cell proliferation

[0596] Table 10-4. Inhibitory activity of compounds on NCI-H358 cell proliferation

[0597] The test results show that the compounds of the present invention in Tables 10-1, 10-2, 10-3 and 10-4 have an inhibitory effect on the proliferation of NCI-H358 human non-small cell lung cancer cells.

[0598] 2. Liver microsome metabolic stability test

[0599] This study uses liver microsomes as an in vitro model to evaluate the metabolic stability of the present compound and its control compound in human and monkey liver microsomes.

[0600] Samples of the present invention compound and a control compound were added to human liver microsome and monkey liver microsome solutions, respectively, and mixed thoroughly by pipetting. PBS solution was then added, mixed, and pre-incubated at 37°C for 5 minutes. NADPH solution was then added to achieve a final compound concentration of 1 μM and a final human liver microsome and monkey liver microsome protein concentration of 0.5 mg / ml. After incubation for 0 and 15 minutes, a protein precipitant containing an internal standard was added. After centrifugation, the supernatant was quantitatively removed and added to the diluent, mixed thoroughly. The present invention compound and the control compound were semi-quantitatively detected by LC-MS / MS. The remaining percentage (%) of the parent drug in the incubation system was calculated as follows:

[0601] Original shape remaining rate = 100% × (A T15 / A T0 )

[0602] Note: A T15 : Peak area ratio of compound to internal standard in sample after 15 min incubation; A T0 : Peak area ratio of unreacted compound to internal standard.

[0603] The test results are as follows:

[0604] Table A: Remaining rate of prototype of the compounds of the present invention and the control compounds after incubation in human and monkey liver microsomes for 15 minutes

[0605] Although the specific embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that various modifications and substitutions may be made to those details based on all the teachings disclosed, and these changes are all within the scope of protection of the present invention. The full scope of the present invention is given by the appended claims and any equivalents thereof.

Claims

1. A compound or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, or prodrug thereof, wherein the compound has the structure shown below: in, to exist or not to exist; R1 is fluorine or chlorine, R2 is selected from methyl, chloro, hydroxy or amino; Alternatively, R1 and R2 together with the carbon atom to which they are attached form R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring; Ring A is selected from: X is selected from oxygen or sulfur; n is selected from 1 to 5; when When the methyl group exists, R1 is fluorine, R2 is methyl, R3 is hydrogen, and X is oxygen, ring A is not morpholinyl.

2. The compound of claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, and prodrug thereof, wherein the compound has the structure shown below: in, R1 is fluorine or chlorine, R2 is selected from methyl, chloro, hydroxy or amino; Alternatively, R1 and R2 together with the carbon atom to which they are attached form R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring; Ring A is selected from: Preferred X is selected from oxygen or sulfur; n is selected from 1 to 5; When R1 is fluorine, R2 is methyl, R3 is hydrogen, and X is oxygen, ring A is not morpholinyl.

3. The compound of any one of claims 1-2, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, and prodrug thereof, wherein the compound has the structure shown below: in, R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring; Ring A is selected from: Preferred X is selected from oxygen or sulfur; n is selected from 1 to 5; When R1 is fluorine, R2 is methyl, R3 is hydrogen, and X is oxygen, ring A is not morpholinyl.

4. The compound of claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, and prodrug thereof, wherein the compound has the structure shown below: in, R1 is fluorine or chlorine, R2 is selected from methyl, chloro, hydroxy or amino; Alternatively, R1 and R2 together with the carbon atom to which they are attached form R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring; Ring A is selected from: Preferred X is selected from oxygen or sulfur; n is selected from 1-5.

5. The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, and prodrug thereof, wherein the compound has the structure shown below: in, R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring; Ring A is selected from: Preferred X is selected from oxygen or sulfur; n is selected from 1-5.

6. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, and prodrug thereof, wherein the compound has the structure shown below: in, R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms are linked to the attached atom to form a ring; Ring A is selected from: Preferred X is selected from oxygen or sulfur; n is selected from 1-5.

7. The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, or prodrug thereof, wherein: R3 are each independently selected from hydrogen, C 1-6 Alkyl, halogen, hydroxyl, =O, C 1-6 Hydroxyalkyl, C 1-6 Amine alkyl, C 1-6 Alkoxy or C 1-6 Haloalkyl, or two R3 on adjacent atoms and the connecting atom are connected to a 3-6 membered carbocyclic ring or a 3-6 membered heterocyclic ring; Preferably, R3 are each independently selected from hydrogen, C 1-4 Alkyl, halogen; More preferably, each R3 is independently selected from hydrogen, methyl, ethyl, isopropyl, fluorine, chlorine, bromine or iodine.

8. The compound of any one of claims 1 to 7, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, or prodrug thereof, wherein: Ring A is selected from Preferably, ring A is selected from 9. The compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, and prodrug thereof, wherein: Selected from Preferably, Selected from 10. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, and prodrug thereof, wherein the compound has the structure shown below:

11. A compound or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled form, metabolite, or prodrug thereof, wherein the compound has the structure shown below:

12. A pharmaceutical composition comprising a compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotopically labeled substance, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers.

13. A medicine kit product comprising: a) at least one compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled, metabolite or prodrug thereof, or the pharmaceutical composition of claim 12 as a first therapeutic agent; b) optionally at least one additional therapeutic agent as a second therapeutic agent, or a pharmaceutical composition comprising an additional therapeutic agent as a second pharmaceutical composition; and c) optional packaging and / or instructions.

14. Use of the compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, and prodrug thereof, the pharmaceutical composition of claim 12, or the kit of claim 13, in the preparation of a medicament for treating a disease involving abnormal cell proliferation; Preferably, the disease is a tumor, such as an advanced solid tumor; Preferably, the tumor is selected from brain tumor, 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, female reproductive tract cancer, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, mast cell tumor, multiple myeloma, melanoma, glioma or sarcoma.

15. A method for treating a disease involving abnormal cell proliferation, comprising the following steps: administering a therapeutically effective amount of a compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, isotope-labeled, metabolite and prodrug thereof, or a pharmaceutical composition according to claim 12 or a kit of parts according to claim 13 to an individual in need thereof; Preferably, the disease is a tumor, such as an advanced solid tumor; Preferably, the tumor is selected from brain tumor, 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, female reproductive tract cancer, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, mast cell tumor, multiple myeloma, melanoma, glioma or sarcoma.

16. A compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled form, metabolite, and prodrug thereof, a pharmaceutical composition according to claim 12, or a kit according to claim 13, for use in treating a disease involving abnormal cell proliferation; Preferably, the disease is a tumor, such as an advanced solid tumor; Preferably, the tumor is selected from brain tumor, 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, female reproductive tract cancer, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, mast cell tumor, multiple myeloma, melanoma, glioma or sarcoma.

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