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

Abstract

The present invention relates to a camptothecin derivative represented by Formula I, a pharmaceutical composition, and a method for preparing and using the same. The camptothecin derivative has good tumor-inhibiting activity and can be used to treat or prevent tumors and to prepare medicines for treating or preventing neoplastic diseases. [Formula 1] JPEG2025541854000098.jpg49169

Description

Detailed Description of the Invention

[0001] This application claims priority to six applications, all of which are entitled "Camptothecin derivatives, pharmaceutical compositions, and methods for producing and using the same," including application number 202211739606.8 filed on December 30, 2022, application number 202310367781.7 filed on April 7, 2023, application number 202310743291.2 filed on June 21, 2023, application number 202310941713.7 filed on July 28, 2023, application number 202311173712.9 filed on September 12, 2023, and application number 202311802330.8 filed on December 25, 2023.

[0002] [Technical Field] The present invention belongs to the pharmaceutical field, specifically to camptothecin derivatives, pharmaceutical compositions and their preparation methods and uses.

[0003] [Background technology] Camptothecin (CPT) is a natural compound isolated from Camptotheca acuminata, a plant in the Botryllaceae family. Camptothecin is a five-ring fused compound consisting of a quinoline AB ring, a pyrrole C ring, a pyridinone D ring, and an α-hydroxylactone E ring, with the 20-position being S-configured (see the structural formula below). Due to its excellent anticancer activity, it was introduced into clinical practice in the early 1970s. However, clinical trials were subsequently discontinued due to the occurrence of serious side effects such as diarrhea and hemorrhagic cystitis. [ka]

[0004] Research data show that camptothecin forms a ternary complex with intracellular DNA topoisomerase I, inhibiting DNA unwinding and DNA replication, thereby causing cell death (Cancer Res. 1989, 49, 6365). Camptothecin and its derivatives have potent antitumor activity in animal in vivo models of lung cancer, breast cancer, colorectal cancer, and ovarian cancer (Nature Review Cancer. 2006, 6, 789). Currently, several camptothecin-like drugs are approved for the treatment of tumors (Med Res. Rev. 2015, 35, 753). Irinotecan is used to treat colorectal cancer, topotecan is used to treat ovarian cancer, and belotecan is used to treat ovarian cancer and small cell lung cancer. Camptothecin derivatives further include exatecan, rubitecan, diflomotecan, lutotecan, gimatecan, simmitecan, chimitecan, eromotecan, etc. Camptothecin drugs or derivatives often cause hematotoxicity due to bone marrow suppression, such as decreased white blood cells, decreased platelets, anemia, decreased neutrophils, etc., as well as gastrointestinal side effects, such as nausea, vomiting, and diarrhea. Clinical studies have found that ways to improve the safety and efficacy of camptothecin compounds include increasing their water solubility, improving their pharmacokinetic properties, improving activity, reducing dosage, or using their conjugates to form antibody-drug conjugates with antibodies. Therefore, there remains a high clinical need and application value for developing camptothecin compounds and their conjugates that have novel structures and can improve efficacy and ameliorate safety issues.

[0005] Summary of the Invention The present invention provides a compound of formula I, its racemate, stereoisomer, tautomer, isotopically labeled form, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound thereof, [ka] where R1, R2, and R3 are the same or different and independently represent H, OH, CN, halogen, C1~10 Alkyl group, C 2~10 Alkenyl group, C 2~10 Alkynyl group, C 1~10 Alkoxy group, haloC 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 groups, R4 is H or [ka] Selected from R 41 is H, C 1~6 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 groups, (C 1~6 Alkyl)2N-C 1~6 Alkyl group, C 1~6 Alkoxyalkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl groups, 3- to 6-membered heterocyclyl groups, C 6~14 selected from an aryl group and a 5- to 14-membered heteroaryl group; R5 is H, [ka] Selected from R 51 , R 52 are the same or different and are independently H, C 1~6 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 groups, (C 1~6 Alkyl)2N-C 1~6 Alkyl group, C 1~6Alkoxyalkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl groups, 3- to 6-membered heterocyclyl groups, C 6~14 R is selected from an aryl group and a 5- to 14-membered heteroaryl group; 53 is C 1~6 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 groups, (C 1~6 Alkyl)2N-C 1~6 Alkyl group, C 1~6 Alkoxyalkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl groups, 3- to 6-membered heterocyclyl groups, C 6~14 aryl group and 5- to 14-membered heteroaryl group, and ring A is C 3~8 cycloalkyl groups or 3- to 8-membered heterocyclyl groups, and Ra is H, a hydroxy group, CN, a halogen, or C 1~6 Alkyl group, C 1~6 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; m is selected from integers of 0 to 6.

[0006] According to some embodiments, R1 is H, OH, CN, halogen, C 1~6 Alkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cycloalkyl group or haloC 1~6 alkoxy groups, According to some embodiments, R1 is H, OH, CN, halogen, C 1~6 Alkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl or halo C 1~6 alkoxy groups, According to some embodiments, R1 is selected from H, OH, Br, methyl, difluoromethoxy, 2,2,2-trifluoroethoxy, vinyl, cyclopropyl, or ethynyl; 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, or an ethynyl group.

[0007] According to some embodiments, R2 is H, halogen, CN or C 1~6 alkyl groups, According to some embodiments, R2 is selected from H or F.

[0008] According to some embodiments, R3 is H or C 1~6 alkyl groups, According to some embodiments, R3 is H.

[0009] According to some embodiments, R4 is H or [ka] selected from, for example [ka] and According to some embodiments, R4 is H or [ka] is selected from.

[0010] According to some embodiments, the X-R4 [ka] and preferably [ka] is.

[0011] According to some embodiments, X-R4 is -CH2-.

[0012] According to some embodiments, R5 is H, [ka] Selected from R 51 is selected from H, methyl, ethyl, isopropyl, or cyclopropyl; R 52 is selected from H or a methyl group, and R 53 is selected from methyl groups, and the A ring is C 3~6 cycloalkyl groups, and Ra is H, a hydroxy group, CN, a halogen, or C 1~6 Alkyl group, C 1~6 haloalkyl groups, n is selected from 0 or 1, and q is selected from 0 or 1; According to some embodiments, the A ring is selected from a cyclobutane ring.

[0013] According to some embodiments, R5 is H, [ka] is selected from According to some embodiments, R 51 is selected from H, methyl, ethyl, isopropyl, or cyclopropyl; R 52 is selected from H or a methyl group, and R 53 is selected from a methyl group, and ring A is selected from a cyclobutane ring; According to some embodiments, R5 is H, [ka] is selected from.

[0014] According to some embodiments, R5 is H, [ka] is selected from.

[0015] According to some embodiments, X is selected from CH or N, and when X is CH, R4 is H, or when X is N, R5 is H; According to some embodiments, m is selected from 0, 1 or 2.

[0016] According to some embodiments, the structure of the compound of formula I is as shown below: [ka] R1, R2, R4, R5, X, and m independently have the definitions set forth herein.

[0017] According to some embodiments, the structure of the compound of formula I is as shown below: [ka] wherein R1, R2, and R5 independently have the definitions set forth herein.

[0018] According to some embodiments, the structure of the compound of formula I is as shown below: [ka] where R1, R2, R 51 , R 52 , the A ring, Ra, m, n, and q each independently have the definitions described herein.

[0019] According to an embodiment of the present invention, the structure of the compound of formula I is: [ka] JPEG2025541854000019.jpg195169 JPEG2025541854000020.jpg207169 JPEG2025541854000021.jpg244169 JPEG2025541854000022.jpg150169 As shown in the figure.

[0020] According to some embodiments, the present invention further provides a structural fragment D of the compound of formula I described herein after dehydrogenation, According to some embodiments, the structure of D is: [ka] JPEG2025541854000024.jpg190169 JPEG2025541854000025.jpg216169 JPEG2025541854000026.jpg192169 JPEG2025541854000027.jpg204169 As shown in the figure.

[0021] The present invention further provides a compound of formula V, its racemate, stereoisomer, tautomer, isotopically labeled, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound thereof: L'-D (Formula V) wherein L' is a linker containing a linker moiety M capable of reacting with an antibody or antigen-binding fragment thereof, and L'-D reacts with the antibody or antigen-binding fragment thereof, after which L' forms the linker L; Preferably, L' comprises a peptide residue L1 and a fragment L2 in which the peptide residue is attached to D.

[0022] The definition of D is as given herein.

[0023] The present invention further provides an antibody-drug conjugate according to Formula VI, Ab-[LD] β (Formula VI) Here, Ab is an antibody or an antigen-binding fragment thereof, D has the definition described herein, L is a linker connecting Ab and D, and β is selected from integers or decimals between 1 and 10.

[0024] The present invention further provides a method for preparing a compound of formula I, comprising the following method 1 or method 2: Method 1 is, Step (1) of removing the protecting group PG4 from compound I-41 to obtain compound I-42; and step (2) reacting compound I-42 with compound I-43 to obtain a compound of formula I, [ka] wherein R1, R2, R3, R4, R5, X, m have the definitions described herein, Y is a leaving group selected from, for example, OH, Cl, Br, I, and PG4 is an amino protecting group selected from, for example, Fmoc, Boc, Bn, Cbz.

[0025] Method 2 is Step (1) of removing the protecting group PG5 from compound I-51 to obtain compound I-52; and step (2) reacting compound I-52 with compound I-53 to obtain a compound of formula I, [ka]

[0026] where R1, R2, R3, R4, R5, R 51 , X, m, n have the definitions described herein, Y is a leaving group selected from, for example, OH, Cl, Br, I, and PG5 is an amino protecting group selected from, for example, Fmoc, Boc, Bn, Cbz.

[0027] The present invention further provides pharmaceutical compositions comprising a therapeutically effective amount of at least one of the compounds of Formula I, Formula V, their racemates, stereoisomers, tautomers, isotopically labeled forms, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof.

[0028] The present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of an antibody-drug conjugate shown in Formula VI.

[0029] According to an embodiment of the invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable auxiliary materials.

[0030] According to embodiments of the present invention, the pharmaceutical composition may further comprise one or more additional therapeutic agents.

[0031] The present invention further provides a method for treating a neoplastic disease, comprising administering to a patient a prophylactically or therapeutically effective amount of at least one of a compound represented by Formula I or Formula V or an antibody-drug conjugate represented by Formula VI, a racemate, stereoisomer, tautomer, isotopically labeled form, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound thereof.

[0032] The present invention further provides a method for treating a neoplastic disease, which comprises administering to a patient a prophylactically or therapeutically effective amount of the pharmaceutical composition described above.

[0033] The 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.

[0034] In some embodiments, the patient comprises a mammal, preferably a human.

[0035] The present invention further provides at least one of a compound represented by formula I, formula V, or an antibody-drug conjugate represented by formula VI, a racemate thereof, a stereoisomer, a tautomer, an isotope-labeled compound, a solvate, a polymorph, a pharmaceutically acceptable salt thereof, or a prodrug compound thereof, or a pharmaceutical composition thereof, for use in treating a neoplastic disease.

[0036] The present invention further provides the use of at least one of the compounds of Formula I, Formula V, or Formula VI, or their racemates, stereoisomers, tautomers, isotopically labeled forms, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof, or the pharmaceutical compositions described above, in the manufacture of a topoisomerase I inhibitor and / or in the manufacture of a medicament for preventing or treating a disease or disorder associated with topoisomerase I.

[0037] In some embodiments, the disease or disorder is a tumor, and the tumor comprises 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.

[0038] [Effects of the Invention] The compounds according to the present invention have good tumor-suppressing activity and can be used to manufacture medicaments for treating or preventing cancer (such as breast cancer or gastric cancer) and similar disorders and diseases.

