Camptothecin derivatives, pharmaceutical compositions, methods for producing the same, and applications

Abstract

The present invention relates to a camptothecin derivative represented by formula I, a pharmaceutical composition, and a method for producing and using the same. The camptothecin derivative has good tumor inhibitory activity and can be used to produce pharmaceuticals for the treatment or prevention of tumors and for the treatment or prevention of neoplastic diseases. TIFF2026513781000319.tif52108

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to six applications filed on April 7, 2023 (application number 202310367781.7), June 21, 2023 (application number 202310743291.2), July 28, 2023 (application number 202310941713.7), September 12, 2023 (application number 202311173712.9), November 21, 2023 (application number 202311558824.6), and December 25, 2023 (application number 202311802330.8), all of which are titled "Camptothecin derivatives, pharmaceutical compositions, and methods and uses thereof."

[0002] This invention belongs to the pharmaceutical field and specifically relates to camptothecin derivatives, pharmaceutical compositions, and methods for producing and using the same. [Background technology]

[0003] In the field of tumor therapy, antibody-drug conjugates (ADCs) are constructed by conjugating a monoclonal antibody drug that targets a specific antigen with a small molecule cytotoxic drug via a connector, possessing both the potent killing effect of conventional small molecule chemotherapy and the tumor targeting capabilities of antibody drugs.

[0004] Camptothecin (CPT) is a natural compound isolated from the Camptotheca acuminata plant of the Cornaceae family. Camptothecin is a five-ring fused compound consisting of a quinoline A / B ring, a pyrrole C ring, a pyridinone D ring, and an α-hydroxylactone E ring, with the 20th position being in an S configuration (see structural formula below). Due to its excellent anticancer activity, it was introduced into clinical practice in the early 1970s, but clinical trials were discontinued after serious drug side effects such as diarrhea and hemorrhagic cystitis occurred in clinical settings. [ka]

[0005] Research data indicates that camptothecin can induce cell death by forming a ternary complex with DNA topoisomerase I in cells, inhibiting DNA unwinding and thus inhibiting DNA replication (Cancer Res. 1989, 49, 6365). Camptothecin and its derivatives exhibit potent antitumor activity in animal in vivo models of lung cancer, breast cancer, colorectal cancer, ovarian cancer, etc. (Nature Review Cancer. 2006, 6, 789). Currently, several camptothecin-type drugs are approved for the treatment of tumors (Med Res. Rev. 2015, 35, 753). Irinotecan is a drug 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, diflomotecan, ruthecan, gimatecan, simitecan, quimitecan, and aeromothecan. Camptothecin drugs or derivatives often cause hematological toxicity due to bone marrow suppression, such as leukopenia, thrombocytopenia, anemia, and neutropenia, as well as gastrointestinal side effects such as nausea, vomiting, and diarrhea. Clinical studies have revealed that means to improve the safety and efficacy of camptothecin compounds include increasing water solubility, improving their pharmacokinetic properties, enhancing activity, reducing the dose, or using their conjugates to form antibody-drug conjugates. Therefore, developing camptothecin compounds and their conjugates with novel structures that can improve efficacy and mitigate safety issues still has high clinical need and application value. [Overview of the project]

[0006] The present invention provides compounds represented by formula I, their racemates, stereoisomers, tautomers, isotopically labeled compounds, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof. [ka] Here, R1, R2, and R3 are the same or different and are independently of each other H, OH, CN, halogen, C 1~10 alkyl group, C 2~10 alkenyl group, C 2~10 alkynyl group, C 1~10 alkoxy group, halo C 1~10 alkyl group, halo C 1~10 alkoxy group, cyano C 1~10 alkyl group, cyano C 1~10 alkoxy group, C 3~10 cycloalkyl group, selected from R4 is H or

Chemical formula

Chemical formula

[0007] According to some embodiments, R1 is H, OH, CN, halogen, C 1~6 Alkyl alkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl groups or halo C 1~6 Selected from alkoxy groups. According to some embodiments, R1 is selected from H, OH, Br, a methyl group, a difluoromethoxy group, a 2,2,2-trifluoroethoxy group, a vinyl group, a cyclopropyl group, or an ethynyl group.

[0008] According to some embodiments, R2 is H, halogen, CN, or C 1~6 Selected from alkyl groups. According to some embodiments, R2 is selected from H or F.

[0009] According to some embodiments, R3 is H or C 1~6 Selected from alkyl groups. According to some embodiments, R3 is H.

[0010] According to some embodiments, R4 is H or [ka] Selected from, for example [ka] That is the case. According to some embodiments, R4 is H or [ka] Selected from.

[0011] According to some embodiments, the X-R4 [ka] Preferably [ka] That is the case.

[0012] According to some embodiments, X-R4 is -CH2-.

[0013] According to some embodiments, R5 is H, [ka] Selected from, R 51 R is selected from H, methyl group, ethyl group, isopropyl group or cyclopropyl group, 52 R is selected from H or methyl group, 53 The group is selected from methyl groups, and the A ring is C 3~6 Selected from cycloalkyl groups, Ra is H, hydroxyl group, CN, halogen, C 1~6 Alkyl alkyl group, C 1~6 Selected from a haloalkyl group, n is selected from 0 or 1, and q is selected from 0 or 1. According to some embodiments, ring A is selected from cyclobutane rings.

[0014] According to some embodiments, R5 is H, [ka] Selected from. According to some embodiments, R 51 R is selected from H, methyl group, ethyl group, isopropyl group or cyclopropyl group, 52 R is selected from H or methyl group, 53 The A ring is selected from methyl groups, and the A ring is selected from cyclobutane rings. According to some embodiments, R5 is H, [ka] Selected from.

[0015] According to some embodiments, X is selected from CH or N, and if X is CH, R4 is H, or if X is N, R5 is selected from H. According to some embodiments, m is selected from 0, 1, or 2. According to some embodiments, p is selected from 0, 1, or 2. According to some examples, Y is selected from -CH2-, -CH2-CH2-, and -CH2-O-.

[0016] According to some embodiments, the structure of the compound represented by formula I is as follows: [ka] Here, R1, R2, R3, R4, R5, X, and m have the definitions set forth herein, independently of each other.

[0017] According to some embodiments, the structure of the compound represented by formula I is as follows: [ka] R1, R2, R4, R5, X, and m have the definitions set forth herein, independently of each other.

[0018] According to some embodiments, the structure of the compound represented by formula I is as follows: [ka] Here, R1, R2, and R5 have the definitions set forth herein, independently of each other.

[0019] According to some embodiments, the structure of the compound represented by formula I is as follows: [ka] Here, R1, R2, R 51 , R 52 Ring A, Ra, m, n, and q have the definitions set forth herein, independently of each other.

[0020] According to embodiments of the present invention, the structure of compound I is [ka] [ka] [ka] [ka] [ka] [ka] As shown.

[0021] The present invention further provides compounds represented by formula V, their racemates, stereoisomers, tautomers, isotopic markers, solvates, polytypes, pharmaceutically acceptable salts, or prodrug compounds thereof. M-L1-L2-D(Formula V) Here, M is the linker site with the antibody or its antigen-binding fragment. L1 is a peptide residue, preferably selected from glycine-glycine-phenylalanine-glycine (GGFG), glutamic acid-valine-citrulline (EVC), valine-citrulline (VC), aspartic acid-valine-citrulline (DVC), glutamic acid-glycine-glycine-phenylalanine-glycine (EGGFG), and aspartic acid-glycine-glycine-phenylalanine-glycine (DGGFG). L2 is a linking group between the peptide residue and D, preferably a chemical bond, -NH-C 1~6 Alkyl-, [ka] Selected from, preferably L2 is selected from -NH-CH2-, D is a structural fragment of a biologically active molecule.

[0022] According to some embodiments, D is selected from the dehydrogenated compound structures represented by Formula I described herein.

[0023] According to some embodiments, the structure of D is [ka]

Chem.

Chem.

Chem.

Chem.

Chem.

[0024] According to some embodiments, M is selected from the following structures,

Chem.

[0025] According to some embodiments, ring B is selected from a 5- to 6-membered N-containing heteroaryl ring, a 3- to 6-membered N-containing heterocyclic ring According to some embodiments, ring B is a pyrimidine ring, a pyridine ring, a triazine ring [Chemical formula] and is selected from According to some embodiments, each R b are the same or different and are independently of each other a cyano group, oxo (=O), methoxy group, cyclopropyl group, trifluoromethyl group [Chemical formula] and is selected from According to some embodiments, L m1 is absent, or is unsubstituted or is substituted with one, two or more R m1The group that is optionally substituted is selected from a phenyl group, a piperidinyl group, or a piperazinyl group. According to some embodiments, R m1 It is selected from -CH2COOH. According to some embodiments, L m1 teeth [ka] Selected from. According to some embodiments, L m2 This is either no substitution or one, two, or more R m2 The bases selected are -CH2-(C=O)-, -(CH2)2-(C=O)-, -(CH2)5-(C=O)-, and -C≡C-(CH2)3-(C=O)-, which are arbitrarily substituted. According to some embodiments, each R m2 They are either the same or different, and are independent of each other -C 1~3 Selected from alkylene-COOH, the alkylene group is optionally interrupted by O or NH. According to some embodiments, each R m2 They are either the same or different, and independently of each other. [ka] Selected from. According to some embodiments, M is [ka] [ka] Selected from. According to some embodiments, L1 is glycine-glycine-phenylalanine-glycine (GGFG), i.e. [ka] That is the case. According to some embodiments, L2 is -NH-CH2-.

[0026] According to some embodiments, M-L1-L2 is [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] selected from the group consisting of.

[0027] According to some embodiments, the compound shown in Formula V is [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] It is selected from this structure.

[0028] The present invention further provides an antibody-drug conjugate represented by formula VI, Ab-[LD] β (Formula VI) Here, Ab is an antibody or its antigen-binding fragment, D has the definition described in this specification, L is a connector linking Ab and D, and β is selected from an integer or decimal number between 1 and 10.

[0029] According to some embodiments, Ab is an antibody or antigen-binding fragment, the antigen-binding fragment being selected from Fab, Fab', (Fab')2, Fd, Fv, Fv linked by disulfide bonds, scFv, di-scFv, (scFv)2, diabody, and single-domain antibody (sdAb), and / or the antibody is a mouse-derived antibody, a humanized antibody, a chimeric antibody, a bispecific antibody, or a multispecific antibody.

[0030] According to some embodiments, Ab is an anti-HER2 antibody or its antigen-binding fragment, for example, Ab is trastuzumab or its antigen-binding fragment.

[0031] According to some embodiments, β is selected from integers or decimals between 4 and 9 (e.g., 7, 7.71, 7.84, 7.92, 7.94, 7.97, 7.98, 7.99, 8, 8.02, 8.06, or 8.14).

[0032] According to some embodiments, L is selected from M'-L1-L2, where M' is a linker site with an antibody or its antigen-binding fragment, formed by the complexation of M as defined in this specification with an antibody or its antigen-binding fragment, and L1, L2 have the definitions defined in this specification. According to some embodiments, M' is [ka] [ka] Selected from.

[0033] Preferably, the carbonyl group linkage position in M' is linked to L1, and the linkage position in the heterocyclic or heteroaryl ring is linked to Ab.

[0034] According to some embodiments, L is selected from the following: [ka] [ka] [ka] [ka] [ka] Here, the first-place entry is linked to Ab, and the second-place entry is linked to D.

[0035] According to some embodiments, the LD is [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] Selected from.

[0036] According to some embodiments, the antibody-drug conjugate represented by formula VI is [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] It is selected from this structure.

[0037] The present invention further provides a method for producing a compound of formula I, comprising the following methods 1 and 2: Method 1 is, Step (1) removes the protecting group PG4 from compound I-41 to obtain compound I-42, The process includes step (2) reacting compound I-42 with compound I-43 to obtain the compound shown in formula I, [ka] Here, R1, R2, R3, R4, R5, X, and m have the definitions set forth herein, Y is a leaving group selected from, for example, OH, Cl, Br, and I, and PG4 is an amino protecting group selected from, for example, Fmoc, Boc, Bn, and Cbz.

[0038] Method 2 is, Step (1) removes the protecting group PG5 from compound I-51 to obtain compound I-52, The process includes step (2) reacting compound I-52 with compound I-53 to obtain the compound shown in formula I, [ka] Here, R1, R2, R3, R4, R5, R 51 X, m, and n have the definitions set forth herein, Y is a leaving group selected from, for example, OH, Cl, Br, and I, and PG5 is an amino protecting group selected from, for example, Fmoc, Boc, Bn, and Cbz.

[0039] The present invention further provides a pharmaceutical composition comprising at least one of the following: a therapeutically effective amount of a compound represented by formula I or formula V, its racemic mixture, stereoisomer, tautomer, isotopic marker, solvate, polytype, pharmaceutically acceptable salt, or prodrug compound thereof.

[0040] The present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of an antibody-drug conjugate represented by formula VI.

[0041] According to embodiments of the present invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable auxiliary materials.

[0042] According to embodiments of the present invention, the pharmaceutical composition may further comprise one or more additional therapeutic agents.

[0043] The present invention further provides a method for treating neoplastic diseases, comprising administering to a patient at least one of the compounds represented by formula I or formula V, their racemates, stereoisomers, tautomers, isotopic markers, solvates, polytypes, pharmaceutically acceptable salts, or prodrug compounds thereof in a prophylactic or therapeutically effective amount.

[0044] The present invention further provides a method for treating neoplastic diseases, which comprises administering a preventive or therapeutically effective amount of the above-mentioned pharmaceutical composition to a patient.

[0045] The aforementioned neoplastic disease is selected from breast cancer, gastric cancer, lung cancer, colorectal cancer, colon cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal cancer, melanoma, or leukemia.

[0046] In some embodiments, the patient includes mammals, preferably humans.

[0047] The present invention further provides at least one of the compounds represented by formulas I, V, or antibody-drug conjugates represented by formula VI, racemates, stereoisomers, tautomers, isotope-labeled compounds, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof, or a pharmaceutical composition thereof, used for treating neoplastic diseases.

[0048] The present invention further provides the use of at least one of the compounds represented by formulas I, V, or antibody-drug conjugates represented by formula VI, their racemates, stereoisomers, tautomers, isotope-labeled, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds, or the above-mentioned pharmaceutical compositions, in the manufacture of topoisomerase I inhibitors and / or in the manufacture of pharmaceuticals for the prevention or treatment of diseases or disorders related to topoisomerase I.

[0049] In some embodiments, the disease or disorder is a tumor, and the tumor includes breast cancer, gastric cancer, lung cancer, colorectal cancer, colon cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal cancer, melanoma, or leukemia.

[0050] The present invention further provides the compound represented by formula I', its racemic mixture, stereoisomer, tautomer, isotopic marker, solvate, polytype, pharmaceutically acceptable salt, or prodrug compound thereof. [ka] Here, R1, R2, and R3 are either the same or different, and independently of each other, they are H, OH, CN, halogen, and C. 1~10 Alkyl alkyl group, C 2~10Alkenyl group, C 2~10 Alkynyl group, C 1~10 Alkoxy group, Halo C 1~10 Alkyl, halo C 1~10 Alkoxy group, cyanoC 1~10 Alkyl alkyl, cyano C 1~10 Alkoxy group, C 3~10 Selected from cycloalkyl groups, R4 is H or [ka] Selected from, R 41 H, C 1~6 Alkyl alkyl group, C 1~6 Haloalkyl group, C 1~6 Alkyl-NH-, (C 1~6 Alkyl)2N-,C 1~6 Alkyl-NH-C 1~6 Alkyl, (C 1~6 Alkyl)2N-C 1~6 Alkyl alkyl group, C 1~6 Alkylalkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl group, 3-6 membered heterocyclyl group, C 6~14 Selected from aryl groups and 5-14 membered heteroaryl groups, R5 is H, [ka] Selected from, R 51 , R 52 H and C are either the same or different, and are independent of each other. 1~6 Alkyl alkyl group, C 1~6 Haloalkyl group, C 1~6 Alkyl-NH-, (C 1~6 Alkyl)2N-,C 1~6 Alkyl-NH-C 1~6 Alkyl, (C 1~6 Alkyl)2N-C 1~6 Alkyl alkyl group, C 1~6 Alkylalkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6Cycloalkyl group, 3-6 membered heterocyclyl group, C 6~14 Selected from aryl groups and 5-14 membered heteroaryl groups, R 53 is C 1~6 Alkyl alkyl group, C 1~6 Haloalkyl group, C 1~6 Alkyl-NH-, (C 1~6 Alkyl)2N-,C 1~6 Alkyl-NH-C 1~6 Alkyl, (C 1~6 Alkyl)2N-C 1~6 Alkyl alkyl group, C 1~6 Alkylalkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl group, 3-6 membered heterocyclyl group, C 6~14 Selected from aryl groups and 5-14 membered heteroaryl groups, the A ring is C 3~8 Cycloalkyl groups or C 3~8 Selected from heterocyclyl groups, Ra is H, hydroxyl group, CN, halogen, C 1~6 Alkyl alkyl group, C 1~6 Selected from haloalkyl groups, n is selected from 0, 1, or 2, and q is selected from 0, 1, or 2. X is selected from CH or N. Y is -(CH2) m -O-(CH2) p - Selected from, m is selected from integers between 0 and 6. p is selected from integers between 0 and 6.

[0051] According to some embodiments, R1 is H, OH, CN, halogen, C 1~6 Alkyl alkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl groups or halo C 1~6 Selected from alkoxy groups, Preferably, R1 is selected from H, OH, Br, methyl group, difluoromethoxy group, 2,2,2-trifluoroethoxy, vinyl group, cyclopropyl group or ethynyl group. Preferably, R2 is H, halogen, CN, or C 1~6 Selected from alkyl groups, Preferably, R2 is selected from H or F. Preferably, R3 is H or C 1~6 Selected from alkyl groups, Preferably, R3 is H.

[0052] According to some embodiments, R4 is H or [ka] Selected from, for example [ka] Selected from, Preferably, R4 is H or [ka] Selected from, Preferably, the X-R4 is [ka] Preferably [ka] And, Preferably, X-R4 is -CH2-, Preferably, Y is selected from -CH2-O-.

[0053] According to some embodiments, R5 is H, [ka] Selected from, R 51 R is selected from H, methyl group, ethyl group, isopropyl group or cyclopropyl group, 52 R is selected from H or methyl group, 53 The group is selected from methyl groups, and the A ring is C 3~6Selected from cycloalkyl groups, Ra is H, hydroxyl group, CN, halogen, C 1~6 Alkyl alkyl group, C 1~6 Selected from haloalkyl groups, n is selected from 0 or 1, and q is selected from 0 or 1. Preferably, the A ring is selected from a cyclobutane ring. Preferably, R5 is H, [ka] Selected from, Preferably, R 51 R is selected from H, methyl group, ethyl group, isopropyl group or cyclopropyl group, 52 R is selected from H or methyl group, 53 The A ring is selected from a methyl group, and the A ring is selected from a cyclobutane ring. Preferably, R5 is H, [ka] Selected from.

[0054] According to some embodiments, X is selected from CH or N, and if X is CH, R4 is H, or if X is N, R5 is selected from H. Preferably, m is selected from 0, 1, or 2, and more preferably, m+p is 2.

[0055] According to some embodiments, the structure of the compound represented by formula I' is as follows: [ka] Here, R1, R2, R 51 , R 52 n, independently of each other, has the definitions described above.

[0056] According to some embodiments, the structure of the compound shown in formula I' is, [ka] [ka] As shown.

[0057] The present invention further provides compounds represented by formula I'', their racemates, stereoisomers, tautomers, isotopic markers, solvates, polytypes, pharmaceutically acceptable salts, or prodrug compounds thereof. [ka] Here, R1, R2, and R3 are either the same or different, and independently of each other, they are H, OH, CN, halogen, and C. 1~10 Alkyl alkyl group, C 2~10 Alkenyl group, C 2~10 Alkynyl group, C 1~10 Alkoxy group, Halo C 1~10 Alkyl, halo C 1~10 Alkoxy group, cyanoC 1~10 Alkyl alkyl, cyano C 1~10 Alkoxy group, C 3~10 Selected from cycloalkyl groups, R4 is H or [ka] Selected from, R 41 H, C 1~6 Alkyl alkyl group, C 1~6 Haloalkyl group, C 1~6 Alkyl-NH-, (C 1~6 Alkyl)2N-,C 1~6 Alkyl-NH-C 1~6 Alkyl, (C 1~6 Alkyl)2N-C 1~6 Alkyl alkyl group, C 1~6 Alkylalkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl group, 3-6 membered heterocyclyl group, C 6~14 Selected from aryl groups and 5-14 membered heteroaryl groups, R 52 H, C 1~6 Alkyl alkyl group, C 1~6Haloalkyl group, C 1~6 Alkyl-NH-, (C 1~6 Alkyl)2N-,C 1~6 Alkyl-NH-C 1~6 Alkyl, (C 1~6 Alkyl)2N-C 1~6 Alkyl alkyl group, C 1~6 Alkylalkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl group, 3-6 membered heterocyclyl group, C 6~14 Selected from aryl groups and 5-14 membered heteroaryl groups, R 20 is selected from H or an amino protecting group, Preferably, R 20 The following can be selected from Fmoc, Boc, Bn, and Cbz: X is selected from CH or N. Y is -(CH2) m -O-(CH2) p - Selected from, m is selected from integers between 0 and 6. p is selected from integers between 0 and 6. Preferably, R1 is H, OH, CN, halogen, C 1~6 Alkyl alkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl groups or halo C 1~6 Selected from alkoxy groups, Preferably, R1 is selected from H, OH, Br, methyl group, difluoromethoxy group, 2,2,2-trifluoroethoxy, vinyl group, cyclopropyl group or ethynyl group. Preferably, R2 is H, halogen, CN, or C 1~6 Selected from alkyl groups, Preferably, R2 is selected from H or F. Preferably, R3 is H or C 1~6 Selected from alkyl groups, Preferably, R3 is H, R4 is selected from H, Preferably, formula I'' is the following compound: [ka] [ka] That is the case.

[0058] The present invention further provides the compound represented by formula MII, [ka] Lg is a leaving group, and the leaving group is selected from a halogen, a sulfonyl group, a trifluoromethanesulfonyl group, or a methylsulfonyl group, preferably Lg is a methylsulfonyl group. Ring B is selected from a 5-6 membered nitrogen-containing heteroaryl ring or a 3-6 membered nitrogen-containing heteroring. More preferably, the B ring is selected from a pyrimidine ring, a pyridine ring, or a triazine ring. Each R b They are either the same or different, and independently of each other are halogens, cyano groups, oxo (=O), and C 1~6 Alkyl, halo C 1~6 Alkyl alkyl, hydroxy C 1~6 Alkyl alkyl group, C 1~6 Alkoxy group or C 3~8 Cycloalkyl group, 3-8 membered heterocyclyl group, C 1~6 Alkyl-OC 1~6 Alkyl-, C 1~6 Selected from alkyl-(5-6 member) heteroaryl- groups, Preferably, each R b These are either the same or different, and independently of each other are cyano, oxo (=O), methoxy, cyclopropyl, and trifluoromethyl groups. [ka] Selected from, L m1 It either does not exist, is unsubstituted, or has one, two, or more R values. m1 C is optionally replaced by 6~14The groups selected are aryl groups, 5-14 membered heteroaryl groups, and 3-14 membered heterocyclyl groups, and each R m1 They are either the same or different, and independently of each other are H, halogen, cyano group, and C 1~6 Alkyl alkyl group, HOOC-C 1~3 Selected from alkylene groups, Preferably, L m1 It either does not exist, is unsubstituted, or has one, two, or more R values. m1 The group that is optionally substituted is selected from a phenyl group, a piperidinyl group, or a piperazinyl group. More preferably, L m1 teeth, [ka] Selected from, R m20 This is either no substitution or one, two, or more R m2 -(CH2) is optionally substituted. s -(C=O)-R Z -ethynyl-(CH2) t -(C=O)-R Z Selected from the basis, Preferably, R m20 This is either no substitution or one, two, or more R m2 -CH2-(C=O)-R is optionally substituted. Z -(CH2)2-(C=O)-R Z -(CH2)5-(C=O)-R Z ,-ethynyl-(CH2)3-(C=O)-R Z Selected from the basis, Each R m2 They are either the same or different, and independently of each other are H, halogen, cyano group, and C 1~6 Alkyl alkyl group or -C 1~6 Selected from alkylene-COOH, the alkylene group is optionally interrupted by one, two, or more O, NH groups. Preferably, each R m2 They are either the same or different, and are independent of each other -C1~3 Selected from alkylene-COOH, the alkylene group is optionally interrupted by O or NH. Comfortable, R m2 teeth, [ka] Selected from, r is selected from integers between 0 and 6. s is selected from integers between 0 and 6. t is selected from integers between 0 and 6. Rz is selected from a hydroxyl group, halogen, active ester, carboxyl protecting group, amino acid, peptide fragment, or hydrophilic fragment. The aforementioned amino acid is the N-terminal amino acid of L1, and the peptide fragment is a sub-fragment formed from 2, 3, or 4 amino acids at the N-terminus of L1, or is L1 itself. The C-terminus of the peptide fragment is a hydroxyl group, an active ester, a carboxyl protecting group, or [ka] And, Preferably, the hydrophilic fragment comprises a polyhydroxy group, a polyethylene glycol fragment, a betaine fragment, or a polycreatine fragment. Preferably, formula MII is formula MII-1 below, [ka] Here, t and Rz are as defined above, and Z is N or CR 22 And R 21 and R 22 These are independently H, halogen, cyano group, oxo (=O), and C. 1~6 Alkyl, halo C 1~6 Alkyl alkyl, hydroxy C 1~6 Alkyl alkyl group, C 1~6 Alkoxy group, C 3~8 Cycloalkyl group, 3-8 membered heterocyclyl group, C 1~6 Alkyl-OC 1~6Selected from alkyl groups, with the condition that Z is N, R 21 It is not H, Preferably, Rz is OH or a halogen, t is an integer from 2 to 4, Z is N, and R 21 C is a cyano group, C 1~6 Alkoxy group, C 3~4 Cycloalkyl groups or -C 1~6 Alkyl-OC 1~6 Selected from alkyl groups, or R 21 H is and Z is CR 22 And R 22 This is a cyano group or a trifluoromethyl group. Preferably, formula MII is, [ka] This is the compound.

[0059] The present invention further provides compounds represented by formulas (L'-1)-(L'-3), [ka] Lg is a leaving group, and the leaving group is selected from a halogen, a sulfonyl group, a trifluoromethanesulfonyl group, or a methylsulfonyl group, preferably Lg is a methylsulfonyl group. Ring B is selected from a 5-6 membered nitrogen-containing heteroaryl ring or a 3-6 membered nitrogen-containing heteroring. More preferably, the B ring is selected from a pyrimidine ring, a pyridine ring, or a triazine ring. Each R b They are either the same or different, and independently of each other are halogens, cyano groups, oxo (=O), and C 1~6 Alkyl, halo C 1~6 Alkyl alkyl, hydroxy C 1~6 Alkyl alkyl group, C 1~6 Alkoxy group or C 3~8 Cycloalkyl group, 3-8 membered heterocyclyl group, C 1~6 Alkyl-OC 1~6 Alkyl-, C 1~6Selected from alkyl-(5-6 member) heteroaryl- groups, Preferably, each R b These are either the same or different, and independently of each other are cyano, oxo (=O), methoxy, cyclopropyl, and trifluoromethyl groups. [ka] Selected from, L m1 It either does not exist, is unsubstituted, or has one, two, or more R values. m1 C is optionally replaced by 6~14 The groups selected are aryl groups, 5-14 membered heteroaryl groups, and 3-14 membered heterocyclyl groups, and each R m1 They are either the same or different, and independently of each other are H, halogen, cyano group, and C 1~6 Alkyl alkyl group, HOOC-C 1~3 Selected from alkylene groups, Preferably, L m1 It either does not exist, is unsubstituted, or has one, two, or more R values. m1 The group that is optionally substituted is selected from a phenyl group, a piperidinyl group, or a piperazinyl group. More preferably, L m1 teeth, [ka] Selected from, L m2 This is either no substitution or one, two, or more R m2 -(CH2) is optionally substituted. s -(C=O)-, -ethynyl-(CH2) t Selected from the base -(C=O)-, Preferably, L m2 This is either no substitution or one, two, or more R m2 The group is selected from the following groups which can be arbitrarily substituted: -CH2-(C=O)-, -(CH2)2-(C=O)-, -(CH2)5-(C=O)-, and -ethynyl-(CH2)3-(C=O)-. Each R m2 They are either the same or different, and independently of each other are H, halogen, cyano group, and C 1~6 Alkyl alkyl group or -C 1~6 Selected from alkylene-COOH, the alkylene group is optionally interrupted by one, two, or more O, NH groups. Preferably, each R m2 They are either the same or different, and are independent of each other -C 1~3 Selected from alkylene-COOH, the alkylene group is optionally interrupted by O or NH. Comfortable, R m2 teeth, [ka] Selected from, r is selected from integers between 0 and 6. s is selected from integers between 0 and 6. t is selected from integers between 0 and 6. Rz2 is selected from a hydroxyl group, halogen, active ester, or carboxyl protecting group. Preferably, here, [ka] The fragment has the structure shown in the following formula, [ka] Here, each group has the definitions set forth herein. R 51 H and C are independent of each other. 1~6 Alkyl alkyl group, C 1~6 Haloalkyl group, C 1~6 Alkyl-NH-, (C 1~6 Alkyl)2N-,C 1~6 Alkyl-NH-C 1~6 Alkyl, (C 1~6 Alkyl)2N-C 1~6 Alkyl alkyl group, C 1~6 Alkylalkyl group, C 2~6 Alkenyl group, C 2~6Alkynyl group, C 3~6 Cycloalkyl group, 3-6 membered heterocyclyl group, C 6~14 Selected from aryl groups and 5-14 membered heteroaryl groups, R 53 C 1~6 Alkyl alkyl group, C 1~6 Haloalkyl group, C 1~6 Alkyl-NH-, (C 1~6 Alkyl)2N-,C 1~6 Alkyl-NH-C 1~6 Alkyl, (C 1~6 Alkyl)2N-C 1~6 Alkyl alkyl group, C 1~6 Alkylalkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl group, 3-6 membered heterocyclyl group, C 6~14 Selected from aryl groups and 5-14 membered heteroaryl groups, the A ring is C 3~8 Selected from cycloalkyl groups or 3-8 membered heterocyclyl groups, Ra is H, hydroxyl group, CN, halogen, C 1~6 Alkyl alkyl group, C 1~6 Selected from haloalkyl groups, n is selected from 0, 1, or 2, and q is selected from 0, 1, or 2. L1 is a peptide residue, preferably selected from glycine-glycine-phenylalanine-glycine (GGFG), glutamic acid-valine-citrulline (EVC), valine-citrulline (VC), aspartic acid-valine-citrulline (DVC), glutamic acid-glycine-glycine-phenylalanine-glycine (EGGFG), and aspartic acid-glycine-glycine-phenylalanine-glycine (DGGFG), and preferably L1 is selected from glycine-glycine-phenylalanine-glycine (GGFG). L2 is a linking group between the peptide residue and D, preferably a chemical bond, -NH-C 1~6 Alkyl-, [ka] Selected from, preferably L2 is selected from -NH-CH2-, (L'-1)-(L'-3) is preferably, [ka] [ka] That is the case.

[0060] The present invention further provides a method for synthesizing formula (L'), where L2 is -NH-CH2-, and comprises the following steps of the reaction: [ka] Here, Rz2 is a hydroxyl group, an active ester, or a carboxyl protecting group, and each of the remaining groups has the definitions set forth herein.

[0061] The present invention further provides a synthetic method for complexing intermediate compounds, comprising the first method or the second method, The first method is selected from the following steps: [ka] Rz2 is selected from a hydroxyl group, halogen, active ester, or carboxyl protecting group, and each of the remaining groups has the definitions set forth herein. The second method is, [ka] The process includes the step of reacting with the compound of formula I described above (where R5 is not H).

[0062] Beneficial effects The camptothecin-based compounds according to the present invention have good tumor suppressor activity and can be used for the treatment or prevention of cancer (such as breast cancer or gastric cancer) and for the manufacture of pharmaceuticals for the treatment or prevention of such disorders and diseases. The cytotoxic drug-connector compounds according to the present invention can be smoothly conjugated with antibodies to obtain antibody-drug conjugates. The antibody-drug conjugates according to the present invention have good tumor suppressor activity and selectivity.

[0063] Definitions and explanations of terms Unless otherwise specified, the definitions of groups and terms described in the specification and claims of this application may be combined in any way, including example definitions, illustrative definitions, preferred definitions, definitions in tables, and definitions of specific compounds in examples. Such combined definitions of groups and compound structures should be understood to be within the scope described in the description and / or claims of this application.

[0064] The terms "linker," "linker structure," "connector," or "linking unit" as used in this invention refer to a fragment or linkage of a chemical structure in which one end is linked to an antibody and the other end is linked to a drug (drug compound), and which may be linked to another linker before being linked to the drug compound. The linker structures of this invention may be synthesized by methods known in the art or by the methods described in this invention.

[0065] The "antibody-drug conjugate" (ADC) described in the present invention means that the target portion is linked to a biologically active drug via a stable linking unit.

[0066] The term "bioactive molecule" as used in this invention refers to a cytotoxic drug, meaning a chemical molecule that can strongly inhibit the normal growth of tumor cells.

[0067] Unless otherwise specified, numerical ranges described herein and in the claims are equivalent to listing at least the numerical values ​​of each specific integer. For example, the numerical range "1 to 12" is equivalent to listing each integer value within the numerical range "1 to 12", i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12. Furthermore, when some numerical ranges are defined as "numbers", they should be understood as listing both ends of the range, each integer within the range, and each decimal within the range.

[0068] The term "integers from 0 to 6" refers to 0, 1, 2, 3, 4, 5, and 6.

[0069] The term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0070] "C 1~10 "Alkyl alkyl" means a linear or branched alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. 1-8 "Alkyl alkyl" means a linear or branched alkyl group having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. 1~6 "Alkyl group" means a linear or branched alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, a methyl group, ethyl group, propyl group, butyl group, pentyl group, hexyl group, isopropyl group, isobutyl group, sec-butyl group, tert-butyl group, isopentyl group, 2-methylbutyl group, 1-methylbutyl group, 1-ethylpropyl group, 1,2-dimethylpropyl group, neopentyl group, 1,1-dimethylpropyl group, 4-methylpentyl group, 3-methylpentyl group, 2-methylpentyl group, 1-methylpentyl group, 2-ethylbutyl group, 1-ethylbutyl group, 3,3-dimethylbutyl group, 2,2-dimethylbutyl group, 1,1-dimethylbutyl group, 2,3-dimethylbutyl group, 1,3-dimethylbutyl group, or 1,2-dimethylbutyl group, or their isomers.

[0071] "C 2~10 The term "alkenyl group" should preferably be understood to mean a linear or branched monovalent hydrocarbon group containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, and more preferably "C 2~8 It is an "alkenyl group" "C 2~10 An "alkenyl group" contains one or more double bonds and has 2, 3, 4, 5, 6, 7, or 8 carbon atoms, for example, 2, 3, 4, 5, or 6 carbon atoms (i.e., C 2~6 Alkenyl group), having 2 or 3 carbon atoms (i.e., C 2~3The term "alkenyl group" should preferably be understood to mean a linear or branched monovalent hydrocarbon group. If the alkenyl group contains more than one double bond, the double bonds may be separated or conjugated. Examples of the alkenyl group include vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-buto-2-enyl, (Z)-buto-2-enyl, (E)-buto-1-enyl, (Z)-buto-1-enyl, pento-4-enyl, (E)-pento-3-enyl, (Z)-pento-3-enyl, (E)-pento-2-enyl, (Z)-pento-2-enyl, (E)-pento-1-enyl, (Z)-pento-1-enyl, hexa-5-enyl, (E) -Hexa-4-enyl, (Z)-Hexa-4-enyl, (E)-Hexa-3-enyl, (Z)-Hexa-3-enyl, (E)-Hexa-2-enyl, (Z)-Hexa-2-enyl, (E)-Hexa-1-enyl, (Z)-Hexa-1-enyl, Isopropenyl, 2-Methylprop-2-enyl, 1-Methylprop-2-enyl, 2-Methylprop-1-enyl, (E)-1-Methylprop-1-enyl, (Z)-1-Methylprop-1-enyl, 3-Methylbuto-3-enyl, 2-Methylbuto-3-enyl, 1-Methylbuto-3-enyl, 3-Methylbuto These are to-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2-enyl, (Z)-1-methylbut-2-enyl, (E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl, (E)-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl, (E)-1-methylbut-1-enyl, (Z)-1-methylbut-1-enyl, 1,1-dimethylpropane-2-enyl, 1-ethylpropane-1-enyl, 1-propyl vinyl, and 1-isopropyl vinyl.

