Camptothecin derivative, pharmaceutical composition as well as preparation method and application of camptothecin derivative

Abstract

The invention relates to a camptothecin derivative as shown in a general formula I, a pharmaceutical composition as well as a preparation method and application of the camptothecin derivative. The camptothecin derivative has good tumor inhibition activity, and can be used for treating or preventing tumors and preparing medicines for treating or preventing tumor diseases. # imgabs0 #

Description

Camptothecin derivatives, pharmaceutical compositions, preparation methods and applications thereof

[0001] This application claims priority to six applications with application numbers 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, all with the invention name “Camptothecin derivatives, pharmaceutical compositions, preparation methods and applications thereof”. Technical Field

[0002] The present invention belongs to the field of medicine, and in particular relates to a camptothecin derivative, a pharmaceutical composition, and a preparation method and application thereof. Background Art

[0003] Camptothecin (CPT) is a natural product isolated from Camptotheca acuminata, a plant of the Davidiaceae family. It is a pentacyclic compound composed of a quinoline ring (AB), a pyrrole ring (C), a pyridone ring (D), and an α-hydroxylactone ring (E), with the 20-position in the S configuration (see the structural formula below). Due to its excellent anticancer activity, it was introduced to clinical practice in the early 1970s. However, clinical trials were terminated due to the development of severe side effects such as diarrhea and hemorrhagic cystitis.

[0004] Research data show that camptothecin can form a ternary complex with DNA topoisomerase I in cells, thereby inhibiting DNA unwinding, leading to DNA replication blockage, and thus causing cell death (Cancer Res. 1989, 49, 6365). Camptothecin and its derivatives have strong anti-tumor activity in animal models such as lung cancer, breast cancer, colorectal cancer, and ovarian cancer (Nature Review Cancer. 2006, 6, 789). Currently, multiple camptothecin drugs have been approved for marketing for tumor treatment (Med Res. Rev. 2015, 35, 753). Irinotecan is a drug for the treatment of colorectal cancer; topotecan is used for the treatment of ovarian cancer; belotecan is used for the treatment of ovarian cancer and small cell lung cancer. Camptothecin derivatives also include ixitecan, rubitecan, diflutecan, lortotecan, gimatecan, ximintecan, gimitecan, ilinotecan, etc. Camptothecin drugs or their derivatives often have hematotoxicity caused by bone marrow suppression, such as leukopenia, thrombocytopenia, anemia, neutropenia, etc., as well as gastrointestinal side effects such as nausea, vomiting, and diarrhea. Clinical studies have found that measures to improve the safety and efficacy of camptothecin compounds include increasing water solubility, improving their pharmacokinetic properties, increasing activity, reducing dosage, or using their conjugates with antibodies to form antibody-drug conjugates. Therefore, the development of camptothecin compounds and their conjugates with novel structures that can improve efficacy and ameliorate safety issues still has high clinical demand and application value.

[0005] Summary of the Invention

[0006] The present invention provides a compound represented by Formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound:

[0007] Wherein, R1, R2, R3 are the same or different and are independently selected from H, OH, CN, halogen, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 1-10 Alkoxy, halogenated C 1-10 Alkyl, halogenated C 1-10 Alkoxy, cyano C 1-10 Alkyl, cyano C 1-10 Alkoxy, C 3-10 Cycloalkyl;

[0008] R4 is selected from H or R 41 Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl-NH-, (C 1-6Alkyl)2N-, C 1-6 Alkyl-NH-C 1-6 Alkyl, (C 1-6 alkyl)2N-C 1-6 Alkyl, C 1-6 Alkoxyalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-14 Aryl and 5-14 membered heteroaryl;

[0009] R5 is selected from H, R 51 、R 52 The same or different, independently selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl-NH-, (C 1-6 Alkyl)2N-, C 1-6 Alkyl-NH-C 1-6 Alkyl, (C 1-6 alkyl)2N-C 1-6 Alkyl, C 1-6 Alkoxyalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-14 Aryl and 5-14 membered heteroaryl; R 53 Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl-NH-, (C 1-6 Alkyl)2N-, C 1-6 Alkyl-NH-C 1-6 Alkyl, (C 1-6 alkyl)2N-C 1-6 Alkyl, C 1-6 Alkoxyalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-14 Aryl and 5-14 membered heteroaryl; Ring A is selected from C 3-8 Cycloalkyl or 3-8 membered heterocyclic group, Ra is selected from H, hydroxy, CN, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl; n is selected from 0, 1 or 2; q is selected from 0, 1 or 2;

[0010] X is selected from CH or N;

[0011] m is an integer selected from 0-6.

[0012] According to some embodiments, R1 is selected from H, OH, CN, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl or halogenated C 1-6 alkoxy;

[0013] According to some embodiments, R1 is selected from H, OH, CN, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl or halo C 1-6 alkoxy;

[0014] According to some embodiments, R1 is selected from H, OH, Br, methyl, difluoromethoxy, 2,2,2-trifluoroethoxy, vinyl, cyclopropyl or ethynyl;

[0015] According to some embodiments, R1 is selected from H, OH, Br, methyl, difluoromethoxy, 2,2,2-trifluoroethoxy, vinyl, or ethynyl.

[0016] According to some embodiments, R2 is selected from H, halogen, CN or C 1-6 alkyl;

[0017] According to some embodiments, R2 is selected from H or F.

[0018] According to some embodiments, R3 is selected from H or C 1-6 alkyl;

[0019] According to some embodiments, R3 is H.

[0020] According to some embodiments, R4 is selected from H or For example

[0021] According to some embodiments, R4 is selected from H or

[0022] According to some embodiments, X-R4 is Preferably

[0023] According to some embodiments, X-R4 is -CH2-.

[0024] According to some embodiments, R5 is selected from H, Among them, R 51 is selected from H, methyl, ethyl, isopropyl or cyclopropyl; R 52 is selected from H or methyl; R 53 is selected from methyl; Ring A is selected from C 3-6Cycloalkyl; Ra is selected from H, hydroxy, CN, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl; n is selected from 0 or 1; q is selected from 0 or 1;

[0025] According to some embodiments, ring A is selected from a cyclobutane ring.

[0026] According to some embodiments, R5 is selected from H,

[0027] According to some embodiments, R 51 is selected from H, methyl, ethyl, isopropyl or cyclopropyl; R 52 is selected from H or methyl; R 53 is selected from methyl; Ring A is selected from cyclobutane ring;

[0028] According to some embodiments, R5 is selected from H,

[0029] According to some embodiments, R5 is selected from H,

[0030] 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;

[0031] According to some embodiments, m is selected from 0, 1 or 2.

[0032] According to some embodiments, the compound structure shown in Formula I is as follows:

[0033] wherein R1, R2, R4, R5, X, and m are independently defined as described herein.

[0034] According to some embodiments, the compound structure shown in Formula I is as follows:

[0035] wherein R1, R2, and R5 are independently defined as described herein.

[0036] According to some embodiments, the compound structure shown in Formula I is as follows:

[0037] Among them, R1, R2, R 51 、R 52 , Ring A, Ra, m, n, q are independently defined as described herein.

[0038] According to an embodiment of the present invention, the structure of the compound of formula I is shown below:

[0039] According to some embodiments, the present invention also provides a structural fragment D of the compound of Formula I described herein after dehydrogenation;

[0040] According to some embodiments, the structure of D is as follows:

[0041] The present invention also provides a compound represented by Formula V, its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound: L'-D (Formula V)

[0042] wherein L' is a linker containing a linker portion M capable of reacting with an antibody or an antigen-binding fragment thereof, and after L'-D reacts with the antibody or an antigen-binding fragment thereof, L' forms a linker L;

[0043] Preferably, L' comprises the peptide residue L1 and a fragment L2 where the peptide residue is linked to D.

[0044] The definition of D is as shown in this article.

[0045] The present invention also provides an antibody drug conjugate of formula VI, Ab-[LD] β (Formula VI)

[0046] Wherein, Ab is an antibody or an antigen-binding fragment thereof, D has the definition as described in the text of the application, L is a linker connecting Ab and D, and β is selected from an integer or decimal between 1-10.

[0047] The present invention also provides a method for preparing the compound of formula I, comprising the following scheme 1 or scheme 2:

[0048] Option 1: includes the following steps:

[0049] (1) Compound I-41 is deprotected by PG4 to obtain compound I-42;

[0050] (2) Compound I-42 reacts with compound I-43 to obtain the compound of formula I;

[0051] wherein R1, R2, R3, R4, R5, X, and m have the definitions described herein; Y is selected from a leaving group, such as OH, Cl, Br, and I; and PG4 is selected from an amino protecting group, such as Fmoc, Boc, Bn, and Cbz.

[0052] Option 2: includes the following steps:

[0053] (1) Compound I-51 is deprotected by PG5 to obtain compound I-52;

[0054] (2) Compound I-52 reacts with compound I-53 to obtain the compound of formula I;

[0055] Among them, R1, R2, R3, R4, R5, R 51 , X, m, n have the definitions described herein; Y is selected from a leaving group, such as OH, Cl, Br, I; PG5 is selected from an amino protecting group, such as Fmoc, Boc, Bn, Cbz.

[0056] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds represented by Formula I or Formula V, their racemates, stereoisomers, tautomers, isotope-labeled substances, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof.

[0057] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the antibody-drug conjugate represented by Formula VI.

[0058] According to an embodiment of the present invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0059] According to an embodiment of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

[0060] The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventively or therapeutically effective amount of a compound represented by Formula I or Formula V, or an antibody-drug conjugate represented by Formula VI, or at least one of their racemates, stereoisomers, tautomers, isotope-labeled forms, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds.

[0061] The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventive or therapeutically effective amount of the above-mentioned pharmaceutical composition.

[0062] The tumor disease is selected from breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal cancer, melanoma or leukemia.

[0063] In some embodiments, the patient comprises a mammal, preferably a human.

[0064] The present invention also provides at least one of the compounds represented by Formula I, Formula V, or the antibody-drug conjugate represented by Formula VI, their racemates, stereoisomers, tautomers, isotope-labeled substances, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds for treating tumor diseases, or pharmaceutical compositions thereof.

[0065] The present invention also provides the use of at least one of the compounds represented by Formula I, Formula V, or the antibody-drug conjugate represented by Formula VI, their racemates, stereoisomers, tautomers, isotope-labeled products, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof, or the pharmaceutical compositions thereof, in the preparation of topoisomerase I inhibitors and / or in the preparation of drugs for preventing or treating diseases or conditions associated with topoisomerase I.

[0066] In some embodiments, the disease or condition is a tumor, which includes breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal cancer, melanoma or leukemia. Beneficial effects

[0067] The compounds provided by the present invention have good tumor inhibitory effects and can be used to treat or prevent cancer (such as breast cancer or gastric cancer), as well as to prepare drugs for treating or preventing such conditions and diseases.

[0068] Definitions and Explanations of Terms

[0069] Unless otherwise indicated, the definitions of groups and terms in this specification and claims, including definitions used as examples, exemplary definitions, preferred definitions, definitions in tables, and definitions of specific compounds in the Examples, may be arbitrarily combined and coupled with one another. The group definitions and compound structures resulting from such combinations and couplings should be understood to be within the scope of this specification and / or claims.

