Camptothecin compounds, methods for preparing the same, and their applications

Abstract

To provide camptothecin compounds and their conjugates with novel structures, improved efficacy and improved safety.SOLUTION: The present invention provides a compound having the structure of the upper formula in the figure, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled product, metabolite or prodrug thereof. In the formula, R1 and R2 are each hydrogen, halogen, C1-6 alkyl, C1-6 alkoxyl, or the like, or are connected with adjacent carbon atoms to form a 5- to 6-membered oxygen-containing heterocyclic ring; R3 is hydrogen or is connected with an ortho-carbon atom of R1 to form a 6-membered carbocyclic ring; and A is selected from one of the lower two formulas in the figure.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This application claims priority to the following Chinese patent applications: Chinese Patent Application No. 202110159956.6 and filing date February 5, 2021; Chinese Patent Application No. 202110533304.4 and filing date May 17, 2021; Chinese Patent Application No. 202110718245.8 and filing date June 28, 2021; Chinese Patent Application No. 202110936768.X and filing date August 16, 2021; and Chinese Patent Application No. 202111355330.9 and filing date November 16, 2021. The disclosures of the aforementioned Chinese patent applications are incorporated herein by reference in their entirety.

[0002] This invention relates to camptothecin compounds having antitumor activity, their conjugates, methods for preparing them, and their applications in the medical field. [Background technology]

[0003] Camptothecin (CPT, Formula 1) is isolated from Camptotheca acuminata, a member of the Cornaceae family. It is a five-membered cyclic quinoline compound consisting of a quinoline ring (AB), a pyrrole ring (C), a pyridone ring (D), and an α-hydroxylactone ring (E), with the 20th position in an S configuration (see structural formula below). In the early 1970s, camptothecin was used in clinical practice due to its excellent anticancer activity, but clinical trials were later terminated due to serious side effects such as diarrhea and hemorrhagic cystitis.

[0004] [ka]

[0005] Research data show that camptothecin can form a ternary complex with cellular DNA topoisomerase I, thereby inhibiting DNA unwinding, leading to the blockage of DNA replication and cell death (Cancer Res. 1989, 49, 6365). Camptothecin and its derivatives exhibit potent antitumor activity in animal models with lung cancer, breast cancer, colorectal cancer, ovarian cancer, and others (Nature Review Cancer. 2006, 6, 789).

[0006] Currently, several camptothecin drugs are approved for market for the treatment of tumors (Med.Res.Rev.2015, 35, 753). Irinotecan is used to treat colorectal cancer; topotecan is used to treat ovarian cancer; and berotecan is used to treat ovarian cancer and small cell lung cancer. Camptothecin derivatives include exatecan, rubitecan, calenitecan, diflomotecan, lulutotecan, gimatecan, namithecan, simitecan, silatecan, quimitecan, and aeromothecan.

[0007] Camptothecin drugs or their derivatives often cause hematological toxicity induced by bone marrow suppression, such as neutropenia, leukopenia, thrombocytopenia, and anemia, as well as gastrointestinal side effects, such as nausea, vomiting, and diarrhea. Clinical studies have found that means to improve the safety and efficacy of camptothecin compounds include improving their pharmacokinetic properties, regulating their activity, reducing dosage, or forming antibody-conjugated drugs using their conjugates and antibodies. Therefore, there remains a high clinical demand and application value for developing camptothecin compounds and their conjugates with novel structures, improved efficacy, and improved safety. [Overview of the Initiative]

[0008] The present invention provides novel camptothecin compounds and their conjugates. The camptothecin compounds have good antitumor activity and are expected to be used in the treatment of tumor diseases; their conjugates have broad application prospects in ADC drugs. The first aspect of the present invention provides a compound, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled product, metabolite or prodrug thereof, and the compound has the structure shown below:

Chemical formula

Chemical formula

[0009] In some embodiments, the compound has the structure of formula (I): [ka] (In formula (I), R x is hydrogen, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 3~6 Selected from the group consisting of cycloalkyl and 3- to 6-membered heterocyclines; R y and R z It is not possible for it to be hydrogen at the same time as hydrogen and C 1~6 Alkyl, C 1~6 Haloalkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Alkylaminoalkyl, C 1~6 (Independently selected from the group consisting of alkoxyalkyls, 3-6 membered heterocyclylalkyls, and 3-6 membered heterocyclines) It has.

[0010] In some embodiments, R x is hydrogen, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 3~6 Selected from the group consisting of cycloalkyl and 3- to 6-membered heterocyclines; R y and R z It is not possible for it to be hydrogen at the same time as hydrogen and C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Alkylaminoalkyl, C1~6 A molecule is independently selected from the group consisting of alkoxyalkyls, 3-6 membered heterocyclylalkyls, and 3-6 membered heterocyclines.

[0011] In some embodiments, in formula (I), R x is hydrogen, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 3~6 Selected from the group consisting of cycloalkyl and 3- to 6-membered heterocyclines; R y and R z It is not possible for it to be hydrogen at the same time as hydrogen and C 2~6 Alkenyl and C 2~6 It is independently selected from the group consisting of alkynnyls.

[0012] In some embodiments, R x is hydrogen or C 1~6 Selected from the group consisting of alkyl groups.

[0013] In some embodiments, R x It is hydrogen.

[0014] In some embodiments, R y and R z It is not hydrogen at the same time, but hydrogen, [ka] A compound is independently selected from the group consisting of dimethylaminomethylene, morpholinomethylene, and methoxymethylene.

[0015] In some embodiments, R y and R z It is not hydrogen at the same time, but hydrogen, [ka] A compound is independently selected from the group consisting of dimethylaminomethylene and methoxymethylene.

[0016] In some embodiments, R y is hydrogen, R z teeth, [ka] Selected from the group consisting of dimethylaminomethylene, morpholinomethylene, and methoxymethylene.

[0017] In some embodiments, R y is hydrogen, R z teeth, [ka] Selected from the group consisting of dimethylaminomethylene and methoxymethylene.

[0018] In some embodiments, R x is hydrogen, R y is hydrogen, R z teeth, [ka] Selected from the group consisting of dimethylaminomethylene, morpholinomethylene, and methoxymethylene.

[0019] In some embodiments, R x is hydrogen, R y is hydrogen, R z teeth, [ka] Selected from the group consisting of dimethylaminomethylene and methoxymethylene.

[0020] In some embodiments, R x is hydrogen, R y is hydrogen, R z teeth, [ka] Selected from the group consisting of and dimethylaminomethylene.

[0021] In some embodiments, in formula (I), [ka] teeth, [ka] It has the following arrangement.

[0022] In some embodiments, in formula (I), [ka] teeth, [ka] It has the following arrangement.

[0023] In some embodiments, in formula (I), [ka] teeth, [ka] It has the following arrangement.

[0024] In some embodiments, in formula (I), [ka] teeth, [ka] It has the following arrangement.

[0025] In some embodiments, in formula (I), [ka] teeth, [ka] It has the following arrangement.

[0026] In some embodiments, in formula (I), [ka] teeth, [ka] It has the following arrangement.

[0027] In some embodiments, in formula (I), [ka] teeth, [ka] It has the following arrangement.

[0028] In some embodiments, in formula (I), [ka] teeth, [ka] It has the following arrangement.

[0029] In some embodiments, the compound has the structure of formula (II): [ka] (In formula (II), A’ is selected from one of the following:

Chemical formula

[0030] In some embodiments, the structure of formula (II) is the following formula (II)-1:

Chemical formula

[0031] In some embodiments, R x’ is selected from the group consisting of hydrogen, C 1~6 alkyl, C 1~6 haloalkyl, C 1~6 alkoxyalkyl, C 3~6 cycloalkyl, 3- to 6-membered heterocyclyl; R y’ and R z’is selected independently from the group consisting of hydrogen, C 1~6 alkyl, C 1~6 haloalkyl, C 1~6 alkoxyalkyl, C 3~6 cycloalkyl, 3- to 6-membered heterocyclyl, C 2~6 alkenyl, C 2~6 alkynyl, aryl, heteroaryl, or R y’ and R z’ are bonded to adjacent carbon atoms to form a 3- to 6-membered ring.

[0032] In some embodiments, the 3- to 6-membered ring is a 3- to 6-membered carbocyclic ring or a 3- to 6-membered heterocyclic ring.

[0033] In some embodiments, R x’ is selected from the group consisting of hydrogen and C 1~6 alkyl.

[0034] In some embodiments, R x’ is selected from the group consisting of hydrogen and methyl.

[0035] In some embodiments, R y’ and R z’ are independently selected from the group consisting of hydrogen, C 1~6 alkyl, C 1~6 balkoxyalkyl, C 1~6 alkylaminoalkyl, C 3~6 cycloalkyl, and C 2~6 alkenyl, or R y’ and R z’ are bonded to adjacent carbon atoms to form a 3- to 6-membered cycloalkyl.

[0036] In some embodiments, R y’ is selected from the group consisting of hydrogen and C 1~6 alkyl, R z’ is selected from the group consisting of hydrogen, C 1~6 alkyl, and C[[ID=C9]] 3~6 cycloalkyl, or R y’ and R z’These atoms bond with adjacent carbon atoms to form 3- to 6-membered rings.

[0037] In some embodiments, R y’ R is selected from the group consisting of hydrogen and methyl. z’ is selected from the group consisting of hydrogen, methyl and cyclopropyl, or R y’ and R z’ It bonds with adjacent carbon atoms to form a three-membered carbon-cyclic ring.

[0038] In some embodiments, R x’ R is selected from the group consisting of hydrogen and methyl. y’ R is selected from the group consisting of hydrogen and methyl. z’ is selected from the group consisting of hydrogen, methyl and cyclopropyl, or R y’ and R z’ It bonds with adjacent carbon atoms to form a three-membered carbon-cyclic ring.

[0039] In some embodiments, R x’ is hydrogen, R y’ R is selected from the group consisting of hydrogen and methyl. z’ is selected from the group consisting of hydrogen, methyl and cyclopropyl, or R y’ and R z’ It bonds with adjacent carbon atoms to form a three-membered carbon-cyclic ring.

[0040] In some embodiments, in formula (II), A' is [ka] That is the case.

[0041] In some embodiments, in formula (II)-1, [ka] teeth, [ka] has the arrangement of.

[0042] In some embodiments, in formula (II)-1,

Chemical formula

Chemical formula

[0043] In some embodiments, in formula (II)-1,

Chemical formula

Chemical formula

[0044] In some embodiments, in formula (II)-1,

Chemical formula

Chemical formula

[0045] In some embodiments, in formula (II)-1,

Chemical formula

Chemical formula

[0046] In some embodiments, in formula (II)-1,

Chem.

Chem.

[0047] In some embodiments, in formula (II)-1,

Chem.

Chem.

[0048] In some embodiments, in formula (II)-1,

Chem.

Chem.

[0049] In some embodiments, the compound has the structure of formula (III):

Chem.

Chem.

[0050] In some embodiments, the structure of the compound of formula (III) is as follows: (III)-1: [ka] As shown.

[0051] In some embodiments, R x’’ is hydrogen, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 3~6 Selected from the group consisting of cycloalkyl and 3- to 6-membered heterocyclines; R y’’ and R z’’ is hydrogen, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 3~6 Cycloalkyl, 4-6 membered heterocyclyl, C 2~6 Alkenil, C 2~6 Independently selected from the group consisting of alkynyl, aryl, and heteroaryl, or R y’’ and R z’’ These atoms bond with adjacent carbon atoms to form 3- to 6-membered rings.

[0052] In some embodiments, R x’’ is hydrogen and C 1~6 Selected from the group consisting of alkyl groups.

[0053] In some embodiments, R x’’ It is hydrogen.

[0054] In some embodiments, the 3- to 6-membered ring is either a 3- to 6-membered carbocyclic ring or a 3- to 6-membered heterocyclic ring.

[0055] In some embodiments, R y’’ and R z’’ These are hydrogen and C, respectively. 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 1~6 Alkylaminoalkyl, C 3~6 Independently selected from the group consisting of cycloalkyl and vinyl, or R y’’ and R z’’ These atoms bond with adjacent carbon atoms to form 3- to 6-membered rings.

[0056] In some embodiments, R y’’ is hydrogen, R z’’ is hydrogen, C 1~6 Alkyl, C 3~6 Selected from the group consisting of cycloalkyl and vinyl, or R y’’ and R z’’ It bonds with adjacent carbon atoms to form a 3- to 6-membered carbon-cyclic ring.

[0057] In some embodiments, R y’’ is hydrogen, R z’’ is selected from the group consisting of hydrogen, methyl, cyclopropyl and vinyl, or R y’’ and R z’’ It bonds with adjacent carbon atoms to form a three-membered carbon-cyclic ring.

[0058] In some embodiments, R x’’ is hydrogen, R y’’is hydrogen, R z’’ is selected from the group consisting of hydrogen, methyl, cyclopropyl and vinyl, or R y’’ and R z’’ It bonds with adjacent carbon atoms to form a three-membered carbon-cyclic ring.

[0059] In some embodiments, in formula (III), A'' is [ka] That is the case.

[0060] In some embodiments, in formula (III)-1, [ka] teeth, [ka] It has the following arrangement.

[0061] In some embodiments, in formula (III)-1, [ka] teeth, [ka] It has the following arrangement.

[0062] In some embodiments, in formula (III)-1, [ka] teeth, [ka] It has the following arrangement.

[0063] In some embodiments, in formula (III)-1, [ka] teeth, [ka] It has the following arrangement.

[0064] In some embodiments, in formula (III)-1, [ka] teeth, [ka] It has the following arrangement.

[0065] In some embodiments, in formula (III)-1, [ka] teeth, [ka] It has the following arrangement.

[0066] In some embodiments, in formula (III)-1, [ka] teeth, [ka] It has the following arrangement.

[0067] In some embodiments, in formula (III)-1, [ka] teeth, [ka] It has the following arrangement.

[0068] In some embodiments, the compound has the structure of formula (IV): [ka] (In formula (IV), R a and R b is hydrogen, halogen, C 1~6 Alkyl, C 1~6 Hydroxyalkyl, C 1~6 Alkoxyalkyl, C 1~6 Alkoxy, C 1~6 Independently selected from the group consisting of haloalkyl, hydroxyl, and cyano; or R a and R b It bonds with adjacent carbon atoms to form a 5-6 membered oxygen-containing heterocyclic ring; R c and R d is hydrogen, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 1~6 Alkylaminoalkyl, C 3~6 Cycloalkyl, 3-6 membered heterocyclyl, 3-6 membered heterocyclylalkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkynnyls, or R c and R d It bonds with adjacent carbon atoms to form a 3- to 6-membered carbon-cyclic or heterocyclic ring; R e is hydrogen, C 1~6 Alkyl, C 3~6 Cycloalkyl, C 1~6 Haloalkyl, C 1~6 Selected from the group consisting of alkoxyalkyls and C2-C5 heterocyclines; q = 0 or 1; If q=0, R c and R d (It cannot be hydrogen at the same time.) It has.

[0069] In some embodiments, R a and R b is hydrogen, halogen, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6 Independently selected from the group consisting of haloalkyl, hydroxyl, and cyano; or R a and R b It bonds with adjacent carbon atoms to form a 5-6 membered oxygen-containing heterocyclic ring; R c and R d is hydrogen, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 1~6 Alkylaminoalkyl, C 3~6 Cycloalkyl, 3-6 membered heterocyclyl, 3-6 membered heterocyclylalkyl, C 2~6 Alkenyl and C 2~6 Independently selected from the group consisting of alkynnyls, or R c and R d It bonds with adjacent carbon atoms to form a 3- to 6-membered carbon-cyclic or heterocyclic ring; R e is hydrogen, C 1~6 Alkyl, C 3~6 Cycloalkyl, C 1~6 Haloalkyl, C 1~6 Selected from the group consisting of alkoxyalkyls and 4-6 membered heterocyclines; q = 0 or 1; If q=0, R c and R d It cannot be hydrogen at the same time.

[0070] In some embodiments, in formula (IV), R a and R b is hydrogen, halogen, C 1~6 Alkyl, C1~6 Alkoxy, C 1~6 Independently selected from the group consisting of haloalkyl, hydroxyl, and cyano; or R a and R b It bonds with adjacent carbon atoms to form a 5-6 membered oxygen-containing heterocyclic ring; R c and R d is hydrogen, C 1~6 Alkyl, C 3~6 Cycloalkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, 4-6 membered heterocyclyl, C 2~6 Alkenil, C 2~6 Independently selected from the group consisting of alkynnyls, or R c and R d It bonds with adjacent carbon atoms to form a 3- to 6-membered carbon-cyclic or heterocyclic ring; R e is hydrogen, C 1~6 Alkyl, C 3~6 Cycloalkyl, C 1~6 Haloalkyl, C 1~6 Selected from the group consisting of alkoxyalkyls or 4-6 membered heterocyclines; q = 0 or 1; If q=0, R c and R d It cannot be hydrogen at the same time.

[0071] In some embodiments, R a and R b It consists of hydrogen, halogens and C 1~6 Independently selected from the group consisting of alkyls, or R a and R b It bonds with adjacent carbon atoms to form a 5-6 membered oxygen-containing heterocyclic ring.

[0072] In some embodiments, R a and R b is independently selected from the group consisting of hydrogen, fluorine, chlorine, and methyl, or R a and R b Together with the benzene rings bonded to them, [ka] (In the formula, Z is selected from the group consisting of -CH2-, -CD2-, -CH2CH2-, and -CF2-) It forms.

[0073] In some embodiments, R a is methyl, and R b is fluorine, or R a and R b Together with the benzene rings bonded to them, [ka] It forms.

[0074] In some embodiments, R c and R d is hydrogen, [ka] , C 1~6 Alkoxyalkyl and C 1~6 Independently selected from the group consisting of alkylaminoalkyl, or R c and R d It bonds with adjacent carbon atoms to form a 3- to 6-membered carbon-cyclic ring.

[0075] In some embodiments, R c is hydrogen, R d is hydrogen, [ka] Selected from the group consisting of methoxyethyl and cyclopropyl, or R c and R d It bonds with adjacent carbon atoms to form a 3- to 6-membered carbon-cyclic ring.

[0076] In some embodiments, R e is hydrogen and C1~6 Selected from the group consisting of alkyl groups.

[0077] In some embodiments, R e This is selected from the group consisting of hydrogen and isopropyl.

[0078] In some embodiments, R a is methyl, and R b is fluorine, or R a and R b Together with the benzene rings bonded to them, [ka] Forms R e R is selected from the group consisting of hydrogen and isopropyl, c is hydrogen, R d is hydrogen, [ka] Selected from the group consisting of methoxyethyl and cyclopropyl, or R c and R d It bonds with adjacent carbon atoms to form a three-membered carbon-cyclic ring.

[0079] In some embodiments, R a is methyl, and R b is fluorine, or R a and R b Together with the benzene rings bonded to them, [ka] Forms R e R is selected from the group consisting of hydrogen and isopropyl, c is hydrogen, R d is selected from the group consisting of hydrogen, methoxyethyl and cyclopropyl, or R c and R d It bonds with adjacent carbon atoms to form a three-membered carbon-cyclic ring.

[0080] In some embodiments, in formula (IV), [ka] teeth, [ka] It has the following arrangement.

[0081] In some embodiments, in formula (IV), [ka] teeth, [ka] It has the following arrangement.

[0082] In some embodiments, the compound has the structure of formula (V): [ka] (In equation (V), R is C 3~6 Cycloalkyl and C 1~6 Selected from the group consisting of alkoxys; A'' is, [ka] Select one of the following: R x’’’ is hydrogen, C 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 3~6 Selected from the group consisting of cycloalkyl and 3- to 6-membered heterocyclines; R y’’’ and R z’’’ is hydrogen, C 1~6 Alkyl, C 1~6Haloalkyl, C 1~6 Alkoxyalkyl, C 1~6 Alkylaminoalkyl, C 3~6 Cycloalkyl, 3-6 membered heterocyclyl, 3-6 membered heterocyclylalkyl, C 2~6 Alkenil, C 2~6 Independently selected from the group consisting of alkynyl, aryl, and heteroaryl, or R y’’’ and R z’’’ (It bonds with adjacent carbon atoms to form a 3-6 membered ring.) It has.

[0083] In some embodiments, R is selected from the group consisting of methoxy and cyclopropyl.

[0084] In some embodiments, the structure of the compound of formula (V) is as follows: (V)-1: [ka] As shown.

[0085] In some embodiments, the 3- to 6-membered ring is either a 3- to 6-membered carbocyclic ring or a 3- to 6-membered heterocyclic ring.

[0086] In some embodiments, R y’’’ and R z’’’ These are hydrogen and C, respectively. 1~6 Alkyl, C 1~6 Haloalkyl, C 1~6 Alkoxyalkyl, C 1~6 Alkylaminoalkyl, C 3~6 Independently selected from the group consisting of cycloalkyl and vinyl, or R y’’’ and R z’’’ It bonds with adjacent carbon atoms to form a 3- to 6-membered carbon-cyclic ring.

[0087] In some embodiments, R y’’’ and R z’’’ is hydrogen, or R y’’’ and R z’’’It bonds with adjacent carbon atoms to form a 3- to 6-membered carbon-cyclic ring.

[0088] In some embodiments, R x’’’ is hydrogen and C 1~6 Selected from the group consisting of alkyl groups.

[0089] In some embodiments, R x’’’ It is hydrogen.

[0090] In some embodiments, A''' is [ka] That is the case.

[0091] In some embodiments, R is selected from the group consisting of methoxy and cyclopropyl, x’’’ is hydrogen, R y’’’ and R z’’’ Both are hydrogen, or R y’’’ and R z’’’ It bonds with adjacent carbon atoms to form a three-membered carbon-cyclic ring.

[0092] In some embodiments, in formula (V)-1, [ka] teeth, [ka] It has the following arrangement.

[0093] In some embodiments, in formula (V)-1, [ka] teeth, [ka] It has the following arrangement.

[0094] In some embodiments, in formula (V)-1, [ka] teeth, [ka] It has the following arrangement.

[0095] In some embodiments, in formula (V)-1, [ka] teeth, [ka] It has the following arrangement.

[0096] In some embodiments, in formula (V)-1, [ka] teeth, [ka] It has the following arrangement.

[0097] In some embodiments, in formula (V)-1, [ka] teeth, [ka] It has the following arrangement.

[0098] In some embodiments, in formula (V)-1, [ka] teeth, [ka] It has the following arrangement.

[0099] In some embodiments, in formula (V)-1, [ka] teeth, [ka] It has the following arrangement.

[0100] In some embodiments, the present invention relates to the following compounds: [ka] TIFF0007911118000101.tif226149 TIFF0007911118000102.tif226149 TIFF0007911118000103.tif226149 TIFF0007911118000104.tif226149 To provide.

[0101] On the other hand, the present invention also relates to compounds of formula (VI), or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled products, metabolites or prodrugs thereof: MLED Equation (VI) (In the formula, M is the linker portion of the antibody or its antigen-binding fragment; L is a linker that connects linker sections M and E; E is a structural piece that connects L and D; D is a structural fragment of a cytotoxic drug. To provide.

[0102] In some embodiments, M has the following structure: [ka] It is selected from the group consisting of the following.

[0103] In some embodiments, M has the following structure: [ka] It is selected from the group consisting of the following.

[0104] In some embodiments, L is C 1~6 Alkylene, -N(R')-, Carbonyl, -O-, Val, Cit, Phe, Lys, D-Val, Leu, Gly, Ala, Asn, Val-Cit, Val-Ala, Val-Lys, Val-Lys(Ac), Phe-L ys, Phe-Lys(Ac), D-Val-Leu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, Ala-Ala-Ala, Val-Lys-Ala, Gly-Gly-Gly, Gly-Gly-Phe-Gly (Sequence ID 1) Gly-Gly-Gly-Gly-Gly (Sequence 2) , [ka] (In the formula, R' represents hydrogen, C 1~6 Alkyl or -(CH2CH2O) r - Represents an alkyl group containing -; r is an integer selected from 1 to 10; s is an integer selected from 1 to 10. It is a divalent structure composed of one or more selected from the group consisting of the following.

[0105] In some embodiments, L has the following structure: [ka] Selected from.

[0106] In some embodiments, L has the following structure: [ka] Selected from.

[0107] In some embodiments, E is a single bond, -NH-CH2- [ka] It is selected from the group consisting of the following.

[0108] In some embodiments, E is -NH-CH2-.

[0109] In some embodiments, the cytotoxic drug is selected from compounds according to any one of the items of the first aspect of the present invention.

[0110] In some embodiments, the cytotoxic drug is selected from the group consisting of compounds 1-1 to 1-15; 2-1 to 2-27; 3-1 to 3-26; 4-1 to 4-15; or 5-1 to 5-36 of the present invention.

[0111] In some embodiments, D is selected from structures formed by removing a hydrogen atom from the compound of the present invention.

[0112] In some embodiments, D is selected from structures formed by removing a hydrogen atom from compounds 1-1~1-15;2-1~2-27;3-1~3-26;4-1~4-15; or 5-1~5-36 of the present invention.

[0113] In some embodiments, D has the following structure: [ka] It is selected from the group consisting of the following.

[0114] In some embodiments, D has the following structure: [ka] It is selected from the group consisting of the following.

[0115] In some embodiments, MLED is the following compound: [ka] TIFF0007911118000114.tif218149 TIFF0007911118000115.tif221149 TIFF0007911118000116.tif199149 TIFF0007911118000117.tif90149 It is selected from the group consisting of the following.

[0116] In some embodiments, MLED is the following compound: [ka] It is selected from the group consisting of the following. [Modes for carrying out the invention]

[0117] definition Unless otherwise defined below in this Specification, all technical and scientific terms used herein have the same meaning as they would ordinarily be understood by those skilled in the art. References to the art used herein are intended to refer to the art as ordinarily understood in the art, including modifications of the art that are obvious to those skilled in the art or substitutions of equivalent art. The following terms are considered to be well understood by those skilled in the art, but are given below for the purpose of better illustrating the present invention.

[0118] The terms “comprising,” “including,” “having,” “containing,” and “involving” as used herein, and their variations thereof, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

[0119] As used herein, the asterisk (*) in the structural formula of a compound indicates that the marked carbon atom is a chiral carbon atom, and the present invention includes enantiomer pairs formed by chiral carbon atoms. If a compound contains two different chiral carbon atoms, the present invention includes four optical isomers formed by chiral carbon atoms.

[0120] When used herein, [ka] This indicates a bond that may be stereospecific ((R) or (S)) or non-stereospecific.

[0121] The term "alkyl" is defined as a saturated linear or branched aliphatic hydrocarbon group. In some embodiments, the alkyl group has 1 to 12 carbon atoms, for example, 1 to 6 carbon atoms. For example, as used herein, "C 1~6 The term "alkyl" refers to linear or branched alkyl groups having 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 are optionally substituted with one or more (e.g., 1, 2, or 3) preferred substituents.

[0122] The term "alkenyl" refers to a linear or branched hydrocarbon group containing at least one carbon-carbon double bond, for example, "C 2~6 Alkenil, "C 2~4This includes alkenyls, etc. Examples, but not limited to these, include vinyl, 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, and 1,4-hexadienyl.

[0123] The term "alkynyl" refers to a linear or branched hydrocarbon group containing at least one carbon-carbon triple bond, for example, "C 2~6 Alkinyl, C 4~6 This includes "alkynyl," etc. Examples, but not limited to these, include ethinyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 1,3-butadiinyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 1,3-pentadinyl, 1,4-pentadinyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, and 1,4-hexadinyl.

[0124] The term "cycloalkyl" refers to, but is not limited to, saturated cyclic hydrocarbon groups, and includes monocycloalkyl and bicycloalkyl (e.g., spirocycloalkyl, condensed cycloalkyl, and crosslinked cycloalkyl). 3~6 The term "cycloalkyl" refers to cycloalkyl compounds having 3 to 6 ring-forming carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl, which may optionally be substituted with one or more (e.g., 1, 2, or 3) preferred substituents, such as methyl-substituted cyclopropyl.