[0039] Definitions and Explanations of Terms Unless otherwise stated, the radical and term definitions described in the specification and claims of this application, including their exemplary definitions, exemplary definitions, preferred definitions, definitions set forth in tables, definitions of specific compounds in the examples, etc., may be combined with each other in any combination, and it should be understood that such combined radical definitions and compound structures are within the scope of the description and / or claims of this application.

[0040] Unless otherwise specified, the numerical ranges described in this specification and claims are equivalent to describing at least each specific integer value therein. For example, the numerical range "1 to 12" is equivalent to describing 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 a specific numerical range is defined as a "number," it should be understood that both ends of the range, each integer within the range, and each decimal point within the range are described.

[0041] The term "halogen" means fluorine, chlorine, bromine and iodine.

[0042] "C 1~10 "Alkyl group" means straight and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, and "C 1-8 "Alkyl group" means a straight-chain or branched alkyl group having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. 1~6 The term "alkyl group" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, or 6 carbon atoms, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, or isomers thereof.

[0043] "C 2~10 An "alkenyl group" is to be understood to preferably 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, more preferably "C 2~8"C alkenyl group." 2~10 An "alkenyl group" includes one or more double bonds and has 2, 3, 4, 5, 6, 7, 8 carbon atoms, for example, 2, 3, 4, 5, or 6 carbon atoms (i.e., C 2~6 alkenyl groups), having 2 or 3 carbon atoms (i.e., C 2~3 It should be understood that the term "alkenyl group" preferably means a linear or branched monovalent hydrocarbon group. If the alkenyl group contains more than one double bond, the double bonds may be separate or conjugated. The alkenyl group may be, for example, a vinyl group, an allyl group, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z)-pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)-pent-1-enyl, (Z)-pent-1-enyl, hex-5-enyl, (E) -Hex-4-enyl, (Z)-hex-4-enyl, (E)-hex-3-enyl, (Z)-hex-3-enyl, (E)-hex-2-enyl, (Z)-hex-2-enyl, (E)-hex-1-enyl, (Z)-hex-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-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut (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-dimethylpropan-2-enyl, 1-ethylpropan-1-enyl, 1-propylvinyl, and 1-isopropylvinyl.

[0044] "C 2~10 An "alkynyl group" includes 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 alkynyl groups), having 2, 3, 4, 5, or 6 carbon atoms (i.e., "C 2~6 alkynyl groups), having 2 or 3 carbon atoms ("C 2~3 It is to be understood that the term "alkynyl group") preferably means a linear or branched monovalent hydrocarbon group. The alkynyl group is, for example, an ethynyl group, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-3-ynyl, 1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methyl pent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-3-ynyl, 1-ethylbut-2-ynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2,2-dimethylbut-3-ynyl, 1,1-dimethylbut-3-ynyl, 1,1-dimethylbut-2-ynyl or 3,3-dimethylbut-1-ynyl. In particular, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.

[0045] "C 3~10 The term "cycloalkyl group" should be understood to mean a saturated monovalent monocyclic, bicyclic (e.g., bridged, spiro) 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 cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl; or a bicyclic hydrocarbon group such as bornyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, dicyclo[2.1.1]hexyl, dicyclo[2.2.1]heptyl, dicyclo[2.2.1]heptenyl, 6,6-dimethyldicyclo[3.1.1]heptyl, 2,6,6-trimethyldicyclo[3.1.1]heptyl, dicyclo[2.2.2]octanyl, 2,7-diazaspiro[3.5]nonanyl, 2,6-diazaspiro[3.4]octanyl; or a tricyclic hydrocarbon group such as adamantyl.

[0046] Unless otherwise defined, the term "3- to 6-membered heterocyclyl group" refers to a saturated or unsaturated non-aromatic ring or ring system, e.g., it is a 4-, 5-, or 6-membered monocyclic ring and contains at least one, e.g., 1, 2, 3, 4, 5, or more, heteroatoms selected from O, S, and N, where N and S can be optionally oxidized to various oxidation states to form nitrogen oxides, -S(O)-, or -S(O)2-. The heterocyclyl group can also include fused or bridged rings and spirocycles. 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 pyrrolinyl 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.

[0047] "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 6~14aryl groups"), in particular rings having 6 carbon atoms ("C6 aryl groups"), such as phenyl or biphenyl groups, or rings having 9 carbon atoms ("C9 aryl groups"), such as indanyl or indenyl groups, or rings having 10 carbon atoms ("C 10 aryl group), such as a tetrahydronaphthyl group, a dihydronaphthyl group, or a naphthyl group, or a ring having 13 carbon atoms ("C 13 aryl group), such as 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. 6-20 When substituted, the aryl group may be mono- or polysubstituted, and the substitution position is not limited, and may be, for example, ortho-, para-, or meta-substituted.

[0048] The term "5- to 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, or 9, or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3, heteroatoms each independently selected from N, O, and S, and each of which may be benzo-fused. "Heteroaryl group" also refers to groups in which the heteroaromatic ring is fused to one or more aryl, alicyclic, or heterocyclyl rings, where the base or point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7- or 8-indolizinyl groups, 1-, 3-, 4-, 5-, 6- or 7-isoindolyl groups, 2-, 3-, 4-, 5-, 6- or 7-indolyl groups, 2-, 3-, 4-, 5-, 6- or 7-indazolyl groups, 2-, 4-, 5-, 6-, 7- or 8-purinyl groups, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-quinolidinyl groups, 2-, 3-, 4-, 5-, 6-, 7-, 8 ... 1-, 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-naphthyridinyl group, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl group, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl group, 2-, 4-, 6- or 7-pteridinyl group, 1-, 2 4aH Carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or 8-carbazolyl; Carbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-carbolinyl; 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenanthridinyl; 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-acridinyl; 1-, 2-, or 4- , 5-, 6-, 7-, 8- or 9-azinyl group, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthrolinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenazinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenoxazinyl group, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenazinyl 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-furo[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-furo[3,4-c]cinnolinyl, 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 7-benzoimidazolyl group, - or 7-benzothiazolyl group, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-benzoxapinyl group, 2-, 4-, 5-, 6-, 7- or 8-benzoazinyl group, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-4H-pyrrolo[1,2-b][2]benzazepinyl. Typical fused 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-benzoxazolyl 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 is bonded to another group to form a compound of the present invention, a carbon atom on the 5- to 20-membered heteroaryl ring may be bonded to the other group.A heteroatom on the 5- to 20-membered heteroaryl ring may be bonded to another group. When the 5- to 20-membered heteroaryl is substituted, it may be mono- or polysubstituted. The substitution position is not limited, and for example, a hydrogen atom bonded to a carbon atom on the heteroaryl ring may be substituted, or a hydrogen atom bonded to a heteroatom on the heteroaryl ring may be substituted.

[0049] The term "spirocycle" refers to a ring system in which two rings share one ring-forming atom.

[0050] The term "fused ring" refers to a ring system in which two rings share two ring-forming atoms.

[0051] The term "bridged ring" refers to a ring system in which two rings share three or more ring-forming atoms.

[0052] Unless otherwise specified, a heterocyclyl group, heteroaryl group, or heteroarylene group includes all possible isomeric forms thereof, for example, positional isomers thereof. Thus, some illustrative, non-limiting examples include those substituted at one, two, or more positions, such as the 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-position (if present), or those bonded to other groups, and include pyridin-2-yl, pyridin-2-ylidene, pyridin-3-yl, pyridin-3-ylidene, pyridin-4-yl, and pyridin-4-ylidene, thienyl or thienylene groups, including thiophen-2-yl, thiophen-2-ylidene, thiophen-3-yl, and thiophen-3-ylidene, pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

[0053] The term "alkyloxy group (alkoxy group)" refers to -O-(alkyl), where alkyl is as defined above. Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, and butoxy groups. An alkoxy group may be optionally substituted or unsubstituted; if substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkyloxy, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, and heterocycloalkyloxy groups.

[0054] The term "alkylamino group" refers to -NH-(alkyl group), where alkyl is as defined above. Non-limiting examples of alkylamino groups include methylamino, ethylamino, propylamino, isopropylamino, butylamino, and the like.

[0055] The term "(alkyl)amino" refers to -N-(alkyl), where alkyl is as defined above. Non-limiting examples of (alkyl)amino include dimethylamino, methylethylamino, diethylamino, dipropylamino, methylpropylamino, diisopropylamino, dibutylamino, and the like.

[0056] "Haloalkyl" refers to an alkyl group substituted with one or more halogens, where alkyl is defined above.

[0057] The peptide residue refers to an oligopeptide fragment in the ADC linker, which typically acts as a substrate for an enzyme in the tumor cell or microenvironment to control drug release, such as GGFG, and has the structure [ka] is.

[0058] As will be understood by those skilled in the art, the compounds of formula (I) may exist in various pharmaceutically acceptable salt forms: if they contain a basic center, they can form acid addition salts; if they contain 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 internal salts.

[0059] The compounds of the present invention can exist in the form of solvates (e.g., hydrates), in which the compounds of the present invention contain a polar solvent, in particular, for example, water, methanol, or ethanol, as a component of the compound's crystal lattice. The amount of polar solvent, in particular water, can be present in a stoichiometric or non-stoichiometric ratio.

[0060] Depending on their molecular structure, the compounds of the present invention may be chiral and therefore may exist in various enantiomeric forms. Therefore, these compounds can exist in racemic or optically active forms. The compounds of the present invention encompass isomers or mixtures thereof, racemates, in which each chiral carbon is in the R or S configuration. The compounds of the present invention or their intermediates can be isolated into enantiomeric 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 racemic amines, diastereomers were prepared from the mixture by reacting with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-benzenesulfonylproline), or various optically active camphorsulfonic acids. Chromatographic enantiomeric resolution can be advantageously carried out with optically active resolving agents (e.g., dinitrobenzoylphenylglycine, cellulose triacetate, or other carbohydrate derivatives, or chiral derivatized methacrylate polymers immobilized on silica gel). Suitable eluents for this purpose are water or alcohol-containing solvent mixtures, e.g., hexane / isopropanol / acetonitrile.

[0061] The corresponding stable isomers can be separated according to known methods, such as extraction, filtration or column chromatography.

[0062] The term "patient" refers to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse or primate, and most preferably a human.

[0063] The term "therapeutically effective amount" refers to an amount of an active compound or drug that elicits the biological or medical response a researcher, veterinarian, physician, or other clinician is looking for in a tissue, system, animal, individual, or human, including one or more of the following: (1) prevention of disease: e.g., preventing a disease, disorder, or disorder in an individual who is susceptible to the disease, disorder, or disorder but has not yet experienced or developed the pathology or symptoms of the disease; (2) inhibition of disease: e.g., inhibiting a disease, disorder, or disorder (i.e., preventing further progression of the pathology and / or symptoms) in an individual who is experiencing or has developed the pathology or symptoms of the disease, disorder, or disorder; or (3) amelioration of disease: e.g., ameliorating a disease, disorder, or disorder (i.e., reversing the pathology and / or symptoms) in an individual who is experiencing or has developed the pathology or symptoms of the disease, disorder, or disorder.

[0064] [Mode for Carrying Out the Invention] The following provides a more detailed description of the technical solutions of the present invention through specific examples. It should be understood that the following examples are merely for illustrative purposes and are not intended to limit the scope of protection of the present invention. Any technology realized based on the above content of the present invention is included within the intended scope of protection of the present invention.