[0070] Unless otherwise indicated, numerical ranges recited in this specification and claims are equivalent to reciting at least each specific integer value therein. For example, the numerical range "1-12" is equivalent to reciting each integer value in the numerical range "1-12," namely, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12. In addition, when a numerical range is defined as a "number," it should be understood that both endpoints of the range, each integer within the range, and each decimal within the range are recited.

[0071] The term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0072] “C 1-10 "Alkyl" means straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, "C 1-8 "Alkyl" means straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms, "C 1-6 The term "alkyl" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl group is, for example, 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 the like or isomers thereof.

[0073] “C 2-10 "Alkenyl" is understood to mean preferably a linear or branched monovalent hydrocarbon radical 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 Alkenyl". "C 2-10 "Alkenyl" is understood to mean preferably a linear or branched monovalent hydrocarbon radical containing one or more double bonds and having 2, 3, 4, 5, 6, 7 or 8 carbon atoms, e.g. 2-6 alkenyl), having 2 or 3 carbon atoms (i.e., C 2-3It is understood that when the alkenyl group contains more than one double bond, the double bonds may be separated from one another or conjugated. The alkenyl group is, for example, vinyl, allyl, (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-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2-enyl, (Z)-1-methyl But-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-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.

[0074] The term "C 2-10 "Alkynyl" is understood to mean preferably a linear or branched monovalent hydrocarbon radical containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, e.g. having 2, 3, 4, 5, 6, 7 or 8 carbon atoms (i.e. "C 2-8 alkynyl”), having 2, 3, 4, 5, or 6 carbon atoms (i.e., “C 2-6 Alkynyl”), having 2 or 3 carbon atoms (“C 2-3The alkynyl group is, for example, ethynyl, 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-methylpent-4-ynyl, 1-methylpent-4-ynyl, In some embodiments, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.

[0075] The term "C 3-10 "Cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic (eg bridged, spiro) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. 3-10 The 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 borneol, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, 2,6-diazaspiro[3,4]octyl, or a tricyclic hydrocarbon group such as adamantyl.

[0076] Unless otherwise defined, the term "3-6 membered heterocyclyl" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5- or 6-membered monocyclic ring, and containing at least one, for example 1, 2, 3, 4, 5 or more heteroatoms selected from O, S and N, wherein N and S may also be optionally oxidized to various oxidation states to form nitrogen oxides, -S(O)- or -S(O)2- states. The heterocyclyl may include fused or bridged rings and spirocyclic rings. In particular, the heterocyclyl may include, but is not limited to, a 4-membered ring such as azetidinyl, oxetanyl; a 5-membered ring such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl.

[0077] The term "C 6-14 "Aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring ("C 6-14 or a ring having 9 carbon atoms ("C9 aryl"), for example indanyl or indenyl, or a ring having 10 carbon atoms ("C 10 aryl) such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms ("C 13 aryl), such as fluorenyl, or a ring having 14 carbon atoms ("C 14 aryl”), such as anthracenyl. When the C 6-20 When the aryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there is no limitation on the position of substitution, and for example, substitution may be at the ortho, para or meta position.

[0078] The term "5-14 membered heteroaryl" is 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, in particular 5 or 6 or 9 or 10 carbon atoms, and which contain 1 to 5, preferably 1 to 3, heteroatoms each independently selected from N, O and S and which, in each case, may additionally be benzofused. "Heteroaryl" also refers to a radical in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclyl rings, wherein the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 6- or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aHcarbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-carbazolylcarbazolyl, 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-, 4-, 5-, 6-, 7-, 8- or 9-piperidinyl, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthrolinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenanthrolinyl, , 3-, 4-, 5-, 6-, or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, or 10-benzoisoquinolyl, 2-, 3-, 4-, or thieno[2,3-b]furanyl, 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-oxazinyl, 1-, 3-, or 5-1H-pyrazolo[4,3-d]-oxazolyl, 2-, 4- or 5-4H-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-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzoimidazolyl, 2-, 4-, 4-, 5-, 6-, or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-4H-pyrrolo[1,2-b][2]benzazapinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. When the 5-20 membered heteroaryl group is linked to other groups to form the compounds of the present invention, the carbon atoms on the 5-20 membered heteroaryl ring may be linked to the other groups, or heteroatoms on the 5-20 membered heteroaryl ring may be linked to the other groups. When the 5-20 membered heteroaryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there is no limitation on the substitution site, for example, a hydrogen atom connected to a carbon atom on a heteroaryl ring may be substituted, or a hydrogen atom connected to a heteroatom on a heteroaryl ring may be substituted.

[0079] The term "spirocyclic" refers to a ring system in which two rings share one ring atom.

[0080] The term "fused ring" refers to a ring system in which two rings share two ring atoms.

[0081] The term "bridged ring" refers to a ring system in which two rings share three or more ring atoms.

[0082] Unless otherwise specified, a heterocyclic group, heteroaryl group, or heteroarylene group includes all possible isomeric forms thereof, such as positional isomers thereof. Thus, for some illustrative non-limiting examples, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-positions, etc. (if present) may include 1, 2, or more substituted or bonded forms thereof, including pyridin-2-yl, pyridin-2-ylene, pyridin-3-yl, pyridin-3-ylene, pyridin-4-ylene, and pyridin-4-ylene; thienyl or thienylene group includes thien-2-yl, thien-2-ylene, thien-3-ylene, and thien-3-ylene; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

[0083] The term "alkyloxy" refers to -O-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy include methoxy, ethoxy, propoxy, butoxy. Alkoxy groups may be optionally substituted or unsubstituted, and when 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, or heterocycloalkyloxy.

[0084] The term "alkylamino" refers to -NH-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkylamino include methylamino, ethylamino, propylamino, isopropylamino, butylamino, and the like.

[0085] The term "(alkyl)2amino" refers to -N-(alkyl)2, wherein alkyl is as defined above. Non-limiting examples of (alkyl)2amino include dimethylamino, methylethylamino, diethylamino, dipropylamino, methylpropylamino, diisopropylamino, dibutylamino, and the like.

[0086] "Haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

[0087] The term peptide residue refers to the oligopeptide fragment in the ADC linker, which usually serves as a substrate for enzymes in tumor cells or the microenvironment to control the release of drugs, such as GGFG, which has the structure

[0088] It will be appreciated by those skilled in the art that the compounds of formula (I) may exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they may form acid addition salts; if these compounds have an acidic center, they may form base addition salts; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they may also form internal salts.

[0089] The compounds of the present invention may exist in the form of solvates (e.g., hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0090] Depending on their molecular structure, the compounds of the present invention may be chiral and therefore may exist in various enantiomeric forms. These compounds may thus 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 separated into enantiomeric compounds by chemical or physical methods well known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from the mixture by reaction 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-phenylsulfonylproline) or various optically active camphorsulfonic acids. Chromatographic enantiomer resolution can also be advantageously performed with the aid of optically active resolving agents (e.g., dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers immobilized on silica gel). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, e.g., hexane / isopropanol / acetonitrile.

[0091] The corresponding stable isomers can be separated according to known methods, for example by extraction, filtration or column chromatography.

[0092] The term "patient" refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

[0093] The term "therapeutically effective amount" refers to that amount of an active compound or drug that will elicit the biological or medical response that a researcher, veterinarian, physician, or other clinician is seeking in a tissue, system, animal, individual, or human, and includes one or more of the following: (1) prevents disease, e.g., prevents a disease, disorder, or condition in an individual who is susceptible to the disease, disorder, or condition but who is not yet experiencing or developing the pathology or symptoms of the disease. (2) inhibits disease, e.g., inhibits the disease, disorder, or condition (i.e., prevents further development of the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition. (3) alleviates disease, e.g., alleviates the disease, disorder, or condition (i.e., reverses the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition. DETAILED DESCRIPTION

[0094] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are encompassed within the scope of protection that the present invention is intended to protect.

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

[0096] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), and tetramethylsilane (TMS) was used as the internal standard.

[0097] MS measurements were performed using an Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS liquid chromatography-mass spectrometer (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), a Waters ACQuity UPLC-QD / SQD (manufacturer: Waters, MS model: Waters ACQuity Qda Detector / Waters SQ Detector), and a THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model: THERMO Q Exactive).

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

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

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

[0101] The CombiFlash rapid preparation instrument used was Combiflash Rf200 (TELEDYNE ISCO).

[0102] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm~0.2mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm~0.5mm.

[0103] Silica gel column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0104] Average kinase inhibition rate and IC 50 The values ​​were determined using a NovoStar microplate reader (BMG, Germany).

[0105] The starting materials known in the present invention can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Darui Chemicals, and other companies.

[0106] Unless otherwise specified in the examples, all reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0107] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1 L.

[0108] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0109] The pressurized hydrogenation reaction uses a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0110] The hydrogenation reaction is usually carried out by evacuating the chamber and filling it with hydrogen, and the operation is repeated three times.

[0111] A CEM Discover-S 908860 microwave reactor was used for the microwave reaction.

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

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

[0114] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the column chromatography eluent system used for the purified compounds, and the developing solvent system for thin layer chromatography included: A: dichloromethane / methanol system, B: n-hexane / ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compounds, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid were optionally added for adjustment.

[0115] Example 1

[0116] (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]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0117] (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]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0118] Step 1: N-(3-bromo-5-fluoro-4-methoxyphenyl)acetamide 1b

[0119] Dissolve 3-bromo-5-fluoro-4-methoxyaniline 1a (20 g, 90.8 mmol) in dichloromethane (40 mL). After cooling to 0°C, slowly add acetyl chloride (14.3 g, 181.6 mmol) and triethylamine (27.6 mg, 272.4 mmol). Stir the reaction mixture at 0°C for 0.5 hours. After completion of the reaction, add water (30 mL) to the reaction mixture, extract with dichloromethane (50 mL x 3). The combined organic phases are washed with saturated brine, dried, and concentrated. The resulting residue is purified by silica gel column chromatography using eluent System B to obtain the title compound 1b (20 g, 84% yield).

[0120] MS m / z (ESI): 262.1 (M+H) + .

[0121] Step 2 (E)-4-(5-Acetylamino-3-fluoro-2-methoxyphenyl)but-3-enoic acid 1c

[0122] Compound 1b (20 g, 76.3 mmol) was dissolved in dioxane (30 mL) and water (10 mL). 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 obtain the title compound 1c (20 g, 87% yield).

[0123] MS m / z (ESI): 268.1 (M+H) + .

[0124] Step 3 4-(5-Acetylamino-3-fluoro-2-methoxyphenyl)butanoic acid 1d

[0125] 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 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, 99% yield).

[0126] MS m / z (ESI): 270.1 (M+1) + .

[0127] Step 4: N-(3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 1e

[0128] Compound 1d (20 g, 74.4 mmol) was dissolved in trifluoroacetic acid (60 mL). After cooling to 0°C, trifluoroacetic anhydride (31.24 g, 148.8 mmol) was slowly added. The reaction solution was stirred at room temperature for 7 hours. After completion of the reaction, the reaction solution was slowly poured into water (50 mL) and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated sodium bicarbonate aqueous 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 obtain the title compound 1e (9.2 g, yield: 46%).