[0125] The terms "carbocyclic ring" or "carbocyrillic" refer to saturated or partially unsaturated hydrocarbon groups having a non-aromatic monocyclic or polycyclic structure, which are bonded to other parts of a compound through ring-forming carbon atoms. Examples, but not limited to, include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0126] The term "carbocyclic ring" refers to a saturated or unsaturated non-aromatic monocyclic or polycyclic (e.g., bicyclic) hydrocarbon ring (e.g., monocyclic rings such as cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, and cyclononane rings, or spiro rings, condensed or bridging ring systems (e.g., bicyclic rings including bicyclo[1.1.1]pentane, bicyclo[2.2.1]heptane, bicyclo[3.2.1]octane, or bicyclo[5.2.0]nonane, decahydronaphthalene, etc.)), which may optionally be substituted with one or more (e.g., one, two, or three) preferred substituents. The term "3- to 6-membered carbocyclic ring" refers to a carbocyclic ring containing three, four, five, or six ring-forming carbon atoms.

[0127] The term “heterocyclyl” or “heterocyclic ring” refers to a saturated or partially saturated monocyclic or polycyclic (e.g., bicyclic) non-aromatic ring structure in which the ring-forming atoms consist of a carbon atom and at least one (e.g., 1, 2, or 3) heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. Heterocyclyls can be bonded to the rest of the molecule through any ring atom, as long as the valence requirements are met. The heterocyclyls of the present invention are preferably 3- to 6-membered heterocyclyls. The term “3- to 6-membered heterocyclyl” as used herein refers to a heterocyclic group having 3 to 6 ring atoms and includes 3-membered, 4-membered, 5-membered, and 6-membered heterocyclyls, including nitrogen-containing heterocyclyls and oxygen-containing heterocyclyls such as 4- to 6-membered heterocyclyls, 4- to 6-membered nitrogen-containing heterocyclyls, and 4- to 6-membered oxygen-containing heterocyclyls. Generally, heterocyclic groups include, but are not limited to, azetidinyl, oxetanyl, tetrahydrofuryl, pyrrolidinyl, pyrrolidinonyl, imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, piperazinyl, and morpholinyl. The heterocyclyl in the present invention may optionally be substituted with one or more substituents described herein. The heterocyclyl in the present invention may optionally be condensed with one or more aromatic or non-aromatic rings.

[0128] The term "oxygen-containing heterocyclic ring" refers to a heterocyclic ring 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 heterocyclic ring. Specific examples, though not limited to these, include oxirane rings, tetrahydrofuran rings, furan rings, tetrahydropyran rings, and pyran rings. In this invention, the term "nitrogen-containing heterocyclic ring" refers to a heterocyclic ring as described above, in which one or more (e.g., 1, 2, or 3) ring atoms are nitrogen atoms.

[0129] The term "haloalkyl" refers to an alkyl group substituted with one or more (e.g., 1, 2, or 3) identical or different halogen atoms, where alkyl is as defined above. For example, the "C" used in this invention. 1~6The term "haloalkyl" refers to a haloalkyl having 1 to 6 carbon atoms. Common haloalkyls include, but are not limited to, -CH2F, -CHF2, -CF3, -CH2CF3, -CF2CF3, -CH2CH2CF3, and -CH2Cl. The haloalkyls of the present invention may be optionally substituted with one or more substituents described herein.

[0130] The term "aryl" refers to a group obtained by removing a hydrogen atom from the carbon atom of the aromatic nucleus of an aromatic hydrocarbon molecule, such as a 6- to 14-membered aryl group. Specific examples, though not limited to these, include phenyl, naphthyl, and anthracenyl groups.

[0131] The term "heteroaryl" refers to an aromatic ring group containing at least one ring member selected from the group consisting of N, O, and S. Specific examples, though not limited to these, include 5-6 member heteroaryls, 5-6 member nitrogen-containing heteroaryls, 5-6 member oxygen-containing heteroaryls, such as furyl, thienyl, pyrrolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, 1,2,3-triazinyl, 1,3,5-triazinyl, and 1,2,4,5-tetradinyl.

[0132] The term "alkoxy" refers to a group having the structure "alkyl-O-", where alkyl is defined above. For example, C 1~6 Alkoxy, C 1~4 Alkoxy, C 1~3 Alkoxy or C 1~2Examples include alkoxys. Common alkoxys include (but are not limited to) methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, and hexyloxy. The alkoxys in this invention are optionally substituted with one or more substituents described herein.

[0133] The term "alkoxyalkyl" refers to an alkyl group substituted with one or more (e.g., 1, 2, 3, or 4) alkoxy groups, where alkoxy and alkyl are defined above. For example, the "C" used in this invention 1~6 The term "alkoxyalkyl" refers to an alkyl group having 1 to 6 carbon atoms and substituted with one or more (e.g., 1, 2, 3, or 4) alkoxy groups. Common alkoxyalkyls include (but are not limited to) CH3O-CH2-, C2H5-O-CH2-, and C2H5-O-CH2CH2-.

[0134] The term "halo" or "halogen" group is defined as including F, Cl, Br, or I.

[0135] The term "nitrogen oxide" refers to an oxide (e.g., monooxide or dioxide) of at least one nitrogen atom in the structure of the compound of this application. A monooxide of nitrogen may exist as a single positional isomer or a mixture of positional isomers.

[0136] The term "substituted" means that one or more (e.g., 1, 2, 3, or 4) hydrogen atoms on a specified atom are selectively replaced by the indicated group, provided that the substitution does not exceed the normal valence of the specified atom under its current environment, and that the resulting compound is stable. Combinations of substituents and / or variable elements are permitted only if such combinations result in a stable compound.

[0137] If a substituent is described as "optionally substituted," the substituent may (1) be unsubstituted or (2) be substituted. If a carbon atom of a substituent is described as being optionally substituted with one or more substituents from the list of substituents, one or more hydrogen atoms on the carbon atom (up to any hydrogen atoms present) may be individually and / or simultaneously replaced by any individually selected substituents. If a nitrogen atom of a substituent is described as being optionally substituted with one or more substituents from the enumerated substituents, one or more hydrogen atoms on the nitrogen atom (up to any hydrogen atoms present) may be independently replaced by any selected substituents.

[0138] When a substituent is described as being "independently selected from" a certain group, each substituent is selected independently of the others. Therefore, each substituent may be identical or different from another substituent.

[0139] As used herein, the term “one or more” means, under reasonable conditions, one or more than one, for example, two, three, four, five, or ten.

[0140] When used herein, unless otherwise indicated, the bonding site of the substituent may be any preferred position of the substituent.

[0141] The term “stereoisomer” refers to an isomer formed as a result of at least one chiral center. In compounds having one or more (e.g., 1, 2, 3, or 4) chiral centers, racemic mixtures, single enantiomers, diastereomer mixtures, and individual diastereoisomers may be formed. Certain individual molecules may also exist as geometric isomers (cis / trans). Similarly, compounds of the present invention may exist as mixtures of two or more structurally distinct forms (generally called tautomers) in rapid equilibrium. Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, and the like. It will be understood that the scope of the present invention encompasses all such isomers or mixtures of them in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%).

[0142] In this specification, the carbon-carbon bond of the compound of the present invention is represented by a solid line ( [ka] ), solid line wedge shape ( [ka] ) or a dashed wedge shape ( [ka] ) may be used to indicate this. A solid line used to indicate a bond to a chiral carbon atom is intended to indicate that all possible stereoisomers of that carbon atom are present (e.g., specific enantiomers, racemic mixtures, etc.). A solid or dashed wedge shape used to indicate a bond to a chiral carbon atom is intended to indicate the stereoisomers as shown. If a racemic mixture is shown, the solid or dashed wedge shape is intended to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the present invention may exist in the form of 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 isomer, such as racemic mixtures and diastereoisomer pairs, and may consist of mixtures thereof.

[0143] The present invention encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be a single polymorph or a mixture of more than one polymorph in any proportion.

[0144] It is also understood that certain compounds of the present invention may exist in a free form for therapeutic use, 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, which can be used to directly or indirectly supply the compound of the present invention or its metabolites or residues after administration to a patient. Therefore, when “compound of the present invention” is referred to herein, it is also intended to encompass various derivative forms of the aforementioned compounds.

[0145] The pharmaceutically acceptable salts of the compounds of the present invention include their acid addition salts and base addition salts.

[0146] Suitable acid addition salts can be formed from acids capable of forming pharmaceutically acceptable salts, including aspartates, fumarates, glucoheptonates, glucons, glucurons, and hexafluorophosphates.

[0147] Suitable base addition salts can be formed from bases capable of forming pharmaceutically acceptable salts, including aluminum salts, algin salts, choline salts, and diethylamine salts.

[0148] For an overview 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 present invention are known to those skilled in the art.

[0149] The term "ester" refers to esters derived from each of the compounds of the formulas herein, and includes physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the present invention in the form of free acids or alcohols). The compounds of the present invention themselves may also be esters.

[0150] The compounds of the present invention may exist in the form of solvates, preferably hydrates, and the compounds of the present invention contain a polar solvent (e.g., particularly water, methanol, or ethanol) as structural elements of the crystal lattice of the compound. The amount of the polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratios.

[0151] The scope of the present invention further includes metabolites of the compounds of the present invention, i.e., substances formed in vivo when the compounds of the present invention are administered. Such substances may be produced, for example, from oxidation, reduction, hydrolysis, amidation, amidation, esterification, defatting, enzymatic degradation, etc. of the administered compound. Accordingly, the present invention includes metabolites of the compounds of the present invention, including compounds produced by contact of the compounds of the present invention with a mammal for a time sufficient to produce its metabolites.

[0152] The scope of the present invention may further include prodrugs of the compounds of the present invention. Typically, such prodrugs are functional group derivatives of the compounds, which are readily converted in vivo to desired therapeutically active compounds. In these cases, the term “administration” as used in the therapeutics of the present invention would include treating various diseases or conditions using one or more prodrugs of the compounds of the present invention, where the prodrugs are converted in vivo to the aforementioned compounds after administration to the subject. For example, “Design of Prodrug,” edited by H. Bundgaard, Elsevier, 1985, describes general methods for selecting and preparing suitable prodrug derivatives.

[0153] The scope of the present invention further includes isotopically labeled products of the compounds of the present invention, which are identical to the compounds of the present invention except that at least one atom thereof is replaced by an atom having the same atomic number but a different atomic mass or mass number from the atom that is naturally dominant in terms of atomic mass or mass number.

[0154] The present invention also encompasses compounds of the present invention that contain protecting groups. During any method for preparing the compounds of the present invention, it may be necessary and / or desirable to protect any sensitive or reactive groups on any molecules involved, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved by conventional protecting groups, for example, those described in Protective Groups in Organic Chemistry, edited by JFWMcOmie, Plenum Press, 1973; and TW Greene & PGMWuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991. These references are incorporated herein by reference. The protecting groups may be removed in appropriate subsequent steps using methods known in the art.

[0155] Pharmaceutical composition In a third aspect, the present invention provides a pharmaceutical composition comprising a compound described in the first or second aspect of the present invention, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled product, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers.

[0156] The term "pharmaceutical composition" refers to a composition that can be used as a pharmaceutical, and this includes a pharmacoactive ingredient (API) (or therapeutic agent) and, optionally, one or more pharmaceutically acceptable carriers. The term "pharmaceutically acceptable carrier" refers to an excipient that, when administered with a therapeutic agent, is suitable for contact with human and / or other animal tissues at a reasonable benefit / risk ratio within the bounds of sound medical judgment, without causing excessive toxicity, irritation, anaphylaxis, or other problems or complications.

[0157] The above-described pharmaceutical compositions can act systemically and / or topically, which can be achieved by a suitable dosage form. Dosage forms include, but are not limited to, tablets, capsules, lozenges, hard lozenges, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, aqueous suspensions, injectable solutions, elixirs, and syrups.

[0158] The above-described 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 product, metabolite, or prodrug thereof.

[0159] The present invention also provides a method for preparing the above-mentioned pharmaceutical compositions or corresponding dosage forms thereof, comprising the step of combining at least one compound of the present invention, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled product, metabolite, or prodrug thereof, with one or more pharmaceutically acceptable carriers.

[0160] kit products In a fourth embodiment, the present invention is a) As a first therapeutic agent, at least one compound described in the first or second aspect of the present invention, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled product, metabolite or prodrug thereof, or as a first pharmaceutical composition, a pharmaceutical composition described in the third aspect; b) optionally, as a second therapeutic agent, at least one additional therapeutic agent, or as a second pharmaceutical composition, a pharmaceutical composition comprising an additional therapeutic agent; c) Optionally, packaging and / or instructions for use We provide a kit product that includes the following features.

[0161] The above kit product may also comprise 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 product, metabolite, or prodrug thereof.

[0162] The present invention also provides a method for preparing the above-described kit, comprising the step of combining at least one compound of the present invention, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled product, metabolite or prodrug thereof, or the above-described pharmaceutical composition, with at least one additional therapeutic agent or a pharmaceutical composition comprising an additional therapeutic agent of any choice, and optional packaging and / or instructions for use.

[0163] medical use The compounds of the present invention may exhibit a potent effect in inhibiting abnormal cell proliferation.

[0164] Accordingly, this application provides the compounds of the present invention, or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled products, metabolites or prodrugs thereof, or the aforementioned pharmaceutical compositions, for use in treating diseases associated with abnormal cell proliferation.

[0165] Furthermore, this application also provides the use of the compounds of the present invention, or their pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled products, metabolites or prodrugs, or the aforementioned pharmaceutical compositions, in the manufacture of pharmaceuticals for treating diseases associated with abnormal cell proliferation.

[0166] In some embodiments, diseases associated with abnormal cell proliferation include, but are not limited to, tumors, such as progressive solid tumors.

[0167] This application also provides the use of the compounds of the present invention, or their pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled products, metabolites or prodrugs, or the aforementioned pharmaceutical compositions, in the preparation of preparations used to inhibit the proliferation of tumor cells. In certain embodiments, the preparations are used in vivo or in vitro. For example, the preparations may be administered to a subject to inhibit the proliferation of tumor cells in that subject; or the preparations may be applied in vitro to cells (e.g., cell lines or cells derived from the subject) to inhibit in vitro tumor cell proliferation.

[0168] The tumors of the present invention include (but are not limited to) brain tumors, lung cancer, squamous cell carcinoma, bladder cancer, stomach cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, cancer of the female reproductive system, in situ cancer, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colorectal cancer, testicular cancer, gastrointestinal stromal tumors, prostate tumors, mast cell tumors, multiple myeloma, melanoma, glioma, or sarcoma.

[0169] Treatment method In another embodiment, the present invention provides a method for treating a disease associated with abnormal cell proliferation, comprising the step of administering to an individual in need a therapeutically effective amount of the compound of the present invention, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled product thereof, metabolite or prodrug, or the aforementioned pharmaceutical composition.

[0170] The term "effective dose" refers to a dose sufficient to induce a biological or medical response in a cell, tissue, organ, or organism (e.g., an individual) and to achieve the desired prophylactic and / or therapeutic effect.

[0171] The dosage regimen may be adjusted to obtain the optimal desired response. For example, it may be administered as a single dose or as divided doses over time, or the dose may be proportionally reduced or increased according to the actual situation. It is recognized that for any particular individual, a specific dosage regimen will be adjusted according to the need and the professional judgment of the person administering or supervising the administration of the composition.

[0172] The dosage of the compound of the present invention depends on the individual's condition, the severity of the disease or condition, the rate of administration, the properties of the compound, and the judgment of the prescribing physician. Generally, the effective dose is about 0.001 to 10000 mg / kg of body weight / day of the subject. Where appropriate, the effective dose is about 0.01 to 1000 mg / kg of body weight / day of the subject. About 0.01 to 1000 mg / kg of body weight of the subject, usually about 0.1 to 500 mg / kg of body weight of the subject, can be administered daily, every two days, or every three days. Exemplary dosing regimens are once or more times per day, once or more times per week, or once or more times per month. In the case of multiple doses, the interval between single doses can generally be daily, weekly, monthly, or yearly. Alternatively, it can be administered as a sustained-release preparation, in which case the required frequency of administration will be reduced. The dosage and frequency of administration may be varied depending on the half-life of the drug in the subject and whether the use is for prevention or treatment. For preventative use, relatively low doses are administered over a long period at relatively infrequent intervals; for therapeutic use, relatively high doses may need to be administered at shorter intervals until the progression of the disease slows or stops, preferably until the individual shows partial or complete relief of disease symptoms, after which preventative use may be used.

[0173] The term “to treat” means to alleviate or eliminate a target disease or condition. When a subject receives a therapeutic dose of the compound of the present invention or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention, at least one indicator and sign of the subject shows observable and / or detectable remission and / or improvement, which indicates that the subject has been successfully “treated.” It is understood that treatment includes not only complete cure but also the achievement of several biological or medically relevant outcomes, even if not a complete cure.

[0174] The term "administer / dosage" refers to the process of applying a pharmaceutically active ingredient (e.g., the compound of the present invention) or a pharmaceutical composition containing a pharmaceutically active ingredient (e.g., the pharmaceutical composition of the present invention) to an individual or its cells, tissues, organs, biofluids, etc., resulting in the active pharmaceutically active ingredient or pharmaceutical composition coming into contact with the individual or its cells, tissues, organs, biofluids, etc. Common methods of administration include, but are not limited to, oral administration, subcutaneous administration, intramuscular administration, subperitoneal administration, ocular administration, nasal administration, sublingual administration, rectal administration, and vaginal administration.

[0175] The term “needs it” refers to a physician or other caregiver determining, based on various factors within the physician's or other caregiver's area of ​​expertise, that an individual needs or would benefit from a preventive and / or therapeutic procedure.

[0176] The term “individual” (or “subject”) refers to a human or a non-human animal. The Individuals of the present invention include individuals with disease and / or conditions (patients), as well as normal individuals. The Non-Human Animals of the present invention include all vertebrates, e.g., non-mammals, e.g., birds, amphibians, reptiles, etc., as well as mammals, e.g., non-human primates, livestock and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

[0177] Preparation method A fourth aspect of the present invention provides a method for synthesizing compounds.

[0178] The compound of formula (I) of the present invention can be synthesized by the following synthetic route.

[0179] [ka] (In the formula, R x , R y and R z(The above has the meaning; LG is a leaving group selected from the group consisting of methylsulfonyl, trifluoromethylsulfonyloxy, and halogen, preferably trifluoromethylsulfonyloxy or iodine).

[0180] Step 1: The compound of formula (I)-IM1 is obtained by a substitution reaction between the compound of formula (I)-SM1 and the compound of formula (I)-SM2.

[0181] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C.

[0182] In some embodiments, this step is carried out in a suitable organic solvent selected from the group consisting of halogenated hydrocarbons (e.g., dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane (1,2-DCE), etc.), nitriles (e.g., 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 in acetonitrile.

[0183] 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 may be selected from the group consisting of N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK), and pyridine (Py). The inorganic base may be selected from the group consisting of potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), cesium carbonate (Cs2CO3), and NaOH, preferably Na2CO3 or NaHCO3.

[0184] Step 2: The compound of formula (I) is obtained by a condensation reaction between the compound of formula (I)-IM1 and the compound of formula (I)-SM3.

[0185] In some embodiments, this step is carried out in the presence of a suitable condensation reagent selected from the group consisting of HATU, HBTU, EDCI, DCC, and HOBT, preferably HATU.

[0186] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0187] In some embodiments, this step is carried out in a suitable organic solvent selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, preferably N,N-dimethylformamide.

[0188] 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 may be selected from the group consisting of DIPEA, TEA, t-BuOK, and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably DIPEA.

[0189] The compound of formula (II)-1 of the present invention may be synthesized via the following synthetic route.

[0190] [ka] (In the formula, R x‘ , R y’ and R z‘The meaning is as described above; LG is a leaving group selected from the group consisting of methylsulfonyl, trifluoromethylsulfonyloxy and halogen, preferably trifluoromethylsulfonyloxy or iodine; PG is [ka] (A protecting group selected from the group consisting of the following.)

[0191] Step 1 The compound of formula (II)-IM1 is obtained by the substitution reaction of the compound of formula (II)-SM1.

[0192] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 100°C, preferably 50°C.

[0193] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably n-heptane.

[0194] Step 2 The compound of formula (II)-IM2 is obtained by the reduction reaction of the compound of formula (II)-IM1.

[0195] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a platinum catalyst.

[0196] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 100°C, preferably 60°C.

[0197] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably ethyl acetate.

[0198] Step 3 The compound of formula (II)-IM3 is obtained by the substitution reaction of the compound of formula (II)-IM2.

[0199] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 100°C, preferably 20°C.

[0200] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably ethyl acetate.

[0201] In some embodiments, this step is carried out under basic conditions, and the reagents that provide basic conditions include organic and inorganic bases. Examples of organic bases, but not limited to these, include triethylamine, pyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, and sodium bistrimethylsilylamide; examples of inorganic bases, but not limited to these, include potassium carbonate, sodium carbonate, sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, and potassium hydroxide, with triethylamine being preferred.

[0202] Step 4 The compound of formula (II)-IM4 is obtained by the coupling reaction of the compound of formula (II)-IM3.

[0203] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 70°C, 100°C, preferably 70°C.

[0204] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, water, and preferably a mixed solvent of tetrahydrofuran and water.

[0205] In some embodiments, this step is carried out under basic conditions, and the reagents that provide the basic conditions include organic and inorganic bases. Examples of organic bases, but not limited to these, include triethylamine, pyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, and sodium bistrimethylsilylamide; examples of inorganic bases, but not limited to these, include potassium carbonate, sodium carbonate, sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, and potassium hydroxide, preferably N,N-diisopropylethylamine.

[0206] Step 5 The compound of formula (II)-IM5 is obtained by the reduction reaction of the compound of formula (II)-IM4.

[0207] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a platinum catalyst.

[0208] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 40°C.

[0209] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably tetrahydrofuran.

[0210] Step 6 The compound of formula (II)-IM6 is obtained by the ring-closing reaction of the compound of formula (II)-IM5.

[0211] In some embodiments, this step is carried out at a suitable temperature, which is 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 5°C.

[0212] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, tert-butanol, preferably a mixed solvent of tetrahydrofuran and tert-butanol.

[0213] Step 7 The compound of formula (II)-IM7 is obtained by the substitution reaction of the compound of formula (II)-IM6.

[0214] In some embodiments, this step is carried out at a preferred temperature, which is 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 5°C.

[0215] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably trifluoroacetic acid.

[0216] In some embodiments, this step is carried out under basic conditions, and the reagents that provide the basic conditions include organic and inorganic bases. Examples of organic bases, but not limited to these, include triethylamine, pyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, and sodium bistrimethylsilylamide; examples of inorganic bases, but not limited to these, include potassium carbonate, sodium carbonate, sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, and potassium hydroxide, preferably potassium tert-butoxide.

[0217] Step 8 The compound of formula (II)-IM8 is obtained by the reduction reaction of the compound of formula (II)-IM7.

[0218] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a palladium catalyst.

[0219] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 20°C.

[0220] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably methanol.

[0221] Step 9 The compound of formula (II)-IM9 is obtained by the substitution reaction of the compound of formula (II)-IM8.

[0222] In some embodiments, this step is carried out at a preferred temperature, which is 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 20°C.

[0223] In some embodiments, this step is carried out under basic conditions, and the reagents that provide the basic conditions include organic and inorganic bases. Examples of organic bases, but not limited to these, include triethylamine, pyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, and sodium bistrimethylsilylamide; examples of inorganic bases, but not limited to these, include potassium carbonate, sodium carbonate, sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, and potassium hydroxide, preferably pyridine.

[0224] Step 10 The compound of formula (II)-IM10 is obtained by the hydrolysis reaction of the compound of formula (II)-IM9.

[0225] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 60°C.

[0226] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably methanol.

[0227] In some embodiments, the reaction is carried out under acidic conditions, and reagents that provide acidic conditions include hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with hydrochloric acid being preferred.

[0228] Step 11 The compound of formula (II)-IM11 is obtained by a ring-closing reaction of the compound of formula (II)-IM10 with (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-F]indridine-3,6,10(4H)-trione.

[0229] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 140°C.

[0230] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of toluene, methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably toluene.

[0231] In some embodiments, the reaction is carried out under acidic conditions, and reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with p-toluenesulfonic acid being preferred.

[0232] Step 12 The compound of formula (II)-IM12 is obtained by the hydrolysis reaction of the compound of formula (II)-IM11.

[0233] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 100°C.

[0234] In some embodiments, the reaction is carried out under acidic conditions. Examples of reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with hydrochloric acid being preferred.

[0235] Step 13 The compound of formula (II)-IM13 is obtained by a substitution reaction between the compound of formula (II)-IM12 and the compound of formula (II)-SM2.

[0236] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C.

[0237] In some embodiments, this step is carried out in a suitable organic solvent selected from the group consisting of halogenated hydrocarbons (e.g., dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane (1,2-DCE), etc.), nitriles (e.g., acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), tetrahydrofuran (THF), 1,4-dioxane (Diox), dimethyl sulfoxide (DMSO), and any combination thereof, preferably in acetonitrile.

[0238] 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 may be selected from the group consisting of N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK), and pyridine (Py). The inorganic base may be selected from the group consisting of potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), cesium carbonate (Cs2CO3), and NaOH, preferably Na2CO3 or NaHCO3.

[0239] Step 14 The compound of formula (II)-IM14 is obtained by the condensation reaction of the compound of formula (II)-IM13 and the compound of formula (II)-SM3.

[0240] In some embodiments, this step is carried out in the presence of a suitable condensation reagent selected from the group consisting of HATU, HBTU, EDCI, DCC, and HOBT, preferably HATU.

[0241] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0242] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably N,N-dimethylformamide.

[0243] In some embodiments, this step is carried out in the presence of a suitable base, the base comprising an organic base and an inorganic base, the organic base may be selected from the group consisting of DIPEA, TEA, t-BuOK and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3 and NaOH, preferably DIPEA.

[0244] Step 15 The compound of formula (II)-1 is obtained by the acid hydrolysis reaction of the compound of formula (II)-IM14.

[0245] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C.

[0246] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, and mixed solvents thereof, preferably a mixed solution of dichloromethane and methanol (volume ratio 2:1).

[0247] In some embodiments, the reaction is carried out under acidic conditions. Examples of reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with hydrochloric acid being preferred.

[0248] The synthesis method for formula (II)-SM3 is as follows: PG, [ka] If that is the case: [ka]

[0249] Step 1 The compound of formula (II)-SM3-3 is obtained by a substitution reaction between the compound of formula (II)-SM3-1 and the compound of formula (II)-SM3-2.

[0250] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 0 to 25°C.

[0251] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, and mixed solvents thereof, preferably tetrahydrofuran.

[0252] In some embodiments, this step is carried out in the presence of a suitable base, the base comprising an organic base and an inorganic base, the organic base may be selected from the group consisting of DIPEA, TEA, t-BuOK and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3 and NaOH, preferably K2CO3.

[0253] Step 2 The compound of formula (II)-SM3 is obtained by the hydrogenation reaction of the compound of formula (II)-SM3-3.

[0254] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C.

[0255] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, and mixed solvents thereof, preferably methanol.

[0256] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a palladium catalyst.

[0257] PG, [ka] And R y’ and R z’ However, if it's hydrogen: [ka]

[0258] Step 1 The compound of formula (II)-SM3 is obtained by the condensation reaction of the compound of formula (II)-SM3-4 and the compound of formula (II)-SM3-5.

[0259] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C.