[0072] "C 2~10 An "alkynyl group" contains one or more triple bonds and has 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, for example, 2, 3, 4, 5, 6, 7, or 8 carbon atoms (i.e., "C 2~8 An alkynyl group) having 2, 3, 4, 5 or 6 carbon atoms (i.e., "C2~6 "Alkynyl group"), having 2 or 3 carbon atoms ("C 2~3 The term "alkynyl group" should preferably be understood to mean a linear or branched monovalent hydrocarbon group. Examples of the alkynyl group include ethynyl, propa-1-inyl, propa-2-inyl, buta-1-inyl, buta-2-inyl, buta-3-inyl, penta-1-inyl, penta-2-inyl, penta-3-inyl, penta-4-inyl, hexa-1-inyl, hexa-2-inyl, hexa-3-inyl, hexa-4-inyl, hexa-5-inyl, 1-methylpropa-2-inyl, 2-methylbuta-3-inyl, 1-methylbuta-3-inyl, 1-methylbuta-2-inyl, 3-methylbuta-1-inyl, 1-ethylpropa-2-inyl, 3-methylpenta-4-inyl, and 2-methyl These are penta-4-inyl, 1-methylpenta-4-inyl, 2-methylpenta-3-inyl, 1-methylpenta-3-inyl, 4-methylpenta-2-inyl, 1-methylpenta-2-inyl, 4-methylpenta-1-inyl, 3-methylpenta-1-inyl, 2-ethylbuta-3-inyl, 1-ethylbuta-3-inyl, 1-ethylbuta-2-inyl, 1-propylpropa-2-inyl, 1-isopropylpropa-2-inyl, 2,2-dimethylbuta-3-inyl, 1,1-dimethylbuta-3-inyl, 1,1-dimethylbuta-2-inyl, or 3,3-dimethylbuta-1-inyl. In particular, the alkynyl group is an ethynyl group, propa-1-inyl, or propa-2-inyl.

[0073] "C 3~10 The term "cycloalkyl group" should be understood to mean a saturated monovalent monocyclic, bicyclic (e.g., bridging ring, spiro ring) hydrocarbon ring or tricycloalkane having 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. 3~10The cycloalkyl group may be a monocyclic hydrocarbon group, such as a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a cyclooctyl group, a cyclononyl group, or a cyclodecyl group; or a bicyclic hydrocarbon group, such as a bornyl group, an indolyl group, a hexahydroindolyl group, a tetrahydronaphthyl group, a decahydronaphthyl group, a dicyclo[2.1.1]hexyl, a dicyclo[2.2.1]heptyl, a dicyclo[2.2.1]heptenyl, a 6,6-dimethyldicyclo[3.1.1]heptyl, a 2,6,6-trimethyldicyclo[3.1.1]heptyl, a dicyclo[2.2.2]octanyl, a 2,7-diazaspiro[3,5]nonanyl, a 2,6-diazaspiro[3,4]octanyl; or a tricyclic hydrocarbon group, such as an adamantyl group.

[0074] Unless otherwise defined, the term “3- to 6-membered heterocyclyl group” refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5-, or 6-membered monoring containing at least one heteroatom selected from O, S, and N, e.g., 1, 2, 3, 4, 5, or more, where N and S may be optionally oxidized to various oxidation states to form nitrogen oxides, -S(O)-, or -S(O)2-. The heterocyclyl group may include condensed or bridged rings and spirorings. In particular, the heterocyclyl group may include, but is not limited to, a four-membered ring such as an azetidinyl group or an oxetanyl group, a five-membered ring such as a tetrahydrofuranyl group, a dioxolyl group, a pyrrolidinyl group, an imidazolidinyl group, a pyrazolidinyl group, or a six-membered ring such as a tetrahydropyranyl group, a piperidinyl group, a morpholinyl group, a dithianyl group, a thiomorpholinyl group, a piperazinyl group, or a trithianyl group.

[0075] "C 6~14 The term "aryl group" refers to a monovalent aromatic or partially aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring having 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon atoms ("C 6~14"Aryl group"), especially a ring having 6 carbon atoms ("C6 aryl group"), for example a phenyl group or a biphenyl group, or a ring having 9 carbon atoms ("C9 aryl group"), for example an indanyl group or an indenyl group, or a ring having 10 carbon atoms ("C 10 "aryl group"), for example, a tetrahydronaphthyl group, a dihydronaphthyl group or a naphthyl group, or a ring having 13 carbon atoms ("C 13 "aryl group"), for example, a fluorenyl group, or a ring having 14 carbon atoms ("C" 14 It should be understood that the term "aryl group" preferably means an anthracenyl group, for example. 6-20 When an aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, the substitution position is not limited and may be, for example, ortho, para, or meta substitution.

[0076] The term “5-14 membered heteroaryl group” should be understood to include monovalent monocyclic, bicyclic, or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 5, 6, 9, or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3, heteroatoms independently selected from N, O, and S, each of which may be benzo-condensed. “Hyperaryl group” also refers to a group in which its heteroaromatic ring is condensed with one or more aryl groups, alicyclic groups, or heterocyclyl rings, where the basis or point of linkage lies in the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7- or 8-indolidinyl groups, 1-, 3-, 4-, 5-, 6- or 7-isoindolyl groups, 2-, 3-, 4-, 5-, 6- or 7-indolidinyl groups, 2-, 3-, 4-, 5-, 6- or 7-indazolyl groups, 2-, 4-, 5-, 6-, 7- or 8-prinyl groups, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-quinolidinyl groups, 2-, 3-, 4-, 5-, 6 -, 7- or 8-quinolinyl group, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolinyl group, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl group, 2-, 3-, 4-, 5- or 6-naphthilidinyl group, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl group, 3-, 4-, 5-, 6-, 7- or 8-synnolinyl group, 2-, 4-, 6- or 7-pteridinyl group, 1-, 2-,3-,4-,5-,6-,7- or 8-4aH carbazolyl group, 1-,2-,3-,4-,5-,6-,7- or 8-carbazolyl group, 1-,3-,4-,5-,6-,7-,8- or 9-carbolinyl group, 1-,2-,3-,4-,6-,7-,8-,9- or 10-phenantridinyl group, 1-,2-,3-,4-,5-,6-,7-,8- or 9-acridinyl group, 1-,2-,4-,5-,6- , 7-, 8- or 9-dinyl group, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthrolinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenadinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenoxadinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenadinyl group, 2-, 3-, 4-, 5-, 6- or 1-, 3-, 4-, 5-,6-,7-,8-,9- or 10-benzoisoquinolinyl group, 2-,3-,4- or thieno[2,3-b]furanyl group, 2-,3-,5-,6-,7-,8-,9-,10- or 11-7H-pyrazino[2,3-c]carbazolyl, 2-,3-,5-,6- or 7-2H-flo[3,2-b]-pyranyl, 2-,3-,4-,5-,7- or 8-5H-pyrido[2 ,3-d]-o-azinyl, 1-,3- or 5-1H-pyrazolo[4,3-d]-azolyl, 2-,4- or 54H-imidazo[4,5-d]thiazolyl, 3-,5- or 8-pyrazino[2,3-d]pyridazinyl, 2-,3-,5- or 6-imidazo[2,1-b]thiazolyl, 1-,3-,6-,7-,8- or 9-flo[3,4-c]sinnolinyl, 1-, 2-,3-,4-,5-,6-,8-,9-,10 or 11-4H-pyrido[2,3-c]carbazolyl, 2-,3-,6- or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-,4-,5-,6- or 7-benzoazolyl group, 2-,4-,5-,6- or 7-benzimidazolyl group, 2-,4-,4-,5-,6- or It contains a 7-benzothiazolyl group, a 1-,2-,4-,5-,6-,7-,8- or 9-benzoxapinyl group, a 2-,4-,5-,6-,7- or 8-benzoazinyl group, and a 1-,2-,3-,5-,6-,7-,8-,9-,10- or 11-4H-pyrrolo[1,2-b][2]benzoazepinyl. Typical condensed heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolinyl groups, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolinyl groups, 2-, 3-, 4-, 5-, 6- or 7-indolyl groups, 2-, 3-, 4-, 5-, 6- or 7-benzo[b]thienyl groups, 2-, 4-, 5-, 6- or 7-benzoazolyl groups, 2-, 4-, 5-, 6- or 7-benzimidazolyl groups, and 2-, 4-, 5-, 6- or 7-benzothiazolyl groups. When the 5- to 20-membered heteroaryl group is bonded to other groups to constitute the compound of the present invention, carbon atoms on the 5- to 20-membered heteroaryl ring may be bonded to other groups, or heteroatoms on the 5- to 20-membered heteroaryl ring may be bonded to other groups. The aforementioned 5-20 member heteroaryls, if substituted,It may be monosubstituted or polysubstituted. Furthermore, the substitution site is not limited; for example, hydrogen atoms bonded to carbon atoms on a heteroaryl ring may be substituted, or hydrogen atoms bonded to heteroatoms on a heteroaryl ring may be substituted.

[0077] The term "spiro ring" refers to a ring system in which two rings share a single ring-forming atom.

[0078] The term "condensed ring" refers to a ring system in which two rings share two ring-forming atoms.

[0079] The term "bridged ring" refers to a ring system in which two rings share three or more ring-forming atoms.

[0080] A wavy line intersecting a chemical bond. [ka] This symbol is used to indicate the linkage position between a group and an atom or group in the general formula structure.

[0081] Unless otherwise specified, heterocyclyl groups, heteroaryl groups, or heteroarylene groups include all possible isomeric forms thereof, e.g., their positional isomers. Thus, some exemplary and non-limiting examples may include forms in which they are substituted at one, two or more positions (if any) such as the 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-positions, or bonded to other groups, and include thienyl or thienylene groups containing pyridine-2-yl, pyridine-2-ylidene, pyridine-3-yl, pyridine-3-ylidene, pyridine-4-yl and pyridine-4-ylidene, thiophene-2-yl, thiophene-2-ylidene, thiophene-3-yl and thiophene-3-ylidene, pyrazole-1-yl, pyrazole-3-yl, pyrazole-4-yl and pyrazole-5-yl.

[0082] The term "alkyloxy group (alkoxy group)" refers to -O-(alkyl), where the definition of alkyl is as described above. Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, and butoxy groups. Alkoxy groups may be optionally substituted or unsubstituted. If substituted, the substituent is preferably one or more groups independently selected from alkyl groups, alkenyl groups, alkynyl groups, alkyloxy groups, alkylamino groups, halogens, mercapto groups, hydroxyl groups, nitro groups, cyano groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroaryl groups, cycloalkyloxy groups, or heterocycloalkyloxy groups.

[0083] The term "alkylamino group" refers to -NH- (alkyl group), and the definition of an alkyl group is as described above. Non-exclusive examples of alkylamino groups include methylamino group, ethylamino group, propylamino group, isopropylamino group, and butylamino group.

[0084] The term "(alkyl)2amino" refers to -N-(alkyl)2, where the definition of alkyl is as described above. Non-limiting examples of (alkyl)2amino include dimethylamino group, methylethylamino group, diethylamino group, dipropylamino group, methylpropylamino group, diisopropylamino group, dibutylamino group, etc.

[0085] A "haloalkyl group" refers to an alkyl group substituted with one or more halogens, where an alkyl group is defined as described above.

[0086] The term "antibody" refers to an immunoglobulin-derived molecule that can specifically bind to a target antigen, and the immunoglobulin-derived molecule binds to the target antigen via at least one antigen-binding site located in its variable region. When referring to the term "antibody," unless otherwise explicitly stated in the context, it includes not only complete antibodies but also antigen-binding fragments that can specifically bind to a target antigen. A "complete antibody" typically consists of two pairs of polypeptide chains (each pair having a light chain (LC) and a heavy chain (HC)). Antibody light chains can be classified into κ (kappa) and λ (lambda) light chains. Heavy chains can be classified into μ, δ, γ, α, or ε, and the antibody isotypes are defined as IgM, IgD, IgG, IgA, and IgE, respectively. Within the light and heavy chains, the variable and constant regions are connected by "J" regions of about 12 or more amino acids, and the heavy chain also contains "D" regions of about 3 or more amino acids. Each heavy chain consists of a variable heavy chain region (VH) and a constant heavy chain region (CH). The constant heavy chain region consists of three domains (CH1, CH2, CH3). Each light chain consists of a variable light chain region (VL) and a constant light chain region (CL). The constant light chain region consists of one domain CL. The constant domain does not directly participate in antibody binding to antigens, but exhibits various effector functions. For example, it can mediate the binding of immunoglobulins to various cells of the immune system (e.g., effector cells) and host tissues or factors containing the first component of the classical complement system (C1q). The VH and VL regions can also be subdivided into highly variable regions (called complementarity-determining regions (CDRs)) interspersed with relatively conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxyl terminus. The variable regions (VH and VL) of each heavy / light chain pair form antigen-binding sites.The assignment of amino acids in each region or domain may follow the definitions of Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk (1987) J. Mol. Biol. 196:901-917, or Chothia et al. (1989) Nature 342:878-883. As used herein, the terms “complementarity-determining region” or “CDR” refer to amino acid residues within the antibody variable region involved in antigen binding. The heavy chain and light chain variable regions each contain three CDRs, designated CDR1, CDR2, and CDR3. The precise boundaries of these CDRs can be defined according to various numbering systems known in the art. For example, CDRs can be defined according to the Kabat numbering system (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), the Chothia numbering system (Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883), or the IMGT numbering system (Lefranc et al., Dev. Comparat.Immunol. 27:55-77, 2003). For a given antibody, those skilled in the art will readily identify the CDR defined by each numbering system. Furthermore, the correspondences between various numbering systems are well known to those skilled in the art (see, for example, Lefranc et al., Dev. Comparat.Immunol. 27:55-77, 2003). In the present invention, the CDR contained in the antibody or its antigen-binding fragment can be determined according to various numbering systems known in the art.In some embodiments, the CDRs contained in the antibody or its antigen-binding fragment of the present invention are preferably determined according to the Kabat numbering system. As used herein, the terms “framework region” or “FR” residues refer to those amino acid residues in the antibody variable region other than the CDR residues defined above. The term “antibody” is not limited to any particular method of producing the antibody. For example, recombinant antibodies, monoclonal antibodies and polyclonal antibodies are included. The antibody may be an antibody of a different isotype, e.g., IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtype), IgA1, IgA2, IgD, IgE, or IgM antibody. As used herein, the term “antigen-binding fragment” of an antibody refers to a polypeptide comprising a fragment of a full-length antibody, which retains the ability to specifically bind to the same antigen to which the full-length antibody is bound, and / or competes with the full-length antibody for specific binding to the antigen, and is also called the “antigen-binding moiety.” Generally, see Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nd edition, Raven Press, NY (1989)), the full text of which is incorporated herein by reference for all purposes. Antigen-binding fragments of antibodies can be generated by recombinant DNA technology or by enzymatic or chemical cleavage of complete antibodies. Non-limiting examples of antigen-binding fragments include Fab, Fab', F(ab')2, Fd, Fv, Fv linked by disulfide bonds, scFv, di-scFv, diabody antibodies, single-domain antibodies, and such polypeptides, including at least a portion of antibodies sufficient to confer specific antigen-binding ability to the polypeptides. Engineered antibody variants are collectively described in Holliger et al., 2005; Nat Biotechnol, 23: 1126-1136.As used herein, the term "Fd" means an antibody fragment consisting of VH and CH1 domains, the term "Fab fragment" means an antibody fragment consisting of VL, VH, CL, and CH1 domains, the term "F(ab')2 fragment" means an antibody fragment containing two Fab fragments linked by disulfide crosslinks on a hinge region, and the term "Fab' fragment" refers to a fragment obtained after reductive bonding of the disulfide bonds of the two heavy chain fragments in the F(ab')2 fragment, consisting of complete light and heavy chain Fd fragments (consisting of VH and CH1 domains). As used herein, the term "Fv" means an antibody fragment consisting of the VL and VH domains of a single arm of the antibody. Fv fragments are generally considered to be the smallest antibody fragments capable of forming a complete antigen-binding site. Generally, six CDRs are considered to give an antibody its antigen-binding specificity. However, even a single variable region (e.g., an Fd fragment containing only three antigen-specific CDRs) can recognize and bind to an antigen, although its affinity may be lower than that of a complete binding site. As used herein, the term "Fc" refers to an antibody fragment formed by disulfide bonds between the second and third constant regions of the primary heavy chain of the antibody and the second and third constant regions of the secondary heavy chain. Antibody Fc fragments have a variety of different functions but are not involved in antigen binding. "Effector functions" mediated by the Fc domain include Fc receptor binding, Clq binding and complement-dependent cell-mediated cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, downregulation of cell surface receptors (e.g., B cell receptors), and B cell activation. The Fc domain may include a native Fc region or a mutant Fc region. A native Fc region contains an amino acid sequence that matches the amino acid sequence of Fc regions found in nature. For example, the native human Fc region includes the native human IgG1 Fc region, the native human IgG2 Fc region, the native human IgG3 Fc region, the native human IgG4 Fc region, and naturally occurring variants thereof. The mutant Fc region contains an amino acid sequence that differs from the amino acid sequence of the natural Fc region due to at least one amino acid modification.In some embodiments, the mutant Fc region may have modified effector functions (e.g., Fc receptor binding, antibody glycosylation, number of cysteine ​​residues, effector cell function, or complement function) compared to the native Fc region. As used herein, the term "scFv" refers to a single polypeptide chain comprising VL and VH domains, where the VL and VH are linked via a linker. Such scFv molecules may have the common structure NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. A suitable prior art linker consists of a repeating GGGGS amino acid sequence or a variant thereof. For example, a linker having the amino acid sequence (GGGGS)4 may be used, or a variant thereof may be used. In some cases, a disulfide bond may be present between the VH and VL of the scFv. In some embodiments of the present invention, scFv can form a di-scFv, which refers to an antibody formed by linking two or more single scFvs in series. In some embodiments of the present invention, an scFv can form (scFv)2, which refers to an antibody formed by the parallel linking of two or more single scFvs. As used herein, the term “biantibody” means that its VH and VL domains are expressed on a single polypeptide chain, but the use of a linker that is too short prevents pairing between the two domains on the same chain, thereby causing the domains to pair with complementary domains on another chain to produce two antigen-binding sites (see, for example, Holliger P. et al., Proc. Natl.Acad.Sci.USA 90:6444-6448 (1993), and Poljak RJ et al., Structure 2:1121-1123 (1994)). As used herein, the term “single-domain antibody (sdAb)” has the meaning commonly understood by those skilled in the art, and refers to an antibody fragment consisting of a single monomeric variable antibody domain (e.g., a single heavy chain variable region) that retains the ability to specifically bind to the same antigen to which a full-length antibody is bound.As used herein, the term “bispecific antibody” refers to an antibody that has binding specificity to two different antigens (or epitopes). The term “multispecific antibody” refers to an antibody that has binding specificity to at least two (e.g., three or four) different antigens (or epitopes). A bispecific or multispecific antibody comprises multiple antigen-binding domains that have binding specificity to different antigens (or epitopes), thereby enabling it to bind to at least two different binding sites and / or target molecules. Each antigen-binding domain in a bispecific or multispecific antibody can be independently selected from a full-length antibody (e.g., an IgG antibody) or its antigen-binding fragment (e.g., an Fv fragment, a Fab fragment, an F(ab')2 fragment, or scFv). In some cases, each antigen-binding domain is linked by a peptide linker. Each of the above antibody fragments retains the ability to specifically bind to the same antigen to which the full-length antibody is bound, and / or competes with the full-length antibody for specific binding to the antigen. An antigen-binding fragment of an antibody (e.g., the antibody fragment mentioned above) is obtained from a predetermined antibody (e.g., the antibody provided by the present invention) by conventional techniques known to those skilled in the art (e.g., recombinant DNA techniques or enzymatic or chemical cleavage methods), and then complete antibody is obtained. Antigen-binding fragments of antibodies can be specifically screened using the same methods as those used in the human body. As used herein, the term "humanized antibody" refers to a genetically modified non-human antibody whose amino acid sequence has been modified to improve sequence homology with that of a human antibody. Generally, all or part of the CDR region of a humanized antibody is derived from a non-human antibody (donor antibody), and all or part of the non-CDR region (e.g., the variable region FR and / or constant region) is derived from a human immunoglobulin (receptor antibody). In some embodiments, the CDR region of a humanized antibody is derived from a non-human antibody (donor antibody), and all or part of the non-CDR region (e.g., the variable region FR and / or constant region) is derived from a human immunoglobulin (receptor antibody). Humanized antibodies generally retain the expected properties of the donor antibody, including but not limited to antigen specificity, affinity, and reactivity. In this application, the donor antibody may be a mouse antibody having the expected properties (e.g., antigen specificity, affinity, reactivity, etc.). To produce a humanized antibody, the CDR region of a donor antibody can be inserted into a human framework sequence using methods known in the art. In some cases, the human framework sequence may include amino acid mutations substituted with corresponding non-human residues. The humanized antibody may further include residues not found in either the initial donor antibody variable region (e.g., the light chain variable region or the heavy chain variable region) or the human framework sequence in order to further improve or optimize the function of the humanized antibody. As used herein, the term “chimeric antibody” refers to an antibody in which part of its light chain and / or heavy chain originates from one antibody (which may originate from a particular species or belong to a particular antibody class or subclass), and another part of its light chain and / or heavy chain originates from another antibody (which may originate from the same or different species or belong to the same or different antibody class or subclass), but in any case retains binding activity to the target antigen. In some embodiments, the term "chimeric antibody" may include antibodies in which the heavy chain variable region and light chain variable region of the antibody originate from a first antibody, but the heavy chain constant region and light chain constant region of the antibody originate from a second antibody.

[0087] As those skilled in the art will understand, the compounds represented by formula (I) may exist in various pharmaceutically acceptable salt forms. If these compounds have a basic center, they can form acid-added salts; if they have an acidic center, they can form base-addition salts; and if they contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they can also form intramolecular salts.

[0088] The compounds of the present invention may exist in the form of solvates (e.g., hydrates), wherein the compounds of the present invention include a polar solvent, particularly water, methanol, or ethanol, as a component of the compound crystal lattice. The amount of the polar solvent, particularly water, may be in stoichiometric or non-stoichiometric ratios.

[0089] Depending on their molecular structure, the compounds of the present invention may be chiral, and therefore various enantiomer forms may exist. Accordingly, these compounds can exist in racemic or optically active forms. The compounds of the present invention cover isomers or mixtures thereof, where each chiral carbon is in an R or S configuration, and racemates. The compounds of the present invention or their intermediates can be separated into enantiomer compounds by chemical or physical methods known to those skilled in the art, or can be used in synthesis in such forms. In the case of racemized amines, diastereomers were prepared from the mixture by reaction with an optically active resolution reagent. Examples of suitable resolution reagents include optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid in R and S configurations, suitable N-protected amino acids (e.g., N-benzoylproline or N-benzenesulfonylproline), or various optically active camphorsulfonic acids. Optically active resolution reagents (e.g., dinitrobenzoylphenylglycine, cellulose triacetate, or other carbohydrate derivatives or chiral derivatized methacrylate polymers immobilized on silica gel) can be used to advantageously perform enantiomeric resolution in chromatography. Suitable eluents for this purpose are water or alcohol-containing solvent mixtures, such as hexane / isopropanol / acetonitrile.

[0090] The corresponding stable isomers can be separated by known methods, such as extraction, filtration, or column chromatography.

[0091] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, and most preferably humans.

[0092] The term “therapeutic dose” refers to the amount of an active compound or drug that researchers, veterinarians, physicians or other clinicians seek from a tissue, system, animal, individual or human to produce a biological or medical response, and it includes one or more of the following: (1) prevention of disease: e.g., preventing disease, disorder or disorder in an individual that is susceptible to infection but has not yet experienced or developed the pathology or symptoms of the disease; (2) inhibition of disease: e.g., inhibiting disease, disorder or disorder in an individual that has experienced or developed the pathology or symptoms of disease, disorder or disorder (i.e., preventing further progression of the pathology and / or symptoms); (3) remission of disease: e.g., relieving disease, disorder or disorder in an individual that has experienced or developed the pathology or symptoms of disease, disorder or disorder (i.e., reversing the pathology and / or symptoms). [Brief explanation of the drawing]

[0093] [Figure 1] This is a schematic diagram of ADC-5 free toxin release in mouse, rat, human, and monkey plasma. [Figure 2] This is a schematic diagram of ADC-10 free toxin release in mouse, rat, human, and monkey plasma. [Figure 3] This is a schematic diagram of the release of DS8201 toxins in mouse, rat, human, and monkey plasma. [Figure 4] This is a schematic diagram of the release of free toxins by ADC-5, ADC-10, ADC-14, and ADC-15 in mouse plasma. [Figure 5] This is a schematic diagram of the release of free toxins by ADC-5, ADC-10, ADC-14, and ADC-15 in human plasma. [Figure 6] This is a schematic diagram of the drug efficacy evaluation in NCI-N87 tumor-carrying mice. [Figure 7] This is a schematic diagram of the drug efficacy evaluation in JIMT-1 tumor-carrying mice. [Modes for carrying out the invention]

[0094] The following provides a more detailed explanation of the technical aspects of the present invention with specific examples. It should be understood that the following examples are merely illustrative and interpretive of the present invention and should not be interpreted as limiting the scope of protection of the present invention. Any technology realized based on the above-described aspects of the present invention falls within the intended scope of protection of the present invention.

[0095] Unless otherwise specified, the raw materials and reagents used in the following examples are either commercially available or can be manufactured by known methods.

[0096] The structure of the compound is determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The NMR shift (δ) is 10 -6 The values ​​are expressed in units of ppm. A Bruker AVANCE-400 nuclear magnetic resonance spectrometer is used for NMR measurements, and the measurement solvents are deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard.

[0097] For MS measurements, an Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS liquid chromatograph mass spectrometer (Manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS) was used. Other instruments used included the waters ACQuity UPLC-QD / SQD (Manufacturer: waters, MS model: waters ACQuity Qda Detector / waters SQ Detector) and the THERMO Ultimate 3000-Q Exactive (Manufacturer: THERMO, MS model: THERMO Q Exactive).

[0098] High-performance liquid chromatography (HPLC) analysis was performed using an Agilent 1260II HPLC and a Waters Acquity UPLC H-Class high-performance liquid chromatograph.

[0099] Chiral HPLC analysis was performed using a Waters Acquity UPCC high-performance liquid chromatograph.

[0100] High-performance liquid preparative chromatography was performed using Waters MS-triggered Prep-LC with SQD2 detector, Waters MS triggered Prep-LC with Acquity QDA detector, Waters MS-triggered Prep-LC with QDA detector, and GILSON Prep LC with UV detector.

[0101] For the CombiFlash flash chromatograph, we used the Combiflash Rf200 (TELEDYNE ISCO).

[0102] For thin-layer chromatography, Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates are used. The size of the silica gel plates used for thin-layer chromatography (TLC) is 0.15 mm to 0.2 mm, and the size of the separated and purified products by thin-layer chromatography is 0.4 mm to 0.5 mm.

[0103] Silica gel column chromatography typically uses 200-300 mesh silica gel as the support material.

[0104] Kinase mean inhibition rate and IC 50 The values ​​were measured using a NovoStar microblade reader (BMG GmbH, Germany).

[0105] The known starting materials of the present invention may be synthesized using or in accordance with methods known in the art, or may be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., and Dalui Chemicals.

[0106] Unless otherwise stated in the examples, the reactions may be carried out under an argon or nitrogen atmosphere.

[0107] An argon or nitrogen atmosphere refers to a reaction flask being connected to an argon or nitrogen balloon with a volume of approximately 1 liter.

[0108] A hydrogen atmosphere refers to a reaction flask being connected to a hydrogen balloon with a volume of approximately 1 liter.

[0109] The pressurized hydrogenation reaction was carried out using a Parr 3916EKX type hydrogenator and a QL-500 type hydrogen gas generator or an HC2-SS type hydrogenator.

[0110] The hydrogenation reaction was typically carried out by evacuating the system, filling it with hydrogen gas, and repeating the operation three times.

[0111] The microwave reaction was performed using a CEM Discover-S 908860 microwave reactor.

[0112] Unless otherwise specified in the examples, "solution" refers to an aqueous solution.

[0113] Unless otherwise specified in the examples, the reaction temperature is room temperature, between 20°C and 30°C.

[0114] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system for column chromatography used for compound purification, and the developing solvent system for thin-layer chromatography included A: dichloromethane / methanol system and B: n-hexane / ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compound, and small amounts of basic or acidic reagents such as triethylamine and acetic acid were optionally added to adjust the ratio.

[0115] ADC preparative analyzer: Benchtop centrifuges used were Eppendoff 5810R, 5430R, and 5418R.

[0116] A METTLER TOLEDO Seven Ecellence pH meter and a METTLER TOLEDO ME1002E electronic balance were used. An Eppendorf 5382KN644547 thermomixer was used. An IKA LoopSTER inversion mixer was used. A Thermo Scientific 2000-C nanodrop spectrophotometer was used. A BioTek EPOCH2 microblade reader was used. LC-MS was performed using a 6224 TOF and a 6530 LC / Q-TOF, and HPLC was performed using an Agilent Technologies 1260 Infinity II.

[0117] Example 1 (R)-N-((1S,9S)-9-ethyl-5-fluoro-4,9-dihydroxy-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)-3-hydroxybutanamide (R)-N-((1R,9S)-9-ethyl-5-fluoro-4,9-dihydroxy-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)-3-hydroxybutanamide [ka]

[0118] Step 1: N-(3-bromo-5-fluoro-4-methoxyphenyl)acetamide 1b 3-Bromo-5-fluoro-4-methoxyaniline 1a (20 g, 90.8 mmol) was dissolved in dichloromethane (40 mL), cooled to 0°C, and then acetyl chloride (14.3 g, 181.6 mmol) and triethylamine (27.6 mg, 272.4 mmol) were slowly added. The reaction mixture was stirred at 0°C for 0.5 hours. After the reaction was complete, water (30 mL) was added to the reaction mixture, and it was extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 1b (20 g, yield: 84%). MS m / z (ESI): 262.1 (M+H) + .

[0119] Step 2 (E)-4-(5-acetamido-3-fluoro-2-methoxyphenyl)buta-3-enoic acid 1c Compound 1b (20 g, 76.3 mmol) was dissolved in dioxane (30 mL) and water (10 mL), and buta-3-enoic acid (7.23 g, 83.9 mmol), palladium acetate (1.71 g, 7.6 mmol), tris(o-methylphenyl)phosphine (4.64 g, 15.2 mmol), and N,N-diisopropylethylamine (30 mg, 229 mmol) were added. The reaction mixture was stirred at 100 °C for 16 hours. After the reaction was complete, the reaction mixture was filtered, and the resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 1c (20 g, yield: 87%). MS m / z (ESI): 268.1 (M+H) + .

[0120] Step 3: 4-(5-acetamido-3-fluoro-2-methoxyphenyl)butyrate 1d Compound 1c (20 g, 74.8 mmol) was dissolved in tetrahydrofuran (50 mL), and 10% Pd / C (0.8 g, 7.4 mmol) was added. The reaction mixture was stirred at room temperature under a hydrogen gas atmosphere for 2 hours. After the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The resulting residue was purified using silica gel column chromatography with eluent system A to obtain the title compound 1d (20 g, yield: 99%). MS m / z (ESI): 270.1 (M+1) + .

[0121] Step 4 N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 1e Compound 1d (20 g, 74.4 mmol) was dissolved in trifluoroacetic acid (60 mL), cooled to 0°C, and trifluoroanhydride (31.24 g, 148.8 mmol) was slowly added. The reaction mixture was stirred at room temperature for 7 hours. After the reaction was complete, the reaction mixture was slowly poured into water (50 mL), extracted with ethyl acetate (50 mL x 3), the organic phases were combined, washed with saturated sodium bicarbonate aqueous solution until neutral, further washed with saturated brine, dried, and concentrated. The resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 1e (9.2 g, yield: 46%). MS m / z (ESI): 252.1 (M+1) + .

[0122] Step 5 (Z)-N-(3-fluoro-7-(hydroxyimino)-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 1f Potassium tert-butoxide (9.83 g, 87.5 mmol) was dissolved in tetrahydrofuran (40 mL) and tert-butanol (10 mL), and after cooling to 0°C, N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 1e (10 g, 39.8 mmol) was dissolved in tetrahydrofuran (10 mL) and slowly added to the reaction mixture. After 10 minutes, isoamyl nitrite (7.46 g, 63.6 mmol) was added. The reaction mixture was stirred at 0°C for 1 hour. After the reaction was complete, saturated ammonium chloride solution was added to the reaction mixture to quench it, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 1f (6 g, yield: 51%). MS m / z (ESI): 281.1 (M+1) + .

[0123] Step 6: N-(7-amino-3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 1g (Z)-N-(3-fluoro-7-(hydroxyimino)-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 1f (6g, 21.4 mmol) was dissolved in dioxane (60 mL) and 2N hydrochloric acid solution (20 mL), and 10% Pd / C (1.13 g, 10.7 mmol) was added. The reaction mixture was stirred at room temperature under a hydrogen gas atmosphere for 5 hours. After the reaction was complete, the reaction mixture was filtered and concentrated to obtain 1 g (5 g) of crude product, which was used directly in the next step without purification. MS m / z (ESI): 267.1 (M+1) + .

[0124] Step 7 (9H-Fluoren-9-yl)methyl(8-acetamido-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate 1h 1 g (5 g, 18.8 mmol) of the compound obtained in the previous step was dissolved in dioxane (50 mL), and the pH was adjusted to 7-8 with sodium bicarbonate. Then, 6.36 g (18.8 mmol) of 9-fluorenylmethyl-N-succinimidyl carbonate was slowly added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was slowly poured into water (40 mL), extracted with ethyl acetate (40 mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated.

[0125] The resulting residue was purified using silica gel column chromatography with eluent system B to obtain 1h (5.3g, yield: 54%) of the title compound. MS m / z (ESI): 489.2 (M+1) + .

[0126] Step 8 (9H-Fluoren-9-yl)methyl(8-amino-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate 1i Compound 1h (5g, 10.25 mmol) was dissolved in dioxane (50 mL) and 12N hydrochloric acid (10 mL). The reaction mixture was stirred at 60°C for 2 hours. After the reaction was complete, the reaction mixture was slowly poured into water (50 mL), extracted with ethyl acetate (40 mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 1i (3.75 g, yield: 78%). MS m / z (ESI): 447.2 (M+1) + .

[0127] Step 9 (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]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)carbamate 1k Compound 1i (3 g, 6.6 mmol) was dissolved in toluene (30 mL), and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-indridine-3,6,10(4H)-trione 1j (2.61 g, 9.9 mmol) and p-toluenesulfonic acid (2.52 g, 13.2 mmol) were added. The reaction mixture was stirred at 110 °C for 5 hours. After the reaction was complete, the mixture was extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 1k (1.8 g, yield: 36%). MS m / z (ESI): 674.2 (M+1) + .

[0128] Step 10 (9H-Fluoren-9-yl)methyl((9S)-9-ethyl-5-fluoro-4,9-dihydroxy-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 1l Compound 1k (1.8g, 2.7 mmol) was dissolved in 40% hydrobromic acid (40mL). The reaction mixture was stirred at 100°C for 2 hours. After the reaction was complete, the reaction mixture was extracted with ethyl acetate (50mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting residue was purified using silica gel column chromatography with eluent system B to obtain 1 liter (1.08g, yield: 60%) of the title compound. MS m / z (ESI): 660.2 (M+1) + .

[0129] Step 11 (9S)-1-amino-9-ethyl-5-fluoro-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-10,13-dione 1m 1 liter (500 mg, 0.7 mmol) of the compound was dissolved in 5 mL of N,N-dimethylformamide, and diethylamine (166 mg, 2.3 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the diethylamine in the reaction mixture was evaporated by rotation using an oil pump, and 1 ml of the crude product obtained by rotational evaporation was beaten with ethyl acetate. The resulting solid was directly added to the reaction in the next step. MS m / z (ESI): 438.1 (M+1) + .

[0130] Step 12 (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((R)-3-hydroxybutyrylamino)-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-4-yl-(R)-3-hydroxybutyrate 1n Compound 1m (100 mg, 0.2 mmol) was dissolved in N,N-dimethylformamide (3 mL), and (R)-3-hydroxybutyric acid (36 mg, 0.34 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (174 mg, 0.46 mmol), and N,N-diisopropylethylamine (88 mg, 0.68 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was extracted with ethyl acetate (50 mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 1n (90 mg, yield: 64%). MS m / z (ESI): 610.2 (M+1) + .

