WO2022166762A1 - 喜树碱类化合物及其制备方法和应用 - Google Patents

喜树碱类化合物及其制备方法和应用 Download PDF

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WO2022166762A1
WO2022166762A1 PCT/CN2022/074328 CN2022074328W WO2022166762A1 WO 2022166762 A1 WO2022166762 A1 WO 2022166762A1 CN 2022074328 W CN2022074328 W CN 2022074328W WO 2022166762 A1 WO2022166762 A1 WO 2022166762A1
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compound
hydrogen
membered
alkyl
formula
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PCT/CN2022/074328
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English (en)
French (fr)
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田强
张毅涛
缪羽
王波
叶健
汪小蓓
李德亮
李芬
宋宏梅
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四川科伦博泰生物医药股份有限公司
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Priority to CA3209426A priority Critical patent/CA3209426A1/en
Priority to CN202311518671.2A priority patent/CN117567478A/zh
Priority to AU2022216696A priority patent/AU2022216696A1/en
Priority to EP22749041.4A priority patent/EP4289851A1/en
Priority to KR1020237025527A priority patent/KR20230142710A/ko
Priority to JP2023544762A priority patent/JP2024506819A/ja
Priority to MX2023008716A priority patent/MX2023008716A/es
Priority to CN202280008418.XA priority patent/CN116829561A/zh
Publication of WO2022166762A1 publication Critical patent/WO2022166762A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/22—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains four or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/68037—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a camptothecin [CPT] or derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6843—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a material from animals or humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • This application is an application whose CN application number is 202110159956.6, the application date is February 5, 2021, the CN application number is 202110533304.4, and the application date is May 17, 2021, CN application number is 202110718245.8, and the application date is 2021
  • the present invention relates to a class of camptothecin compounds with antitumor activity and their conjugates, as well as their preparation methods and applications in the field of medicine.
  • Camptothecin (CPT, formula 1) is a pentacyclic quinoline nucleus compound isolated from the dove tree plant Camptotheca (Camptothecaacuminata). and ⁇ -hydroxylactone ring E, in which the 20-position is in the S configuration (see structural formula below). It was introduced into the clinic in the early 1970s due to its excellent anticancer activity, but later clinical trials were terminated due to severe side effects such as diarrhea and hemorrhagic cystitis.
  • camptothecin can form a ternary complex with cellular DNA topoisomerase I, thereby inhibiting the unwinding of DNA, leading to the blockage of DNA replication and resulting in cell death (Cancer Res. 1989, 49, 6365). Camptothecin and its derivatives have strong antitumor activity in animal in vivo models such as lung cancer, breast cancer, colorectal cancer, and ovarian cancer (Nature Review Cancer. 2006, 6, 789).
  • camptothecins have been approved and marketed 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 Exatecan, Rubitecan, Karenitecan, Diflomotecan, Lurtotecan, Gimatecan, Namitecan, Simmitecan, Silatecan, Chimmitecan, Elomotecan and the like.
  • Camptothecin drugs or derivatives often have hematological toxicity caused by bone marrow suppression, such as neutropenia, leukopenia, thrombocytopenia, anemia, etc., as well as gastrointestinal side effects, such as nausea, vomiting, diarrhea, etc.
  • Clinical studies have found that measures to improve the safety and efficacy of camptothecins include improving their pharmacokinetic properties, regulating their activity, reducing their dosage, or using their conjugates and antibodies to form antibody-drug conjugates. Therefore, there is still a high clinical demand and application value to develop camptothecin compounds and their conjugates with novel structures, which can improve efficacy and improve safety issues.
  • the present invention provides camptothecin compounds and their conjugates with novel structures.
  • the camptothecin compounds have good anti-tumor activity and are expected to be used for the treatment of tumor diseases; their conjugates are widely used in antibody conjugated drugs prospect.
  • a first aspect of the present invention provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotopic label, metabolite and prodrug thereof, wherein the compound has the following Structure shown:
  • R 1 and R 2 are each independently selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, hydroxy, cyano and C 3-6 cycloalkyl; or , R 1 and R 2 are connected with adjacent carbon atoms to form a 5-6 membered oxygen-containing heterocyclic ring;
  • R 3 is hydrogen or is connected to the ortho-position carbon atom of R 1 to form a six-membered carbocyclic ring;
  • A is selected from and one of the
  • R 4 is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl and 3-6 membered heterocyclyl;
  • R 5 and R 6 are each independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkylaminoalkyl, C 1-6 alkoxyalkyl, C 2-6 Alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, 3-6 membered heterocyclylalkyl, aryl and heteroaryl; or R 5 and R 6 with Adjacent carbon atoms are connected to form a 3-6 membered carbocyclic or heterocyclic ring;
  • the compound has the structure of formula (I):
  • R x is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl and 3-6 membered heterocycle base;
  • R y and R z are not both hydrogen and are independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylamine alkoxyalkyl, C 1-6 alkoxyalkyl, 3-6 membered heterocyclylalkyl and 3-6 membered heterocyclyl.
  • R x is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl, and 3-6 membered heterocyclyl ;
  • R y and R z are not both hydrogen and are independently selected from hydrogen, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkylaminoalkyl, C 1-6 alkoxyalkyl, 3-6 membered heterocyclylalkyl and 3-6 membered heterocyclyl.
  • R x is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl, 3 ⁇ 6-membered heterocyclyl;
  • R y and R z are not both hydrogen and are independently selected from hydrogen, C 2-6 alkenyl, C 2-6 alkynyl.
  • R x is selected from hydrogen or C 1-6 alkyl.
  • Rx is hydrogen
  • Ry and Rz are not both hydrogen and are independently selected from hydrogen, Dimethylaminomethylene, morpholinylmethylene and methoxymethylene.
  • Ry and Rz are not both hydrogen and are independently selected from hydrogen, Dimethylaminomethylene and methoxymethylene.
  • R y is hydrogen and R z is selected from Dimethylaminomethylene, morpholinylmethylene and methoxymethylene.
  • R y is hydrogen and R z is selected from Dimethylaminomethylene and methoxymethylene.
  • Rx is hydrogen
  • Ry is hydrogen
  • Rz is selected from Dimethylaminomethylene, morpholinylmethylene and methoxymethylene.
  • Rx is hydrogen
  • Ry is hydrogen
  • Rz is selected from and dimethylaminomethylene
  • the compound has the structure of formula (II):
  • A' is selected from and one of the
  • R x' is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl and 3-6 membered heterocyclyl;
  • R y' and R z' are independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 1-6 alkylaminoalkyl , C 3-6 Cycloalkyl, 3-6 membered heterocyclyl, 3-6 membered heterocyclylalkyl, C 2-6 alkenyl, C 2-6 alkynyl, aryl and heteroaryl, or R y' and R z ' is attached to adjacent carbon atoms into a 3-6 membered ring.
  • R x' is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl, 3-6 membered heteroalkyl ring base;
  • R y' and R z' are independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycle group, C 2-6 alkenyl, C 2-6 alkynyl, aryl, heteroaryl, or R y' and R z' are attached to adjacent carbon atoms to form a 3-6 membered ring.