[0260] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, and mixed solvents thereof, preferably N,N-dimethylformamide.

[0261] Alternatively, the compound of formula (II)-1 can be synthesized by the following synthetic route: [ka]

[0262] Step 1 The compound of formula (II)-IM15 is obtained by the condensation reaction of the compound of formula (II)-IM13 and the compound of formula (II)-SM4.

[0263] In some embodiments, this step is carried out in the presence of a suitable condensation reagent selected from the group consisting of HATU, HBTU, EDCI, DCC, and HOBT, preferably HATU.

[0264] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0265] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably N,N-dimethylformamide.

[0266] 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 may be selected from the group consisting of DIPEA, TEA, t-BuOK, and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably DIPEA.

[0267] Step 2 The compound of formula (II)-1 is obtained by removing the silicon protecting group from the compound of formula (II)-IM15.

[0268] Alternatively, the compound of formula (II) may be synthesized by the following synthetic route: [ka]

[0269] Step 1 The compound of formula (II) is obtained by a condensation reaction between the compound of formula (II)-IM13 and the compound of formula (II)-SM5.

[0270] In some embodiments, this step is carried out in the presence of a suitable condensation reagent selected from the group consisting of HATU, HBTU, EDCI, DCC, and HOBT, preferably HATU.

[0271] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0272] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably N,N-dimethylformamide.

[0273] 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 may be selected from the group consisting of DIPEA, TEA, t-BuOK, and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably DIPEA.

[0274] The compound of formula (III)-1 of the present invention can be synthesized by the following synthetic route.

[0275] [ka] (In the formula, R x’’ , R y’’ and R z’’ The meaning is as defined above; LG is a leaving group selected from the group consisting of methylsulfonyl, trifluoromethylsulfonyloxy and halogen, preferably trifluoromethylsulfonyl or chlorine; PG is [ka] (A protecting group selected from the group consisting of the following.)

[0276] Step 1 The compound of formula (III)-IM1 is obtained by the substitution reaction of the compound of formula (III)-SM1.

[0277] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 100°C, preferably 50°C.

[0278] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably n-heptane.

[0279] Step 2 The compound of formula (III)-IM2 is obtained by the reduction reaction of the compound of formula (III)-IM1.

[0280] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a platinum catalyst.

[0281] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 100°C, preferably 60°C.

[0282] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably ethyl acetate.

[0283] Step 3 The compound of formula (III)-IM3 is obtained by the substitution reaction of the compound of formula (III)-IM2.

[0284] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 100°C, preferably 20°C.

[0285] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably ethyl acetate.

[0286] In some embodiments, this step is carried out under basic conditions, and the reagents that provide basic conditions include organic and inorganic bases. Examples of organic bases, but not limited to these, include triethylamine, pyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, and sodium bistrimethylsilylamide; examples of inorganic bases, but not limited to these, include potassium carbonate, sodium carbonate, sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, and potassium hydroxide, with triethylamine being preferred.

[0287] Step 4 The compound of formula (III)-IM4 is obtained by the coupling reaction of the compound of formula (III)-IM3.

[0288] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 70°C, 100°C, preferably 70°C.

[0289] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, water, and preferably a mixed solvent of tetrahydrofuran and water.

[0290] In some embodiments, this step is carried out under basic conditions, and the reagents that provide the basic conditions include organic and inorganic bases. Examples of organic bases, but not limited to these, include triethylamine, pyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, and sodium bistrimethylsilylamide; examples of inorganic bases, but not limited to these, include potassium carbonate, sodium carbonate, sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, and potassium hydroxide, preferably N,N-diisopropylethylamine.

[0291] Step 5 The compound of formula (III)-IM5 is obtained by the reduction reaction of the compound of formula (III)-IM4.

[0292] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a platinum catalyst.

[0293] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 40°C.

[0294] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably tetrahydrofuran.

[0295] Step 6 The compound of formula (III)-IM6 is obtained by the ring-closing reaction of the compound of formula (III)-IM5.

[0296] In some embodiments, this step is carried out at a preferred temperature, which is 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 5°C.

[0297] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, tert-butanol, preferably a mixed solvent of tetrahydrofuran and tert-butanol.

[0298] Step 7 The compound of formula (III)-IM7 is obtained by the substitution reaction of the compound of formula (III)-IM6.

[0299] In some embodiments, this step is carried out at a preferred temperature, which is 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 5°C.

[0300] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably trifluoroacetic acid.

[0301] In some embodiments, this step is carried out under basic conditions, and the reagents that provide the basic conditions include organic and inorganic bases. Examples of organic bases, but not limited to these, include triethylamine, pyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, and sodium bistrimethylsilylamide; examples of inorganic bases, but not limited to these, include potassium carbonate, sodium carbonate, sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, and potassium hydroxide, preferably potassium tert-butoxide.

[0302] Step 8 The compound of formula (III)-IM8 is obtained by the reduction reaction of the compound of formula (III)-IM7.

[0303] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a palladium catalyst.

[0304] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 20°C.

[0305] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably methanol.

[0306] Step 9 The compound of formula (III)-IM9 is obtained by the substitution reaction of the compound of formula (III)-IM8.

[0307] In some embodiments, this step is carried out at a preferred temperature, which is 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 20°C.

[0308] In some embodiments, this step is carried out under basic conditions, and the reagents that provide the basic conditions include organic and inorganic bases. Examples of organic bases, but not limited to these, include triethylamine, pyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, and sodium bistrimethylsilylamide; examples of inorganic bases, but not limited to these, include potassium carbonate, sodium carbonate, sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, and potassium hydroxide, preferably pyridine.

[0309] Step 10 The compound of formula (III)-IM10 is obtained by the hydrolysis reaction of the compound of formula (III)-IM9.

[0310] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 60°C.

[0311] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably methanol.

[0312] In some embodiments, the reaction is carried out under acidic conditions, and reagents that provide acidic conditions include hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with hydrochloric acid being preferred.

[0313] Step 11 The compound of formula (III)-IM11 is obtained by a ring-closing reaction of the compound of formula (III)-IM10 with (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-F]indridine-3,6,10(4H)-trione.

[0314] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 140°C.

[0315] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of toluene, methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably toluene.

[0316] In some embodiments, the reaction is carried out under acidic conditions, and reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with p-toluenesulfonic acid being preferred.

[0317] Step 12 The compound of formula (III)-IM12 is obtained by the hydrolysis reaction of the compound of formula (III)-IM11.

[0318] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 100°C.

[0319] In some embodiments, the reaction is carried out under acidic conditions. Examples of reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with hydrochloric acid being preferred.

[0320] Step 13 The compound of formula (III)-IM13 is obtained by a substitution reaction between the compound of formula (III)-IM12 and the compound of formula (III)-SM2.

[0321] A substitution reaction is carried out to obtain the compound of formula (II)-IM13.

[0322] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C.

[0323] In some embodiments, this step is carried out in a suitable organic solvent selected from the group consisting of halogenated hydrocarbons (e.g., dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane (1,2-DCE), etc.), nitriles (e.g., 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 in acetonitrile.

[0324] 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 may be selected from the group consisting of N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK), and pyridine (Py). The inorganic base may be selected from the group consisting of potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), cesium carbonate (Cs2CO3), and NaOH, preferably Na2CO3 or NaHCO3.

[0325] Step 14 The compound of formula (III)-IM14 is obtained by the condensation reaction of the compound of formula (III)-IM13 and the compound of formula (III)-SM3.

[0326] In some embodiments, this step is carried out in the presence of a suitable condensation reagent selected from the group consisting of HATU, HBTU, EDCI, DCC, and HOBT, preferably HATU.

[0327] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0328] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably N,N-dimethylformamide.

[0329] 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 may be selected from the group consisting of DIPEA, TEA, t-BuOK, and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably DIPEA.

[0330] Step 15 The compound of formula (III)-1 is obtained by the acid hydrolysis reaction of the compound of formula (III)-IM14.

[0331] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C.

[0332] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, and mixed solvents thereof, preferably a mixed solution of dichloromethane and methanol (volume ratio 2:1).

[0333] In some embodiments, the reaction is carried out under acidic conditions. Examples of reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with hydrochloric acid being preferred.

[0334] The synthesis method for formula (III)-SM3 is as follows: PG, [ka] If that is the case: [ka]

[0335] Step 1: The compound of formula (III)-SM3-3 is obtained by the substitution reaction of the compound of formula (III)-SM3-1 and the compound of formula (II)-SM3-2.

[0336] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 0 to 25°C.

[0337] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, and mixed solvents thereof, preferably tetrahydrofuran.

[0338] 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 may be selected from the group consisting of DIPEA, TEA, t-BuOK, and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably K2CO3.

[0339] Step 2: The compound of formula (III)-SM3 is obtained by the hydrogenation reaction of the compound of formula (III)-SM3-3.

[0340] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C.

[0341] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, and mixed solvents thereof, preferably methanol.

[0342] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a palladium catalyst.

[0343] PG, [ka] If R y’ and R z’ It is hydrogen: [ka]

[0344] Step 1: The compound of formula (III)-SM3 is obtained by the condensation reaction of the compound of formula (III)-SM3-4 and the compound of formula (III)-SM3-5.

[0345] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C.

[0346] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, and mixed solvents thereof, preferably N,N-dimethylformamide.

[0347] Alternatively, the compound of formula (III)-1 may be synthesized by the following synthetic route: [ka]

[0348] Step 1: The compound of formula (III)-IM15 is obtained by the condensation reaction of the compound of formula (III)-IM13 and the compound of formula (III)-SM4.

[0349] In some embodiments, this step is carried out in the presence of a suitable condensation reagent selected from the group consisting of HATU, HBTU, EDCI, DCC, and HOBT, preferably HATU.

[0350] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0351] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably N,N-dimethylformamide.

[0352] 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 may be selected from the group consisting of DIPEA, TEA, t-BuOK, and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably DIPEA.

[0353] Step 2: The compound of formula (III)-1 is obtained by removing the silicon protecting group from the compound of formula (III)-IM15.

[0354] Alternatively, the compound of formula (III)-1 can be synthesized by the following synthetic route: [ka]

[0355] Step 1: The compound of formula (III)-1 is obtained by the condensation reaction of the compound of formula (III)-IM13 and the compound of formula (III)-SM5.

[0356] In some embodiments, this step is carried out in the presence of a suitable condensation reagent selected from the group consisting of HATU, HBTU, EDCI, DCC, and HOBT, preferably HATU.

[0357] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0358] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably N,N-dimethylformamide.

[0359] 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 may be selected from the group consisting of DIPEA, TEA, t-BuOK, and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably DIPEA.

[0360] The compound of formula (IV) of the present invention can be synthesized and prepared by the following synthetic route: (In the formula, R a , R b , R c , R d and R e The meaning is as stated above. If q=1, [ka]

[0361] Step 1 The compound of formula (IV)-IM1 is obtained by the nitration reaction of the compound of formula (IV)-SM1.

[0362] In some embodiments, this step is carried out at a suitable temperature, which is 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0363] Step 2 The compound of formula (IV)-IM2 is obtained by the hydrogenation reaction of the compound of formula (IV)-IM1.

[0364] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a palladium catalyst.

[0365] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0366] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably ethyl acetate.

[0367] Step 3 The compound of formula (IV)-IM3 is obtained by the acylation reaction of the compound of formula (IV)-IM2.

[0368] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 100°C, preferably 25°C.

[0369] Step 4 The compound of formula (IV)-IM4 is obtained by the reaction of the compound of formula (IV)-IM3 with DMF-DMA.

[0370] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 120°C.

[0371] Step 5 The compound of formula (IV)-IM5 is obtained by a substitution reaction between the compound of formula (IV)-IM4 and the compound of formula (IV)-SM2.

[0372] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C.

[0373] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, ethanol, N-methylpyrrolidone, and dimethyl sulfoxide, and is preferably ethanol.

[0374] Step 6 The compound of formula (IV)-IM6 is obtained by the reduction reaction of the compound of formula (IV)-IM5.

[0375] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which is preferably sodium borohydride.

[0376] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 0 to 25°C.

[0377] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, glacial acetic acid, methanol, and mixed solutions thereof, preferably glacial acetic acid.

[0378] Step 7 The compound of formula (IV)-IM7 is obtained by protecting the amino group of the compound of formula (IV)-IM6 with Fmoc.

[0379] Step 8 The compound of formula (IV)-IM8 is obtained by removing the acetyl protecting group from the amino group of the compound of formula (IV)-IM7.

[0380] Step 9 The compound of formula (IV)-IM9 is obtained by the ring-closing reaction of the compound of formula (IV)-IM8 under acidic conditions.

[0381] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 120°C.

[0382] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of toluene, xylene, N,N-dimethylformamide, N-methylpyrrolidone, and dimethyl sulfoxide, and is preferably toluene and xylene.

[0383] In some embodiments, this step is carried out under acidic conditions, and reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with p-toluenesulfonic acid being preferred.

[0384] Step 10 The compound of formula (IV)-IM10 is obtained by removing the Fmoc protecting group from the compound of formula (IV)-IM9.

[0385] Step 11 The compound of formula (IV) is obtained by the condensation reaction of the compound of formula (IV)-IM10 and the compound of formula (IV)-SM4.

[0386] In some embodiments, this step is carried out in the presence of a suitable condensation reagent selected from the group consisting of HATU, HBTU, EDCI, DCC, and HOBT, preferably HBTU.

[0387] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0388] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably N,N-dimethylformamide.

[0389] 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 may be selected from the group consisting of DIPEA, TEA, t-BuOK, and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably DIPEA.

[0390] Alternatively, if q=0, [ka]

[0391] LG is a leaving group selected from the group consisting of methylsulfonyl, trifluoromethylsulfonyloxy, and halogens, preferably trifluoromethylsulfonyloxy or iodine.

[0392] Step 1 The compound of formula (IV)-IM11 is obtained by the reduction reaction of the compound according to formula (IV)-SM5.

[0393] In some embodiments, this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, and rhodium catalysts, and is preferably a platinum catalyst.

[0394] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 100°C, preferably 25°C.

[0395] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate and tetrahydrofuran.

[0396] Step 2 The compound of formula (IV)-IM12 is obtained by the Friedel-Crafts acylation reaction of the compound of formula (IV)-IM11.

[0397] In some embodiments, this step is carried out at a suitable temperature, which is 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0398] Step 3 The compound of formula (IV)-IM13 is obtained by the ring-closing reaction of formula (IV)-IM12 under acidic conditions.

[0399] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 120°C.

[0400] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of toluene, xylene, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and preferably toluene and xylene.

[0401] In some embodiments, this step is carried out under acidic conditions, and reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, and methanesulfonic acid, with p-toluenesulfonic acid being preferred.

[0402] Step 4 The compound of formula (IV)-IM14 is obtained by the substitution reaction of formula (IV)-IM13.

[0403] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C.

[0404] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, ethanol, N,N-methylpyrrolidone, and dimethyl sulfoxide, and preferably dimethyl sulfoxide.

[0405] Step 5 The compound of formula (IV)-IM15 is obtained by the reduction reaction of the compound of formula (IV)-IM14.

[0406] In some embodiments, this step is carried out in the presence of a suitable reducing agent selected from the group consisting of palladium catalysts, platinum catalysts, rhodium catalysts, triphenylphosphine, and triethyl phosphite, preferably triethyl phosphite.

[0407] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 50°C, 60°C, 80°C, 100°C, preferably 80°C.

[0408] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, ethyl acetate, toluene, and mixed solutions thereof, preferably a mixed solution of methanol and toluene.

[0409] Step 6 The compound of formula (IV)-IM16 is obtained by a substitution reaction between the compound of formula (IV)-IM15 and the compound of formula (IV)-SM6.

[0410] In some embodiments, this step is carried out at a preferred temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C.

[0411] In some embodiments, this step is carried out in a suitable organic solvent selected from the group consisting of halogenated hydrocarbons (e.g., dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane (1,2-DCE), etc.), nitriles (e.g., 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 in acetonitrile.

[0412] 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 may be selected from the group consisting of N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK), and pyridine (Py). The inorganic base may be selected from the group consisting of potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), cesium carbonate (Cs2CO3), and NaOH, preferably Na2CO3 or NaHCO3.

[0413] Step 7 The compound of formula (IV) is obtained by the condensation reaction of the compound of formula (IV)-IM16 and the compound of formula (IV)-SM4.

[0414] In some embodiments, this step is carried out in the presence of a suitable condensation reagent, which may be selected from the group consisting of HATU, HBTU, EDCI, DCC, and HOBT, and is preferably HBTU.

[0415] In some embodiments, this step is carried out at a suitable temperature, which is 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0416] In some embodiments, this step is carried out in a suitable organic solvent, which may be selected from the group consisting of methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, n-heptane, n-hexane, and ethyl acetate, and is preferably N,N-dimethylformamide.

[0417] 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 may be selected from the group consisting of DIPEA, TEA, t-BuOK, and Py, and the inorganic base may be selected from the group consisting of K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably DIPEA.

[0418] The compound of formula (V)-1 of the present invention is a starting material [ka] It may be synthesized and prepared using a route similar to that of formula (III).

[0419] Beneficial effects of the invention The present invention provides camptothecin compounds represented by formulas (I) to (IV), as well as pharmaceutical compositions, preparation methods, and applications thereof. These compounds possess good antitumor activity, have the potential to address drug resistance, and can be used to treat diseases associated with abnormal cell proliferation, including, but not limited to, progressive solid tumors.

[0420] A specific model for carrying out the invention The present invention is further illustrated by the description of specific embodiments, but these do not limit the invention. Those skilled in the art can make various modifications or improvements in accordance with the teachings of the present invention without departing from the basic ideas and scope of the invention.

[0421] The abbreviations used in this invention have the following meanings.

[0422] [Table 1]

[0423] The structure of the compound described in the following examples was determined by nuclear magnetic resonance ( 1 Confirmed by 1H NMR or mass spectrometry (MS).

[0424] Nuclear magnetic resonance method ( 1 The instrument used for ¹H NMR was a Bruker 400 MHz nuclear magnetic resonance spectrometer; the internal standard was hexaduteriodimethyl sulfoxide (DMSO-d6); and the internal standard was tetramethylsilane (TMS).

[0425] The abbreviations used in the nuclear magnetic resonance (NMR) spectra of the examples are shown below.

[0426] s: singlet, d: doublet, t: triplet, q: quadruplet, m: multiplet, br: broadband, J: coupling constant, Hz: Hertz, DMSO-d6: deuterated dimethyl sulfoxide. The δ value is expressed in ppm.

[0427] The mass spectrometer (MS) used was an Agilent (ESI) mass spectrometer, the Agilent 6120B.

[0428] Example 1: (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]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypenta-3-inamide and (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]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypenta-3-inamide [ka]

[0429] Compound 2-hydroxypenta-3-ic acid (4.29 mg, 37.63 μmol) was dissolved in DMF (1 mL), and HATU (21.46 mg, 56.44 μmol), SM1-1 (10.00 mg, 22.94 μmol), and DIPEA (7.29 mg, 56.44 μmol) were added. The mixture was reacted at 25°C for 2 hours. The reaction solution was concentrated under reduced pressure, and the concentrate was directly purified by preparative high-performance liquid chromatography (under the conditions described below) to obtain the title compounds 1-1-A (3.24 mg) and 1-1-B (3.98 mg).

[0430] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 2]

[0431] Retention time: 1-1-A: 10.8 minutes; 1-1-B: 11.1 minutes.

[0432] The structural characteristics data for 1-1-A were as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.58 (d, J = 8.4Hz, 1H), 7.78 (d, J = 11.2 Hz, 1H), 7.31 (s, 1H), 6.54 (s, 1H), 6.17 (d, J = 6.0Hz, 1H), 5.58-5.47 (m, 1H), 5.42 (s, 2H), 5.23 (s, 2H), 4.73-4.64 (m, 1H), 3.25-3.06 (m, 2H), 2.39 (s, 3H), 2.27-2.05 (m, 2H), 1.96-1.77 (m, 5H), 0.87 (t,J = 7.2Hz, 3H). ESI-MS (m / z): 532.2 [M+H] + . 1-1-B's structural characteristics are as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.62 (d, J = 8.8Hz, 1H), 7.78 (d, J = 10.8 Hz, 1H), 7.31 (s, 1H), 6.54 (s, 1H), 6.19 (d, J =6.0 Hz, 1H), 5.58-5.47 (m, 1H), 5.42 (s, 2H), 5.21 (d, J = 4.8 Hz, 2H), 4.73-4.65 (m, 1H), 3.27- 3.06 (m, 2H), 2.39 (s, 3H), 2.27-2.05 (m, 2H),1.93-1.80 (m, 2H), 1.80 (d, J = 2.0 Hz, 3H), 0.87 (t, J = 7.2Hz, 3H). ESI-MS (m / z): 532.2 [M+H] + .

[0433] Example 2: (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]indolidino[1,2-b]quinoline-1-yl)-2-hydroxy-3-enamide and (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]indolidino[1,2-b]quinoline-1-yl)-2-hydroxy-3-enamide [ka]

[0434] Ethylene glycolic acid (9.61 mg, 94.07 μmol) was dissolved in DMF (2 mL), and HATU (44.70 mg, 117.58 μmol), compound SM1-1 (25.00 mg, 0.047 mmol), and DIPEA (24.30 mg, 188.13 μmol) were added. The mixture was reacted at 25°C for 2 hours. The reaction solution was concentrated under reduced pressure, and the concentrate was directly purified by preparative high-performance liquid chromatography to obtain the title compounds 1-7-A (4.00 mg) and 1-7-B (1.38 mg).

[0435] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 3]

[0436] Retention time: 1-7-A: 8.7 minutes; 1-7-B: 9.1 minutes.

[0437] The structural characteristics data for 1-7-A were as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.51 (s, J = 8.8Hz, 1H), 7.76 (s, J = 10.8 Hz, 1H), 7.29 (s, 1H), 6.52 (s, 1H), 6.15 -6.04 (m,1H), 5.58-5.49 (m, 1H), 5.42 (s, 2H), 5.39 (s, 1H), 5.24-5.01 (m, 3H), 4.54 (s,J = 4.9 Hz, 1H), 3.24-3.05 (m, 2H), 2.37 (s, 3H), 2.16 (s, 2H), 1.91-1.79 (m,2H), 0.87 (t, J = 7.2 Hz, 3H). ESI-MS (m / z): 520.1 [M+H] + . 1-7-B's structural characteristics are as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.48 (s, 1H), 7.78(s, 1H), 7.30 (s, 1H), 6.12-5.92 (m, 1H), 5.54-5.47 (m, 1H), 5.42 (s, 2H), 5.37(dt, J = 1.7 Hz, 1H), 5.18 (s, 2H), 5.16-5.14 (m, 1H), 4.5-4.52 (m, 1H), 3.22-3.08 (m, 2H), 2.38 (s, 3H), 2.25-2.16 (m, 1H), 2.16-2.06 (m, 1H),1.93-1.79 (m, 2H), 0.87 (t, J = 7.2 Hz, 3H). ESI-MS (m / z): 520.1 [M+H] + .

[0438] Example 3: N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide and N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide [ka]

[0439] Step 1: Synthesis of 1-chloro-3-bromo-2-methyl-5-nitrobenzene Compound 2-1-01 (5.00 g, 29.14 mmol) was dissolved in n-heptane (25 mL) at 25 °C, concentrated sulfuric acid (25 mL) was added, and the mixture was heated to 50 °C. In a batch, NBS (6.22 g, 34.97 mmol) was added at 50 °C and the mixture was reacted at 50 °C for 2 hours. The reaction was monitored by thin-layer chromatography (ethyl acetate:petroleum ether = 1:10), the reaction solution was cooled to room temperature, added dropwise to ice water, extracted with toluene, the organic phases were combined, washed with sodium sulfite solution, water and saturated brine, dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure, the crude product was purified by preparative high-performance liquid chromatography, and the fraction was freeze-dried to obtain 4.88 g of the title compound.

[0440] Chromatography column: C18 ODS 45mm x 450mm x 8.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 4]

[0441] Step 2: Synthesis of 3-chloro-5-bromo-4-methylaniline At 25°C, compound 2-1-02 (4.88 g, 19.48 mmol) was dissolved in ethyl acetate (100 mL), carbon-supported platinum (2.00 g, 19.48 mmol, 5% content) was added, and the mixture was subjected to hydrogen displacement. The reaction was carried out at 60°C for 4 hours under the protection of a hydrogen balloon, and the reaction was monitored by LC-MS. The reaction solution was filtered, and the filtrate was concentrated to obtain 3.68 g of the crude title compound, which was used directly in the next reaction without further purification.

[0442] Step 3: Synthesis of N-(3-chloro-5-bromo-4-methylphenyl)acetamide At 20°C, compound 2-1-03 (3.63 g, 14.82 mmol) was dissolved in ethyl acetate (70 mL), triethylamine (4.50 g, 44.45 mmol) and acetic anhydride (2.27 g, 22.23 mmol) were added, and the reaction was allowed to proceed at 20°C for 20 hours, with the reaction monitored by LC-MS. Water was added to the reaction solution, extracted with ethyl acetate, the organic phases were combined, dehydrated with anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. This was then slurryed in a mixed solvent of ethyl acetate:petroleum ether = 1:5 to obtain 2.86 g of the title compound.

[0443] Step 4: Synthesis of (Z)-4-(5-acetamido-3-chloro-2-methylphenyl)buta-3-enoic acid At 20°C, compound 2-1-04 (1.80 g, 6.86 mmol) was dissolved in THF (20 mL) and water (5 mL). Vinylacetic acid (708.31 mg, 8.23 ​​mmol), DIPEA (1.95 g, 15.08 mmol), and tris(o-methylphenyl)phosphorus (62.60 mg, 0.20 mmol) were added, the reaction system was subjected to nitrogen purging, and then heated to 70°C and reacted for 5 hours. The reaction was monitored by LC-MS. 1N sodium hydroxide solution was added to the reaction solution to adjust the pH to 8, and ethyl acetate was added for extraction. The aqueous phase was adjusted to pH=3 with 1N hydrochloric acid, extracted with ethyl acetate, and combined with the organic phase. The mixture was dehydrated with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 0.82 g of the title compound, which was used directly in the next reaction.

[0444] Step 5: Synthesis of 4-(5-acetamido-3-chloro-2-methylphenyl)butanoic acid At 20°C, compound 2-1-05 (2.60 g, 9.71 mmol) was dissolved in THF (50 mL), Pd / C (0.52 g, 10% content) was added, the system was subjected to hydrogen displacement, and the reaction was carried out at 40°C for 2 hours under the protection of a hydrogen balloon, with the reaction monitored by LC-MS. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain 2.43 g of the title compound, which was used directly in the next reaction without further purification.

[0445] Step 6: Synthesis of N-(3-chloro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide Compound 2-1-06 (2.43 g, 9.01 mmol) was dissolved in trifluoroacetic acid (10 mL), cooled to 5°C, and anhydrous trifluoroacetic acid (3.78 g, 18.02 mmol, 2.50 mL) was added dropwise. The reaction was allowed to proceed at 5°C for 4 hours, and the reaction was monitored by LC-MS. The reaction solution was added to water, the pH was adjusted to 9 with 10N sodium hydroxide, extracted with ethyl acetate, the organic phases were combined, dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash silica gel column (ethyl acetate:petroleum ether = 0-20%) to obtain 1.53 g of the title compound.