[0131] Step 13 (R)-N-((1S,9S)-9-ethyl-5-fluoro-4,9-dihydroxy-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)-3-hydroxybutanamide (R)-N-((1R,9S)-9-ethyl-5-fluoro-4,9-dihydroxy-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)-3-hydroxybutanamide Compound 1n (90 mg, 0.15 mmol) was dissolved in methanol (5 mL), and an aqueous solution of 1 M lithium hydroxide (2 mL) was added at room temperature. The reaction was stirred at room temperature for 15 minutes. After the reaction was complete, methanol was evaporated by rotation, and the remaining aqueous phase was freeze-dried to obtain the crude product. The crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge C18 150*19 mm, 5 μm; mobile phase 1: water (containing 0.1% TFA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 23%~33%, flow rate: 20 mL / min) to obtain compound 1-1 (35 mg, yield: 30%) and compound 1-2 (28 mg, yield: 24%).

[0132] Single-stereoconfiguration compound 1-1 (short retention time): MS m / z (ESI): 524.2 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 10.42 (s, 1H), 8.39 (d, J = 8.8 Hz, 1H), 7.84 (d, J = 11.6 Hz, 1H), 7.26 (s, 1H), 6.50 (s, 1H), 5.57 - 5.49 (m, 1H), 5.41 (s, 2H), 5.27 - 5.11 (m, 2H), 4.04 (dd, J = 13.2, 6.0 Hz, 1H), 3.18 - 3.04 (m, 2H), 2.36 - 2.17 (m, 2H), 2.15 - 1.96 (m, 2H), 1.92 - 1.77 (m, 2H), 1.08 (d, J = 6.4 Hz, 3H), 0.93 - 0.81 (m, 3H).

[0133] Single stereochemical compound 1-2 (long retention time): MS m / z (ESI): 524.2 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 10.42 (s, 1H), 8.42 (d, J = 8.8 Hz, 1H), 7.84 (d, J = 11.6 Hz, 1H), 7.26 (s, 1H), 5.59 - 5.51 (m, 1H), 5.42 (s, 2H), 5.29 - 5.14 (m, 2H), 4.04 (dd, J = 13.2, 6.4 Hz, 1H), 3.17 (dd, J = 12.0, 4.8 Hz, 1H), 3.10 - 2.92 (m, 1H), 2.28 (dd, J = 13.6, 7.2 Hz, 1H), 2.18 (dd, J = 13.6, 5.6 Hz, 1H), 2.13 - 1.95 (m, 2H), 1.93 - 1.78 (m, 2H), 1.08 (d, J = 6.0 Hz, 3H), 0.91 - 0.82 (m, 3H).

[0134] Example 2 (R)-N-((1S,9S)-9-ethyl-4,9-dihydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indridine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1R,9S)-9-ethyl-4,9-dihydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indridine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide [ka]

[0135] Using the synthesis route of Example 1, the starting material in Step 1 was replaced with 3-bromo-4-methoxyaniline 2a (25 g, 0.12 mol), and the product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge C18 150*19 mm, 5 μm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 23%~33%, flow rate: 20 mL / min) to obtain the title products 2-1 (19 mg, yield: 28%) and 2-2 (30 mg, yield: 44%).

[0136] Single-stereoconfiguration compound 2-1 (short retention time): MS m / z (ESI): 506 (M+1) + . 1H NMR (400MHz, DMSO-d6) δ 10.21 (s, 1H), 8.39- 8.37 (m, 1H), 7.92 - 7.90 (m, 1H), 7.51 - 7.50 (m, 1H), 7.24 (s, 1H), 6.49 (s, 1H), 5.57 - 5.48 (m, 1H), 5.41 (s, 2H), 5.27 - 5.11 (m, 2H), 4.65 (s, 1H), 4.09 - 3.98 (m, 1H), 3.09 - 2.96 (m, 2H), 2.25 - 2.23 (m, 2H), 2.05 - 2.00 (m, 2H), 1.95 - 1.77 (m, 2H), 1.23 (s, 2H), 1.08 - 1.07 (m, 3H), 0.86 - 0.85 (m, 3H).

[0137] Single stereo compound 2-2 (long retention time): MS m / z (ESI): 506 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 10.22 (s, 1H), 8.42 - 8.40 (d, J = 8.4 Hz, 1H), 7.93 - 7.90 (d, J =8.4 Hz, 1H), 7.51 - 7.50 (m, 1H), 7.25 (s, 1H), 5.58 - 5.46 (m,1H), 5.42 (s, 2H), 5.21 - 5.20 (m, 2H), 4.04 - 4.01 (m, 1H), 2.96 - 2.94 (m, 2H), 2.28 (s, 2H), 2.10 - 1.97 (m, 2H), 1.86 (s, 2H), 1.23 (s, 2H), 1.08 - 1.06 (m, 3H), 0.87 - 0.85 (m, 3H).

[0138] Example 3 (R)-N-((1S,9S)-4-bromo-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]indridine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1R,9S)-4-bromo-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]indridine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide [ka]

[0139] Step 1: 4-Bromo-3-Fluoro-5-iodoaniline 3b 3-Fluoro-5-iodoaniline 3a (50 g, 210.95 mmol) was dissolved in DMF (250 mL), and NBS (41.30 g, 232.04 mmol) was slowly added under ice bath conditions. The reaction was stirred at room temperature for 16 hours. After the reaction was complete, water was added, the system was extracted with dichloromethane, the organic phase was washed with saturated brine, the organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain crude product 3b (66 g). MS m / z (ESI): 315.9, 317.9 (M+1) + . 1 H NMR (400MHz, DMSO-d6) δ 7.01 (d, 1H), 6.47 (dd, 1H), 5.74 (s, 2H).

[0140] Step 2 N-(4-bromo-3-fluoro-5-iodophenyl)acetamide 3c Compound 3b (66 g, 208.92 mmol) was dissolved in dichloromethane (660 mL), and triethylamine (42.28 g, 417.84 mmol) was added. After cooling the system to 0°C, acetyl chloride (19.68 g, 250.70 mmol) was slowly added dropwise. After the addition was complete, the system was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product, which was then dissolved again in ethyl acetate. The pH of the system was adjusted to 2-3 with dilute hydrochloric acid, the system was extracted with ethyl acetate, washed with saturated brine, the organic phase was collected, and dried over anhydrous sodium sulfate. The solvent was evaporated by rotation to obtain the crude product, which was beaten with a mixed solvent of dichloromethane / methanol (10:1) to obtain the title compound 3c (60 g, yield: 80%). MS m / z (ESI): 357.9, 359.9 (M+1) + . 1 H NMR (400MHz, DMSO-d6) δ 10.27 (s, 1H), 7.96 (s, 1H), 7.66 (dd, 1H), 2.06 (s, 3H).

[0141] Step 3 (E)-4-(5-acetamido-2-bromo-3-fluorophenyl)buta-3-enoic acid 3d Compound 3c (20 g, 55.87 mmol) was dissolved in a mixed solvent of dioxane (200 mL) and water (40 mL), and buta-3-enoic acid (4.81 g, 55.87 mmol), DIPEA (14.45 g, 111.74 mmol), palladium acetate (630 mg, 2.79 mmol), and tris(o-methylphenyl)phosphine (1.7 g, 5.59 mmol) were added. The reaction was stirred at 100 °C for 16 hours under nitrogen gas protection. After the reaction was complete, water and dichloromethane were added, the system was washed 3 to 5 times with saturated sodium bicarbonate solution, and the aqueous phase was collected. The pH of the aqueous phase was adjusted to 2 to 3 with hydrochloric acid, and the organic phase was extracted 5 to 7 times with ethyl acetate to collect the organic phase, which was dried over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain the crude product compound 3d (17 g). MS m / z (ESI): 316.0, 317.9 (M+1) + . Step 4: 4-(5-acetamido-2-bromo-3-fluorophenyl)butyrate 3e

[0142] Compound 3d (8 g, 25.3 mmol) was dissolved in methanol (80 mL), and platinum-carbon catalyst (800 mg) was added. The reaction was stirred at room temperature for 2 hours under a hydrogen gas environment. After the reaction was complete, the mixture was filtered, the filtrate was collected, and the solvent was evaporated by rotation to obtain crude product 3e (8 g). The crude product was used directly in the next step without purification. MS m / z (ESI): 318.0, 320.0 (M+1) + . 1 H NMR (400MHz, DMSO-d6) δ 12.14 (s, 1H), 10.24 (s, 1H), 7.63 (dd, 1H), 7.25 (s, 1H), 2.75-2.67 (m, 2H), 2.29 (t, 2H), 2.05 (s, 3H), 1.79 (dd, 2H).

[0143] Step 5 N-(4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 3f Compound 3e (8 g, 25.1 mmol) was dissolved in trifluoroacetic acid (80 mL), and trifluoroanhydride (15.82 g, 75.3 mmol) was slowly added under ice bath conditions. The reaction was stirred at 0°C for 4 hours. After the reaction was complete, water was added under ice bath conditions, and the pH of the system was adjusted to 9-10 with a 15% sodium hydroxide solution. The system was then extracted with dichloromethane, the organic phase was washed with saturated brine, and the organic phase was collected. The system was dried over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain compound 3f (4.8 g, yield: 53%). MS m / z (ESI): 329.0, 331.0 (M+1) + .

[0144] Step 6 (Z) -N-(4-bromo-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 3g Potassium tert-butoxide (1.62 g, 14.4 mmol) was dissolved in a mixed solvent of tetrahydrofuran (80 mL) and tert-butanol (20 mL), and compound 3f (2.15 g, 7.2 mmol) in a tetrahydrofuran solution (20 mL) was slowly added under ice bath. The reaction was stirred at 0°C for 10 minutes, after which isoamyl nitrite (1.27 g, 10.8 mmol) was added, and the reaction was continued with stirring at 0°C for 50 minutes. After the reaction was complete, dilute hydrochloric acid was added to adjust the pH of the system to 4-5, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, collected, dried over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain the crude product. The crude product was beaten with methyl tert-butyl ether to obtain 3 g of the title compound (1.1 g, yield: 46%). MS m / z (ESI): 329.0, 331.0 (M+1) + . 1 H NMR (400MHz, CD3OD) δ 8.52 (d, 1H), 3.21 (dd, 2H), 3.07 (dd, 2H), 2.24 (s, 3H).

[0145] Step 7 N-(7-amino-4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 3h Three g (500 mg, 1.52 mmol) of the compound was dissolved in 10 mL of dioxane, and 1 mL of 1 M hydrochloric acid and 100 mg of platinum-carbon catalyst were added. The reaction was stirred at room temperature for 4 hours under the influence of hydrogen gas. After the reaction was complete, the mixture was filtered, the filtrate was collected and concentrated to obtain 3 h (500 mg) of crude product, which was used directly in the next step of the reaction. MS m / z (ESI): 315.1, 317.1 (M+1) + .

[0146] Step 8 (9H-Fluoren-9-yl)methyl(8-acetamido-5-bromo-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate 3i Compound 3h (500 mg) was dissolved in dioxane (10 mL), and the pH of the filtrate from step 7 was adjusted to 8-9 with saturated sodium carbonate solution. Then, fluorenyl methoxycarbonyl chloride (432 mg, 1.67 mmol) was added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the system was extracted with ethyl acetate, the organic phase was washed with saturated brine, the organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain the crude product. The resulting residue was purified by eluate system B using silica gel column chromatography to obtain the title compound 3i (300 mg, yield: 37%). MS m / z (ESI): 537.0, 539.0 (M+1) + .

[0147] Step 9 (9H-Fluoren-9-yl)methyl(8-amino-5-bromo-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate 3j Compound 3i (700 mg, 1.30 mmol) was dissolved in methanol (10 mL), and concentrated hydrochloric acid (12 mol / L, 2 mL) was added. The reaction was stirred at 60°C for 1 hour. After the reaction was complete, the resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 3j (500 mg, yield: 77%).

[0148] Step 10 ((9H-Fluoren-9-yl)methyl((9S)-4-bromo-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 3k Compound 3j (200 mg, 0.40 mmol) was dissolved in toluene (5 mL), and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f]indolidine-3,6,10(4H)-trione 1j (117 mg, 0.44 mmol) and p-toluenesulfonic acid monohydrate (77 mg, 0.40 mmol) were added. The reaction was stirred at 120 °C for 2 hours. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product, and the resulting residue was purified by eluate system B using silica gel column chromatography to obtain the title compound 3k (250 mg, yield: 86%). MS m / z (ESI): 722.0, 724.0 (M+1) + . 1 H NMR (400MHz, CDCl3) δ 8.19 - 7.31 (m, 9H), 5.68 (dd, 1H), 5.33 - 5.18 (m, 2H), 4.66 (s, 2H), 4.34 (d, 1H), 4.21 - 4.04 (m, 2H), 3.26 (s, 1H), 3.00 - 2.94 (m, 1H), 2.04 (s, 2H), 1.81 (s, 2H), 1.70 - 1.50 (m, 2H), 1.27 (dd, 3H).

[0149] Step 11 (9S)-1-amino-4-bromo-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-10,13-dione 3l Compound 3k (200 mg, 0.28 mmol) was dissolved in DMF (1 mL), and diethylamine (0.1 mL) was added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the resulting residue was purified using silica gel column chromatography with eluent system A to obtain 3 l (100 mg, yield: 72%) of the title compound. MS m / z (ESI): 500.0, 502.1 (M+1) + .

[0150] Step 12 (R)-N-((1S,9S)-4-bromo-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]indridine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1R,9S)-4-bromo-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]indridine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide 3 liters (100 mg, 0.20 mmol) of the compound were dissolved in 2 mL of DMF, and (R)-3-hydroxybutyric acid (25 mg, 0.24 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (114 mg, 0.30 mmol), and N,N-diisopropylethylamine (52 mg, 0.40 mmol) were added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the compounds were purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2*250 mm, 10 μm; mobile phase 1: water (0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 38%~48%, flow rate: 25 mL / min) to obtain compound 3-1 (1.62 mg, yield: 6%) and compound 3-2 (1.32 mg, yield: 7.4%).

[0151] Single-stereoconfiguration compound 3-1 (short retention time): MS m / z (ESI): 586.0, 588.0 (M+1) + . 1H NMR (400 MHz, CD3OD) δ 8.10 (d, J = 10.0 Hz, 1H), 7.90 (s, 1H), 5.96 - 5.88 (m, 1H), 5.83 (d, J = 16.4 Hz, 1H), 5.71 - 5.48 (m, 4H), 4.52 - 4.46 (m, 1H), 2.71 - 2.65 (m, 2H), 2.59 - 2.52 (m, 2H), 2.23 - 2.18 (m, 3H), 1.49 (d, J = 6.4 Hz, 3H), 1.25 (t, J = 7.2 Hz, 3H).

[0152] Single-stereoconfiguration compounds 3-2 (long retention time): MS m / z (ESI): 586.0, 588.0 (M+1) + . 1 H NMR (400 MHz, CD3OD) δ 7.79 (d, J = 9.6 Hz, 1H), 7.59 (s, 1H), 5.67 - 5.59 (m, 1H), 5.52 (d, J = 16.4 Hz, 1H), 5.41 - 5.33 (m, 2H), 5.32 - 5.23 (m, 2H), 4.24 - 4.18 (m, 1H), 2.34 (d, J = 6.8 Hz, 2H), 2.28 - 2.21 (m, 2H), 1.93 - 1.86 (m, 3H), 1.17 (d, J = 6.4 Hz, 3H), 0.94 (t, J = 7.2 Hz, 3H).

[0153] Example 4 (R)-N-((1S,9S)-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)-3-hydroxybutanamide (R)-N-((1R,9S)-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)-3-hydroxybutanamide [ka]

[0154] Using the synthesis route of Example 3, 3-fluoro-5-iodoaniline (25 g, 105.5 mmol) was used as the starting material and purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2*250 mm, 10 μm; mobile phase 1: water (0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 38%~48%, flow rate: 25 mL / min) to obtain the title products 4-1 (2.5 mg, yield: 12%) and 4-2 (3.1 mg, yield: 15%).

[0155] Single-stereoconfiguration compounds 4-1 (short retention time): MS m / z (ESI): 508.1 (M+1) + . 1 H NMR (400 MHz, CD3OD) δ 7.65 (d, J = 10.2 Hz, 2H), 7.37 (d, J = 8.8 Hz, 1H), 5.66 (s, 1H), 5.56 (d, J = 16.4 Hz, 1H), 5.40 - 5.20 (m, 4H), 4.28 - 4.21 (m, 1H), 2.47 - 2.39 (m, 2H), 2.27 (s, 2H), 1.98 - 1.90 (m, 2H), 1.24 (d, J = 6.4 Hz, 4H), 0.99 (t, J = 7.2 Hz, 3H).

[0156] Single-stereoconfiguration compounds 4-2 (long retention time): MS m / z (ESI): 508.1 (M+1) + . 1 H NMR (400 MHz, CD3OD) δ 7.57 (d, J = 6.2 Hz, 2H), 7.30 (d, J = 8.0 Hz, 1H), 5.60 (d, J = 6.0 Hz, 1H), 5.48 (d, J = 16.4 Hz, 1H), 5.36 - 5.15 (m, 5H), 4.22 - 4.15 (m, 1H), 2.36 - 2.30 (m, 2H), 2.18 (t, J = 6.4 Hz, 3H), 1.90 - 1.83 (m, 2H), 1.15 (d, J = 6.4 Hz, 3H), 0.91 (t, J = 7.2 Hz, 3H).

[0157] Example 5 (R)-N-((1S,9S)-4-(difluoromethoxy)-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)-3-hydroxybutanamide (R)-N-((1R,9S)-4-(difluoromethoxy)-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)-3-hydroxybutanamide [ka]

[0158] Step 1 (9H-Fluoren-9-yl)methyl((9S)-4-(difluoromethoxy)-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 5a 1 liter (200 mg, 0.30 mmol) of the compound was dissolved in 5 mL of DMF, and cesium carbonate (198 mg, 0.61 mmol) and (bromodifluoromethyl)phosphonate diethyl ester (121 mg, 0.45 mmol) were added under ice bath. The reaction was stirred at 0°C for 1 hour. After the reaction was complete, water was added, the system was extracted with ethyl acetate, the organic phase was washed with saturated brine, the organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain the crude product. The crude product was separated and purified by column chromatography to obtain the title compound 5a (200 mg, yield: 93%). MS m / z (ESI): 711.2 (M+1) + .

[0159] Step 2 (9S)-1-amino-4-(difluoromethoxy)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-10,13-dione 5b Compound 5a (140 mg, 0.20 mmol) was dissolved in N,N-dimethylformamide (1 mL), and diethylamine (0.1 mL) was added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the solvent in the reaction mixture was evaporated by rotation using an oil pump under vacuum to obtain the crude product 5b (90 mg) of the title compound. The crude product was used directly in the next step without purification. MS m / z (ESI): 488.1 (M+1) + .

[0160] Step 3 (R)-N-((1S,9S)-4-(difluoromethoxy)-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)-3-hydroxybutanamide (R)-N-((1R,9S)-4-(difluoromethoxy)-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)-3-hydroxybutanamide Crude product 5b (90 mg, 0.18 mmol) was dissolved in N,N-dimethylformamide (1 mL), and (R)-3-hydroxybutyric acid (23 mg, 0.22 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (105 mg, 0.28 mmol), and N,N-diisopropylethylamine (48 mg, 0.37 mmol) were added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product, which was then purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, column: Xtimate C18 250*30 mm, 10 μm; mobile phase 1: water (0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 37%~47%, flow rate: 50 mL / min) to obtain title compound 5-1 (8.6 mg, yield: 16%) and title compound 5-2 (7.8 mg, yield: 14.7%).

[0161] Single-stereoconfiguration compounds 5-1 (short retention time): MS m / z (ESI): 574.1 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.49 (d, J = 8.4 Hz, 2H), 8.06 (d, J = 11.2 Hz, 1H), 7.34 (s, 1H),7.31 (t, J = 72.8 Hz, 1H) 6.54 (s, 1H), 5.63 - 5.54 (m, 1H), 5.43 (s, 2H), 5.32 - 5.16 (m, 2H), 4.68 (d, J = 4.4 Hz, 1H), 4.05 (s, 1H), 3.23 (d, J = 5.6 Hz, 1H), 2.36 - 2.19 (m, 2H), 2.19 - 2.05 (m, 2H), 1.93 - 1.78 (m, 2H), 1.10 (d, J = 6.4 Hz, 3H), 0.88 (t, J = 7.2 Hz, 3H).

[0162] Single stereochemical compound 5-2 (long retention time): MS m / z (ESI): 574.1 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.52 (d, J = 8.8 Hz, 2H), 8.10 - 8.04 (m, 1H), 7.35 (s, 1H), 7.32 (t, J = 72.8 Hz, 1 H) 6.55 (s, 1H), 5.66 - 5.58 (m, 1H), 5.43 (d, J = 16.8 Hz, 2H), 5.26 (t, J = 12.0 Hz, 2H), 4.66 (d, J = 4.4 Hz, 1H), 4.14 - 4.01 (m, 1H), 3.25 (s, 1H), 2.34 - 2.20 (m, 2H), 2.16 - 2.07 (m, 2H), 1.92 - 1.80 (m, 2H), 1.10 (d, J = 6.4 Hz, 3H), 0.89 (t, J = 7.2 Hz, 3H).

[0163] Example 6 (R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1R,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide [ka]

[0164] Step 1: 1-Bromo-3-fluoro-2-methyl-5-nitrophenyl 6b 2-Fluoro-1-methyl-4-nitrophenyl 6a (20.0 g, 0.13 mol) was dissolved in n-heptane (50 mL), concentrated sulfuric acid (50 mL) was added, and the mixture was heated to 60°C. N-bromosuccinimide (35.6 g, 0.20 mol) was added gradually at this temperature, and the mixture was reacted at 60°C for 2 hours. The reaction mixture, cooled to room temperature, was added dropwise to ice water, extracted with toluene, and the organic phases were combined. The mixture was then washed sequentially with sodium sulfite solution, water, and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound 6b (30.0 g). The product was used directly in the next step without purification. MS m / z (ESI): 233.9 (M+1) + . 1 H NMR (400MHz, CDCl3) δ 8.29-8.23 (m, 1H), 7.88 (dd, 1H), 2.44 (d, 3H).

[0165] Step 2: 3-Bromo-5-fluoro-4-methylaniline 6c Compound 6b (30.0 g, 0.13 mol) was dissolved in methanol (200 mL), platinum-carbon (3.0 g, 5% content) was added, and the mixture was purged with hydrogen gas and reacted at room temperature for 16 hours under a hydrogen gas atmosphere. After the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated to obtain the title compound 6c (20.0 g, yellow oily substance). The product was used directly in the next step without purification. MS m / z (ESI): 204.0 (M+1) + .

[0166] Step 3: N-(3-bromo-5-fluoro-4-methylphenyl)acetamide 6d Under ice bath conditions, 3-bromo-5-fluoro-4-methylaniline 6c (20.0 g, 0.10 mol) was dissolved in dichloromethane (100 mL), triethylamine (20.2 g, 0.20 mol) and acetyl chloride (11.8 g, 0.15 mol) were added, and the mixture was reacted for 3 hours under ice bath conditions. After the reaction was complete, water was added to the reaction mixture to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was beaten with a mixed solvent of dichloromethane and petroleum ether (V / V = 10:1) to obtain the title compound 6d (10.0 g, yield: 32%). MS m / z (ESI): 246.0 (M+1) + .

[0167] Step 4 (E)-5-(5-acetamido-3-fluoro-2-methylphenyl)penta-4-enoic acid 6e N-(3-bromo-5-fluoro-4-methylphenyl)acetamide 6d (10.0 g, 40.8 mmol) was dissolved in dioxane (40 mL) and water (10 mL), and penta-4-enoic acid (6.1 g, 61.20 mmol), palladium acetate (0.7 g, 4.10 mmol), tris(o-methylphenyl)phosphine (2.5 g, 8.20 mmol), and N,N-diisopropylethylamine (15.9 g, 122.01 mmol) were added. The reaction mixture was stirred at 100 °C for 16 hours. After the reaction was complete, water and dichloromethane were added, the system was washed three times with saturated sodium bicarbonate solution, and the aqueous phase was collected. The pH of the aqueous phase was adjusted to 2-3 with hydrochloric acid, and the organic phase was collected by extracting three times with ethyl acetate, drying over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain the crude product compound 6e (10 g). The product is not purified and the reaction proceeds directly to the next step. MS m / z (ESI): 266.1 (M+1) + .

[0168] The subsequent synthesis route followed the one in Example 3, replacing intermediate 3d from Example 3 with intermediate 6e (10 g, 0.04 mol). The final product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge C18 150*19 mm, 5 μm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 37%~47%, flow rate: 20 mL / min) to obtain the title products 6-1 (12.6 mg, yield: 14%) and 6-2 (17.2 mg, yield: 19%).

[0169] Single-stereoconfiguration compounds 6-1 (short retention time): MS m / z (ESI): 536 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.60 - 8.55 (m, 1H), 7.73 - 7.72 (m, 1H), 7.28 (s, 1H), 6.51 (s, 1H), 5.54 - 5.53 (m, 1H), 5.42 (s, 2H), 5.32 - 5.26 (m, 1H), 5.24 - 5.22 (m, 1H), 4.65 (s, 1H), 4.01 (s, 1H), 3.22 - 3.18 (m, 2H), 2.42 - 2.40 (m, 3H), 2.28 - 2.24 (m, 4H), 2.10 - 1.67 (m, 6H), 1.06 (d, J = 6.0Hz, 3H), 0.85 - 0.82 (m, 3H).

[0170] Single-stereoconfiguration compounds 6-2 (long retention time): MS m / z (ESI): 536 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.67 - 8.66 (m, 1H), 7.73 - 7.71 (m, 1H), 7.28 (s, 1H), 6.49 (s, 1H), 5.53 - 5.52 (m, 1H), 5.41 - 5.40 (m, 3H), 5.35 - 5.33 (m, 1H), 4.67 - 4.65 (m, 1H), 4.03 - 3.96 (m, 1H), 3.22 - 3.18 (m, 2H), 2.42 (s, 4H), 2.40 - 2.15 (m, 4H), 2.11 - 1.52 (m, 6H), 1.08 - 1.07 (m, 3H), 0.87 - 0.85 (m, 3H).

[0171] Example 7 (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide [ka]

[0172] Step 1 (9H-Fluoren-9-yl)methyl((9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)carbamate 7a Compound 3k (140 mg, 0.20 mmol) and potassium vinyl trifluoroborate (54 mg, 0.40 mmol) were dissolved in dioxane (16 mL) and water (4 mL). Potassium phosphate (128 mg, 0.60 mmol) and methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) (34 mg, 0.04 mmol) were added, and the mixture was purged with nitrogen. The reaction mixture was stirred at 100 °C for 5 hours. After the reaction was complete, water (20 mL) was added to the reaction mixture, and it was extracted with dichloromethane (15 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 7a (80 mg, yield: 60%). MS m / z (ESI): 670.2 (M+1) + .

[0173] Step 2: Preparation of (S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-vinyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-10,13-dione Compound 7a (80 mg, 0.12 mmol) was dissolved in N,N-dimethylformamide (10 mL), and the reaction solution with diethylamine (1 mL) was added and the mixture was stirred at room temperature for 1 hour. After the reaction was complete, the mixture was concentrated under vacuum, beaten with ethyl acetate, and filtered to obtain the title compound 7b (30 mg, yield: 51%). MS m / z (ESI): 448.2 (M+1) + .

[0174] Step 3 (R)-N-((1S, 9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1R, 9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide Compound 7b (30 mg, 0.07 mmol) and (R)-3-hydroxybutyric acid (8 mg, 0.08 mmol) were dissolved in N,N-dimethylformamide (5 mL), and N,N-diisopropylethylamine (18.2 mg, 0.14 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (39.9 mg, 0.11 mmol) were added. The reaction mixture was stirred at 25°C for 5 hours. After the reaction was complete, the reaction mixture was concentrated and purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge C18 150*19mm, 5μm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 43%~53%, flow rate: 20 mL / min) to obtain title compound 7-1 (1.76 mg, yield: 10%) and title compound 7-2 (2.31 mg, yield: 10%).

[0175] Single-stereoconfiguration compounds 7-1 (short retention time): MS m / z (ESI): 534.2 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.51 (d, J = 8.8 Hz, 1H), 8.48 (s, 1H), 7.84 (d, J = 12.0 Hz, 1H), 7.32 (s, 1H), 6.97 (dd, J = 18.0, 12.0 Hz, 1H), 6.58 (s, 1H), 5.91 - 5.77 (m, 2H), 5.61 - 5.50 (m, 1H), 5.43 (s, 2H), 5.22 (q, J = 19.2 Hz, 2H), 4.72 (s, 1H), 4.04 (dd, J = 12.4, 6.4 Hz, 1H), 3.27 (d, J = 4.8 Hz, 1H), 2.30 - 2.20 (m, 2H), 2.11 (t, J = 9.2 Hz, 2H), 1.90 - 1.80 (m, 2H), 1.09 (d, J = 6.4 Hz, 3H), 0.87 (t, J = 7.2 Hz, 3H).

[0176] Single stereoconfiguration compound 7-2 (long retention time): MS m / z (ESI): 534.2 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.55 (d, J = 8.8 Hz, 1H), 8.47 (s, 1H), 7.85 (d, J = 12.0 Hz, 1H), 7.32 (s, 1H), 6.97 (dd, J = 18.0, 12.0 Hz, 1H), 6.62 (s, 1H), 5.82 (dd, J = 24.8, 14.0 Hz, 2H), 5.57 (dt, J = 8.8, 4.4 Hz, 1H), 5.43 (d, J = 1.2 Hz, 2H), 5.24 (s, 2H), 4.72 (s, 1H), 4.03 (dt, J = 12.8, 6.4 Hz, 1H), 3.27 (d, J = 5.4 Hz, 1H), 2.23 (ddd, J = 18.4, 13.2, 6.4 Hz, 2H), 2.08 (dd, J = 26.4, 10.0 Hz, 2H), 1.87 (tt, J = 14.0, 7.2 Hz, 2H), 1.08 (d, J = 6.0 Hz, 3H), 0.87 (s, 3H).

[0177] Example 8 (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-(2,2,2-trifluoroethoxy)-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-(2,2,2-trifluoroethoxy)-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide [ka]

[0178] Step 1 (9H-Fluoren-9-yl)methyl((9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-(2,2,2-trifluoroethoxy)-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)carbamate 8a 1 liter (200 mg, 0.30 mmol) of the compound was dissolved in 3 mL of N,N-dimethylformamide, and 141 mg (0.60 mmol) of 2,2,2-trifluoroethyltrifluoromethanesulfonate and 296 mg (0.90 mmol) of cesium carbonate were slowly added. The reaction mixture was stirred at room temperature for 10 minutes. After the reaction was complete, the mixture was diluted with 10 mL of water, extracted with ethyl acetate (10 mL x 3), and the organic phases were combined and concentrated. The resulting residue was purified using silica gel column chromatography with eluent system B to obtain the title compound 8a (180 mg, yield: 75%). MS m / z (ESI): 742.2 (M+H) + .

[0179] Step 2 (9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-(2,2,2-trifluoroethoxy)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-10,13-dione 8b Compound 8a (180 mg, 0.24 mmol) was dissolved in N,N-dimethylformamide (3 mL), and diethylamine (35 mg, 0.48 mmol) was added. The reaction mixture was allowed to react at room temperature for 1 hour. After the reaction was complete, the reaction mixture was concentrated under reduced pressure to obtain crude product 8b (100 mg), which was used directly in the next step without purification. MS m / z (ESI): 520.1 (M+H) + .

[0180] Step 3 (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-(2,2,2-trifluoroethoxy)-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-(2,2,2-trifluoroethoxy)-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyran[3',4',6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide Compound 8b (50 mg, 0.09 mmol) was dissolved in N,N-dimethylformamide (2 mL), and (R)-3-hydroxybutyric acid (15 mg, 0.14 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (73 mg, 0.19 mmol), and N,N-diisopropylethylamine (37 mg, 0.28 mmol) were added. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the crude product was concentrated under reduced pressure, and preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, column: Xtimate C18 150*19mm, 10μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 38%~48%, flow rate: 50 mL / min) yielded title compound 8-1 (2.5 mg, yield: 4.6%) and title compound 8-2 (3.5 mg, yield: 6.4%).

[0181] Single-stereoconfiguration compounds 8-1 (short retention time): MS m / z (ESI): 606.2 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.43 (d, J = 8.8 Hz, 1H), 7.98 (d, J = 12.0 Hz, 1H), 7.31 (s, 1H), 6.52 (s, 1H), 5.59 - 5.53 (m, 1H), 5.42 (s, 2H), 5.23 (s, 2H), 4.93 - 4.85 (m, 2H), 4.66 (d, J = 4.8 Hz, 1H), 4.08 - 4.03 (m, 1H), 3.21 (d, J = 6.0 Hz, 2H), 2.34 - 2.27 (m, 1H), 2.22 (d, J = 8.4 Hz, 1H), 2.10 (dd, J = 12.4, 5.6 Hz, 2H), 1.89 - 1.81 (m, 2H), 1.10 (d, J = 6.4 Hz, 3H), 0.90 - 0.84 (m, 3H).

[0182] Single stereochemical compound 8-2 (long retention time): MS m / z (ESI): 606.2 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.45 (d, J = 8.8 Hz, 1H), 7.98 (d, J = 12.4 Hz, 1H), 7.32 (s, 1H), 6.52 (s, 1H), 5.63 - 5.55 (m, 1H), 5.43 (s, 2H), 5.33 - 5.18 (m, 2H), 4.88 (dd, J = 18.0, 9.2 Hz, 2H), 4.65 (d, J = 4.4 Hz, 1H), 4.09 - 4.00 (m, 1H), 3.24 - 3.12 (m, 2H), 2.28 (dd, J = 13.6, 7.6 Hz, 1H), 2.19 (dd, J = 13.6, 5.6 Hz, 1H), 2.09 (dd, J = 16.8, 4.8 Hz, 2H), 1.93 - 1.78 (m, 2H), 1.09 (d, J = 6.4 Hz, 3H), 0.94 - 0.78 (m, 3H).

[0183] Example 9 (S)-9-ethyl-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzopyran[3',4',6,7]indolidino[1,2-b]quinoline-10,13-dione 9 [ka]

[0184] Step 1: 8-amino-5-methoxy-3,4-dihydronaphthalene-1(2H)-one 9a Compound 2e (500 mg, 2.14 mmol) was dissolved in a mixed solvent of methanol and concentrated hydrochloric acid (36 mL, V / V = 5:1). The reaction mixture was allowed to react at 60°C for 2 hours. After the reaction was complete, the reaction mixture was directly concentrated to obtain the title compound 9a (400 mg, yield: 88%). MS m / z (ESI): 192.1 (M+1) + .

[0185] Step 2 (S)-9-ethyl-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzopyran[3',4',6,7]indolidino[1,2-b]quinoline-10,13-dione 9b Compound 9a (400 mg, 2.09 mmol) was dissolved in toluene (15 mL), and compound 1j (661 mg, 2.51 mmol) was added. The reaction mixture was stirred at 110°C for 16 hours. After the reaction was complete, water (20 mL) was added to the reaction mixture, and it was extracted with dichloromethane (15 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. Purification using silica gel column chromatography with eluent system B yielded the title compound 9b (698 mg, yield: 80%). MS m / z (ESI): 419.1 (M+1) + .

[0186] Step 3 (S)-9-ethyl-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzopyran[3',4',6,7]indolidino[1,2-b]quinoline-10,13-dione 9 Compound 9b (100 mg, 0.24 mmol) was dissolved in hydrobromic acid (30 mL). The reaction mixture was allowed to react at 100°C for 2 hours. After the reaction was complete, the reaction mixture was directly concentrated and then purified by preparative high-performance liquid chromatography (Waters MS triggered Prep-LC with Acquity QDA detector, chromatography column: Welch C18 250 × 21.2 mm, 10 μm; mobile phase 1: water (containing 0.1% aqueous ammonia); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 10%~30%, flow rate: 25 mL / min) to obtain the title compound 9 (19.4 mg, 20%). MS m / z (ESI): 405.1 (M+1) + . 1 H NMR (400 MHz, CD3OD) δ 7.79 - 7.71 (m, 2H), 7.29 (d, J = 9.0 Hz, 1H), 5.01 (d, J = 4.0 Hz, 2H), 4.95 (d, J = 13.6 Hz, 2H), 3.01 - 2.86 (m, 4H), 2.43 - 2.35(m, 1H), 2.28 - 2.19 (m, 1H),2.06 - 2.00 (m, 2H), 1.10 - 1.00 (m, 3H).