[0065] Unless otherwise specified, all raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0066] 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 NMR data are expressed in ppm (ppm). A Bruker AVANCE-400 nuclear magnetic resonance spectrometer was used for the NMR measurements, and the solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0067] MS measurements were performed using an Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS liquid chromatograph mass spectrometer (manufacturer: Agilent, MS model number: 6110 / 6120 Quadrupole MS), a Waters ACQuity UPLC-QD / SQD (manufacturer: Waters, MS model number: Waters ACQuity Qda Detector / Waters SQ Detector), and a THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model number: THERMO Q Exactive).

[0068] For high performance liquid chromatography (HPLC) analysis, an Agilent 1260II HPLC and a Waters Acquity UPLC H-Class high performance liquid chromatograph were used.

[0069] Chiral HPLC analysis was performed using a Waters Acquity UPCC high performance liquid chromatograph.

[0070] The high-performance liquid preparative chromatographs used were 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.

[0071] CombiFlash flash chromatography was performed using Combiflash Rf200 (TELEDYNE ISCO).

[0072] The silica gel plates used for thin layer chromatography are Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plates. The silica gel plate specifications used for thin layer chromatography (TLC) are 0.15mm to 0.2mm, and the specifications of the products separated and purified by thin layer chromatography are 0.4mm to 0.5mm.

[0073] Silica gel column chromatography generally used Yantai Yellow Sea silica gel 200-300 mesh silica gel as the carrier.

[0074] Kinase mean inhibition rate and IC 50 The values ​​were measured using a NovoStar Microblade Reader (BMG GmbH, Germany).

[0075] Known starting materials of the present invention may be synthesized using or according to methods known in the art or may be purchased from companies such as ABCRGmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Shaoyuan Chemical Technology (Accela ChemBio Inc.), and Darui Chemical.

[0076] Unless otherwise stated in the examples, all reactions can be carried out under an argon or nitrogen atmosphere.

[0077] An argon or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1 L.

[0078] Hydrogen atmosphere refers to the reaction flask being connected to a hydrogen balloon with a volume of approximately 1 L.

[0079] The pressurized hydrogenation reaction was carried out using a Parr 3916EKX hydrogenation apparatus and a Seiran QL-500 hydrogen gas generator or an HC2-SS hydrogenation apparatus.

[0080] The hydrogenation reaction is usually carried out by evacuating the vessel and filling it with hydrogen gas, and this procedure is repeated three times.

[0081] The microwave reaction was carried out using a CEM Discover-S 908860 microwave reaction apparatus.

[0082] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0083] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0084] In the examples, the reaction process was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the eluent system of column chromatography used for purifying the compound, and the developing solvent system of thin layer chromatography comprised A: dichloromethane / methanol system and B: n-hexane / ethyl acetate system, and the volume ratio of the solvents was adjusted according to the polarity of the compound, optionally by adding small amounts of basic or acidic reagents such as triethylamine and acetic acid.

[0085] 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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0086] 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) and cooled to 0 °C. 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 h. After completion of the reaction, water (30 mL) was added to the reaction mixture, which was then extracted with dichloromethane (50 mL × 3). The combined organic phase was washed with saturated brine, dried, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 1b (20 g, yield: 84%).

[0087] MS m / z (ESI): 262.1 (M+H) + .

[0088] Step 2 (E)-4-(5-acetamido-3-fluoro-2-methoxyphenyl)but-3-enoic acid 1c Compound 1b (20 g, 76.3 mmol) was dissolved in dioxane (30 mL) and water (10 mL), and but-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 completion of the reaction, the reaction mixture was filtered, and the resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 1c (20 g, yield: 87%).

[0089] MS m / z (ESI): 268.1 (M+H) + .

[0090] Step 3 4-(5-acetamido-3-fluoro-2-methoxyphenyl)butyric acid 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 completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The resulting residue was purified by silica gel column chromatography using eluent system A to obtain the title compound 1d (20 g, yield: 99%).

[0091] MS m / z (ESI): 270.1 (M+1) + .

[0092] Step 4 N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 1e Compound 1d (20 g, 74.4 mmol) was dissolved in trifluoroacetic acid (60 mL) and cooled to 0 °C. Trifluoroacetic anhydride (31.24 g, 148.8 mmol) was then slowly added. The reaction mixture was stirred at room temperature for 7 hours. After completion of the reaction, the reaction mixture was slowly poured into water (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phase was washed with saturated aqueous sodium bicarbonate solution until neutral, then washed with saturated brine, dried, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 1e (9.2 g, yield: 46%).

[0093] MS m / z (ESI): 252.1 (M+1) + .

[0094] Step 5. (Z)-N-(3-fluoro-7-(hydroxyimino)-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-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 cooled 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 then 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 completion of the reaction, the reaction mixture was quenched with saturated ammonium chloride solution and extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with saturated brine, dried, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 1f (6 g, yield: 51%).

[0095] MS m / z (ESI): 281.1 (M+1) + .

[0096] Step 6: 1g of N-(7-amino-3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide (Z)-N-(3-Fluoro-7-(hydroxyimino)-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 1f (6 g, 21.4 mmol) was dissolved in dioxane (60 mL) and 2 N 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 completion of the reaction, the reaction mixture was filtered and concentrated to give the crude product 1g (5 g), which was used directly in the next step without further purification.

[0097] MS m / z (ESI): 267.1 (M+1) + .

[0098] Step 7 (9H-Fluoren-9-yl)methyl (8-acetamido-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 1h Compound 1g (5 g, 18.8 mmol) obtained in the previous step was dissolved in dioxane (50 mL) and adjusted to pH 7-8 with sodium bicarbonate. 9-Fluorenylmethyl-N-succinimidyl carbonate (6.36 g, 18.8 mmol) was then slowly added. The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was slowly poured into water (40 mL) and extracted with ethyl acetate (40 mL x 3). The combined organic phase was washed with saturated brine, dried, and concentrated.

[0099] The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 1h (5.3 g, yield: 54%).

[0100] MS m / z (ESI): 489.2 (M+1) + .

[0101] Step 8 (9H-Fluoren-9-yl)methyl (8-amino-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 1i Compound 1h (5 g, 10.25 mmol) was dissolved in dioxane (50 mL) and 12 N hydrochloric acid (10 mL). The reaction mixture was stirred at 60 °C for 2 hours. After completion of the reaction, the reaction mixture was slowly poured into water (50 mL) and extracted with ethyl acetate (40 mL × 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 1i (3.75 g, yield: 78%).

[0102] MS m / z (ESI): 447.2 (M+1) + .

[0103] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-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-pyranindolizine-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 h. After completion of the reaction, the mixture was extracted with ethyl acetate (30 mL × 3). The combined organic phase was washed with saturated brine, dried, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 1k (1.8 g, yield: 36%).

[0104] MS m / z (ESI): 674.2 (M+1) + .

[0105] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-1-yl)carbamate 1l Compound 1k (1.8 g, 2.7 mmol) was dissolved in 40% hydrobromic acid (40 mL). The reaction mixture was stirred at 100°C for 2 hours. After completion of the reaction, 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 by silica gel column chromatography using eluent system B to give the title compound 1l (1.08 g, yield: 60%).

[0106] MS m / z (ESI): 660.2 (M+1) + .

[0107] Step 11 (9S)-1-Amino-9-ethyl-5-fluoro-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4',6,7]indolizino[1,2-b]quinoline-10,13-dione 1m Compound 1l (500 mg, 0.7 mmol) was dissolved in N,N-dimethylformamide (5 mL) and diethylamine (166 mg, 2.3 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the diethylamine in the reaction mixture was removed by rotary drying using an oil pump. The crude product 1m obtained by rotary drying was triturated with ethyl acetate, and the resulting solid was directly added to the next step of the reaction.

[0108] MS m / z (ESI): 438.1 (M+1) + .

[0109] 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]indolizino[1,2-b]quinolin-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 completion of the reaction, the reaction mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phase was washed with saturated brine, dried, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 1n (90 mg, yield: 64%).

[0110] MS m / z (ESI): 610.2 (M+1) + .

[0111] 13th step (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide Compound 1n (90 mg, 0.15 mmol) was dissolved in methanol (5 mL) and 1 M aqueous lithium hydroxide solution (2 mL) was added at room temperature. The reaction mixture was stirred at room temperature for 15 minutes. After completion of the reaction, the methanol was rotary evaporated, and the remaining aqueous phase was lyophilized 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; 15-minute gradient, gradient ratio: acetonitrile phase 23% to 33%, flow rate: 20 mL / min) to obtain Compound 1-1 (35 mg, yield: 30%) and Compound 1-2 (28 mg, yield: 24%).

[0112] Single configuration compound 1-1 (shorter retention time): MS m / z (ESI): 524.2 (M+1) + .

[0113] 1H NMR (400MHz, DMSO-d6) δ 10.42 (s, 1H), 8.39 (d, 1H), 7.84 (d, 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, 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, 3H), 0.93 - 0.81 (m, 3H).

[0114] Single-configuration compound 1-2 (long retention time): MS m / z (ESI): 524.2 (M+1) + .

[0115] 1 H NMR (400MHz, DMSO-d6) δ 10.42 (s, 1H), 8.42 (d, 1H), 7.84 (d, 1H), 7.26 (s, 1H), 5.59 - 5.51 (m, 1H), 5.42 (s, 2H), 5.29 - 5.14 (m, 2H), 4.04 (dd, 1H), 3.17 (dd, 1H), 3.10 - 2.92 (m, 1H), 2.28 (dd, 1H), 2.18 (dd, 1H), 2.13 - 1.95 (m, 2H), 1.93 - 1.78 (m, 2H), 1.08 (d, 3H), 0.91 - 0.82 (m, 3H).

[0116] 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]indolizine[1,2-b]quinolin-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]indolizine[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0117] Using the synthetic route of Example 1, the raw material in Step 1 was replaced with 3-bromo-4-methoxyaniline 2a (25 g, 0.12 mol). The resulting 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; 15-minute gradient, gradient ratio: acetonitrile phase 23% to 33%, flow rate: 20 mL / min) to give the title products 2-1 (19 mg, yield: 28%) and 2-2 (30 mg, yield: 44%).

[0118] Single-configuration compound 2-1 (short retention time): MS m / z (ESI): 506 (M+1) + .

[0119] 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).

[0120] Single stereo compound 2-2 (long retention time): MS m / z (ESI): 506 (M+1) + .

[0121] 1 H NMR (400MHz, DMSO-d6) δ 10.22 (s, 1H), 8.42 - 8.40 (d, 1H), 7.93 - 7.90 (d, 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).

[0122] 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]indolizine[1,2-b]quinolin-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]indolizine[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0123] 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 completed, 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 rotary evaporated to give crude product 3b (66 g).

[0124] MS m / z (ESI): 315.9, 317.9 (M+1) + .

[0125] 1 H NMR (400MHz, DMSO-d6) δ 7.01 (d, 1H), 6.47 (dd, 1H), 5.74 (s, 2H).

[0126] 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 mixture to 0 °C, acetyl chloride (19.68 g, 250.70 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature for 4 h. After the reaction was complete, the solvent was rotary evaporated to obtain the crude product. The crude product was redissolved in ethyl acetate and the pH of the mixture was adjusted to 2-3 with dilute hydrochloric acid. The mixture was extracted with ethyl acetate, washed with saturated brine, and the organic phase was collected and dried over anhydrous sodium sulfate. The solvent was rotary evaporated to obtain the crude product. The crude product was triturated with a dichloromethane / methanol (10:1) mixture to obtain the title compound 3c (60 g, yield: 80%).

[0127] MS m / z (ESI): 357.9, 359.9 (M+1) + .