  • the 3-6 membered ring is selected from a 3-6 membered carbocycle or a 3-6 membered heterocycle.
  • R x' is selected from hydrogen and C 1-6 alkyl.
  • Rx ' is selected from hydrogen and methyl.
  • R y' and R z' are independently selected from hydrogen, C 1-6 alkyl, C 1-6 alkoxyalkyl, C 1-6 alkylaminoalkyl, C 3-6 ring Alkyl and C 2-6 alkenyl, or R y' and R z' are attached to adjacent carbon atoms to form a 3-6 membered cycloalkyl.
  • R y' is selected from hydrogen and C 1-6 alkyl
  • R z' is selected from hydrogen, C 1-6 alkyl and C 3-6 cycloalkyl
  • R y' and R z' Linked to adjacent carbon atoms to form a 3-6 membered ring.
  • Ry' is selected from hydrogen and methyl
  • Rz' is selected from hydrogen, methyl, and cyclopropyl
  • Ry' and Rz' are attached to adjacent carbon atoms as a 3-membered carbocyclic ring.
  • Rx ' is selected from hydrogen and methyl
  • Ry' is selected from hydrogen and methyl
  • Rz' is selected from hydrogen, methyl and cyclopropyl
  • Adjacent carbon atoms are connected to form a 3-membered carbocyclic ring.
  • Rx ' is hydrogen
  • Ry' is selected from hydrogen and methyl
  • Rz' is selected from hydrogen, methyl, and cyclopropyl
  • Ry' and Rz' are attached to adjacent carbon atoms into a 3-membered carbon ring.
  • A' in formula (II) is
  • the compound has the structure of formula (III):
  • A" is selected from and one of the
  • R x " is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl and 3-6 membered heterocyclic group;
  • R y " and R z " are independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 1-6 alkylaminoalkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclyl, 3-6 membered heterocyclylalkyl, 4-6 membered heterocyclyl, C2-6 alkenyl, C2-6 alkynyl, aryl and heteroaryl , or R y " and R z " are connected to adjacent carbon atoms to form a 3-6 membered ring.
  • R x " is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl, 3-6 membered heteroalkyl ring base;
  • R y " and R z " are independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl, 4-6 membered heterocycle group, C 2-6 alkenyl, C 2-6 alkynyl, aryl, heteroaryl, or R y " and R z " are attached to adjacent carbon atoms to form a 3-6 membered ring.
  • R x " is selected from hydrogen and C 1-6 alkyl.
  • Rx" is hydrogen
  • the 3-6 membered ring is selected from a 3-6 membered carbocycle or a 3-6 membered heterocycle.
  • R y " and R z " are each independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 1-6 alkylamine alkyl, C 3-6 cycloalkyl and vinyl, or R y " and R z " are attached to adjacent carbon atoms to form a 3-6 membered ring.
  • R y " is hydrogen
  • R z " is selected from hydrogen, C 1-6 alkyl, C 3-6 cycloalkyl and vinyl, or R y " and R z " and adjacent carbon atoms Linked into a 3-6 membered carbocyclic ring.
  • Ry " is hydrogen
  • Rz " is selected from hydrogen, methyl, cyclopropyl, and vinyl
  • Ry " and Rz " are attached to adjacent carbon atoms to form a 3-membered carbocyclic ring.
  • Rx" is hydrogen
  • Ry " is hydrogen
  • Rz " is selected from hydrogen, methyl, cyclopropyl, and vinyl
  • Ry " and Rz " are attached to adjacent carbon atoms to form 3-membered carbon ring.
  • A" in formula (III) is
  • the compound has the structure of formula (IV):
  • R a and R b are independently selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 hydroxyalkyl, C 1-6 alkoxyalkyl, C 1-6 alkoxy, C 1-6 haloalkyl , hydroxyl and cyano groups; or R a and R b are connected with adjacent carbon atoms to form a 5-6 membered oxygen-containing heterocyclic ring;
  • R c and R d are independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 1-6 alkylaminoalkyl, C 3-6 cycloalkane base, 3-6 membered heterocyclyl, 3-6 membered heterocyclylalkyl, C2-6 alkenyl and C2-6 alkynyl, or R c and R d are attached to adjacent carbon atoms to form a 3-6 a membered carbocyclic or heterocyclic ring;
  • R e is selected from hydrogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl and C 2 -C 5 heterocyclyl;
  • R a and R b are independently selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, hydroxy, and cyano ; or R and R b is connected with adjacent carbon atoms to form a 5-6 membered oxygen-containing heterocyclic ring;
  • R c and R d are independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 1-6 alkylaminoalkyl, C 3-6 cycloalkane base, 3-6 membered heterocyclyl, 3-6 membered heterocyclylalkyl, C2-6 alkenyl and C2-6 alkynyl, or R c and R d are attached to adjacent carbon atoms to form a 3-6 a membered carbocyclic or heterocyclic ring;
  • R e is selected from hydrogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl and 4-6 membered heterocyclyl;
  • R a and R b are independently selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, hydroxy, and cyano or R a and R b are connected with adjacent carbon atoms to form a 5-6 membered oxygen-containing heterocyclic ring;
  • R c and R d are independently selected from hydrogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl , 4-6 membered heterocyclyl, C 2-6 alkenyl, C 2-6 alkynyl, or R c and R d are connected to adjacent carbon atoms to form a 3-6 membered carbocyclic or heterocyclic ring;
  • R e is selected from hydrogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl , or 4-6 membered heterocyclyl;
  • R a and R b are independently selected from hydrogen, halogen, and C 1-6 alkyl, or R a and R b are attached to adjacent carbon atoms to form a 5-6 membered oxygen-containing heterocycle.
  • R a and R b are independently selected from hydrogen, fluorine, chlorine and methyl, or R a and R b are jointly formed with the benzene ring to which they are attached
  • Z is selected from -CH2- , -CD2- , -CH2CH2- and -CF2- .
  • R a is methyl and R b is fluoro, or R a and R b are jointly formed with the benzene ring to which they are attached
  • Rc and Rd are independently selected from hydrogen, C 1-6 alkoxyalkyl and C 1-6 alkylaminoalkyl, or R c and R d are attached to adjacent carbon atoms to form a 3-6 membered carbocyclic ring.
  • R c is hydrogen and R d is selected from hydrogen, Methoxyethyl and cyclopropyl, or Rc and Rd are attached to adjacent carbon atoms to form a 3-6 membered carbocyclic ring.
  • R e is selected from hydrogen and C 1-6 alkyl.
  • R e is selected from hydrogen and isopropyl.
  • R a is methyl and R b is fluoro, or R a and R b are jointly formed with the benzene ring to which they are attached
  • R e is selected from hydrogen and isopropyl
  • R c is hydrogen
  • R d is selected from hydrogen, Methoxyethyl and cyclopropyl
  • Rc and Rd are attached to adjacent carbon atoms to form a 3-membered carbocyclic ring.