[0446] Step 7: Synthesis of (Z)-N-(3-chloro-7-(hydroxyimino)-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide At 5°C, potassium tert-butoxide (1.50 g, 13.37 mmol) was dissolved in THF (16 mL) and tert-butanol (4 mL). Compound 2-1-07 (1.53 g, 6.08 mmol) from the THF solution (16 mL) was added dropwise, followed by the addition of amyl nitrite (1.14 g, 9.73 mmol) dropwise after 10 minutes. The reaction was allowed to proceed at 5°C for 1 hour, and the reaction was monitored by LC-MS. The reaction solution was adjusted to pH=5 with 1N hydrochloric acid, extracted with ethyl acetate, and the combined organic phase was dehydrated with anhydrous sodium sulfate. The mixture was concentrated under reduced pressure, and the concentrate was slurryed with methyl tert-butyl ether to obtain 1.20 g of the title compound.

[0447] Step 8: N-(7-amino-3-chloro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide At 20°C, compound 2-1-08 (0.50 g, 1.78 mmol) was dissolved in methanol (8 mL) and 2N hydrochloric acid (8 mL). Pd / C (0.15 g, 10% content) was added, the system was subjected to hydrogen displacement, and the reaction was carried out at 5°C for 2 hours under the protection of a hydrogen balloon. The reaction was monitored by LC-MS. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain 0.52 g of the hydrochloride salt of the title compound, which was used directly in the next reaction without further purification.

[0448] Step 9: Synthesis of N,N'-(3-chloro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1,7-diyl)diacetamide At 20°C, compound 2-1-09 (0.52 g, 1.70 mmol) was dissolved in pyridine (5 mL), acetic anhydride (2 mL) was added, and the reaction was allowed to proceed at 20°C for 2 hours, with the reaction monitored by LC-MS. The reaction solution was added to water, extracted with ethyl acetate, the organic phase was washed with water, and the two phases were combined. The mixture was dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was purified by flash silica gel column chromatography (ethyl acetate:petroleum ether = 0-30%) to obtain 0.22 g of the title compound.

[0449] Step 10: Synthesis of N-(8-amino-6-chloro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)acetamide At 20°C, compound 2-1-10 (0.45 g, 1.46 mmol) was dissolved in methanol (16 mL), 2N hydrochloric acid (16 mL) was added, and the mixture was heated to 60°C and reacted for 2 hours. The reaction was monitored by LC-MS. The reaction solution was cooled, saturated sodium bicarbonate solution was added to adjust the pH to 8, and the mixture was extracted with ethyl acetate. The organic phases were combined, dehydrated with anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 0.23 g of the title compound, which was used directly in the next step without further purification.

[0450] Step 11: Synthesis of N-((9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzopyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)acetamide Compound 2-1-11 (0.23 g, 0.78 mmol) was dissolved in toluene (10 mL), and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolidine-3,6,10(4H)-trione (0.23 g, 0.87 mmol) and p-toluenesulfonic acid (26.73 mg, 0.16 mmol) were added. The mixture was heated to 140 °C and reacted for 5 hours, and the reaction was monitored by LC-MS. The reaction solution was concentrated, and the crude product was purified by flash silica gel column (methanol:dichloromethane = 0-10%) to obtain 0.15 g of the title compound.

[0451] Step 12: Synthesis of (9S)-1-amino-5-chloro-9-ethyl-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzenepyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione Compound 2-1-12 (40.00 mg, 0.08 mmol) was added to concentrated hydrochloric acid (1 mL), heated to 100°C, and reacted for 5 hours. The reaction was monitored by LC-MS. The reaction solution was filtered, the filtrate was purified by preparative high-performance liquid chromatography, and the fraction was freeze-dried to obtain 12.00 mg of trifluoroacetate of the title compound 2-23.

[0452] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% trifluoroacetic acid) [Table 5]

[0453] The structural characteristics data was as follows: ESI-MS (m / z): 452.1[M+H] + .

[0454] Step 13: Synthesis of 2-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)acetamide and 2-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)acetamide At 25°C, the trifluoroacetate salt of compound 2-23 (40.00 mg, 81.91 μmol) was dissolved in N,N-dimethylformamide (1 mL). Then, 2-((tert-butyldiphenylsilyl)oxy)acetic acid (30.91 mg, 98.29 μmol), HATU (62.25 mg, 163.81 μmol), and N,N-diisopropylethylamine (42.34 mg, 327.63 μmol) were added sequentially, and the reaction was allowed to proceed at 25°C for 0.5 hours. The reaction was monitored by LC-MS. Once the reaction was complete, water was added to the reaction solution, and it was extracted with dichloromethane / methanol (volume / volume = 10 / 1). The organic phases were combined, dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified and separated by preparative thin-layer chromatography (dichloromethane:methanol = 20:1) to obtain two isomers. The two isomers were named 2-1-13-A (15.00 mg, Rf value 0.3) and 2-1-13-B (12.00 mg, Rf value 0.35) according to their Rf values.

[0455] Step 14: Synthesis of N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide and N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide At 25°C, 2-1-13-A (15.00 mg) and 2-1-13-B (12.00 mg) were dissolved in tetrahydrofuran (1 mL) in two separate reaction flasks. A mixed solution of tetrabutylammonium fluoride (1 M in tetrahydrofuran) / glacial acetic acid (vol / vol = 13 / 1) (50 μL) was added dropwise, and the reaction was allowed to proceed at 25°C for 0.5 hours, with the reaction monitored by LC-MS. Upon completion of the reaction, the reaction solution was purified by preparative high-performance liquid chromatography, and the fractions were freeze-dried to obtain the title compounds 2-1-A (6.94 mg) and 2-1-B (4.00 mg).

[0456] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 6]

[0457] The structural characteristics data for 2-1-A were as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.43 (d, J = 8.8Hz, 1H), 8.16 (s, 1H), 7.31 (s, 1H), 6.55 (s, 1H), 5.65-5.36 (m, 4H), 5.21 (q,J = 19.0 Hz, 2H), 3.95 (d, J = 5.7 Hz, 2H), 3.26-3.11 (m, 2H), 2.53 (s, 3H), 2.30-2.08 (m, 2H), 1.94-1.79 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 510.1 [M+H] + . 2-1-B's structural characteristics are as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.45 (d, J = 8.9Hz, 1H), 8.15 (s, 1H), 7.31 (s, 1H), 6.54 (s, 1H), 5.64-5.35 (m, 4H), 5.19 (q,J = 19.0 Hz, 2H), 3.97 (d, J = 5.2 Hz, 2H), 3.27-3.10 (m, 2H), 2.51 (s, 3H), 2.27-2.10 (m, 2H), 1.93-1.80 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 510.1 [M+H] + .

[0458] Example 4: (2S)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine and (2S)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine [ka]

[0459] Step 1: (2S)-2-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)propylamide Synthesis of (2S)-2-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)propylamine At 25°C, 30.00 mg (61.43 μmol) of the hydrochloride salt of 2-23 was dissolved in 1 mL of N,N-dimethylformamide. (S)-2-((tert-butyldiphenylsilyl)oxy)propionic acid (24.21 mg, 73.72 μmol), HATU (35.01 mg, 92.14 μmol), and N,N-diisopropylethylamine (23.82 mg, 184.29 μmol) were added sequentially, and the reaction was allowed to proceed at 25°C for 1 hour. The reaction was monitored by LC-MS. Once the reaction was complete, water was added to the reaction solution, and it was extracted with dichloromethane / methanol (volume / volume = 10 / 1). The organic phases were combined, dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified and separated by preparative thin-layer chromatography (dichloromethane:methanol = 15:1) to obtain two isomers. The two isomers were named 2-7-01-A (6.00 mg, Rf value 0.35) and 2-7-01-B (6.00 mg, Rf value 0.40) according to their Rf values.

[0460] Step 2: Synthesis of (2S)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine and (2S)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine At 25°C, 2-7-01-A (6.00 mg, 7.87 μmol) and 2-7-01-B (6.00 mg, 7.87 μmol) were dissolved in anhydrous THF (1 mL) in two separate reaction flasks. A mixed solution of tetrabutylammonium fluoride (1 M in tetrahydrofuran) / glacial acetic acid (volt / volt = 13 / 1) (50 μL) was added dropwise, and the reaction was allowed to proceed at 25°C for 0.5 hours, with the reaction monitored by LC-MS. Upon completion of the reaction, the reaction solution was purified by preparative high-performance liquid chromatography, and the fractions were freeze-dried to obtain the title compounds 2-7-A (2.50 mg) and 2-7-B (3.00 mg).

[0461] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 7]

[0462] The structural characteristics data for 2-7-A were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.38 (d, J = 8.8Hz, 1H), 8.15 (s, 1H), 7.31 (s, 1H), 6.55 (s, 1H), 5.56 - 5.47 (m, 2H), 5.42(s, 2H), 5.26 - 5.11 (m, 2H), 4.17 - 4.06 (m, 1H), 3.27 - 3.10 (m, 2H), 2.52(s, 3H), 2.27 - 2.08 (m, 2H), 1.86 (tt, J = 14.1, 7.3 Hz, 2H), 1.30 (d, J = 6.7Hz, 3H), 0.87 (t, J = 7.3 Hz (3H). ESI-MS (m / z): 524.2 [M+H] + . The structural characteristics data for 2-7-B were as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 9.1Hz, 1H), 8.12 (s, 1H), 7.30 (s, 1H), 6.54 (s, 1H), 5.67 (d, J = 4.8 Hz, 1H),5.55 (dd, J = 14.6, 7.3 Hz, 1H), 5.43 (s, 2H), 5.24 (d, J = 19.0 Hz, 1H), 5.03(d, J = 19.0 Hz, 1H), 4.21-4.08 (m, 1H), 3.28 -3.08 (m, 2H), 2.51 (s, 3H), 2.16(d, J = 6.2 Hz, 2H), 1.94-1.82 (m, 2H), 1.42 (d, J = 6.8 Hz, 3H), 0.88 (t, J =7.3 Hz, 3H). ESI-MS (m / z): 524.2 [M+H] + .

[0463] Example 5: (2S)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide and (2S)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide (2R)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide and (2R)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide [ka]

[0464] Step 1: (2S)-2-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropylacetate Mido and (2S)-2-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropylacetamide Furthermore, (2R)-2-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropylacetamide and Synthesis of (2R)-2-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropylacetamide At 25°C, 30.00 mg (61.43 μmol) of 2-23 hydrochloride was dissolved in 1 mL of N,N-dimethylformamide. 2-((tert-butyldiphenylsilyl)oxy)-2-cyclopropylacetic acid (26.13 mg, 73.72 μmol), HATU (35.01 mg, 92.14 μmol), and N,N-diisopropylethylamine (23.82 mg, 184.29 μmol) were added sequentially, and the mixture was reacted at 25°C for 1 hour. The reaction was monitored by LC-MS. Once the reaction was complete, water was added to the reaction solution, and the mixture was extracted with dichloromethane / methanol (volume / volume = 10 / 1). The organic phases were combined, dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified and separated by preparative thin-layer chromatography (dichloromethane:methanol = 15:1) to obtain two isomers. The two isomers were named 2-12-01-A (8.00 mg, Rf value 0.35) and 2-12-01-B (10.00 mg, Rf value 0.40) according to their Rf values.

[0465] Step 2: (2S)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide and (2S)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide Synthesis of (2R)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide and (2R)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide At 25°C, 2-12-01-A (8.00 mg, 10.15 μmol) and 2-12-01-B (10.00 mg, 12.68 μmol) were dissolved in anhydrous THF (1 mL) in two separate reaction flasks. A mixed solution of tetrabutylammonium fluoride (1 M in tetrahydrofuran) / glacial acetic acid (volt / volt = 13 / 1) (50 μL) was added dropwise, and the reaction was allowed to proceed at 25°C for 0.5 hours, with the reaction monitored by LC-MS. Upon completion of the reaction, the reaction solution was purified by preparative high-performance liquid chromatography. 2-12-01-A was used as a starting material and reacted to obtain two isomer products after separation. The corresponding fractions were freeze-dried to obtain compounds 2-12-A (0.77 mg) and 2-12-B (1.03 mg). Simultaneously, 2-12-01-B was used as a starting material and reacted to obtain two isomer products after separation. The corresponding fractions were freeze-dried to obtain compounds 2-12-C (2.50 mg) and 2-12-D (1.00 mg). The purification conditions for 2-12-A / 2-12-B were as follows: Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 8]

[0466] Peak retention time: 2-12-A: 10.0-11.0 minutes, 2-12-B: 11.0-12.5 minutes The purification conditions for 2-12-C / 2-12-D were as follows: Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 9]

[0467] Peak retention time: 2-12-C: 10.6~11.4 minutes, 2-12-D: 11.4~12.5 minutes The structural characteristics data for 2-12-A were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.38 (d, J = 8.9Hz, 1H), 8.15 (s, 1H), 7.30 (s, 1H), 6.54 (s, 1H), 5.63-5.53 (m, 1H), 5.51 (d,J = 5.1 Hz, 1H), 5.42 (s, 2H), 5.28 (d, J = 19.2 Hz, 1H), 5.16 (d, J = 19.1 Hz,1H), 3.60 (t, J = 5.6 Hz, 1H), 3.28-3.11 (m, 2H), 2.52 (s, 3H), 2.22-2.10 (m,2H), 1.86 (tt, J = 14.1, 7.3 Hz, 2H), 1.23 (d, J = 4.9 Hz, 1H), 0.87 (t, J =7.2 Hz, 3H), 0.56-0.36 (m, 4H). ESI-MS (m / z): 550.2 [M+H] + . The structural characteristics data for 2-12-B were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.36 (d, J = 8.7Hz, 1H), 8.16 (s, 1H), 7.31 (s, 1H), 6.55 (s, 1H), 5.57-5.48 (m, 1H), 5.42 (s,2H), 5.40 (d, J = 5.4 Hz, 1H), 5.26 (d, J = 19.3 Hz, 1H), 5.18 (d, J = 19.0 Hz,1H), 3.65-3.60 (m, 1H), 3.26-3.12 (m, 2H), 2.52 (s, 3H), 2.26-2.09 (m, 2H),1.86 (tt, J = 14.1, 7.2 Hz, 2H), 1.19-1.08 (m, 1H), 0.87 (t, J = 7.3 Hz, 3H),0.51-0.27 (m, 4H). ESI-MS (m / z): 550.2 [M+H] + . The structural characteristics data for 2-12-C were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.42 (d, J = 9.0Hz, 1H), 8.15 (s, 1H), 7.31 (s, 1H), 6.54 (s, 1H), 5.56 (dd, J = 14.8, 6.8 Hz,1H), 5.52 (d, J = 5.2 Hz, 1H), 5.42 (s, 2H), 5.29 (d, J = 19.2 Hz, 1H), 5.16(d, J = 19.1 Hz, 1H), 3.62-3.58 (m, 1H), 3.27 -3.08 (m, 2H), 2.51 (s, 3H),2.27-2.08 (m, 2H), 1.87 (tt, J = 14.0, 7.2 Hz, 2H), 1.25 (dd, J = 13.2, 6.8 Hz,1H), 0.87 (t, J = 7.3 Hz, 3H), 0.64-0.28 (m, 4H). ESI-MS (m / z): 550.1 [M+H] + . The structural characteristics data for 2-12-D were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.38 (d, J = 8.7Hz, 1H), 8.16 (s, 1H), 7.31 (s, 1H), 6.54 (s, 1H), 5.59-5.49 (m, 1H), 5.43 (d,J = 5.3 Hz, 1H), 5.43 (s, 2H), 5.26 (d, J = 19.1 Hz, 1H), 5.17 (d, J = 19.0 Hz,1H), 3.64 (t, J = 5.8 Hz, 1H), 3.17 (dd, J = 15.6, 8.3 Hz, 2H), 2.27-2.07 (m,2H), 1.94-1.79 (m, 2H), 1.19-1.06 (m, 1H), 0.87 (t, J = 7.3 Hz, 3H), 0.49-0.28(m, 4H). ESI-MS (m / z): 550.1 [M+H] + .

[0468] Example 6: N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxy-2-methylpropylamine and N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxy-2-methylpropylamine [ka]

[0469] At 25°C, 30.00 mg (61.43 μmol) of 2-23 hydrochloride was dissolved in 1 mL of N,N-dimethylformamide. 16.10 mg (73.72 μmol) of 2-((tert-butyldimethylsilyl)oxy)-2-methylpropionic acid, 35.01 mg (92.14 μmol) of HATU, and 23.82 mg (184.29 μmol) of N,N-diisopropylethylamine were added sequentially, and the reaction was allowed to proceed at 25°C for 1 hour. The reaction was monitored by LC-MS. Upon completion of the reaction, the reaction solution was concentrated, and the crude product was purified and separated by preparative high-performance liquid chromatography to obtain two isomers. The fractions were lyophilized, and the two isomers were named 2-17-A (2.65 mg) and 2-17-B (2.69 mg).

[0470] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 10]

[0471] Peak retention time: 2-17-A: 9.5~10.2 minutes, and 2-17-B: 10.4~10.6 minutes.

[0472] The structural characteristics data for 2-17-A were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.36 (d, J = 9.1Hz, 1H), 8.13 (s, 1H), 7.30 (s, 1H), 6.54 (s, 1H), 5.55-5.45 (m, 2H), 5.42 (s,2H), 5.28 (d, J = 19.0 Hz, 1H), 5.05 (d, J = 19.0 Hz, 1H), 3.27-3.11 (m, 2H),2.51 (s, 1H), 2.24-2.10 (m, 2H), 1.86 (tt, J = 14.0, 7.2 Hz, 2H), 1.46 (s, 3H),1.35 (s, 3H), 0.87 (t, J = 7.3 Hz, 3H). MS m / z (ESI): 538.2 [M+H] + . The structural characteristics data for 2-17-B were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.40 (d, J = 9.2Hz, 1H), 8.13 (s, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.60-5.46 (m, 2H), 5.42 (s,2H), 5.29 (d, J = 19.0 Hz, 1H), 5.02 (d, J = 19.0 Hz, 1H), 3.28 - 3.08 (m, 2H),2.50 (s, 3H), 2.22-2.10 (m, 2H), 1.87 (tt, J = 14.2, 7.2 Hz, 2H), 1.47 (s, 3H),1.35 (s, 3H), 0.88 (t, J = 7.3 Hz, 3H). MS m / z (ESI): 538.2 [M+H] + .

[0473] Example 7: N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-1-hydroxycyclopropane-1-carboxamide and N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-1-hydroxycyclopropane-1-carboxamide [ka]

[0474] Step 1: 1-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)cyclopropane-1-carboxami Synthesis of 1-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)cyclopropane-1-carboxamide At 25°C, 30.00 mg (61.43 μmol) of the hydrochloride salt of 2-23 was dissolved in 1 mL of N,N-dimethylformamide. 1-((tert-butyldiphenylsilyl)oxy)cyclopropane-1-carboxylic acid (25.10 mg, 73.72 μmol), HATU (35.01 mg, 92.14 μmol), and N,N-diisopropylethylamine (23.82 mg, 184.29 μmol) were added sequentially, and the reaction was allowed to proceed at 25°C for 1 hour. The reaction was monitored by LC-MS. Once the reaction was complete, water was added to the reaction solution, and it was extracted with dichloromethane / methanol (volume / volume = 10 / 1). The organic phases were combined, dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was purified and separated by preparative thin-layer chromatography (dichloromethane:methanol = 15:1) to obtain two isomers. The two isomers were named 2-20-01-A (4.00 mg, Rf value 0.30) and 2-20-01-B (4.00 mg, Rf value 0.35) according to their Rf values.

[0475] Step 2: Synthesis of N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-1-hydroxycyclopropane-1-carboxamide and N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-1-hydroxycyclopropane-1-carboxamide At 25°C, 2-20-01-A (4.00 mg, 5.17 μmol) and 2-20-01-B (4.00 mg, 5.17 μmol) were dissolved in anhydrous THF (1 mL) in two reaction flasks, respectively. A mixed solution of tetrabutylammonium fluoride (1 M in tetrahydrofuran) / glacial acetic acid (volt / volt = 13 / 1) (50 μL) was added dropwise, and the reaction was allowed to proceed at 25°C for 0.5 hours, with the reaction monitored by LC-MS. Upon completion of the reaction, the reaction solution was purified by preparative high-performance liquid chromatography, and the fractions were freeze-dried to obtain the title compounds 2-20-A (0.71 mg) and 2-20-B (1.05 mg).

[0476] The purification conditions for 2-20-A were as follows: Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 11]

[0477] The purification conditions for 2-20-B were as follows: Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 12]

[0478] The structural characteristics data for 2-20-A were as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.58 (d, J = 9.0Hz, 1H), 8.15 (s, 1H), 7.31 (s, 1H), 6.55 (s, 1H), 6.30 (s, 1H), 5.55 (dd, J =13.2, 8.2 Hz, 1H), 5.43 (s, 2H), 5.26 (d, J = 19.0 Hz, 1H), 5.10 (d, J = 19.0Hz, 1H), 3.29-3.09 (m, 2H), 2.52 (s, 3H), 2.31-2.15 (m, 2H), 1.93-1.80 (m, 2H),1.25-1.14 (m, 2H), 0.98-0.90 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 536.2 [M+H] + . 2-20-B's structural characteristics are as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.63 (d, J = 9.0Hz, 1H), 8.15 (s, 1H), 7.31 (s, 1H), 6.54 (s, 1H), 6.35 (s, 1H), 5.55 (dd, J =13.5, 8.6 Hz, 1H), 5.43 (s, 2H), 5.29 (d, J = 19.1 Hz, 1H), 5.08 (d, J = 19.1Hz, 1H), 3.28-3.10 (m, 2H), 2.51 (s, 3H), 2.30- 2.14 (m, 2H), 1.93-1.81 (m,2H), 1.26-1.14 (m, 2H), 1.02-0.90 (m, 2H), 0.89 (d, J = 10.9 Hz, 3H). ESI-MS (m / z): 536.2 [M+H] + .

[0479] Example 8: (1S,9S)-1-amino-4-chloro-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione and (1R,9S)-1-amino-4-chloro-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione [ka]

[0480] Step 1: Synthesis of 3-bromo-4-chloro-5-fluoroaniline Compound 3-1-01 (2.00 g, 10.53 mmol) was dissolved in N,N-dimethylformamide (30 mL), and then N-chlorosuccinimide (1.69 g, 12.63 mmol) was slowly added. The reaction was carried out at room temperature for 16 hours and monitored by LC-MS. The reaction solution was concentrated under reduced pressure to obtain the crude product, which was purified by flash silica gel column chromatography (ethyl acetate:petroleum ether = 0-25%) to obtain 0.95 g of the title compound.

[0481] The structural characteristics data was as follows: 1 HNMR (400 MHz, DMSO-d6) δ 6.77 (dd, J = 2.5,1.4 Hz, 1H), 6.51 (dd, J = 11.7, 2.5 Hz, 1H), 5.84 (s, 2H).

[0482] Step 2: Synthesis of N-(3-bromo-4-chloro-5-fluorophenyl)acetamide Compound 3-1-02 (0.95 g, 4.23 mmol) was dissolved in ethyl acetate (20 mL), and acetic anhydride (648.13 mg, 6.35 mmol) was added under nitrogen protection. After the addition, the temperature was raised to 50°C, and the reaction was carried out for 15 hours, monitored by LC-MS. The reaction solution was quenched with methanol (5 mL), and the crude product was directly evaporated to dryness under reduced pressure. This was purified by flash silica gel column chromatography (ethyl acetate:petroleum ether = 0-40%) to obtain 1.01 g of the title compound.

[0483] The structural characteristics data was as follows: ESI-MS (m / z): 265.9[M+H] + .

[0484] Step 3: Synthesis of (E)-4-(5-acetylamino-2-chloro-3-fluorophenyl)-3-butenic acid Compound 3-1-03 and 3-butenoic acid (387.65 mg, 4.50 mmol) were dissolved in a mixed solvent of 1,4-dioxane (24 mL) and water (8 mL). Then, N,N-diisopropylethylamine (1.45 g, 11.26 mmol), tris(o-methylphenyl)phosphine (114.21 mg, 375.24 μmol), and palladium acetate (42.12 mg, 187.62 μmol) were added. After the addition, the reaction system was subjected to nitrogen purging three times, and the reaction was carried out under a nitrogen atmosphere at 100 °C for 16 hours, and the reaction was monitored by LC-MS. After the reaction solution was cooled to room temperature, 1N sodium hydroxide aqueous solution (60 mL) and ethyl acetate (50 mL) were added, and the mixture was shaken to form layers. After separating the aqueous phase, the pH was adjusted to approximately 3 with a 4 mol / L hydrochloric acid aqueous solution, and then extracted with ethyl acetate. The organic phase was combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain 1.00 g of the crude product of the title compound.

[0485] The structural characteristics data was as follows: ESI-MS (m / z): 272.0[M+H] + .

[0486] Step 4: Synthesis of 4-(5-acetylamino-2-chloro-3-fluorophenyl)butanoic acid The crude product of compound 3-1-04 (1.00 g, 3.68 mmol) was dissolved in tetrahydrofuran (15 mL), and then carbon-supported 10% palladium (0.10 g) was added. After the addition, the reaction system was subjected to three substitutions using a hydrogen balloon, and the reaction was carried out under a hydrogen atmosphere for 4 hours and monitored by LC-MS. The reaction solution was filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain 1.00 g of the crude product of the title compound.

[0487] The structural characteristics data was as follows: ESI-MS (m / z): 274.0[M+H] + .

[0488] Step 5: Synthesis of N-(4-chloro-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide The crude product of compound 3-1-05 (1.00 g, 3.65 mmol) was dissolved in trifluoroacetic acid (5 mL), cooled to 5°C, and then trifluoroacetic anhydride (3.84 g, 18.27 mmol, 2.54 mL) was slowly added. The reaction was carried out at 5°C for 2 hours and monitored by LC-MS. The reaction solution was slowly poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. This was purified by flash silica gel column to obtain 0.43 g of the title compound.

[0489] The structural characteristics data was as follows: ESI-MS (m / z): 256.1[M+H] + .

[0490] Step 6: Synthesis of N-(4-chloro-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide Tetrahydrofuran (16 mL) and tert-butanol (4 mL) were added to a reaction flask and cooled to 5°C in an ice bath. Potassium tert-butoxide (415.18 mg, 3.70 mmol) was added, then compound 3-1-06 (0.43 mg, 1.68 mmol) was dissolved in tetrahydrofuran (1 mL) and slowly added. After 10 minutes, isoamyl nitrite (315.24 mg, 2.69 mmol) was added, and the reaction was carried out at 5°C for 1 hour, with the reaction monitored by LC-MS. The reaction solution was quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 455.00 mg of the crude product of the title compound.

[0491] The structural characteristics data was as follows: ESI-MS (m / z): 285.0[M+H] + .

[0492] Step 7: Synthesis of N-(7-amino-4-chloro-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide The crude product of compound 3-1-07 (0.40 g, 1.41 mmol) was dissolved in methanol (10 mL), then 3 mol / L aqueous hydrochloric acid (1 mL) and carbon-supported 10% palladium (40.00 mg) were added. After addition, the reaction system was subjected to three substitutions using a hydrogen balloon, and the reaction was carried out at room temperature under a hydrogen atmosphere for 1 hour, monitored by LC-MS. The reaction solution was filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain 0.43 g of the crude hydrochloride salt of the title compound.

[0493] The structural characteristics data was as follows: ESI-MS (m / z): 271.0[M+H] + .

[0494] Step 8: Synthesis of (9H-fluoren-9-yl)methyl(8-acetamido-5-chloro-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate Crude hydrochloride of compound 3-1-08 (0.43 g, 1.19 mmol) was dissolved in 1,4-dioxane (15 mL), then sodium bicarbonate (400.35 mg, 4.77 mmol), water (5 mL), and 9-fluorenylmethyl-N-succinimidylcarbonate (481.81 mg, 1.43 mmol) were added. After addition, the reaction was carried out at room temperature for 2 hours with stirring and monitored by LC-MS. The reaction solution was poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by C18 reverse-phase column chromatography (acetonitrile: 0.05% in water, formic acid = 20%~100%) to obtain 301.00 mg of the title compound.