[0128] 1 H NMR (400MHz, DMSO-d6) δ 10.27 (s, 1H), 7.96 (s, 1H), 7.66 (dd, 1H), 2.06 (s, 3H).

[0129] Step 3 (E)-4-(5-acetamido-2-bromo-3-fluorophenyl)but-3-enoic acid 3d Compound 3c (20 g, 55.87 mmol) was dissolved in a mixture of dioxane (200 mL) and water (40 mL), and but-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. Under nitrogen gas protection, the reaction mixture was stirred at 100 °C for 16 h. After completion of the reaction, water and dichloromethane were added, and the mixture was washed with saturated sodium bicarbonate solution 3-5 times. The aqueous phase was collected. The pH of the aqueous phase was adjusted to 2-3 with hydrochloric acid and extracted 5-7 times with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was rotary evaporated to give crude product compound 3d (17 g).

[0130] MS m / z (ESI): 316.0, 317.9 (M+1) + .

[0131] Step 4: 4-(5-acetamido-2-bromo-3-fluorophenyl)butyric acid 3e Compound 3d (8 g, 25.3 mmol) was dissolved in methanol (80 mL) and platinum-on-carbon catalyst (800 mg) was added. The reaction was stirred at room temperature under hydrogen gas for 2 hours. After completion of the reaction, the mixture was filtered, the filtrate was collected, and the solvent was evaporated by rotary evaporation to give crude product 3e (8 g). The crude product was used directly in the next step without further purification.

[0132] MS m / z (ESI): 318.0, 320.0 (M+1) + .

[0133] 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).

[0134] Step 5. N-(4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 3f Compound 3e (8 g, 25.1 mmol) was dissolved in trifluoroacetic acid (80 mL), and trifluoroacetic anhydride (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 completed, water was added under ice bath conditions, and the pH of the system was adjusted to 9-10 with 15% sodium hydroxide solution. The system was then extracted with dichloromethane, and the organic phase was washed with saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to give compound 3f (4.8 g, yield: 53%).

[0135] MS m / z (ESI): 329.0, 331.0 (M+1) + .

[0136] Step 6: 3g of (Z)-N-(4-bromo-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide Potassium tert-butoxide (1.62 g, 14.4 mmol) was dissolved in a mixture of tetrahydrofuran (80 mL) and tert-butanol (20 mL). Compound 3f (2.15 g, 7.2 mmol) in tetrahydrofuran (20 mL) was slowly added in an ice bath. The reaction mixture was stirred at 0 °C for 10 min, after which isoamyl nitrite (1.27 g, 10.8 mmol) was added, and the mixture was stirred at 0 °C for 50 min. After the reaction was complete, the pH of the mixture was adjusted to 4-5 with dilute hydrochloric acid, extracted with ethyl acetate, and the organic phase was washed with saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was rotary evaporated to give the crude product. The crude product was triturated with methyl tert-butyl ether to give the title compound 3g (1.1 g, yield: 46%).

[0137] MS m / z (ESI): 329.0, 331.0 (M+1) + .

[0138] 1 H NMR (400MHz, CD3OD) δ 8.52 (d, 1H), 3.21 (dd, 2H), 3.07 (dd, 2H), 2.24 (s, 3H).

[0139] Step 7 N-(7-amino-4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 3h Compound 3g (500 mg, 1.52 mmol) was dissolved in dioxane (10 mL), and 1 mL of 1 M hydrochloric acid and platinum-on-carbon catalyst (100 mg) were added. The reaction was stirred at room temperature under a hydrogen atmosphere for 4 hours. After completion of the reaction, the mixture was filtered, and the filtrate was collected and concentrated to give crude product 3h (500 mg), which was used directly in the next step.

[0140] MS m / z (ESI): 315.1, 317.1 (M+1) + .

[0141] Step 8 (9H-Fluoren-9-yl)methyl (8-acetamido-5-bromo-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 3i Compound 3h (500 mg) was dissolved in dioxane (10 mL). The pH of the filtrate from step 7 was adjusted to 8-9 with saturated sodium carbonate solution, and then fluorenylmethoxycarbonyl chloride (432 mg, 1.67 mmol) was added. The reaction was stirred at room temperature for 1 hour. After the reaction was completed, the system was extracted with ethyl acetate, and the organic phase was washed with saturated brine. The organic phase was collected and dried over anhydrous sodium sulfate. The solvent was then rotary evaporated to give the crude product. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 3i (300 mg, yield: 37%).

[0142] MS m / z (ESI): 537.0, 539.0 (M+1) + .

[0143] Step 9 (9H-Fluoren-9-yl)methyl (8-amino-5-bromo-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-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 completion of the reaction, the resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 3j (500 mg, yield: 77%).

[0144] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-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-pyrano[3,4-f]indolizine-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 mixture was stirred at 120 °C for 2 h. After completion of the reaction, the solvent was rotary evaporated to give the crude product. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 3k (250 mg, yield: 86%).

[0145] MS m / z (ESI): 722.0, 724.0 (M+1) + .

[0146] 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).

[0147] Step 11 (9S)-1-Amino-4-bromo-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4',6,7]indolizino[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 completion of the reaction, the resulting residue was purified by silica gel column chromatography using eluent system A to give the title compound 3l (100 mg, yield: 72%).

[0148] MS m / z (ESI): 500.0, 502.1 (M+1) + .

[0149] 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]indolizine[1,2-b]quinolin-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]indolizine[1,2-b]quinolin-1-yl)-3-hydroxybutanamide Compound 3l (100 mg, 0.20 mmol) was dissolved in DMF (2 mL), 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, and after completion of the reaction, the residue 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 (0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 38% to 48%, flow rate: 25 mL / min) to give compound 3-1 (1.62 mg, yield: 6%) and compound 3-2 (1.32 mg, yield: 7.4%).

[0150] Single-configuration compound 3-1 (short retention time): MS m / z (ESI): 586.0, 588.0 (M+1) + .

[0151] 1 H NMR (400MHz, CD3OD) δ 8.10 (d, 1H), 7.90 (s, 1H), 5.96 - 5.88 (m, 1H), 5.83 (d, 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, 3H), 1.25 (t, 3H).

[0152] Single-configuration compound 3-2 (long retention time): MS m / z (ESI): 586.0, 588.0 (M+1) + .

[0153] 1H NMR (400MHz, CD3OD) δ 7.79 (d, 1H), 7.59 (s, 1H), 5.67 - 5.59 (m, 1H), 5.52 (d, 1H), 5.41 - 5.33 (m, 2H), 5.32 - 5.23 (m, 2H), 4.24 - 4.18 (m, 1H), 2.34 (d, 2H), 2.28 - 2.21 (m, 2H), 1.93 - 1.86 (m, 3H), 1.17 (d, 3H), 0.94 (t, 3H).

[0154] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0155] Using the synthetic 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; 15-minute gradient, gradient blend ratio: acetonitrile phase 38% to 48%, flow rate: 25 mL / min) to give the title products 4-1 (2.5 mg, yield: 12%) and 4-2 (3.1 mg, yield: 15%).

[0156] Single-configuration compound 4-1 (short retention time): MS m / z (ESI): 508.1 (M+1) + .

[0157] 1 H NMR (400MHz, CD3OD) δ 7.65 (d, 2H), 7.37 (d, 1H), 5.66 (s, 1H), 5.56 (d, 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, 4H), 0.99 (t, 3H).

[0158] Single-configuration compound 4-2 (long retention time): MS m / z (ESI): 508.1 (M+1) + .

[0159] 1 H NMR (400MHz, CD3OD) δ 7.57 (d, 2H), 7.30 (d, 1H), 5.60 (d, 1H), 5.48 (d, 1H), 5.36 - 5.15 (m, 5H), 4.22 - 4.15 (m, 1H), 2.36 - 2.30 (m, 2H), 2.18 (t, 3H), 1.90 - 1.83 (m, 2H), 1.15 (d, 3H), 0.91 (t, 3H).

[0160] 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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0161] First step: (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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-1-yl)carbamate 5a Compound 1l (200 mg, 0.30 mmol) was dissolved in DMF (5 mL), and cesium carbonate (198 mg, 0.61 mmol) and (bromodifluoromethyl)phosphonic acid diethyl ester (121 mg, 0.45 mmol) were added in an ice bath. The reaction mixture was stirred at 0°C for 1 hour. After the reaction was completed, water was added, the mixture was extracted with ethyl acetate, and the organic phase was washed with saturated brine. The organic phase was collected and dried over anhydrous sodium sulfate. The solvent was then rotary evaporated to obtain the crude product. The crude product was purified by column chromatography to obtain the title compound 5a (200 mg, yield: 93%).

[0162] MS m / z (ESI): 711.2 (M+1) + .

[0163] Step 2 (9S)-1-Amino-4-(difluoromethoxy)-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4',6,7]indolizino[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 completion of the reaction, the reaction solution was rotary evaporated under vacuum using an oil pump to obtain the crude product 5b (90 mg). The crude product was used directly in the next step without further purification.

[0164] MS m / z (ESI): 488.1 (M+1) + .

[0165] 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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide The 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 completion of the reaction, the solvent was rotary evaporated to obtain a crude product, which was purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, chromatography column: Xtimate C18 250*30 mm, 10 μm; mobile phase 1: water (0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 37% to 47%, flow rate: 50 mL / min) to obtain the title compound 5-1 (8.6 mg, yield: 16%) and the title compound 5-2 (7.8 mg, yield: 14.7%).

[0166] Single-configuration compound 5-1 (short retention time): MS m / z (ESI): 574.1 (M+1) + .

[0167] 1 H NMR (400MHz, DMSO-d6) δ 8.49 (d, 2H), 8.06 (d, 1H), 7.34 (s, 1H),7.31 (t, 1H) 6.54 (s, 1H), 5.63 - 5.54 (m, 1H), 5.43 (s, 2H), 5.32 - 5.16 (m, 2H), 4.68 (d, 1H), 4.05 (s, 1H), 3.23 (d, 1H), 2.36 - 2.19 (m, 2H), 2.19 - 2.05 (m, 2H), 1.93 - 1.78 (m, 2H), 1.10 (d, 3H), 0.88 (t, 3H).

[0168] Single-configuration compound 5-2 (long retention time): MS m / z (ESI): 574.1 (M+1)+ .

[0169] 1 H NMR (400MHz, DMSO-d6) δ 8.52 (d, 2H), 8.10 - 8.04 (m, 1H), 7.35 (s, 1H), 7.32 (t, 1 H) 6.55 (s, 1H), 5.66 - 5.58 (m, 1H), 5.43 (d, 2H), 5.26 (t, 2H), 4.66 (d, 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, 3H), 0.89 (t, 3H).

[0170] 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-cycloheptane[3',4',6,7]indolizino[1,2-b]quinolin-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-cycloheptane[3',4',6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0171] 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 portionwise at that temperature, and the reaction was continued for 2 hours while maintaining the temperature at 60 °C. The reaction mixture was cooled to room temperature and added dropwise to ice water, extracted with toluene, and the combined organic phases were washed successively with sodium sulfite solution, water, and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the title compound 6b (30.0 g). The product was used directly in the next step without further purification.

[0172] MS m / z (ESI): 233.9 (M+1) + .

[0173] 1 H NMR (400MHz, CDCl3) δ 8.29-8.23 (m, 1H), 7.88 (dd, 1H), 2.44 (d, 3H).

[0174] Step 2 3-Bromo-5-fluoro-4-methylaniline 6c Compound 6b (30.0 g, 0.13 mol) was dissolved in methanol (200 mL), and platinum on carbon (3.0 g, 5% content) was added. After replacing the atmosphere with hydrogen gas, the mixture was reacted at room temperature for 16 hours under a hydrogen gas atmosphere. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated to give title compound 6c (20.0 g, yellow oil). This product was used directly in the next step without further purification.