  • R a is methyl and R b is fluoro, or R a and R b are jointly formed with the benzene ring to which they are attached
  • R e is selected from hydrogen and isopropyl
  • R c is hydrogen
  • R d is selected from hydrogen, methoxyethyl and cyclopropyl
  • R c and R d are attached to adjacent carbon atoms to form a 3-membered carbocyclic ring.
  • the compound has the structure of formula (V):
  • R is selected from C 3-6 cycloalkyl and C 1-6 alkoxy
  • A"' is selected from and one of the
  • R x " ' is selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 3-6 cycloalkyl and 3-6 membered heterocyclyl;
  • R y " ' and R z " ' are independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 1-6 alkylaminoalkyl, C 3 -6 cycloalkyl, 3-6 membered heterocyclyl, 3-6 membered heterocyclylalkyl, C2-6 alkenyl, C2-6 alkynyl, aryl and heteroaryl, or R y " ' and R z " ' are connected to adjacent carbon atoms to form a 3-6 membered ring.
  • R is selected from methoxy and cyclopropyl.
  • the 3-6 membered ring is selected from a 3-6 membered carbocycle or a 3-6 membered heterocycle.
  • R y " ' and R z " ' are each independently selected from hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, C 1-6 Alkylaminoalkyl, C3-6cycloalkyl and vinyl, or Ry " ' and Rz " ' are attached to adjacent carbon atoms to form a 3-6 membered carbocyclic ring.
  • both Ry " ' and Rz " ' are hydrogen, or Ry " ' and Rz " ' are attached to adjacent carbon atoms to form a 3-6 membered carbocyclic ring.
  • R x " ' is selected from hydrogen and C 1-6 alkyl.
  • Rx" ' is hydrogen
  • A"' is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • R is selected from methoxy and cyclopropyl
  • Rx" ' is hydrogen
  • Ry " ' and Rz " ' are both hydrogen
  • Ry " ' and Rz " ' are adjacent to
  • the carbon atoms are linked into a 3-membered carbocyclic ring.
  • in formula (V)-1 as structure. In some embodiments, in formula (V)-1 as structure. In some embodiments, in formula (V)-1 as structure. In some embodiments, in formula (V)-1 as structure. In some embodiments, in formula (V)-1 as structure. In some embodiments, in formula (V)-1 as structure. In some embodiments, in formula (V)-1 as structure. In some embodiments, the present invention provides the following compounds:
  • the present invention also provides compounds of formula (VI) or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotopic labels, metabolites and prodrugs thereof:
  • M is the linker site with the antibody or its antigen-binding fragment
  • L is the linker connecting linker M and E;
  • E is the structural fragment connecting L and D;
  • D is the structural fragment of the cytotoxic drug.
  • M is selected from the following structures:
  • M is selected from the following structures:
  • L is selected from a divalent structure consisting of one or more of the following: C 1-6 alkylene, -N(R')-, carbonyl, -O-, Val, Cit, Phe, Lys, D-Val, Leu, Gly, Ala, Asn, Val-Cit, Val-Ala, Val-Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), D-Val-Leu- Lys, Gly-Gly-Arg, Ala-Ala-Asn, Ala-Ala-Ala, Val-Lys-Ala, Gly-Gly-Gly, Gly-Gly-Phe-Gly, Gly-Gly-Gly-Gly-Gly,
  • R' represents hydrogen, C 1-6 alkyl or -(CH 2 CH 2 O) r --containing alkyl; r is selected from an integer of 1-10; s is selected from an integer of 1-10.
  • L is selected from the following structures:
  • L is selected from the following structures:
  • E is selected from a single bond, -NH - CH2-,
  • E is -NH - CH2-.
  • the cytotoxic drug is selected from the -compounds of any one of the first aspects of the invention.
  • the cytotoxic drug is selected from compounds 1-1 to 1-15; 2-1 to 2-27; 3-1 to 3-26; 4-1 to 4-15 of the present invention ; or 5-1 to 5-36.
  • D is selected from the structure of a compound described herein after dehydrogenation.
  • D is selected from compounds 1-1 to 1-15; 2-1 to 2-27; 3-1 to 3-26; 4-1 to 4-15; or 5- 1 to 5-36 Structure after dehydrogenation.
  • D is selected from the following structures:
  • D is selected from the following structures:
  • M-L-E-D is selected from the following compounds:
  • M-L-E-D is selected from the following compounds:
  • an "*" marked in a compound structural formula indicates that the marked carbon atom is a chiral carbon atom, and the present invention includes a pair of enantiomers formed from this chiral carbon atom. If a compound contains two different chiral carbon atoms, the present invention includes 4 optical isomers formed by the chiral carbon atoms.
  • alkyl is defined as a straight or branched chain saturated aliphatic hydrocarbon group. In some embodiments, the alkyl group has 1 to 12, eg, 1 to 6, carbon atoms.
  • C 1-6 alkyl refers to a linear or branched group of 1 to 6 carbon atoms (eg, methyl, ethyl, n-propyl, isopropyl, n-butyl) , isobutyl, sec-butyl, tert-butyl, n-pentyl and n-hexyl), which are optionally substituted with 1 or more (such as 1, 2 or 3) suitable substituents.
  • alkenyl refers to a straight or branched chain hydrocarbon group containing at least one carbon-carbon double bond, including, for example, "C 2-6 alkenyl", “C 2-4 alkenyl” and the like. Examples include, but are not limited to: vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1,3-butadienyl, 1-pentenyl, 2-pentenyl Alkenyl, 3-pentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1,4-hexenyl Dienyl, etc.
  • alkynyl refers to a straight or branched chain hydrocarbon group containing at least one carbon-carbon triple bond. Included, for example, “C 2-6 alkynyl”, “C 4-6 alkynyl” and the like.
  • Examples include, but are not limited to: ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 1,3-butadiynyl, 1-pentynyl, 2 -Pentynyl, 3-pentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,4 -Hexadiynyl etc.
  • cycloalkyl refers to saturated cyclic hydrocarbon groups including, but not limited to, monocycloalkyl and bicycloalkyl groups (such as spirocycloalkyl, paracycloalkyl, and bridged cycloalkyl).
  • C 3-6 cycloalkyl refers to a cycloalkyl group having 3 to 6 ring carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc., which may be optionally replaced by 1 or Multiple (such as 1, 2 or 3) suitable substituents are substituted, eg methyl substituted cyclopropyl.
  • carrier refers to a saturated or partially unsaturated non-aromatic monocyclic or polycyclic structure, a hydrocarbon group attached through a ring carbon. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.
  • carrier refers to a saturated or unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (eg, a monocyclic ring such as a cyclopropane, cyclobutane, cyclopentane, cyclohexane, Cycloheptane, cyclooctane, cyclononane, or bicyclic rings, including spiro, fused, or bridged systems (such as bicyclo[1.1.1]pentane, bicyclo[2.2.1]heptane, Bicyclo[3.2.1]octane ring or bicyclo[5.2.0]nonane ring, decalin ring, etc.), which may be optionally substituted by 1 or more (such as 1, 2 or 3) suitable substituents Substituted.
  • 3-6 membered carbocycle refers to a carbocycle containing 3, 4, 5 or 6 ring-forming carbon atom
  • heterocyclyl refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic) non-aromatic cyclic structure whose ring atoms consist of carbon atoms and at least one (eg 1, 2 or 3) heteroatoms selected from nitrogen, oxygen and sulfur.