[0187] Example 10 (1S,3R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide [ka]

[0188] Step 1 (1R, 10S)-1-amino-10-ethyl-6-fluoro-10-hydroxy-5-methyl-2,3,4,10,13,16-hexahydro-14H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-11,14-(1H)-dione (1S, 10S)-1-amino-10-ethyl-6-fluoro-10-hydroxy-5-methyl-2,3,4,10,13,16-hexahydro-14H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-11,14-(1H)-dione Compound 6m (700 mg) was fractionated by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, column: Xbridge 5 μm C18 150 x 30 mm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 22%~32%, flow rate: 50 mL / min) to obtain compound 6m-1 (290 mg, short retention time) and 6m-2 (300 mg, long retention time). MS m / z (ESI): 450.2 (M+H) +

[0189] Step 2 (1S,3R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide 10 Compound 6m-2 (30 mg, 0.06 mmol) was dissolved in N,N-dimethylformamide (3 mL), and (1S,3S)-3-hydroxycyclobutane-1-formic acid (8.5 mg, 0.07 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (38 mg, 0.10 mmol), and N,N-diisopropylethylamine (17.2 mg, 0.13 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was diluted with water (10 mL), extracted with ethyl acetate (10 mL × 3), the organic phases were combined and washed with saturated brine, dried, and concentrated to obtain the crude product. The crude product was purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, column: Xbridge 5μm C18 150 x 30mm; mobile phase 1: water (containing 0.1% TFA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 30%~95%, flow rate: 50 mL / min) to obtain compound 10 (2 mg, yield: 5%). MS m / z (ESI): 548.2 (M+H) + . 1 H NMR (400MHz, CD3OD) δ 8.71 (d, 1H), 7.57-7.48 (m, 2H), 5.65 (s, 1H), 5.55 (d, 1H), 5.35 (dd, 2H), 5.10 (d, 1H), 4.13 (dd, 1H), 2.73 (dd, 1H), 2.61-2.33 (m, 8H), 2.20-2.08 (m, 4H), 1.96 (dd, 4H), 1.02-0.97 (m, 3H).

[0190] Example 11 (R)-2-Cyclopropyl-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide (S)-2-Cyclopropyl-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide [ka]

[0191] Step 1: 2-Cyclopropyl-2-Hydroxyacetic Acid 11b 2-Cyclopropyl-2-hydroxyacetate 11a (50 mg, 0.38 mmol) was dissolved in tetrahydrofuran (6 mL), and water (0.5 mL) and lithium hydroxide (27 mg, 1.15 mmol) were added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product 2-cyclopropyl-2-hydroxyacetic acid 11b (50 mg, white solid), which was used directly in the next step without purification. MS m / z (ESI): 117.1 (M+H) + .

[0192] Step 2 (R)-2-Cyclopropyl-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide (S)-2-Cyclopropyl-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide Compound 6m-2 (50 mg, 0.11 mmol) was dissolved in N,N-dimethylformamide (2 mL), and 2-cyclopropyl-2-hydroxyacetic acid 11b (44 mg, 0.38 mmol), N,N-diisopropylethylamine (29 mg, 0.22 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (63 mg, 0.17 mmol) were added. The reaction was stirred at room temperature for 15 minutes. After the reaction was complete, preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5 μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 10 minutes, gradient ratio: acetonitrile phase 40%~50%, flow rate: 20 mL / min) was performed. The compounds were purified to obtain compound 11-1 (2.3 mg, yield: 4%) and compound 11-2 (2.5 mg, yield: 4%).

[0193] Single-stereoconfiguration compounds (compounds with short retention times) MS m / z (ESI): 548.2 (M+H) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.49 (d, 1H), 7.74 (d, 1H), 7.28 (s, 1H), 6.50 (s, 1H), 5.56 (s, 1H), 5.49-5.40 (m, 4H), 5.29 (d, 1H), 3.61 (t, 1H), 3.22 (s, 2H), 2.42 (s, 3H), 2.12-1.63 (m, 6H), 1.11 (d, 1H), 0.87 (t, 3H), 0.47-0.27 (m, 4H).

[0194] Single-stereoconfiguration compounds (compounds with long retention times) MS m / z (ESI): 548.2 (M+H) + . 1H NMR (400MHz, DMSO-d6) δ 8.43 (d, 1H), 7.74 (d, 1H), 7.28 (s, 1H), 6.51 (s, 1H), 5.54 (s, 1H), 5.42 (t, 4H), 5.31 (d, 1H), 3.68-3.53 (m, 1H), 3.22 (s, 2H), 2.42 (s, 3H), 2.12-1.66 (m, 6H), 1.10 (d, 1H), 0.87 (t, 3H), 0.48-0.28 (m, 4H).

[0195] Example 12 (1S, 3R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide [ka]

[0196] Step 1 (1R,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-vinyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-vinyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione Compound 7b (500 mg) was fractionated by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, column: Xbridge 5 μm C18 150 x 30 mm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 22%~32%, flow rate: 50 mL / min) to obtain compounds 7b-1 (180 mg, short retention time) and 7b-2 (195 mg, long retention time). MS m / z (ESI): 448.2 (M+H) + .

[0197] Step 2 (1S, 3R)-N-((1S, 9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide 12 Compound 7b-2 (22 mg, 0.05 mmol) was dissolved in N,N-dimethylformamide (0.5 mL), and (1S, 3S)-3-hydroxycyclobutane-1-formic acid (7 mg, 0.06 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (30 mg, 0.08 mmol), and N,N-diisopropylethylamine (13 mg, 0.10 mmol) were added. The reaction was stirred at room temperature for 15 minutes. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5μm C18 150 x 19mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 10 minutes, gradient ratio: acetonitrile phase 45%~100%, flow rate: 20 mL / min) to obtain compound 12 (3 mg, yield: 11%). MS m / z (ESI): 546.2 (M+H) + . 1 H NMR (400MHz, CD3OD) δ 8.50 (s, 1H), 7.71 (d, 1H), 7.64 (s, 1H), 6.95 (dd, 1H), 5.83 (dd, 2H), 5.67-5.61 (m, 1H), 5.57 (d, 1H), 5.39 (d, 1H), 5.24 (d, 2H), 4.16-4.05 (m,1H), 3.45-3.35 (m, 2H), 2.73-2.42 (m, 4H), 2.29-2.17 (m, 4H), 2.01-1.91 (m, 2H), 1.00 (t, 3H).

[0198] Example 13 N-((7S,15R)-7-benzyl-17-(((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxo-3,6,9,12-tetraazaheptadecyl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide 13 [ka]

[0199] Step 1 (R)-3-hydroxybutyrate benzyl ester 13b (R)-3-hydroxybutyric acid 13a (50 g, 0.48 mol) was dissolved in N,N-dimethylformamide (100 mL), benzyl bromide (90 g, 0.52 mol) and cesium carbonate (312.8 g, 0.96 mol) were added, and the reaction mixture was allowed to react at room temperature for 12 hours. After the reaction was complete, water (1000 mL) was added to the reaction mixture, and it was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. The crude product was purified by silica gel column chromatography system B to obtain the title compound 13b (40 g, yield: 40%). MS m / z (ESI): 217.1 (M+Na) + .

[0200] Step 2 (R)-1-(9H-fluoren-9-yl)-10-methyl-3,6-dioxo-2,9-dioxa-4,7-diazadodecane-12-ate benzyl ester 13d (2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)acetamido)methyl acetate 13c (15 g, 40.6 mmol) was dissolved in dichloromethane (150 mL), and compound 13b (39 g, 203.2 mmol) and pyridine 4-methylbenzenesulfonate (2 g, 8.1 mmol) were added. The reaction mixture was allowed to react at 40°C for 12 hours. After the reaction was complete, the mixture was concentrated under reduced pressure, and the resulting concentrate was purified by silica gel column chromatography system B to obtain the title compound 13d (10 g, yield: 50%). MS m / z (ESI): 525.2 (M+Na) + .

[0201] Step 3 (R)-3-((2-aminoacetamide)methoxy)benzyl butyrate 13e Compound 13d (10 g, 19.9 mmol) was dissolved in N,N-dimethylformamide (50 mL), diethylamine (3 g, 39.8 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the mixture was concentrated to obtain crude product 13e (5 g), which was used directly in the next step without purification. MS m / z (ESI): 281.1 (M+H) + .

[0202] Step 4 Benzyl(5S,13R)-5-benzyl-1-(9H-fluoren-9-yl)-13-methyl-3,6,9-trioxy-2,12-dioxy-4,7,10-triazopentadecane-15-carboxylic acid 13f Compound 13e (5 g, 17.8 mmol) was dissolved in N,N-dimethylformamide (20 mL), and (S)-2,5-dioxopyrrolidone-1-yl2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-phenylpropionate (7 g, 19.5 mmol) and N,N-diisopropylethylamine (4.5 g, 35.6 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the mixture was concentrated to obtain crude product 13f (10 g), which was used directly in the next step without purification. MS m / z (ESI): 672.3 (M+Na) + .

[0203] Step 5 (R)-3-((2-((S)-2-amino-3-phenylpropionamide)acetamide)methoxy)benzyl butyrate 13g Compound 13f (10 g, 15.4 mmol) was dissolved in N,N-dimethylformamide (50 mL), and diethylamine (2 g, 30.8 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the system was diluted with water (100 mL), extracted with ethyl acetate (100 mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting concentrate was purified by silica gel column chromatography system A to obtain 13 g of product (4 g, yield: 61%). MS m / z (ESI): 428.1 (M+H) + .

[0204] Step 6 (11S,19R)-11-benzyl-1-(9H-fluoren-9-yl)-19-methyl-3,6,9,12,15-pentaoxo-2,18-dioxo-4,7,10,13,16-pentazabenzofuran-21-ate benzyl ester 13h 13 g (4 g, 9.3 mmol) of the compound was dissolved in N,N-dimethylformamide (30 mL), and (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycine (3.6 g, 10.2 mmol), N,N,N',N'-tetramethyl-O-(7-azabenzotriazole-1-yl)hexafluorophosphate urea (5.3 g, 13.9 mmol), and N,N-diisopropylethylamine (2.3 g, 18.6 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was diluted with water (100 mL), extracted with ethyl acetate (50 mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting crude product was purified by silica gel column chromatography system A to obtain the title compound 13h (3 g, yield: 42%). MS m / z (ESI): 786.3 (M+H) + .

[0205] Step 7 (11S,19R)-11-benzyl-1-(9H-fluoren-9-yl)-19-methyl-3,6,9,12,15-pentaoxo-2,18-dioxo-4,7,10,13,16-pentazabenzofuran-21-carboxylic acid 13i Compound 13h (3 g, 3.9 mmol) was dissolved in ethanol (20 mL) and ethyl acetate (10 mL), palladium / carbon (2 g, 19.5 mmol) was added, the mixture was purged with hydrogen gas, and the reaction mixture was stirred for 2 hours under a hydrogen gas atmosphere at room temperature. After the reaction was complete, the mixture was filtered through diatomaceous earth, the filter cake was washed with ethyl acetate, the filtrates were combined, and the mixture was concentrated under reduced pressure to obtain the crude product of the title compound 13i (2 g). The product was used directly in the next step without purification. MS m / z (ESI): 696.3 (M+H) + .

[0206] Step 8 (7S,15R)-1-amino-7-benzyl-15-methyl-2,5,8,11-tetraoxo-14-oxo-3,6,9,12-tetraazaheptadecane-17-carboxylic acid 13j Compound 13i (2 g, 3 mmol) was dissolved in N,N-dimethylformamide (20 mL), diethylamine (0.44 g, 6 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, tetrahydrofuran (20 mL) was added to the reaction mixture, the precipitated solid was filtered, and the filter cake was collected to obtain the title compound 13j (1 g, yield: 76%). MS m / z (ESI): 452.2 (M+H) + .

[0207] Step 9 (3R,11S)-11-benzyl-3-methyl-24-(2-(methylsulfonyl)pyrimidine-5-yl)-7,10,13,16,19-pentaoxo-4-oxo-6,9,12,15,18-pentazatetracarbonyl-23-enoic acid 13l Compound 13j (300 mg, 0.66 mmol) was dissolved in N,N-dimethylformamide (10 mL), and 2,5-dioxopyrrolidone-1-yl-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inoate (267 mg, 0.72 mmol) and N,N-diisopropylethylamine (171 mg, 1.32 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was diluted with water (50 mL), extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting crude product was purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, chromatography column: Xtimate 10 μm C18 250 x 30 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 25%~95%, flow rate: 50 mL / min) to obtain 13 L (100 mg, 21%) of the title compound. MS m / z (ESI): 724.3 (M+Na) + .

[0208] Step 10 N-((7S,15R)-7-benzyl-17-(((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxo-3,6,9,12-tetraazaheptadecyl)-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide 13 13L (100 mg, 0.14 mmol) of compound was dissolved in N,N-dimethylformamide (6 mL), and compound 6m-2 (64 mg, 0.14 mmol), 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholine hydrochloride (84 mg, 0.28 mmol), and triethylamine (29 mg, 0.28 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was diluted with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5 μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 40%~100%, flow rate: 20 mL / min) to obtain the title compound 13 (10 mg, yield: 6%). MS m / z (ESI): 1133.3 (M+H) + . 1H NMR (400 MHz, CD3OD) δ 8.92 (s, 1H), 7.60 (d, 2H), 7.16 (dd, 6H), 5.57 (d, 5H), 5.36 (s, 2H), 4.70 (d, 2H), 4.57 (d, 2H), 4.45 (d, 2H), 3.80-3.72 (m, 4H), 3.48 (s, 2H), 3.13 (s, 2H), 2.60-2.51 (m, 4H), 2.47-2.40 (m, 6H), 1.93 (d, 6H), 1.30 (d, 2H), 1.23 (d, 3H), 1.01-0.96 (m, 3H).

[0209] Example 14 N-((7S,15R)-7-benzyl-17-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexaneamide [ka]

[0210] Step 1 (3R, 11S)-11-benzyl-24-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-3-methyl-7,10,13,16,19-pentaoxo-4-oxa-6,9,12,15,18-pentazatetradecanoic acid 14a Compound 13j (200 mg, 0.44 mmol) was dissolved in N,N-dimethylformamide (2 mL), and 2,5-dioxopyrrolidone-1-yl 6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexanoate (136 mg, 0.44 mmol) and N,N-diisopropylethylamine (114 mg, 0.88 mmol) were added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: Xbridge 5 μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 10 minutes, gradient ratio: acetonitrile phase 25%~35%, flow rate: 25 mL / min) to obtain the title compound 14a (110 mg, yield: 39%). MS m / z (ESI): 667.1 (M+Na) + .

[0211] Step 2 N-((7S,15R)-7-benzyl-17-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexaneamide 14 Compound 6m-2 (50 mg, 0.11 mmol) was dissolved in N,N-dimethylformamide (0.5 mL), and compound 13b (72 mg, 0.11 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (62 mg, 0.08 mmol), and N,N-diisopropylethylamine (28 mg, 0.22 mmol) were added. The reaction was stirred at room temperature for 15 minutes. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 12 minutes, gradient ratio: acetonitrile phase 41%~51%, flow rate: 20 mL / min) to obtain compound 14 (46 mg, yield: 39%). MS m / z (ESI): 1076.3 (M+H) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.65 (d, 1H), 8.47 (s, 1H), 8.26 (d, 1H), 8.09 (d, 1H), 8.05 (s, 1H), 7.98 (d, 1H), 7.30 (s, 1H), 7.22-7.16 (m, 5H), 6.99 (s, 2H), 5.53 (s, 1H), 5.42 (s, 2H), 5.29 (s, 2H), 4.57 (d, 1H), 4.54-4.44 (m, 2H), 3.99 (d, 1H), 3.73-3.55 (m, 8H), 3.02 (d, 1H), 2.80-2.72 (m, 1H), 2.69-2.54 (m, 4H), 2.45-2.31(m, 4H), 2.09 (t, 3H), 1.94-1.76 (m, 4H), 1.46 (dd, 4H), 1.23-1.15 (m, 2H), 1.13 (d, 3H), 0.86 (t, 3H).

[0212] Example 15 N-((7S,15R)-7-benzyl-17-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexaneamide [ka]

[0213] Step 1 N-((7S,15R)-7-benzyl-17-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexanamide 15 (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-vinyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-10,13dione 7b-2 (20 mg, 0.04 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol), and N,N-diisopropylethylamine (11 mg, 0.09 mmol). The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5 μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 12 minutes, gradient ratio: acetonitrile phase 44%~54%, flow rate: 20 mL / min) to obtain compound 15 (6 mg, yield: 14%). MS m / z (ESI): 1074.3 (M+1) + . 1 H NMR (400 MHz, CD3OD) δ 7.57-7.50 (m, 2H), 7.19-7.09 (m, 6H), 6.93 (dd, 1H), 6.76 (s, 2H), 5.81 (dd, 2H), 5.66 (t, 1H), 5.55 (d, 1H), 5.37-5.26 (m, 3H), 4.70 (d, 2H), 4.50 (dd, 1H), 4.27 (d, 1H), 3.77 (q, 4H), 3.45 (dd, 2H), 3.10 (dd, 2H), 2.88-2.81 (m, 1H), 2.51-2.45 (m, 2H), 2.30-2.18 (m, 5H), 1.89 (q, 2H), 1.57 (t, 5H), 1.28 (d, 6H), 0.98 (t, 3H).

[0214] Example 16 (1R,3R)-3-(((S)-7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-3,6,9,12,15-pentaoxo-2,7,8,11,14-pentaazanicarbonyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indridine[1,2-b]quinoline-1-yl)cyclobutan-1-carboxamide [ka]

[0215] Using the same synthetic route as in Example 13, the starting material (R)-3-hydroxybutyric acid in Step 1 was replaced with (1S,3S)-3-hydroxycyclobutane-1-carboxylic acid 16a (10 g, 86.20 mmol), and after an 8-step reaction, compound 16i (2.8 g, yield: 30%) was obtained. MS m / z (ESI): 464.2 (M+1) + .

[0216] Step 9 (1R,3S)-3-(((S)-7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-3,6,9,12,15-pentaoxo-2,7,8,11,14-pentazaeicosyl)oxy)cyclobutan-1-carboxylic acid 16j Compound 16i (750 mg, 1.62 mmol) was dissolved in DMF (5 mL), and 2,5-dioxopyrrolidone-1-yl 6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexanoate (499 mg, 1.62 mmol) and N,N-diisopropylethylamine (209 mg, 1.62 mmol) were added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, chromatography column: Xtimate 10 μm C18 250 x 30 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 20%~95%, flow rate: 50 mL / min) to obtain compound 16j (150 mg, yield: 13%). MS m / z (ESI): 679.2 (M+23) + .

[0217] Step 10 (1R,3R)-3-(((S)-7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-3,6,9,12,15-pentaoxo-2,7,8,11,14-pentaazanicarbonyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidine[1,2-b]quinoline-1-yl)cyclobutan-1-carboxamide 16 Compound 7b-2 (20 mg, 0.04 mmol), compound 16j (33 mg, 0.05 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol), and N,N-diisopropylethylamine (11 mg, 0.09 mmol). The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5 μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 12 minutes, gradient ratio: acetonitrile phase 40%~55%, flow rate: 20 mL / min) to obtain compound 16 (10 mg, yield: 23%). MS m / z (ESI): 1086.2 (M+H) + . 1 H NMR (400 MHz, CDOD) δ 8.52 (s, 2H), 7.71 (d,1H), 7.63 (s, 1H), 7.29-7.15 (m, 4H), 6.94 (dd, 1H), 6.76 (s, 1H), 5.90-5.78 (m, 2H), 5.64 (s, 1H), 5.54 (d, 1H), 5.35 (d, 1H), 5.24 (d, 1H), 4.72-4.63 (m, 2H), 4.49-4.44 (m, 1H), 4.04 (s, 1H), 3.96-3.85 (m, 2H), 3.83-3.69 (m, 4H), 3.49-3.46 (m, 1H), 3.46-3.41 (m, 2H), 3.15 (s, 1H), 3.14-3.11 (m, 1H), 2.96 (s, 1H), 2.70 (s, 1H), 2.66-2.54 (m, 2H), 2.50-2.45 (m, 1H), 2.34-2.26 (m, 2H), 2.26-2.17 (m, 3H), 1.99-1.91 (m, 2H), 1.63-1.48 (m, 4H), 1.28 (d, 3H), 1.00 (t, 3H).

[0218] Example 17 (R)-N-((1R,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide [ka]

[0219] Step 1: N-(4-cyclopropyl-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 17a Compound 3f (1.5 g, 5.02 mmol), cyclopropylboronic acid (1.3 g, 15.06 mmol), 1,1-bis(diphenylphosphin)ferrocenepalladium chloride (2.2 g, 3.01 mmol), and cesium carbonate (1.6 g, 15.06 mmol) were dissolved in dioxane (15 mL). The reaction was carried out in a microwave reactor at 110 °C for 2 hours with stirring. After the reaction was complete, the reaction mixture was poured into water, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and the organic phases were combined and concentrated. Purification by silica gel column chromatography system B was obtained to obtain the title compound 17a (1.1 g, yield: 84%). MS m / z (ESI): 262.1 (M+1) + .

[0220] Step 2 (Z) -N-(4-cyclopropyl-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 17b Potassium tert-butoxide (0.94 g, 8.4 mmol) was dissolved in a mixed solvent of tetrahydrofuran and tert-butanol (50 mL, V / V = 4:1), and compound 17a (1.1 g, 4.2 mmol) was dissolved in 10 mL of tetrahydrofuran solution. This mixture was slowly added to the reaction system at 0°C. The reaction was stirred at 0°C for 10 minutes, after which isoamyl nitrite (0.74 g, 6.3 mmol) was added, and the reaction was continued with stirring at 0°C for 50 minutes. After the reaction was complete, dilute hydrochloric acid was added to adjust the pH of the system to 4-5, and the organic phase was extracted three times with ethyl acetate. The organic phase was washed with saturated brine, collected, dried over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain crude product 17b. The product was used directly in the next step without purification. MS m / z (ESI): 291.0 (M+1) + .

[0221] Step 3 N-(7-amino-4-cyclopropyl-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide 17c Compound 17b was added to methanol (50 mL), and palladium-carbon catalyst (500 mg) was added. The reaction was stirred at room temperature for 2 hours under a hydrogen gas environment. After the reaction was complete, the mixture was filtered, and the filtrate was collected to obtain crude product 17c. The crude product was used directly in the next step without purification. MS m / z (ESI): 277.1 (M+1) + .

[0222] Step 4 (9H-Fluoren-9-yl)methyl(8-acetamido-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate 17d After adjusting the pH of the filtrate obtained in the previous step to 8-9 with saturated sodium carbonate solution, Fmoc-Cl (1.19 g, 4.6 mmol) was added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the system was extracted with ethyl acetate, the organic phase was washed with saturated brine, the organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain the crude product. Compound 17d (1 g, yield: 48%) was obtained from the crude product using silica gel column chromatography system A. MS m / z (ESI): 499.1 (M+1) + .

[0223] Step 5 (9H-fluoren-9-yl)methyl(8-amino-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)carbamate 17e Compound 17e (1 g, 2.01 mmol) was dissolved in dioxane (20 mL) to obtain concentrated hydrochloric acid (12 mol / L, 5 mL). The reaction was stirred at 60°C for 1 hour. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product, which was purified by silica gel column chromatography system A to obtain compound 17f (700 mg, yield: 77%). MS m / z (ESI): 457.1 (M+1) + .

[0224] Step 6 (9H-Fluoren-9-yl)methyl((9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)carbamate 17g Compound 17f (700 mg, 1.54 mmol) was dissolved in toluene (10 mL), and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f]indolidine-3,6,10(4H)-trione (484 mg, 1.84 mmol) and p-toluenesulfonic acid monohydrate (292 mg, 1.54 mmol) were added. The reaction system was stirred at 140 °C for 4 hours. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product, which was purified by silica gel column chromatography system A to obtain 17 g (700 mg, yield: 66%) of the title compound. MS m / z (ESI): 684.2 (M+1) + .

[0225] Step 7 (9S)-1-amino-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[d]pyran[3',4':6,7indolidino[1,2-b]quinoline-10,13-dione 17h 17 g (100 mg, 0.15 mmol) of the compound was dissolved in N,N-dimethylformamide (5 mL), and diethylamine (0.5 mL) was added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the mixture was concentrated under vacuum to obtain 17 h (60 mg) of the crude product compound. The crude product was not purified, and the next step in the reaction was carried out directly. MS m / z (ESI): 462.1 (M+1) + .

[0226] Step 8 (R)-N-((1R,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide Compound 17h (60 mg, 0.13 mmol) was dissolved in DMF (5 mL), and then (R)-3-hydroxybutyric acid (19 mg, 0.18 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (87 mg, 0.23 mmol), and N,N-dimethylacetamide (59 mg, 0.45 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5μm C18 150 x 19mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 38%~48%, flow rate: 20 mL / min) to obtain compounds 17-1 (6 mg, yield: 11%) and 17-2 (4 mg, yield: 8%).

[0227] Single-stereoconfiguration compound 17-1 (short retention time): MS m / z (ESI): 548.2 (M+1) + . 1H NMR (400MHz, DMSO-d6) δ 8.44 (d, 1H), 7.75 (d, 1H), 7.47 (d, 1H), 7.30 (s, 1H), 7.11 (d, 1H), 5.60-5.52 (m, 1H), 5.43 (s, 2H), 5.29-5.13 (m, 2H), 4.09-4.00 (m, 1H), 2.48 (s, 1H), 2.29 (s, 2H), 2.24-2.18 (m, 1H), 2.14 (d, 2H), 1.99-1.92 (m, 1H), 1.91-1.79 (m, 2H), 1.15-1.10 (m, 2H), 1.09 (d, 3H), 0.87 (s, 3H), 0.76 (s, 2H).

[0228] Single-stereoconfiguration compounds 17-2 (long retention time): MS m / z (ESI): 548.2 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (d, 1H), 7.76 (d, 1H), 7.47 (d, 1H), 7.30 (s, 1H), 7.13-7.06 (m, 1H), 5.59 (dd, 1H), 5.43 (d, 2H), 5.24 (d, 2H), 4.05 (dd, 1H), 2.29 (t, 1H), 2.25 (d, 1H), 2.19 (d, 1H), 2.18-2.06 (m, 3H), 1.95 (d, 1H), 1.85 (dd, 2H), 1.16-1.10 (m, 2H), 1.08 (d, 3H), 0.87 (s, 4H), 0.76 (d, 2H).

[0229] Example 18 (1S,3S)-N-((1R,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide (1S,3R)-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide [ka]

[0230] Step 1 (1S,3S)-N-((1R,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide (1S,3R)-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide

[0231] 17 g (69 mg, 0.15 mmol) of the compound was dissolved in N,N-dimethylformamide (5 mL), and (1S, 3S)-3-hydroxycyclobutane-1-formic acid (21 mg, 0.18 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (87 mg, 0.23 mmol), and N,N-dimethylacetamide (59 mg, 0.45 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 18 minutes, gradient ratio: acetonitrile phase 35%~65%, flow rate: 20 mL / min) to obtain 18-1 (4 mg, yield: 6%) and 18-2 (2.4 mg, yield: 4%).

[0232] Single-stereoconfiguration compounds 18-1 (short retention time): MS m / z (ESI): 560.2 (M+1) + . 1 H NMR (400MHz, DMSO-d6) δ 8.44 (d, 1H), 7.71 (d, 1H), 7.29 (d, 1H), 6.54 (d, 1H), 5.56 (s, 1H), 5.42 (s, 2H), 5.17-5.09 (m, 3H), 3.94 (d, 1H), 2.44-2.30 (m, 4H), 2.08-2.03 (m, 4H), 1.94-1.84 (m, 4H), 1.11 (d, 2H), 0.87 (t, 3H), 0.76 (s, 2H).

[0233] Single-stereoconfiguration compounds 18-2 (long retention time): MS m / z (ESI): 560.2 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.45 (d, 1H), 7.74 (d, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.57 (s, 1H), 5.43 (s, 2H), 5.14 (dd, 3H), 3.97-3.91 (m, 1H), 2.40-2.27 (m, 4H), 2.15-2.05 (m, 4H), 1.96-1.84 (m, 4H), 1.11 (d, 2H), 0.87 (t, 3H), 0.75 (d, 2H).

[0234] Example 19 (R)-N-((1S, 10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1R, 10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxybutanamide [ka]

[0235] Step 1: 5-(5-acetamido-2-bromo-3-fluorophenyl)penta-4-enoic acid 19b Compound 19a (7.0 g, 19.5 mmol) was dissolved in dioxane (60 mL) and water (15 mL). Penta-4-enoic acid (2.2 g, 22.0 mol), bistriphenylphosphine palladium dichloride (700 mg, 1.0 mmol), and sodium carbonate (6.2 g, 58.4 mmol) were added sequentially to the solution. The reaction mixture was stirred at 90°C for 16 hours under nitrogen gas protection. After the reaction was complete, the reaction mixture was filtered through diatomaceous earth and washed with ethyl acetate. The resulting filtrate was directly concentrated. The crude product was diluted with water (200 mL), extracted three times with ether (100 mL x 3), the aqueous phase was adjusted to pH=2 with dilute hydrochloric acid, extracted with ethyl acetate (100 mL x 3), the organic phases were combined and concentrated to obtain crude product 19b (6.0 g). MS m / z (ESI): 330.1 (M+1) + .

[0236] Step 2 5-(5-acetamido-2-bromo-3-fluorophenyl)pentaic acid 19c Compound 19b (6.0 g, 18.2 mmol) was dissolved in anhydrous methanol (100 mL), and platinum / carbon (5%, 600 mg) was added. The reaction mixture was purged three times with hydrogen gas and stirred at room temperature for 16 hours. After the reaction was complete, the reaction mixture was filtered through diatomaceous earth and washed with ethyl acetate. The resulting filtrate was directly concentrated to obtain the crude product compound 19c (5.0 g). MS m / z (ESI): 332.0 (M+1) + .

[0237] Step 3 N-(4-bromo-3-fluoro-9-oxo-6,7,8,9-tetrahydro-5H-benzo[7]cycloalken-1-yl)acetamide 19d Compound 19c (2.5 g, 3.3 mmol) was dissolved in polyphosphate (25 mL). The reaction mixture was stirred at 140°C for 2 hours. After the reaction was complete, water (100 mL) was added to the reaction mixture and stirred for 4 hours. The aqueous phase was extracted with ethyl acetate (100 mL x 3), the organic phase was combined, and the resulting crude product was concentrated and purified by silica gel column chromatography system B to obtain compound 19d (920 mg, yield: 39%). MS m / z (ESI): 314.0 (M+1) + .

[0238] The subsequent synthesis route was the same as in Example 17, but compound 3f was replaced with 19d (720 mg, 2.3 mmol). The final product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 40%~95%, flow rate: 30 mL / min) to obtain products 19-1 (6.5 mg, yield: 9%) and 19-2 (11 mg, yield: 16%).

[0239] Single-stereoconfiguration compounds 19-1 (short retention time): MS m / z (ESI): 562.2 (M+1) + . 1H NMR (400MHz, CDCl3) δ 7.63-7.50 (m, 3H), 7.07 (s, 1H), 5.73-5.69 (m, 2H), 5.43 (d, 1H), 5.36-5.15 (m, 2H), 4.24 (s, 1H), 3.81 (s, 1H),3.64-3.50 (m, 1H), 3.47-3.32 (m, 1H), 2.52-2.41 (m, 2H), 2.36 -2.12 (m, 3H), 1.88-1.83 (m, 4H), 1.25 (d, 3H), 1.19-1.15 (m, 2H), 1.02 (t, 3H), 0.81-0.77 (m, 2H).

[0240] Single-stereoconfiguration compounds 19-2 (long retention time): MS m / z (ESI): 562.2 (M+1) + . 1 H NMR (400 MHz, CDCl3) δ 7.50-7.35 (m, 3H), 5.72-5.55 (m, 1H), 5.47-5.29 (m, 2H), 5.26-5.05 (m, 2H), 4.51-4.31 (m, 1H), 4.05-3.91 (m, 1H), 3.70-3.54 (m, 1H), 3.53-3.37 (m, 1H), 2.66-2.50 (m, 2H), 2.49-2.37 (m, 1H), 2.28-2.14 (m, 1H), 2.11-1.75 (m, 5H), 1.41-1.29 (m, 3H), 1.29-1.10 (m, 2H), 1.04-0.95 (m, 3H), 0.88-0.71 (m, 2H).

[0241] Example 20 (1S,3S)-N-((1R,10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide (1S,3S)-N-((1S,10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide [ka]

[0242] Step 1 (1S,3S)-N-((1R,10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide (1S,3S)-N-((1S,10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide

[0243] Compound 19k (75 mg, 0.16 mmol) was dissolved in N,N-dimethylformamide (3 mL), and (1S,3S)-3-hydroxycyclobutane-1-formic acid (22 mg, 0.19 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (91 mg, 0.24 mmol), and N,N-diisopropylethylamine (171 mg, 1.32 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was diluted with water (50 mL), extracted with ethyl acetate (30 mL x 3), washed with saturated brine after combining the organic phases, dried, concentrated, and the crude product was purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% NH3.H2O); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 40%~100%, flow rate: 30 mL / min) to obtain compounds 20-1 (2.9 mg, 7%) and 20-2 (1.5 mg, 2%).

[0244] Single-stereoconfiguration compounds 20-1 (short retention time): MS m / z (ESI): 574.7 (M+1) + . 1H NMR (400MHz, CDCl3) δ 7.59 (d, 1H), 7.53 (s, 1H), 6.49 (s, 1H), 5.72-5.64 (m, 1H), 5.59 (d, 1H), 5.35 (d, 1H), 5.14 (d, 1H), 4.93 (d, 1H), 4.25-4.19(m, 1H), 3.77 (s, 1H), 3.64-3.52 (m, 1H), 3.46 -3.41 (m, 1H), 2.77-2.68 (m, , 3H), 2.42 -2.10 (m, 5H), 1.94-1.78 (m, 4H), 1.25-1.75 (m, 2H), 1.05 (t, 3H), 0.87-0.72 (m, 2H).

[0245] Single-stereoconfiguration compounds 20-2 (long retention time): MS m / z (ESI): 574.7 (M+1) + . 1 H NMR (400 MHz, CDCl3) δ 7.53 (d, 1H), 7.46 (s, 1H), 6.91 (br, 1H), 5.59 (t, 1H), 5.30-5.19(m, 2H), 5.11-5.04 (m, 2H), 4.30-4.21 (m, 1H), 3.76 (s, 1H), 3.68 (d, 1H), 3.48-3.34 (m, 1H), 2.88-2.75 (m, 3H), 2.37-2.21 (m, 5H), 1.93 (s, 3H), 1.80-1.72 (m, 2H), 1.23-1.13 (m, 2H), 0.96 (t, 3H), 0.87-0.75 (m, 2H).

[0246] Example 21 N-((7S,15R)-7-benzyl-17-(((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxy-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexaneamide [ka]

[0247] Step 1 (9H-Fluoren-9-yl)methyl((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)carbamate 21a Compound 17f (600 mg, 1.02 mmol) was separated for chiral isomers using a Gilson preparative and a Daicel chiral column, and then purified using a column (CHIRALPAK IB 3.0 cm ID × 25 cm, 10 μm; mobile phase 1: MeOH; mobile phase 2: DCM; gradient mixing ratio: MeOH / DCM = 90 / 10, flow rate: 25 mL / min) to obtain compound 21a (250 mg, yield: 42%).

[0248] Step 2 (1S,9S)-1-amino-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[d]pyran[3',4':6,7]indolidino[1,2-b]quinoline-10,13-dione 21b Compound 21a (50 mg, 0.07 mmol) was dissolved in DMF (5 mL), and diethylamine (0.5 mL) was added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the solvent was evaporated under vacuum by rotation to obtain the title compound 21b (34 mg), which was used directly in the next step without purification. MS m / z (ESI): 462.1 (M+1) + .

[0249] Step 3 N-((7S,15R)-7-benzyl-17-(((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxy-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexanamide 21 Compound 21b (34 mg) was dissolved in DMF (2 mL), and compound 14a (54 mg, 0.08 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (40 mg, 0.11 mmol), and N,N-diisopropylethylamine (18 mg, 0.14 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: Xbridge 5 μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 12 minutes, gradient ratio: acetonitrile phase 41%~51%, flow rate: 25 mL / min) to obtain compound 21 (20 mg, yield: 26%). MS m / z (ESI): 1088.3 (M+H) + . 1 H NMR (400 MHz, CD3OD) δ 7.70 (d, 1H), 7.62 (t, 2H), 7.23-7.13 (m, 8H), 6.76 (s, 2H), 5.60 (d, 2H), 5.55 (s, 1H), 5.38 (dd, 3H), 5.23 (d, 2H), 4.41 (dd, 2H), 4.10 (d, 2H), 3.80 (dd, 5H), 3.43 (d, 2H), 3.05 (d, 1H), 3.04-2.96 (m, 2H), 2.56-2.42 (m, 4H), 2.28 (d, 2H), 2.21 (d, 2H), 1.93 (dd, 2H), 1.60 1.52 (m, 4H), 1.26 (d, 4H), 1.14 (d, 3H), 0.99 (t, 3H).