[0175] MS m / z (ESI): 204.0 (M+1) + .

[0176] Step 3 N-(3-bromo-5-fluoro-4-methylphenyl)acetamide 6d 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 allowed to react for 3 hours under ice-bath conditions. After completion of the reaction, the reaction mixture was quenched by adding water, extracted with ethyl acetate, and the combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product. The crude product was triturated with a mixture of dichloromethane and petroleum ether (V / V = 10:1) to give the title compound 6d (10.0 g, yield: 32%).

[0177] MS m / z (ESI): 246.0 (M+1) + .

[0178] Step 4 (E)-5-(5-acetamido-3-fluoro-2-methylphenyl)pent-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 pent-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 h. After completion of the reaction, water and dichloromethane were added, and the mixture was washed three times with saturated sodium bicarbonate solution. The aqueous phase was collected. The pH of the aqueous phase was adjusted to 2-3 with hydrochloric acid and extracted three times with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was rotary evaporated to give crude product compound 6e (10 g). The product was directly used in the next step reaction without purification.

[0179] MS m / z (ESI): 266.1 (M+1) + .

[0180] The subsequent synthesis route was the same as in Example 3, except that intermediate 3d in Example 3 was replaced 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, chromatography column: Xbridge C18 150*19 mm, 5 μm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; 15 min gradient, gradient ratio: acetonitrile phase 37% to 47%, flow rate: 20 mL / min) to give the title products 6-1 (12.6 mg, yield: 14%) and 6-2 (17.2 mg, yield: 19%).

[0181] Single-configuration compound 6-1 (short retention time): MS m / z (ESI): 536 (M+1) + .

[0182] 1 H NMR (400MHz, 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, 4H), 1.06 (d, 3H), 0.85 - 0.82 (m, 3H).

[0183] Single-configuration compound 6-2 (long retention time): MS m / z (ESI): 536 (M+1) + .

[0184] 1H NMR (400MHz, 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, 3H), 2.40 - 2.15 (m, 4H), 2.11 - 1.52 (m, 4H), 1.08 - 1.07 (m, 3H), 0.87 - 0.85 (m, 3H).

[0185] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0186] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-1-yl)carbamate 7a Compound 3k (140 mg, 0.20 mmol) and potassium vinyltrifluoroborate (54 mg, 0.40 mmol) were dissolved in dioxane (16 mL) and water (4 mL). Potassium phosphate (128 mg, 0.60 mmol) and (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (34 mg, 0.04 mmol) were added, and the atmosphere was purged with nitrogen gas. The reaction mixture was stirred at 100 °C for 5 hours. After completion of the reaction, water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The combined organic phase was washed with saturated brine, dried, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent system B to give the title compound 7a (80 mg, yield: 60%).

[0187] MS m / z (ESI): 670.2 (M+1) + .

[0188] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinoline-10,13-dione Compound 7a (80 mg, 0.12 mmol) was dissolved in N,N-dimethylformamide (10 mL), and diethylamine (1 mL) was added to the reaction mixture, followed by stirring at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated in vacuo, triturated with ethyl acetate, and filtered to give the title compound 7b (30 mg, yield: 51%).

[0189] MS m / z (ESI): 448.2 (M+1) + .

[0190] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-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 h. After completion of the reaction, the reaction solution was concentrated and 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% formic acid); mobile phase 2: acetonitrile; 15-minute gradient, gradient blending ratio: acetonitrile phase 43% to 53%, flow rate: 20 mL / min) to give the title compound 7-1 (1.76 mg, yield: 10%) and the title compound 7-2 (2.31 mg, yield: 10%).

[0191] Single-configuration compound 7-1 (short retention time): MS m / z (ESI): 534.2 (M+1) + .

[0192] 1H NMR (400MHz, DMSO-d6) δ 8.51 (d, 1H), 8.48 (s, 1H), 7.84 (d, 1H), 7.32 (s, 1H), 6.97 (dd, 1H), 6.58 (s, 1H), 5.91 - 5.77 (m, 2H), 5.61 - 5.50 (m, 1H), 5.43 (s, 2H), 5.22 (q, 2H), 4.72 (s, 1H), 4.04 (dd, 1H), 3.27 (d, 1H), 2.30 - 2.20 (m, 2H), 2.11 (t, 2H), 1.90 - 1.80 (m, 2H), 1.09 (d, 3H), 0.87 (t, 3H).

[0193] Single-configuration compound 7-2 (long retention time): MS m / z (ESI): 534.2 (M+1) + .

[0194] 1 H NMR (400MHz, DMSO-d6) δ 8.55 (d, 1H), 8.47 (s, 1H), 7.85 (d, 1H), 7.32 (s, 1H), 6.97 (dd, 1H), 6.62 (s, 1H), 5.82 (dd, 2H), 5.57 (dt, 1H), 5.43 (d, 2H), 5.24 (s, 2H), 4.72 (s, 1H), 4.03 (dt, 1H), 3.27 (d, 1H), 2.23 (ddd, 2H), 2.08 (dd, 2H), 1.87 (tt, 2H), 1.08 (d, 3H), 0.87 (s, 3H).

[0195] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0196] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-1-yl)carbamate 8a Compound 1l (200 mg, 0.30 mmol) was dissolved in N,N-dimethylformamide (3 mL), and 2,2,2-trifluoroethyl trifluoromethanesulfonate (141 mg, 0.60 mmol) and cesium carbonate (296 mg, 0.90 mmol) were slowly added. The reaction mixture was stirred at room temperature for 10 minutes. After the reaction was complete, the mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL × 3). The organic phases were combined and concentrated. The residue was purified by silica gel column chromatography using eluent system B to give the title compound 8a (180 mg, yield: 75%).

[0197] MS m / z (ESI): 742.2 (M+H) + .

[0198] 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]pyrano[3',4',6,7]indolizino[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 completion of the reaction, the reaction mixture was concentrated under reduced pressure to give crude product 8b (100 mg). This product was used directly in the next step without further purification.

[0199] MS m / z (ESI): 520.1 (M+H) + .

[0200] 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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-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]pyrano[3',4',6,7]indolizino[1,2-b]quinolin-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 completion of the reaction, the mixture was concentrated under reduced pressure to obtain a crude product, which was then purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, column: Xtimate C18 150*19 mm, 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 38% to 48%, flow rate: 50 mL / min) to obtain the title compound 8-1 (2.5 mg, yield: 4.6%) and the title compound 8-2 (3.5 mg, yield: 6.4%).

[0201] Single-configuration compound 8-1 (shorter retention time): MS m / z (ESI): 606.2 (M+1) + .

[0202] 1 H NMR (400MHz, DMSO-d6) δ 8.43 (d, 1H), 7.98 (d, 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, 1H), 4.08 - 4.03 (m, 1H), 3.21 (d, 2H), 2.34 - 2.27 (m, 1H), 2.22 (d, 1H), 2.10 (dd, 2H), 1.89 - 1.81 (m, 2H), 1.10 (d, 3H), 0.90 - 0.84 (m, 3H).

[0203] Single-configuration compound 8-2 (long retention time): MS m / z (ESI): 606.2 (M+1)+ .

[0204] 1 H NMR (400MHz, DMSO-d6) δ 8.45 (d, 1H), 7.98 (d, 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, 2H), 4.65 (d, 1H), 4.09 - 4.00 (m, 1H), 3.24 - 3.12 (m, 2H), 2.28 (dd, 1H), 2.19 (dd, 1H), 2.09 (dd, 2H), 1.93 - 1.78 (m, 2H), 1.09 (d, 3H), 0.94 - 0.78 (m, 3H).

[0205] Example 9 (S)-9-Ethyl-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzopyrano[3',4',6,7]indolizino[1,2-b]quinoline-10,13-dione 9 [ka]

[0206] Step 1: 8-Amino-5-methoxy-3,4-dihydronaphthalen-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 reacted at 60 °C for 2 hours. After completion of the reaction, the reaction mixture was directly concentrated to give the title compound 9a (400 mg, yield: 88%).

[0207] MS m / z (ESI): 192.1 (M+1) + .

[0208] Step 2 (S)-9-Ethyl-9-hydroxy-4-methoxy-1,2,3,9,12,15-hexahydro-10H,13H-benzopyrano[3',4',6,7]indolizino[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 completion of the reaction, water (20 mL) was added to the reaction mixture, which was then extracted with dichloromethane (15 mL × 3). The combined organic phase was washed with saturated brine, dried, and concentrated. The mixture was purified by silica gel column chromatography using eluent system B to obtain the title compound 9b (698 mg, yield: 80%).

[0209] MS m / z (ESI): 419.1 (M+1) + .

[0210] Step 3 (S)-9-Ethyl-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzopyrano[3',4',6,7]indolizino[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 incubated at 100°C for 2 hours. After completion of the reaction, the mixture was directly concentrated and 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; 15-minute gradient, gradient ratio: acetonitrile phase 10% to 30%, flow rate: 25 mL / min) to obtain the title compound 9 (19.4 mg, 20%).

[0211] MS m / z (ESI): 405.1 (M+1) + .

[0212] 1H NMR (400MHz, CD3OD) δ 7.79 - 7.71 (m, 2H), 7.29 (d, 1H), 5.01 (d, 2H), 4.95 (d, 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).

[0213] 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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-1-carboxamide [ka]

[0214] 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]indolizino[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]indolizino[1,2-b]quinoline-11,14-(1H)-dione Compound 6m (700 mg) was separated by high-performance liquid chromatography (GILSON Prep LC with UV detector, chromatography column: Xbridge 5 μm C18 150 x 30 mm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 22% to 32%, flow rate: 50 mL / min) to give compounds 6m-1 (290 mg, short retention time) and 6m-2 (300 mg, long retention time).

[0215] MS m / z (ESI): 450.2 (M+H) +

[0216] 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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-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 completion of the reaction, the mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL × 3). The combined organic phase was washed with saturated brine, dried, and concentrated to give the crude product. The crude product was purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, chromatography column: Xbridge 5 μm C18 150 x 30 mm; mobile phase 1: water (containing 0.1% TFA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 30% to 95%, flow rate: 50 mL / min) to obtain compound 10 (2 mg, yield: 5%).

[0217] MS m / z (ESI): 548.2 (M+H) + .

[0218] 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).

[0219] 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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-2-hydroxyacetamide [ka]

[0220] 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 completion of the reaction, the solvent was rotary evaporated to give the crude product 2-cyclopropyl-2-hydroxyacetic acid 11b (50 mg, white solid), which was used directly in the next step without further purification.

[0221] MS m / z (ESI): 117.1 (M+H) + .

[0222] 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-cycloheptane[3',4':6,7]indolizino[1,2-b]quinolin-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-cycloheptane[3',4':6,7]indolizino[1,2-b]quinolin-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 mixture was stirred at room temperature for 15 minutes. After completion of the reaction, the mixture was analyzed 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; 10-minute gradient, gradient ratio: acetonitrile phase 40% to 50%, flow rate: 20 mL / min). The resulting mixture was purified by the method described above to give compound 11-1 (2.3 mg, yield: 4%) and compound 11-2 (2.5 mg, yield: 4%).

[0223] Single-configuration compound 11-1 (compound with short retention time) MS m / z (ESI): 548.2 (M+H) + .

[0224] 1H NMR (400MHz, 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).

[0225] Single-configuration compound 11-2 (compound with long retention time) MS m / z (ESI): 548.2 (M+H) + .

[0226] 1 H 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).