  • a heterocyclyl group can be attached to the rest of the molecule through any one of the ring atoms if the valence requirements are met.
  • the heterocyclic group in the present invention is preferably a 3- to 6-membered heterocyclic group.
  • 3-6 membered heterocyclic group refers to a heterocyclic group having 3 to 6 ring atoms, including 3-membered heterocyclic group, 4-membered heterocyclic group, 5-membered heterocyclic group and 6-membered heterocyclic group
  • the membered heterocyclic group includes nitrogen-containing heterocyclic group, oxygen-containing heterocyclic group, such as 4-6 membered heterocyclic group, such as 4-6 membered nitrogen-containing heterocyclic group, 4-6 membered oxygen-containing heterocyclic group.
  • heterocyclyl groups include, but are not limited to, azetidinyl, oxetanyl, tetrahydrofuryl, pyrrolidinyl, pyrrolidinonyl, Imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, piperazinyl, morpholinyl.
  • the heterocyclyl groups of the present invention may be optionally substituted with one or more substituents described herein.
  • Heterocyclyl groups in the present invention are optionally fused to one or more aromatic or non-aromatic rings.
  • oxygen-containing heterocycle refers to a heterocycle as previously described in which one or more (eg 1, 2 or 3) ring atoms are oxygen atoms, such as a 5-6 membered oxygen-containing heterocycle, specific examples include but Not limited to ethylene oxide ring, tetrahydrofuran ring, furan ring, tetrahydropyran ring, pyran ring and the like.
  • the "nitrogen-containing heterocycle” in the present invention refers to the aforementioned heterocycle in which one or more (eg 1, 2 or 3) ring atoms are nitrogen atoms.
  • haloalkyl refers to an alkyl group substituted with one or more (such as 1, 2 or 3) the same or different halogen atoms, wherein alkyl is as defined above.
  • C 1-6 haloalkyl refers to a haloalkyl having 1 to 6 carbon atoms.
  • Common haloalkyl groups include, but are not limited to, -CH2F , -CHF2 , -CF3 , -CH2CF3 , -CF2CF3 , -CH2CH2CF3 , -CH2Cl , and the like.
  • the haloalkyl groups of the present invention are optionally substituted with one or more substituents described herein.
  • aryl refers to a group obtained by removing a hydrogen atom from the carbon atom of the aromatic nucleus of an aromatic hydrocarbon molecule.
  • a 6-14 membered aryl group and specific examples include, but are not limited to, phenyl, naphthyl, anthracenyl, and the like.
  • heteroaryl refers to an aromatic cyclic group containing at least one ring member selected from N, O, and S. Specific examples include, but are not limited to, 5-6 membered heteroaryl, 5-6 membered nitrogen-containing heteroaryl, 5-6 membered oxygen-containing heteroaryl, and the like, such as furyl, thienyl, pyrrolyl, thiazolyl, isothiazole base, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1, 2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, pyrimidinyl, pyridazine group, pyrazinyl, 1,2,3-triazinyl,
  • alkoxy refers to a group having the structure "alkyl-O-", wherein alkyl is as defined above. For example, C 1-6 alkoxy, C 1-4 alkoxy, C 1-3 alkoxy or C 1-2 alkoxy and the like. Common alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy, hexyloxy Base et al.
  • the alkoxy groups in this invention are optionally substituted with one or more of the substituents described in this invention.
  • alkoxyalkyl refers to an alkyl group substituted with one or more (eg 1, 2, 3 or 4) alkoxy groups, wherein alkoxy and alkyl are as defined above .
  • C 1-6 alkoxyalkyl refers to 1-6 carbon atoms substituted with one or more (eg 1, 2, 3 or 4) alkoxy groups the alkyl group.
  • Common alkoxyalkyl groups include, but are not limited to, CH3O - CH2-, C2H5 - O - CH2- , C2H5 - O - CH2CH2- , and the like.
  • halo or halogen group is defined to include F, Cl, Br or I.
  • nitrogenide refers to an oxide (eg, a mono- or di-oxide) of at least one nitrogen atom in the structure of the compounds of the present application. Nitrogen mono-oxides can exist as single positional isomers or as mixtures of positional isomers.
  • substituted means that one or more (eg, one, two, three, or four) hydrogens on the designated atom are replaced by a selection from the designated group, provided that no more than the designated atom is present in the normal valences in the case and the substitutions form stable compounds. Combinations of substituents and/or variables are permissible only if such combinations form stable compounds.
  • substituent can be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more substituents from the list of substituents, one or more hydrogens on the carbon (to the extent of any hydrogens present) may be individually and/or together is replaced by an independently selected optional substituent. If a nitrogen of a substituent is described as being optionally substituted with one or more of the list of substituents, then one or more hydrogens on the nitrogen (to the extent of any hydrogens present) may each be independently selected optional substitution of substituents.
  • each substituent is selected independently of the other.
  • each substituent may be the same as or different from another (other) substituent.
  • one or more means 1 or more than 1, such as 2, 3, 4, 5 or 10, under reasonable conditions.
  • the point of attachment of a substituent can be from any suitable position on the substituent.
  • stereoisomer refers to isomers formed due to at least one asymmetric center. In compounds having one or more (eg, one, two, three or four) asymmetric centers, it may give rise to racemic mixtures, single enantiomers, diastereomeric mixtures and individual of diastereomers. Certain individual molecules can also exist as geometric isomers (cis/trans). Similarly, the compounds of the present invention may exist as mixtures of two or more structurally distinct forms in rapid equilibrium (often referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers Wait.
  • Solid line (—), solid wedge can be used in this paper or virtual wedge
  • the carbon-carbon bonds of the compounds of the present invention are depicted.
  • the use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers at that carbon atom are included (eg, a specific enantiomer, racemic mixture, etc.).
  • the use of real or dashed wedges to delineate bonds to asymmetric carbon atoms is intended to indicate that the indicated stereoisomer exists.
  • real and imaginary wedges are used to define relative, rather than absolute, stereochemistry.
  • the compounds of the present invention are intended to be available as stereoisomers (which include cis and trans isomers, optical isomers (eg, R and S enantiomers), diastereomers, Geometric isomers, rotational isomers, conformational isomers, atropisomers and mixtures thereof).
  • stereoisomers which include cis and trans isomers, optical isomers (eg, R and S enantiomers), diastereomers, Geometric isomers, rotational isomers, conformational isomers, atropisomers and mixtures thereof).
  • the compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (eg, racemic mixtures and pairs of diastereomers).
  • the present invention encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be a single polymorph or a mixture of more than one polymorph in any ratio.
  • compositions of the present invention may exist in free form for use in therapy, or, where appropriate, in the form of their pharmaceutically acceptable derivatives.
  • pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, metabolites or prodrugs, which can be directly or indirectly after administration to a patient in need thereof Compounds of the invention or metabolites or residues thereof are provided. Accordingly, references herein to "compounds of the present invention" are also intended to encompass the various derivative forms of the compounds described above.
  • Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.
  • Suitable acid addition salts are formed from acids that form pharmaceutically acceptable salts and include aspartate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, and the like.
  • Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts, including aluminum salts, arginine salts, choline salts, diethylamine salts, and the like.
  • esters means an ester derived from each of the compounds of the general formula in this application, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the invention in free acid or alcohol form).
  • physiologically hydrolyzable esters which can be hydrolyzed under physiological conditions to release the compounds of the invention in free acid or alcohol form.
  • the compounds of the present invention may themselves also be esters.
  • the compounds of the present invention may exist in the form of solvates, preferably hydrates, wherein the compounds of the present invention comprise a polar solvent as a structural element of the crystal lattice of the compound, in particular for example water, methanol or ethanol.
  • a polar solvent as a structural element of the crystal lattice of the compound, in particular for example water, methanol or ethanol.
  • the amount of polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratios.
  • metabolites of the compounds of the present invention ie substances formed in the body upon administration of the compounds of the present invention. Such products may result from, for example, oxidation, reduction, hydrolysis, amidation, deamidation, esterification, delipidation, enzymatic hydrolysis, and the like, of the administered compound. Accordingly, the present invention includes metabolites of the compounds of the present invention, including compounds prepared by methods of contacting a compound of the present invention with a mammal for a time sufficient to produce the metabolites thereof.
  • the present invention further includes within its scope prodrugs of the compounds of the present invention.
  • prodrugs will be functional derivatives of the compound that are readily converted in vivo to the desired therapeutically active compound.
  • the term "administration" as used in the methods of treatment of the present invention shall include the treatment of various diseases or conditions with prodrug forms of one or more of the claimed compounds, but in the The prodrug forms are converted in vivo to the compounds described above following administration to a subject.
  • “Design of Prodrug” ed. H. Bundgaard, Elsevier, 1985, conventional methods for selecting and preparing suitable prodrug derivatives are described.
  • the present invention further includes within its scope isotopic labels of the compounds of the present invention, which are identical to the compounds of the present invention except that one or more atoms have the same atomic number but an atomic mass or mass number different from that which predominates in nature Atomic substitution for atomic mass or mass number.
  • the present invention also encompasses compounds of the present invention that contain protecting groups.
  • protecting groups In any process for preparing the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any relevant molecule, thereby forming chemically protected forms of the compounds of the present invention. This can be accomplished by conventional protecting groups, such as those described in Protective Groups in Organic Chemistry, ed.J.F.W.McOmie, Plenum Press, 1973; and T.W.Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991 protecting groups, these references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of the first or second aspect of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvent thereof compound, nitrogen oxide, isotopic label, metabolite or prodrug, and one or more pharmaceutically acceptable carriers.
  • composition refers to a composition that can be used as a drug, comprising a pharmaceutically active ingredient (API) (or therapeutic agent) and optionally one or more pharmaceutically acceptable carriers.
  • pharmaceutically acceptable carrier refers to an excipient with which a therapeutic agent is administered and which, within the scope of sound medical judgment, is suitable for contact with human and/or other animal tissue without undue toxicity, irritation, Allergic reactions or other problems or complications with a reasonable benefit/risk ratio.
  • compositions described above can act systemically and/or locally, which can be achieved by suitable dosage forms.
  • dosage forms include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions , injectable solutions, elixirs, syrups.
  • compositions may contain 0.01 mg to 1000 mg of at least one compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotopic label, metabolite substances or prodrugs.
  • the present invention also provides a method for preparing the above-mentioned pharmaceutical composition or its corresponding formulation, which comprises combining at least one compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate thereof compound, nitrogen oxide, isotopic label, metabolite or prodrug in combination with one or more pharmaceutically acceptable carriers.
  • the present invention provides a kit product comprising:
  • kit product described above may contain 0.01 mg to 1000 mg of at least one compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotopic label, metabolite thereof substances or prodrugs.
  • the present invention also provides a method for preparing the above kit, which comprises adding at least one compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope thereof
  • a method for preparing the above kit comprises adding at least one compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope thereof
  • the marker, metabolite or prodrug or pharmaceutical composition described above is combined with the optional presence of at least one other therapeutic agent or a pharmaceutical composition, packaging and/or instructions comprising the other therapeutic agent.
  • the compounds of the present invention can exhibit strong effects on inhibiting abnormal cell proliferation.
  • the present application provides compounds of the invention or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotopic labels, metabolites and prodrugs or pharmaceutical combinations thereof of the invention It is used to treat diseases related to abnormal cell proliferation.
  • the present application also provides the compounds of the present invention or their pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotopic labels, metabolites and prodrugs or the above drugs Use of a composition in the manufacture of a medicament for the treatment of a disease in the aspect of abnormal cell proliferation.
  • the disorder of abnormal cell proliferation includes, but is not limited to, tumors, such as advanced solid tumors.
  • the application also provides compounds of the present invention or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotopic labels, metabolites and prodrugs thereof or pharmaceutical combinations of the present invention
  • a compound in the preparation of a preparation for inhibiting the proliferation of tumor cells is for in vivo or in vitro administration.
  • the formulation can be administered to a subject to inhibit the proliferation of tumor cells in the subject; alternatively, the formulation can be administered to cells in vitro (eg, cell lines or cells from the subject), to inhibit the proliferation of tumor cells in vitro.
  • the tumors described in the present invention include (but are not limited to): brain tumor, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer Cancer, colorectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, prostate cancer, female reproductive tract cancer, carcinoma in situ, lymphoma, neurofibromatosis, thyroid cancer, bone cancer, skin cancer, brain cancer cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, prostate tumor, mast cell tumor, multiple myeloma, melanoma, glioma or sarcoma.
  • the present invention provides a method for treating a disorder of abnormal cell proliferation, comprising the steps of: administering a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, stereoisomer thereof , polymorphs, solvates, nitrogen oxides, isotopic labels, metabolites and prodrugs or pharmaceutical compositions of the foregoing are administered to an individual in need thereof.
  • an effective amount refers to an amount sufficient to induce a biological or medical response in a cell, tissue, organ or organism (eg, an individual) and which is sufficient to achieve the desired prophylactic and/or therapeutic effect.
  • Dosage regimens can be adjusted to provide the optimal desired response. For example, it may be administered in a single dose, may be administered in divided doses over time, or may be administered in proportionately reduced or increased doses as appropriate. It will be appreciated that, for any particular individual, the specific dosing regimen should be adjusted as needed and according to the professional judgment of the person administering the composition or supervising the administration of the composition.
  • an effective amount is about 0.001-10000 mg/kg subject body weight/day. Where appropriate, an effective amount is about 0.01-1000 mg/kg subject body weight/day. About 0.01-1000 mg/kg of the subject's body weight can be administered daily, every two days, or every three days, usually about 0.1-500 mg/kg of the subject's body weight. Exemplary dosing regimens are one or more times per day, or one or more times per week, or one or more times per month.
  • the interval between single doses may generally be daily, weekly, monthly or yearly. Alternatively, it may be administered in a sustained release formulation, in which case less frequent dosing is required.