[0495] The structural characteristics data was as follows: ESI-MS (m / z): 493.2[M+H] + .

[0496] Step 9: Synthesis of (9H-fluoren-9-yl)methyl(8-amino-5-chloro-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate Compound 3-1-09 (300.00 mg, 608.61 μmol) was dissolved in dioxane (5 mL), 12 mol / L concentrated hydrochloric acid (1 mL) was added, and the temperature was raised to 60°C. The reaction was carried out for 2 hours and monitored by LC-MS. The reaction solution was poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash silica gel column (ethyl acetate:petroleum ether = 0-50%) to obtain 198.00 mg of the title compound.

[0497] The structural characteristics data was as follows: ESI-MS (m / z): 451.1[M+H] + .

[0498] Step 10: Synthesis of (9H-fluoren-9-yl)methyl((9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)carbamate (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolidine-3,6,10(4H)-trione (138.72 mg, 526.96 μmol) and compound 3-1-10 (198.00 mg, 439.13 μmol) were added to toluene (10 mL), then p-toluenesulfonic acid (75.53 mg, 439.13 μmol) was added, and the temperature was raised to 140°C. The reaction was carried out for 4 hours, and the reaction solution was directly evaporated to dryness under reduced pressure at 140°C to obtain the crude product. The crude product was purified by flash silica gel column (methanol:dichloromethane = 0-5%) to obtain 256.00 mg of the title compound.

[0499] The structural characteristics data was as follows: ESI-MS (m / z): 678.1[M+H] + .

[0500] Step 11: Synthesis of (1S,9S)-1-amino-4-chloro-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione and (1R,9S)-1-amino-4-chloro-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione Compound 3-1-11 (201.18 mg, 296.67 μmol) was dissolved in N,N-dimethylformamide (4 mL), then diethylamine (108.49 mg, 1.48 mmol) was added, and the reaction was carried out at room temperature for 0.5 hours, monitored by LC-MS. The reaction solution was removed by vacuum distillation to remove ethylenediamine, and the pH was adjusted to 2-3 with 1 mol / L hydrochloric acid aqueous solution. The reaction solution was then directly purified by preparative high-performance liquid chromatography to obtain the title compounds 3-1-A (44.00 mg) and 3-1-B (43.00 mg).

[0501] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 13]

[0502] 3-1-A (6-minute LCMS, with a retention time of 1.276 minutes, earlier peak) The structural characteristics data was as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.00 (d, J = 10.3Hz, 1H), 7.33 (s, 1H), 6.54 (s, 1H), 5.62 (d, J = 19.3 Hz, 1H), 5.44 (s, 2H),5.38 (d, J = 19.3 Hz, 1H), 4.43-4.38 (m, 1H), 3.28-3.10 (m, 2H), 2.22-2.12 (m,1H), 2.12-2.02 (m, 1H), 1.93-1.80 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 456.1[M+H] + . 3-1-B (6-minute LCMS, with a retention time of 1,300 minutes, later peak) The structural characteristics data were as follows: 1HNMR (400 MHz, DMSO-d6) δ7.98 (d, J = 10.3Hz, 1H), 7.32 (s, 1H), 5.61 (d, J = 19.4 Hz, 1H), 5.44 (s, 2H), 5.32 (d, J =19.4 Hz, 1H), 4.44-4.36 (m, 1H), 3.33-3.25 (m, 1H), 3.22-3.11 (m, 1H), 2.23 -2.13 (m, 1H), 2.11- 2.03 (m, 1H), 1.96 -1.82 (m, 2H), 0.89 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 456.1[M+H] + .

[0503] 6 min LCMS conditions: Chromatography column: Waters SunFire C18 OBD 4.6mm x 50mm x 5.0μm Mobile phase A: 0.05% acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 14]

[0504] Example 9: N-((1S,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide and N-((1R,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide [ka]

[0505] Step 1: Synthesis of (S)-10-benzyl-23-(2-(methylsulfonyl)pyrimidine-5-yl)-6,9,12,15,18-pentoxo-3-oxo-5,8,11,14,17-pentazooctadecane-22-in-carboxylic acid Compound 3-4-01 (30.00 mg, 70.00 μmol) was dissolved in N,N-dimethylformamide (1 mL), and 2,5-dioxopyrrolidine-1-yl-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inylamide (28.00 mg, 77.00 μmol) was added. The reaction was allowed to proceed at room temperature for 1 hour, and the reaction was monitored by LC-MS. The reaction solution was directly purified by preparative high-performance liquid chromatography, and the fraction was freeze-dried to obtain the title compound 3-4-03 (20.00 mg).

[0506] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 15]

[0507] The structural characteristics data was as follows: ESI-MS (m / z): 691.0[M+18]+.

[0508] Step 2: N-((S)-10-benzyl-1-((1S,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inyl-amino Synthesis of N-((S)-10-benzyl-1-((1R,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inyl-amide Compound 3-1-A (36.00 mg, 79.70 μmol) and compound 3-4-03 (64.43 mg, 95.64 μmol) were dissolved in N,N-dimethylformamide (2 mL) in a single stereochemistry. Then, 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholine hydrochloride (46.98 mg, 159.40 μmol) and triethylamine (24.19 mg, 239.10 μmol) were added, and the reaction was carried out at room temperature for 1 hour, monitored by LC-MS. The reaction solution was directly purified by high-performance liquid chromatography to obtain the title compound 3-4-04-A (51.00 mg) in a single stereochemistry.

[0509] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 16]

[0510] The structural characteristics data was as follows: ESI-MS (m / z): 1111.0[M+H] + .

[0511] Compound 3-1-B (36.00 mg, 79.70 μmol) and compound 3-4-03 (64.43 mg, 95.64 μmol) were dissolved in N,N-dimethylformamide (2 mL) in a single stereochemistry. Then, 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholine hydrochloride (46.98 mg, 159.40 μmol) and triethylamine (24.19 mg, 239.10 μmol) were added, and the reaction was carried out at room temperature for 1 hour, monitored by LC-MS. The reaction solution was directly purified by high-performance liquid chromatography to obtain the title compound 3-4-04-B (52.00 mg) in a single stereochemistry.

[0512] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 17]

[0513] The structural characteristics data was as follows: ESI-MS (m / z): 1111.0[M+H] + .

[0514] Step 3: Synthesis of N-((1S,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide and N-((1R,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide Compound 3-4-04-A (40.00 mg, 35.99 μmol) was weighed and dissolved in a mixed solvent of dichloromethane (2 mL) and methanol (1 mL). Then, 4 mol / L ethyl acetate hydrochloride (1 mL) was added, and the reaction was carried out at room temperature for 0.5 hours, monitored by LC-MS. The reaction solution was directly concentrated to dryness under reduced pressure to obtain the crude product. The crude product was purified by high-performance liquid chromatography to obtain the title compound 3-4-A (4.75 mg) in a single stereochemistry.

[0515] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 18]

[0516] The structural characteristics data was as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.50 (d, J = 8.9Hz, 1H), 8.05 (d, J = 10.3 Hz, 1H), 7.33 (s, 1H), 6.55 (s, 1H), 5.67 -5.60 (m,1H), 5.49 (t, J = 5.8 Hz, 1H), 5.43 (s, 2H), 5.21 (s, 2H), 3.96 (d, J = 5.8 Hz,2H), 3.32-3.22 (m, 2H), 2.28-2.15 (m, 2H), 1.93-1.80 (m, 2H), 0.87 (t, J = 7.3Hz, 3H). ESI-MS (m / z): 514.0[M+H] + .

[0517] Compound 3-4-04-B (40.00 mg, 35.99 μmol) was weighed and dissolved in a mixed solvent of dichloromethane (2 mL) and methanol (1 mL). Then, 4 mol / L ethyl acetate hydrochloride (1 mL) was added, and the reaction was carried out at room temperature for 0.5 hours, monitored by LC-MS. The reaction solution was directly concentrated to dryness under reduced pressure to obtain the crude product, and the crude product was purified by high-performance liquid chromatography to obtain the title compound 3-4-B (8.24 mg) in a single stereochemistry.

[0518] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 19]

[0519] The structural characteristics data was as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.52 (d, J = 9.0Hz, 1H), 8.05 (d, J = 10.3 Hz, 1H), 7.34 (s, 1H), 6.55 (s, 1H), 5.68 -5.58 (m,1H), 5.53 (t, J = 5.8 Hz, 1H), 5.43 (d, J = 2.9 Hz, 2H), 5.20 (d, J = 7.3 Hz,2H), 3.97 (d, J = 5.7 Hz, 2H), 3.31-3.21 (m, 2H), 2.26-2.15 (m, 2H), 1.92-1.82(m, 2H), 0.87 (t, J = 7.3 Hz (3H). ESI-MS (m / z): 514.0[M+H] + .

[0520] Example 10: (S)-N-(2-(4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)ethyl)-2-hydroxy-N-isopropylacetamide [ka]

[0521] Step 1: Synthesis of 1-(4-fluoro-3-methylphenyl)-3-(isopropylamino)propan-1-one At 20°C, compound 4-12-01 (500.00 mg, 3.29 mmol), formaldehyde solution (2.5 mL, 37%), and isopropylamine (388.46 mg, 6.57 mmol) were dissolved in isopropanol (5 mL). At 0°C, concentrated hydrochloric acid (2.5 mL) was added dropwise, and the reaction solution was stirred at 100°C for 16 hours and monitored by LC-MS. The reaction solution was concentrated under reduced pressure to obtain the crude product, which was purified by preparative high-performance liquid chromatography, and the fraction was freeze-dried to obtain 200.00 mg of the title compound.

[0522] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 20]

[0523] The structural characteristics data was as follows: ESI-MS (m / z): 224.1 [M+H] + .

[0524] Step 2: Synthesis of 1-(4-fluoro-5-methyl-2-nitrophenyl)-3-(isopropylamino)propan-1-one At 0°C, compound 4-12-02 (100.00 mg, 0.49 mmol) was added to concentrated sulfuric acid (0.5 mL), followed by potassium nitrate (54.34 mg, 0.54 mmol). The reaction mixture was allowed to react at 0°C for 1 hour and monitored by LC-MS. The reaction solution was poured into ice water and purified using a reversed-phase column (acetonitrile: 0.05% formic acid solution = 0-30%) to obtain 90.00 mg of the title compound.

[0525] The structural characteristics data was as follows: ESI-MS (m / z): 269.0 [M+H] + .

[0526] Step 3: Synthesis of 1-(2-amino-4-fluoro-5-methylphenyl)-3-(isopropylamino)propan-1-one At 25°C, compound 4-12-03 (200.00 mg, 0.75 mmol) was added to methanol (20.0 mL), followed by the addition of carbon-supported 10% palladium (10.00 mg). The reaction solution was subjected to hydrogen displacement, and the reaction was carried out under a hydrogen atmosphere at 20°C for 16 hours, with the reaction monitored by LC-MS. The reaction solution was filtered and concentrated under reduced pressure to obtain 183.00 mg of the title compound.

[0527] The structural characteristics data was as follows: ESI-MS (m / z): 239.1 [M+H] + .

[0528] Step 4: Synthesis of (S)-4-ethyl-8-fluoro-4-hydroxy-11-(2-(isopropylamino)ethyl)-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H)-dione At 25°C, compound 4-12-04 (50.00 mg, 0.21 mmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolidine-3,6,10(4H)-trione (55.23 mg, 0.21 mmol) were added to toluene (3 mL), followed by the addition of p-toluenesulfonic acid (3.61 mg, 0.02 mmol). The reaction solution was allowed to react at 130°C for 4 hours and monitored by LC-MS. The reaction solution was concentrated under reduced pressure, the crude product was purified by preparative high-performance liquid chromatography, and the fraction was freeze-dried to obtain 2.00 mg of the trifluoroacetate of the title compound.

[0529] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% trifluoroacetic acid) [Table 21]

[0530] The structural characteristics data was as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.58 (s, 2H), 8.22(d, J = 8.1 Hz, 1H), 7.97 (d, J = 10.7 Hz, 1H), 7.35 (s, 1H), 6.59 (s, 1H),5.47 (s, 2H), 5.41 (s, 2H), 3.58-3.45 (m, 3H), 3.31-3.23 (m, 2H), 2.56 (s, 3H),1.98-1.80 (m, 2H), 1.26 (d, J = 6.3 Hz, 6H), 0.89 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 466.2 [M+H] +

[0531] Step 5: Synthesis of (S)-2-((tert-butyldiphenylsilyl)oxy)-N-(2-(4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)ethyl)-N-isopropylacetamide Compound 4-12-05 (22.00 mg, 47.26 μmol) and 2-((tert-butyldiphenylsilyl)oxy)acetic acid (16.35 mg, 51.99 μmol) were dissolved in N,N-dimethylformamide (1 mL), and then HATU (21.55 mg, 56.71 μmol) and N,N-diisopropylethylamine (18.32 mg, 141.78 μmol) were added. The reaction was carried out at room temperature for 0.5 hours and monitored by LC-MS. The reaction solution was directly purified using a C18 reversed-phase column (acetonitrile: 0.05% formic acid aqueous solution = 30%~100%) to obtain 18.00 mg of the title compound.

[0532] The structural characteristics data was as follows: ESI-MS (m / z): 762.3[M+H] + .

[0533] Step 6: Synthesis of (S)-N-(2-(4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)ethyl)-2-hydroxy-N-isopropylacetamide Compound 4-12-06 (18.00 mg, 23.62 μmol) was dissolved in N,N-dimethylformamide (1 mL), then potassium fluoride (6.86 mg, 118.12 μmol) was added, and the mixture was heated to 50°C and reacted for 1 hour. The reaction was monitored by LC-MS. The reaction solution was directly purified by high-performance liquid chromatography to obtain 1.53 mg of the title compound.

[0534] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 22]

[0535] The structural characteristics data was as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.53 (d, J = 8.2Hz, 1H), 7.91 (d, J = 10.8 Hz, 1H), 7.32 (s, 1H), 6.55 (s, 1H), 5.44 (d, J =13.8 Hz, 4H), 4.72 (t, J = 5.5 Hz, 1H), 4.21 (d, J = 5.5 Hz, 2H), 3.99-3.90 (m,1H), 3.54-3.38 (m, 4H), 2.54 (s, 3H), 1.92-1.83 (m, 2H), 1.17 (dd, J = 6.6, 3.1Hz, 6H), 0.87 (t, J = 7.3 Hz (3H). ESI-MS (m / z): 524.2[M+H] + .

[0536] Example 11: (S)-N-((4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11-yl)methyl)-1-hydroxycyclopropane-carboxamide [ka]

[0537] The raw materials (S)-11-(aminomethyl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1H-pyrano[3',4':6,7]indolidino[1,2-b]quinoline-3,14(4H,12H)-dione (4-10-01, 30.00 mg, 67.00 μmol, prepared according to the method described in International Publication No. 2020219287) and 1-hydroxycyclopropane-carboxylic acid (7.56 mg, 0.074 mmol) were dissolved in DMF (1 mL), and HBTU (34.30 mg, 0.14 mmol) and diisopropylethylamine (26.09 mg, 0.20 mmol) were added while stirring, and the mixture was reacted at room temperature for 4 hours. Water and ethyl acetate were added, the mixture was stirred, and the layers were separated after standing. The organic phase was washed with saturated brine and concentrated under reduced pressure. The concentrate was purified by preparative thin-layer chromatography (dichloromethane:methanol = 20:1), and then by preparative high-performance liquid chromatography to obtain 1.20 mg of solid.

[0538] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 23]

[0539] The structural characteristics data was as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.96 (t, J = 6.0Hz, 1H), 8.51 (d, J = 8.0 Hz, 1H), 7.90 (d, J = 10.8 Hz, 1H), 7.31 (s, 1H),6.53 (s, 1H), 6.30 (s, 2H), 6.30 (s, 1H), 5.52 (s, 2H), 5.44 (s, 2H), 4.84 (d,J = 6.0 Hz, 2H),2.51(s,3H), 1.91-1.81 (m, 2H), 1.01 (dd, J = 7.2, 4.1 Hz, 2H),0.87 (t, J = 7.3 Hz, 3H), 0.83 (t, J = 3.6 Hz, 2H). ESI-MS (m / z): 494.1[M+1]+.

[0540] Example 12: (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione and (1R,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione [ka]

[0541] Step 1: Synthesis of (E)-4-(5-acetylamino-3-fluoro-2-methylphenyl)-2-methyl-3-butenic acid N-(3-bromo-5-fluoro-4-methylphenyl)acetamide (2.00 g, 8.13 mmol) and 2-methyl-3-butenoic acid (976.44 mg, 9.75 mmol) were weighed and dissolved in a mixed solvent of 1,4-dioxane (15 mL) and water (5 mL). Then tris(o-methylphenyl)phosphine (247.37 mg, 812.76 μmol), palladium acetate (91.24 mg, 406.38 μmol), and N,N-diisopropylethylamine (2.31 g, 17.88 mmol) were added. After addition, the reaction system was subjected to nitrogen purging three times, the temperature was raised to 80°C, and the reaction was carried out under a nitrogen atmosphere for 3 hours, monitored by LC-MS. After cooling the reaction solution to room temperature, 1 mol / L aqueous sodium hydroxide solution (60 mL) and ethyl acetate (50 mL) were added, and the layers were separated by shaking. After separating the lower layer of the aqueous phase, the pH was adjusted to approximately 3 with a 4 mol / L hydrochloric acid aqueous solution, followed by extraction with ethyl acetate. The organic phase was then combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 1.90 g of the title compound.

[0542] The structural characteristics data was as follows: ESI-MS (m / z): 266.1[M+H] + .

[0543] Step 2: Synthesis of 4-(5-acetylamino-3-fluoro-2-methylphenyl)-2-methylbutanoic acid (E)-4-(5-acetylamino-3-fluoro-2-methylphenyl)-2-methyl-3-butenoic acid (1.90 g, 7.16 mmol) was dissolved in methanol (40 mL), and 10% carbon-supported palladium (0.15 g) was added under nitrogen protection. The reaction system was then subjected to three substitutions using a hydrogen balloon, and the reaction was carried out under a hydrogen atmosphere for 2 hours and monitored by LC-MS. The reaction solution was filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain 1.51 g of the title compound.

[0544] The structural characteristics data was as follows: ESI-MS (m / z): 268.1[M+H] + .

[0545] Step 3: Synthesis of N-(3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 4-(5-acetylamino-3-fluoro-2-methylphenyl)-2-methylbutanoic acid (1.50 g, 5.61 mmol) was weighed, dissolved in trifluoroacetic acid (20 mL), cooled to 5°C, and then trifluoroacetic anhydride (2.36 g, 11.22 mmol) was added dropwise. After the addition, the reaction was carried out at 5°C for 2 hours and monitored by LC-MS. The reaction solution was slowly poured into saturated sodium bicarbonate aqueous solution, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. The crude product was purified by flash silica gel column (ethyl acetate:petroleum ether = 0-30%) to obtain 1.05 g of the title compound.

[0546] The structural characteristics data was as follows: ESI-MS (m / z): 250.1[M+H] + .

[0547] Step 4: Synthesis of N-(7-bromo-3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide N-(3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (0.55 g, 2.21 mmol) was weighed and dissolved in acetic acid (8 mL). Bromine (387.85 mg, 2.43 mmol) was then added, and the temperature was raised to 50°C. The reaction was carried out for 15 hours and monitored by LC-MS. The reaction solution was directly evaporated to dryness under reduced pressure to obtain the crude product, which was purified by flash silica gel column chromatography (ethyl acetate:petroleum ether = 0-30%) to obtain 461.00 mg of the title compound.

[0548] The structural characteristics data was as follows: ESI-MS (m / z): 328.0[M+H] + .

[0549] Step 5: Synthesis of N-(7-azido-3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide N-(7-bromo-3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (460.00 mg, 1.40 mmol) was weighed and dissolved in N,N-dimethylformamide (10 mL). Then sodium azide (273.37 mg, 4.21 mmol) was added, and the reaction was carried out at room temperature for 1 hour and monitored by LC-MS. The reaction solution was slowly poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. This was purified by flash silica gel column (ethyl acetate:petroleum ether = 0-50%) to obtain 347.00 mg of the title compound.

[0550] The structural characteristics data was as follows: ESI-MS (m / z): 291.1[M+H] + .

[0551] Step 6: Synthesis of N-(7-amino-3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide N-(7-azido-3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (347.00 mg, 1.20 mmol) was weighed and dissolved in tetrahydrofuran (10 mL). Under nitrogen protection, 10% carbon-supported palladium (30.00 mg) was added, and the reaction system was subjected to three substitutions using a hydrogen balloon. The reaction was carried out under a hydrogen atmosphere for 2 hours and monitored by LC-MS. The reaction solution was filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain the crude product. This was purified using a C18 reverse-phase column (acetonitrile: 0.05% formic acid aqueous solution = 0%~30%) to obtain 205.00 mg of the title compound.

[0552] The structural characteristics data was as follows: ESI-MS (m / z): 265.1[M+H] + .

[0553] Step 7: Synthesis of (9H-fluoren-9-yl)methyl(8-acetylamino-6-fluoro-2,5-dimethyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate N-(7-amino-3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide (200.00 mg, 756.73 μmol) was weighed and dissolved in a mixed solvent of 1,4-dioxane (6 mL) and water (3 mL). Then sodium bicarbonate (254.28 mg, 3.03 mmol) and 9-fluorenylmethyl-N-succinimide carbonate (650.55 mg, 1.14 mmol) were added, and the reaction was carried out at room temperature for 2 hours with stirring, and monitored by LC-MS. The reaction solution was slowly poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. This was purified using a C18 reversed-phase column (acetonitrile: 0.05% formic acid aqueous solution = 20%~80%) to obtain 301.00 mg of the title compound.

[0554] The structural characteristics data was as follows: ESI-MS (m / z): 487.0[M+H] + .

[0555] Step 8: Synthesis of (9H-fluoren-9-yl)methyl(8-amino-6-fluoro-2,5-dimethyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate (9H-fluoren-9-yl)methyl(8-acetylamino-6-fluoro-2,5-dimethyl-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate (101.00 mg, 207.59 μmol) was dissolved in 1,4-dioxane (5 mL), then 3 mol / L hydrochloric acid aqueous solution (5 mL) was added, the temperature was raised to 50°C, and the reaction was carried out for 15 hours and monitored by LC-MS. The reaction solution was slowly poured into saturated sodium bicarbonate aqueous solution, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. This was purified by flash silica gel column (methanol:dichloromethane = 0%~5%) to obtain 71.00 mg of the title compound.

[0556] The structural characteristics data was as follows: ESI-MS (m / z): 445.2[M+H] + .

[0557] Step 9: Synthesis of (9H-fluoren-9-yl)methyl((9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)carbamate (9H-fluoren-9-yl)methyl(8-amino-6-fluoro-2,5-dimethyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate (35.00 mg, 132.96 μmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indrazine-3,6,10(4H)-trione (49.25 mg, 110.80 μmol) were added to toluene (3 mL), followed by the addition of p-toluenesulfonic acid (19.08 mg, 110.80 μmol). The temperature was raised to 140 °C, and the reaction was carried out for 4 hours. The liquid reaction solution was directly evaporated to dryness under reduced pressure at 140 °C to obtain the crude product. The crude product was purified using a C18 reversed-phase column (acetonitrile: 0.05% formic acid aqueous solution = 20%~80%) to obtain 21.00 mg of the title compound.

[0558] The structural characteristics data was as follows: ESI-MS (m / z): 672.2[M+H] + .

[0559] Step 10: Synthesis of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione and (1R,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione (9H-fluoren-9-yl)methyl((9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)carbamate (21.00 mg, 31.26 μmol) was dissolved in N,N-dimethylformamide (1 mL), then diethylamine (0.2 mL) was added, and the reaction was carried out at room temperature for 0.5 hours and monitored by LC-MS. The reaction solution was distilled under reduced pressure to remove ethylenediamine, and then the pH was adjusted to 2-3 with a 1 mol / L aqueous hydrochloric acid solution. The reaction solution was then directly purified by preparative high-performance liquid chromatography to obtain two isomers, 5-13-A (1.30 mg) and 5-13-B (1.68 mg).

[0560] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 24]

[0561] The structural characteristics data for 5-13-A (6-minute LCMS, with an earlier peak and retention time of 1.373 minutes) were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 7.88 (d, J = 10.6Hz, 1H), 7.36 (s, 1H), 6.58 (s, 1H), 5.60 (d, J = 3.5 Hz, 2H), 5.46 (d, J = 2.5Hz, 2H), 3.25-3.17 (m, 2H), 2.41 (s, 3H), 2.38-2.28 (m, 2H), 1.91-1.84 (m, 2H),1.79 (s, 3H)), 0.88 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 450.2[M+H] + . The structural characteristics data for 5-13-B (6-minute LCMS, with a later peak having a retention time of 1.523 minutes) were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 7.76 (d, J = 10.8Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.73 (d, J = 19.8 Hz, 1H), 5.50 -5.40 (m,3H), 3.26-3.17 (m, 1H), 3.08-2.96 (m, 1H), 2.38 (s, 3H), 2.19-2.11 (m, 1H),2.04 (td, J = 13.0, 5.1 Hz, 1H), 1.91-1.79 (m, 2H), 1.34 (s, 3H), 0.87 (t, J =7.3 Hz, 3H). ESI-MS (m / z): 450.2[M+H] + .

[0562] 6 min LCMS conditions: Chromatography column: Waters SunFire C18 OBD 4.6mm x 50mm x 5.0μm Mobile phase A: 0.05% acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 25]

[0563] Example 13: (1S,9S)-1-(aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione and (1R,9S)-1-(aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione [ka]

[0564] Step 1: Synthesis of N-(7-((dimethylamino)methylene)-3-fluoro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide Compound 5-13-04 (1.00 g, 4.25 mmol) was dissolved in N,N-dimethylformamide dimethylacetal (10 mL), then heated to 120°C and reacted for 3 hours, with the reaction monitored by LC-MS. After cooling the reaction solution to room temperature, it was directly evaporated to dryness under reduced pressure to obtain the crude product, which was purified by flash silica gel column chromatography (ethyl acetate:petroleum ether = 20%~100%) to obtain 891.00 mg of the title compound.

[0565] The structural characteristics data was as follows: ESI-MS (m / z): 291.1[M+H] + .

[0566] Step 2: Synthesis of N-(7-(aminomethylene)-3-fluoro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide Compound 5-7-01 (0.89 g, 3.07 mmol) was dissolved in ethanol (25 mL), then ammonium acetate (2.36 g, 30.65 mmol) was added, and the reaction was carried out at room temperature for 16 hours, monitored by LC-MS. The reaction solution was evaporated to dryness under reduced pressure, then dichloromethane (30 mL) and water (20 mL) were added, the mixture was stirred, and the organic phase was separated by standing. The mixture was dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain 785.00 mg of the title compound.

[0567] The structural characteristics data was as follows: ESI-MS (m / z): 263.1[M+H] + .

[0568] Step 3: Synthesis of N-(7-(aminomethyl)-3-fluoro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide Compound 5-7-02 (0.80 g, 3.05 mmol) was dissolved in ethanol (200 mL), then carbon-supported 10% palladium (0.40 mg) and concentrated hydrochloric acid (0.2 mL) were added. The reaction system was subjected to three substitutions using a hydrogen balloon, and the reaction was carried out at room temperature under a hydrogen atmosphere for 3 hours, monitored by LC-MS. The reaction solution was filtered directly, and the filtrate was evaporated to dryness under reduced pressure to obtain 905.00 mg of the hydrochloride salt of the title compound.