[0227] 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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-1-carboxamide [ka]

[0228] 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]pyrano[3',4':6,7]indolizino[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]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione Compound 7b (500 mg) was separated by high-performance liquid chromatography (GILSON Prep LC with UV detector, chromatography column: Xbridge 5 μm C18 150 × 30 mm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 22% to 32%, flow rate: 50 mL / min) to give compounds 7b-1 (180 mg, short retention time) and 7b-2 (195 mg, long retention time).

[0229] MS m / z (ESI): 448.2 (M+H) + .

[0230] 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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-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 completion of the reaction, the reaction mixture 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; 10-minute gradient, gradient ratio: acetonitrile phase 45% to 100%, flow rate: 20 mL / min) to obtain compound 12 (3 mg, yield: 11%).

[0231] MS m / z (ESI): 546.2 (M+H) + .

[0232] 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).

[0233] Example 13 (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]indolizino[1,2-b]quinolin-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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0234] Step 1 N-(4-cyclopropyl-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 17a Compound 3f (1.5 g, 5.02 mmol), cyclopropylboronic acid (1.3 g, 15.06 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium chloride (2.2 g, 3.01 mmol), and cesium carbonate (1.6 g, 15.06 mmol) were dissolved in dioxane (15 mL). The mixture was stirred in a microwave reactor at 110 °C for 2 hours. After completion of the reaction, the reaction mixture was poured into water, extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and the combined organic phases were concentrated. Purification by silica gel column chromatography (System B) gave the title compound 17a (1.1 g, yield: 84%).

[0235] MS m / z (ESI): 262.1 (M+1) + .

[0236] Step 2 (Z)-N-(4-cyclopropyl-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 17b Potassium tert-butoxide (0.94 g, 8.4 mmol) was dissolved in a mixture of tetrahydrofuran and tert-butanol (50 mL, V / V = 4:1). Compound 17a (1.1 g, 4.2 mmol) was dissolved in 10 mL of tetrahydrofuran and slowly added to the reaction mixture at 0 °C. After stirring at 0 °C for 10 min, isoamyl nitrite (0.74 g, 6.3 mmol) was added and the reaction mixture was stirred at 0 °C for 50 min. After completion of the reaction, the pH of the mixture was adjusted to 4-5 with dilute hydrochloric acid, extracted three times with ethyl acetate, and the organic phase was washed with saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was rotary evaporated to give crude product 17b, which was used directly in the next step without further purification.

[0237] MS m / z (ESI): 291.0 (M+1) + .

[0238] Step 3 N-(7-amino-4-cyclopropyl-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 17c Compound 17b was added to methanol (50 mL) and palladium-carbon catalyst (500 mg) was added. Under hydrogen gas, the reaction was stirred at room temperature for 2 hours. After the reaction was completed, it was filtered and the filtrate was collected to obtain crude product 17c, which was directly used in the next step without purification.

[0239] MS m / z (ESI): 277.1 (M+1) + .

[0240] Step 4 (9H-Fluoren-9-yl)methyl (8-acetamido-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 17d The pH of the filtrate obtained in the previous step was adjusted to 8-9 with saturated sodium carbonate solution, and then Fmoc-Cl (1.19 g, 4.6 mmol) was added. The reaction was stirred at room temperature for 1 hour. After the reaction was completed, the system was extracted with ethyl acetate, and the organic phase was washed with saturated brine. The organic phase was collected and dried over anhydrous sodium sulfate, and the solvent was evaporated by rotary evaporation to obtain the crude product. Compound 17d (1 g, yield: 48%) was obtained from the crude product using silica gel column chromatography system A.

[0241] MS m / z (ESI): 499.1 (M+1) + .

[0242] Step 5 (9H-Fluoren-9-yl)methyl (8-amino-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 17e Compound 17e (1 g, 2.01 mmol) was dissolved in dioxane (20 mL) and concentrated hydrochloric acid (12 mol / L, 5 mL) was added. The reaction was stirred at 60 °C for 1 hour. After the reaction was completed, the solvent was rotary evaporated to obtain the crude product, which was purified by silica gel column chromatography, System A, to obtain compound 17f (700 mg, yield: 77%).

[0243] MS m / z (ESI): 457.1 (M+1) + .

[0244] 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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-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-pyrano[3,4-f]indolizine-3,6,10(4H)-trione (484 mg, 1.84 mmol) and p-toluenesulfonic acid monohydrate (292 mg, 1.54 mmol) were added. The reaction mixture was stirred at 140 °C for 4 hours. After completion of the reaction, the solvent was rotary evaporated to obtain a crude product. The crude product was purified by silica gel column chromatography, System A, to obtain the title compound 17g (700 mg, yield: 66%).

[0245] MS m / z (ESI): 684.2 (M+1) + .

[0246] Step 7 (9S)-1-Amino-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[d]pyrano[3',4':6,7 indolizino[1,2-b]quinoline-10,13-dione 17h Compound 17g (100 mg, 0.15 mmol) 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 completion of the reaction, the mixture was concentrated in vacuo to give crude compound 17h (60 mg). The crude product was directly used in the next step without further purification.

[0247] MS m / z (ESI): 462.1 (M+1) + .

[0248] 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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide Compound 17h (60 mg, 0.13 mmol) was dissolved in DMF (5 mL), followed by the addition of (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). The reaction was stirred at room temperature for 30 minutes. After completion of the reaction, the reaction mixture 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; 15-minute gradient, gradient ratio: acetonitrile phase 38% to 48%, flow rate: 20 mL / min) to obtain compounds 17-1 (6 mg, yield: 11%) and 17-2 (4 mg, yield: 8%).

[0249] Single-configuration compound 17-1 (short retention time): MS m / z (ESI): 548.2 (M+1) + .

[0250] 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).

[0251] Single stereochemical compound 17-2 (long retention time): MS m / z (ESI): 548.2 (M+1) + .

[0252] 1 H NMR (400MHz, 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).

[0253] Example 14 (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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-1-carboxamide [ka]

[0254] 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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-1-carboxamide Compound 17g (69 mg, 0.15 mmol) 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 completion of the reaction, 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 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 18-min gradient, gradient ratio: acetonitrile phase 35% to 65%, flow rate: 20 mL / min) to give 18-1 (4 mg, yield: 6%) and 18-2 (2.4 mg, yield: 4%).

[0255] Single-configuration compound 18-1 (short retention time): MS m / z (ESI): 560.2 (M+1) + .

[0256] 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).

[0257] Single-configuration compound 18-2 (long retention time): MS m / z (ESI): 560.2 (M+1) + .

[0258] 1 H NMR (400MHz, 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).

[0259] Example 15 (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]indolizino[1,2-b]quinolin-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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide [ka]

[0260] Step 1 5-(5-acetamido-2-bromo-3-fluorophenyl)pent-4-enoic acid 19b Compound 19a (7.0 g, 19.5 mmol) was dissolved in dioxane (60 mL) and water (15 mL). To the solution, pent-4-enoic acid (2.2 g, 22.0 mol), bistriphenylphosphinepalladium dichloride (700 mg, 1.0 mmol), and sodium carbonate (6.2 g, 58.4 mmol) were added, in that order. The reaction mixture was stirred at 90 °C for 16 h under nitrogen gas protection. After completion of the reaction, the mixture was filtered through diatomaceous earth and washed with ethyl acetate. The filtrate was directly concentrated. The crude product was diluted with water (200 mL), washed three times with ether (100 mL x 3), the aqueous phase was adjusted to pH 2 with dilute hydrochloric acid, and extracted with ethyl acetate (100 mL x 3). The organic phases were combined and concentrated to give crude product 19b (6.0 g).

[0261] MS m / z (ESI): 330.1 (M+1) + .

[0262] Step 2 5-(5-acetamido-2-bromo-3-fluorophenyl)pentanoic acid 19c Compound 19b (6.0 g, 18.2 mmol) was dissolved in anhydrous methanol (100 mL) and platinum on carbon (5%, 600 mg) was added. The reaction mixture was purged with hydrogen gas three times and stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was filtered through diatomaceous earth and washed with ethyl acetate. The filtrate was directly concentrated to give crude compound 19c (5.0 g).

[0263] MS m / z (ESI): 332.0 (M+1) + .

[0264] 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 polyphosphoric acid (25 mL). The reaction mixture was stirred at 140 °C for 2 hours. After the reaction was completed, 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 phases were combined and concentrated. The resulting crude product was purified by silica gel column chromatography, System B, to give compound 19d (920 mg, yield: 39%).

[0265] MS m / z (ESI): 314.0 (M+1) + .

[0266] The subsequent synthesis route was the same as in Example 17, except that 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; 15 min gradient, gradient ratio: acetonitrile phase 40% to 95%, flow rate: 30 mL / min) to give products 19-1 (6.5 mg, yield: 9%) and 19-2 (11 mg, yield: 16%).

[0267] Single-configuration compound 19-1 (short retention time): MS m / z (ESI): 562.2 (M+1) + .

[0268] 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).

[0269] Single stereochemical compound 19-2 (long retention time): MS m / z (ESI): 562.2 (M+1) + .

[0270] 1 H NMR (400MHz, 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).

[0271] Example 16 (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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-1-carboxamide [ka]

[0272] 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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-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]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxycyclobutane-1-carboxamide 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 completed, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with saturated brine, dried, and concentrated. The crude product was purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, chromatography column: WELCH Xtimate C18 21.2 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% NH3.HO); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 40% to 100%, flow rate: 30 mL / min) to give compounds 20-1 (2.9 mg, 7%) and 20-2 (1.5 mg, 2%).

[0273] Single-configuration compound 20-1 (short retention time): MS m / z (ESI): 574.7 (M+1) + .

[0274] 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).

[0275] Single-configuration compound 20-2 (long retention time): MS m / z (ESI): 574.7 (M+1) + .

[0276] 1 H NMR (400MHz, 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).

[0277] Example 17 (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-1-yl)-2-hydroxyacetamide [ka]

[0278] 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]indolizino[1,2-b]quinolin-1-yl)carbamate 23a Compound 19j (500 mg, 0.72 mmol) was purified using a Gilson preparative system and a Daicel chiral column (chromatographic column: CHIRALPAK IA 3.0 cm ID × 25 cm, 10 μm; mobile phase 1: MeOH; mobile phase 2: DCM; gradient ratio: MeOH / DCM = 70 / 30, flow rate: 25 mL / min) to separate chiral isomers and give compound 23a (220 mg, yield: 44%).

[0279] 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]indolizino[1,2-b]quinoline-11,14(1H)-dione 23b Compound 23a (45 mg, 0.064 mmol) was dissolved in a mixture of N,N-dimethylformamide and ethylenediamine (7 mL, V / V = 5:2), and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was directly concentrated to give crude compound 23b (30 mg), which was used directly in the next step without further purification.

[0280] MS m / z (ESI): 476.2 (M+1) + .

[0281] Step 3 (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-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), benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (22 mg, 0.051 mmol), and N,N-diisopropylethylamine (11 mg, 0.084 mmol) were added. The reaction was stirred at room temperature for 3 hours. After completion of the reaction, 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 x 250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 46% to 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%).

[0282] 33-1 (single configuration compound, short retention time) MS m / z (ESI): 574.2 (M+1) + .

[0283] 1H NMR (400MHz, 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).

[0284] 33-2 (Single stereochemical compound, long retention time) MS m / z (ESI): 574.2 (M+1) + .

[0285] 1 H NMR (400MHz, 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), 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).