[0129] MS m / z (ESI): 252.1 (M+1) + .

[0130] Step 5 (Z)-N-(3-fluoro-7-(hydroxyimino)-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 1f

[0131] 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 slowly added to the reaction mixture in tetrahydrofuran (10 mL). Ten minutes later, 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 by addition of saturated ammonium chloride solution and 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 obtain the title compound 1f (6 g, 51% yield).

[0132] MS m / z (ESI): 281.1 (M+1) + .

[0133] Step 6: N-(7-amino-3-fluoro-4-methoxy-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 1g

[0134] (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 2N hydrochloric acid solution (20 mL), and 10% Pd / C (1.13 g, 10.7 mmol) was added. The reaction mixture was stirred at room temperature under a hydrogen atmosphere for 5 hours. After completion of the reaction, the reaction mixture was filtered and concentrated to obtain 1 g (5 g) of crude product, which was used directly in the next step without purification.

[0135] MS m / z (ESI): 267.1 (M+1) + .

[0136] Step 7 (9H-fluoren-9-yl)methyl (8-acetylamino-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 1h

[0137] Dissolve 1g (5g, 18.8mmol) of the compound obtained in the previous step in dioxane (50mL) and adjust the pH to 7-8 with sodium bicarbonate. Then slowly add 9-fluorenylmethyl-N-succinimidyl carbonate (6.36g, 18.8mmol). Stir the reaction mixture at room temperature for 1 hour. After the reaction is complete, slowly pour the reaction mixture into water (40mL) and extract with ethyl acetate (40mL x 3). The organic phases are combined, washed with saturated brine, dried, and concentrated.

[0138] The resulting residue was purified by silica gel column chromatography with eluent System B to give the title compound 1h (5.3 g, yield: 54%).

[0139] MS m / z (ESI): 489.2 (M+1) + .

[0140] Step 8 (9H-fluoren-9-yl)methyl (8-amino-6-fluoro-5-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 1i

[0141] Compound 1h (5 g, 10.25 mmol) was dissolved in dioxane (50 mL) and 12N hydrochloric acid (10 mL). The reaction mixture was stirred at 60°C for 2 hours. After completion of the reaction, the reaction mixture was slowly poured into water (50 mL) and extracted with ethyl acetate (40 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. The resulting residue was purified by silica gel column chromatography using eluent System B to obtain the title compound 1i (3.75 g, 78% yield).

[0142] MS m / z (ESI): 447.2 (M+1) + .

[0143] 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

[0144] 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 hours. After completion of the reaction, the mixture was extracted with ethyl acetate (30 mL x 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 obtain the title compound 1k (1.8 g, 36% yield).

[0145] MS m / z (ESI): 674.2 (M+1) + .

[0146] 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 11

[0147] 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 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 obtain the title compound 1l (1.08 g, 60% yield).

[0148] MS m / z (ESI): 660.2 (M+1) + .

[0149] 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

[0150] Compound 11 (500 mg, 0.7 mmol) was dissolved in N,N-dimethylformamide (5 mL), and diethylamine (166 mg, 2.3 mmol) was added. The reaction solution was stirred at room temperature for 1 hour. After the reaction, the diethylamine in the reaction solution was removed by oil pump. The crude product 1 mL was slurried with ethyl acetate to obtain a solid that was directly used in the next reaction.

[0151] MS m / z (ESI): 438.1 (M+1) + .

[0152] 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]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-4-yl-(R)-3-hydroxybutyrate

[0153] 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 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 obtain the title compound 1n (90 mg, yield: 64%).

[0154] MS m / z (ESI): 610.2 (M+1) + .

[0155] Step 13

[0156] (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]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0157] (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]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0158] Compound 1n (90 mg, 0.15 mmol) was dissolved in methanol (5 mL), and a 1M lithium hydroxide solution (2 mL) was added at room temperature. The reaction was stirred at room temperature for 15 minutes. After the reaction, the methanol was removed by swirl, and the remaining aqueous phase was lyophilized to obtain a crude product. The crude product was then purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge C18 150*19 mm, 5 μm; mobile phase 1: water (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%).

[0159] Single configuration compound 1-1 (shorter retention time):

[0160] MS m / z (ESI): 524.2 (M+1) + .

[0161] 1 H NMR (400MHz, DMSO-d6) δ10.42 (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), 5.27-5.1 1 (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).

[0162] Single configuration compound 1-2 (longer retention time):

[0163] MS m / z (ESI): 524.2 (M+1) + .

[0164] 1H NMR (400MHz, DMSO-d6) δ10.42 (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).

[0165] Example 2

[0166] (R)-N-((1S,9S)-9-ethyl-4,9-dihydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′,6,7]indolizine[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0167] (R)-N-((1R,9S)-9-ethyl-4,9-dihydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′,6,7]indolizine[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0168] The synthetic route of Example 1 was adopted, except that the first step raw material was replaced with 3-bromo-4-methoxyaniline 2a (25 g, 0.12 mol), and purification was carried out by high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge C18 150*19 mm, 5 μm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; 15 min gradient, gradient ratio: acetonitrile phase 23%-33%, flow rate: 20 mL / min) to give the title products 2-1 (19 mg, yield: 28%) and 2-2 (30 mg, yield: 44%).

[0169] Single configuration compound 2-1 (shorter retention time):

[0170] MS m / z (ESI): 506 (M+1) + .

[0171] 1H NMR (400MHz, DMSO-d6) δ10.21 (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).

[0172] Single configuration compound 2-2 (longer retention time):

[0173] MS m / z (ESI): 506 (M+1) + .

[0174] 1 H NMR (400MHz, DMSO-d6) δ10.22 (s, 1H), 8.42-8.40 (d, 1H), 7.93-7.90 (d, 1 H), 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),2.10-1.9 7 (m, 2H), 1.86 (s, 2H), 1.23 (s, 2H), 1.08-1.06 (m, 3H), 0.87-0.85 (m, 3H).

[0175] Example 3

[0176] (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]pyrano[3′,4′,6,7]indolizine[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0177] (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]pyrano[3′,4′,6,7]indolizine[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0178] Step 1: 4-Bromo-3-fluoro-5-iodoaniline 3b

[0179] 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 completion of the reaction, water was added, the system was 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 spin-dried to give a crude product 3b (66 g).

[0180] MS m / z(ESI): 315.9, 317.9(M+1) + .

[0181] 1 H NMR (400MHz, DMSO-d6) δ7.01 (d, 1H), 6.47 (dd, 1H), 5.74 (s, 2H).

[0182] Step 2: N-(4-Bromo-3-fluoro-5-iodophenyl)acetamide 3c

[0183] Compound 3b (66 g, 208.92 mmol) was dissolved in dichloromethane (660 mL) and triethylamine (42.28 g, 417.84 mmol) was added. After the system was cooled to 0°C, acetyl chloride (19.68 g, 250.70 mmol) was slowly added dropwise. After the addition was complete, the system was stirred at room temperature for 4 hours. After the reaction was completed, the solvent was dried to obtain a crude product. The crude product was redissolved in ethyl acetate, and the pH of the system was adjusted to 2-3 using dilute hydrochloric acid. The system was then extracted with ethyl acetate, washed with saturated brine, and the organic phase was collected and dried over anhydrous sodium sulfate. The solvent was dried to obtain a crude product, which was slurried with a mixed solvent of dichloromethane / methanol (10:1) to obtain the title compound 3c (60 g, yield: 80%).

[0184] MS m / z(ESI): 357.9, 359.9(M+1) + .

[0185] 1 H NMR (400MHz, DMSO-d6) δ 10.27 (s, 1H), 7.96 (s, 1H), 7.66 (dd, 1H), 2.06 (s, 3H).

[0186] Step 3 (E)-4-(5-Acetylamino-2-bromo-3-fluorophenyl)but-3-enoic acid 3d

[0187] Compound 3c (20 g, 55.87 mmol) was dissolved in a mixed solvent of dioxane (200 mL) and water (40 mL). 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, the reaction was stirred at 100°C for 16 hours. After completion of the reaction, water and dichloromethane were added, and the system was washed 3-5 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 5-7 times with ethyl acetate. The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated to obtain the crude product, compound 3d (17 g).

[0188] MS m / z(ESI): 316.0, 317.9(M+1) + .

[0189] Step 4: 4-(5-acetylamino-2-bromo-3-fluorophenyl)butyric acid 3e

[0190] Compound 3d (8 g, 25.3 mmol) was dissolved in methanol (80 mL) and a platinum-carbon catalyst (800 mg) was added. The reaction was stirred at room temperature under a hydrogen atmosphere for two hours. After the reaction was completed, the filtrate was filtered, collected, and the solvent was dried to give a crude product 3e (8 g). The crude product was used directly in the next reaction without purification.

[0191] MS m / z(ESI): 318.0, 320.0(M+1) + .

[0192] 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).

[0193] Step 5: N-(4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 3f

[0194] 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, water was added under ice-bath conditions, and the pH of the system was adjusted to 9-10 using 15% sodium hydroxide solution. The mixture 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 evaporated to obtain compound 3f (4.8 g, yield: 53%).

[0195] MS m / z(ESI): 329.0, 331.0(M+1) + .

[0196] Step 6: (Z)-N-(4-bromo-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 3g

[0197] Potassium tert-butoxide (1.62 g, 14.4 mmol) was dissolved in a mixed solvent of tetrahydrofuran (80 mL) and tert-butanol (20 mL). A solution of compound 3f (2.15 g, 7.2 mmol) in tetrahydrofuran (20 mL) was slowly added under ice-bath. The reaction was stirred at 0°C for 10 minutes, followed by the addition of isoamyl nitrite (1.27 g, 10.8 mmol). The reaction was stirred at 0°C for 50 minutes. After completion of the reaction, dilute hydrochloric acid was added to adjust the pH of the system to 4-5. The mixture was extracted with ethyl acetate, and the organic phase was washed with saturated brine. The organic phases were collected, dried over anhydrous sodium sulfate, and the solvent was spin-dried to obtain a crude product. The crude product was slurried with methyl tert-butyl ether to obtain 3g (1.1 g, 46% yield) of the title compound.

[0198] MS m / z(ESI): 329.0, 331.0(M+1) + .

[0199] 1 H NMR (400MHz, CD3OD) δ 8.52 (d, 1H), 3.21 (dd, 2H), 3.07 (dd, 2H), 2.24 (s, 3H).

[0200] Step 7: N-(7-amino-4-bromo-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 3h

[0201] Compound 3g (500 mg, 1.52 mmol) was dissolved in dioxane (10 mL), and 1 mL of 1M hydrochloric acid and a 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 filtrate was filtered, collected, and concentrated to afford crude product 3h (500 mg), which was used directly in the next reaction.

[0202] MS m / z(ESI): 315.1, 317.1(M+1) + .

[0203] Step 8 (9H-fluoren-9-yl)methyl (8-acetylamino-5-bromo-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 3i

[0204] Compound 3h (500 mg) was dissolved in dioxane (10 mL). The pH of the filtrate from step 7 was adjusted to 8-9 using saturated sodium carbonate solution, followed by the addition of fluorenylmethyloxycarbonyl chloride (432 mg, 1.67 mmol). The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the system was 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 evaporated to obtain the crude product. The resulting residue was purified by silica gel column chromatography using eluent System B to obtain the title compound 3i (300 mg, 37% yield).