[0569] The structural characteristics data was as follows: ESI-MS (m / z): 265.1[M+H] + .

[0570] Step 4: Synthesis of (9H-fluoren-9-yl)methyl((8-acetamido-6-fluoro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)methyl)carbamate 0.90 g, 2.99 mmol of the hydrochloride salt of compound 5-7-03 was dissolved in 20 mL of 1,4-dioxane. Then, sodium bicarbonate (1.01 g, 11.97 mmol), water (10 mL), and 9-fluorenylmethyl-N-succinimidylcarbonate (1.21 g, 3.59 mmol) were added. After addition, the reaction was carried out at room temperature for 1 hour with stirring and monitored by LC-MS. The reaction solution was poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure to obtain the crude product. The crude product was purified by flash silica gel column chromatography (ethyl acetate:petroleum ether = 0-50%) to obtain 1.30 g of the title compound.

[0571] The structural characteristics data was as follows: ESI-MS (m / z): 487.1[M+H] + .

[0572] Step 5: Synthesis of (9H-fluoren-9-yl)methyl((8-amino-6-fluoro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)methyl)carbamate Compound 5-7-04 (0.80 g, 1.64 mmol) was dissolved in 1,4-dioxane (20 mL), and 3 mol / L hydrochloric acid aqueous solution (20 mL) was added under nitrogen protection. After addition, the temperature was raised to 60°C, and the reaction was carried out for 15 hours and monitored by LC-MS. The reaction solution was slowly poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. The crude product was purified by flash silica gel column (ethyl acetate:petroleum ether = 0-40%) to obtain 561.00 mg of the title compound.

[0573] The structural characteristics data was as follows: ESI-MS (m / z): 445.1[M+H] + .

[0574] Step 6: Synthesis of (9H-fluoren-9-yl)methyl(((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]indolidino[1,2-b]quinoline-1-yl)methyl)carbamate (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolidine-3,6,10(4H)-trione (597.00 mg, 2.27 mmol) and compound 5-7-05 (840.00 mg, 1.89 mmol) were added to toluene (60 mL), followed by the addition of p-toluenesulfonic acid (325.00 mg, 1.89 mmol). After the addition, the temperature was raised to 140°C and the reaction was carried out for 4 hours. The reaction solution was then directly evaporated to dryness under reduced pressure at 140°C to obtain the crude product. The crude product was purified by flash silica gel column (methanol:dichloromethane = 0-5%) to obtain 563.00 mg of the title compound.

[0575] The structural characteristics data was as follows: ESI-MS (m / z): 672.2[M+H] + .

[0576] Step 7: Synthesis of (1S,9S)-1-(aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione and (1R,9S)-1-(aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione Compound 5-7-06 (454.00 mg, 675.89 μmol) was dissolved in N,N-dimethylformamide (5 mL), then diethylamine (1 mL) was added, and the reaction was allowed to proceed at room temperature for 0.5 hours. The reaction was monitored by LC-MS. After evaporating ethylenediamine from the reaction solution under reduced pressure, the pH was adjusted to 2-3 with formic acid, and the reaction solution was directly purified by preparative high-performance liquid chromatography. The fractions were freeze-dried to obtain the title compounds 5-7-A (32.00 mg) and 5-7-B (56.00 mg).

[0577] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 26]

[0578] The structural characteristics data for 5-7-A (6-minute LCMS, with an earlier peak and retention time of 1.488 minutes) were as follows: 1HNMR (400 MHz, DMSO-d6) δ 7.88 (d, J = 10.6Hz, 1H), 7.36 (s, 1H), 6.58 (s, 1H), 5.60 (d, J = 3.5 Hz, 2H), 5.46 (d, J = 2.5Hz, 2H), 3.25-3.17 (m, 2H), 2.41 (s, 3H), 2.38-2.28 (m, 2H), 1.91-1.84 (m, 2H),1.79 (s, 3H)), 0.88 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 450.2[M+H] + . The structural characteristics data for 5-7-B (6-minute LCMS, with a later peak having a retention time of 1.596 minutes) were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 7.76 (d, J = 10.8Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.73 (d, J = 19.8 Hz, 1H), 5.50 -5.40 (m,3H), 3.26-3.17 (m, 1H), 3.08-2.96 (m, 1H), 2.38 (s, 3H), 2.19-2.11 (m, 1H),2.04 (td, J = 13.0, 5.1 Hz, 1H), 1.91- 1.79 (m, 2H), 1.34 (s, 3H), 0.87 (t, J =7.3 Hz, 3H). ESI-MS (m / z): 450.2[M+H] + .

[0579] 6 min LCMS conditions: Chromatography column: Waters SunFire C18 OBD 4.6mm x 50mm x 5.0μm Mobile phase A: 0.05% acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 27]

[0580] Example 14: N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-1-hydroxycyclopropane-1-carboxamide or N-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-1-hydroxycyclopropane-1-carboxamide [ka]

[0581] Step 1: 1-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)cyclopropane-1-carboxami Synthesis of 1-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)cyclopropane-1-carboxamide Compound 5-13-A (10.00 mg, 22.25 μmol) was dissolved in a single stereochemistry and 1-((tert-butyldiphenylsilyl)oxy)cyclopropane-1-carboxylic acid (11.36 mg, 33.37 μmol) was dissolved in N,N-dimethylformamide (1 mL). Then, HATU (12.68 mg, 33.37 μmol) and N,N-diisopropylethylamine (8.63 mg, 66.74 μmol) were added, and the reaction was carried out at room temperature for 0.5 hours, monitored by LC-MS. The reaction solution was directly purified using a C18 reversed-phase column (acetonitrile: 0.05% formic acid aqueous solution = 30%~100%) to obtain the title compound 5-16-01-A (7 mg) in a single stereochemistry.

[0582] The structural characteristics data was as follows: ESI-MS (m / z): 772.3[M+H] + .

[0583] Step 2: Synthesis of N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-1-hydroxycyclopropane-1-carboxamide or N-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-1-hydroxycyclopropane-1-carboxamide Compound 5-16-01-A (7.00 mg, 9.07 μmol) was dissolved in N,N-dimethylformamide (1 mL), then potassium fluoride (2.63 mg, 45.34 μmol) was added, and the temperature was raised to 50°C. The reaction was carried out for 1 hour and monitored by LC-MS. The reaction solution was directly purified by high-performance liquid chromatography to obtain the single stereoisomer of the title compound 5-16-A (1.73 mg).

[0584] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 28]

[0585] The structural characteristics data was as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.26 (s, 1H), 7.78(d, J = 10.8 Hz, 1H), 7.30 (s, 1H), 6.56 (s, 1H), 6.52 (s, 1H), 5.52 (d, J =19.3 Hz, 1H), 5.43 (d, J = 4.4 Hz, 2H), 4.94 (d, J = 19.2 Hz, 1H), 3.30-3.24(m,1H), 3.11-3.00 (m, 1H), 2.95-2.84 (m, 1H), 2.39 (s, 3H), 1.98-1.80 (m, 3H),1.62 (s, 3H), 0.87 (t, J = 7.9 Hz, 3H). ESI-MS (m / z): 534.2[M+H] + .

[0586] Example 15: N-((10S)-10-benzyl-1-(((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4';6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexadecane Amide or N-((10S)-10-benzyl-1-(((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4';6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexadecanamide [ka]

[0587] Step 1: Separation and purification of (9S)-1-amino-5-chloro-9-ethyl-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzopyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione Compound 2-23 (16.00 mg) was purified by preparative high-performance liquid chromatography, and two diastereoisomers were separated under the following purification conditions to obtain 5.10 mg of trifluoroacetate of 2-23-A (retention time 9.85 min) and 7.12 mg of trifluoroacetate of 2-23-B (retention time 10.62 min).

[0588] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% trifluoroacetic acid) Table 29

[0589] Structural characteristics: 2-23-A: 1 HNMR (400 MHz, DMSO-d6) δ 8.42 (s, 3H), 8.27 (s, 1H), 7.36 (s, 1H), 6.59 (s, 1H), 5.78-5.63 (m, 1H), 5.50- 5.36 (m, 3H),5.10-5.06 (m, 1H), 3.20-3.04 (m, 2H), 2.56 (s, 3H), 2.26-2.13 (m, 2H), 1.93-1.79 (m, 2H), 0.88 (t, J = 7.2 Hz, 3H). ESI-MS (m / z): 452.1 [M+H] + . 2-23-B: 1 HNMR (400 MHz, DMSO-d6) δ 8.42 (s, 3H), 8.27 (s, 1H), 7.36 (s, 1H), 6.58 (s, 1H), 5.78-5.63 (m, 1H), 5.50- 5.36 (m, 3H),5.10-5.06 (m, 1H), 3.20-3.04 (m, 2H), 2.55 (s, 3H), 2.26-2.13 (m, 2H), 1.93-1.79 (m, 2H), 0.88 (t, J = 7.2 Hz, 3H). ESI-MS (m / z): 452.0 [M+H] + .

[0590] Step 2: N-((10S)-10-benzyl-1-(((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4';6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexadecane Synthesis of N-((10S)-10-benzyl-1-(((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4';6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexadecanamide At 25°C, trifluoroacetate of 2-23-A (34.71 mg, 61.43 μmol) was dissolved in N,N-dimethylformamide (1 mL), and 3-4-03 (49.66 mg, 73.72 μmol), HATU (35.01 mg, 92.14 μmol), and N,N-diisopropylethylamine (23.82 mg, 184.29 μmol) were added sequentially. The reaction was allowed to proceed at 25°C for 0.5 hours, and the reaction was monitored by LC-MS. Once the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (under the following conditions), and the fraction was freeze-dried to obtain the title compound DL-15 (11.04 mg) with a retention time of 7.5 minutes.

[0591] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 30]

[0592] The structural characteristics data was as follows: DL-15: ESI-MS (m / z): 1107.3[M+H] + .

[0593] Example 16: N-((9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide [ka]

[0594] Step 1: Synthesis of N-((9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-cyclopropyl-2-hydroxyacetamide Formate (50 mg, 109.68 μmol) of the single stereoisomer compound 3-1-A and 2-cyclopropyl-2-hydroxyacetic acid (25.47 mg, 219.36 μmol) were dissolved in N,N-dimethylformamide (2 mL). Then, HATU (7.57 mg, 219.36 μmol) and N,N-diisopropylethylamine (42.53 mg, 329.04 μmol) were added, and the reaction was carried out at room temperature for 0.5 hours. The reaction was monitored by LC-MS, and the reaction solution was directly purified by preparative high-performance liquid chromatography to obtain two isomers of the title compound (3-12-A: 12.96 mg, 3-12-B: 13.56 mg).

[0595] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 31]

[0596] The structural characteristics data for 3-12-A (6-minute LCMS, earlier peak) were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.44 (d, J = 9.0Hz, 1H), 8.05 (d, J = 10.2 Hz, 1H), 7.33 (s, 1H), 6.54 (s, 1H), 5.62 (q, J =6.7 Hz, 1H), 5.52 (d, J = 5.1 Hz, 1H), 5.42 (s, 2H), 5.24 (q, J = 19.2 Hz, 2H),3.61 (dd, J = 6.2, 5.1 Hz, 1H), 3.32 - 3.21 (m, 2H), 2.19 (q, J = 6.5 Hz, 2H),1.92 - 1.80 (m, 2H), 1.26 - 1.20 (m, 1H), 0.87 (t, J = 7.3 Hz, 3H), 0.57 - 0.34(m, 4H). ESI-MS (m / z): 554.0 [M+H] + . The structural characteristics data for 3-12-B (6-minute LCMS, later peak) were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.43 (d, J = 8.7Hz, 1H), 8.06 (d, J = 10.3 Hz, 1H), 7.33 (s, 1H), 5.57 (q, J = 6.7 Hz, 1H),5.43 (s, 2H), 5.30 - 5.17 (m, 2H), 3.64 (d, J = 6.2 Hz, 1H), 3.29 (q, J = 6.7Hz, 2H), 2.28 - 2.13 (m, 2H), 1.93 - 1.78 (m, 2H), 1.18 - 1.08 (m, 1H), 0.87(t, J = 7.3 Hz, 3H), 0.50 - 0.29 (m, 4H). ESI-MS (m / z): 554.0 [M+H] +.

[0597] The formate (50 mg, 109.68 μmol) and 2-cyclopropyl-2-hydroxyacetic acid (25.47 mg, 219.36 μmol) of the other single stereoisomer compound, 3-1-B, were dissolved in N,N-dimethylformamide (2 mL). Then, HATU (7.57 mg, 219.36 μmol) and N,N-diisopropylethylamine (42.53 mg, 329.04 μmol) were added, and the reaction was carried out at room temperature for 0.5 hours. The reaction was monitored by LC-MS, and the reaction solution was directly purified by preparative high-performance liquid chromatography to obtain two isomers of the title compound (3-12-C: 20.19 mg, 3-12-D: 18.33 mg).

[0598] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 32]

[0599] The structural characteristics data for 3-12-C (6-minute LCMS, earlier peak) were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.47 (d, J = 9.0Hz, 1H), 8.06 (d, J = 10.3 Hz, 1H), 7.33 (s, 1H), 6.54 (s, 1H), 5.62 (q, J =6.5 Hz, 1H), 5.53 (d, J = 5.1 Hz, 1H), 5.43 (s, 2H), 5.32 - 5.16 (m, 2H), 3.61(dd, J = 6.3, 5.1 Hz, 1H), 3.32 - 3.22 (m, 2H), 2.19 (q, J = 6.5 Hz, 2H), 1.92- 1.80 (m, 2H), 1.28 - 1.20 (m, 1H), 0.87 (t, J = 7.3 Hz, 3H), 0.54 - 0.35 (m,4H). ESI-MS (m / z): 554.0 [M+H] + . The structural characteristics data for 3-12-D (6-minute LCMS, later peak) were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.44 (d, J = 8.8 Hz,1H), 8.05 (d, J = 10.2 Hz, 1H), 7.34 (s, 1H), 6.55 (s, 1H), 5.58 (q, J = 6.7Hz, 1H), 5.45 (d, J = 5.2 Hz, 1H), 5.43 (s, 2H), 5.31 - 5.14 (m, 2H), 3.65 (t,J = 5.7 Hz, 1H), 3.33 - 3.21 (m, 2H), 2.28 - 2.13 (m, 2H), 1.95 - 1.80 (m, 2H),1.16 - 1.09 (m, 1H), 0.88 (t, J = 7.3 Hz, 3H), 0.46 - 0.31 (m, 4H). ESI-MS (m / z): 554.0 [M+H] + .

[0600] Example 17: Preparation of (S)-N-((1S,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine and (S)-N-((1R,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine [ka]

[0601] At 25°C, (9S)-1-amino-4-chloro-9-ethyl-5-fluoro-9-hydroxy-2,3,12,15-tetrahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13(1H,9H)-dione (80.0 mg, 175.5 μmol) and L-lactic acid (31.6 mg, 351.0 μmol) were dissolved in DMF (3 mL), then HATU (121.1 mg, 351.0 μmol) and DIPEA (68.0 mg, 526.5 μmol) were added, and the mixture was reacted at room temperature for 2 hours. The reaction solution was directly purified by preparative high-performance liquid chromatography to obtain compound 3-7-A (6.1 mg, yield 12%) and compound 3-7-B (9.6 mg, yield 20%).

[0602] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 33]

[0603] The structural characteristics data for compound 3-7-A (6 min LC-MS, earlier peak with retention time of 2.49 min) were as follows: MS m / z (ESI): 528.2 [M+H] + 1HNMR (400 MHz, DMSO-d6) δ 8.55 (d, J = 9.2Hz, 1H), 8.06 (d, J = 10.4 Hz, 1H), 7.33 (s, 1H), 6.55 (s, 1H), 6.67 (d, J =4.8 Hz, 1H), 5.65 - 5.59 (m, 1H), 5.43 (s, 2H), 5.29 - 5.21 (m, 1H), 5.14 -5.10 (m, 1H), 4.15 - 4.10 (m,, 1H), 3.27 - 3.20 (m, 1H), 2.22 - 2.15 (m, 2H),1.92 - 1.81 (m, 2H), 1.41 (d, J = 6.8 Hz, 3H), 1.30 - 1.23 (m, 1H), 0.89 - 0.85(t, J = 7.2 Hz, 3H). The structural characteristics data for compound 3-7-B (later peak with retention time of 2.50 minutes, measured at 6 min LC-MS) were as follows: MS m / z (ESI): 528.2 [M+H] + 1 HNMR (400 MHz, DMSO-d6) δ 8.46 (d, J = 8.8Hz, 1H), 8.06 (d, J = 10.4 Hz, 1H), 7.33 (s, 1H), 6.56 (s, 1H), 5.60 - 5.53(ms, 1H), 5.51 (d, J = 5.2 Hz, 1H), 5.43 (s, 2H), 5.27 - 5.14 (m, 2H), 4.16 -4.08 (m, 1H), 3.28 - 3.22 (m, 1H), 2.22 - 2.19 (m, 2H), 1.92 - 1.81 (m, 2H),1.49 - 1.39 (m, 1H), 1.29 (d, J = 6.8 Hz, 3H), 0.87 (t, J = 7.2 Hz, 3H).

[0604] Example 18: Synthesis of N-((1S,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl-1-hydroxycyclopropylcarboxamide and N-((1R,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl-1-hydroxycyclopropylcarboxamide [ka]

[0605] At 25°C, (9S)-1-amino-4-chloro-9-ethyl-5-fluoro-9-hydroxy-2,3,12,15-tetrahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13(1H,9H)-dione (80 mg, 175.49 μmol) and 1-hydroxycyclopropanecarboxylic acid (35.83 mg, 350.98 μmol) were heated in DMF. The compound was dissolved in (2 mL), then HATU (121.14 mg, 350.98 μmol) and DIPEA (68.04 mg, 526.47 μmol) were added. The reaction was carried out at room temperature for 0.5 hours, monitored by HPLC-MS, and the reaction solution was concentrated to dryness. It was then purified directly by preparative high-performance liquid chromatography to obtain the title compounds 3-17-A (5.3 mg) and 3-17-B (3.5 mg).

[0606] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 34]

[0607] The structural characteristics data was as follows: The structural characteristics data for 3-17-A (6-minute LCMS, with an earlier peak and retention time of 2.657 minutes) were as follows: ESI-MS (m / z): 540.0 [M+H] + . 1 HNMR (400 MHz, DMSO-d6) δ 8.69 (d, J = 8.8Hz, 1H), 8.06 (d, J = 10.2 Hz, 1H), 7.35 (s, 1H), 6.57 (s, 1H), 6.32 (s, 1H),5.62 (s, 1H), 5.45 (s, 2H), 5.34 - 5.24 (m, 1H), 5.20 - 5.10(m, 1H), 2.26 (s,2H), 2.00 (s, 1H), 1.88 (s, 2H), 1.47 - 1.12 (m, 8H), 1.01 - 0.80(m, 6H). The structural characteristics data for 3-17-A (6-minute LCMS, with a later peak and retention time of 2.724 minutes) were as follows: ESI-MS (m / z): 540.0 [M+H] + . 1 HNMR (400 MHz, DMSO-d6) δ 8.69 (d, J = 8.8 Hz,1H), 8.06 (d, J = 10.2 Hz, 1H), 7.35 (s, 1H), 6.57 (s, 1H), 6.32 (s, 1H), 5.62(s, 1H), 5.45 (s, 2H), 5.34 - 5.24 (m, 1H), 5.20 - 5.10(m, 1H), 2.26 (s, 2H),2.00 (s, 1H), 1.88 (s, 2H), 1.47 - 1.12 (m, 8H), 1.01 - 0.80(m, 6H).

[0608] Example 19: Synthesis of N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-1,2,3,9,10,12,13,15-tetrahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide [ka]

[0609] Step 1: Synthesis of N-(4-chloro-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide N-(4-chloro-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide (570 mg, 2.23 mmol) and cyclopropylboronic acid (574.56 mg, 6.69 mmol) were dissolved in 1,4-dioxane, and then dichlorodi-tert-butyl-(4-dimethylaminophenyl)phosphine palladium(II) (480 mg, 677.97 μmol) and cesium carbonate (2.17 g, 6.69 mmol) were added. l) was added, and the reaction was carried out under nitrogen protection at 115°C in microwaves for 2 hours, monitored by LC-MS, diluted with ethyl acetate, filtered, extracted with ethyl acetate (30 ml x 3), combined with the organic phase, washed with saturated brine (50 mL), dehydrated with anhydrous sodium sulfate, filtered, evaporated to dryness under reduced pressure, and the resulting crude product was purified by column chromatography using a silica gel column (PE:EA = 1:4) to obtain 550 mg of the title compound.

[0610] The structural characteristics data was as follows: ESI-MS (m / z): 262.1[M+H] + .

[0611] Step 2: Synthesis of N-(4-cyclopropyl-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide Tetrahydrofuran (30 mL) and tert-butanol (10 mL) were added to the reaction flask and cooled to 5°C in an ice bath. Then potassium tert-butoxide (945 mg, 8.42 mmol) was added, followed by the dissolution of N-(4-chloro-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (1.0 g, 3.83 mmol) in tetrahydrofuran (1 mL), and the mixture was added dropwise. The solution was slowly added by method, followed by the addition of isoamyl nitrite (718 mg, 6.12 mmol) 10 minutes later. After addition, the reaction was carried out at 5°C for 1 hour and monitored by LC-MS. The reaction solution was quenched with saturated ammonium chloride aqueous solution (50 mL), extracted with ethyl acetate (40 ml x 3), the organic phases were combined, washed with saturated brine (50 mL), dehydrated with anhydrous sodium sulfate, then filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain 1.2 g of the crude product of the title compound.

[0612] The structural characteristics data was as follows: ESI-MS (m / z): 291.1[M+H] + .

[0613] Step 3: Synthesis of N-(7-amino-4-cyclopropyl-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide hydrochloride Crude product (1.2 g, 1.41 mmol) of N-(4-cyclopropyl-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide was dissolved in methanol (7.5 mL) and tetrahydrofuran (7.5 mL). Then, 1 mol / L aqueous hydrochloric acid (7.5 mL) and carbon-supported 10% palladium (450 mg) were added. After addition, the reaction system was subjected to three substitutions using a hydrogen balloon. The reaction was carried out at room temperature under a hydrogen atmosphere for 1 hour, monitored by LC-MS, the reaction solution was filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain 1.05 g of crude product.

[0614] The structural characteristics data was as follows: ESI-MS (m / z): 277.1[M+H]+ .

[0615] Step 4: Synthesis of (9H-fluoren-9-yl)methyl(8-acetamido-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate The crude product of N-(7-amino-4-cyclopropyl-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide hydrochloride (1.05 g, 3.80 mmol) was dissolved in 1,4-dioxane (10 mL), then sodium bicarbonate (1.3 g, 15.20 mmol), water (10 mL), and 9-fluorenylmethyl-N-succinimidylcarbonate (1.54 g, 4.56 mmol) were added. The reaction was carried out at room temperature for 2 hours with stirring, monitored by LC-MS, and the reaction solution was poured into water (50 mL). Then extracted with ethyl acetate (40 mL x 3), the organic phases were combined, washed with saturated brine (50 mL), dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure to obtain the crude product. The crude product was purified by C18 reverse-phase column chromatography to obtain 2.0 g of the title compound.

[0616] The structural characteristics data was as follows: ESI-MS (m / z): 499.2[M+H] + .

[0617] Step 5: Synthesis of (9H-fluoren-9-yl)methyl(8-amino-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate (9H-fluoren-9-yl)methyl(8-acetamido-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate (2.0 g, 3.21 mmol, 80%) was dissolved in dioxane (20 mL), then 12 mol / L concentrated hydrochloric acid (5 mL) was added, and the temperature was raised to 70°C. The reaction was carried out for 2 hours and monitored by LC-MS. The reaction solution was poured into water (40 mL), then extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated brine (40 mL), dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure to obtain the crude product. The crude product was purified by column chromatography using a silica gel column (PE:EA = 2:1) to obtain 740 mg of the title compound.

[0618] The structural characteristics data was as follows: ESI-MS (m / z): 457.3[M+H] + .

[0619] Step 6: Synthesis of (9H-fluoren-9-yl)methyl((9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)carbamate (S)-4-ethyl-4-hydroxyl-7,8-dihydro-1H-pyrano[3,4-f]indolidine-3,6,10(4H)-trione (442 mg, 1.68 mmol) and (9H-fluoren-9-yl)methyl(8-amino-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate (640 mg, 1.40 mmol) were added to toluene (30 mL), followed by the addition of p-toluenesulfonic acid (242 mg, 1.40 mmol). After the addition, the temperature was raised to 135°C and the reaction was carried out for 2 hours. The reaction solution was directly evaporated to dryness under reduced pressure at 140°C to obtain the crude product. The crude product was purified by column chromatography using a silica gel column (DCM:MeOH=33:1) to obtain 1.02 g of the title compound.

[0620] The structural characteristics data was as follows: ESI-MS (m / z): 684.1[M+H] + .

[0621] Step 7: Synthesis of (1S,9S)-1-amino-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione (compound 5-29-1) and (1R,9S)-1-amino-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione (9H-fluoren-9-yl)methyl((9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4:6,7]indolidino[1,2-b]quinoline-1-yl) carbamate (1.02 g, 1.49 mmol) is dissolved in N,N-dimethylformamide (15 mL). The mixture was dissolved, then 5 ml of diethylamine was added. After addition, the reaction was carried out at room temperature for 0.5 hours, monitored by LC-MS, and the reaction solution was distilled under reduced pressure to remove ethylenediamine. The pH was then adjusted to 2-3 with 1 mol / L hydrochloric acid aqueous solution, and the reaction solution was then directly purified by preparative high-performance liquid chromatography to obtain two isomers of the title compound (5-22-7-A: 60 mg; 5-22-7-B: 55 mg).

[0622] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 35]

[0623] The structural characteristics data for 5-22-7-A (6-minute LCMS, with an earlier peak and retention time of 2.28 minutes) were as follows: ESI-MS (m / z): 462.2[M+H] + . The structural characteristics data for 5-22-7-B (6-minute LCMS, with a later peak having a retention time of 2.35 minutes) were as follows: ESI-MS (m / z): 462.2[M+H] + .

[0624] Step 8: Synthesis of N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-1,2,3,9,10,12,13,15-tetrahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide and N-((1R,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-1,2,3,9,10,12,13,15-tetrahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide At 25°C, a single stereoisomer compound 5-28-7-A (40 mg, 86 μmol) and glycolic acid (8 mg, 104 μmol) were dissolved in DMF (2 mL). Then, HATU (40 mg, 104 μmol) and DIPEA (36 mg, 258 μmol) were added, and the reaction was carried out at room temperature for 0.5 hours. The reaction was monitored by LC-MS, and the reaction solution was directly purified by preparative high-performance liquid chromatography to obtain compound 5-22-A (12.5 mg).

[0625] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 36]

[0626] The structural characteristics data was as follows: The structural characteristics data for 5-22-A (6-minute LCMS, with an earlier peak and retention time of 2.540 minutes) were as follows: ESI-MS (m / z): 520.0[M+H] + . 1 HNMR (400 MHz, DMSO-d6) δ 8.46 (d, J = 8.9Hz, 1H), 7.74 (d, J = 11.9 Hz, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.64 - 5.56 (m,1H), 5.49 (t, J = 5.8 Hz, 1H), 5.42 (s, 2H), 5.19 (s, 2H), 3.96 (d, J = 5.7 Hz,2H), 2.25 - 2.10 (m, 2H), 2.04 - 1.79 (m, 4H), 1.23 (s, 2H), 1.15 - 1.05 (m,2H), 0.87 (t, J = 7.2 Hz, 3H), 0.80 - 0.70 (m, 2H).