  • the dose and frequency of administration can vary depending on the half-life of the drug in the subject, as well as whether it is for prophylactic or therapeutic use. In prophylactic applications, relatively low doses are administered chronically at relatively infrequent intervals; in therapeutic applications, it is sometimes necessary to administer relatively high doses at shorter intervals until progression of the disease is delayed or stopped, preferably until the individual Shows partial or complete improvement in disease symptoms, after which prophylactic applications can be employed.
  • treating refers to alleviating or eliminating the targeted disease or condition. If a subject receives a therapeutic amount of a compound of the present invention, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the present invention, the subject exhibits at least one indicator and symptom that is observable and/or detectable A detected remission and/or improvement indicates that the subject has been successfully "treated". It is understood that treatment includes not only complete treatment, but also incomplete treatment, but the achievement of some biologically or medically relevant result.
  • administration/administrating/administration refers to the application of a pharmaceutically active ingredient (eg, a compound of the present invention) or a pharmaceutical composition comprising a pharmaceutically active ingredient (eg, a pharmaceutical composition of the present invention)
  • a pharmaceutically active ingredient eg, a compound of the present invention
  • a pharmaceutical composition comprising a pharmaceutically active ingredient (eg, a pharmaceutical composition of the present invention)
  • administration includes, but are not limited to, oral, subcutaneous, intramuscular, subperitoneal, ocular, nasal, sublingual, rectal, vaginal, and the like.
  • in need thereof refers to a physician or other caregiver's judgment that an individual needs or will benefit from a prophylactic and/or therapeutic procedure, based on the physician's or other caregiver's various areas of expertise. a factor.
  • the term "individual" refers to a human or non-human animal.
  • Individuals of the present invention include individuals (patients) with a disease and/or disorder and normal individuals.
  • the non-human animals of the present invention include all vertebrates, such as non-mammals, such as birds, amphibians, reptiles, etc., and mammals, such as non-human primates, livestock and/or domesticated animals (such as sheep, dogs, cats, cows, pigs, etc.).
  • the fourth aspect of the present invention provides a method for synthesizing the compound.
  • the compound of formula (I) in the present invention can be synthesized by the following synthetic route.
  • R x , R y and R z have the same meanings as described above;
  • LG is a leaving group selected from methanesulfonyl, trifluoromethanesulfonyloxy and halogen, preferably trifluoromethanesulfonyloxy or iodine;
  • the compound of formula (I)-IM1 is obtained by substitution reaction between the compound of formula (I)-SM1 and the compound of formula (I)-SM2.
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C;
  • this step is performed in a suitable organic solvent, which may be selected from halogenated hydrocarbons (eg, dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane ( 1,2-DCE), etc.), nitriles (such as acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylethyl acetate Amide (DMA), tetrahydrofuran (THF), 1,4-dioxane (Dioxane), dimethyl sulfoxide (DMSO) and any combination thereof, preferably acetonitrile.
  • halogenated hydrocarbons eg, dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane ( 1,2-DCE), etc.
  • nitriles such as acetonitrile (AN), etc.
  • NMP N,N-
  • this step is carried out in the presence of a suitable base, including an organic base or an inorganic base
  • the organic base may be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py)
  • the inorganic base may be selected from potassium phosphate (K 3 PO 4 ), sodium hydride (NaH), potassium carbonate (K 2 CO 3 ), carbonic acid sodium (Na 2 CO 3 ), sodium bicarbonate (NaHCO 3 ), cesium carbonate (Cs 2 CO 3 ) and NaOH, preferably Na 2 CO 3 or NaHCO 3 ;
  • the compound of formula (I) is obtained by condensation reaction between the compound of formula (I)-IM1 and the compound of formula (I)-SM3;
  • this step is performed under a suitable condensation reagent, which may be selected from HATU, HBTU, EDCI, DCC and HOBT, preferably HATU;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably DIPEA.
  • the compound of formula (II)-1 in the present invention can be synthesized and prepared by the following synthetic route.
  • R x' , R y' and R z' have the same meanings as described above;
  • LG is a leaving group selected from methanesulfonyl, trifluoromethanesulfonyloxy and halogen, preferably trifluoromethanesulfonyloxy or iodine;
  • PG is a protecting group, selected from
  • the compound of formula (II)-IM1 is obtained by a substitution reaction of the compound of formula (II)-SM1.
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 100°C, preferably 50°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably n-heptane.
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably n-heptane.
  • the compound of formula (II)-IM2 is obtained by reducing the compound of formula (II)-IM1;
  • this step is carried out in the presence of a suitable reducing agent, which can be selected from palladium catalysts, platinum catalysts, rhodium catalysts, preferably platinum catalysts;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 100°C, preferably 60°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate.
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate.
  • the compound of formula (II)-IM3 is obtained by the substitution reaction of the compound of formula (II)-IM2,
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 100°C, preferably 20°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate;
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate;
  • this step is performed under basic conditions, and reagents for providing basic conditions include organic bases and inorganic bases, including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide; the inorganic bases include but are not limited to potassium carbonate, sodium carbonate , sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, potassium hydroxide, preferably triethylamine.
  • organic bases and inorganic bases including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide
  • the inorganic bases include but are
  • the compound of formula (II)-IM4 is obtained by coupling reaction of the compound of formula (II)-IM3;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 70°C, 100°C, preferably 70°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, water, preferably a mixed solvent of tetrahydrofuran and water;
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, water, preferably a mixed solvent of tetrahydrofuran and water;
  • this step is performed under basic conditions, and reagents for providing basic conditions include organic bases and inorganic bases, including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide; the inorganic bases include but are not limited to potassium carbonate, sodium carbonate , sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, potassium hydroxide, preferably N,N-diisopropylethylamine.
  • organic bases and inorganic bases including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide;
  • the compound of formula (II)-IM5 is obtained by reducing the compound of formula (II)-IM4;
  • this step is carried out under a suitable reducing agent
  • the reducing agent can be selected from palladium catalyst, platinum catalyst, rhodium catalyst, preferably platinum catalyst;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 40°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably tetrahydrofuran.
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably tetrahydrofuran.
  • the compound of formula (II)-IM6 is obtained by ring-closing reaction of the compound of formula (II)-IM5;
  • this step is performed at a suitable temperature, said temperature being 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 5°C;
  • this step is performed in a suitable organic solvent selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, tert-butanol, preferably a mixed solvent of tetrahydrofuran and tert-butanol.
  • a suitable organic solvent selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, tert-butanol, preferably a mixed solvent of tetrahydrofuran and
  • the compound of formula (II)-IM7 is obtained by the substitution reaction of the compound of formula (II)-IM6;
  • this step is performed at a suitable temperature, said temperature being 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 5°C;
  • this step is performed in a suitable organic solvent selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably trifluoroacetic acid;
  • a suitable organic solvent selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably trifluoroacetic acid;
  • this step is performed under basic conditions, and reagents for providing basic conditions include organic bases and inorganic bases, including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide; the inorganic bases include but are not limited to potassium carbonate, sodium carbonate , sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, potassium hydroxide, preferably potassium tert-butoxide.