[0205] MS m / z(ESI): 537.0, 539.0(M+1) + .

[0206] Step 9 (9H-fluoren-9-yl)methyl (8-amino-5-bromo-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 3j

[0207] 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 residue was purified by silica gel column chromatography using eluent System B to afford the title compound 3j (500 mg, 77% yield).

[0208] 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

[0209] 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 was stirred at 120°C for 2 hours. After completion of the reaction, the solvent was evaporated to give the crude product. The resulting residue was purified by silica gel column chromatography using eluent System B to afford the title compound 3k (250 mg, 86% yield).

[0210] MS m / z(ESI): 722.0, 724.0(M+1) + .

[0211] 1 H NMR (400MHz, CDCl3) δ8.19-7.31 (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).

[0212] 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 31

[0213] 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 residue was purified by silica gel column chromatography using eluent System A to afford the title compound 3l (100 mg, 72% yield).

[0214] MS m / z(ESI): 500.0, 502.1(M+1) + .

[0215] Step 12

[0216] (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]pyrano[3′,4′,6,7]indolizine[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0217] (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]pyrano[3′,4′,6,7]indolizine[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0218] Compound 31 (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. After the reaction, it was purified by high performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, column: WELCH Xtimate C18 21.2*250mm, 10μm; mobile phase 1: water (0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 38%-48%, flow rate: 25mL / min) to obtain compound 3-1 (1.62mg, yield: 6%) and compound 3-2 (1.32mg, yield: 7.4%).

[0219] Single configuration compound 3-1 (shorter retention time):

[0220] MS m / z(ESI): 586.0, 588.0(M+1) + .

[0221] 1 H NMR (400MHz, CD3OD) δ8.10 (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).

[0222] Single configuration compound 3-2 (longer retention time):

[0223] MS m / z(ESI): 586.0, 588.0(M+1) + .

[0224] 1H NMR (400MHz, CD3OD) δ7.79 (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).

[0225] Example 4

[0226] (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

[0227] (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

[0228] The synthetic route of Example 3 was adopted, and 3-fluoro-5-iodoaniline (25 g, 105.5 mmol) was used as the starting material. Purification was performed by high performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, column: WELCH Xtimate C18 21.2*250 mm, 10 μm; mobile phase 1: water (0.1% FA); mobile phase 2: acetonitrile; 15 min gradient, gradient ratio: acetonitrile phase 38%-48%, flow rate: 25 mL / min) to obtain the title products 4-1 (2.5 mg, yield: 12%) and 4-2 (3.1 mg, yield: 15%).

[0229] Single configuration compound 4-1 (shorter retention time):

[0230] MS m / z (ESI): 508.1 (M+1) + .

[0231] 1H NMR (400MHz, CD3OD) δ7.65 (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).

[0232] Single configuration compound 4-2 (longer retention time):

[0233] MS m / z (ESI): 508.1 (M+1) + .

[0234] 1 H NMR (400MHz, CD3OD) δ7.57 (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).

[0235] Example 5

[0236] (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]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0237] (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]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0238] Step 1: (9H-fluoren-9-yl)methyl ((9S)-4-(difluoromethoxy)-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-1-yl)carbamate 5a

[0239] Compound 11 (200 mg, 0.30 mmol) was dissolved in DMF (5 mL), and cesium carbonate (198 mg, 0.61 mmol) and diethyl (bromodifluoromethyl)phosphonate (121 mg, 0.45 mmol) were added under ice-cooling. The reaction was stirred at 0°C for 1 hour. After completion of the reaction, water was added, and the system was extracted with ethyl acetate. The organic phase was washed with saturated brine, collected, dried over anhydrous sodium sulfate, and the solvent was spin-dried to obtain a crude product. The crude product was separated and purified by column chromatography to obtain the title compound 5a (200 mg, yield: 93%).

[0240] MS m / z (ESI): 711.2 (M+1) + .

[0241] 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

[0242] 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 solvent was evaporated under vacuum using an oil pump to obtain the crude title compound 5b (90 mg). The crude product was used directly in the next reaction without purification.

[0243] MS m / z (ESI): 488.1 (M+1) + .

[0244] Step 3

[0245] (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]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0246] (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]pyrano[3′,4′,6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxybutanamide

[0247] Crude product 5b (90 mg, 0.18 mmol) was dissolved in N,N-dimethylformamide (1 mL), and (R)-3-hydroxybutyric acid (23 mg, 0.22 mmol), 2-(7-azobenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (105 mg, 0.28 mmol) and N,N-diisopropylethylamine (48 mg, 0.37 mmol) were added. The reaction was stirred at room temperature for 1 hour. After the reaction, the solvent was dried to obtain a crude product, which was purified by high performance liquid chromatography (GILSON Prep LC with UV detector, column: Xtimate C18 250*30 mm, 10 μm; mobile phase 1: water (0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 37%-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%).

[0248] Single configuration compound 5-1 (shorter retention time):

[0249] MS m / z (ESI): 574.1 (M+1) + .

[0250] 1 H NMR (400MHz, DMSO-d6) δ8.49 (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).

[0251] Single configuration compound 5-2 (longer retention time):

[0252] MS m / z (ESI): 574.1 (M+1) + .

[0253] 1H NMR (400MHz, DMSO-d6) δ8.52 (d, 2H), 8.10-8.04 (m, 1H), 7.35 (s, 1H), 7.32 (t, 1H) 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).

[0254] Example 6

[0255] (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

[0256] (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

[0257] Step 1-Bromo-3-fluoro-2-methyl-5-nitrobenzene 6b

[0258] 2-Fluoro-1-methyl-4-nitrobenzene 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 this temperature, and the reaction was maintained at 60°C for 2 hours. The reaction solution was cooled to room temperature and added dropwise to ice water, extracted with toluene, and the organic phases were combined and washed sequentially with sodium sulfite solution, water, and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound 6b (30.0 g), which was used directly in the next reaction without purification.

[0259] MS m / z (ESI): 233.9 (M+1) + .

[0260] 1H NMR (400MHz, CDCl3) δ 8.29-8.23 (m, 1H), 7.88 (dd, 1H), 2.44 (d, 3H).

[0261] Step 2 3-Bromo-5-fluoro-4-methylaniline 6c

[0262] 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 hydrogen replacement, the mixture was reacted at room temperature under a hydrogen atmosphere for 16 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was concentrated to obtain the title compound 6c (20.0 g, yellow oil), which was used directly in the next reaction without purification.

[0263] MS m / z (ESI): 204.0 (M+1) + .

[0264] Step 3 N-(3-Bromo-5-fluoro-4-methylphenyl)acetamide 6d

[0265] 3-Bromo-5-fluoro-4-methylaniline 6c (20.0 g, 0.10 mol) was dissolved in dichloromethane (100 mL) under ice-cooling conditions. 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-cooling conditions. After completion of the reaction, water was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was then slurried in a mixed solvent of dichloromethane and petroleum ether (V / V = 10:1) to obtain the title compound 6d (10.0 g, yield: 32%).

[0266] MS m / z (ESI): 246.0 (M+1) + .

[0267] Step 4 (E)-5-(5-acetylamino-3-fluoro-2-methylphenyl)pent-4-enoic acid 6e

[0268] 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). 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 hours. After the reaction, water and dichloromethane were added, and the system 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 evaporated to obtain the title compound 6e (10 g). The product was directly used for the next reaction without purification.

[0269] MS m / z (ESI): 266.1 (M+1) + .

[0270] The subsequent synthetic route followed that of Example 3, replacing intermediate 3d with intermediate 6e (10 g, 0.04 mol). The final product was purified by high-performance liquid chromatography (HPLC) (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge C18 150*19 mm, 5 μm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 37%-47%, flow rate: 20 mL / min) to obtain the title products 6-1 (12.6 mg, yield: 14%) and 6-2 (17.2 mg, yield: 19%).

[0271] Single configuration compound 6-1 (shorter retention time):

[0272] MS m / z (ESI): 536 (M+1) + .

[0273] 1H NMR (400MHz, DMSO-d6) δ8.60-8.55(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, 3 H), 2.28-2.24(m, 4H), 2.10-1.67(m, 4H), 1.06(d, 3H), 0.85-0.82(m, 3H).

[0274] Single configuration compound 6-2 (longer retention time):

[0275] MS m / z (ESI): 536 (M+1) + .

[0276] 1 H NMR (400MHz, DMSO-d6) δ8.67-8.66(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).

[0277] Example 7

[0278] (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

[0279] (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

[0280] 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

[0281] Compound 3k (140 mg, 0.20 mmol) and potassium ethylene trifluoroborate (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. 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 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 afford the title compound 7a (80 mg, 60% yield).

[0282] MS m / z (ESI): 670.2 (M+1) + .

[0283] 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

[0284] Compound 7a (80 mg, 0.12 mmol) was dissolved in N,N-dimethylformamide (10 mL), and diethylamine (1 mL) was added. The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated in vacuo, slurried with ethyl acetate, and filtered to obtain the title compound 7b (30 mg, 51% yield).

[0285] MS m / z (ESI): 448.2 (M+1) + .

[0286] Step 3

[0287] (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

[0288] (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

[0289] Compound 7b (30 mg, 0.07 mmol) and (R)-3-hydroxybutyric acid (8 mg, 0.08 mmol) were dissolved in N,N-dimethylformamide (5 mL), and N,N-diisopropylethylamine (18.2 mg, 0.14 mmol) and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (39.9 mg, 0.11 mmol) were added. The reaction mixture was stirred at 25°C for 5 hours. After the reaction, the reaction solution was concentrated and purified by high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, chromatographic column: Xbridge C18 150*19mm, 5μm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 43%-53%, flow rate: 20mL / min) to obtain the title compound 7-1 (1.76mg, yield: 10%) and the title compound 7-2 (2.31mg, yield: 10%).

[0290] Single configuration compound 7-1 (shorter retention time):

[0291] MS m / z (ESI): 534.2 (M+1) + .

[0292] 1 H NMR (400MHz, DMSO-d6) δ8.51 (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).

[0293] Single configuration compound 7-2 (longer retention time):

[0294] MS m / z (ESI): 534.2 (M+1) + .

[0295] 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).

[0296] Example 8

[0297] (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

[0298] (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

[0299] 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

[0300] Compound 11 (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 completion of the reaction, water (10 mL) was added for dilution, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined and concentrated. The resulting residue was purified by silica gel column chromatography using eluent System B to obtain the title compound 8a (180 mg, 75% yield).

[0301] MS m / z(ESI):742.2(M+H) + .

[0302] 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

[0303] 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 obtain a crude product 8b (100 mg), which was used directly in the next reaction without purification.

[0304] MS m / z(ESI):520.1(M+H) + .