[0627] At 25°C, a single stereoisomer compound 5-28-7-B (30 mg, 65 μmol) and glycolic acid (6 mg, 78 μmol) were dissolved in DMF (2 mL). Then, HATU (40 mg, 104 μmol) and DIPEA (17 mg, 130 μmol) were added, and the reaction was carried out at room temperature for 0.5 hours. The reaction was monitored by LC-MS, the reaction solution was concentrated to dryness, and the compound 5-22-B (13.83 mg) was directly purified by preparative high-performance liquid chromatography.

[0628] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 37]

[0629] The structural characteristics data was as follows: The structural characteristics data for 5-22-B (6-minute LCMS, with an earlier peak and retention time of 2.612 minutes) were as follows: ESI-MS (m / z): 520.0[M+H] + . 1 HNMR (400 MHz, DMSO-d6) δ 8.49 (d, J = 8.9Hz, 1H), 7.74 (d, J = 11.9 Hz, 1H), 7.31 (s, 1H), 6.53 (s, 1H), 5.60 (s, 1H),5.51 (t, J = 5.9 Hz, 1H), 5.43 (s, 2H), 5.25 - 5.13 (m, 2H), 3.97 (d, J = 5.8Hz, 2H), 2.18 (s, 2H), 2.04 - 1.91 (m, 4H), 1.90 - 1.80 (m, 1H), 1.23 (s, 6H),1.15 - 1.05(m, 2H), 0.87 (t, J = 7.2 Hz, 4H), 0.80 - 0.70 (m, 2H).

[0630] Example 20: Synthesis of (R)-3-(dimethylamino)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropionamide and (S)-3-(dimethylamino)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropionamide [ka]

[0631] Step 1: (9H-fluoro-9-yl)methyl((S)-3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2-hydroxy-3-oxopropyl)carbame Synthesis of (9H-fluoro-9-yl)methyl((R)-3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2-hydroxy-3-oxopropyl)carbamate At 25℃, (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-2,3,12,15-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13(1H,9H)-dione mesylate (72 mg, 166 μmol) and 3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-hydroxypropionic acid (65 mg (g, 199 μmol) was dissolved in DMF (2 mL), then HATU (95 mg, 250 μmol) and DIPEA (65 mg, 498 μmol) were added, and the reaction was carried out at room temperature for 0.5 hours. The reaction was monitored by LC-MS, the reaction solution was concentrated to dryness, and the solution was directly purified by preparative high-performance liquid chromatography to obtain the title compounds (compound 1-10-1-A, 24 mg; and compound 1-10-1-B, 28 mg).

[0632] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 38]

[0633] The structural characteristics data for compound 1-10-1-A (6 min LCMS, earlier peak with retention time of 3.283 min) were as follows: ESI-MS (m / z): 745.4[M+H] + . The structural characteristics data for compound 1-10-1-B (later peak with retention time of 3.465 minutes, measured by 6 min LCMS) were as follows: ESI-MS (m / z): 745.4[M+H] + .

[0634] Step 2: Synthesis of (R)-3-amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropionamide and (S)-3-amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropionamide At 25°C, compound 1-10-1-B (28 mg, 37 μmol) was dissolved in DMF (2 mL), then diethylamine (1 mL) was added, and the reaction was carried out at room temperature for 1.0 hour. The reaction solution was concentrated to dryness to obtain 28 mg of crude product (compound 1-10-2-B), which was used directly in the next reaction.

[0635] At 25°C, compound 1-10-1-A (24 mg, 33 μmol) was dissolved in DMF (2 mL), then diethylamine (1 mL) was added, and the reaction was carried out at room temperature for 1.0 hour. The reaction solution was concentrated to dryness to obtain 24 mg of crude product (compound 1-10-2-A), which was used directly in the next reaction.

[0636] The structural characteristics data was as follows: ESI-MS (m / z): 523.2[M+H] + .

[0637] Step 3: Synthesis of (R)-3-(dimethylamino)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropionamide and (S)-3-(dimethylamino)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropionamide At 25°C, compound 1-10-2-B (28 mg, 37 μmol, 70%) was dissolved in methanol (2 mL), then formaldehyde solution (1 mL) was added, and the reaction was carried out at room temperature for 16.0 hours. Then sodium borohydride cyanohydride (7.07 mg, 96.45 μmol) was added, and the reaction was carried out at room temperature for 1.0 hour. The reaction was monitored by LC-MS, the reaction solution was concentrated to dryness, and the solution was directly purified by preparative high-performance liquid chromatography to obtain 1.3 mg of the title compound (compound 1-10B).

[0638] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: methanol; Mobile phase B: water (0.05% formic acid) [Table 39]

[0639] The structural characteristics data for compound 1-10-B (6 min LCMS, earlier peak with retention time of 1.937 min) were as follows: ESI-MS (m / z): 551.2[M+H] + . 1HNMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 8.6Hz, 1H), 8.31 (s, 2H), 7.80 (d, J = 11.0 Hz, 1H), 7.31 (s, 1H), 6.55 (s, 1H),5.54 (s, 1H), 5.43 (s, 2H), 5.34 (d, J = 19.2 Hz, 1H), 5.19 (d, J = 19.1 Hz,1H), 4.09 - 4.06 (m, 1H), 3.20 - 3.15 (m, 2H), 2.59 - 2.53 (m, 1H), 2.45 -2.42(m, 1H), 2.42 - 2.38 (s, 3H), 2.23 - 2.19 (d, J = 7.0 Hz, 1H), 2.13 (s, 6H),2.12 - 2.08 (m, 1H), 2.02 - 1.95 (m, 1H), 1.90 - 1.85 (m, 2H), 1.23 (s, 2H),0.88 (d, J = 7.2 Hz, 3H).

[0640] At 25°C, compound 1-10-2-A (24 mg, 33 μmol, 70%) was dissolved in methanol (2 mL), then formaldehyde solution (1 mL) was added, and the reaction was carried out at room temperature for 16.0 hours. Then sodium borohydride cyanohydride (6.06 mg, 96.45 μmol) was added, and the reaction was carried out at room temperature for 1.0 hour. The reaction was monitored by LC-MS, the reaction solution was concentrated to dryness, and the solution was directly purified by preparative high-performance liquid chromatography to obtain 4.44 mg of the title compound (compound 1-10-A).

[0641] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: methanol; Mobile phase B: water (0.05% formic acid) [Table 40]

[0642] The structural characteristics data for compound 1-10-A (6 min LC-MS, earlier peak with retention time of 1.920 min) were as follows: ESI-MS (m / z): 551.2[M+H] + . 1 HNMR (400 MHz, DMSO-d6) δ 8.58 (d, J = 9.0Hz, 1H), 8.28 (s, 1H), 7.84 (d, J = 10.9 Hz, 1H), 7.37 (s, 1H), 6.61 (s, 1H),5.66 - 5.59 (m, 1H), 5.49 (s, 2H), 5.35 (d, J = 19.1 Hz, 1H), 5.17 (d, J = 18.9Hz, 1H), 4.23 - 4.16 (m, 1H), 3.23 (d, J = 7.8 Hz, 2H), 2.75 - 2.67 (m, 2H),2.45 (s, 3H), 2.30 (s, 6H), 2.27 - 2.17 (m, 2H), 2.14 - 1.99 (m, 1H), 1.98 -1.87 (m, 2H), 1.30 (s, 2H), 0.93 (t, J = 7.3 Hz, 3H).

[0643] Example 21: (S)-14-(2-(cyclopropylamino)ethyl)-7-ethyl-7-hydroxy-7H-[1,3]dioxolano[4,5-g]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-8,11(10H,13H)-dione (compound 4-14) [ka]

[0644] Step 1: Synthesis of 2-nitro-4,5-methylenedioxyacetophenone Compound 4-14-1 (10.0 g, 60.92 mmol) was dissolved in nitromethane (100 mL), and concentrated nitric acid (26 mL) was slowly added dropwise while stirring. The reaction was allowed to proceed at room temperature for 2 hours. Monitoring the reaction by TLC showed that a small amount of the starting material remained, and the product was clearly defined. The reaction solution was neutralized by slowly adding saturated sodium bicarbonate aqueous solution dropwise, and extraction was performed three times by adding dichloromethane. The organic phases were combined, washed three times with saturated brine, dried, and concentrated to obtain the crude product. This was purified by silica gel column chromatography (eluent: 0-20% ethyl acetate / petroleum ether) to obtain 9.8 g of the title compound.

[0645] Step 2: Synthesis of 6-amino-3,4-methylenedioxyacetophenone Compound 4-14-2 (2.0 g, 9.56 mmol) was dissolved in ethyl acetate (20 mL), then carbon-supported 10% palladium (0.2 g) was added, and the mixture was subjected to hydrogen displacement and reacted for 4 hours with stirring under hydrogen protection. After filtration, the filtrate was concentrated under reduced pressure to obtain 1.7 g of the crude product of the title compound.

[0646] Step 3: Synthesis of 6-acetamido-3,4-methylenedioxyacetophenone Compound 4-14-3 (1.7 g, 9.49 mmol) was dissolved in acetic anhydride (17 mL) and reacted with stirring for 1 hour. After evaporating the solvent under reduced pressure, water was added, the mixture was stirred, the solid was filtered, washed with water, and dried under vacuum to obtain 2.08 g of the crude product of the title compound.

[0647] Step 4: Synthesis of (E)-N-(6-(3-(dimethylamino)acryloyl)benzo[d][1,3]dioxol-5-yl)acetamide Compound 4-14-4 (1.88 g, 8.50 mmol) was dissolved in DMF-DMA (30 mL), heated to 120°C, and reacted for 2 hours. The solvent was evaporated under reduced pressure to obtain 2.33 g of the crude product of the title compound.

[0648] The structural characteristics data was as follows: ESI-MS (m / z): 277.2[M+1]+.

[0649] Step 5: Synthesis of (E)-N-(6-(3-(cyclopropylamino)acryloyl)benzo[d][1,3]dioxol-5-yl)acetamide Compound 4-14-5 (200 mg, 0.72 mmol) was dissolved in ethanol (5 mL), and then cyclopropylamine (413.3 mg, 7.24 mmol) was added dropwise. The mixture was heated to 50°C and reacted for 16 hours. The solvent was evaporated under reduced pressure to obtain 208 mg of the crude product of the title compound.

[0650] The structural characteristics data was as follows: ESI-MS (m / z): 289.2[M+1]+.

[0651] Step 6: Synthesis of N-(6-(3-(cyclopropylamino)propionyl)benzo[d][1,3]dioxol-5-yl)acetamide Compound 4-14-6 (208 mg, 0.72 mmol) was dissolved in glacial acetic acid (4 mL), and sodium borohydride (13.65 mg, 0.36 mmol) was added while stirring in an ice bath. The mixture was then heated to room temperature and reacted with stirring for 3 hours. The solvent was evaporated under reduced pressure to obtain 209 mg of the crude product of the title compound.

[0652] The structural characteristics data was as follows: ESI-MS (m / z): 291.1[M+1]+.

[0653] Step 7: Synthesis of (9H-fluoren-9-yl)methyl(3-(6-acetamidobenzo[d][1,3]dioxol-5-yl)-3-oxopropyl)(cyclopropyl)carbamate Compound 4-14-7 (200 mg, 0.69 mmol) was dissolved in 1,4-dioxane (20 mL) and water (20 mL). Then, while stirring, 9-fluorenylmethyl-N-succinimide carbonate (395 mg, 0.68 mmol) and sodium bicarbonate (231.5 mg, 2.76 mmol) were added, and the mixture was reacted at room temperature for 2 hours. Water and ethyl acetate were added while stirring, and the mixture was allowed to stand for liquid separation. The organic phase was washed with saturated brine, dried, concentrated, and purified by silica gel column chromatography (eluent: 30% ethyl acetate / petroleum ether) to obtain 350 mg of the title compound.

[0654] Step 8: Synthesis of (9H-fluoren-9-yl)methyl(3-(6-aminobenzo[d][1,3]dioxol-5-yl)-3-oxopropyl)(cyclopropyl)carbamate Compound 4-14-8 (350 mg, 0.68 mmol) was dissolved in 1,4-dioxane (10 mL), then 3N hydrochloric acid aqueous solution (10 mL) was added dropwise, and the mixture was heated to 60°C and reacted with stirring for 16 hours. Water and ethyl acetate were added, the mixture was stirred, and it was allowed to stand for liquid separation. The organic phase was washed with water, dried, concentrated, and purified by silica gel column chromatography (eluent: 33% ethyl acetate / petroleum ether) to obtain 218 mg of the title compound.

[0655] Step 9: Synthesis of (S)-(9H-fluoren-9-yl)methylcyclopropyl(2-(7-ethyl-7-hydroxy-8,11-dioxo-8,10,11,13-tetrahydro-7H-[1,3]dioxolano[4,5-g]pyrano[3,4]:6,7]indolidino[1,2-b]quinoline-14-yl)ethyl)carbamate Compound 4-14-9 (40 mg, 0.085 mmol) and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolidine-3,6,10(4H)-trione (24.62 mg, 0.094 mmol) were dissolved in toluene (1 mL), then p-toluenesulfonic acid (2.93 mg, 0.017 mmol) was added, and the mixture was heated to 120°C and reacted for 4 hours. After concentration under reduced pressure, 59 mg of the crude product of the title compound was obtained.

[0656] The structural characteristics data was as follows: ESI-MS (m / z): 698.1[M+1]+.

[0657] Step 10: Synthesis of (S)-14-(2-(cyclopropylamino)ethyl)-7-ethyl-7-hydroxy-7H-[1,3]dioxolano[4,5-g]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-8,11(10H,13H)-dione

[0658] Compound 4-14-10 (59 mg, 0.085 mmol) was dissolved in DMF (1 mL), and then diethylamine (0.5 mL) was added dropwise. The mixture was reacted for 1 hour with stirring. The reaction solution was distilled under reduced pressure to remove the diethylamine, and the mixture was acidified by adding 3N hydrochloric acid dropwise. The solution was purified by HPLC (purification conditions were as follows), and lyophilized to obtain 12.66 mg of the trifluoroacetate salt of the title compound.

[0659] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% trifluoroacetic acid) [Table 41]

[0660] The structural characteristics data was as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.73 (s, 2H), 7.67(s, 1H), 7.57 (s, 1H), 7.26 (s, 1H), 6.54 (s, 1H), 6.33 (s, 2H), 5.44 (s, 2H),5.34 (s, 2H), 3.40 (s, 4H), 2.82 (s, 1H), 1.91 - 1.81 (m, 2H), 0.87 (t, J = 7.2Hz, 5H), 0.79 (d, J = 7.4 Hz, 2H). ESI-MS (m / z): 476.1[M+1]+.

[0661] Example 22: (S)-7-ethyl-7-hydroxy-14-(2-((2-methoxyethyl)amino)ethyl)-7H-[1,3]dioxolano[4,5-g]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-8,11(10H,13H)-dione (compound 4-15) [ka]

[0662] Step 1: Synthesis of (E)-N-(6-(3-((2-methoxyethyl)amino)acryloyl)benzo[d][1,3]dioxol-5-yl)acetamide Compound 4-14-5 (200 mg, 0.72 mmol) was dissolved in ethanol (5 mL), and then 2-methoxyethylamine (543.7 mg, 7.24 mmol) was added dropwise. The mixture was heated to 50°C and reacted for 16 hours. Monitoring the reaction by LC-MS showed that a small amount of starting material remained, and the product was clearly defined. The solvent was evaporated under reduced pressure to obtain 221 mg of the title compound, which was used directly in the next reaction.

[0663] The structural characteristics data was as follows: ESI-MS (m / z):307.1[M+1]+

[0664] Step 2: Synthesis of N-(6-(3-((2-methoxyethyl)amino)propionyl)benzo[d][1,3]dioxol-5-yl)acetamide Compound 4-15-1 (200 mg, 0.65 mmol) was dissolved in glacial acetic acid (4 mL), and then sodium borohydride (12.35 mg, 0.33 mmol) was added while stirring in an ice bath. The mixture was heated to room temperature and reacted with stirring for 3 hours. Monitoring the reaction by LC-MS showed that the starting material disappeared and the product was clearly defined. The solvent was evaporated under reduced pressure to obtain 200 mg of the title compound, which was then used directly in the next reaction.

[0665] ESI-MS (m / z):309.1[M+1]+

[0666] Step 3: Synthesis of (9H-fluoren-9-yl)methyl(3-(6-acetamidobenzo[d][1,3]dioxol-5-yl)-3-oxopropyl)(2-methoxyethyl)carbamate Crude product of compound 4-15-2 (200 mg, 0.65 mmol) was dissolved in 1,4-dioxane (20 mL) and water (20 mL). Then, while stirring, 9-fluorenylmethyl-N-succiniliminocarbonate (372 mg, 0.65 mmol) and sodium bicarbonate (231.5 mg, 2.76 mmol) were added, and the mixture was reacted at room temperature for 2 hours. Monitoring the reaction by TLC showed that the starting materials had disappeared and the product was clearly visible. Water and ethyl acetate were added, the mixture was stirred, and allowed to stand for liquid separation. The organic phase was washed with saturated brine, dried, concentrated, and purified by silica gel column (eluent: 50% ethyl acetate / petroleum ether) to obtain 180 mg of the title compound.

[0667] Step 4: Synthesis of (9H-fluoren-9-yl)methyl(3-(6-aminobenzo[d][1,3]dioxol-5-yl)-3-oxopropyl)(2-methoxyethyl)carbamate Compound 4-15-3 (180 mg, 0.68 mmol) was dissolved in 1,4-dioxane (5 mL), and 3N hydrochloric acid aqueous solution (5 mL) was added dropwise. The mixture was heated to 60°C and reacted with stirring for 16 hours. Water and ethyl acetate were added, the mixture was stirred, and the mixture was allowed to stand for liquid separation. The organic phase was washed with water, dried, concentrated, and purified by silica gel column chromatography (eluent: 45% ethyl acetate / petroleum ether) to obtain 132 mg of the title compound.

[0668] Step 5: Synthesis of (S)-(9H-fluoren-9-yl)methyl(2-(7-ethyl-7-hydroxy-8,11-dioxo-8,10,11,13-tetrahydro-7H-[1,3]dioxolano[4,5-g]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-14-yl)ethyl)(2-methoxyethyl)carbamate Compound 4-15-4 (130 mg, 0.266 mmol) and rac-(4S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolidine-3,6,10-trione (70.05 mg, 0.266 mmol) were dissolved in toluene (4 mL), and then p-toluenesulfonic acid (9.16 mg, 0.053 mmol) was added. The mixture was heated to 120°C and reacted for 4 hours. Monitoring the reaction by LC-MS showed that the starting materials disappeared and the product was clearly defined. After concentration under reduced pressure, 190 mg of the crude product of the title compound was obtained.

[0669] The structural characteristics data was as follows: ESI-MS (m / z): 716.1[M+1]+

[0670] Step 6: Synthesis of (S)-7-ethyl-7-hydroxy-14-(2-((2-methoxyethyl)amino)ethyl)-7H-[1,3]dioxolano[4,5-g]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-8,11(10H,13H)-dione Compound 4-15-5 (190 mg, 0.265 mmol) was dissolved in DMF (3 mL), and then diethylamine (2 mL) was added dropwise. The reaction was allowed to proceed for 1 hour with stirring. Monitoring the reaction by LC-MS showed that the starting material disappeared and the product was clearly defined. The reaction solution was distilled under reduced pressure to remove the diethylamine, acidified by adding 3N hydrochloric acid dropwise, purified, and freeze-dried to obtain 99.28 mg of the title compound.

[0671] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 42]

[0672] The structural characteristics data was as follows: 1 HNMR (400 MHz, DMSO-d6): δ 8.68 (s, 2H), 7.68(s, 1H), 7.56 (s, 1H), 7.26 (s, 1H), 6.52 (s, 1H), 6.32 (s, 2H), 5.44 (s, 2H),5.31 (s, 2H), 3.65 - 3.58 (m, 2H), 3.42 (d, J = 10.2 Hz, 2H), 3.36 (s, 3H),3.22 (d, J = 4.0 Hz, 4H), 1.94 - 1.80 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H). ESI-MS (m / z):494.2[M+1]+

[0673] Example 23: N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide and N-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide [ka]

[0674] Step 1: Synthesis of N-(3-bromo-5-fluoro-4-methoxyphenyl)acetamide 3-Bromo-5-fluoro-4-methoxyaniline (1.7 g, 7.73 mmol) was dissolved in tetrahydrofuran (30 mL), then triethylamine (2.35 g, 23.18 mmol) and acetic anhydride (1.18 g, 11.59 mmol) were added. After the addition, the temperature was raised to 50°C, the reaction mixture was stirred for 4 hours, and monitored by LC-MS. After the reaction solution was cooled to room temperature, it was diluted with ethyl acetate (50 mL), then washed once each with water (30 mL) and saturated brine (30 mL) to separate the organic phase. The organic phase was dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. This was then slurryed (using petroleum ether:ethyl acetate = 5:1) and purified to obtain 1.1 g of the title compound.

[0675] The structural characteristics data was as follows: ESI-MS (m / z): 262.0[M+H] + .

[0676] Step 2: Synthesis of (E)-4-(5-acetylamino-3-fluoro-2-methoxyphenyl)-3-butenic acid N-(3-bromo-5-fluoro-4-methoxyphenyl)acetamide (1.1 g, 4.20 mmol) and 3-butenoic acid (397.47 mg, 4.62 mmol) were dissolved in 1,4-dioxahexane (20 mL) and water (5 mL). Then triethylamine (1.27 g, 12.59 mmol), tris(o-methylphenyl)phosphine (127.75 mg, 419.73 μmol), and palladium acetate (47.12 mg, 209.89 μmol) were added. After the addition, the reaction system was subjected to nitrogen purging three times, heated to 100 °C, and reacted under a nitrogen atmosphere for 4 hours. The reaction was monitored by LC-MS, and after the reaction solution was cooled to room temperature, 1 mol / L aqueous sodium hydroxide solution (50 mL) and ethyl acetate (50 mL) were added, and the mixture was shaken for liquid separation. The aqueous phase was separated, the pH was adjusted to approximately 3 with a 4 mol / L hydrochloric acid aqueous solution, then extracted with ethyl acetate (40 mL x 2), the organic phase was combined, washed with saturated brine (40 mL), dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain 1.1 g of the crude product of the title compound.

[0677] The structural characteristics data was as follows: ESI-MS (m / z): 268.1[M+H] + .

[0678] Step 3: Synthesis of 4-(5-acetylamino-3-fluoro-2-methoxyphenyl)butanoic acid Crude product (1.1 g, 4.12 mmol) of (E)-4-(5-acetylamino-3-fluoro-2-methoxyphenyl)-3-butenoic acid was dissolved in methanol (20 mL), then carbon-supported 10% palladium (100 mg) was added. After addition, the reaction system was subjected to hydrogen displacement three times using a hydrogen balloon, and the reaction was carried out under a hydrogen atmosphere for 4 hours. The reaction solution was monitored by LC-MS, the reaction solution was filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain 1.05 g of the crude product of the title compound.

[0679] The structural characteristics data was as follows: ESI-MS (m / z): 270.1[M+H] + .

[0680] Step 4: Synthesis of N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide Crude product of 4-(5-acetylamino-3-fluoro-2-methoxyphenyl)butanoic acid (1.1 g, 4.09 mmol) was dissolved in trifluoroacetic acid (10 mL), cooled to 5°C, and then trifluoroacetic anhydride (4.29 g, 20.43 mmol) was slowly added. After the addition, the reaction system was allowed to rise naturally to room temperature and reacted for 2 hours. The reaction was monitored by LC-MS, and the reaction solution was slowly poured into water (60 mL). Then it was extracted with ethyl acetate (40 mL x 3), the organic phases were combined, washed with saturated sodium bicarbonate aqueous solution until neutral, then washed with saturated brine (40 mL), dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product. This was purified by flash silica gel column (ethyl acetate:petroleum ether = 0-40%) to obtain 503 mg of the title compound.

[0681] The structural characteristics data was as follows: ESI-MS (m / z): 252.1[M+H] + .

[0682] Step 5: Synthesis of N-(3-fluoro-7-(hydroxyimino)-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide Add tetrahydrofuran (15 mL) and tert-butanol (4 mL) to the reaction flask, cool to 5°C in an ice bath, add potassium tert-butoxide (491.26 mg, 4.38 mmol), then dissolve N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (500 mg, 1.99 mmol) in tetrahydrofuran (5 mL) and slowly add it dropwise. Subsequently, 10 minutes later, isoamyl nitrite (373.01 mg, 3.18 mmol) was added, and the reaction was carried out at 5°C for 1 hour, monitored by LC-MS, the reaction solution was quenched with saturated ammonium chloride aqueous solution (50 mL), extracted with ethyl acetate (40 × 2), the organic phases were combined, washed with saturated brine (40 mL), dehydrated with anhydrous sodium sulfate, then filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain 550 mg of the crude product of the title compound.

[0683] The structural characteristics data was as follows: ESI-MS (m / z): 281.1[M+H] + .

[0684] Step 6: Synthesis of N-(7-amino-3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide Crude product (520 mg, 1.86 mmol) of N-(3-fluoro-7-(hydroxyimino)-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide was dissolved in a mixed solution of methanol (10 mL) and tetrahydrofuran (10 mL). Then, 3.71 mL of 1 mol / L aqueous hydrochloric acid solution and 50 mg of carbon-supported 10% palladium were added. After addition, the reaction system was subjected to hydrogen displacement three times using a hydrogen balloon. The reaction was carried out at room temperature under a hydrogen atmosphere for 1 hour, monitored by LC-MS, filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain 551 mg of the crude hydrochloride salt of the title compound.

[0685] The structural characteristics data was as follows: ESI-MS (m / z): 267.1[M+H] + .

[0686] Step 7: Synthesis of (9H-fluoren-9-yl)methyl(8-acetamido-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate Crude hydrochloride of N-(7-amino-3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide (550 mg, 1.64 mmol) was dissolved in 1,4-dioxane (15 mL), then sodium bicarbonate (549.45 mg, 6.54 mmol), water (5 mL), and 9-fluorenylmethyl-N-succinimide carbonate (1.12 g, 1.96 mmol) were added. The reaction was carried out at room temperature for 2 hours with stirring, monitored by LC-MS, and the reaction solution was poured into water (50 mL). The solution was then extracted with ethyl acetate (40 mL x 2), the organic phases were combined, washed with saturated brine (40 mL), dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure to obtain the crude product. The crude product was purified using a C18 (acetonitrile / 0.05% formic acid aqueous solution, 20% to 100% acetonitrile) reverse-phase column to obtain 410 mg of the title compound.

[0687] The structural characteristics data was as follows: ESI-MS (m / z): 489.1[M+H] + .

[0688] Step 8: Synthesis of (9H-fluoren-9-yl)methyl(8-amino-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate (9H-fluoren-9-yl)methyl(8-acetamido-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate (410 mg, 839.29 μmol) was dissolved in dioxane (10 mL), 12 mol / L concentrated hydrochloric acid (2 mL) was added, and the temperature was raised to 70°C. The reaction was carried out for 2 hours and monitored by LC-MS. The reaction solution was poured into water (30 mL), then extracted with ethyl acetate (30 mL x 2), the organic phases were combined, washed with saturated brine (30 mL), dehydrated with anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure to obtain the crude product. This was purified using flash silica gel (ethyl acetate: petroleum ether = 0-60%) to obtain 351 mg of the title compound.

[0689] The structural characteristics data was as follows: ESI-MS (m / z): 447.1[M+H] + .