  • organic bases and inorganic bases including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide
  • the inorganic bases include
  • the compound of formula (II)-IM8 is obtained by reducing the compound of formula (II)-IM7;
  • this step is carried out under a suitable reducing agent
  • the reducing agent can be selected from palladium catalyst, platinum catalyst, rhodium catalyst, preferably palladium catalyst;
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 20°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably methanol.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably methanol.
  • the compound of formula (II)-IM9 is obtained by the substitution reaction of the compound of formula (II)-IM8;
  • this step is carried out at a suitable temperature, said temperature being 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 20°C;
  • this step is performed under basic conditions, and reagents for providing basic conditions include organic bases and inorganic bases, including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide; the inorganic bases include but are not limited to potassium carbonate, sodium carbonate , sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, potassium hydroxide, preferably pyridine.
  • organic bases and inorganic bases including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide
  • the inorganic bases include but are not limited to
  • the compound of formula (II)-IM10 is obtained by hydrolysis of the compound of formula (II)-IM9;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 60°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably methanol.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably methanol.
  • the reaction is carried out under acidic conditions, and reagents providing acidic conditions include hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably hydrochloric acid.
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 140°C;
  • this step is performed in a suitable organic solvent selected from the group consisting of toluene, methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably toluene.
  • a suitable organic solvent selected from the group consisting of toluene, methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably toluene.
  • the reaction is carried out under acidic conditions, and reagents providing acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably p-toluenesulfonic acid.
  • the compound of formula (II)-IM12 is obtained by hydrolysis of the compound of formula (II)-IM11;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 100°C;
  • the reaction is carried out under acidic conditions, and the reagents providing acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably hydrochloric acid.
  • the compound of formula (II)-IM13 is obtained by a substitution reaction between the compound of formula (II)-IM12 and the compound of formula (II)-SM2.
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C;
  • this step is performed in a suitable organic solvent, which may be selected from halogenated hydrocarbons (eg, dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane ( 1,2-DCE), etc.), nitriles (such as acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylethyl acetate Amide (DMA), tetrahydrofuran (THF), 1,4-dioxane (Diox), dimethyl sulfoxide (DMSO) and any combination thereof, preferably acetonitrile.
  • halogenated hydrocarbons eg, dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane ( 1,2-DCE), etc.
  • nitriles such as acetonitrile (AN), etc.
  • NMP N,N-di
  • this step is carried out in the presence of a suitable base, including an organic base or an inorganic base
  • the organic base may be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py)
  • the inorganic base may be selected from potassium phosphate (K 3 PO 4 ), sodium hydride (NaH), potassium carbonate (K 2 CO 3 ), Sodium carbonate (Na 2 CO 3 ), sodium bicarbonate (NaHCO 3 ), cesium carbonate (Cs 2 CO 3 ) and NaOH, preferably Na 2 CO 3 or NaHCO 3 ;
  • the compound of formula (II)-IM14 is obtained by condensation reaction between the compound of formula (II)-IM13 and the compound of formula (II)-SM3;
  • this step is performed under a suitable condensation reagent, which may be selected from HATU, HBTU, EDCI, DCC and HOBT, preferably HATU;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably DIPEA.
  • the compound of formula (II)-1 is obtained by acid hydrolysis of the compound of formula (II)-IM14;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and their mixed solvents, preferably a mixed solution of dichloromethane and methanol (volume ratio 2:1).
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and their mixed solvents, preferably a mixed solution of dichloromethane and methanol (volume ratio 2:1).
  • the reaction is carried out under acidic conditions, and the reagents providing acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably hydrochloric acid.
  • the compound of formula (II)-SM3-3 is obtained by substitution reaction between the compound of formula (II)-SM3-1 and the compound of formula (II)-SM3-2;
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 0-25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and mixed solvents thereof, preferably tetrahydrofuran.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and mixed solvents thereof, preferably tetrahydrofuran.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably K 2 CO 3 .
  • the compound of formula (II)-SM3 is obtained by hydrogenation of the compound of formula (II)-SM3-3;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and their mixed solvents, preferably methanol.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and their mixed solvents, preferably methanol.
  • this step is carried out under a suitable reducing agent
  • the reducing agent can be selected from palladium catalyst, platinum catalyst, rhodium catalyst, preferably palladium catalyst;
  • R y' and R z' are hydrogen:
  • the compound of formula (II)-SM3 is obtained by condensation reaction between the compound of formula (II)-SM3-4 and the compound of formula (II)-SM3-5;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and mixed solvents thereof, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and mixed solvents thereof, preferably N,N-dimethylformamide.
  • the compound of formula (II)-IM15 is obtained by condensation reaction between the compound of formula (II)-IM13 and the compound of formula (II)-SM4;
  • this step is performed under a suitable condensation reagent, which may be selected from HATU, HBTU, EDCI, DCC and HOBT, preferably HATU;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably DIPEA.
  • the compound of formula (II)-1 is obtained by removing the silicon protecting group from the compound of formula (II)-IM15;
  • the compound of formula (II) is obtained by condensation reaction between the compound of formula (II)-IM13 and the compound of formula (II)-SM5;
  • this step is performed under a suitable condensation reagent, which may be selected from the group consisting of HATU, HBTU, EDCI, DCC and HOBT, preferably HATU;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably DIPEA.
  • the compound of formula (III)-1 in the present invention can be synthesized and prepared by the following synthetic route.
  • R x ", R y “ and R z " have the same meanings as described above;
  • LG is a leaving group selected from methanesulfonyl, trifluoromethanesulfonyloxy and halogen, preferably trifluoromethanesulfonyloxy or chlorine;
  • PG is a protecting group, selected from
  • the compound of formula (III)-IM1 is obtained by a substitution reaction of the compound of formula (III)-SM1.
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 100°C, preferably 50°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably n-heptane.
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably n-heptane.
  • the compound of formula (III)-IM2 is obtained by reducing the compound of formula (III)-IM1;
  • this step is carried out in the presence of a suitable reducing agent, which can be selected from palladium catalysts, platinum catalysts, rhodium catalysts, preferably platinum catalysts;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 100°C, preferably 60°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate.
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate.
  • the compound of formula (III)-IM3 is obtained by the substitution reaction of the compound of formula (III)-IM2,
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 100°C, preferably 20°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate;
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate;
  • this step is performed under basic conditions, and reagents for providing basic conditions include organic bases and inorganic bases, including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide; the inorganic bases include but are not limited to potassium carbonate, sodium carbonate , sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, potassium hydroxide, preferably triethylamine.
  • organic bases and inorganic bases including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide
  • the inorganic bases include but are
  • the compound of formula (III)-IM4 is obtained by coupling reaction of the compound of formula (III)-IM3;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 70°C, 100°C, preferably 70°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, water, preferably a mixed solvent of tetrahydrofuran and water;
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, water, preferably a mixed solvent of tetrahydrofuran and water;
  • this step is performed under basic conditions, and reagents for providing basic conditions include organic bases and inorganic bases, including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide; the inorganic bases include but are not limited to potassium carbonate, sodium carbonate , sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, potassium hydroxide, preferably N,N-diisopropylethylamine.