[0305] Step 3

[0306] (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

[0307] (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

[0308] Compound 8b (50 mg, 0.09 mmol) was dissolved in N,N-dimethylformamide (2 mL), and (R)-3-hydroxybutyric acid (15 mg, 0.14 mmol), 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (73 mg, 0.19 mmol), and N,N-diisopropylethylamine (37 mg, 0.28 mmol) were added. The mixture was stirred at room temperature for 1 hour. After the reaction, the product was concentrated under reduced pressure to obtain a crude product, which was prepared by high performance liquid chromatography (GILSON Prep LC with UV detector, column: Xtimate C18 150*19mm, 10μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 38%-48%, flow rate: 50mL / min) to obtain the title compound 8-1 (2.5mg, yield: 4.6%) and the title compound 8-2 (3.5mg, yield: 6.4%).

[0309] Single configuration compound 8-1 (shorter retention time):

[0310] MS m / z (ESI): 606.2 (M+1) + .

[0311] 1 H NMR (400MHz, DMSO-d6) δ8.43 (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).

[0312] Single configuration compound 8-2 (longer retention time):

[0313] MS m / z (ESI): 606.2 (M+1) + .

[0314] 1H NMR (400MHz, DMSO-d6) δ8.45 (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(d d, 1H), 2.09 (dd, 2H), 1.93-1.78 (m, 2H), 1.09 (d, 3H), 0.94-0.78 (m, 3H).

[0315] Example 9

[0316] (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

[0317] Step 1 8-amino-5-methoxy-3,4-dihydronaphthalen-1(2H)-one 9a

[0318] 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 obtain the title compound 9a (400 mg, yield: 88%).

[0319] MS m / z (ESI): 192.1 (M+1) + .

[0320] 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

[0321] 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, and the mixture was extracted with dichloromethane (15 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. Purification by silica gel column chromatography using eluent System B afforded the title compound 9b (698 mg, 80% yield).

[0322] MS m / z (ESI): 419.1 (M+1)+ .

[0323] Step 3 (S)-9-ethyl-4,9-dihydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzopyrano[3′,4′,6,7]indolizine[1,2-b]quinoline-10,13-dione 9

[0324] Compound 9b (100 mg, 0.24 mmol) was dissolved in hydrobromic acid (30 mL). The reaction mixture was reacted at 100°C for 2 hours. After the reaction, the reaction mixture was directly concentrated and purified by high-performance liquid chromatography (Waters MS triggered Prep-LC with Acquity QDA detector, column: Welch C18 250×21.2 mm, 10 μm; mobile phase 1: water (containing 0.1% ammonia water); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 10%-30%, flow rate: 25 mL / min) to obtain the title compound 9 (19.4 mg, 20%).

[0325] MS m / z (ESI): 405.1 (M+1) + .

[0326] 1 H NMR (400MHz, CD3OD) δ7.79-7.71 (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).

[0327] Example 10

[0328] (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

[0329] first step

[0330] (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

[0331] (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

[0332] Compound 6m (700 mg) was prepared by high performance liquid chromatography (GILSON Prep LC with UV detector, column: Xbridge 5μm C18 150x30mm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 22%-32%, flow rate: 50 mL / min) to obtain compounds 6m-1 (290 mg, shorter retention time) and 6m-2 (300 mg, longer retention time).

[0333] MS m / z (ESI): 450.2 (M+H) +

[0334] Step 2

[0335] (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

[0336] Compound 6m-2 (30 mg, 0.06 mmol) was dissolved in N,N-dimethylformamide (3 mL), and (1S,3S)-3-hydroxycyclobutane-1-carboxylic 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 solution was stirred at room temperature for 1 hour. After completion of the reaction, water (10 mL) was added to dilute the reaction solution, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated to obtain a crude product. The crude product was purified by HPLC (GILSON Prep LC with UV detector, column: Xbridge 5 μm C18 150x30 mm; mobile phase 1: water (containing 0.1% TFA); mobile phase 2: acetonitrile; 15-min gradient, gradient ratio: acetonitrile phase 30%-95%, flow rate: 50 mL / min) to give compound 10 (2 mg, yield: 5%).

[0337] MS m / z(ESI):548.2(M+H) + .

[0338] 1 H NMR (400MHz, CD3OD) δ8.71 (d, 1H), 7.57-7.48 (m, 2H), 5.65 (s, 1H), 5.55 (d, 1H), 5.35 (dd, 2H), 5.10 (d, 1 H), 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).

[0339] Example 11

[0340] (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

[0341] (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

[0342] Step 1: 2-cyclopropyl-2-hydroxyacetic acid 11b

[0343] 2-cyclopropyl-2-hydroxyacetic acid methyl ester 11a (50 mg, 0.38 mmol) is dissolved in tetrahydrofuran (6 mL), and water (0.5 mL) and lithium hydroxide (27 mg, 1.15 mmol) are added. The reaction is stirred at room temperature for 1 hour. After the reaction is completed, the solvent is spin-dried to obtain a crude product 2-cyclopropyl-2-hydroxyacetic acid 11b (50 mg, white solid), which is directly used for the next step reaction without purification.

[0344] MS m / z (ESI): 117.1 (M+H) + .

[0345] Step 2

[0346] (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

[0347] (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

[0348] Compound 6m-2 (50 mg, 0.11 mmol) was dissolved in N,N-dimethylformamide (2 mL), and 2-cyclopropyl-2-hydroxyacetic acid 11b (44 mg, 0.38 mmol), N,N-diisopropylethylamine (29 mg, 0.22 mmol) and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (63 mg, 0.17 mmol) were added. The reaction was stirred at room temperature for 15 minutes. After the reaction, the product was purified by high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5μm C18 150x19mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 10 min gradient, gradient ratio: acetonitrile phase 40%-50%, flow rate: 20 mL / min) was purified to obtain compound 11-1 (2.3 mg, yield: 4%) and compound 11-2 (2.5 mg, yield: 4%).

[0349] Single configuration compound 11-1 (compound with shorter retention time)

[0350] MS m / z(ESI):548.2(M+H) + .

[0351] 1 H NMR (400MHz, DMSO-d6) δ8.49 (d, 1H), 7.74 (d, 1H), 7.28 (s, 1H), 6.50 (s, 1H), 5.56 (s, 1H), 5.49-5.40 (m, 4H), 5.2 9 (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).

[0352] Single configuration compound 11-2 (compound with longer retention time)

[0353] MS m / z(ESI):548.2(M+H) + .

[0354] 1H NMR (400MHz, DMSO-d6) δ8.43 (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, 1 H), 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).

[0355] Example 12

[0356] (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

[0357] 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

[0358] (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

[0359] Compound 7b (500 mg) was prepared by high performance liquid chromatography (GILSON Prep LC with UV detector, column: Xbridge 5μm C18 150x30mm; mobile phase 1: water (0.1% TFA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 22%-32%, flow rate: 50 mL / min) to give compounds 7b-1 (180 mg, shorter retention time) and 7b-2 (195 mg, longer retention time).

[0360] MS m / z(ESI):448.2(M+H) + .

[0361] 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

[0362] Compound 7b-2 (22 mg, 0.05 mmol) was dissolved in N,N-dimethylformamide (0.5 mL), and (1S,3S)-3-hydroxycyclobutane-1-carboxylic acid (7 mg, 0.06 mmol), 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (30 mg, 0.08 mmol) and N,N-diisopropylethylamine (13 mg, 0.10 mmol) were added. The reaction was stirred at room temperature for 15 minutes. After the reaction, the reaction solution was purified by high-performance liquid chromatography (HPLC) (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5 μm C18 150x19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 10-min gradient, gradient ratio: acetonitrile phase 45%-100%, flow rate: 20 mL / min) to obtain compound 12 (3 mg, yield: 11%).

[0363] MS m / z (ESI): 546.2 (M+H) + .

[0364] 1 H NMR (400MHz, CD3OD) δ8.50 (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).

[0365] Example 13

[0366] (R)-N-((1R,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-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

[0367] (R)-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-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

[0368] The first step is N-(4-cyclopropyl-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 17a

[0369] Compound 3f (1.5 g, 5.02 mmol), cyclopropylboronic acid (1.3 g, 15.06 mmol), 1,1-bis(diphenylphosphino)diboronium palladium chloride (2.2 g, 3.01 mmol), and cesium carbonate (1.6 g, 15.06 mmol) were dissolved in dioxane (15 mL). The reaction was stirred in a microwave reactor at 110°C for 2 hours. After completion of the reaction, the reaction solution 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 afforded the title compound 17a (1.1 g, 84% yield).

[0370] MS m / z (ESI): 262.1 (M+1) + .

[0371] Step 2 (Z)-N-(4-cyclopropyl-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 17b

[0372] Potassium tert-butoxide (0.94 g, 8.4 mmol) was dissolved in a mixed solvent of tetrahydrofuran and tert-butanol (50 mL, V / V = 4:1). Compound 17a (1.1 g, 4.2 mmol) was dissolved in 10 mL of tetrahydrofuran solution and slowly added to the reaction system at 0°C. The reaction was stirred at 0°C for 10 minutes, and then isoamyl nitrite (0.74 g, 6.3 mmol) was added. The reaction was stirred at 0°C for 50 minutes. After completion of the reaction, dilute hydrochloric acid was added to adjust the pH of the system to 4-5, and the mixture was extracted three times with ethyl acetate. The organic phase was washed with saturated brine, collected, dried over anhydrous sodium sulfate, and the solvent was spin-dried to obtain the crude product 17b, which was used directly in the next reaction without purification.

[0373] MS m / z (ESI): 291.0 (M+1) + .

[0374] Step 3: N-(7-amino-4-cyclopropyl-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide 17c

[0375] Compound 17b was dissolved in methanol (50 mL) and palladium-carbon catalyst (500 mg) was added. The reaction was stirred at room temperature under a hydrogen atmosphere for two hours. After completion of the reaction, the filtrate was filtered and collected to obtain crude product 17c, which was used directly in the next reaction without further purification.

[0376] MS m / z (ESI): 277.1 (M+1) + .

[0377] Step 4 (9H-fluoren-9-yl)methyl (8-acetylamino-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 17d

[0378] The filtrate obtained in the previous step was adjusted to a pH of 8-9 using saturated sodium carbonate solution, followed by the addition of Fmoc-Cl (1.19 g, 4.6 mmol). The reaction was stirred at room temperature for 1 hour. After completion of the reaction, the system was 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 evaporated to obtain a crude product. The crude product was purified by silica gel column chromatography System A to obtain compound 17d (1 g, yield: 48%).

[0379] MS m / z (ESI): 499.1 (M+1) + .

[0380] Step 5 (9H-fluoren-9-yl)methyl (8-amino-5-cyclopropyl-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate 17e

[0381] 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 completion of the reaction, the solvent was evaporated to obtain a crude product, which was purified using silica gel column chromatography System A to obtain compound 17f (700 mg, 77% yield).

[0382] MS m / z (ESI): 457.1 (M+1) + .

[0383] Step 6: (9H-fluoren-9-yl)methyl((9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)carbamate 17g

[0384] 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 system was stirred at 140°C for 4 hours. After completion of the reaction, the solvent was evaporated to obtain a crude product, which was purified by silica gel column chromatography (Achu) to obtain the title compound 17g (700 mg, 66% yield).

[0385] MS m / z (ESI): 684.2 (M+1) + .

[0386] 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

[0387] 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 afford the crude product, Compound 17h (60 mg). The crude product was carried on to the next step without further purification.

[0388] MS m / z (ESI): 462.1 (M+1) + .