[0286] Example 18 (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide [ka]

[0287] 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]indolizino[1,2-b]quinolin-1-yl)carbamate 21a Compound 17f (600 mg, 1.02 mmol) was purified using a Gilson preparative system and a Daicel chiral column (chromatographic column: CHIRALPAK IB 3.0 cm ID × 25 cm, 10 μm; mobile phase 1: MeOH; mobile phase 2: DCM; gradient ratio: MeOH / DCM = 90 / 10, flow rate: 25 mL / min) to separate chiral isomers and give compound 21a (250 mg, yield: 42%).

[0288] 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]pyrano[3',4':6,7]indolizino[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 completion of the reaction, the solvent was evaporated in vacuo to give the title compound 21b (34 mg). The product was used directly in the next step without purification.

[0289] MS m / z (ESI): 462.1 (M+1) + .

[0290] Step 3 (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-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 completion of the reaction, 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; 15-minute gradient, gradient ratio: acetonitrile phase 40% to 100%, flow rate: 30 mL / min) to obtain products 34-1 (4.2 mg, yield: 11%) and 34-2 (3.9 mg, yield: 10%).

[0291] 34-1 (single configuration compound, compound with short retention time) MS m / z (ESI): 574.2 (M+1) + .

[0292] 1 H NMR (400MHz, 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).

[0293] 34-2 (single configuration compound, compound with long retention time) MS m / z (ESI): 574.2 (M+1) + .

[0294] 1H NMR (400MHz, 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).

[0295] Example 19 (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypentanamide [ka]

[0296] 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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-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 mixture was allowed to react at room temperature for 1 hour. After completion of the reaction, the mixture was purified by preparative high-performance liquid chromatography (GILSON Prep LC with UV detector, chromatography column: Xbridge 10 μm C18 250 x 30 mm, 10 μm; mobile phase 1: water (containing 10 mmol / L FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile 5% to 57%, flow rate: 50 mL / min) to give compounds 36-1 (6.98 mg, yield: 18%) and 36-2 (6.70 mg, yield: 18%).

[0297] 36-1 (single configuration compound, compound with short retention time) MS m / z (ESI): 550.2 (M+1) + .

[0298] 1H 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).

[0299] 36-2 (single configuration compound, compound with long retention time) MS m / z (ESI): 550.2 (M+1) + .

[0300] 1 H NMR (400MHz, 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).

[0301] Example 20 (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxy-4-methylpentanamide [ka]

[0302] Step 1 (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-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 mixture was incubated at room temperature for 1 hour. After completion of the reaction, the mixture was analyzed 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; 15 min gradient, gradient ratio: acetonitrile phase 42% to 100%, flow rate: 20 mL / min). The resulting mixture was purified by HPLC to give compounds 37-1 (6.6 mg, yield: 18%) and 37-2 (6.9 mg, yield: 18%).

[0303] 37-1 (single configuration compound, compound with short retention time) MS m / z (ESI): 564.2 (M+1) + .

[0304] 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).

[0305] 37-2 (single configuration compound, compound with long retention time) MS m / z (ESI): 564.2 (M+1) + .

[0306] 1 H NMR (400MHz, 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).

[0307] Example 21 (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide [ka]

[0308] (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]indolizino[1,2-b]quinolin-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]indolizino[1,2-b]quinolin-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 mixture was allowed to react at room temperature for 1 hour. After completion of the reaction, the product was purified and separated by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, chromatography column: Xbridge 5 μm C18 150 × 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 45% to 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%).

[0309] 38-1 (single configuration compound, compound with short retention time) MS m / z (ESI): 562.3 (M+1) + .

[0310] 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).

[0311] 38-2 (single configuration compound, compound with long retention time) MS m / z (ESI): 562.3 (M+1) + .

[0312] 1 H NMR (400MHz, 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).

[0313] Example 22 (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]indolizino[1,2-b]quinolin-1-yl)-3-hydroxy-2-methylbutanamide 39 [ka]

[0314] 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]indolizino[1,2-b]quinolin-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-methylbutyric acid (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 completion of the reaction, the residue 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, 5 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 10-minute gradient, gradient ratio: acetonitrile phase 42% to 52%, flow rate: 25 mL / min) to obtain compound 39 (2 mg, yield: 5%).

[0315] MS m / z (ESI): 550.0 (M+1) + .

[0316] 1H NMR (400MHz, 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).

[0317] Biological evaluation The present invention will now be further described with reference to test examples, but these test examples are not intended to limit the scope of the present invention.

[0318] Test Example 1: Growth inhibition experiment of SK-BR-3 cells by compounds 1.1 Experimental materials [Table 1]

[0319] 1.2 Experimental equipment [Table 2]

[0320] 1.3 Test Method (1) Cell plating: First, tumor cells SK-BR-3 were cultured in the corresponding medium, digested with trypsin, centrifuged, resuspended, and counted. The cells were then adjusted to the appropriate concentration and plated onto a 384-well plate.

[0321] (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 ECHO, and the final concentration of DMSO in the wells of the cell culture plate was 0.33%, and the cells were incubated in an incubator at 37°C and 5% CO2 for 72 h.

[0322] (3) After incubation, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the wells were shaken for 2 minutes in a high-speed shaker, centrifuged at 1000 rpm for 1 minute, and then placed in the dark at room temperature for 30 minutes. The chemiluminescence signal was then read using an Envision instrument.

[0323] (4) Detection of cell activity: IC using GraphPad Prism 8 software 50 Calculate the IC of the compound using the following nonlinear fitting equation: 50 was obtained (see Table 1).

[0324] Y=Bottom + (Top-Bottom) / (1+10^((LogIC 50 -X)* HillSlope)) Y: inhibition rate, X: compound concentration log value, Inhibition % (%) = 100 - (compound well reading - low read control well reading) / (high read control well reading - low read control well reading) * 100; High readout control wells: Add 100 nL of DMSO to the cells. Low reading control wells: wells without cells.

[0325] [Table 3]

[0326] As the results show, the compounds of the present invention have a strong growth inhibitory effect on the proliferation of SK-BR-3 cells.

[0327] Test Example 2: Growth inhibition experiment of MDA-MB-468 cells by test compounds 2.1 Experimental materials [Table 4]

[0328] 2.2 Experimental equipment [Table 5]

[0329] 2.3 Test Method (1) Cell plating: First, MDA-MB-468 tumor cells were cultured in the corresponding medium, digested with trypsin, centrifuged, resuspended, and counted. The cells were then adjusted to the appropriate concentration and plated onto a 384-well plate.

[0330] (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 ECHO, and the final concentration of DMSO in the wells of the cell culture plate was 0.33%, and the cells were incubated in an incubator at 37°C and 0% CO2 for 72 h.

[0331] (3) After incubation, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the wells were shaken for 2 minutes in a high-speed shaker, centrifuged at 1000 rpm for 1 minute, and then placed in the dark at room temperature for 30 minutes. The chemiluminescence signal was then read using an Envision instrument.

[0332] (4) Detection of cell activity: IC using GraphPad Prism 8 software 50 Calculate the IC of the compound using the following nonlinear fitting equation: 50 was obtained (see Table 2).

[0333] Y=Bottom + (Top-Bottom) / (1+10^((LogIC 50-X)* HillSlope)) Y: inhibition rate, X: compound concentration log value, Inhibition % (%) = 100 - (compound well reading - low read control well reading) / (high read control well reading - low read control well reading) * 100; High readout control wells: Add 100 nL of DMSO to cells Low reading control wells: wells without cells.

[0334] [Table 6]

[0335] As the results show, the test compound has a strong growth inhibitory effect on the proliferation of MDA-MB-468 cells.

[0336] Test Example 3: Growth inhibition experiment of NCI-N87 cells by test compounds 3.1 Experimental materials [Table 7]

[0337] 3.2 Experimental equipment [Table 8]

[0338] 3.3 Test Method (1) Cell plating: First, NCI-N87 tumor cells were cultured in the corresponding medium, digested with trypsin, centrifuged, resuspended, and counted. The cells were then adjusted to the appropriate concentration and plated onto a 384-well plate.

[0339] (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 ECHO, and the final concentration of DMSO in the wells of the cell culture plate was 0.33%, and the cells were incubated in an incubator at 37°C and 0% CO2 for 72 h.

[0340] (3) After incubation, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the wells were shaken for 2 minutes in a high-speed shaker, centrifuged at 1000 rpm for 1 minute, and then placed in the dark at room temperature for 30 minutes. The chemiluminescence signal was then read using an Envision instrument.

[0341] (4) Detection of cell activity: IC using GraphPad Prism 8 software 50 Calculate the IC of the compound using the following nonlinear fitting equation: 50 was obtained (see Table 3).

[0342] Y=Bottom + (Top-Bottom) / (1+10^((LogIC 50 -X)* HillSlope)) Y: inhibition rate, X: compound concentration log value, Inhibition % (%) = 100 - (compound well reading - low read control well reading) / (high read control well reading - low read control well reading) * 100; High readout control wells: Add 100 nL of DMSO to the cells. Low reading control wells: wells without cells.

[0343] [Table 9]

[0344] As the results show, the test compound has a strong growth inhibitory effect on the proliferation of NCI-N87 cells.

[0345] Test Example 4 Dynamic solubility test in PBS 7.4 buffer solution 4.1 Experimental steps 1) Preparation of stock solutions 10 mM stock solutions of the measurement targets 6-2, 11-2, 17-2, Dxd, and the control drugs progesterone and diclofenac were prepared using DMSO.

[0346] 2) Dynamic solubility measurement step 15 μL of 10 mM stock solution was taken and added to the corresponding locations of a 96-well plate in the specified order. 485 μL of PBS 7.4 buffer was added to the corresponding vials in the sample plate. The experiment was performed in duplicate. A stir bar was placed in each vial and the vial was capped. The sample tray was then placed in a thermomixer and shaken at 1100 rpm at 25°C for 2 hours. After 2 hours, the lid was removed, the stir bar was sucked in with a large magnet, and the sample was then transferred from the sample plate to a filter plate. Negative pressure was generated with a vacuum pump, and the sample was filtered. 5 μL of the filtrate and 5 μL of blank DMSO were transferred to a new sample plate, followed by the addition of 490 μL of internal standard water (acetonitrile:water = 1:1) containing the internal standard. Depending on the peak shape, the sample dilution may be diluted with a specific ratio of internal standard water to obtain a better peak.

[0347] 3) Preparation of 3 μM standard solution Six microliters of the 10 mM DMSO stock solution was transferred to an empty plate, and 194 μL of DMSO was added to make a 300 μM standard solution. Five microliters of the 300 μM standard solution was transferred to another empty plate, and 5 μL of blank buffer and 490 μL of internal standard water (acetonitrile:water = 1:1) containing the internal standard were added to make the final standard concentration 3 μM.

[0348] 4) Sample analysis step The sample plate was placed in the sample tray of an autosampler and the samples were evaluated by liquid chromatography mass spectrometry.

[0349] 4.0 Data Analysis All calculations were performed in Microsoft Excel.

[0350] The analysis and quantification of the sample filtrate was performed by qualitative and quantitative analysis of standard peaks of known concentrations using chromatography-mass spectrometry. The calculation formulas for the solubility values ​​of the control drug and the measured substance were as follows:

number

[0351] 4.2 Experimental results The experimental results are shown in Table 4.

[0352] [Table 10]

[0353] As the results show, the solubility of the representative compounds 6-2 and 11-2 of the present application in PBS 7.4 buffer solution is clearly superior to that of Dxd.

[0354] Test Example 5 Pharmacokinetics test in SD rats After a single intravenous injection (2mpk) of compounds 6-2, 11-2, 17-2 and Dxd into male SD rats (8 blood sampling times: 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 8h and 24h after IV administration), the mean pharmacokinetic parameters in plasma were as shown in Table 5. [Table 11] As the results show, the representative compounds of the present application are rapidly eliminated in the body of rats, have good safety profiles, and have good pharmacokinetic properties.