[0690] Step 9: Synthesis of (9H-fluoren-9-yl)methyl((9S)-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)carbamate (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indolidine-3,6,10(4H)-trione (247.64 mg, 940.71 μmol) and (9H-fluoren-9-yl)methyl(8-amino-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate (350 mg, 783.93 μmol) were added to toluene (15 mL), followed by the addition of p-toluenesulfonic acid (134.84 mg, 783.93 μmol). After the addition, the temperature was raised to 135°C and the reaction was carried out for 4 hours. The reaction solution was directly evaporated to dryness under reduced pressure at 135°C to obtain the crude product, which was purified by flash silica gel (methanol:dichloromethane = 0-6%) column to obtain 358 mg of the title compound.

[0691] The structural characteristics data was as follows: ESI-MS (m / z): 674.2[M+H] + .

[0692] Step 10: Synthesis of (9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione (9H-fluoren-9-yl)methyl((9S)-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)carbamate (358 mg, 531.41 μmol) was dissolved in N,N-dimethylformamide (4 mL), then diethylamine (0.4 mL) was added, and the reaction was carried out at room temperature for 0.5 hours, monitored by LC-MS, and the reaction solution was evaporated to dryness under reduced pressure to obtain the crude product. This was then slurried in ethyl acetate to purify it and obtain 220 mg of the title compound.

[0693] The structural characteristics data was as follows: ESI-MS (m / z): 452.1[M+H] + .

[0694] Step 11: Synthesis of N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide and N-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide (9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione (50 mg, 110.76 μmol) and 2-hydroxyacetic acid (16.85 mg, 221.51 μmol) are mixed with N,N-dimethylformamide ( The solution was dissolved in 2 mL of the solution, then HATU (84.17 mg, 221.51 μmol) and N,N-diisopropylethylamine (42.94 mg, 332.27 μmol) were added. The reaction was then carried out at room temperature for 0.5 hours, monitored by LC-MS, and the reaction solution was directly purified by preparative high-performance liquid chromatography to obtain two isomers (5-34-A: 6.22 mg, 5-34-B: 9.81 mg).

[0695] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) [Table 43]

[0696] The structural characteristics data for 5-34-A were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.53 (d, J = 9.0Hz, 1H), 7.89 (d, J = 12.3 Hz, 1H), 7.33 (s, 1H), 5.58 (q, J = 7.5, 7.0 Hz,1H), 5.42 (d, J = 2.1 Hz, 2H), 5.25 - 5.09 (m, 2H), 3.98 (s, 2H), 3.96 (d, J =1.1 Hz, 3H), 3.30 - 3.10 (m, 2H), 2.15 (q, J = 7.4 Hz, 2H), 1.93 - 1.80 (m,2H), 0.87 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 510.2[M+H] + . The structural characteristics data for 5-34-B were as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.55 (d, J = 9.0Hz, 1H), 7.90 (d, J = 12.3 Hz, 1H), 7.34 (s, 1H), 5.59 (q, J = 7.4, 6.8 Hz,1H), 5.43 (s, 2H), 5.25 - 5.11 (m, 2H), 3.99 (s, 2H), 3.96 (d, J = 1.1 Hz, 3H),3.30 - 3.13 (m, 2H), 2.15 (q, J = 6.4 Hz, 2H), 1.93 - 1.82 (m, 2H), 0.88 (t, J= 7.3 Hz, 3H). ESI-MS (m / z): 510.2[M+H] + .

[0697] Example 24: (2R)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine and (2R)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine [ka]

[0698] Step 1: Synthesis of (2R)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine and (2R)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine (9S)-1-amino-5-chloro-9-ethyl-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione (50 mg, 110.64 μmol) and (2R)-2-hydroxypropanoic acid (19.93 mg, 221.29 μmol) are mixed with N,N-dimethylformamide ( The compound was dissolved in 2 mL of solution, then HATU (84.09 mg, 221.29 μmol) and N,N-diisopropylethylamine (42.90 mg, 331.93 μmol) were added. After addition, the reaction was carried out at room temperature for 0.5 hours, monitored by LC-MS, and the reaction solution was directly purified by preparative high-performance liquid chromatography to obtain two isomers of the title compound (2-27-A: 5.73 mg, 2-27-B: 7.59 mg).

[0699] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% trifluoroacetic acid) [Table 44]

[0700] The structural characteristics data for 2-27-A were as follows: 1HNMR (400 MHz, DMSO-d6) δ 8.45 (d, J = 9.1Hz, 1H), 8.14 (s, 1H), 7.30 (s, 1H), 6.54 (s, 1H), 5.63 (s, 1H), 5.56 (q, J =8.0 Hz, 1H), 5.42 (s, 2H), 5.25 (d, J = 19.0 Hz, 1H), 5.08 (d, J = 19.0 Hz,1H), 4.13 (q, J = 6.7 Hz, 1H), 3.27 - 3.12 (m, 2H), 2.51 (s, 3H), 2.23 - 2.13(m, 2H), 1.92 - 1.80 (m, 2H), 1.41 (d, J = 6.8 Hz, 3H), 0.87 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 524.2 [M+H] + . 2-27-B's structural characteristics are as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.40 (d, J = 8.9Hz, 1H), 8.15 (s, 1H), 7.31 (s, 1H), 6.54 (s, 1H), 5.55 - 5.49 (m, 1H), 5.43(d, J = 2.2 Hz, 2H), 5.18 (q, J = 19.0 Hz, 2H), 4.13 (q, J = 6.6 Hz, 1H), 3.24- 3.12 (m, 2H), 2.51 (s, 3H), 2.22 - 2.10 (m, 2H), 1.92 - 1.82 (m, 2H), 1.30(d, J = 6.7 Hz, 3H), 0.88 (t, J = 7.3 Hz, 3H). ESI-MS (m / z): 524.2 [M+H] + .

[0701] Example 25: Preparation of (R)-N-((1S,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine and (R)-N-((1R,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxypropylamine [ka]

[0702] At 25°C, 3-1-A (20.0 mg, 43.9 μmol) and D-lactic acid (7.90 mg, 87.8 μmol) were dissolved in DMF (1.0 mL), then HATU (33.4 mg, 87.8 μmol) and DIPEA (17.0 mg, 131.6 μmol) were added, and the mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated to remove most of the DMF, and the residue was purified by preparative high-performance liquid chromatography to obtain compound 3-26-A (15.4 mg, yield 64%).

[0703] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) Holding time: 5.3~6.2 minutes [Table 45]

[0704] The structural characteristics data was as follows: MS m / z (ESI): 528.2 [M+H] + 1HNMR (400 MHz, DMSO-d6) δ 8.52 (d, J = 9.2Hz, 1H), 8.05 (d, J = 10.0 Hz, 1H), 7.33 (s, 1H), 6.55 (s, 1H), 5.62 - 5.59 (m,2H), 5.43 (s, 2H), 5.28 - 5.10 (m, 2H), 4.13 - 4.11 (m, 1H), 3.41 - 3.38 (m,1H), 3.28 - 3.22 (m, 1H), 2.20 - 2.18 (m, 2H), 1.92 - 1.80 (m, 2H), 1.40 (d, J= 6.8 Hz, 3H), 0.87 (t, J = 7.2 Hz, 3H).

[0705] At 25°C, 3-1-B (20.0 mg, 43.9 μmol) and D-lactic acid (7.90 mg, 87.8 μmol) were dissolved in DMF (1.0 mL), then HATU (33.4 mg, 87.8 μmol) and DIPEA (17.0 mg, 131.6 μmol) were added, and the mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated to remove most of the DMF, and the residue was purified by preparative high-performance liquid chromatography to obtain compound 3-26-B (4.8 mg, yield 20%).

[0706] Chromatography column: SunFire Prep C18 OBD 19mm x 150mm x 5.0μm Mobile phase A: Acetonitrile; Mobile phase B: Water (0.05% formic acid) Holding time: 7.5~8.5 minutes [Table 46]

[0707] The structural characteristics data was as follows: MS m / z (ESI): 528.2 [M+H] + 1HNMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 8.8Hz, 1H), 8.06 (d, J = 10.4 Hz, 1H), 7.34 (s, 1H), 6.57 (br, 1H), 5.61 - 5.55(m, 1H), 5.48 - 5.39 (m, 2H), 5.27 - 5.16 (m, 2H), 4.15 - 4.10 (m, 1H), 3.32 -3.21 (m, 3H), 2.23 - 2.16 (m, 2H), 1.92 - 1.81 (m, 2H), 1.30 (d, J = 6.4 Hz,3H), 0.87 (t, J = 7.2 Hz, 3H).

[0708] Biological evaluation 1. Tumor cell proliferation inhibition test 1. Inhibitory effect of compounds on HT29 cell proliferation (1) Cell Plating: First, tumor cells HT29 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 1. [Table 47]

[0709] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0710] Detection of in vitro cell viability: After incubation, 50 μL of Cell Counting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (Cell Counting-Lite®), and vehicle RLU was obtained using cell-containing medium (Cell Counting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum inhibitory concentration (IC50) of the compound at half volume was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 2.

[0711] (2) Data results [Table 48]

[0712] The detection results showed that the compounds of the present invention shown in Table 2 had a potent inhibitory effect on the proliferation of HT29 colorectal cancer cells.

[0713] 2. Inhibitory effect of compounds on A549 cell proliferation (1) Cell Plating: First, tumor cells A549 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 3. [Table 49]

[0714] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0715] In vitro cell activity detection: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum half-volume inhibitory concentration (IC50) of the compound was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 4.

[0716] (2) Data results [Table 50]

[0717] The detection results showed that the compounds of the present invention shown in Table 4 had a significant inhibitory effect on the proliferation of A549 human lung cancer cells.

[0718] 3. Inhibitory effect of compounds on NCI-H1806 cell proliferation (1) Cell Plating: First, tumor cells HCC1806 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 5. [Table 51]

[0719] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0720] In vitro cell viability detection: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum inhibitory concentration (IC50) of the compound at half volume was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 6.

[0721] (2) Data results [Table 52]

[0722] The detection results showed that the compounds of the present invention shown in Table 6 had a significant inhibitory effect on the proliferation of HCC1806 human breast squamous cell carcinoma cells.

[0723] 4. Inhibitory effect of compounds on SKOV-3 cell proliferation (1) Cell Plating: First, tumor cells SKOV-3 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 7. [Table 53]

[0724] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0725] Detection of in vitro cell viability: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum inhibitory concentration (IC50) of the compound at half volume was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 8.

[0726] (2) Data results [Table 54]

[0727] The detection results showed that the compounds of the present invention shown in Table 8 had a significant inhibitory effect on the proliferation of SKOV-3 human ovarian cancer cells.

[0728] 5. Inhibitory effect of compounds on NCI-H358 cell proliferation (1) Cell Plating: First, tumor cells NCI-H358 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 9. [Table 55]

[0729] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0730] In vitro cell viability detection: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum inhibitory concentration (IC50) of the compound at half volume was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 10.

[0731] (2) Data results [Table 56]

[0732] The detection results showed that the compounds of the present invention shown in Table 10 had a significant inhibitory effect on the proliferation of NCI-H358 human non-small cell lung cancer cells.

[0733] 6. Inhibitory effects of compounds on NCI-N87 cell proliferation (1) Cell Plating: First, tumor cells NCI-N87 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 11. [Table 57]

[0734] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0735] In vitro cell viability detection: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum half-volume inhibitory concentration (IC50) of the compound was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 12.

[0736] (2) Data results [Table 58]

[0737] The detection results showed that the compounds of the present invention shown in Table 12 had a significant inhibitory effect on the proliferation of NCI-N87 human gastric cancer cells.

[0738] 7. Inhibitory effect of compounds on HELA cell proliferation (1) Cell Plating: First, tumor cells (Hela) were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 13. [Table 59]

[0739] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0740] In vitro cell viability detection: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum inhibitory concentration (IC50) of the compound at half volume was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 14.

[0741] (2) Data results [Table 60]

[0742] The detection results showed that the compounds of the present invention shown in Table 14 had a significant inhibitory effect on the proliferation of Hela human cervical cancer cells.

[0743] 8. Inhibitory effect of compounds on HCC70 cell proliferation (1) Cell Plating: First, tumor cells HCC70 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 15. [Table 61]

[0744] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0745] In vitro cell viability detection: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum inhibitory concentration (IC50) of the compound at half volume was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 16.

[0746] (2) Data results [Table 62]

[0747] The detection results showed that the compounds of the present invention shown in Table 16 had a significant inhibitory effect on the proliferation of HCC70 human breast cancer cells.

[0748] 9. Inhibitory effect of compounds on MDA-MB-231 cell proliferation (1) Cell Plating: First, tumor cells MDA-MB-231 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 17. [Table 63]

[0749] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0750] Detection of in vitro cell viability: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum inhibitory concentration (IC50) of the compound at half volume was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 18.

[0751] (2) Data results [Table 64]

[0752] The detection results showed that the compounds of the present invention shown in Table 18 had a significant inhibitory effect on the proliferation of MDA-MB-231 human breast cancer cells.

[0753] 10. Inhibitory effect of compounds on Jeko-1 cell proliferation (1) Cell Plating: First, tumor cells Jeko-1 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 19. [Table 65]

[0754] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0755] In vitro cell viability detection: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum inhibitory concentration (IC50) of the compound at half volume was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 20.

[0756] (2) Data results [Table 66]

[0757] The detection results showed that the compounds of the present invention shown in Table 20 had a significant inhibitory effect on the proliferation of Jeko-1 human mantle cell lymphoma cells.

[0758] 11. Inhibitory effect of compounds on MDA-MB-453 cell proliferation (1) Cell Plating: First, tumor cells MDA-MB-453 were cultured in the corresponding medium, then digested with trypsin, centrifuged, resuspended, counted, and adjusted to a concentration suitable for plating. The sources of the tumor cells are listed in Table 21. [Table 67]

[0759] Co-incubation of the compound of the present invention and tumor cells: After the cells adhered to the wall, the culture medium was removed, and the diluted bioactive molecule (compound of the present invention) was added to the wells of the plate and incubated for 72 hours.

[0760] In vitro cell viability detection: After incubation, 50 μL of CellCounting-Lite® 2.0 reagent (Vazyme / Novazyme) was added to each well, shaken, thoroughly mixed in the dark, and allowed to react for 10 minutes. Then, detection was performed using a microplate reader (manufacturer: BMG, model: PHERAStar-FS) for reading. Background RLU was obtained using cell-free medium (CellCounting-Lite®), and vehicle RLU was obtained using cell-containing medium (CellCounting-Lite®). Cell inhibition rate = 1 - (sample RLU - sample RLU) / (solvent RLU - background RLU) × 100%. The maximum inhibitory concentration (IC50) of the compound at half volume was calculated according to a 4-parameter model fitted curve, and the detection results are shown in Table 22.

[0761] (2) Data results [Table 68]

[0762] The detection results showed that the compounds of the present invention shown in Table 22 had a significant inhibitory effect on the proliferation of MDA-MB-453 human breast cancer cells.

[0763] The structures of comparative compound 1 and comparative compound 2 were as follows: [ka]

[0764] 2. Antibody conjugation test The preparation and conjugation of the ADC DL-15 sample were as follows: 1.036 mL of hIgG antibody (anti-chicken lysozyme antibody, 19.3 mg / mL) was diluted with 0.1 M disodium edetate solution (pH 7.6), then the pH was adjusted to 7.6 using 1 M Na2HPO4 solution. 2.4 times the amount of the substance was added to 10 mM TCEP (tris(2-carboxyethyl)phosphine) solution (pH 7.6), and the mixture was thoroughly mixed and allowed to stand at room temperature for 90 minutes. Compound DL-15 dissolved in 5 times the amount of the substance was added to the above solution system, and the mixture was thoroughly mixed and allowed to stand at room temperature for 2 hours. The buffer was then replaced with 10 mM histidine buffer solution at pH 6.0 using a NAP-5 gel column (Cytiva), and then sucrose and Tween-20 were added and thoroughly mixed to obtain the antibody-drug conjugate ADC DL-15 (1.77 mL, 8.60 mg / mL). [ka]

[0765] As shown in Tables 23 and 24, the molecular weight of ADC DL-15 was determined by LC-MS, and the calculated drug-to-antibody ratio, i.e., the DAR value, was 4.11.

[0766] [Table 69] [Table 70]

[0767] Chromatography determination conditions: Liquid chromatography column: Thermo MAbPac RP 3.0*100mm; Mobile phase A: 0.1%FA / H2O; Mobile phase B: 0.1%FA / ACN; Flow rate: 0.25 mL / min; Sample chamber temperature: 8°C; Column temperature: 60°C; Injection volume: 2 μL; [Table 71]

[0768] Mass spectrometry conditions: Mass spectrometer model: AB Sciex Triple TOF 5600+; GS1 35; GS2 35; CUR 30; TEM 350; ISVF 5500; DP 250; CE 10; Storage time 0.5 seconds; m / z 600~4000; Total time bin 40.

[0769] The preparation and conjugation of the ADC 3-4-04-A sample were as follows: 0.518 mL of hIgG antibody (anti-chicken lysozyme antibody, 19.3 mg / mL) was diluted with 0.1 M disodium edetate solution (pH 7.6), then the pH was adjusted to 7.6 using 1 M Na2HPO4 solution. 5.5 times the amount of the antibody was then added in 10 mM TCEP (tris(2-carboxyethyl)phosphine) solution (pH 7.6), mixed thoroughly, and allowed to stand at room temperature for 90 minutes. To the above solution system, compound 3-4-04-A dissolved in dimethyl sulfoxide in an amount 10 times the amount of the substance was added, mixed thoroughly, and allowed to stand at room temperature for 2 hours. The buffer was then replaced with 10 mM histidine buffer solution at pH 6.0 using a NAP-5 gel column (Cytiva), and then sucrose and Tween-20 were added and mixed thoroughly to obtain antibody-drug conjugate ADC 3-4-04-A (1.50 mL, 5.60 mg / mL). [ka]

[0770] As shown in Tables 25 and 26, the molecular weight of ADC 3-4-04-A was determined by LC-MS, and the calculated drug-to-antibody ratio, i.e., the DAR value, was 7.47. The chromatographic determination conditions were the same as those for ADC DL-15.

[0771] [Table 72] [Table 73]

[0772] The preparation and conjugation of the ADC 3-4-04-B sample were as follows: 0.518 mL of hIgG antibody (anti-chicken lysozyme antibody, 19.3 mg / mL) was diluted with 0.1 M disodium edetate solution (pH 7.6), then the pH was adjusted to 7.6 using 1 M Na2HPO4 solution. 5.5 times the amount of the antibody was then added in 10 mM TCEP (tris(2-carboxyethyl)phosphine) solution (pH 7.6), mixed thoroughly, and allowed to stand at room temperature for 90 minutes. To the above solution system, compound 3-4-04-B dissolved in dimethyl sulfoxide in an amount 10 times the amount of the substance was added, mixed thoroughly, and allowed to stand at room temperature for 2 hours. The buffer was then replaced with 10 mM histidine buffer solution at pH 6.0 using a NAP-5 gel column (Cytiva), and then sucrose and Tween-20 were added and mixed thoroughly to obtain antibody-drug conjugate ADC 3-4-04-B (1.50 mL, 5.60 mg / mL). [ka]

[0773] As shown in Tables 27 and 28, the molecular weight of ADC 3-4-04-B was determined by LC-MS, and the calculated drug-to-antibody ratio, i.e., the DAR value, was 8.04. The chromatographic determination conditions were the same as those for ADC DL-15.

[0774] [Table 74] [Table 75]

[0775] The above experiment demonstrated that the cytotoxic drug-linker compound of the present invention was successfully conjugated with an antibody to obtain an antibody-drug conjugate.

[0776] While specific implementations of the present invention have been described in detail, those skilled in the art will understand that various modifications and substitutions can be made to these details in accordance with all disclosed teachings, and that all such changes remain within the scope of the protection of the present invention. The full scope of the present invention is shown by the appended claims and their equivalents.

Claims

1. A pharmaceutical composition for use in the treatment of diseases related to abnormal cell proliferation, The following compounds, or their pharmaceutically acceptable salts, stereoisomers, solvates, or nitrogen oxides, and one or more pharmaceutically acceptable carriers, The diseases associated with the aforementioned abnormal cell proliferation are selected from the group consisting of brain tumors, lung cancer, squamous cell carcinoma, bladder cancer, stomach cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, cancers of the female reproductive system, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colorectal cancer, testicular cancer, gastrointestinal stromal tumors, prostate tumors, mast cell tumors, multiple myeloma, melanoma, glioma, or sarcoma. The compound has the structure of formula (II)-1: 【Chemistry 1】 (In the formula, R x' is hydrogen, R y' is hydrogen, and R z' is selected from the group consisting of hydrogen, C1-6 alkyl and C3-6 cycloalkyl, or R y' and R z' are bonded to adjacent carbon atoms to form a 3-6 membered cycloalkyl. A pharmaceutical composition having the following characteristics.

2. The compound has the following structure: 【Chemistry 2】 A pharmaceutical composition according to claim 1, having the following characteristics.

3. The use of the following compounds, or their pharmaceutically acceptable salts, stereoisomers, solvates, or nitrogen oxides, in the manufacture of pharmaceuticals for the treatment of diseases associated with abnormal cell proliferation: The diseases associated with the aforementioned abnormal cell proliferation are selected from the group consisting of brain tumors, lung cancer, squamous cell carcinoma, bladder cancer, stomach cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, cancers of the female reproductive system, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colorectal cancer, testicular cancer, gastrointestinal stromal tumors, prostate tumors, mast cell tumors, multiple myeloma, melanoma, glioma, or sarcoma. The compound has the structure of formula (II)-1: 【Transformation 3】 (In the formula, R x' is hydrogen, R y' is hydrogen, and R z' is selected from the group consisting of hydrogen, C1-6 alkyl and C3-6 cycloalkyl, or R y' and R z' are bonded to adjacent carbon atoms to form a 3-6 membered cycloalkyl. To have, to use.

4. The compound has the following structure: 【Chemistry 4】 The use according to claim 3, having the following characteristics.

5. A compound comprising 0.01 mg to 1000 mg of the following compounds, or a pharmaceutically acceptable salt, stereoisomer, solvate, or nitrogen oxide thereof, and one or more pharmaceutically acceptable carriers, The compound has the structure of formula (II)-1: 【Transformation 5】 (In the formula, R x' is hydrogen, R y' is hydrogen, and R z' is selected from the group consisting of hydrogen, C1-6 alkyl and C3-6 cycloalkyl, or R y' and R z' are bonded to adjacent carbon atoms to form a 3-6 membered cycloalkyl. A pharmaceutical composition having the following characteristics.

6. The compound has the following structure: 【Transformation 6】 A pharmaceutical composition according to claim 5, having the following characteristics.

7. A pharmaceutical composition for use in the treatment of diseases related to abnormal cell proliferation, The following compounds, or their pharmaceutically acceptable salts, stereoisomers, solvates, or nitrogen oxides, and one or more pharmaceutically acceptable carriers, The diseases associated with the aforementioned abnormal cell proliferation are selected from the group consisting of brain tumors, lung cancer, squamous cell carcinoma, bladder cancer, stomach cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, cancers of the female reproductive system, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colorectal cancer, testicular cancer, gastrointestinal stromal tumors, prostate tumors, mast cell tumors, multiple myeloma, melanoma, glioma, or sarcoma. The compound has the structure of formula (VI): M-L-E-D Equation (VI) (In the formula, D is selected from structures formed by removing a hydrogen atom from the compound described in claim 1 or 2; M has the following structure: 【Transformation 7】 Selected from the group consisting of; L is C1-6 alkylene, -N(R')-, carbonyl, -O-, Val, Cit, Phe, Lys, D-Val, Leu, Gly, Ala, Asn, Val-Cit, Val-Ala, Val-Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-Leu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, Ala-Ala-Ala, Val-Lys-Ala, Gly-Gly-Gly, Gly-Gly-Phe-Gly (Sequence ID 1), Gly-Gly-Gly-Gly-Gly (Sequence ID 2), 【Transformation 8】 (In the formula, R' represents hydrogen, C1-6 alkyl, or -(CH2CH2O)r- containing alkyl; r is an integer selected from 1 to 10; and s is an integer selected from 1 to 10.) It is a divalent structure composed of one or more elements selected from the group consisting of; E is a single bond, -NH-CH2-, 【Chemistry 9】 (Selected from the group consisting of) A pharmaceutical composition having the following characteristics.

8. The compound has the structure shown below: 【Chemistry 10】 A pharmaceutical composition according to claim 7, having the following characteristics.

9. The use of the following compounds, or their pharmaceutically acceptable salts, stereoisomers, solvates, or nitrogen oxides, in the manufacture of pharmaceuticals for the treatment of diseases associated with abnormal cell proliferation: The diseases associated with the aforementioned abnormal cell proliferation are selected from the group consisting of brain tumors, lung cancer, squamous cell carcinoma, bladder cancer, stomach cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, cancers of the female reproductive system, carcinoma in situ, lymphoma, neurofibroma, thyroid cancer, bone cancer, skin cancer, brain cancer, colorectal cancer, testicular cancer, gastrointestinal stromal tumors, prostate tumors, mast cell tumors, multiple myeloma, melanoma, glioma, or sarcoma. The compound has the structure of formula (VI): M-L-E-D Equation (VI) (In the formula, D is selected from structures formed by removing a hydrogen atom from the compound described in claim 1 or 2; M has the following structure: 【Chemistry 11】 Selected from the group consisting of; L is C1-6 alkylene, -N(R')-, carbonyl, -O-, Val, Cit, Phe, Lys, D-Val, Leu, Gly, Ala, Asn, Val-Cit, Val-Ala, Val-Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-Leu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, Ala-Ala-Ala, Val-Lys-Ala, Gly-Gly-Gly, Gly-Gly-Phe-Gly (Sequence ID 1), Gly-Gly-Gly-Gly-Gly (Sequence ID 2), 【Chemistry 12】 (In the formula, R' represents hydrogen, C1-6 alkyl, or -(CH2CH2O)r- containing alkyl; r is an integer selected from 1 to 10; and s is an integer selected from 1 to 10.) It is a divalent structure composed of one or more elements selected from the group consisting of; E is a single bond, -NH-CH2-, 【Chemistry 13】 (Selected from the group consisting of) To have, to use.

10. The compound has the structure shown below: 【Chemistry 14】 The use according to claim 9, having the following characteristics.

11. A compound comprising 0.01 mg to 1000 mg of the following compounds, or a pharmaceutically acceptable salt, stereoisomer, solvate, or nitrogen oxide thereof, and one or more pharmaceutically acceptable carriers, The compound has the structure of formula (VI): M-L-E-D Equation (VI) (In the formula, D is selected from structures formed by removing a hydrogen atom from the compound described in claim 1 or 2; M has the following structure: 【Chemistry 15】 Selected from the group consisting of; L is C1-6 alkylene, -N(R')-, carbonyl, -O-, Val, Cit, Phe, Lys, D-Val, Leu, Gly, Ala, Asn, Val-Cit, Val-Ala, Val-Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-Leu-Lys, Gly-Gly-Arg, Ala-Ala-Asn, Ala-Ala-Ala, Val-Lys-Ala, Gly-Gly-Gly, Gly-Gly-Phe-Gly (Sequence ID 1), Gly-Gly-Gly-Gly-Gly (Sequence ID 2), 【Chemistry 16】 (In the formula, R' represents hydrogen, C1-6 alkyl, or -(CH2CH2O)r- containing alkyl; r is an integer selected from 1 to 10; and s is an integer selected from 1 to 10.) It is a divalent structure composed of one or more elements selected from the group consisting of; E is a single bond, -NH-CH2-, 【Chemistry 17】 (Selected from the group consisting of) A pharmaceutical composition having the following characteristics.

12. The compound has the structure shown below: [Chemistry 18] A pharmaceutical composition according to claim 11, having the following characteristics.

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