[0389] Step 8 (R)-N-((1R,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-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

[0390] (R)-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-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

[0391] 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 the reaction, the reaction solution was purified by high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5μm C18 150x19mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 38%-48%, flow rate: 20mL / min) to obtain compound 17-1 (6 mg, yield: 11%) and 17-2 (4 mg, yield: 8%). Single configuration compound 17-1 (shorter retention time):

[0392] MS m / z (ESI): 548.2 (M+1) + .

[0393] 1 H NMR (400MHz, DMSO-d6) δ8.44(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).

[0394] Single configuration compound 17-2 (longer retention time):

[0395] MS m / z (ESI): 548.2 (M+1) + .

[0396] 1H 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, 1 H), 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).

[0397] Example 14

[0398] (1S,3S)-N-((1R,9S)-4-Cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-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

[0399] (1S,3R)-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-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

[0400] first step

[0401] (1S,3S)-N-((1R,9S)-4-Cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-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

[0402] (1S,3R)-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-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

[0403] Compound 17g (69 mg, 0.15 mmol) was dissolved in N,N-dimethylformamide (5 mL) and (1S,3S)-3-hydroxycyclobutane-1-carboxylic acid (21 mg, 0.18 mmol), 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (87 mg, 0.23 mmol) and N,N-dimethylacetamide (59 mg, 0.45 mmol) were added. The reaction was stirred at room temperature for 30 minutes. After the reaction, the reaction solution was purified by high performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, column: WELCH Xtimate C18 21.2x250mm 10μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 18-minute gradient, gradient ratio: acetonitrile phase 35%-65%, flow rate: 20mL / min) to obtain 18-1 (4 mg, yield: 6%) and 18-2 (2.4 mg, yield: 4%).

[0404] Single configuration compound 18-1 (shorter retention time):

[0405] MS m / z (ESI): 560.2 (M+1) + .

[0406] 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).

[0407] Single configuration compound 18-2 (longer retention time):

[0408] MS m / z (ESI): 560.2 (M+1) + .

[0409] 1H NMR (400MHz, DMSO-d6) δ8.45 (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).

[0410] Example 15

[0411] (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

[0412] (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

[0413] Step 1: 5-(5-acetylamino-2-bromo-3-fluorophenyl)pent-4-enoic acid 19b

[0414] Compound 19a (7.0 g, 19.5 mmol) was dissolved in dioxane (60 mL) and water (15 mL), and pent-4-enoic acid (2.2 g, 22.0 mol), bistriphenylphosphine palladium dichloride (700 mg, 1.0 mmol), and sodium carbonate (6.2 g, 58.4 mmol) were added to the solution in sequence. The reaction solution was stirred at 90 ° C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was filtered through celite and washed with ethyl acetate, and the filtrate was directly concentrated. The crude product was diluted with water (200 mL) and extracted with ether three times (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). MS m / z (ESI): 330.1 (M+1) + .

[0415] Step 2 5-(5-Acetylamino-2-bromo-3-fluorophenyl)pentanoic acid 19c

[0416] Compound 19b (6.0 g, 18.2 mmol) was dissolved in anhydrous methanol (100 mL), and platinum / carbon (5%, 600 mg) was added. The reaction solution was replaced with hydrogen three times and stirred at room temperature for 16 hours. After completion of the reaction, the reaction solution was filtered through celite and washed with ethyl acetate. The resulting filtrate was directly concentrated to obtain the crude product, compound 19c (5.0 g).

[0417] MS m / z (ESI): 332.0 (M+1) + .

[0418] Step 3: N-(4-bromo-3-fluoro-9-oxo-6,7,8,9-tetrahydro-5H-benzo[7]cycloalken-1-yl)acetamide 19d

[0419] 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 completion of the reaction, 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 to obtain a crude product which was purified by silica gel column chromatography system B to afford compound 19d (920 mg, 39% yield).

[0420] MS m / z (ESI): 314.0 (M+1) + .

[0421] The subsequent synthetic route was the same as that of Example 17, except that compound 3f was replaced with 19d (720 mg, 2.3 mmol). The final product was purified by high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, column: WELCH Xtimate C18 21.2x250 mm 10 μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-min gradient, gradient ratio: acetonitrile phase 40%-95%, flow rate: 30 mL / min) to obtain products 19-1 (6.5 mg, yield: 9%) and 19-2 (11 mg, yield: 16%).

[0422] Single configuration compound 19-1 (shorter retention time):

[0423] MS m / z (ESI): 562.2 (M+1) + .

[0424] 1H NMR (400MHz, CDCl3) δ7.63-7.50 (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).

[0425] Single configuration compound 19-2 (longer retention time):

[0426] MS m / z (ESI): 562.2 (M+1) + .

[0427] 1 H NMR (400MHz, CDCl3) δ7.50-7.35(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).

[0428] Example 16

[0429] (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

[0430] (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

[0431] first step

[0432] (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

[0433] (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

[0434] Compound 19k (75 mg, 0.16 mmol) was dissolved in N,N-dimethylformamide (3 mL), and (1S,3S)-3-hydroxycyclobutane-1-carboxylic 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, the reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). The organic phases were combined and washed with saturated brine, dried, and concentrated. The crude product was purified by high performance liquid chromatography (GILSON Prep LC with UV detector, column: WELCH Xtimate C18 21.2x250mm 10μm; mobile phase 1: water (containing 0.1% NH3.H20); mobile phase 2: acetonitrile; 15 min gradient, gradient ratio: acetonitrile phase 40%-100%, flow rate: 30 mL / min) to give compound 20-1 (2.9 mg, 7%) and 20-2 (1.5 mg, 2%).

[0435] Single configuration compound 20-1 (shorter retention time):

[0436] MS m / z (ESI): 574.7 (M+1) + .

[0437] 1H NMR (400MHz, CDCl3) δ7.59 (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).

[0438] Single configuration compound 20-2 (longer retention time):

[0439] MS m / z (ESI): 574.7 (M+1) + .

[0440] 1 H NMR (400MHz, CDCl3) δ7.53 (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),3.6 8(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).

[0441] Example 17

[0442] (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

[0443] (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

[0444] First step (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

[0445] Compound 19j (500 mg, 0.72 mmol) was purified using a Gilson instrument and a Daicel chiral column to separate the chiral isomers (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 obtain compound 23a (220 mg, yield: 44%). 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

[0446] 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 stirred at room temperature for 2 hours. After the reaction, the solution was concentrated to afford crude compound 23b (30 mg), which was used directly in the next reaction without purification.

[0447] MS m / z (ESI): 476.2 (M+1) + .

[0448] Step 3

[0449] (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

[0450] (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

[0451] 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 mixture was stirred at room temperature for 3 hours. After the reaction, the reaction solution was purified by high performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, column: WELCH Xtimate C18 21.2 x 250mm 10μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-min gradient, gradient ratio: acetonitrile phase 46%-100%, flow rate: 30mL / min) to obtain products 33-1 (4.1 mg, yield: 17.2%) and 33-2 (1.7 mg, yield: 6.9%).

[0452] 33-1 (single configuration compound, short retention time)

[0453] MS m / z (ESI): 574.2 (M+1) + .

[0454] 1 H NMR (400MHz, DMSO-d6) δ8.62-8.37(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), 3.49-3.4 6(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).

[0455] 33-2 (single configuration compound, longer retention time)

[0456] MS m / z (ESI): 574.2 (M+1) + .

[0457] 1 H NMR (400MHz, DMSO-d6) δ8.45 (d, 2H), 7.69 (d, 1H), 7.28 (s, 1H), 6.53 (s, 1H), 5. 60-5.52(m,1H),5.48(s,1H),5.43(d,2H),5.30(d,1H),3.59(d,1H),3.49-3.4 6(m, 1H), 2.14-1.79(m, 6H), 1.75-1.61(m, 1H), 1.24(s, 1H), 1.17-1.02(m, 3H) , 0.86 (t, J=8Hz, 3H), 0.83-0.77 (m, 1H), 0.74-0.64 (m, 1H), 0.48-0.29 (m, 4H).

[0458] Example 18

[0459] (S)-3-Cyclopropyl-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypropionamide

[0460] (R)-3-Cyclopropyl-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypropionamide

[0461] Step 1: (9H-fluoren-9-yl)methyl((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-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

[0462] Compound 17f (600 mg, 1.02 mmol) was purified using a Gilson instrument and a Daicel chiral column to separate the chiral isomers (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 obtain compound 21a (250 mg, yield: 42%). 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

[0463] 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 afford the title compound 21b (34 mg), which was used directly in the next reaction without further purification.

[0464] MS m / z (ESI): 462.1 (M+1) + .

[0465] Step 3

[0466] (S)-3-Cyclopropyl-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypropionamide

[0467] (R)-3-Cyclopropyl-N-((1S,9S)-4-cyclopropyl-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypropionamide

[0468] Compound 21b (30 mg, 0.065 mmol) was dissolved in N,N-dimethylformamide (3 mL), and 3-cyclopropyl-3-hydroxypropionic acid (15 mg, 0.098 mmol), 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (37 mg, 0.096 mmol), and N,N-diisopropylethylamine (171 mg, 1.32 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction, the product 34-1 (4.2 mg, yield: 11%) and 34-2 (3.9 mg, yield: 10%) were purified by high performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector, 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%-100%, flow rate: 30 mL / min).

[0469] 34-1 (single configuration compound, compound with short retention time)

[0470] MS m / z (ESI): 574.2 (M+1) + .

[0471] 1 H NMR (400MHz, DMSO-d6) δ8.53-8.39 (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, 2 H), 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).

[0472] 34-2 (single configuration compound, compound with longer retention time)

[0473] MS m / z (ESI): 574.2 (M+1) + .

[0474] 1H NMR (400MHz, DMSO-d6) δ8.45 (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).

[0475] Example 19

[0476] (R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypentanamide

[0477] (S)-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-hydroxypentanamide

[0478] first step

[0479] (R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxypentanamide

[0480] (S)-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-hydroxypentanamide

[0481] Compound 6m-2 (30 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (3 mL), and 3-hydroxyvaleric acid (9 mg, 0.07 mmol), N,N-diisopropylethylamine (17 mg, 0.13 mmol), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (38 mg, 0.10 mmol) were added. The reaction system was reacted at room temperature for 1 hour. After the reaction, high performance liquid chromatography (HPLC) was used (GILSON Prep LC with UN detector, 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 phase 5%-57%, flow rate: 50 mL / min) to purify compounds 36-1 (6.98 mg, yield: 18%) and 36-2 (6.70 mg, yield: 18%).

[0482] 36-1 (single configuration compound, compound with short retention time)

[0483] MS m / z (ESI): 550.2 (M+1) + .

[0484] 1 H NMR (400MHz, DMSO-d6) δ8.61 (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, 1 H), 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).

[0485] 36-2 (single configuration compound, compound with longer retention time)

[0486] MS m / z (ESI): 550.2 (M+1) + .

[0487] 1H 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).