[0250] Example 22 (1R,3R)-3-(((S)-7-benzyl-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-3,6,9,12,15-pentaoxo-2,7,8,11,14-pentaazanicarbonyl)oxy)-N-((1S,9S)-4-cyclopropyl-9-fluoro-9-hydroxy-10,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)cyclobutanecarboxamide [ka]

[0251] Step 1 (1R,3R)-3-(((S)-7-benzyl-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-3,6,9,12,15-pentaoxo-2,7,8,11,14-pentaazanicarbonyl)oxy)-N-((1S,9S)-4-cyclopropyl-9-fluoro-9-hydroxy-10,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)cyclobutanecarboxamide Compound 21b (35 mg, 0.08 mmol) was dissolved in N,N-dimethylformamide (3 mL), and compound 16j (60 mg, 0.09 mmol), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (43 mg, 0.11 mmol), and N,N-diisopropylethylamine (24 mg, 0.19 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge C18 5μm 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 42%~100%, flow rate: 20 mL / min) to obtain compound 22 (20 mg, yield: 22%). MS m / z (ESI): 1100.5 (M+1) + . 1H NMR (400 MHz, CD3OD) δ 8.51 (d, 1H), 8.37-8.20 (m, 2H), 7.59-7.52 (m, 1H), 7.29-7.11 (m, 4H), 6.75 (s, 1H), 5.66 (d, 1H), 5.58-5.48 (m, 1H), 5.37-5.14 (m, 2H), 5.09-4.94 (m, 1H), 4.73-4.61 (m, 2H), 4.48 (dd, 1H), 4.09-3.97 (m, 1H), 3.97-3.68 (m, 5H), 3.57-3.46 (m, 1H), 3.45-3.37 (m, 2H), 3.15 (dd, 1H), 3.04-2.91 (m, 1H), 2.75-2.53 (m, 2H), 2.52-2.41 (m, 1H), 2.38-2.15 (m, 5H), 2.01 (dd, 1H), 1.94 -1.85 (m, 2H), 1.68-1.44 (m, 4H), 1.38-1.20 (m, 6H), 1.15 (d, 2H), 1.02-0.86 (m, 3H), 0.81 (d, 2H).

[0252] Example 23 N-((7S,15R)-7-benzyl-17-(((1S,10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxy-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexaneamide [ka]

[0253] Step 1 (9H-fluoro-9-yl)methyl((1S,10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)carbamate 23a Compound 19j (500 mg, 0.72 mmol) was separated for chiral isomers using a Gilson preparative and a Daicel chiral column, and then purified to obtain compound 23a (220 mg, yield: 44%) using a column (CHIRALPAK IA 3.0 cm ID × 25 cm, 10 μm; mobile phase 1: MeOH; mobile phase 2: DCM; gradient mixing ratio: MeOH / DCM = 70 / 30, flow rate: 25 mL / min).

[0254] Step 2 (1S,10S)-1-amino-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-2,3,4,10,13,16-hexahydro-14H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-11,14(1H)-dione 23b Compound 23a (45 mg, 0.064 mmol) was dissolved in a mixed solvent of N,N-dimethylformamide and ethylenediamine (7 mL, V / V = 5:2), and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was directly concentrated to obtain the crude compound product 23b (30 mg), which was used directly in the next step without purification. MS m / z (ESI): 476.2 (M+1) + .

[0255] Step 3 N-((7S,15R)-7-benzyl-17-(((1S,10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxy-3,6,9,12-tetraazaheptadecyl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexaneamide 23 Compound 23b (30 mg) was dissolved in N,N-dimethylformamide (5 mL), and compound 14a (43 mg, 0.064 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (37 mg, 0.096 mmol), and N,N-diisopropylethylamine (171 mg, 1.32 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, water (50 mL) was added to the reaction mixture, and it was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated to obtain the crude product. The crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 47%~95%, flow rate: 30 mL / min) to obtain the title compound 23 (10.03 mg, yield: 13.5%). MS m / z (ESI): 1102.2(M+1) + . 1H NMR (400 MHz, CDCl3) δ 8.94 (s, 1H), 8.18-8.14 (m, 2H), 7.65-7.53 (m, 2H), 7.43 (s, 1H), 7.25-7.19 (m, 6H), 6.66 (s, 2H), 6.59 (s, 1H), 5.75-5.63 (m, 3H), 5.32-5.13 (m, 3H), 4.51 (d, 2H), 4.08 (s, 4H), 3.89 (s, 3H), 3.75-3.61 (m, 2H), 3.48 (t, 2H), 3.42-3.31 (m, 1H), 3.08 (d, 2H), 2.38-2.11 (m, 7H), 1.89-1.85 (m, 2H), 1.30-1.25 (m, 7H), 1.20 -1.17 (m, 1H), 1.49-1.09 (m, 1H), 1.02 (t, 3H), 0.88-0.75 (m, 6H).

[0256] Example 24 (R)-N-((1R, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyran[3',4':6,7]indolidine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyran[3',4':6,7]indolidine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide [ka]

[0257] Step 1: N-(3-fluoro-4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborinan-2-yl)phenyl)acetamide 24b N-(3-fluoro-5-iodo-4-methylphenyl)acetamide 24a (7 g, 23.9 mmol) was dissolved in 1,4-dioxane (70 mL), and bis(pinacolato)diborone (12.14 g, 47.8 mmol), Pd(dppf)Cl2 (3.47 g, 4.78 mmol), and potassium acetate (7.04 g, 71.7 mmol) were added. The reaction was stirred at 110°C for 16 hours under nitrogen gas protection. After the reaction was complete, the mixture was filtered, the filtrate was collected, and the solvent was evaporated by rotation to obtain the crude product. The crude product was purified by column chromatography separation system B to obtain the title compound 24b (4.1 g, yield: 59%). MS m / z (ESI): 294.1 (M+1) + .

[0258] Step 2: N-(3-fluoro-5-hydroxy-4-methylphenyl)acetamide 24c Compound 24b (4.1 g, 14 mmol) was dissolved in a mixed solvent of tetrahydrofuran and water (40 mL, V / V=1:1), and sodium chlorite (1.52 g, 16.8 mmol) was added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, ethyl acetate was added, and the organic phase was washed three times with aqueous sodium hydroxide solution (15% wt). The aqueous phase was collected, the pH was adjusted to 3-5 with hydrochloric acid, and then extracted with dichloromethane to collect the organic phase. The solvent was evaporated by rotation to obtain the crude product. The crude product was purified by separation system B of silica gel column chromatography to obtain the title compound 24c (1.9 g, yield: 74%). MS m / z (ESI): 184.0 (M+1) + .

[0259] Step 3: 4-(5-acetamido-3-fluoro-2-methylphenoxy)methyl butyrate 24d Compound 24c (1.9 g, 10.4 mmol) was dissolved in N,N-dimethylformamide (20 mL), and methyl 4-bromobutyrate (2.82 g, 15.6 mmol), potassium carbonate (2.87 g, 20.8 mmol), and potassium iodide (2.59 g, 15.6 mmol) were added. The reaction was stirred at 85°C for 16 hours. After the reaction was complete, ethyl acetate was added, and the system was washed three times with saturated brine to collect the organic phase. The solvent was evaporated by rotation to obtain the crude product, which was purified by silica gel column chromatography separation system B to obtain the title compound 24d (2 g, yield: 68%). MS m / z (ESI): 284.0 (M+1) + .

[0260] Step 4 4-(5-acetamido-3-fluoro-2-methylphenoxy)butyrate 24e Compound 24d (2 g, 7.1 mmol) was dissolved in a mixed solvent of tetrahydrofuran and water (20 mL, V / V=1:1), and lithium hydroxide (340 mg, 14.2 mmol) was added to the system. The reaction was stirred at room temperature for 2 hours. After the reaction was complete, dilute hydrochloric acid was added to adjust the pH of the system to 3-5, ethyl acetate was added for extraction, and the organic phase was collected and dried over anhydrous sodium sulfate. The solvent was evaporated by rotation to obtain crude product 24e (1.4 g, yield: 73%), and the product was used directly in the next step without purification. MS m / z (ESI): 270.0 (M+1) + .

[0261] Step 5 N-(8-fluoro-9-methyl-5-oxo-2,3,4,5-tetrahydrobenzo[b]oxan-6-yl)acetamide 24f Compound 24e (1.4 g, 5.2 mmol) was dissolved in trifluoroacetic acid (20 mL), and trifluoroanhydride (3.28 g, 15.6 mmol) was added. The reaction was stirred at 40°C for 1 hour. After the reaction was complete, water (10 mL) was slowly added, and the mixture was stirred at 40°C for another 1 hour. After the reaction was complete, the system was extracted with ethyl acetate, and the organic phase was collected. The solvent was evaporated by rotation to obtain the crude product, which was purified by silica gel column chromatography separation system B to obtain compound 24f (900 mg, yield: 69%). MS m / z (ESI): 252.1 (M+1) + . 1 H NMR (400 MHz, CDCl3) δ 11.24 (s, 1H), 8.10 (d, 1H), 4.15 (t, 2H), 2.84 - 2.77 (m, 2H), 2.17 - 2.10 (m, 5H), 2.07 (d, 3H).

[0262] Step 6 (Z)-N-(8-fluoro-4-(hydroxyimino)-9-methyl-5-oxo-2,3,4,5-tetrahydrobenzo[b]oxan-6-yl)acetamide 24g Tetrahydrofuran (20 mL) and tert-butanol (5 mL) were added to a flask, the mixture was cooled to 0°C, and potassium tert-butoxide (806 mg, 6.43 mmol) was added. Then compound 24f (900 mg, 3.59 mmol) was slowly added, and the reaction was stirred at 0°C for 10 minutes. Isoamyl nitrite (631 mg, 5.39 mmol) was added, and the reaction was stirred at 0°C for 1 hour. After the reaction was complete, hydrochloric acid was added to adjust the pH to 4-5, and the mixture was extracted with ethyl acetate. The organic phase was washed with brine, dried over anhydrous sodium sulfate, concentrated, and the solvent was evaporated by rotation to obtain 24 g (900 mg) of crude product, which was used directly in the next step without purification. MS m / z (ESI): 281.0 (M+1) + .

[0263] Step 7 (9H-Fluoren-9-yl)methyl(6-acetamido-8-fluoro-9-methyl-5-oxo-2,3,4,5-tetrahydrobenzo[b]oxetan-4-yl)carbamate 24h 24 g (900 mg, 3.21 mmol) of the compound was dissolved in dioxane (10 mL), and palladium-carbon catalyst (200 mg) and 1 M dilute hydrochloric acid (2 mL) were added. The reaction was stirred at room temperature for 2 hours under a hydrogen gas environment. After the reaction was complete, the mixture was filtered, the filtrate was collected, and saturated sodium bicarbonate solution was added to adjust the pH to 10. Then, 929 mg (3.59 mmol) of chloroformate-9-fluorenylmethyl ester was added, and the reaction was stirred at room temperature for 1 hour. After the reaction was complete, the system was extracted with dichloromethane, the organic phase was collected, and the solvent was evaporated by rotation to obtain the crude product. The crude product was purified by silica gel column chromatography system B to obtain the title compound 24h (500 mg, yield: 29%). MS m / z (ESI): 489.1 (M+1) + .

[0264] Step 8 (9H-Fluoren-9-yl)methyl(6-acetamido-8-fluoro-9-methyl-5-oxo-2,3,4,5-tetrahydrobenzo[b]oxetan-4-yl)carbamate 24i Compound 24h (500 mg, 1.02 mmol) was dissolved in dichloromethane (5 mL), and a methanol solution of 4 M hydrogen chloride (5 mL) was added. The reaction was stirred at 60°C for 2 hours. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product. The crude product was separated and purified by silica gel column chromatography to obtain the title compound 24i (350 mg, yield: 77%). MS m / z (ESI): 447.1 (M+1) + .

[0265] Step 9 (9H-fluoro-9-yl)methyl((10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)carbamate 24j Compound 24i (350 mg, 0.78 mmol) was dissolved in toluene (5 mL), and (S)-4-ethyl-4-hydroxy-7,8-dihydro-1H-pyran[3,4-f]indolidine-3,6,10(4H)-trione (248 mg, 0.94 mmol) and p-toluenesulfonic acid monohydrate (223 mg, 1.18 mmol) were added. The reaction was stirred at 120°C for 2 hours. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product, which was purified by silica gel column chromatography separation system B to obtain the title compound 24j (450 mg, yield: 85%). MS m / z (ESI): 674.2 (M+1) + .

[0266] Step 10 (10S)-1-amino-10-ethyl-6-fluoro-10-hydroxy-5-methyl-1,2,3,10,13,16-hexahydro-11H,14H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-11,14-one 24k Compound 24j (100 mg, 0.15 mmol) was dissolved in N,N-dimethylformamide, and diethylamine (0.6 mL) was added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the solvent was evaporated by rotation to obtain crude product 24k (65 mg, yield: 97%). MS m / z (ESI): 452.1 (M+1) + .

[0267] Step 11 (R)-N-((1R, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyran[3',4':6,7]indolidine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide (R)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyran[3',4':6,7]indolidine[1,2-b]quinoline-1-yl)-3-hydroxybutanamide 24 Compound 24k (65 mg, 0.14 mmol) was dissolved in N,N-dimethylformamide (3 mL), and (R)-3-hydroxybutyric acid (18 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (82 mg, 0.22 mmol), and N,N-diisopropylethylamine (37 mg, 0.29 mmol) were added. The reaction was stirred at room temperature for 15 minutes. After the reaction was complete, the crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2, chromatography column: Xbridge C18 150 x 19 mm, 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 10 minutes, gradient ratio: acetonitrile phase 37%~47%, flow rate: 20 mL / min) to obtain the title compounds 24-1 (7.37 mg, yield: 10%) and 24-2 (4.70 mg, yield: 6%).

[0268] Single-stereoconfiguration compound 24-1 (short retention time) MS m / z (ESI): 538.2 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.48 (d, 1H), 7.68 (d, 1H), 7.47 (d, 1H), 7.29 (s, 1H), 7.11 (d, 1H), 5.61 (dd, 1H), 5.51 - 5.39 (m, 4H), 4.61 - 4.50 (m, 1H), 4.30 - 4.16 (m, 1H), 3.92 - 3.80 (m,1H), 2.35 (d, 3H), 2.29 (s, 1H), 2.20 (dd, 2H), 2.05 - 1.94 (m,1H), 1.92 - 1.77 (m, 2H), 0.87 (t, 3H), 0.82 (d, 3H).

[0269] Single-stereoconfiguration compounds 24-2 (long retention time) MS m / z (ESI): 538.2 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.48 (d, 1H), 7.68 (d, 1H), 7.47 (d, 1H), 7.29 (s, 1H), 7.11 (d, 1H), 5.68 (dd, 1H), 5.48 (d, 2H), 5.43 (s, 2H), 4.54 - 4.42 (m,1H), 4.33 - 4.24 (m, 1H), 3.92 - 3.81 (m,1H), 2.35 (d, 3H), 2.29 (s, 1H), 2.26 - 2.16 (m, 2H), 2.03 - 1.93 (m,1H), 1.92 - 1.78 (m, 2H), 1.00 (d, 3H), 0.87 (t, 3H).

[0270] Example 25 (1S,3S)-N-((1R,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide (1S,3R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide [ka]

[0271] Step 1 (1S,3S)-N-((1R,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide (1S,3R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxycyclobutan-1-carboxamide Compound 24k (65 mg, 0.14 mmol) was dissolved in N,N-dimethylformamide (3 mL), and cis-3-hydroxycyclobutylcarboxylic acid (20 mg, 0.17 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (82 mg, 0.22 mmol), and N,N-diisopropylethylamine (37 mg, 0.29 mmol) were added. The reaction system was stirred at room temperature for 15 minutes. After the reaction was complete, the crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge C18 150 x 19 mm, 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 18 minutes, gradient ratio: acetonitrile phase 34%~44%, flow rate: 20 mL / min) to obtain the title compounds 25-1 (6.57 mg, yield: 9%) and 25-2 (8.90 mg, yield: 12%).

[0272] Single-stereoconfiguration compound 25-1 (short retention time) MS m / z (ESI): 550.1 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.40 (d, 1H), 7.67 (d, 1H), 7.29 (s, 1H), 5.65 (dd, 1H), 5.50 (d, 1H), 5.44 (s, 2H), 5.29 (d, 1H), 4.45 - 4.37 (m, 1H), 4.36 - 4.25 (m, 1H), 3.95 - 3.82 (m, 1H), 2.57 - 2.53 (m, 1H), 2.48 - 2.44 (m, 2H), 2.35 (d, 3H), 2.30 - 2.24 (m, 1H), 2.22 - 2.15 (m, 1H), 2.06 - 1.69 (m, 6H), 0.86 (dd, 3H).

[0273] Single-stereoconfiguration compound 25-2 (long retention time) MS m / z (ESI): 550.1 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.41 (d, 1H), 7.68 (d, 1H), 7.29 (s, 1H), 5.67 (dd, 1H), 5.48 (d, 1H), 5.41 (d, 2H), 5.28 (d, 1H), 4.44 - 4.26 (m, 2H), 3.97 - 3.85 (m, 1H), 2.59 - 2.53 (m, 1H), 2.47 - 2.42 (m, 2H), 2.34 (t, 3H), 2.30-2.25 (m, 1H), 2.24 - 2.18 (m, 1H), 2.07 - 1.72 (m, 6H), 0.87 (t, 3H).

[0274] Example 26 N-((7S,15R)-7-benzyl-17-(((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexa-5-inamide [ka]

[0275] Step 1: 5-Bromo-2-(methylthio)nicotinonitrile 26b 5-Bromo-2-chloronicotinonitrile 26a (2 g, 9.2 mmol) was dissolved in ethylene glycol dimethyl ether (20 mL), and sodium methyl mercaptan (640 mg, 9.2 mmol) was added under ice bath. The reaction was stirred at room temperature for 4 hours. After the reaction was complete, aqueous ammonium chloride solution was added to quench the reaction, the system was extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, the organic phases were combined, and the mixture was concentrated. The resulting concentrate was purified by silica gel column chromatography system B to obtain the title compound 26b (1.5 g, yield: 71%). MS m / z (ESI): 228.9, 230.9 (M+1) + .

[0276] Step 2: 6-(5-cyano-6-(methylthio)pyridine-3-yl)hexyl-5-ic acid 26d 5-Bromo-2-(methylthio)nicotinonitrile 26b (1.5 g, 6.6 mmol) was dissolved in a mixed solvent of tetrahydrofuran (5 mL) and triethylamine (5 mL), and hexa-5-ic acid 26c (1.5 g, 13.2 mmol), bistriphenylphosphine palladium dichloride (913 mg, 1.3 mmol), and cuprous iodide (133 mg, 0.7 mmol) were added. The reaction was stirred at 70°C for 2 hours. After the reaction was complete, the reaction mixture was filtered and the mother liquor was stirred directly. The crude product obtained was purified by silica gel column chromatography system B to obtain 6-(5-cyano-6-(methylthio)pyridine-3-yl)hexyl-5-ic acid 26d (0.7 g, yield: 41%). MS m / z (ESI): 261.1 (M+1) + .

[0277] Step 3: 6-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexyl-5-ic acid 26e 6-(5-cyano-6-(methylthio)pyridine-3-yl)hexyl-5-ic acid 26d (0.7 g, 2.7 mmol) was dissolved in methanol (10 mL) and water (10 mL), and potassium peroxymonosulfate (9.3 g, 27 mmol) was added. The reaction was stirred at room temperature for 2 hours. After the reaction was complete, the mixture was filtered, the mother liquor was poured into water, extracted three times with dichloromethane, dried over anhydrous sodium sulfate, the organic phases were combined and concentrated. The filtrate was collected to obtain crude product 26e (815 mg), which was used directly in the next step without purification. MS m / z (ESI): 293.0 (M+1) + .

[0278] Step 4 2,5-Dioxopyrrolidone-1-yl-6-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexyl-5-ic acid ester 26f Crude product 26e (815 mg) was dissolved in dichloromethane (10 mL), and N-hydroxysuccinimide (345 mg, 3.0 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.04 g, 5.4 mmol) were added. The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was poured into water, extracted three times with dichloromethane, dried over sodium sulfate, and the organic phases were combined and concentrated. The resulting concentrate was purified by silica gel column chromatography system B to obtain the title compound 26f (400 mg, yield: 37%). MS m / z (ESI): 390.0 (M+1) + .

[0279] Step 5 (3R,11S)-11-benzyl-24-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)-3-methyl-7,10,13,16,19-pentaoxo-4-oxa-6,9,12,15,18-pentazatetracarbonyl-23-enoic acid 26g Compound 26f (200 mg, 0.5 mmol) was dissolved in N,N-dimethylformamide (5 mL), and compound 13j (249 mg, 0.6 mmol) and N,N-diisopropylethylamine (130 mg, 1 mmol) were added. The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge C18 150 x 19 mm, 5 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 30%~40%, flow rate: 20 mL / min) to obtain 26 g (80 mg, yield: 21%) of the title compound. MS m / z (ESI): 748.3 (M+1) + .

[0280] Step 6 N-((7S,15R)-7-benzyl-17-(((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexa-5-inamide 26 26 g (25 mg, 0.03 mmol) of the compound was dissolved in N,N-dimethylformamide (5 mL), and compound 6m-2 (15 mg, 0.03 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (15 mg, 0.04 mmol), and N,N-diisopropylethylamine (8 mg, 0.06 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 45%~55%, flow rate: 30 mL / min) to obtain the title compound 026 (20 mg, yield: 51.7%). MS m / z (ESI): 1157.3 (M+1) + . 1 H NMR (400 MHz, CD3OD) δ 8.81 (d, 1H), 8.41 (d, 1H), 7.54 (s, 1H), 7.45 (d, 1H), 7.17 (s, 5H), 5.65 (dd, 1H), 5.57 (d, 1H), 5.43 - 5.30 (m, 3H), 4.71 (dd, 2H), 4.50 (dd, 1H), 4.22 (t, 1H), 3.81 (s, 4H), 3.64 (dd, 2H), 3.39 (s, 3H), 3.31 (s, 3H), 3.11 (dd, 1H), 2.90 (dd, 1H), 2.64 - 2.55 (m, 3H), 2.45 (d, 7H), 2.17 (dd, 1H), 2.00 - 1.86 (m, 5H), 1.28 (d, 3H), 1.00 (s, 3H).

[0281] Example 27 N-((7S,15R)-7-benzyl-17-(((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(4-cyclopropyl-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide [ka]

[0282] Step 1: 5-Bromo-2-chloro-4-cyclopropylpyrimidine 27b 5-Bromo-2-chloropyrimidine 27a (8.50 g, 43.94 mmol) was added to water (50 mL), cyclopropylcarboxylic acid (4.54 g, 52.73 mmol), silver nitrate (1.49 g, 8.79 mmol), and trifluoroacetic acid (2.51 g, 21.97 mmol) were added, and the reaction was heated to 70°C and stirred. Ammonium persulfate (20.01 g, 87.89 mmol) was slowly added, and the reaction was continued with stirring for 2 hours. After the reaction was complete, the system was extracted with ethyl acetate, the organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated by rotation to obtain the crude product. The crude product was purified by separation system B of column chromatography to obtain the title compound 27b (6 g, yield: 58%). MS m / z (ESI): 232.9, 235.0, 237.0 (M+1) + .

[0283] Step 2: 6-(2-chloro-4-cyclopropylpyrimidine-5-yl)hexyl-5-ic acid 27c Compound 27b (3 g, 12.85 mmol) was dissolved in tetrahydrofuran (20 mL), and 5-hexic acid (2.16 g, 19.28 mmol), bis(triphenylphosphine)dichloropalladium (1.80 g, 2.57 mmol), cuprous iodide (246 mg, 1.29 mmol), and triethylamine (3.9 g, 38.55 mmol) were added. The reaction was carried out with stirring at 70°C for 2 hours under nitrogen gas protection. After the reaction was complete, the mixture was filtered, the filtrate was collected, and the solvent was evaporated by rotation to obtain the crude product. The crude product was subjected to separation and purification system B of column chromatography to obtain the title compound 27c (1.9 g, yield: 56%). MS m / z (ESI): 265.1, 267.1 (M+1) + .

[0284] Step 3: 6-(4-cyclopropyl-2-(methylthio)pyrimidine-5-yl)hexyl-5-ic acid 27d Compound 27c (600 mg, 2.27 mmol) was dissolved in N,N-dimethylformamide (5 mL), and sodium methyl mercaptan (191 mg, 2.72 mmol) and anhydrous magnesium sulfate (644 mg, 4.54 mmol) were added. The reaction was stirred at 50°C for 2 hours. Then, saturated aqueous ammonium chloride solution was added to the system, extracted with dichloromethane, and the organic phase was collected. The solvent was evaporated by rotation to obtain crude product 27d (600 mg). The crude product was used directly in the next step without purification. MS m / z (ESI): 277.0 (M+1) + .

[0285] Step 4 6-(4-cyclopropyl-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-ic acid 27e Compound 27d (600 mg, 2.17 mmol) was dissolved in a mixed solvent of methanol and water (V / V = 1:1, 10 mL), and potassium peroxymonosulfate (6.67 g, 10.86 mmol) was added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the mixture was filtered and the filtrate was collected. The filtrate was extracted with dichloromethane, the organic phase was collected, dried over anhydrous sodium sulfate, washed with saturated brine, and the solvent was evaporated by rotation to obtain the crude product. The crude product was purified by separation system B of column chromatography to obtain the title compound 27e (200 mg, yield: 30%). MS m / z (ESI): 309.1 (M+1) + .

[0286] Step 5: 2,5-Dioxopyrrolidone-1-yl-6-(4-cyclopropyl-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-ic acid ester 27f Compound 27e (200 mg, 0.65 mmol) was dissolved in dichloromethane, and N-hydroxysuccinimide (82 mg, 0.71 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (249 mg, 1.30 mmol) were added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product, which was purified by column chromatography separation system B to obtain compound 27f (200 mg, yield: 76%). MS m / z (ESI): 406.0 (M+1) + .

[0287] Step 6 (3R,11S)-11-benzyl-24-(4-cyclopropyl-2-(methylsulfonyl)pyrimidine-5-yl)-3-methyl-7,10,13,16,19-pentaoxo-4-oxa-6,9,12,15,18-pentazatetracarbonyl-23-enoic acid 27g Compound 27f (80 mg, 0.20 mmol) was dissolved in N,N-dimethylformamide (2 mL), and compound 13j (107 mg, 0.23 mmol) and N,N-diisopropylethylamine (51 mg, 0.39 mmol) were added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge C18 150 x 19 mm, 5 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 10 minutes, gradient ratio: acetonitrile phase 40%~50%, flow rate: 20 mL / min) to obtain 27 g (50 mg, yield: 34%) of the title compound. MS m / z (ESI): 764.2 (M+23) + .

[0288] Step 7 N-((7S,15R)-7-benzyl-17-(((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(4-cyclopropyl-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide 27 Compound 6m-2 (15 mg, 0.03 mmol) was dissolved in N,N-dimethylformamide (1 mL), and 27 g (25 mg, 0.03 mmol) of the compound, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (15 mg, 0.04 mmol) and N,N-diisopropylethylamine (9 mg, 0.07 mmol) were added. The reaction was stirred at room temperature for 15 minutes, and after the reaction was complete, the reaction mixture was separated and subjected to preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18). The compound was purified using a 21.2*250mm, 10μm mobile phase (1: water, containing 0.1% FA; 2: acetonitrile; gradient for 10 minutes, gradient ratio: acetonitrile phase 40%~70%, flow rate: 30 mL / min) to obtain the title compound 27 (18 mg, yield: 46%). MS m / z (ESI): 1173.3 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.86 (s, 1H), 8.67 (d, 1H), 8.49 (t, 1H), 8.29 (t, 1H), 8.20 (t, 1H), 8.12 (d, 1H), 8.06 (t, 1H), 7.73 (d, 1H), 7.30 (s, 1H), 7.23 - 7.17 (m, 4H), 7.17 - 7.11 (m,1H), 6.53 (s, 1H), 5.59 - 5.48 (m, 1H), 5.47 - 5.37 (m, 2H), 5.29 (s, 2H), 4.63 - 4.54 (m, 1H), 4.53 - 4.44 (m, 2H), 4.04 - 3.94 (m, 2H), 3.74 (d, 1H), 3.71 - 3.66 (m, 4H), 3.65 (d, 1H), 3.60 (d, 1H), 3.55 (d, 1H), 3.02 (dd, 2H), 2.86 - 2.72 (m, 3H), 2.41 (s, 3H), 2.28 - 2.22 (m, 1H), 2.05 - 1.97 (m, 1H), 1.92 -1.88 (m, 1H), 1.87 - 1.79 (m, 4H), 1.79 - 1.73 (m, 1H), 1.32 - 1.26 (m, 2H), 1.23 (s, 2H), 1.20 - 1.15 (m, 2H), 1.13 (d, 3H), 0.86 (t, 3H).

[0289] Example 28 N-((7S, 15R)-7-benzyl-17-(((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide [ka]

[0290] Step 1: 5-Bromo-2-chloro-4-methoxypyrimidine 28b 5-Bromo-2,4-dichloropyrimidine 28a (5.0 g, 22 mmol) was dissolved in methanol (60 mL), and a methanol solution of sodium methoxide (30%, 4.0 g, 22.0 mol) was added to the solution. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was concentrated, extracted under acetic acid inhibition, the organic phase was dried, and concentrated to obtain the title compound 28b (5.0 g, yield: 100%). MS m / z (ESI): 223.1 (M+1) + .

[0291] Step 2: 5-Bromo-4-methoxy-2-(methylthio)pyrimidine 28c Add methyl mercaptan sodium (1.52 g, 22 mmol) to N,N-dimethylformamide (25 mL) of compound 28b (5.0 g, 22.0 mmol), and mix the reaction mixture for 40 minutes. o The mixture was stirred in 1°C for 1 hour. After the reaction was complete, the reaction solution was poured into water, extracted with ethyl acetate (50 mL x 3), the organic phases were combined, dried, and concentrated. The resulting crude product was purified by silica gel column chromatography system B to obtain the title compound 28c (3.0 g, yield: 60%). MS m / z (ESI): 235.0 (M+1) + .

[0292] Step 3: 6-(4-methoxy-2-(methylthio)pyrimidine-5-yl)hexyl-5-ic acid 28d Compound 28c (3.0 g, 12.8 mmol) was dissolved in isopropanol (20 mL), and hexa-5-ic acid (1.43 g, 12.8 mmol), cuprous iodide (243 mg, 1.29 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (300 mg, 0.41 mmol), and sodium carbonate solution (5 M, 8 mL) were added. The reaction mixture was stirred at 80°C for 16 hours under a nitrogen atmosphere. After the reaction was complete, the reaction mixture was filtered, the filtrate was extracted with ethyl acetate (50 mL x 3), the organic phases were combined, dried, and concentrated to obtain the crude product, which was purified by silica gel column chromatography system B to obtain the title compound 28d (3.0 g, yield: 87%). MS m / z (ESI): 267.1 (M+1) + .

[0293] Step 4 5-bromo-2,4-dichloropyrimidine 6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-ic acid 28e Compound 28d (3.0 g, 11.2 mmol) was dissolved in a mixed solvent of methanol and water (40 mL, V / V=1:1), potassium sulfate peroxide (11.6 g, 33.6 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the reaction mixture was poured into water and extracted with ethyl acetate (50 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate to obtain the crude product. The crude product was purified by silica gel column chromatography system B to obtain the title compound 28e (2.5 g, yield: 75%). MS m / z (ESI): 299.1 (M+H) + .

[0294] Step 5 2,5-Dioxopyrrolidone-1-yl-6-(4-Methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-ic acid ester 28f Compound 28e (700 mg, 2.3 mmol) and N-hydroxysuccinimide (396 mg, 3.45 mmol) were dissolved in tetrahydrofuran (10 mL), to which N,N-diisopropylcarbodiimide (434 mg, 3.45 mmol) was added, and the reaction was stirred at room temperature for 3 hours. After the reaction was complete, the reaction mixture was directly concentrated, and the resulting crude product was purified by silica gel column chromatography system B to obtain the title compound 28f (600 mg, yield: 60%). MS m / z (ESI): 395.5 (M+1) + .

[0295] Step 6 (3R, 11S)-11-benzyl-24-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)-3-methyl-7,10,13,16,19-pentaoxo-4-oxa-6,9,12,15,18-pentazatetracarbonyl-23-enoic acid 28g Compound 28f (600 mg, 1.5 mmol) was dissolved in N,N-dimethylformamide (6 mL), and N,N-diisopropylethylamine (580 mg, 4.5 mmol) and compound 13j (677 mg, 1.5 mmol) were sequentially added to the solution. The reaction mixture was stirred at room temperature for 3 hours. The mixture was concentrated to obtain the crude product, which was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: Xbridge C18 150 x 19 mm, 5 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 29%~100%, flow rate: 25 mL / min) to obtain 28 g (350 mg, yield: 35%) of the title compound. MS m / z (ESI): 754.1 (M+Na) + .

[0296] Step 7 N-((7S, 15R)-7-benzyl-17-(((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide 28 28 g (25 mg, 0.034 mmol) of the compound was dissolved in N,N-dimethylformamide (3 mL), and compound 6m-2 (15 mg, 0.033 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (25 mg, 0.065 mmol), and N,N-diisopropylethylamine (171 mg, 1.32 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the mixture was concentrated to obtain the crude product, which was then purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 40%~70%, flow rate: 30 mL / min) to obtain the title compound 28 (16 mg, yield: 42%). MS m / z (ESI): 1163.2 (M+1) + . 1H NMR (400 MHz, CDCl3) δ 8.56 (s, 1H), 8.43 (s, 1H), 7.52 - 7.48 (m, 2H), 7.40 - 7.29 (m, 3H), 7.24 - 7.04 (m, 6H), 6.94 (s, 1H), 5.60 - 5.55 (m, 2H), 5.35 - 5.32 (m, 1H), 5.16 (d, 2H), 5.04 - 4.56 (m, 4H), 4.25 -3.73 (m, 9H), 3.30 (s, 3H), 3.26 - 2.93 (m, 4H), 2.60 - 2.42 (m, 7H), 2.26 (s, 3H), 2.17 - 1.74 (m, 8H), 1.31 (d, 3H), 0.97 (t, 3H).

[0297] Example 29 (R)-3-(((S)-7-benzyl-17-(4-(2-(methylsulfonyl)pyrimidine-5-yl)piperidine-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentazaheptadecyl)oxy)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,3,4,10,11,14,16-octahydro-13H-cycloheptane[de]pyrano[3',4':6,7]indolidino[1,b]quinoline-1-yl)butanamide [ka]

[0298] Step 1: 4-Iodopiperidine hydrochloride 29b 4-iodopiperidine-1-carboxylate tert-butyl 29a (4 g, 12.86 mmol) was added to the reaction flask, and a 1,4-dioxane solution in 4 M hydrochloric acid (20 ml) was added at room temperature. The reaction was allowed to proceed at room temperature for 1 hour. After the reaction was complete, the solvent system was dried by rotation to obtain the crude product 4-iodopiperidine hydrochloride 29b (2.7 g, yield: 100%). MS m / z (ESI): 212.0 (M+1) + .

[0299] Step 2: Ethyl 3-(4-iodopiperidine-1-yl)propionate 29c 4-iodopiperidine hydrochloride 29b (2.7 g, 10.93 mmol), ethyl 3-bromopropionate (2.95 g, 16.40 mmol), and potassium carbonate (7.54 g, 54.65 mmol) were added to the reaction flask, and acetonitrile (50 ml) was added. The reaction was carried out at 60°C for 16 hours. After the reaction was complete, the solvent system was evaporated by rotation, methyl tert-butyl ether (50 mL) was added, and the mixture was beaten for 30 minutes. The filtrate was filtered, and the mixture was evaporated by rotation to obtain crude product 29c (3.1 g). The product was used directly in the next step without purification. MS m / z (ESI): 312.1 (M+1) + .