[0355] Test Example 6: Inhibition of human liver microsomal CYP1A2, CYP2D6, and CYP3A4 by compounds 6.1 Subtype Positive Control Inhibitors [Table 12]

[0356] 6.2 Preparation of substrate stock solution [Table 13]

[0357] 6.3 Experimental Process Incubations were performed in a 96-deep-well plate. Each well of the incubation plate was added with 169 μL of the "main solution" and 1 μL of multiple concentrations of test compound or positive control compound (DMSO). The incubation plate was preincubated in a water bath at 37°C for 5 minutes. Next, 10 μL of diluted substrate solution was added to the incubation plate, and the plate was incubated for 15 seconds while mixing on a vortex mixer. The reaction was then initiated by adding 20 μL of 10 mM NADPH solution to a final concentration of 1 mM. At predetermined time points, the reaction was quenched by adding 300 μL of stop solution (cold acetonitrile + 3% formic acid, 200 nM alprazolam, 200 nM labetalol hydrochloride, 200 nM tolbutamide). The plate was centrifuged at 3220 g for 40 minutes. 150 μL of the supernatant was transferred to a new plate. The supernatant may also be diluted with 150 μL of pure water. The mixture was mixed uniformly and the content of substrate metabolites was measured by LC / MS / MS.

[0358] 6.4 Data Analysis An automated peak integration area check was performed for all samples. Analytical and internal standard peak areas were exported to an excel sheet. Inhibition of each P450 enzyme in human liver microsomes was measured by the percent reduction in marker metabolite formation activity compared to the uninhibited control (= 100% activity). IC was calculated as the residual activity (%) and the logarithm of the inhibitor concentration. 50 The value of was calculated.

[0359] The percent remaining activity was calculated as follows: Area ratio = analyte peak area / internal standard peak area Residual activity (%)= 測定対象薬物の面積比 / ブランク対照の面積比 *100% IC using Excel XLfit 5.5.1.3 50 The value of was calculated.

[0360] The experimental results are shown in Table 6. [Table 14]

[0361] As the results show, the representative compounds of the present application have low inhibitory activity against CYP enzymes and good safety.

[0362] Test Example 7: Stability test of compound in human plasma 7.1 Experimental process (1) 199 μL of human plasma was added to each cell culture plate, and the culture plate was preheated to 37° C. and kept incubate for 15 minutes.

[0363] (2) After preincubation, 1 μL of a 1 mmol / L test compound and 1 μL of a 1 mmol / L control compound were added. 199 μL of plasma was added to achieve a final concentration of 5 μmol / L of the test compound and 5 μmol / L of the control compound. The final concentration of the organic solvent was 0.5%. The experiment was repeated twice.

[0364] (3) The reaction samples were incubated at 37°C.

[0365] (4) The reaction was stopped at 0, 1, 2, 6, and 24 hours by adding 600 μL of cold methanol containing the internal standard. All samples were vortex mixed for 10 minutes and then centrifuged at 3220 g for 30 minutes to precipitate the proteins. 100 μL of the supernatant was transferred to a new plate. The supernatant was diluted with ultrapure water according to the signal response and peak shape of liquid chromatography-mass spectrometry (LC-MS).

[0366] 7.2 Data analysis All calculations were performed using Microsoft Excel. Peak area ratios were determined from extracted ion chromatograms. The percentage of compound remaining at each time point was calculated by the following formula: Remaining Percentage tmin (%) = peak area ratio tmin / peak area t0 × 100 The experimental results are shown in Table 7.

[0367] [Table 15]

[0368] As the results show, the representative compounds of the present invention, 6-2, 11-2, and 17-2, have good stability in human plasma.

[0369] The above are examples of the embodiments of the technical solutions of the present invention. It should be understood that the protection scope of the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made by those skilled in the art within the spirit and principle of the present invention should be included in the protection scope of the claims of this application.

Claims

1. A compound of formula I, its racemate, stereoisomer, tautomer, isotopically labeled form, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, 【Chemistry 1】 Here, R １ , R ２ , R ３ are the same or different and independently represent 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 groups, R ４ is H or 【Chemistry 2】 and R ４１ is 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 group, (C １～６ alkyl) ２ N-C １～６ Alkyl group, C １～６ Alkoxyalkyl group, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3- to 6-membered heterocyclyl group, C ６～１４ selected from aryl groups and 5- to 14-membered heteroaryl groups; R ５ is H, 【Transformation 3】 and R ５１ , R ５２ are the same or different, and independently represent 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 group, (C １～６ alkyl) ２ N-C １～６ Alkyl group, C １～６ Alkoxyalkyl group, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3- to 6-membered heterocyclyl group, C ６～１４ aryl groups and 5- to 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 group, (C １～６ alkyl) ２ N-C １～６ Alkyl group, C １～６ Alkoxyalkyl group, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group, 3- to 6-membered heterocyclyl group, C ６～１４ aryl groups and 5- to 14-membered heteroaryl groups, and ring A is C ３～８ Cycloalkyl group or C ３～８ heterocyclyl groups, and Ra is H, a hydroxy group, CN, a halogen, C １～６ Alkyl group, C １～６ 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; m is selected from integers of 0 to 6.

2. R １ H, OH, CN, halogen, C １～６ Alkyl group, C ２～６ Alkenyl group, C ２～６ Alkynyl group, C ３～６ Cycloalkyl group or halo C １～６ alkoxy groups, Preferably, R １ H, OH, CN, halogen, C １～６ Alkyl group, C ２～６ Alkenyl group, C ２～６ Alkynyl group or halo C １～６ alkoxy groups, Preferably, R １ is selected from H, OH, Br, methyl, difluoromethoxy, 2,2,2-trifluoroethoxy, vinyl, cyclopropyl, or ethynyl; Preferably, R １ is selected from H, OH, Br, a methyl group, a difluoromethoxy group, a 2,2,2-trifluoroethoxy group, a vinyl group, or an ethynyl group. Preferably, R ２ is H, halogen, CN or C １～６ alkyl groups, Preferably, R ２ is selected from H or F; Preferably, R ３ is H or C １～６ alkyl groups, Preferably, R ３ The compound of 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】 is selected from Preferably, X-R ４ teeth 【Transformation 7】 and preferably 【Transformation 8】 and Preferably, X-R ４ Ha-CH ２ 3. The compound according to claim 1 or 2, characterized in that:

4. R ５ is H, 【Chemistry 9】 and R ５１ is selected from H, methyl, ethyl, isopropyl, or cyclopropyl; R ５２ is selected from H or a methyl group, R ５３ is selected from a methyl group, and the A ring is C ３～６ cycloalkyl groups, and Ra is H, a hydroxy group, CN, a halogen, or C １～６ Alkyl group, C １～６ 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, R ５ is H, 【Chemistry 10】 is selected from Preferably, R ５ is H, 【Chemistry 11】 is selected from Preferably, R ５１ is selected from H, methyl, ethyl, isopropyl, or cyclopropyl; R ５２ is selected from H or a methyl group, R ５３ is selected from a methyl group, and ring A is selected from a cyclobutane ring; Preferably, R ５ is H, 【Chemistry 12】 Compounds according to any one of claims 1 to 3, characterized in that they are selected from:

5. X is selected from CH or N, and when X is CH, R ４ is H, or when X is N, R ５ is H, Preferably, compounds according to any one of claims 1 to 4, characterized in that m is selected from 0, 1 or 2.

6. The structure of the compound of formula I is shown below: 【Chemistry 13】 Here, R １ , R ２ , R ４ , R ５ , X, m independently of one another have the definitions according to any one of claims 1 to 5, Preferably, the structure of the compound of formula I is as shown below: 【Chemistry 14】 Here, R １ , R ２ , R ５ have, independently of one another, the definitions according to any one of claims 1 to 5, Preferably, the structure of the compound of formula I is as shown below: 【Chemistry 15】 Here, R １ , R ２ , R ５１ , R ５２ 6. The compound according to claim 1, wherein ring A, Ra, m, n and q independently have the definitions according to claim 1.

7. The structure of the compound of formula I is: 【Chemistry 16】 【change】 【change】 【change】 【change】 【change】 The compound according to any one of claims 1 to 6, characterized in that it is as shown in

8. A structural fragment D having a structure after dehydrogenation of a compound of formula I according to any one of claims 1 to 7, Preferably, the structure of D is: 【Chemistry 17】 【change】 【change】 【change】 【change】 【change】 Structural fragment D, which is as shown in

9. A compound of formula V, its racemate, stereoisomer, tautomer, isotopically labeled form, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, L'-D (Formula V) wherein L' is a linker containing a linker moiety M capable of reacting with an antibody or antigen-binding fragment thereof, and L'-D reacts with the antibody or antigen-binding fragment thereof, after which L' forms a linker L; Preferably, L' is a peptide residue L １ and a fragment L in which the peptide residue is bound to D ２ Including, A compound wherein D has the definition set forth in claim 8.

10. 1. An antibody drug conjugate as shown in Formula VI, A-[L-D] β (Form VI) An antibody-drug conjugate, wherein Ab is an antibody or an antigen-binding fragment thereof, D has the definition of claim 8, L is a linker connecting Ab and D, and β is selected from integers or decimals between 1 and 10.

11. A method for producing the compound according to any one of claims 1 to 7, comprising the following method 1 or method 2: Method 1 is Compound I-41 to the protecting group PG ４ to obtain compound I-42; and step (2) reacting compound I-42 with compound I-43 to obtain a compound of formula I, [Chemistry 18] Here, R １ , R ２ , R ３ , R ４ , R ５ , X, m have the definitions according to any one of claims 1 to 7, Y is a leaving group selected from, for example, OH, Cl, Br, I, PG ４ is selected from amino protecting groups, e.g., Fmoc, Boc, Bn, Cbz; Method 2 is Compound I-51 to the protecting group PG ５ to obtain compound I-52; and (2) reacting compound I-52 with compound I-53 to obtain a compound of formula I, 【Chemistry 19】 Here, R １ , R ２ , R ３ , R ４ , R ５ , R ５１ , X, m, n have the definitions according to any one of claims 1 to 7, Y is a leaving group selected from, for example, OH, Cl, Br, I, PG ５ is an amino protecting group, for example selected from Fmoc, Boc, Bn, Cbz.

12. A therapeutically effective amount of at least one of the compounds of any one of claims 1 to 7, its racemate, stereoisomer, tautomer, isotopically labeled, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound thereof, Preferably, a pharmaceutical composition comprising a therapeutically effective amount of the antibody-drug conjugate of claim 10.

13. Use of at least one of the compound according to any one of claims 1 to 7, the compound represented by formula V according to claim 9, or the antibody-drug conjugate represented by formula VI according to claim 10, or a racemate, stereoisomer, tautomer, isotope-labeled form, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound thereof, or the pharmaceutical composition according to claim 12, in the manufacture of a topoisomerase I inhibitor and / or in the manufacture of a medicament for preventing or treating a disease or disorder associated with topoisomerase I, Preferably, the disease or disorder is a tumor, and the tumor comprises 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.

14. A method for treating a neoplastic disease, comprising administering to a patient a prophylactically or therapeutically effective amount of at least one of a compound of Formula I or Formula V or an antibody-drug conjugate of Formula VI, a racemate, a stereoisomer, a tautomer, an isotopically labeled compound, a solvate, a polymorph, a pharmaceutically acceptable salt, or a prodrug compound thereof, Preferably, the tumor comprises 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.

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