[0488] Example 20

[0489] (S)-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-4-methylpentanamide

[0490] (R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxy-4-methylpentanamide

[0491] first step

[0492] (S)-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-4-methylpentanamide

[0493] (R)-N-((1S,10S)-10-ethyl-6-fluoro-10-hydroxy-5-methyl-11,14-dioxo-1,2,3,4,10,11,14,16-octahydro-13H-cyclohepta[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-3-hydroxy-4-methylpentanamide

[0494] Compound 6m-2 (30 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (3 mL), and 3-hydroxy-4-methylpentanoic acid (9 mg, 0.07 mmol), N,N-diisopropylethylamine (17 mg, 0.13 mmol), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (38 mg, 0.10 mmol) were added. The reaction system was reacted at room temperature for 1 hour. After the reaction, high performance liquid chromatography (HPLC) was used (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5μm C18 150 x 19mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 42%-100%, flow rate: 20 mL / min) to purify compounds 37-1 (6.6 mg, yield: 18%) and 37-2 (6.9 mg, yield: 18%).

[0495] 37-1 (single configuration compound, compound with short retention time)

[0496] MS m / z (ESI): 564.2 (M+1) + .

[0497] 1 H NMR (400MHz, DMSO-d6) δ8.68 (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).

[0498] 37-2 (single configuration compound, compound with longer retention time)

[0499] MS m / z (ESI): 564.2 (M+1) + .

[0500] 1H NMR (400MHz, DMSO-d6) δ8.78 (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).

[0501] Example 21

[0502] (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

[0503] (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

[0504] (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

[0505] (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

[0506] Compound 6m-2 (30 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (3 mL), and 3-cyclopropyl-3-hydroxypropionic acid (18 mg, 0.14 mmol), N,N-diisopropylethylamine (17 mg, 0.13 mmol), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (38 mg, 0.10 mmol) were added. The reaction system was reacted at room temperature for 1 hour. After the reaction, high performance liquid chromatography (HPLC) was used (Waters MS-triggered Prep-LC with QDA detector, column: Xbridge 5μm C18 150 x 19mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-minute gradient, gradient ratio: acetonitrile phase 45%-100%, flow rate: 25 mL / min) to purify and separate the title compound 38-1 (4 mg, yield: 11%) and the title compound 38-2 (4 mg, yield: 11%).

[0507] 38-1 (single configuration compound, compound with short retention time)

[0508] MS m / z (ESI): 562.3 (M+1) + .

[0509] 1 H NMR (400MHz, DMSO-d6) δ8.63 (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, 2 H), 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).

[0510] 38-2 (single configuration compound, compound with longer retention time)

[0511] MS m / z (ESI): 562.3 (M+1) + .

[0512] 1H NMR (400MHz, DMSO-d6) δ8.71 (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), 3.2 2(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).

[0513] Example 22

[0514] (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

[0515] first step

[0516] (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

[0517] Compound 6m-2 (30 mg, 0.07 mmol) was dissolved in N,N-dimethylformamide (2 mL), and (2S,3R)-3-hydroxy-2-methylbutanoic 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 product was purified by preparative HPLC (Waters MS-triggered Prep-LC with QDA detector, column: Xbridge 5μm C18 150 x 19 mm, 5μm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 10-minute gradient, gradient ratio: acetonitrile 42% to 52%, flow rate: 25 mL / min) to obtain compound 39 (2 mg, yield: 5%).

[0518] MS m / z (ESI): 550.0 (M+1) + .

[0519] 1 H NMR (400MHz, CDCl3) δ7.73 (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).

[0520] Biological evaluation

[0521] The present invention is further described and explained below in conjunction with the test examples, but these test examples are not intended to limit the scope of the present invention. Test Example 1 Inhibition experiment of compound on SK-BR-3 cell proliferation

[0522] 1.1 Experimental Materials

[0523] 1.2 Experimental instruments

[0524] 1.3 Test Method

[0525] (1) Cell plating: First, culture the tumor cells SK-BR-3 in the corresponding culture medium, digest the cells with trypsin, resuspend the cells after centrifugation, count the cells, adjust the cells to the appropriate concentration, and plate them on a 384-well plate.

[0526] (2) Co-incubation of compounds and tumor cells: After cells adhered to the wall, 100 nL of diluted bioactive substance (test compound) was added to the cell culture plate using ECHO. The final DMSO concentration in the cell plate wells was 0.33%. The cells were incubated in a 37°C, 5% CO2 incubator for 72 h.

[0527] (3) After the incubation, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the wells were shaken on a rapid shaker for 2 minutes, centrifuged at 1000 rpm for 1 minute, and placed in the dark at room temperature for 30 minutes. The chemiluminescence signal value was read using an Envision instrument.

[0528] (4) Cell viability assay: IC was calculated using GraphPad Prism 8 software 50 , the IC of the compound is obtained using the following nonlinear fitting formula 50 (See Table 1):

[0529] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0530] Y: inhibition rate; X: log value of compound concentration;

[0531] Inhibition rate (%) = 100 - (reading value of compound well - reading value of low reading control well) / (reading value of high reading control well - reading value of low reading control well) * 100;

[0532] High-reading control wells: cells plus 100 nL DMSO;

[0533] Low reading control wells: wells without cells.

[0534] Table 1: Inhibitory activity of test compounds on SK-BR-3 proliferation

[0535] The results showed that the compound of the present invention has a strong inhibitory effect on the proliferation of SK-BR-3 cells.

[0536] Test Example 2 Inhibitory experiment of test compound on MDA-MB-468 cell proliferation

[0537] 2.1 Experimental Materials

[0538] 2.2 Experimental instruments

[0539] 2.3 Test Method

[0540] (1) Cell plating: First, culture the tumor cells MDA-MB-468 in the corresponding culture medium, digest the cells with trypsin, resuspend the cells after centrifugation, count the cells, adjust the cells to the appropriate concentration, and plate them on a 384-well plate.

[0541] (2) Co-incubation of compounds and tumor cells: After cells adhered to the wall, 100 nL of diluted bioactive substance (test compound) was added to the cell culture plate using ECHO. The final DMSO concentration in the cell plate wells was 0.33%. The cells were incubated in a 37°C, 0% CO2 incubator for 72 h.

[0542] (3) After the incubation, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the wells were shaken on a rapid shaker for 2 minutes, centrifuged at 1000 rpm for 1 minute, and placed in the dark at room temperature for 30 minutes. The chemiluminescence signal value was read using an Envision instrument.

[0543] (4) Cell viability assay: IC was calculated using GraphPad Prism 8 software 50 , the IC of the compound is obtained using the following nonlinear fitting formula 50 (See Table 2):

[0544] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0545] Y: inhibition rate; X: log value of compound concentration;

[0546] Inhibition rate (%) = 100 - (reading value of compound well - reading value of low reading control well) / (reading value of high reading control well - reading value of low reading control well) * 100;

[0547] High read value control well: cells plus 100nL DMSO

[0548] Low reading control wells: wells without cells.

[0549] Table 2: Antiproliferation activity of test compounds on MDA-MB-468

[0550] The results showed that the test compound had a strong inhibitory effect on the proliferation of MDA-MB-468 cells.

[0551] Test Example 3 Inhibitory Effect of Test Compounds on NCI-N87 Cell Proliferation

[0552] 3.1 Experimental Materials

[0553] 3.2 Experimental instruments

[0554] 3.3 Test Method

[0555] (1) Cell plating: First, culture the tumor cells NCI-N87 in the corresponding culture medium, digest the cells with trypsin, resuspend the cells after centrifugation, count the cells, adjust the cells to an appropriate concentration, and plate them on a 384-well plate.

[0556] (2) Co-incubation of compounds and tumor cells: After cells adhered to the wall, 100 nL of diluted bioactive substance (test compound) was added to the cell culture plate using ECHO. The final DMSO concentration in the cell plate wells was 0.33%. The cells were incubated in a 37°C, 0% CO2 incubator for 72 h.

[0557] (3) After the incubation, 30 μL of CTG reagent (CelltiterGlo kit) was added to each well, and the wells were shaken on a rapid shaker for 2 minutes, centrifuged at 1000 rpm for 1 minute, and placed in the dark at room temperature for 30 minutes. The chemiluminescence signal value was read using an Envision instrument.

[0558] (4) Cell viability assay: IC was calculated using GraphPad Prism 8 software 50 , the IC of the compound is obtained using the following nonlinear fitting formula 50 (See Table 3): Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope))

[0559] Y: inhibition rate; X: log value of compound concentration;

[0560] Inhibition rate (%) = 100 - (reading value of compound well - reading value of low reading control well) / (reading value of high reading control well - reading value of low reading control well) * 100;

[0561] High-reading control wells: cells plus 100 nL DMSO;

[0562] Low reading control wells: wells without cells.

[0563] Table 3 Inhibitory activity of test compounds on NCI-N87 cell proliferation

[0564] The results showed that the test compound had a strong inhibitory effect on the proliferation of NCI-N87 cells.

[0565] Test Example 4 Kinetic Solubility Test in PBS 7.4 Buffer

[0566] 4.1 Experimental steps

[0567] 1) Preparation of stock solution

[0568] 10 mM stock solutions of the test substances 6-2, 11-2, 17-2, Dxd, and the control drugs progesterone and diclofenac were prepared in DMSO.

[0569] 2) Kinetic solubility determination steps

[0570] 15 μL of 10 mM stock solution of the analyte to be tested was added to the corresponding positions of the 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 added to each vial and the vials were capped. The sample plate was then placed in a thermomixer and shaken at 1100 rpm at 25°C for 2 hours. After 2 hours, the vials were removed from the vials, the stir bars were removed using a large magnet, and the samples were transferred from the sample plate to the filter plate. A vacuum pump was used to generate negative pressure to filter the samples. 5 μL of the filtrate and 5 μL of blank DMSO were transferred to a new sample plate, and then 490 μL of internal standard water (acetonitrile:water = 1:1) containing the internal standard was added. Depending on the peak shape, the sample diluent may be diluted with a certain proportion of internal standard water to achieve better peak shape.

[0571] 3) Preparation of 3 μM standard solution

[0572] Transfer 6 μL from the 10 mM DMSO stock solution plate to an empty plate and add 194 μL of DMSO to prepare a 300 μM standard solution. Transfer 5 μL from the 300 μM standard solution plate to another empty plate and add 5 μL of blank buffer and 490 μL of internal standard water (acetonitrile:water = 1:1) to a final concentration of 3 μM standard solution.

[0573] 4) Sample analysis steps

[0574] The sample plate was placed into the sample tray of the autosampler and the samples were evaluated by liquid chromatography-mass spectrometry (LC-MS).

[0575] 4.0 Data Analysis

[0576] All calculations were performed using Microsoft Excel.

[0577] The analysis and quantification of the sample filtrate is completed by using a liquid chromatography mass spectrometry to determine the qualitative and quantitative values ​​of the standard peaks of known concentrations. The solubility values ​​of the control drug and the test substance are calculated as follows:

[0578] [Sample] is the concentration of the compound, DF Sample Refers to the sample dilution multiple, [STD] is the concentration of the compound standard, AREA Std AREA is the peak area of ​​the standard compound. Sample is the peak area of ​​the compound sample.

[0579] 4.2 Experimental Results

[0580] The experimental results are shown in Table 4.

[0581] Table 4 Solubility of representative compounds in PBS 7.4 buffer

[0582] The results showed that the solubility of the representative compounds 6-2 and 11-2 in PBS 7.4 buffer was significantly better than that of Dxd.