[0300] Step 3: 3-(4-(2-(methylthio)pyrimidine-5-yl)piperidine-1-yl)propionate ethyl 29d Nickel dimethoxyethane chloride (299 mg, 1.37 mmol) and 2-carboxyimidoamide hydrochloride (215 mg, 1.37 mmol) were added to a reaction flask, and dimethylacetamide (10 ml) was added under a nitrogen atmosphere. The mixture was stirred at room temperature for 30 minutes. Another reaction flask was taken, and compound 29c (1.7 g, 5.46 mmol), 5-bromo-2-(methylthio)pyrimidine (1.1 g, 5.46 mmol), manganese (75 mg, 1.37 mmol), and tetrabutylammonium iodide (2.0 g, 5.46 mmol) were added. Then N,N-dimethylacetamide (10 ml) was added, and the mixture was substituted with nitrogen gas three times. After substitution was complete, the nickel ligand compound was transferred to the system using a syringe, and the reaction was carried out at 60°C for 3 hours. After the reaction was complete, ethyl acetate (200 ml) and water (400 ml) were added to the system for extraction. The organic phase was dried, concentrated, and purified by silica gel column chromatography system B to obtain the title compound 29d (1.5 g, yield: 89%). MS m / z (ESI): 310.2 (M+1) + .

[0301] Step 4 3-(4-(2-(methylthio)pyrimidine-5-yl)piperidine-1-yl)propionic acid 29e Compound 29d (1.5 g, 4.84 mmol) was added to a mixed solvent of tetrahydrofuran and water (15 mL, V / V = 2:1), and lithium hydroxide (348 mg, 14.52 mmol) was added. The mixture was reacted at room temperature for 16 hours. After the reaction was complete, the system was filtered, concentrated, and evaporated by rotation to obtain crude product 29e (1.5 g). The product was used directly in the next step without purification. MS m / z (ESI): 282.2 (M+1) + .

[0302] Step 5 3-(4-(2-(methylsulfonyl)pyrimidine-5-yl)piperidine-1-yl)propionic acid 29f Compound 29e (1.1 g) was dissolved in trifluoroacetic acid, and an excess amount of metachloroperbenzoic acid was added until the reaction was complete at room temperature. After the reaction was complete, the trifluoroacetic acid in the system was blown dry with nitrogen gas, and then water (30 ml) was added and beaten. The mixture was filtered, the filtrate was collected and concentrated, and purified by column chromatography system A to obtain compound 29f (430 mg). MS m / z (ESI): 314.1 (M+1) + .

[0303] Step 6 (3R, 11S)-11-benzyl-3-methyl-21-(4-(2-(methylsulfonyl)pyrimidine-5-yl)piperidine-1-yl)-7,10,13,16,19-pentaoxo-4-oxa-6,9,12,15,18-pentazaeicosanoic acid 29h Compound 29f (430 mg, 1.37 mmol) and N-hydroxysuccinimide (189 mg, 1.64 mmol) were dissolved in super-dehydrated N,N-dimethylformamide (10 ml), and then N,N-diisopropylcarbodiimide (206 mg, 1.64 mmol) was added, and the reaction was stirred at room temperature for 2 hours. Compound 13j (300 mg, 0.66 mmol) and N,N-diisopropylethylamine (213 mg, 1.65 mmol) were added using a one-pot synthesis method. After the reaction was complete, the crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 40%~95%, flow rate: 30 mL / min) to obtain the title compound 29h (200 mg). MS m / z (ESI): 747.3 (M+1) + .

[0304] Step 7 (R)-3-(((S)-7-benzyl-17-(4-(2-(methylsulfonyl)pyrimidine-5-yl)piperidine-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentazaheptadecyl)oxy)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,3,4,10,11,14,16-octahydro-13H-cycloheptane[de]pyrano[3',4':6,7]indolidino[1,b]quinoline-1-yl)butanamide Compound 29h (25 mg, 0.033 mmol), Compound 6m-2 (15 mg, 0.033 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (19 mg, 0.0495 mmol), and N,N-diisopropylethylamine (10.6 mg, 0.0825 mmol) were added to super-dehydrated N,N-dimethylformamide (5 ml). The reaction was carried out at room temperature for 2 hours. After the reaction was complete, the final product was subjected to preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10). The compound was purified using a μm mobile phase (water containing 0.1% FA, mobile phase 2: acetonitrile) with a gradient for 15 minutes (gradient ratio: acetonitrile phase 40% to 95%, flow rate: 30 mL / min) to obtain title compound 29 (16.7 mg, yield: 42.3%). MS m / z (ESI): 1178.4 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 9.00 (s, 2H), 8.66 (d, 1H), 8.48 (t, 1H), 8.35 (t, 1H), 8.27 (t, 1H), 8.11(d, 1H), 8.02 (t, 1H), 7.74 (d, 1H), 7.30 (s, 1H), 7.21 (d, 4H), 7.18 - 7.12 (m, 1H), 6.51 (s, 1H), 5.53 (s, 1H), 5.42(s, 2H), 5.29 (s, 2H), 4.58 (dd, 1H), 4.49 (dd, 2H), 3.99 (dd, 1H), 3.78 - 3.58 (m, 6H), 3.39 (s, 3H), 3.00 (t, 3H), 2.78 - 2.66 (m, 2H), 2.57 (t, 3H), 2.43 (t, 4H), 2.38 - 2.21 (m, 5H), 2.02 (t, 3H), 1.82 (ddd, 8H), 1.13 (d, 3H), 0.86 (t, 3H).

[0305] Example 30 N-((7S,15R)-7-benzyl-17-(((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-inamide [ka]

[0306] Step 1 N-((7S, 15R)-7-benzyl-17-(((1S, 9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-inamide 30 Compound 21b (30 mg, 0.065 mmol) was dissolved in anhydrous N,N-dimethylformamide (5 mL), and 28 g of compound (43 mg, 0.064 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (37 mg, 0.096 mmol), and Japanese N,N-diisopropylethylamine (171 mg, 1.32 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, water (50 mL) was added to the reaction mixture, extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated brine, dried, concentrated, and the crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 um; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 42%~52%, flow rate: 30 mL / min) to obtain the title compound 30 (18.5 mg, 23.5%). MS m / z (ESI): 1175.3 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1H), 8.53 - 8.45 (m, 2H), 8.30 (t, 1H), 8.17 (t, 1H), 8.12 (d, 1H), 8.05 (t, 1H), 7.74 (d, 1H), 7.30 (s, 1H), 7.27 - 7.14 (m, 5H), 6.52 (s, 1H), 5.59 - 5.55(m,1H), 5.42 (s, 2H), 5.20 (dd, 2H), 4.55 - 4.51 (m, 3H), 4.09 - 3.97 (m, 4H), 3.78 - 3.54 (m, 6H), 3.39 (s, 3H), 3.31 (s, 3H), 3.04 (dd, 1H), 2.80 - 2.75 (m, 1H), 2.56 - 2.53 (m, 1H), 2.46 - 2.42 (m, 1H), 2.35 - 2.20 (m, 4H), 2.16 - 2.12 (m, 2H), 1.94-1.78 (m, 4H), 1.15-1.12 (m, 5H), 0.87 (t, 3H), 0.79 - 0.72 (m, 2H).

[0307] Example 31 N-((7S,15R)-7-benzyl-17-(((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexyl-5-inamide [ka]

[0308] Step 1 N-((7S, 15R)-7-benzyl-17-(((1S, 9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexyl-5-inamide 31 26 g (50 mg, 0.06 mmol) of compound was dissolved in N,N-dimethylformamide (5 mL), and compound 21b (30 mg, 0.06 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (30 mg, 0.08 mmol), and N,N-diisopropylethylamine (16 mg, 0.12 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction mixture was separated and purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 42%~52%, flow rate: 20 mL / min) to obtain the title compound 31 (28 mg, yield: 40%). MS m / z (ESI): 1169.3 (M+1) + . 1H NMR (400 MHz, CD3OD) δ 8.77 (d, 1H), 8.37 (d, 1H), 7.57 (s, 1H), 7.51 (d, 1H), 7.23 - 7.19 (m, 2H), 7.15 (s, 3H), 5.58 (dd, 2H), 5.38 - 5.25 (m, 2H), 5.07 (d, 1H), 4.69 (d, 2H), 4.46 - 4.39 (m, 1H), 4.11 (dd, 1H), 3.85 (s, 5H), 3.59 (dd, 1H), 3.51-3.45 (m, 1H), 3.37 (s, 3H), 2.99 (ddd, 2H), 2.57 (t, 2H), 2.46 (dt, 4H), 2.34 - 2.22 (m, 2H), 2.01 - 1.81 (m, 6H), 1.26 (d, 3H), 1.13 (d, 2H), 0.97 (s, 3H), 0.80 (s, 2H).

[0309] Example 32 N-((7S,15R)-7-benzyl-17-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-amide [ka]

[0310] Step 1 N-((7S,15R)-7-benzyl-17-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-4-vinyl-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-15-methyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-amide 32 Compound 7b-2 (20 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 28 g (32 mg, 0.04 mmol) of the compound, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol), and N,N-diisopropylethylamine (11 mg, 0.09 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5 μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 12 minutes, gradient ratio: acetonitrile phase 42%~52%, flow rate: 20 mL / min) to obtain the title compound 32 (24 mg, yield: 46%). MS m / z (ESI): 1161.4 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1H), 8.52 (d, 1H), 8.31 (s, 1H), 8.16 (s, 1H), 8.10 (d, 1H), 8.04 (s, 1H), 7.85 (d, 1H), 7.34 (s, 1H), 7.25 - 7.15 (m, 4H), 6.97 (dd, 2H), 6.54 (s, 1H), 5.87 - 5.80 (m, 2H), 5.59 (s, 1H), 5.43 (s, 1H), 5.32 (d, 2H), 5.18 (d, 2H), 4.65 (s, 2H), 4.52 (d, 4H), 4.06 (s, 3H), 3.69 (d, 3H), 3.03 (d, 2H), 2.71 - 2.65 (m, 4H), 2.32 (dd, 4H), 2.11 (s, 3H), 1.91-1.75 (m, 6H), 1.12 (d, 3H), 0.90 - 0.82 (m, 3H).

[0311] Example 33 (R)-2-Cyclopropyl-N-((1S, 10S)-5-Cyclopropyl-10-ethyl-6-Fluoro-10-hydroxy-11,14-Dioxo-1,2,3,4,10,11,14,16-Octahydro-13H-Cyclohepta[3',4':6,7]Indolidino[1,2-b]Quinoline-1-yl)-2-Hydroxyacetamide (S)-2-Cyclopropyl-N-((1S, 10S)-5-Cyclopropyl-10-ethyl-6-Fluoro-10-hydroxy-11,14-Dioxo-1,2,3,4,10,11,14,16-Octahydro-13H-Cyclohepta[3',4':6,7]Indolidino[1,2-b]Quinoline-1-yl)-2-Hydroxyacetamide [ka]

[0312] Step 1 (R)-2-Cyclopropyl-N-((1S, 10S)-5-Cyclopropyl-10-ethyl-6-Fluoro-10-hydroxy-11,14-Dioxo-1,2,3,4,10,11,14,16-Octahydro-13H-Cyclohepta[3',4':6,7]Indolidino[1,2-b]Quinoline-1-yl)-2-Hydroxyacetamide (S)-2-Cyclopropyl-N-((1S, 10S)-5-Cyclopropyl-10-ethyl-6-Fluoro-10-hydroxy-11,14-Dioxo-1,2,3,4,11,14,16-Octahydro-13H-Cyclohepta[3',4':6,7]Indolidino[1,2-b]Quinoline-1-yl)-2-Hydroxyacetamide Compound 23b (20 mg, 0.042 mmol) was dissolved in N,N-dimethylacetamide (2 mL), and 2-cyclopropyl-2-hydroxyacetic acid (5.4 mg, 0.046 mmol), benzotriazole-1-yloxytri(dimethylamino)phosphonium hexafluorophosphate (22 mg, 0.051 mmol), and N,N-diisopropylethylamine (11 mg, 0.084 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 46%~100%, flow rate: 30 mL / min) to obtain products 33-1 (4.1 mg, yield: 17.2%) and 33-2 (1.7 mg, yield: 6.9%).

[0313] 33-1 (Single-stereoconfiguration compounds, short retention time) MS m / z (ESI): 574.2 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.62-8.37 (m, 2H), 7.69 (d, 1H), 7.28 (s, 1H), 6.53 (s, 1H), 5.59-5.48 (m, 2H), 5.43 (d, 2H), 5.28 (d, 1H), 3.62 (d, 1H), 3.49-3.46 (m, 1H), 2.19-1.72 (m, 6H), 1.69-1.53 ​​(m, 1H), 1.23 (s, 1H), 1.18-1.00 (m, 3H), 0.85 (t, 3H), 0.83-0.77 (m, 1H), 0.74-0.62 (m, 1H), 0.46-0.28 (m, 4H).

[0314] 33-2 (Single-stereoconfiguration compounds, long retention time) MS m / z (ESI): 574.2 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.45 (d, 2H), 7.69 (d, 1H), 7.28 (s, 1H), 6.53 (s, 1H), 5.60-5.52 (m, 1H), 5.48 (s, 1H), 5.43 (d, 2H), 5.30 (d, 1H), 3.59 (d, 1H), 3.49-3.46 (m, 1H), 2.14-1.79 (m, 6H), 1.75-1.61 (m, 1H), 1.24 (s, 1H), 1.17-1.02 (m, 3H), 0.86 (t, J = 8 Hz, 3H), 0.83-0.77 (m, 1H), 0.74-0.64 (m, 1H), 0.48-0.29 (m, 4H).

[0315] Example 34 (S)-3-Cyclopropyl-N-((1S,9S)-4-Cyclopropyl-9-ethyl-5-Fluoro-9-Hydroxy-10,13-Dioxy-2,3,9,10,13,15-Hexahydro-1H,12H-Benzo[d]pyrano[3',4':6,7]Indolidino[1,2-b]Quinoline-1-yl)-3-Hydroxypropanamide (R)-3-Cyclopropyl-N-((1S,9S)-4-Cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypropanamide [ka]

[0316] Step 1 (S)-3-Cyclopropyl-N-((1S,9S)-4-Cyclopropyl-9-ethyl-5-Fluoro-9-Hydroxy-10,13-Dioxy-2,3,9,10,13,15-Hexahydro-1H,12H-Benzo[d]pyrano[3',4':6,7]Indolidino[1,2-b]Quinoline-1-yl)-3-Hydroxypropanamide (R)-3-Cyclopropyl-N-((1S,9S)-4-Cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxy-2,3,9,10,13,15-Hexahydro-1H,12H-Benzo[d]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypropanamide Compound 21b (30 mg, 0.065 mmol) was dissolved in N,N-dimethylformamide (3 mL), and 3-cyclopropyl-3-hydroxypropionic acid (15 mg, 0.098 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (37 mg, 0.096 mmol), and N,N-diisopropylethylamine (171 mg, 1.32 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 40%~100%, flow rate: 30 mL / min) to obtain products 34-1 (4.2 mg, yield: 11%) and 34-2 (3.9 mg, yield: 10%).

[0317] 34-1 (Single-stereoconfiguration compounds, compounds with short retention times) MS m / z (ESI): 574.2 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.53-8.39 (m, 2H), 7.75 (d, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.55 (dd, 1H), 5.42 (s, 2H), 5.23 (s, 2H), 4.69 (s, 1H), 2.37 (d, 2H), 2.15 (d, 2H), 2.01-1.77 (m, 4H), 1.17-1.07 (m, 2H), 0.91-0.72 (m, 6H), 0.39-0.12 (m, 5H).

[0318] 34-2 (Single-stereoconfiguration compounds, compounds with long retention times) MS m / z (ESI): 574.2 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.45 (d, 2H), 7.75 (d, 1H), 7.31 (s, 1H), 6.53 (s, 1H), 5.64-5.54 (m, 1H), 5.43 (s, 2H), 5.24 (q, 2H), 4.65 (d, 1H), 2.33 (d, 2H), 2.27-1.73 (m, 6H), 1.12 (dd, 2H), 0.91-0.70 (m, 6H), 0.39-0.13 (m, 5H).

[0319] Example 35 (R)-2-Cyclopropyl-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide (S)-2-Cyclopropyl-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide [ka]

[0320] Step 1 (9H-fluoro-9-yl)methyl((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyran[3',4':6,7]indolidine[1,2-b]quinoline-1-yl)carbamate 035a Compound 024j (500 mg, 0.74 mmol) was purified by separating chiral isomers using a Gilson preparative and a Daicel chiral column (chromatography column: CHIRALPAK IE 3.0 cm ID × 25 cm, 10 μm; mobile phase 1: MeOH; mobile phase 2: DCM; gradient mixing ratio: MeOH / DCM = 80 / 20, flow rate: 25 mL / min) to obtain compound 035a (230 mg, yield: 46%). MS m / z (ESI): 674.1 (M+1) + .

[0321] Step 2 (1S, 10S)-1-amino-10-ethyl-6-fluoro-10-hydroxy-5-methyl-1,2,3,10,13,16-hexahydro-11H,14H-oxepino[4,3,2-de]pyran[3',4':6,7]indolidino[1,2-b]quinoline-11,14-one 035b Compound 035a (50 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (5 mL), and diethylamine (0.5 mL) was added. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the solvent was evaporated under vacuum by rotation to obtain the title compound 035b (34 mg), which was used directly in the next step without purification. MS m / z (ESI): 452.1 (M+1) + .

[0322] Step 3 (R)-2-Cyclopropyl-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide (S)-2-Cyclopropyl-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide Compound 035b (35 mg, 0.08 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 2-cyclopropyl-2-hydroxyacetic acid (11 mg, 0.09 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (46 mg, 0.12 mmol), and N,N-diisopropylethylamine (21 mg, 0.16 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5 μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 40%~60%, flow rate: 20 mL / min) to obtain 2-cyclopropyl-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-2-hydroxyacetamide (15 mg). The sample was chiral-resolved (SFC 80 chromatography column: Daicel CHIRALCEL OD, 250 mm x 30 mm ID, 10 μm; mobile phase: CO2 / MeOH [0.2% NH3 (7M Solution in MeOH)] = 70 / 30, flow rate: 70 g / min) to obtain compounds 35-1 (5 mg, yield: 12%) and 35-2 (5 mg, yield: 12%).

[0323] 35-1 (Single-stereoconfiguration compounds, compounds with short retention times) MS m / z (ESI): 550.1 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.14 (d, 1H), 7.72 (d, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.68-5.61 (m, 1H), 5.55-5.52 (m, 2H), 5.44 (s, 2H), 4.67 (dd, 1H), 4.23 (dd, 1H), 3.46 (t, 1H), 2.36 (s, 3H), 2.02-1.97 (m, 2H), 1.92-1.80 (m, 2H), 0.87 (t, 3H), 0.84 (d, 1H), 0.36-0.25 (m, 4H).

[0324] 35-2 (Single-stereoconfiguration compounds, compounds with long retention times) MS m / z (ESI): 550.1 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.17 (d, 1H), 7.71 (d, 1H), 7.30 (s, 1H), 6.53 (s, 1H), 5.62 (d, 1H), 5.49 (t, 2H), 5.44 (s, 2H), 4.62-4.53 (m, 1H), 4.29 (dd, 1H), 3.61 (t, 1H), 2.36 (d, 3H), 2.06-1.94 (m, 2H), 1.85 (dd, 2H), 0.87 (t, 3H), 0.85-0.83 (m, 1H), 0.25- 0.19 (m, 2H), 0.08 - 0.02 (m, 2H).

[0325] Example 36 (R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxy-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypentanamide (S)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxy-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypentanamide [ka]

[0326] Step 1 (R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxy-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypentanamide (S)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxy-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypentanamide Compound 6m-2 (30 mg, 0.07 mmol) was added to N,N-dimethylformamide (3 mL), and 3-hydroxypentanoic acid (9 mg, 0.07 mmol), N,N-diisopropylethylamine (17 mg, 0.13 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (38 mg, 0.10 mmol) were added. The reaction system was allowed to react at room temperature for 1 hour. After the reaction was complete, the compounds were purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, column: Xbridge 10μm C18 250 x 30mm, 10μm; mobile phase 1: water (containing 10 mmol / L FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile 5%~57%, flow rate: 50 mL / min) to obtain compounds 36-1 (6.98 mg, yield: 18%) and 36-2 (6.70 mg, yield: 18%).

[0327] 36-1 (Single steric configuration compound, short retention time compound) MS m / z (ESI): 550.2 (M+1) + . 1 H NMR (400MHz, DMSO-d6) δ 8.61 (d, 1H), 7.74 (d, 1H), 7.29 (s, 1H), 6.52 (s, 1H), 5.53 (s, 1H), 5.43 (s, 2H), 5.39-5.09 (m, 2H), 4.63 (s,1H), 2.43 (s, 3H), 2.32 (d, 3H), 2.10-1.93 (m, 3H), 1.92-1.81 (m, 2H), 1.75 (s, 1H), 1.40-1.32 (m, 2H), 1.24 (s, 2H), 0.91-0.76 (m, 6H).

[0328] 36-2 (compounds with a single three-dimensional configuration and compounds with long retention times) MS m / z (ESI): 550.2 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.69 (d, 1H), 7.73 (d, 1H), 7.28 (s, 1H), 6.51 (s, 1H), 5.51 (d, 2H), 5.43 (s, 2H), 5.33 (d, 1H), 4.64 (s, 1H), 2.42 (s, 4H), 2.34-2.22 (m, 4H), 2.08 (d, 1H), 2.01 (d, 1H), 1.92-1.78 (m, 2H), 1.65 (s, 1H), 1.44-1.31 (m, 2H), 1.24 (s, 1H), 0.95-0.66 (m, 6H).

[0329] Example 37 (S)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxy-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxy-4-methylpentanamide (R)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxy-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxy-4-methylpentanamide [ka]

[0330] Step 3 (S)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxy-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxy-4-methylpentanamide (R)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxy-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxy-4-methylpentanamide Compound 6m-2 (30 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (3 mL), and 3-hydroxy-4-methylpentanoic acid (9 mg, 0.07 mmol), N,N-diisopropylethylamine (17 mg, 0.13 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (38 mg, 0.10 mmol) were added. The reaction system was allowed to react at room temperature for 1 hour. After the reaction was complete, preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5 μm C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 42%~100%, flow rate: 20 mL / min) was performed. The compounds were purified to obtain compounds 37-1 (6.6 mg, yield: 18%) and 37-2 (6.9 mg, yield: 18%).

[0331] 37-1 (Single-stereoconfiguration compounds, compounds with short retention times) MS m / z (ESI): 564.2 (M+1) + . 1 H NMR (400MHz, DMSO-d6) δ 8.68 (d, 1H), 8.48 (br, 1H), 7.73 (d, 1H), 7.28 (s, 1H), 6.56 (s, 1H), 5.60-5.49 (m, 1H), 5.42 (s, 2H), 5.35 (d, 1H), 5.21 (d, 1H), 4.64 (s, 1H), 2.42 (s, 3H), 2.29 (d, 3H), 2.13-1.91 (m, 3H), 1.90-1.80 (m, 2H), 1.79-1.67 (m,1H),1.60-1.47 (m, 1H), 1.23 (s, 1H), 0.87 (d, 3H), 0.82 (d, 6H).

[0332] 37-2 (Single-stereoconfiguration compounds, compounds with long retention times) MS m / z (ESI): 564.2 (M+1)+ . 1 H NMR (400 MHz, DMSO-d6) δ 8.78 (s, 1H), 8.50 (s, 1H), 7.73 (s, 1H), 7.28 (s, 1H), 6.56 (s, 1H), 5.43 (s, 3H), 5.34 (d, 2H), 4.62 (s, 2H), 2.29 (s, 3H), 2.07 (s, 2H), 2.02-1.97 (m, 2H), 1.92-1.77 (m, 2H), 1.71-1.49 (m, 1H), 1.23 (s, 1H), 0.99-0.66 (m, 11H).

[0333] Example 38 (R)-3-Cyclopropyl-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypropanamide (S)-3-Cyclopropyl-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypropanamide [ka] (R)-3-Cyclopropyl-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypropanamide (S)-3-Cyclopropyl-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxypropanamide Compound 6m-2 (30 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (3 mL), and 3-cyclopropyl-3-hydroxypropionic acid (18 mg, 0.14 mmol), N,N-diisopropylethylamine (17 mg, 0.13 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (38 mg, 0.10 mmol) were added. The reaction system was allowed to react at room temperature for 1 hour. After the reaction was complete, the compounds were purified and separated by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: Xbridge 5μm C18 150 x 19mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 45%~100%, flow rate: 25 mL / min) to obtain the title compound 38-1 (4 mg, yield: 11%) and the title compound 38-2 (4 mg, yield: 11%).

[0334] 38-1 (Single-stereoconfiguration compounds, compounds with short retention times) MS m / z (ESI): 562.3 (M+1) + . 1H NMR (400MHz, DMSO-d6) δ 8.63 (d, 1H), 7.74 (d, 1H), 7.28 (s, 1H), 6.52 (s, 1H), 5.53 (s, 1H), 5.42 (s, 4H), 4.69 (d, 1H), 3.22 (s, 2H), 2.61 (d, 1H), 2.42 (s, 3H), 2.31 (d, 1H), 2.08-1.69 (m, 6H), 0.87 (s, 3H), 0.81 (d, 1H), 0.37-0.20 (m, 3H), 0.16-0.08 (m, 1H).

[0335] 38-2 (Single-stereoconfiguration compounds, compounds with long retention times) MS m / z (ESI): 562.3 (M+1) + . 1 H NMR (400 MHz, DMSO-d6) δ 8.71 (d, 1H), 7.73 (d, 1H), 7.27 (s, 1H), 6.51 (s, 1H), 5.43 (s, 5H), 4.64 (d, 1H), 4.55-4.51(m, 1H), 3.22 (s, 2H), 2.65 (d, 1H), 2.41 (s, 3H), 2.31 (d, 1H), 2.13-1.57 (m, 6H), 0.88 (s, 3H), 0.82 (s, 1H), 0.45 (s, 1H), 0.32-0.13 (m, 3H).

[0336] Example 39 (2S, 3R)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3', 4':6, 7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxy-2-methylbutanamide 39 [ka]

[0337] Step 1 (2S, 3R)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3', 4':6, 7]indolidino[1,2-b]quinoline-1-yl)-3-hydroxy-2-methylbutanamide 39 Compound 6m-2 (30 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (2 mL), and (2S, 3R)-3-hydroxy-2-methylbutyrate (12 mg, 0.10 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (38 mg, 0.10 mmol) were added. The reaction was stirred at room temperature for 15 minutes. After the reaction was complete, the compound was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: Xbridge 5μm C18 150 x 19mm, 5μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 10 minutes, gradient ratio: acetonitrile phase 42%~52%, flow rate: 25 mL / min) to obtain compound 39 (2 mg, yield: 5%). MS m / z (ESI): 550.0 (M+1) + . 1 H NMR (400 MHz, CDCl3) δ 7.73 (s, 1H), 7.42-7.28 (m, 2H), 5.51 (s, 1H), 5.41 (d, 1H), 5.35 (t, 1H), 5.26-5.16 (m, 2H), 5.04 (d, 1H), 4.18-4.00 (m, 2H), 3.33 (s, 2H), 2.65-2.50 (m, 3H), 2.48 (s, 3H), 2.27-2.18 (m, 1H), 2.11-2.04 (m, 2H), 2.03-1.97 (m, 1H), 1.33-1.28 (m, 6H), 0.95 (t, 3H).

[0338] Example 40 N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[yl]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxy-5,8,11,14-tetraazahexadecane-16-yl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-amide [ka]

[0339] Step 1: Preparation of 2-cyclopropyl-2-hydroxyacetic acid (40b) (S)-2-amino-2-cyclopropylacetic acid (23 g, 200 mmol) 40a was dissolved in 2 M sulfuric acid (100 mL), and 4 M aqueous sodium nitrite solution (450 mL) was added under ice bath. The reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction mixture was diluted with water, washed with ethyl acetate, and the resulting filtrate was directly concentrated to obtain crude product 40b (11.5 g). MS m / z (ESI): 117.1 (M+H) + .

[0340] Step 2: Preparation of 2-cyclopropyl-2-hydroxyacetate benzyl ester (40c) Compound 40b (11.5 g, 0.099 mol) was dissolved in acetonitrile (50 mL), and potassium carbonate (54.65 g, 0.396 mol), benzyl bromide (17.03 g, 0.099 mol), and tetrabutylammonium iodide (3.66 g, 0.0099 mol) were added sequentially. The reaction mixture was stirred at room temperature for 48 hours. The reaction mixture was filtered through diatomaceous earth, the filter cake was rinsed with ethyl acetate, and the filtrate was purified using silica gel column system B to obtain the title compound 40c (5.8 g, yield: 28%). MS m / z (ESI): 207.1 (M+H)+ .

[0341] Step 3: Preparation of Benzyl(R)-2-cyclopropyl-2-hydroxyacetate (40d) Compound 40c (12.1 g, 58.4 mmol) was purified by separating the chiral isomers using an SFC 150 preparative and a chiral column (chromatographic column: Daicel CHIRALCEL AD, 250 mm x 30 mm ID, 10 μm; mobile phase CO2 / MeOH [0.2% NH3 (7M Solution in MeOH)] = 90 / 10, flow rate: 120 g / min) to obtain compound 40d (5.8 g, yield: 48%). MS m / z (ESI): 207.1 (M+H) + .

[0342] Step 4: Preparation of (R)-10-cyclopropyl-1-(9H-fluoren-9-yl)-3,6-dioxy-2,9-dioxy-4,7-undecane-undecane-11-ate benzyl ester (40e) Compound 40d (5.8 g, 28 mmol) was dissolved in dichloromethane (20 mL), and (2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)acetamido)methyl acetate (5 g, 14 mmol) and 4-methylbenzenesulfonate pyridine (1.4 g, 5.6 mmol) were added. The reaction mixture was allowed to react at 40°C for 12 hours. After the reaction was complete, the mixture was concentrated under reduced pressure, and the resulting concentrate was purified by silica gel column chromatography system B to obtain the title compound 40e (1.2 g, yield: 30%). MS m / z (ESI): 537.2 (M+Na) + .

[0343] Step 5: Preparation of (R)-2-((2-aminoacetamide)methoxy)-2-cyclopropylbenzyl acetate (40f) Compound 40e (1.2 g, 2.33 mmol) was dissolved in N,N-dimethylformamide (10 mL), and diethylamine (0.34 g, 4.66 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the product was not purified and the next step was carried out directly. MS m / z (ESI): 293.1 (M+H) + .

[0344] Step 6: Preparation of (5S, 13R)-5-benzyl-13-cyclopropyl-1-(9H-fluoren-9-yl)-3,6,9-trioxo-2,12-dioxo-4,7,10-triazatetradecane-14-ate benzyl ester (40g) Compound 40f (0.6 g, 2.05 mmol) was dissolved in N,N-dimethylformamide (5 mL), and (((9H-fluoren-9-yl)methoxy)carbonyl)-L-phenylalanin lysine (1.09 g, 2.46 mmol) and N,N-diisopropylethylamine (0.53 g, 4.1 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the product was not purified and the next step was carried out directly. MS m / z (ESI): 684.3 (M+Na) + .

[0345] Step 7: Preparation of (R)-2-((2-((S)-2-amino-3-phenylpropionamide)acetamide)methoxy)-2-cyclopropylbenzyl acetate (40h) 40 g (1.2 g, 1.8 mmol) of the compound was dissolved in N,N-dimethylformamide (10 mL), and diethylamine (0.27 g, 3.6 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting concentrate was purified by silica gel column chromatography system B to obtain the title compound 40h (0.6 g, yield: 70%). MS m / z (ESI): 440.2 (M+H) + .

[0346] Step 8: Preparation of (11S, 19R)-11-benzyl-19-cyclopropyl-1-(9H-fluoro-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxy-4,7,10,13,16-pentazaeicosabenzyl carbonate (40i) Compound 40h (0.6 g, 1.36 mmol) was dissolved in N,N-dimethylformamide (10 mL), and (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycine (0.58 g, 1.63 mmol), N,N,N',N'-tetramethyl-O-(7-azabenzotriazole-1-yl)hexafluorophosphate urea (0.78 g, 2.04 mmol), and N,N-diisopropylethylamine (0.38 g, 2.72 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, water (10 mL) was added to the reaction mixture, and it was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting concentrate was purified by silica gel column chromatography system B to obtain the title compound 40i (0.8 g, yield: 76%). MS m / z (ESI): 798.2 (M+H) + .

[0347] Step 9: Manufacturing of (11S, 19R)-11-benzyl-19-cyclopropyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxy-4,7,10,13,16-pentazaeicosabenzyl carbonate (40j) Compound 40i (0.8 g, 1.03 mmol) was dissolved in a mixed solvent of ethanol and ethyl acetate (10 mL, V / V = 1:1), and Pd / C (0.2 g, 1.03 mmol) was added. The reaction mixture was stirred at room temperature under a hydrogen gas atmosphere for 2 hours. After the reaction was complete, the mixture was filtered through diatomaceous earth, the filter cake was washed with ethyl acetate, and the filtrates were combined and concentrated under reduced pressure to obtain the title compound 40j (0.55 g, yield: 71%). MS m / z (ESI): 708.1 (M+Na) + .

[0348] Step 10: Preparation of (2R, 10S)-16-amino-10-benzyl-2-cyclopropyl-6,9,12,15-tetraoxo-3-oxy-5,8,11,14-tetraazahexadecanoic acid (40k) Compound 40j (0.55 g, 0.8 mmol) was dissolved in N,N-dimethylformamide (10 mL), and diethylamine (117 mg, 1.6 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, tetrahydrofuran (20 mL) was added to the reaction mixture, the precipitated solid was filtered, and the filter cake was collected. The title compound 40k (0.2 g, yield: 54%) was obtained. MS m / z (ESI): 464.2 (M+H) + .

[0349] Step 11: Preparation of (2R, 10S)-10-benzyl-2-cyclopropyl-23-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)-6,9,12,15,18-pentaoxo-3-oxy-5,8,11,14,17-pentazatricarbon-22-ic acid (40L) Compound 40k (200 mg, 0.43 mmol) was dissolved in N,N-dimethylformamide (10 mL), and 2,5-dioxopyrrolidone-1-yl-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-ic acid ester 28f (152 mg, 0.39 mmol) and N,N-diisopropylethylamine (110 mg, 0.78 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, water (20 mL) was added to the reaction mixture, extracted with ethyl acetate (15 mL x 3), the organic phases were combined, washed with saturated brine, dried, and concentrated to obtain the crude product. The crude product was purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, chromatography column: Xtimate 10u C18 250 x 30 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 25%~95%, flow rate: 50 mL / min) to obtain 40 L (100 mg, yield: 35%) of the title compound. MS m / z (ESI): 766.2 (M+Na) + .

[0350] Step 12 Preparation of N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[yl]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxy-5,8,11,14-tetraazahexadecane-16-yl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-amide (40) 40 L (60 mg, 0.08 mmol) of compound was dissolved in N,N-dimethylformamide (6 mL), and compound 6m-2 (45 mg, 0.08 mmol), 4-(4,6-dimethoxytriazin-2-yl)-4-methylmorpholine hydrochloride (61 mg, 0.16 mmol), and triethylamine (26 mg, 0.16 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, water (20 mL) was added to the reaction mixture, extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with saturated brine, dried, concentrated, and the crude product was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5um C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 40%~100%, flow rate: 20 mL / min) to obtain the title compound 40 (10 mg, yield: 6%). MS m / z (ESI): 1175.3 (M+H) + . 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1H), 8.65 (d, 2H), 8.44 (s, 1H), 8.31 (s, 1H), 8.17 (d, 2H), 8.06 (s, 1H), 7.72 (d, 1H), 7.30 (s, 1H), 7.18 (dt, 5H), 6.52 (s, 1H), 5.60 (s, 1H), 5.44 (t, 3H), 5.30 (d, 1H), 4.82 (dd, 1H), 4.59(dd, 1H), 4.48 (dd, 1H), 4.07 (s, 3H), 3.76 - 3.66 (m, 5H), 3.60 - 3.53 (m, 2H), 3.39 (s, 3H), 3.23 (s, 3H), 3.00 (dd, 1H), 2.78 (d, 1H), 2.40 (d, 3H), 2.34 - 2.22 (m, 3H), 1.86 (qdd, 9H), 1.12 - 1.03 (m, 1H), 0.86 (t, 3H), 0.45 (d, 4H).

[0351] Example 41 .N-((2R,10S)-10-benzyl-2-cyclopropyl-1-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidine[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide [ka]

[0352] Step 1 N-((2R,10S)-10-benzyl-2-cyclopropyl-1-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indridine[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide Compound 035b (20 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (3 mL), and 40 L of compound (33 mg, 0.04 mmol), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (30 mg, 0.08 mmol), and N,N-dimethylethylenediamine (14 mg, 0.11 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction mixture was separated and purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: Gemini 5u C18 100 x 21.2 mm; mobile phase 1: water (containing 0.1% NH4); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 45%~75%, flow rate: 30 mL / min) to obtain the title compound 41 (18 mg, yield: 38%). MS m / z (ESI): 1177 (M+1) + . 1H NMR (400 MHz, CD3OD) δ 8.61 (s, 1H), 7.68-7.52 (m, 2H), 7.31-7.08 (m, 5H), 5.83-5.69 (m, 2H), 5.55 (dd, 2H), 5.40 (d, 1H), 4.68 (d, 2H), 4.60 (s, 2H), 4.44 - 4.35 (m, 2H), 4.28 (d, 1H), 4.12 (s, 3H), 3.80 - 3.71 (m, 4H), 3.48 (d, 2H), 3.17 - 3.06 (m, 3H), 2.97 - 2.85 (m, 2H), 2.73 (s, 1H), 2.58 (dd, 4H), 2.46 - 2.41 (m, 4H), 2.01 - 1.86 (m, 4H), 1.03 (dd, 3H), 0.94 (d, 1H), 0.49 - 0.40 (m, 2H), 0.34 (s, 2H).

[0353] Example 42 N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[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-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexa-5-inamide [ka]

[0354] Step 1 Preparation of 5-bromo-2-chloro-4-(methoxymethyl)pyrimidine (42b) 5-bromo-2-chloropyrimidine (10 g, 0.05 mol), silver nitrate (36 g, 0.2 mol), ammonium persulfate (57 g, 0.25 mol), and 2-methoxyacetic acid (5.4 g, 0.06 mol) were dissolved in acetonitrile (300 mL) and water (300 mL) and stirred at 60 °C for 2 hours. The reaction mixture was poured into water, extracted three times with ethyl acetate, washed once with saturated brine, concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography system B to obtain the title compound 42b (1.2 g, yield: 13%). MS m / z (ESI): 236.9 (M+1) + .

[0355] Step 2: Preparation of 6-(2-chloro-4-(methoxymethyl)pyrimidine-5-yl)hexyl-5-ic acid (42c) Compound 42b (1.2 g, 4.5 mmol) was dissolved in tetrahydrofuran (10 mL), and hexa-5-ic acid (0.76 g, 6.8 mmol), cuprous iodide (86 mg, 0.45 mmol), bistriphenylphosphine palladium dichloride (632 mg, 0.9 mmol), and triethylamine (1.4 g, 13.5 mmol) were added. The reaction mixture was stirred at 60 °C for 3 hours under a nitrogen gas atmosphere. After the reaction was complete, the reaction mixture was filtered, and the filtrate was stratified with ethyl acetate and water. The aqueous phase was extracted with ethyl acetate (20 mL x 3), the organic phases were combined, dried, and concentrated. The resulting crude product was purified by silica gel column chromatography system A to obtain the title compound 42c (600 mg, yield: 50%). MS m / z (ESI): 269.1 (M+1) + .

[0356] Step 3: 6-(4-(methoxymethyl)-2-(methylthio)pyrimidine-5-yl)hexyl-5-ic acid (42d) Compound 42c (600 mg, 2.2 mmol) was dissolved in dimethyl sulfoxide (6 mL), and sodium methyl mercaptan (154 mg, 2.2 mmol) and anhydrous magnesium sulfate (528 mg, 4.4 mmol) were added. The reaction mixture was stirred at 50°C for 1 hour. After the reaction was complete, the reaction mixture was poured into water, extracted with ethyl acetate (50 mL x 3), the organic phases were combined, dried, and concentrated. The resulting crude product 42d (600 mg) was then used directly in the next step without purification. MS m / z (ESI): 281.1 (M+1) + .

[0357] Step 4: Preparation of 6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-ic acid (42e) Compound 42d (600 mg) obtained in the previous step was dissolved in a mixed solvent of acetone and water (20 mL, V / V = 1:1), potassium peroxymonosulfate (7.6 g, 22 mmol) was added, and the mixture was stirred at room temperature for 1 hour. After complete reaction, the reaction mixture was poured into water, extracted three times with ethyl acetate, washed once with saturated brine, concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography system A to obtain the title compound 42e (200 mg, yield: 30%). MS m / z (ESI): 313.0 (M+1) + .

[0358] Step 5: Preparation of 2,5-dioxopyrrolidone-1-yl-6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidine-5-ylhexa-5-ic acid ester (42f) Compound 42e (200 mg, 0.64 mmol) and N-hydroxysuccinimide (110 mg, 0.96 mmol) were dissolved in dichloromethane (10 mL), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (246 mg, 1.28 mmol) was added. The reaction was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was directly concentrated to obtain the crude product, which was purified by silica gel column chromatography system B to obtain the title compound 42f (100 mg, yield: 38%). MS m / z (ESI): 410.1 (M+1) + .

[0359] Step 6: Preparation of (3R, 11S)-11-benzyl-24-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidine-5-yl)-3-methyl-7,10,13,16,19-pentaoxo-4-oxa-6,9,12,15,18-pentazatetracarbone-23-enoic acid (42g) Compound 42f (100 mg, 0.24 mmol) was dissolved in N,N-dimethylformamide (3 mL), and N,N-diisopropylethylamine (62 mg, 0.48 mmol) and compound 13j (108 mg, 0.24 mmol) were sequentially added to the solution. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was subjected to preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5u C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 30%~40%, flow rate: 20 mL / min) to obtain 42 g (30 mg, yield: 17%) of the title compound. MS m / z (ESI): 768.2 (M+Na) + .

[0360] Step 7 N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[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-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexa-5-inamide(42) 42 g (30 mg, 0.04 mmol) of the compound was dissolved in N,N-dimethylformamide (2 mL), compound 6m-2 (18 mg, 0.04 mmol) was added, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) and N,N-diisopropylethylamine (10 mg, 0.08 mmol) were added sequentially. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction mixture was separated and purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2*250 mm 10 μm; mobile phase 1: water (containing 0.1% NH4HCO3); mobile phase 2: acetonitrile; gradient for 18 minutes, gradient ratio: acetonitrile phase 38%~48%, flow rate: 30 mL / min) to obtain the title compound 42 (25 mg, yield: 53%). MS m / z (ESI): 1177.3 (M+1) + . 1H NMR (400 MHz, CD3OD) δ 8.88 (s, 1H), 7.55 (s, 1H), 7.48 (d, 1H), 7.18 (dd, 5H), 5.65 (dd, 1H), 5.57 (d, 1H), 5.36 (s, 2H), 5.32 (s, 1H), 4.75 (s, 2H), 4.73 (d, 1H), 4.64 (d, 1H), 4.50 (dd, 1H), 3.81 (t, 4H), 3.71 - 3.66 (m, 1H), 3.58 (d, 1H), 3.50 (s, 3H), 3.39 (s, 3H), 3.29 (d, 2H), 3.10 (dd, 1H), 2.89 (dd, 1H), 2.60 (ddd, 4H), 2.45 (s, 9H), 2.00 - 1.87 (m, 6H), 1.27 (d, 3H), 1.00 (t, 3H).

[0361] Example 43 N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[]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-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-amide [ka]

[0362] Step 1: Preparation of (2R, 10S)-10-benzyl-2-cyclopropyl-23-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidine-5-yl)-6,9,12,15,18-pentaoxo-3-oxa-5,8,11,14,17-pentazatricarbon-22-ic acid (43a) Compound 42f (25 mg, 0.06 mmol) was dissolved in N,N-dimethylformamide (3 mL), and N,N-diisopropylethylamine (62 mg, 0.48 mmol) and compound 40k (28 mg, 0.06 mmol) were sequentially added to the solution, and the reaction was stirred at room temperature for 3 hours. The reaction mixture was subjected to preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5u C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 20 minutes, gradient ratio: acetonitrile phase 35%~40%, flow rate: 20 mL / min) to obtain the title compound 43a (7 mg, yield: 15%). MS m / z (ESI): 780.2 (M+Na) + .

[0363] Step 2 N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[yl]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-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-amide Compound 43a (7 mg, 0.01 mmol) was dissolved in N,N-dimethylformamide (2 mL), and compound 23b (5 mg, 0.01 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (6 mg, 0.02 mmol), and N,N-diisopropylethylamine (4 mg, 0.03 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction mixture was separated and purified by high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2*250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 18 minutes, gradient ratio: acetonitrile phase 35%~65%, flow rate: 30 mL / min) to obtain the title compound 43 (5 mg, yield: 45%). MS m / z (ESI): 1215.3 (M+1) + . 1 H NMR (400 MHz, CD3OD) δ 8.85 (s, 1H), 8.52 (s, 1H), 7.60 (s, 1H), 7.18 (dt, 4H), 5.67 (d, 2H), 5.55 (dd, 4H), 5.42 - 5.31 (m, 4H), 4.64 (d, 2H), 3.85 (d, 2H), 3.82 (s, 2H), 3.77 (d, 2H), 3.57 (s, 2H), 3.46 (s, 3H), 3.36 (s, 4H), 2.60 (t, 2H), 2.43 (d, 2H), 2.19 (d, 2H), 1.96 - 1.89 (m, 5H), 1.16 (d, 4H), 0.98 (t, 4H), 0.88 (d, 1H), 0.81 (d, 1H), 0.72 (s, 1H), 0.62 (s, 1H), 0.49 (d, 2H), 0.37 - 0.32 (m, 1H).

[0364] Example 44 N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[yl]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-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-inamide [ka]

[0365] Step 1 N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[yl]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-(4-methoxy-2-(methylsulfonyl)pyrimidine-5-yl)hexyl-5-inamide(44) Compound 40L (30 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (2 mL), and compound 23b (19.2 mg, 0.040 mmol), Castro condenser (21.4 mg, 0.048 mmol), and N,N-diisopropylethylamine (10.4 mg, 0.081 mmol) were added. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the reaction mixture was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2*250 mm 10 μm; mobile phase 1: water (containing 0.1% NH4HCO3); mobile phase 2: acetonitrile; gradient for 18 minutes, gradient ratio: acetonitrile phase 46%~100%, flow rate: 30 mL / min) to obtain the title compound 44 (4.2 mg, yield: 9%). MS m / z (ESI): 1201.4 (M+1) + . 1 H NMR (400 MHz, CD3OD) δ 8.56 (s, 1H), 7.59 (s, 1H), 7.57 (d, 1H), 7.24 - 7.16 (m, 4H), 7.15 - 7.10 (m, 1H), 5.71 - 5.66 (m, 1H), 5.60-5.50 (m, 2H), 5.43-5.31 (m, 2H), 4.96 (d, 2H), 4.68-4.57 (m, 1H), 4.51 (dd, 1H), 4.09 (s, 3H), 3.93 - 3.71 (m, 6H), 3.58 (d, 3H), 3.15-3.10 (m, 1H), 2.97-2.92 (m, 1H), 2.62-2.40 (m, 5H), 2.26 - 2.14 (m, 2H), 2.02-1.74 (m, 6H), 1.37-1.28 (m, 2H), 1.21-1.09 (m, 3H), 0.98 (t, 3H), 0.86- 0.29 (m, 6H).

[0366] Example 45 N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[yl]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxy-5,8,11,14-tetraazahexadecane-16-yl)-6-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexa-5-inamide [ka]

[0367] Step 1: Preparation of ((2R, 10S)-10-benzyl-23-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)-2-cyclopropyl-6,9,12,15,18-pentaoxo-3-oxa-5,8,11,14,17-pentazatricarbon-22-ic acid (45a) Compound 42f (100 mg, 0.25 mmol) was dissolved in N,N-dimethylformamide (2 mL), and compound 40k (130 mg, 0.28 mmol) and N,N-diisopropylethylamine (66 mg, 0.51 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: Gemini 5u C18 100 x 21.2 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 42%~100%, flow rate: 25 mL / min) to obtain the title compound 45a (100 mg, yield: 53%). MS m / z (ESI): 760.0 (M+Na) + .

[0368] Step 2: Preparation of N-((2R, 10S)-10-benzyl-2-cyclopropyl-1-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[yl]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxy-5,8,11,14-tetraazahexadecane-16-yl)-6-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexa-5-inamide (45) Compound 45a (115 mg, 0.16 mmol) was dissolved in N,N-dimethylformamide (2 mL), and compound 6m-2 (70 mg, 0.16 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (89 mg, 0.23 mmol), and N,N-dimethylformamide (40 mg, 0.31 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5 um C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 50%~65%, flow rate: 20 mL / min) to obtain the title compound 45 (60 mg, yield: 33%). MS m / z (ESI): 1169.2 (M+1) + . 1H NMR (400 MHz, DMSO-d6) δ 8.96 (s, 1H), 8.70 (s, 1H), 8.67 (d, 1H), 8.57 (d, 1H), 8.34-8.22 (m, 1H), 8.21-8.14 (m, 1H), 8.11 (d, 1H), 8.07-8.00 (m, 1H), 7.72 (d, 1H), 7.30 (s, 1H), 7.25-7.17 (m, 4H), 7.17-7.10 (m,1H), 6.52 (s, 1H), 5.60 (s, 1H), 5.44 (t, 2H), 5.30 (d, 1H), 4.86-4.78 (m, 1H), 4.63-4.55 (m, 1H), 4.53-4.45 (m, 1H), 3.71- 3.66 (m, 4H), 3.60 (d, 1H), 3.55 (d, 2H), 3.44 (s, 2H), 3.24-3.20 (m, 2H), 3.04 -2.97 (m, 2H), 2.67 (s, 1H), 2.40 (s, 3H), 2.11-2.04 (m, 2H), 2.03-1.92 (m, 3H), 1.88-1.76 (m, 4H), 1.75 -1.62 (m, 2H), 1.46 (s, 1H), 1.13 -1.02 (m, 1H), 0.86 (t, 4H), 0.50 - 0.39 (m, 4H).

[0369] Example 46 N-((2R,10S)-10-benzyl-2-cyclopropyl-1-((1S,10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[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-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexa-5-inamide(46) [ka]

[0370] Step 1 N-((2R,10S)-10-benzyl-2-cyclopropyl-1-((1S,10S)-5-cyclopropyl-10-ethyl-6-fluoro-10-hydroxy-11,14-dioxo-1,2,3,4,10,14,16-octahydro-13H-cyclohepta[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-(5-cyano-6-(methylsulfonyl)pyridine-3-yl)hexa-5-inamide Compound 45a (16 mg, 0.02 mmol) was dissolved in N,N-dimethylformamide (2 mL) and added to compound 23b (10 mg, 0.02 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11 mg, 0.03 mmol), and N,N-diisopropylethylamine (5 mg, 0.04 mmol). The reaction was stirred at room temperature for 30 minutes. The reaction mixture was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: WELCH Xtimate C18 21.2*250 mm 10 μm; mobile phase 1: water (containing 0.1% NH4HCO3); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 45%~55%, flow rate: 25 mL / min) to obtain the title compound (9 mg, yield: 36%). MS m / z (ESI): 1195.3 (M+1) + . 1H NMR (400 MHz, CD3OD) δ 8.76 (dd, 1H), 8.35 (dd, 1H), 7.62 - 7.56 (m, 1H), 7.51 (d, 1H), 7.19 (s, 5H), 5.68 (s, 2H), 5.40 (t, 2H), 5.16 - 5.07 (m, 2H), 4.94 (d, 2H), 4.60 (dd, 2H), 4.46 (d, 2H), 3.86 (d, 2H), 3.75 (d, 2H), 3.68 (d, 1H), 3.59 -3.56 (m, 2H), 3.36 (d, 3H), 3.35 (s, 1H), 2.94 (dd, 2H), 2.57 - 2.53 (m, 1H), 2.44 (d, 2H), 2.34 (d, 2H), 2.20 (s, 2H), 1.91 (s, 2H), 1.80 (s, 2H), 1.04 (d, 2H), 0.99 (t, 1H), 0.81 (s, 2H), 0.72 (s, 2H), 0.50 (s, 2H), 0.36 (s, 2H).

[0371] Example 47 N-((2R,10S)-10-benzyl-2-cyclopropyl-1-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indolidine[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide [ka]

[0372] Step 1 N-((2R,10S)-10-benzyl-2-cyclopropyl-1-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-oxepino[4,3,2-de]pyrano[3',4':6,7]indridine[1,2-b]quinoline-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide Compound 035b (20 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (3 mL), and compound 43a (34 mg, 0.04 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (30 mg, 0.08 mmol), and N,N-diisopropylethylamine (14 mg, 0.11 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction mixture was separated and purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: Gemini 5u C18 100 x 21.2 mm; mobile phase 1: water (containing 0.1% NH4); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 30%~60%, flow rate: 30 mL / min) to obtain the title compound (19 mg, yield: 40%). MS m / z (ESI): 1191.2 (M+1) + . 1H NMR (400 MHz, CD3OD) δ 8.85 (d, 1H), 7.56 (dd, 2H), 7.27-7.10 (m, 5H), 5.78 -5.67 (m, 2H), 5.52 (dd, 2H), 5.37 (d, 1H), 4.74 - 4.56 (m, 7H), 4.42-4.23 (m, 3H), 3.75 (d, 2H), 3.72-3.67 (m, 2H), 3.46 (d, 3H), 3.37 (s, 3H), 3.07 (dd, 1H), 2.94-2.86 (m, 1H), 2.77-2.69 (m, 1H), 2.66-2.55 (m, 3H), 2.45-2.39 (m, 3H), 1.97-1.89 (m, 3H), 1.31 (d, 3H), 1.00 (dd, 3H), 0.94-0.88 (m, 1H), 0.43 (dd, 2H), 0.37-0.27 (m, 2H).

[0373] Example 48 (R)-3-((((S)-7-benzyl-17-(4-(2-(methylsulfonyl)-4-(trifluoromethyl)pyrimidine-5-yl)piperidine-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentazaheptadecyl)oxy)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)butanamide [ka]

[0374] Step 1 4-(2-(methylthio)-4-(trifluoromethyl)pyrimidine-5-yl)piperidine-1-carboxylate tert-butyl(48a) Nickel dimethoxyethane chloride (350 mg, 1.6 mmol) and 2-carboxyimidoamide hydrochloride (250 mg, 1.6 mmol) were added to a reaction flask, and under a nitrogen atmosphere, N,N-dimethylacetamide (10 ml) was added, and the mixture was stirred at room temperature for 30 minutes. In a separate reaction flask, 4-iodopiperidine-1-carboxylate tert-butyl 29a (2.0 g, 6.4 mmol), 5-bromo-2-(methylthio)-4-(trifluoromethyl)pyrimidine (1.75 g, 6.4 mmol), manganese (90 mg, 1.6 mmol), and tetrabutylammonium iodide (2.36 g, 6.4 mmol) were added, followed by N,N-dimethylacetamide (10 ml), and the mixture was substituted with nitrogen gas three times. After substitution was complete, the nickel ligand compound was transferred to the system using a syringe, and the reaction was carried out at 60°C for 3 hours. After the reaction was complete, ethyl acetate (200 mL) and water (400 mL) were added to the system for extraction. The organic phase was dried, concentrated, and purified using silica gel column chromatography separation system B to obtain the title compound 48a (1.3 g, yield: 48%). MS m / z (ESI): 322.0 (M-55) + .

[0375] Step 2: 2-(methylthio)-5-(piperidine-4-yl)-4-(trifluoromethyl)pyrimidine (48b) Compound 48a (1.3 g, 3.4 mmol) was dissolved in 10 ml of 1,4-dioxane solution in 4N hydrochloric acid. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the solvent was directly evaporated by rotation to obtain crude product 48b (800 mg, yield: 85%). MS m / z (ESI): 278.0 (M+1) + .

[0376] Step 3: 3-(4-(2-(methylthio)-4-(trifluoromethyl)pyrimidine-5-yl)piperidine-1-yl) tert-butyl (48c) propionate Compound 48b (800 mg, 2.9 mmol) was dissolved in acetonitrile (10 mL), and tert-butyl 3-bromopropionate (910 mg, 4.35 mmol) and potassium carbonate (1.2 g, 8.7 mmol) were added. The reaction was stirred at 60°C for 16 hours. After the reaction was complete, the solvent was evaporated by rotation to obtain the crude product, which was purified by column chromatography separation system B to obtain the title compound 48c (600 mg, yield: 51%). MS m / z (ESI): 406.1 (M+1) + .

[0377] Step 4 3-(4-(2-(methylsulfonyl)-4-(trifluoromethyl)pyrimidine-5-yl)piperidine-1-yl)propionic acid (48d) Compound 48c (600 mg, 1.48 mmol) was dissolved in trifluoroacetic acid (10 mL), and metachloroperbenzoic acid (2.55 g, 14.8 mmol) was added. The reaction was carried out at 40°C for 4 hours. After the reaction was complete, the mixture was concentrated, and dichloromethane (80 ml) and water (80 ml) were added for extraction. The aqueous phase was freeze-dried, and the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5u C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 15 minutes, gradient ratio: acetonitrile phase 10%~20%, flow rate: 20 mL / min) to obtain the title compound 48d (230 mg, yield: 41%). MS m / z (ESI): 382.0 (M+1) + .

[0378] Step 5: 2,5-Dioxopyrrolidone-1-yl-3-(4-(2-(methylsulfonyl)-4-(trifluoromethyl)pyrimidine-5-yl)piperidine-1-yl)propionate (48e) Compound 48d (50 mg, 0.13 mmol) was dissolved in dimethyl sulfoxide (1 mL), and N-hydroxysuccinimide (17 mg, 0.14 mmol) and EDCI (38 mg, 0.20 mmol) were added. The reaction was carried out at room temperature for 16 hours. After the reaction was complete, the reaction solution was used directly in the next step. MS m / z (ESI): 479.1 (M+1) + .

[0379] Step 6 (3R,11S)-11-benzyl-3-methyl-21-(4-(2-(methylsulfonyl)-4-(trifluoromethyl)pyrimidine-5-yl)piperidine-1-yl)-7,10,13,16,19-pentaoxo-4-oxa-6,9,12,15,18-pentazaeicosanoic acid (48f) Compound 13j (58 mg, 0.13 mmol) was added to the reaction mixture from the previous step, and N,N-diisopropylethylamine (17 mg, 0.26 mmol) was added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction mixture was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatography column: Xbridge 5u C18 150 x 19 mm 5um; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient for 9.5 minutes, gradient ratio: acetonitrile phase 20%~30%, flow rate: 20 mL / min) to obtain the title compound 48f (20 mg, yield: 19%). MS m / z (ESI): 815.2 (M+1) + .

[0380] Step 7 (R)-3-((((S)-7-benzyl-17-(4-(2-(methylsulfonyl)-4-(trifluoromethyl)pyrimidine-5-yl)piperidine-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentazaheptadecyl)oxy)-N-((1S, 10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)butanamide Compound 48f (20 mg, 0.025 mmol) was dissolved in N,N-dimethylformamide (1 mL), and compound 6m-2 (11 mg, 0.025 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (14 mg, 0.037 mmol), and N,N-diisopropylethylamine (6 mg, 0.049 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction was complete, the reaction solution was purified by preparative high-performance liquid chromatography (Waters MS-triggered Pre LC with QDA detector, chromatography column: Gemini 5u C18 100 x 21.2 mm 5um; mobile phase 1: water (containing 0.1% NH4HCO3); mobile phase 2: acetonitrile; gradient for 9.5 minutes, gradient ratio: acetonitrile phase 35%~65%, flow rate: 30 mL / min) to obtain the title compound 48 (3.9 mg, yield: 12%). MS m / z (ESI): 1246.5 (M+1) + . 1H NMR (400 MHz, CD3OD) δ 9.40 (s, 1H), 7.54 (s, 1H), 7.50 (d, 1H), 7.15-7.07 (m, 5H), 5.64 (dd, 1H), 5.55 (d, 1H), 5.35 (s, 2H), 5.33-5.27 (m, 1H), 4.70 (d, 1H), 4.59 (d, 1H), 4.52 (dd, 1H), 4.2-4.14 (m, 1H), 3.85 (s, 3H), 3.84-3.68 (m, 3H), 3.56 (d, 1H), 3.38 (s, 3H), 3.15-3.12 (m, 2H), 3.09 (d, 1H), 3.05 (d, 1H), 2.85 (dd, 1H), 2.74-2.68 (m, 2H), 2.55 (d, 1H), 2.43 (d, 4H), 2.21-2.10 (m, 4H), 2.06-1.98 (m, 3H), 1.91-1.86 (m, 2H), 1.84-1.78 (m, 2H), 1.60 (s, 1H), 1.34-1.32 (m, 2H), 1.24 (d, 3H), 0.96 (t, 3H), 0.89 (d, 1H).

[0381] Manufacturing of antibody complexes

[0382] Example A-1 Manufacturing of ADC-1

change

[0383] Trastuzumab was replaced with 100 mM Pro-Ac, 20 mM Arg, and pH 5.0 using Amicon. Trastuzumab was removed, and 1 M trometamol and pH 7.5 were added to adjust the pH to 7.3. Trastuzumab (67 mg) was placed in a 50 mL centrifuge tube, and 2801 μL of 100 mM acetylproline, 20 mM arginine, pH 7.3, 96 μL of 0.2 M EDTA, and 893 μL of 5 mM TCEP were added. The mixture was homogeneously mixed in a 37°C shaker and allowed to react for 1 hour. After the reaction was complete, the reduced trastuzumab product was purified using a Zeba desalting column.

[0384] Trastuzumab reduced product (47.40 mg) was placed in a 50 mL centrifuge tube, and under ice bath, 1103.58 μL of 100 mM Pro-Ac, 20 mM Arg, pH 7.3, 94.80 μL of 0.2 M EDTA, 790 μL of DMA, and 632 μL of 10 mM compound 21 were added. The mixture was homogeneously mixed in a 22°C shaker and reacted for 3 hours. The reaction mixture was purified using a Zeba desalting column, the sample was concentrated using Amicon, and the solution was replaced with a buffer system of 100 mM proline, 20 mM arginine, pH 5.0 to obtain ADC-1 (33 mg). The mean DAR value of ADC was calculated by LC-MS and β = 8.14.

[0385] Referring to the method of Example A-1, compound 21 was replaced with another compound represented by formula V of this application to produce the following antibody-drug conjugate, the structure and DAR value (β) of which are shown in the table below. [Table 1] [Table 2] [Table 3]

[0386] The cytotoxic drug-connector compound represented by formula V of the present invention can be smoothly conjugated with an antibody to obtain an antibody-drug conjugate.

[0387] Biological evaluation

[0388] The present invention will be further described below with reference to test examples, but these test examples are not intended to limit the scope of the present invention.

[0389] Experiment Example 1: Experiment on inhibition of SK-BR-3 cell proliferation by a compound.

[0390] 1: Experimental materials [Table 4]

[0391] 2: Laboratory equipment [Table 5]

[0392] 3: Test Method

[0393] (1) Cell Plating: First, tumor cells SK-BR-3 were cultured in the appropriate medium, digested with trypsin, resuspended and counted after centrifugation, and then plated in 384-well plates after adjusting the cell concentration to the appropriate level.

[0394] (2) Co-incubation of compounds and tumor cells: After the cells adhered, 100 nL of diluted biologically active substance (test compound) was added to the cell culture plate by ultrasound, and the final concentration of DMSO in the wells of the cell culture plate was 0.33%, and the cells were incubated for 72 hours in an incubator at 37°C and 5% CO2.

[0395] (3) After incubation was complete, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the mixture was placed in a high-speed shaker and shaken for 2 minutes. Then it was centrifuged at 1000 rpm for 1 minute, left in the dark at room temperature for 30 minutes, and the chemiluminescence signal value was read using an Envision instrument.

[0396] (4) Detection of cell activity: IC with GraphPad Prism 8 software 50 The IC of the compound is calculated and the following nonlinear fitting equation is used. 50 Obtain (see Table 1).

[0397] 1.Y=Bottom + (Top-Bottom) / (1+10^((LogIC 50 -X)* HillSlope)) 2. Y: Inhibition rate, X: Compound concentration log value, 3. Inhibition rate (%) = 100 - (Reading in compound well - Reading in low-reading control well) / (Reading in high-reading control well - Reading in low-reading control well) * 100; 4. High-reading control well: 100 nL of DMSO was added to the cells. 5. Low-reading control wells: Cell-free wells. [Table 6] [Table 7]

[0398] As the results show, the compounds of the present invention have a strong inhibitory effect on the proliferation of SK-BR-3 cells.

[0399] Experiment Example 2: Experiment on inhibition of MDA-MB-468 cell proliferation by test compounds.

[0400] 1: Experimental materials [Table 8]

[0401] 2: Laboratory equipment [Table 9]

[0402] 3: Test Method (1) Cell Plating: First, tumor cells MDA-MB-468 were cultured in the appropriate medium, digested with trypsin, resuspended and counted after centrifugation, and then plated in 384-well plates after adjusting the cell concentration to the appropriate level.

[0403] (2) Co-incubation of compounds and tumor cells: After the cells adhered, 100 nL of diluted biologically active substance (test compound) was added to the cell culture plate by ultrasound, and the final concentration of DMSO in the wells of the cell culture plate was 0.33%, and the cells were incubated for 72 hours in an incubator at 37°C and 0% CO2.

[0404] (3) After incubation was complete, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the mixture was placed in a high-speed shaker and shaken for 2 minutes. Then it was centrifuged at 1000 rpm for 1 minute, left in the dark at room temperature for 30 minutes, and the chemiluminescence signal value was read using an Envision instrument.

[0405] (4) Detection of cell activity: IC with GraphPad Prism 8 software 50 The IC of the compound is calculated and the following nonlinear fitting equation is used. 50 Obtain (see Table 2).

[0406] 6.Y=Bottom + (Top-Bottom) / (1+10^((LogIC 50 -X)* HillSlope)) 7. Y: Inhibition rate, X: Compound concentration log value, 8. Inhibition rate (%) = 100 - (Reading in compound well - Reading in low-reading control well) / (Reading in high-reading control well - Reading in low-reading control well) * 100; 9. High-reading control well: 100 nL of DMSO was added to the cells. 10. Low-reading control wells: Cell-free wells. [Table 10] [Table 11]

[0407] As the results show, the test compound has a strong inhibitory effect on the proliferation of MDA-MB-468 cells.

[0408] Experiment Example 3: Experiment on inhibition of NCI-N87 cell proliferation by test compounds.

[0409] 1: Experimental materials [Table 12]

[0410] 2: Laboratory equipment [Table 13]

[0411] 3: Test Method (1) Cell Plating: First, tumor cells NCI-N87 were cultured in the appropriate medium, digested with trypsin, resuspended and counted after centrifugation, and then plated in 384-well plates after adjusting the cell concentration to the appropriate level.

[0412] (2) Co-incubation of compounds and tumor cells: After the cells adhered, 100 nL of diluted biologically active substance (test compound) was added to the cell culture plate by ultrasound, and the final concentration of DMSO in the wells of the cell culture plate was 0.33%, and the cells were incubated for 72 hours in an incubator at 37°C and 0% CO2.

[0413] (3) After incubation was complete, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the mixture was placed in a high-speed shaker and shaken for 2 minutes. Then it was centrifuged at 1000 rpm for 1 minute, left in the dark at room temperature for 30 minutes, and the chemiluminescence signal value was read using an Envision instrument.

[0414] (4) Detection of cell activity: IC with GraphPad Prism 8 software 50 The IC of the compound is calculated and the following nonlinear fitting equation is used. 50 Obtain (see Table 3).

[0415] 11.Y=Bottom + (Top-Bottom) / (1+10^((LogIC 50 -X)* HillSlope)) 12. Y: Inhibition rate, X: Compound concentration log value, 13. Inhibition rate (%) = 100 - (Reading in compound well - Reading in low-reading control well) / (Reading in high-reading control well - Reading in low-reading control well) * 100; 14. High-reading control well: 100 nL of DMSO was added to the cells. 15. Low-reading control wells: Cell-free wells. [Table 14] [Table 15]

[0416] As the results show, the test compound has a strong inhibitory effect on the proliferation of NCI-N87 cells.

[0417] Test Example 4: Dynamic solubility test in PBS 7.4 buffer.

[0418] 4.1 Experimental Steps

[0419] 1) Preparation of stock solution

[0420] Using DMSO, 10 mM stock solutions were prepared for the target substances 6-2, 11-2, 17-2, Dxd, and the control drugs progesterone and diclofenac, respectively.

[0421] 2) Steps for measuring dynamic solubility

[0422] 15 μL of 10 mM stock solution was taken and added to the corresponding positions in the 96-well plate in the specified order. 485 μL of PBS 7.4 buffer was added to the corresponding vial in the sample plate. The experiment was performed in duplicate. One stirring rod was added to each vial, and the vials were capped. The sample tray was then placed in a thermomixer and shaken for 2 hours at a rotation speed of 1100 rpm and 25°C. After 2 hours, the caps were removed, the stirring rods were aspirated with a large magnet, and the samples were then transferred from the sample plate to a filter plate. Negative pressure was generated with a vacuum pump, and the samples were filtered. 5 μL of filtrate and 5 μL of blank DMSO were transferred to a new sample plate, and then 490 μL of internal standard water (acetonitrile:water = 1:1) containing the internal standard was added. Depending on the peak shape, the sample diluent can be diluted with a specific proportion of internal standard water to obtain a better peak.

[0423] 3) Preparation of 3 μM standard solution

[0424] 6 μL of DMSO stock solution was transferred from a 10 mM DMSO stock solution plate to an empty plate, and 194 μL of DMSO was added to prepare a 300 μM standard solution. 5 μL of the 300 μM standard solution plate was transferred to another empty plate, and 490 μL of internal standard water (acetonitrile:water = 1:1), including 5 μL of blank buffer and the internal standard, was added until the final concentration of the standard solution was 3 μM.

[0425] 4) Sample analysis step

[0426] The sample plates were placed in the sample tray of an autosampler, and the samples were analyzed and evaluated using liquid chromatography.

[0427] 4.0 Data Analysis

[0428] All calculations were performed in Microsoft Excel.

[0429] The analysis and quantification of the sample filtrate are performed by qualitative and quantitative analysis of standard peaks of known concentrations using liquid chromatography. The formulas for calculating the solubility values ​​of the control drug and the substance being measured are as follows.

number

[0430] [Sample] is the compound concentration, DF Sample [STD] refers to the sample dilution factor, and AREA is the concentration of the compound standard. Std This represents the peak area of ​​the compound standard, AREA Sample This represents the peak area of ​​the compound sample.

[0431] 4.2 Experimental Results

[0432] The experimental results are shown in Table 4. [Table 16]

[0433] As the results show, the solubility of representative compounds 6-2 and 11-2 of this application in PBS 7.4 buffer is clearly superior to that of Dxd.

[0434] Study Example 5: Pharmacokinetic Study in SD Rats

[0435] In male SD rats that received a single intravenous injection of compounds 6-2, 11-2, 17-2, and Dxd (dose: 2 mpk), the mean pharmacokinetic parameters in plasma were as shown in Table 5 (blood sampling times: 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 8h, and 24h after IV administration, a total of 8 blood sampling times). [Table 17]

[0436] As the results show, the representative compound of this application is rapidly eliminated from the rat body, has good safety, and possesses good pharmacokinetic properties.

[0437] Experiment Example 6: Inhibition of human liver microsomal CYP1A2, CYP2D6, and CYP3A4 by compound

[0438] 6.1 Subtype-positive control inhibitors [Table 18]

[0439] 6.2 Preparation of substrate stock solution [Table 19]

[0440] 6.3 Experimental Process

[0441] Incubation was performed in a 96-deep-well plate. 169 μL of the "main solution" and 1 μL of multiple concentrations of the test compound or positive control compound (DMSO) were added to each well of the incubation plate. The incubation plate was pre-incubated in a water bath pan at 37°C for 5 minutes. Then, 10 μL of diluted substrate solution was added to the incubation plate and incubated for 15 seconds while mixing with a vortex mixer. A further 20 μL of 10 mM NADPH solution was added, and the reaction was initiated when the final concentration reached 1 mM. At the predetermined time, 300 μL of stop solution (cold acetonitrile + 3% formic acid, 200 nM alprazolam, 200 nM labetalol hydrochloride, 200 nM tolbutamide) was added to quench the reaction. The mixture was centrifuged at 3220 g for 40 minutes. 150 μL of the supernatant was transferred to a new plate. The supernatant may be diluted with 150 μL of pure water. The mixture was homogenized, and the content of substrate metabolites was measured by LC / MS / MS.

[0442] 6.4 Data Analysis

[0443] Automated peak integration region checks were performed on all samples. Analytical peak area and internal standard peak area were exported to an Excel sheet. Inhibition of each P450 enzyme in human liver microsomes was measured by the percentage decrease in marker metabolite formation activity compared to the non-inhibited control (=100% activity). IC was calculated using the logarithm of residual activity (%) and inhibitor concentration. 50 The value was calculated.

[0444] The percentage of remaining activity is calculated as follows: Area ratio = Peak area of ​​analyte / Peak area of ​​internal standard Residual activity (%) = area ratio 測定対象薬物 / area ratio ブランク対照 *100% Using Excel XLfit 5.5.1.3 to create an IC 50 The value was calculated.