[0583] Test Example 5 SD rat pharmacokinetics test

[0584] After a single intravenous injection (dose of 2 mpk) of compounds 6-2, 11-2, 17-2, and Dxd into male SD rats (blood collection time points: 8 blood collection time points, 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, and 24 h after IV administration), the average pharmacokinetic parameters in plasma are shown in Table 5:

[0585] Table 5 Pharmacokinetic test results of representative compounds

[0586] The results showed that the representative compounds of the present application were rapidly cleared from rats, had good safety, and had good pharmacokinetic properties.

[0587] Test Example 6 Inhibition experiment of compounds on human liver microsome CYP1A2, CYP2D6 and CYP3A4

[0588] 6.1 Positive control inhibitors for each subtype

[0589] 6.2 Preparation of substrate stock solution

[0590] 6.3 Experimental Procedure

[0591] Incubations were performed in 96-deep-well plates. The following volumes were added to each well of the plate: 169 μL of the "master solution" and 1 μL of various concentrations of the test compound or positive control compound (DMSO). The plate was preincubated in a waterbath at 37°C for 5 minutes. Then, 10 μL of the diluted substrate solution was added to the plate, mixed on a vortex mixer for 15 seconds, and the reaction was initiated by adding 20 μL of a 10 mM NADPH solution to a final concentration of 1 mM. At predetermined time points, the reaction was quenched by adding 300 μL of a quenching solution (cold acetonitrile with 3% formic acid, 200 nM alprazolam, 200 nM labetalol hydrochloride, and 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 was diluted with 150 μL of purified water. The mixture was mixed thoroughly, and the substrate metabolite content was determined by LC / MS / MS.

[0592] 6.4 Data Analysis

[0593] An automated peak integration area check was performed on all samples. Analytical peak areas and internal standard peak areas were exported to an Excel spreadsheet. Inhibition of each P450 enzyme in human liver microsomes was measured as the percentage decrease in marker metabolite formation activity compared to a non-inhibited control (=100% activity). IC values ​​were calculated as the residual activity (%) and the logarithm of the inhibitor concentration. 50 value.

[0594] The percentage of residual activity was calculated as follows: Area ratio = analyte peak area / internal standard peak area Residual activity (%) = Area ratio 待测药 / Area ratio 空白对照 *100%

[0595] IC calculation using Excel XLfit 5.5.1.3 50 value.

[0596] The experimental results are shown in Table 6

[0597] Table 6 Inhibitory test results of representative compounds on human liver microsomes

[0598] The results showed that the representative compounds of this application had weak inhibitory effects on CYP enzymes and had good safety.

[0599] Test Example 7 Stability Test of Compounds in Human Plasma

[0600] 7.1 Experimental Procedure

[0601] (1) Add 199 μl of human plasma to each cell culture plate, preheat the culture plate to 37°C, and keep warm for 15 minutes.

[0602] (2) After preincubation, 1 μL of 1 mmol test compound and 1 μL of 1 mmol control compound were added to 199 μL of plasma to achieve a final concentration of 5 μM test compound and 5 μM control compound. The final concentration of the organic solvent was 0.5%. This experiment was repeated twice.

[0603] (3) The reaction samples were incubated at 37°C.

[0604] (4) The reaction was stopped at 0, 1, 2, 6, and 24 hours by adding 600 μL of cold methanol containing an internal standard. All samples were vortexed for 10 minutes and then centrifuged at 3220 g for 30 minutes to precipitate the protein. 100 μL of the supernatant was transferred to a new plate. Based on the signal response and peak shape of the liquid chromatography-mass spectrometry (LC-MS), the supernatant was diluted with ultrapure water.

[0605] 7.2 Data Analysis

[0606] 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 using the following formula: tmin (%) = Peak area ratio tmin / peak area t0 ×100

[0607] The experimental results are shown in Table 7.

[0608] Table 7 Stability test results of representative compounds in human plasma

[0609] The results showed that the representative compounds 6-2, 11-2, and 17-2 of the present application had good stability in human plasma.

[0610] The above is an exemplary description of the implementation methods of the technical solution of the present invention. It should be understood that the scope of protection of the present invention is not limited to the above implementation methods. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of the present invention shall be included in the scope of protection of the claims of this application.

Claims

1. A compound of Formula I, its racemate, stereoisomer, tautomer, isotope-labeled form, solvate, polymorph, pharmaceutically acceptable salt, or prodrug thereof: in, R1, R2, R3 are the same or different and are independently selected from H, OH, CN, halogen, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 1-10 Alkoxy, halogenated C 1-10 Alkyl, halogenated C 1-10 Alkoxy, cyano C 1-10 Alkyl, cyano C 1-10 Alkoxy, C 3-10 Cycloalkyl; R4 is selected from H or R 41 Selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl-NH-, (C 1-6 Alkyl)2N-, C 1-6 Alkyl-NH-C 1-6 Alkyl, (C 1-6 alkyl)2N-C 1-6 Alkyl, C 1-6 Alkoxyalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-14 Aryl and 5-14 membered heteroaryl; R5 is selected from H, R 51 、R 52 The same or different, independently selected from H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl-NH-, (C 1-6 Alkyl)2N-, C 1-6 Alkyl-NH-C 1-6 Alkyl, (C 1-6 alkyl)2N-C 1-6 Alkyl, C 1-6 Alkoxyalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-14 Aryl and 5-14 membered heteroaryl; R 53 Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkyl-NH-, (C 1-6 Alkyl)2N-, C 1-6 Alkyl-NH-C 1-6 Alkyl, (C 1-6 alkyl)2N-C 1-6 Alkyl, C 1-6 Alkoxyalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-14 Aryl and 5-14 membered heteroaryl; Ring A is selected from C 3-8 Cycloalkyl, C 3-8 Heterocyclyl, Ra is selected from H, hydroxy, CN, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl; n is selected from 0, 1 or 2; q is selected from 0, 1 or 2; X is selected from CH or N; m is an integer selected from 0-6.

2. The compound according to claim 1, characterized in that R1 is selected from H, OH, CN, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl or halogenated C 1-6 alkoxy; Preferably, R1 is selected from H, OH, CN, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl or halo C 1-6 alkoxy; Preferably, R1 is selected from H, OH, Br, methyl, difluoromethoxy, 2,2,2-trifluoroethoxy, vinyl, cyclopropyl or ethynyl; Preferably, R1 is selected from H, OH, Br, methyl, difluoromethoxy, 2,2,2-trifluoroethoxy, vinyl or ethynyl; Preferably, R2 is selected from H, halogen, CN or C 1-6 alkyl; Preferably, R2 is selected from H or F; Preferably, R3 is selected from H or C 1-6 alkyl; Preferably, R3 is H.

3. The compound according to claim 1 or 2, characterized in that R4 is selected from H or For example Preferably, R4 is selected from H or Preferably, X-R4 is Preferably Preferably, X-R4 is -CH2-.

4. The compound according to any one of claims 1 to 3, characterized in that R5 is selected from H, Among them, R 51 is selected from H, methyl, ethyl, isopropyl or cyclopropyl; R 52 is selected from H or methyl; R 53 is selected from methyl; Ring A is selected from C 3-6 Cycloalkyl; Ra is selected from H, hydroxy, CN, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl; n is selected from 0 or 1; q is selected from 0 or 1; Preferably, ring A is selected from a cyclobutane ring; Preferably, R5 is selected from H, Preferably, R5 is selected from H, Preferably, R 51 is selected from H, methyl, ethyl, isopropyl or cyclopropyl; R 52 is selected from H or methyl; R 53 is selected from methyl; Ring A is selected from cyclobutane ring; Preferably, R5 is selected from H, 5. The compound according to any one of claims 1 to 4, characterized in that X is selected from CH or N; and when X is CH, R4 is H; or when X is N, R5 is H; Preferably, m is selected from 0, 1 or 2.

6. The compound according to any one of claims 1 to 5, characterized in that The structure of the compound shown in Formula I is shown below: wherein R1, R2, R4, R5, X, and m are independently defined as described in any one of claims 1 to 5; Preferably, the compound structure shown in Formula I is as follows: wherein R1, R2, and R5 are independently defined as defined in any one of claims 1 to 5; Preferably, the compound structure shown in Formula I is as follows: Among them, R1, R2, R 51 、R 52 , Ring A, Ra, m, n, q independently have the definitions as described in any one of claims 1 to 5.

7. The compound according to any one of claims 1 to 6, characterized in that The structure of the compound of formula I is shown below:

8. A structural fragment D having the structure of the compound of formula I according to any one of claims 1 to 7 after dehydrogenation, Preferably, the structure of D is as follows:

9. A compound represented by Formula V, its racemate, stereoisomer, tautomer, isotope-labeled form, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound: L'-D (Formula V) in, L' is a linker containing a linker portion M that can react with an antibody or an antigen-binding fragment thereof, and after L'-D reacts with the antibody or an antigen-binding fragment thereof, L' forms a linker L; Preferably, L' comprises a peptide residue L1 and a fragment L2 where the peptide residue is linked to D; D has the definition described in claim 8.

10. An antibody drug conjugate of formula VI, Ab-[LD] β (Formula VI) in, Ab is an antibody or an antigen-binding fragment thereof, D has the definition described in claim 8, L is a linker connecting Ab and D, and β is selected from an integer or decimal between 1 and 10.

11. A method for preparing the compound according to any one of claims 1 to 7, comprising the following scheme 1 or scheme 2: Option 1: includes the following steps: (1) Compound I-41 is deprotected by PG4 to obtain compound I-42; (2) Compound I-42 reacts with compound I-43 to obtain the compound of formula I; wherein R1, R2, R3, R4, R5, X, and m have the meanings as defined in any one of claims 1 to 7; Y is selected from a leaving group, such as OH, Cl, Br, and I; and PG4 is selected from an amino protecting group, such as Fmoc, Boc, Bn, and Cbz; Option 2: includes the following steps: (1) Compound I-51 is deprotected by PG5 to obtain compound I-52; (2) Compound I-52 reacts with compound I-53 to obtain the compound of formula I; Among them, R1, R2, R3, R4, R5, R 51 , X, m, and n have the definitions described in any one of claims 1 to 7; Y is selected from a leaving group, such as OH, Cl, Br, and I; and PG5 is selected from an amino protecting group, such as Fmoc, Boc, Bn, and Cbz.

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

13. Use of at least one of the compound according to any one of claims 1 to 7, the compound of formula V according to claim 9, or the antibody-drug conjugate of formula VI according to claim 10, their racemates, stereoisomers, tautomers, isotopically labeled forms, solvates, polymorphs, pharmaceutically acceptable salts, or prodrugs thereof, or the pharmaceutical composition according to claim 12 in the preparation of a topoisomerase I inhibitor and / or in the preparation of a medicament for preventing or treating a disease or condition associated with topoisomerase I; Preferably, the disease or condition is a tumor, and the tumor includes breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine 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 tumor disease, comprising administering to a patient a preventively 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, their racemates, stereoisomers, tautomers, isotope-labeled forms, solvates, polymorphs, pharmaceutically acceptable salts, or prodrug compounds thereof; Preferably, the tumor includes breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine 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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