[0445] The experimental results are shown in Table 6. [Table 20]

[0446] As the results show, the representative compounds of this application exhibit low inhibitory activity on CYP enzymes and have good safety.

[0447] Test Example 7: Stability Test of Compounds in Human Plasma

[0448] 7.1 Experimental Process

[0449] (1) 199 microliters of human plasma were added to each cell culture plate, the culture plates were preheated to 37°C, and kept warm for 15 minutes.

[0450] (2) After pre-incubation, 1 microliter of the 1 mmol / l test compound and 1 microliter of the 1 mmol / l control compound were added to 199 microliters of plasma to achieve final concentrations of 5 μmol / l for the test compound and 5 μmol / l for the control compound. The final concentration of the organic solvent was 0.5%. The experiment was repeated twice.

[0451] (3) The reaction samples were incubated at 37°C.

[0452] (4) The reaction was stopped at 0, 1, 2, 6, and 24 hours by adding 600 μL of cold methanol containing an internal standard. All samples were vortexed for 10 minutes, followed by centrifugation at 3220 g for 30 minutes to precipitate the protein. 100 μL of supernatant was transferred to a new plate. The supernatant was diluted with ultrapure water according to the signal response and peak waveform of liquid chromatography-mass spectrometry (LC-MS).

[0453] 7.2 Data Analysis

[0454] All calculations were performed using Microsoft Excel. Peak area ratios were determined from the extracted ion chromatograms. The percentage of the remaining compound at each time point was calculated using the following formula: Remaining percentage tmin (%) = Peak Area Ratio tmin / Peak area t0 × 100

[0455] The experimental results are shown in 7. [Table 21]

[0456] As the results show, representative compounds 6-2, 11-2, and 17-2 of this application exhibit good stability in human plasma.

[0457] Test Example 8: Detection of ADC Biological Activity

[0458] 1. Test Objectives

[0459] The objective of this experiment was to investigate the inhibitory activity of ADC compounds on the in vitro proliferation of HER2-expressing Calu-3 cells, SK-BR-3 cells, and HER2-negative MDA-MB-468 cells. Cells were treated in vitro with different concentrations of the compound and cultured for 5 days. Cell proliferation was then detected using the CTG CellTiter-Glo® Luminescent Cell Viability Assay. 50 The in vitro activity of the compound was evaluated based on the values.

[0460] 2. Test method:

[0461] (1) On day 1, tumor cells were plated into a 96-well plate, with 5000 cells / 100 μL of medium inoculated into each well, and 100 μL of DPBS inoculated into each empty well at the edge. The plates were incubated overnight in a 37°C incubator.

[0462] (2) On the second day, first aspirate the old culture medium at a rate of 50 μL / well, and add ADC with different concentration gradients. The ADC has an initial concentration of 200 nM, which is diluted fivefold to 9 different concentrations. The drug administration volume is 50 μL / well.

[0463] (3) On day 6, the CellTiter-Glo Buffer and CellTiter-Glo Substrate reagents were thawed at 4°C. Before use, 10 ml of Buffer was drawn out, added to the Substrate, mixed uniformly, and allowed to equilibrate to room temperature.

[0464] (4) On day 7, the 96-well plate was equilibrated at room temperature for 30 minutes, and 100 μL of Cell-Titer-Glo was added to each well. After shaking for 5 minutes in the dark at room temperature and allowing to stand for 10 minutes, 100 μL of the liquid in each well was transferred to a white plate, and chemiluminescence was detected with a microblade reader.

[0465] 3. Data Analysis

[0466] Data was processed and analyzed using Microsoft Excel and Graphpad Prism 5 to test the inhibitory activity of ADC compounds on the in vitro proliferation of Calu-3 cells, SK-BR-3 cells, and MDA-MB-468 cells, as shown in Table 8 below. [Table 22]

[0467] Conclusion: The antibody-drug conjugate of the present invention, which targets HER2, exhibits clear growth inhibitory activity against HER2-positive cells SK-BR-3 and Calu-3, while also exhibiting weak growth inhibitory activity against HER2-negative cells MDA-MB-468, demonstrating good selectivity.

[0468] Test Example 9: Her2-ADC Plasma Stability Experiment

[0469] The mice used in this experiment are CD-1 mice, the rats are SD rats, and the monkeys are cynomolgus macaques.

[0470] (1) Free toxin release test and results

[0471] DS8201 sample, ADC-5, and ADC-10 were added to the above-mentioned sterile mouse plasma, sterile rat plasma, sterile human plasma, and sterile monkey plasma at a final concentration of 200 μg / mL, respectively. ADC-14 and ADC-15 were also added to the above-mentioned sterile mouse plasma and sterile human plasma at a final concentration of 200 μg / mL. These samples were incubated in a cell incubator at 37°C, with the incubation day designated as day 0. Subsequently, samples were taken out on days 1, 4, 7, 14, and 21, respectively, and the free toxin content was detected.

[0472] According to the results of free toxin release, ADC-5 and ADC-10 were quite stable in mouse, rat, human, and monkey plasma, with a maximum free toxin release rate of 0.3% or less, which is significantly better than the reference DS8201 (see Figures 1-3).

[0473] According to the free toxin release results, ADC-5 and ADC-10 showed superior free toxin release rates compared to ADC-14 and ADC-15 in both mouse and human plasma (see Figures 4-5). It was shown that the introduction of appropriate substituents (e.g., methoxy groups) to the pyrimidine ring can reduce safety issues of ADCs due to toxin elimination by improving the stability of ADCs in plasma.

[0474] (2) Test and results of ADC DAR values

[0475] DS8201 samples, ADC-5, ADC-10, and ADC-13 were added to the above-mentioned sterile human plasma at a final concentration of 200 ug / mL and incubated in a cell incubator at 37°C. The incubation day was designated as day 0, and then samples were taken out on days 1, 4, 7, 14, and 21, respectively, and changes in DAR values ​​were detected.

[0476] According to Table 9 of the experimental results showing changes in ADC DAR values, the complex formed with the small molecule connector of the present invention showed significantly smaller changes in DAR values ​​in human plasma compared to DS8201, demonstrating superior plasma stability and further illustrating the stability of the small molecule connector of the present invention. [Table 23]

[0477] Test Example 10: Her2-ADC Mouse PK Experiment

[0478] Test objectives

[0479] The pharmacokinetic properties of Her2-ADC in this application were evaluated using C57BL / 6J mice as test animals.

[0480] 1. Test drug ADC-5: 10 mg / kg ADC-9: 10 mg / kg ADC-10: 10 mg / kg ADC-13: 10 mg / kg

[0481] 2. Preparation method: All preparations were made by diluting with PBS.

[0482] 3. Test Method

[0483] Fifteen mice were administered the drug via tail vein, and plasma samples were collected from each mouse at a total of 10 time points: 0.083, 2, 8, 24, 48, 96, 168, 336, 504, and 672 hours. The concentrations of total antibody and ADC in the samples were measured by ELISA. The experimental results are shown in Table 10, and the ADC molecule of this application has good pharmacokinetic properties. [Table 24]

[0484] Test Example 11: Evaluation of drug efficacy in NCI-N87 tumor-carrying mice

[0485] Test objectives

[0486] The efficacy of Her2-ADC in this application was evaluated using Balb / c nude mice as test animals.

[0487] 1. Test drug ADC-3: 1.5 mg / kg ADC-5: 1.5 mg / kg ADC-10: 1.5 mg / kg Reference ADC (DS8201): 1.5 mg / kg Blank control: PBS

[0488] 2. Preparation method: All preparations were made by diluting with PBS.

[0489] 3. Test Method

[0490] NCI-N87 cells were subcutaneously inoculated into the right hypochondrium of mice, and tumors were allowed to grow for 7 days. The animals were then randomly divided into 5 groups of 6 mice each (4 experimental groups + 1 blank control group).

[0491] The drug was administered via tail vein injection, totaling one dose. Tumor volume and body weight were measured twice a week for four weeks, and the data were recorded. The data was statistically analyzed using Excel 2023 statistical software; the mean was calculated as avg, the SD value as STDEV, the SEM value as STDEV / SQRT, and the group difference P value as TTEST. The experimental results are shown in Table 11 and Figure 6. All of the ADC molecules in this application were able to significantly reduce tumor volume and had a considerable tumor-suppressing effect compared to the reference ADC. [Table 25]

[0492] Test Example 12: Evaluation of drug efficacy in JIMT-1 tumor-carrying mice

[0493] Test objectives

[0494] The efficacy of Her2-ADC in this application was evaluated using SCID Beige mice as test animals.

[0495] 1. Test drug ADC-3: 3 mg / kg ADC-5: 3 mg / kg ADC-9: 3 mg / kg ADC-10: 3 mg / kg ADC-13: 3 mg / kg ADC-14: 3 mg / kg Reference ADC (DS8201): 3 mg / kg Blank control: PBS

[0496] 2. Preparation method: All preparations were made by diluting with PBS.

[0497] 3. Test Method

[0498] JIMT-1 cells were subcutaneously inoculated into the right hypochondrium of mice, and tumors were allowed to grow for 8 days. The animals were then randomly divided into 8 groups of 6 mice each (7 experimental groups + 1 blank control group).

[0499] The drug was administered via tail vein injection, for a total of one dose. Tumor volume and body weight were measured twice a week for four weeks, and the data were recorded. The data was statistically analyzed using Excel 2023 statistical software; the mean was calculated as avg, the SD value as STDEV, the SEM value as STDEV / SQRT, and the group difference P value as TTEST. The experimental results are shown in Table 12 and Figure 7. All of the ADC molecules of this application were able to significantly reduce tumor volume and had a superior tumor suppression effect compared to the reference ADC. [Table 26]

[0500] The embodiments of the technical proposal of the present invention have been described exemplarily above. It should be understood that the scope of protection of the present invention is not limited to the embodiments described above. Any modifications, equivalent substitutions, improvements, etc., made by a person skilled in the art within the spirit and principles of the present invention should all be included within the scope of protection of the claims of this application.

Claims

1. A compound represented by formula I', its racemic mixture, stereoisomer, tautomer, isotopic marker, solvate, polytype, pharmaceutically acceptable salt, or prodrug compound thereof, 【Chemistry 1】 Here, R １ , R ２ , R ３ are the same or different and are, independently of each other, H, OH, CN, halogen, C １～１０ alkyl group, C ２～１０ alkenyl group, C ２～１０ alkynyl group, C １～１０ alkoxy group, halo C １～１０ alkyl group, halo C １～１０ alkoxy group, cyano C １～１０ alkyl group, cyano C １～１０ alkoxy group, C ３～１０ cycloalkyl group, and are selected from R ４ is H or 【Chemistry 2】 Selected from, R ４１ H, C １～６ alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5-14 membered heteroaryl groups, R ５ H, 【Transformation 3】 Selected from, R ５１ , R ５２ H and C are either the same or different, and are independent of each other. １～６ alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5-14 membered heteroaryl groups, R ５３ is C １～６ alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5- to 14-membered heteroaryl groups, the A ring is C ３～８ Cycloalkyl groups or C ３～８ Selected from heterocyclyl groups, Ra is H, hydroxyl group, CN, halogen, C １～６ alkyl group, C １～６ Selected from haloalkyl groups, n is selected from 0, 1, or 2, and q is selected from 0, 1, or 2. X is selected from CH or N, Y is - (CH ２ ) ｍ -O-(CH ２ ) ｐ - Selected from, m is selected from integers between 0 and 6. p is an integer selected from 0 to 6, and the compound is a racemate, stereoisomer, tautomer, isotopic marker, solvate, polytype, pharmaceutically acceptable salt, or prodrug compound thereof.

2. R １ H, OH, CN, halogen, C １～６ Alkyl alkyl group, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group or halo C １～６ Selected from alkoxy groups, Preferably, R １ The group is selected from H, OH, Br, methyl group, difluoromethoxy group, 2,2,2-trifluoroethoxy, vinyl group, cyclopropyl group, or ethynyl group. Preferably, R ２ is H, halogen, CN or C １～６ Selected from alkyl groups, Preferably, R ２ is selected from H or F, Preferably, R ３ is H or C １～６ Selected from alkyl groups, Preferably, R ３ The compound according to claim 1, wherein is H.

3. R ４ is H or 【Chemistry 4】 Selected from, for example 【Transformation 5】 And, Preferably, R ４ is H or 【Transformation 6】 Selected from, Preferably, X-R ４ teeth 【Transformation 7】 Preferably 【Transformation 8】 And, Preferably, X-R ４ ha-CH ２ - and Preferably, Y is -CH ２ -O-, and the compound according to claim 1 or 2, characterized in that.

4. R ５ H, 【Chemistry 9】 selected from, R ５１ is selected from H, a methyl group, an ethyl group, an isopropyl group or a cyclopropyl group, R ５２ is selected from H or a methyl group, R ５３ is selected from a methyl group, ring A is C ３～６ selected from a cycloalkyl group, Ra is H, a hydroxy group, CN, a halogen, C １～６ an alkyl group, C １～６ selected from a haloalkyl group, n is selected from 0 or 1, q is selected from 0 or 1, Preferably, the A ring is selected from a cyclobutane ring. Preferably, R ５ H, 【Chemistry 10】 Selected from, Preferably, R ５１ R is selected from H, methyl group, ethyl group, isopropyl group or cyclopropyl group, ５２ R is selected from H or a methyl group, ５３ The A ring is selected from a methyl group, and the A ring is selected from a cyclobutane ring. Preferably, R ５ H, 【Chemistry 11】 A compound according to any one of claims 1 to 3, characterized by being selected from among.

5. X is selected from CH or N, and if X is CH, R ４ If is H, or if X is N, then R ５ H is, Preferably, m is selected from 0, 1, or 2, and more preferably, m+p is 2, characterized in that the compound is according to any one of claims 1 to 4.

6. The structure of the compound shown in formula I' is as follows: 【Chemistry 12】 Here, R １ , R ２ , R ５１ , R ５２ The compound according to any one of claims 1 to 5, characterized in that n independently has the definition described in any one of claims 1 to 5.

7. The structure of compound I' is, 【Chemistry 13】 【Chemistry 14】 The compound according to any one of claims 1 to 6, characterized as shown above.

8. A compound represented by formula V, its racemic mixture, stereoisomer, tautomer, isotopically labeled, solvate, polytype, pharmaceutically acceptable salt, or prodrug compound thereof, M-L １ -L ２ -D (Form V) Here, M is the linker site with the antibody or its antigen-binding fragment. L １ is a peptide residue, preferably selected from glycine-glycine-phenylalanine-glycine (GGFG), glutamic acid-valine-citrulline (EVC), valine-citrulline (VC), aspartic acid-valine-citrulline (DVC), glutamic acid-glycine-glycine-phenylalanine-glycine (EGGFG), and aspartic acid-glycine-glycine-phenylalanine-glycine (DGGFG), preferably L １ It is selected from glycine-glycine-phenylalanine-glycine (GGFG), L ２ This is a linking group between the peptide residue and D, preferably a chemical bond, -NH-C １～６ Alkyl-, 【Chemistry 15】 Selected from, preferably L ２ NH-CH ２ - Selected from, D is a structural fragment of a biologically active molecule, and preferably, D is the structure of the compound shown in formula I after dehydrogenation. 【Chemistry 16】 Here, R １ , R ２ , R ３ They are the same or different, and independently of each other, H, OH, CN, halogens, C １～１０ Alkyl alkyl group, C ２～１０ Alkenyl group, C ２～１０ Alkynyl group, C １～１０ Alkoxy group, Halo C １～１０ Alkyl, Halo C １～１０ Alkoxy group, cyano C １～１０ Alkyl alkyl, cyano C １～１０ Alkoxy group, C ３～１０ Selected from cycloalkyl groups, R ４ is H or 【Chemistry 17】 Selected from, R ４１ H, C １～６ Alkyl alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ Alkyl alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5-14 membered heteroaryl groups, R ５ H, [Chemistry 18] Selected from, R ５１ , R ５２ H and C are either the same or different, and are independent of each other. １～６ Alkyl alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ Alkyl alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5-14 membered heteroaryl groups, R ５３ is C １～６ Alkyl alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ Alkyl alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5- to 14-membered heteroaryl groups, the A ring is C ３～８ Selected from cycloalkyl groups or 3- to 8-membered heterocyclyl groups, Ra is H, hydroxyl group, CN, halogen, C １～６ Alkyl alkyl group, C １～６ Selected from haloalkyl groups, n is selected from 0, 1, or 2, and q is selected from 0, 1, or 2. X is selected from CH or N, Y is - (CH ２ ) ｍ -, - (CH ２ ) ｍ -O-(CH ２ ) ｐ - Selected from, m is selected from integers between 0 and 6. p is selected from integers between 0 and 6. More preferably, the structure of D is as follows: 【Chemistry 19】 【Chemistry 20】 【Chemistry 21】 【Chemistry 22】 【Chemistry 23】 【Chemistry 24】 【Chemistry 25】 Preferably, M is selected from the following structures: 【Chemistry 26】 Lg is either absent or a leaving group, and the leaving group is selected from a halogen, a sulfonyl group, a trifluoromethanesulfonyl group, or a methylsulfonyl group, preferably Lg is a methylsulfonyl group. Ring B is selected from a 5-14 membered heteroaryl ring or a 3-14 membered heterocycle, preferably from a 5-6 membered N-containing heteroaryl ring or a 3-6 membered N-containing heterocycle. More preferably, the B ring is a pyrimidine ring, a pyridine ring, a triazine ring, or 【Chemistry 27】 Selected from, Each R ｂ They are either the same or different, and independently of each other are halogen, cyano group, oxo (=O), and C １～６ Alkyl, Halo C １～６ Alkyl alkyl groups, hydroxy C １～６ Alkyl alkyl group, C １～６ Alkoxy group or C ３～８ Cycloalkyl group, 3-8 membered heterocyclyl group, C １～６ Alkyl-O-C １～６ Alkyl-, C １～６ Selected from alkyl-(5-6 member) heteroaryl- groups, Preferably, each R ｂ These are either the same or different, and independently of each other are cyano, oxo (=O), methoxy, cyclopropyl, and trifluoromethyl groups. 【Chemistry 28】 Selected from, L ｍ１ It either does not exist, is unsubstituted, or has one, two, or more R values. ｍ１ C is optionally replaced by ６～１４ The groups are selected from aryl groups, 5-14 membered heteroaryl groups, and 3-14 membered heterocyclyl groups, and each R ｍ１ They are either the same or different, and independently of each other are H, halogen, cyano group, and C １～６ Alkyl alkyl, HOOC-C １～３ Selected from alkylene groups, Preferably, L ｍ１ It either does not exist, is unsubstituted, or has one, two, or more R values. ｍ１ The group that is optionally substituted is selected from a phenyl group, a piperidinyl group, or a piperazinyl group. more, L ｍ１ teeth, 【Chemistry 29】 Selected from, L ｍ２ This is either unsubstituted or one, two, or more Rs. ｍ２ It is optionally substituted by - (CH ２ ) ｓ -(C=O)-, -ethynyl-(CH ２ ) ｔ Selected from the base -(C=O)-, Preferably, L ｍ２ This is either unsubstituted or one, two, or more Rs. ｍ２ -CH is optionally substituted. ２ -(C=O)-, -(CH ２ ) ２ -(C=O)-, -(CH ２ ) ５ -(C=O)-, -ethynyl-(CH ２ ) ３ Selected from the base -(C=O)-, Each R ｍ２ They are either the same or different, and independently of each other are H, halogen, cyano group, and C １～６ Alkyl alkyl group or -C １～６ Selected from alkylene-COOH, the alkylene group is optionally interrupted by one, two, or more O, NH groups. Preferably, each R ｍ２ They are either the same or different, and are independent of each other -C １～３ Selected from alkylene-COOH, the alkylene group is optionally interrupted by O or NH. More specifically, R ｍ２ teeth, 【Transformation 30】 Selected from, r is selected from integers between 0 and 6. s is selected from integers between 0 and 6. t is selected from integers between 0 and 6. Preferably, M is selected from the following groups: 【Chemistry 31】 【Chemistry 32】 Preferably, M-L １ - L ２ teeth, 【Transformation 33】 【Transformation 34】 【Chemistry 35】 【Transformation 36】 【Chemistry 37】 Compounds, their racemates, stereoisomers, tautomers, isotopically labeled compounds, solvates, polytypes, pharmaceutically acceptable salts, or prodrug compounds thereof, selected from the above base.

9. The compound shown in formula V is 【Transformation 38】 【Chemistry 39】 【Chemistry 40】 【Chemistry 41】 【Chemistry 42】 【Chemistry 43】 【Chemistry 44】 【Chemistry 45】 【Chemistry 46】 【Chemistry 47】 【Chemistry 48】 【Chemistry 49】 [Transformation 50] 【Chemistry 51】 【Chemistry 52】 【Chemistry 53】 【Chemistry 54】 The compound according to claim 8, characterized by being selected from the following structure.

10. An antibody-drug conjugate having the structure shown in formula VI, A-[L-D] β (Form VI) Here, Ab is an antibody or its antigen-binding fragment, D has the definition described in claim 8 or 9, L is a connector linking Ab and D, and β is selected from an integer or decimal number from 1 to 10. Preferably, Ab is an antibody or an antigen-binding fragment, and the antigen-binding fragment is Fab, Fab', (Fab') ２ Fd, Fv, Fv linked by disulfide bonds, scFv, di-scFv, (scFv) ２ Selected from diabody and single-domain antibody (sdAb), and / or the antibody is a mouse-derived antibody, humanized antibody, chimeric antibody, bispecific antibody or multispecific antibody. Preferably, Ab is an anti-HER2 antibody or its antigen-binding fragment, and more preferably, Ab is trastuzumab or its antigen-binding fragment. Preferably, the antibody-drug conjugate is characterized in that β is selected from an integer or decimal number between 4 and 9 (for example, 7, 7.71, 7.84, 7.92, 7.94, 7.97, 7.98, 7.99, 8, 8.02, 8.06, or 8.14).

11. L is M'-L １ - L ２ Selected from, where M' is a linker site with an antibody or its antigen-binding fragment, and is formed by the complexation of M as defined in any one of claims 8 to 9 with an antibody or its antigen-binding fragment, and L １ , L ２ has the definition described in any one of claims 8 to 9, Preferably, M' is 【Transformation 55】 【Transformation 56】 Selected from, Preferably, the carbonyl group linking position in M' is L １ It is linked to, and the linking position in the heterocyclic or heteroaryl ring is linked to Ab, Preferably, L is 【Chemistry 57】 【Transformation 58】 【Chemistry 59】 【Transformation 60】 【Chemistry 61】 Selected from, Here, the first place is linked to Ab, and the second place is linked to D. Preferably, L-D is 【Transformation 62】 【Transformation 63】 【Chemistry 64】 【Transformation 65】 【Chemical Formula 66】 【Transformation 67】 【Transformation 68】 【Transformation 69】 【Transformation 70】 【Chemistry 71】 【Chemistry 72】 【Transformation 73】 【Chemistry 74】 【Chemistry 75】 【Transformation 76】 【Chemical 77】 【Transformation 78】 The antibody-drug conjugate according to claim 10, characterized by being selected from among.

12. The antibody-drug conjugate shown in formula VI is selected from the following structures: 【Transformation 79】 【Chemistry 80】 【Chemistry 81】 【Chemistry 82】 【Chemistry 83】 【Chemical 84】 【Chemical 85】 【Chemical 86】 The antibody-drug conjugate according to claim 10 or 11, characterized in that β is an integer or decimal between 7 and 9.

13. A pharmaceutical composition comprising at least one of the compounds or antibody-drug conjugates described in any one of claims 1 to 12, a racemate thereof, stereoisomers, tautomers, isotopic markers, solvates, polytypes, pharmaceutically acceptable salts thereof, or prodrug compounds thereof, in a therapeutically effective amount.

14. Use of at least one of the compounds or antibody-drug conjugates described in any one of claims 1 to 12, their racemates, stereoisomers, tautomers, isotopic markers, solvates, polytypes, pharmaceutically acceptable salts, or prodrug compounds thereof, or the pharmaceutical composition described in claim 14, in the manufacture of a topoisomerase I inhibitor and / or in the manufacture of a medicament for the prevention or treatment of a disease or disorder related to topoisomerase I, Preferably, the disease or disorder is a tumor, and the tumor includes breast cancer, gastric cancer, lung cancer, colorectal cancer, colon cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal cancer, melanoma, or leukemia.

15. A compound represented by formula I'', its racemic mixture, stereoisomer, tautomer, isotopic marker, solvate, polytype, pharmaceutically acceptable salt, or prodrug compound thereof, 【Chemistry 87】 Here, R １ , R ２ , R ３ They are the same or different, and independently of each other, H, OH, CN, halogens, C １～１０ Alkyl alkyl group, C ２～１０ Alkenyl group, C ２～１０ Alkynyl group, C １～１０ Alkoxy group, Halo C １～１０ Alkyl, Halo C １～１０ Alkoxy group, cyano C １～１０ Alkyl alkyl, cyano C １～１０ Alkoxy group, C ３～１０ Selected from cycloalkyl groups, R ４ is H or 【Chemical 88】 Selected from, R ４１ H, C １～６ Alkyl alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ Alkyl alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5-14 membered heteroaryl groups, R ５２ H, C １～６ Alkyl alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ Alkyl alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5-14 membered heteroaryl groups, R ２０ is selected from H or an amino protecting group, Preferably, R ２０ The following are selected from Fmoc, Boc, Bn, and Cbz: X is selected from CH or N, Y is - (CH ２ ) ｍ -O-(CH ２ ) ｐ - Selected from, m is selected from integers between 0 and 6. p is selected from integers between 0 and 6. Preferably, R １ H, OH, CN, halogen, C １～６ Alkyl alkyl group, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group or halo C １～６ Selected from alkoxy groups, Preferably, R １ The group is selected from H, OH, Br, methyl group, difluoromethoxy group, 2,2,2-trifluoroethoxy, vinyl group, cyclopropyl group, or ethynyl group. Preferably, R ２ is H, halogen, CN or C １～６ Selected from alkyl groups, Preferably, R ２ is selected from H or F, Preferably, R ３ is H or C １～６ Selected from alkyl groups, Preferably, R ３ H is, R ４ It is selected from H, Preferably, formula I'' is the following compound: 【Chemistry 89】 【Chemistry 90】 Compounds, their racemates, stereoisomers, tautomers, isotopic markers, solvates, polytypes, pharmaceutically acceptable salts, or prodrug compounds thereof.

16. A compound represented by formula MII, 【Chemistry 91】 Lg is a leaving group, and the leaving group is selected from a halogen, a sulfonyl group, a trifluoromethanesulfonyl group, or a methylsulfonyl group, preferably Lg is a methylsulfonyl group. Ring B is selected from a 5-6 membered N-containing heteroaryl ring or a 3-6 membered N-containing heteroring. More preferably, the B ring is selected from a pyrimidine ring, a pyridine ring, or a triazine ring. Each R ｂ They are either the same or different, and independently of each other are halogen, cyano group, oxo (=O), and C １～６ Alkyl, Halo C １～６ Alkyl alkyl groups, hydroxy C １～６ Alkyl alkyl group, C １～６ Alkoxy group or C ３～８ Cycloalkyl group, 3-8 membered heterocyclyl group, C １～６ Alkyl-O-C １～６ Alkyl-, C １～６ Selected from alkyl-(5-6 member) heteroaryl- groups, Preferably, each R ｂ These are either the same or different, and independently of each other are cyano, oxo (=O), methoxy, cyclopropyl, and trifluoromethyl groups. 【Chemistry 92】 Selected from, L ｍ１ It either does not exist, is unsubstituted, or has one, two, or more R values. ｍ１ C is optionally replaced by ６～１４ The groups are selected from aryl groups, 5-14 membered heteroaryl groups, and 3-14 membered heterocyclyl groups, and each R ｍ１ They are either the same or different, and independently of each other are H, halogen, cyano group, and C １～６ Alkyl alkyl, HOOC-C １～３ Selected from alkylene groups, Preferably, L ｍ１ It either does not exist, is unsubstituted, or has one, two, or more R values. ｍ１ The group that is optionally substituted is selected from a phenyl group, a piperidinyl group, or a piperazinyl group. more, L ｍ１ teeth, 【Chemistry 93】 Selected from, R ｍ２０ This is either unsubstituted or one, two, or more Rs. ｍ２ It is optionally substituted by - (CH ２ ) ｓ -(C=O)-R Ｚ ,-ethynyl-(CH ２ ) ｔ -(C=O)-R Ｚ Selected from the basis, Preferably, R ｍ２０ This is either unsubstituted or one, two, or more Rs. ｍ２ -CH is optionally substituted. ２ -(C=O)-R Ｚ ,-(CH ２ ) ２ -(C=O)-R Ｚ ,-(CH ２ ) ５ -(C=O)-R Ｚ ,-ethynyl-(CH ２ ) ３ -(C=O)-R Ｚ Selected from the basis, Each R ｍ２ They are either the same or different, and independently of each other are H, halogen, cyano group, and C １～６ Alkyl alkyl group or -C １～６ Selected from alkylene-COOH, the alkylene group is optionally interrupted by one, two, or more O, NH groups. Preferably, each R ｍ２ They are either the same or different, and are independent of each other -C １～３ Selected from alkylene-COOH, the alkylene group is optionally interrupted by O or NH. More specifically, R ｍ２ teeth, 【Chemical 94】 Selected from, r is selected from integers between 0 and 6. s is selected from integers between 0 and 6. t is selected from integers between 0 and 6. Rz is selected from a hydroxyl group, halogen, active ester, carboxyl protecting group, amino acid, peptide fragment, or hydrophilic fragment. The aforementioned amino acid is L １ The N-terminal amino acid is L, and the peptide fragment is L １ It is a subfragment formed by two, three, or four amino acids at the N-terminus of or L １ And, The C-terminus of the peptide fragment is a hydroxyl group, an active ester, a carboxyl protecting group, or 【Chemical 95】 And, Preferably, the hydrophilic fragment comprises a polyhydroxy group, a polyethylene glycol fragment, a betaine fragment, or a polycreatine fragment. Preferably, formula MII is formula MII-1 below, 【Chemistry 96】 Here, t and Rz are as defined above, and Z is N or CR ２２ And R ２１ and R ２２ These are H, halogen, cyano group, oxo (=O), and C, respectively, independently. １～６ Alkyl, Halo C １～６ Alkyl alkyl groups, hydroxy C １～６ Alkyl alkyl group, C １～６ Alkoxy group, C ３～８ Cycloalkyl group, 3-8 membered heterocyclyl group, C １～６ Alkyl-O-C １～６ Selected from alkyl groups, with the condition that Z is N, R ２１ It is not H, Preferably, Rz is OH or a halogen, t is an integer from 2 to 4, Z is N, and R ２１ C is a cyano group. １～６ Alkoxy group, C ３～４ Cycloalkyl groups or -C １～６ Alkyl-O-C １～６ Selected from alkyl groups, or R ２１ is H, and Z is CR ２２ And R ２２ is a cyano group or a trifluoromethyl group, Preferably, formula MII is, 【Chemistry 97】 This is a compound, a compound.

17. A compound represented by formula (L'-1)-(L'-3), 【Chem.98】 Lg is a leaving group, and the leaving group is selected from a halogen, a sulfonyl group, a trifluoromethanesulfonyl group, or a methylsulfonyl group, preferably Lg is a methylsulfonyl group. Ring B is selected from a 5-6 membered N-containing heteroaryl ring or a 3-6 membered N-containing heteroring. More preferably, the B ring is selected from a pyrimidine ring, a pyridine ring, or a triazine ring. Each R ｂ They are either the same or different, and independently of each other are halogen, cyano group, oxo (=O), and C １～６ Alkyl, Halo C １～６ Alkyl alkyl groups, hydroxy C １～６ Alkyl alkyl group, C １～６ Alkoxy group or C ３～８ Cycloalkyl group, 3-8 membered heterocyclyl group, C １～６ Alkyl-O-C １～６ Alkyl-, C １～６ Selected from alkyl-(5-6 member) heteroaryl- groups, Preferably, each R ｂ These are either the same or different, and independently of each other are cyano, oxo (=O), methoxy, cyclopropyl, and trifluoromethyl groups. 【Chem.99】 Selected from, L ｍ１ It either does not exist, is unsubstituted, or has one, two, or more R values. ｍ１ C is optionally replaced by ６～１４ The groups are selected from aryl groups, 5-14 membered heteroaryl groups, and 3-14 membered heterocyclyl groups, and each R ｍ１ They are either the same or different, and independently of each other are H, halogen, cyano group, and C １～６ Alkyl alkyl, HOOC-C １～３ Selected from alkylene groups, Preferably, L ｍ１ It either does not exist, is unsubstituted, or has one, two, or more R values. ｍ１ The group that is optionally substituted is selected from a phenyl group, a piperidinyl group, or a piperazinyl group. more, L ｍ１ teeth, 【Chemistry 100】 Selected from, L ｍ２ This is either unsubstituted or one, two, or more Rs. ｍ２ It is optionally substituted by - (CH ２ ) ｓ -(C=O)-, -ethynyl-(CH ２ ) ｔ Selected from the base -(C=O)-, Preferably, L ｍ２ This is either unsubstituted or one, two, or more Rs. ｍ２ -CH is optionally substituted. ２ -(C=O)-, -(CH ２ ) ２ -(C=O)-, -(CH ２ ) ５ -(C=O)-, -ethynyl-(CH ２ ) ３ Selected from the base -(C=O)-, Each R ｍ２ They are either the same or different, and independently of each other are H, halogen, cyano group, and C １～６ Alkyl alkyl group or -C １～６ Selected from alkylene-COOH, the alkylene group is optionally interrupted by one, two, or more O, NH groups. Preferably, each R ｍ２ They are either the same or different, and are independent of each other -C １～３ Selected from alkylene-COOH, the alkylene group is optionally interrupted by O or NH. More specifically, R ｍ２ teeth, 【Chemistry 101】 Selected from, r is selected from integers between 0 and 6. s is selected from integers between 0 and 6. t is selected from integers between 0 and 6. Rz ２ This is selected from a hydroxyl group, halogen, active ester, or carboxyl protecting group. Preferably, here, 【Chemical Engineering 102】 The fragment has the structure shown in the following formula, 【Chemistry 103】 Here, the definition of each group is as described in claim 16, R ５１ H and C are independent of each other. １～６ Alkyl alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２＋ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ Alkyl alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5-14 membered heteroaryl groups, R ５３ C １～６ Alkyl alkyl group, C １～６ Haloalkyl group, C １～６ Alkyl-NH-, (C １～６ Alkyl) ２ N-, C １～６ Alkyl-NH-C １～６ Alkyl alkyl group, (C １～６ Alkyl) ２ N-C １～６ Alkyl alkyl group, C １～６ Alkoxyalkyl groups, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3-6 membered heterocyclyl group, C ６～１４ Selected from aryl groups and 5-14 membered heteroaryl groups, the A ring is C ３～８ Selected from cycloalkyl groups or 3- to 8-membered heterocyclyl groups, Ra is H, hydroxyl group, CN, halogen, C １～６ Alkyl alkyl group, C １～６ Selected from haloalkyl groups, n is selected from 0, 1, or 2, and q is selected from 0, 1, or 2. L １ is a peptide residue, preferably selected from glycine-glycine-phenylalanine-glycine (GGFG), glutamic acid-valine-citrulline (EVC), valine-citrulline (VC), aspartic acid-valine-citrulline (DVC), glutamic acid-glycine-glycine-phenylalanine-glycine (EGGFG), and aspartic acid-glycine-glycine-phenylalanine-glycine (DGGFG), preferably L １ It is selected from glycine-glycine-phenylalanine-glycine (GGFG), L ２ This is a linking group between the peptide residue and D, preferably a chemical bond, -NH-C １～６ Alkyl-, 【Chemical 104】 Selected from, preferably L ２ NH-CH ２ - Selected from, (L'-1)-(L'-3) is preferably, 【Chemistry 105】 【Chemistry 106】 It is a compound.

18. A method for synthesizing formula (L') according to claim 17, wherein L ２ NH-CH ２ - and includes the following steps in the reaction equation: 【Chemistry 107】 Here, Rm ２０ The definition of is as described in claim 16, and Rz ２ A synthesis method wherein is a hydroxyl group, an active ester, or a carboxyl protecting group, and the definitions of the remaining groups are as described in claim 17.

19. A method for synthesizing the compound described in claim 8, comprising the first method or the second method, The first method is selected from the following steps: 【Chemistry 108】 Rz ２ The group is selected from a hydroxyl group, a halogen, an active ester, or a carboxyl protecting group, and the definitions of the remaining groups are as described in claim 8. The second method is, 【Chemistry 109】 A synthesis method comprising the step of reacting with a compound of formula I, wherein the compound of formula I is as described in claim 8, and the definitions of the remaining groups are as described in claim 8.

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