  • organic bases and inorganic bases including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide;
  • the compound of formula (III)-IM5 is obtained by reduction reaction of the compound of formula (III)-IM4;
  • this step is carried out under a suitable reducing agent
  • the reducing agent can be selected from palladium catalyst, platinum catalyst, rhodium catalyst, preferably platinum catalyst;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 40°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably tetrahydrofuran.
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably tetrahydrofuran.
  • the compound of formula (III)-IM6 is obtained by ring-closing reaction of the compound of formula (III)-IM5;
  • this step is performed at a suitable temperature, said temperature being 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 5°C;
  • this step is performed in a suitable organic solvent selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, tert-butanol, preferably a mixed solvent of tetrahydrofuran and tert-butanol.
  • a suitable organic solvent selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, tert-butanol, preferably a mixed solvent of tetrahydrofuran and
  • the compound of formula (III)-IM7 is obtained by the substitution reaction of the compound of formula (III)-IM6;
  • this step is performed at a suitable temperature, said temperature being 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 5°C;
  • this step is performed in a suitable organic solvent selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably trifluoroacetic acid;
  • a suitable organic solvent selected from the group consisting of trifluoroacetic acid, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably trifluoroacetic acid;
  • this step is performed under basic conditions, and reagents for providing basic conditions include organic bases and inorganic bases, including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide; the inorganic bases include but are not limited to potassium carbonate, sodium carbonate , sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, potassium hydroxide, preferably potassium tert-butoxide.
  • organic bases and inorganic bases including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide
  • the inorganic bases include
  • the compound of formula (III)-IM8 is obtained by reducing the compound of formula (III)-IM7;
  • this step is carried out under a suitable reducing agent
  • the reducing agent can be selected from palladium catalyst, platinum catalyst, rhodium catalyst, preferably palladium catalyst;
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 20°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably methanol.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably methanol.
  • the compound of formula (III)-IM9 is obtained by the substitution reaction of the compound of formula (III)-IM8;
  • this step is carried out at a suitable temperature, said temperature being 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 20°C;
  • this step is performed under basic conditions, and reagents for providing basic conditions include organic bases and inorganic bases, including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide; the inorganic bases include but are not limited to potassium carbonate, sodium carbonate , sodium bicarbonate, potassium tert-butoxide, sodium hydride, sodium hydroxide, potassium hydroxide, preferably pyridine.
  • organic bases and inorganic bases including but not limited to triethylamine, pyridine, N,N-diisopropylamine Propylethylamine, n-butyllithium, lithium diisopropylamide, lithium bis-trimethylsilyl amide, sodium bis-trimethylsilyl amide
  • the inorganic bases include but are not limited to
  • the compound of formula (III)-IM10 is obtained by hydrolysis of the compound of formula (III)-IM9;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 60°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably methanol.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably methanol.
  • the reaction is carried out under acidic conditions, and reagents providing acidic conditions include hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably hydrochloric acid.
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 140°C;
  • this step is performed in a suitable organic solvent selected from the group consisting of toluene, methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably toluene.
  • a suitable organic solvent selected from the group consisting of toluene, methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dichloromethane Methyl sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably toluene.
  • the reaction is carried out under acidic conditions, and reagents providing acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably p-toluenesulfonic acid.
  • the compound of formula (III)-IM12 is obtained by hydrolysis of the compound of formula (III)-IM11;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 100°C;
  • the reaction is carried out under acidic conditions, and the reagents providing acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably hydrochloric acid.
  • the compound of formula (III)-IM13 is obtained by a substitution reaction between the compound of formula (III)-IM12 and the compound of formula (III)-SM2.
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C;
  • this step is performed in a suitable organic solvent, which may be selected from halogenated hydrocarbons (eg, dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane ( 1,2-DCE), etc.), nitriles (such as acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylethyl acetate Amide (DMA), tetrahydrofuran (THF), 1,4-dioxane (Dioxane), dimethyl sulfoxide (DMSO) and any combination thereof, preferably acetonitrile.
  • halogenated hydrocarbons eg, dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane ( 1,2-DCE), etc.
  • nitriles such as acetonitrile (AN), etc.
  • NMP N,N-
  • this step is carried out in the presence of a suitable base, including an organic base or an inorganic base
  • the organic base may be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py)
  • the inorganic base may be selected from potassium phosphate (K 3 PO 4 ), sodium hydride (NaH), potassium carbonate (K 2 CO 3 ), Sodium carbonate (Na 2 CO 3 ), sodium bicarbonate (NaHCO 3 ), cesium carbonate (Cs 2 CO 3 ) and NaOH, preferably Na 2 CO 3 or NaHCO 3 ;
  • the compound of formula (III)-IM14 is obtained by condensation reaction between the compound of formula (III)-IM13 and the compound of formula (III)-SM3;
  • this step is performed under a suitable condensation reagent, which may be selected from HATU, HBTU, EDCI, DCC and HOBT, preferably HATU;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is performed in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably DIPEA.
  • the compound of formula (III)-1 is obtained by acid hydrolysis reaction of the compound of formula (III)-IM14;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and their mixed solvents, preferably a mixed solution of dichloromethane and methanol (volume ratio 2:1).
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and their mixed solvents, preferably a mixed solution of dichloromethane and methanol (volume ratio 2:1).
  • the reaction is carried out under acidic conditions, and the reagents providing acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably hydrochloric acid.
  • the compound of formula (III)-SM3-3 is obtained by substitution reaction between the compound of formula (III)-SM3-1 and the compound of formula (II)-SM3-2;
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 0-25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and mixed solvents thereof, preferably tetrahydrofuran.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and mixed solvents thereof, preferably tetrahydrofuran.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably K 2 CO 3 .
  • the compound of formula (III)-SM3 is obtained by hydrogenation of the compound of formula (III)-SM3-3;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and their mixed solvents, preferably methanol.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and their mixed solvents, preferably methanol.
  • this step is carried out under a suitable reducing agent
  • the reducing agent can be selected from palladium catalyst, platinum catalyst, rhodium catalyst, preferably palladium catalyst;
  • R y' and R z' are hydrogen:
  • the compound of formula (III)-SM3 is obtained by condensation reaction between the compound of formula (III)-SM3-4 and the compound of formula (III)-SM3-5;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and mixed solvents thereof, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate and mixed solvents thereof, preferably N,N-dimethylformamide.
  • the compound of formula (III)-IM15 is obtained by condensation reaction between the compound of formula (III)-IM13 and the compound of formula (III)-SM4;
  • this step is performed under a suitable condensation reagent, which may be selected from HATU, HBTU, EDCI, DCC and HOBT, preferably HATU;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably DIPEA.
  • the compound of formula (III)-1 is obtained by removing the silicon protecting group from the compound of formula (III)-IM15;
  • the compound of formula (III)-1 is obtained by condensation reaction between the compound of formula (III)-IM13 and the compound of formula (III)-SM5;
  • this step is performed under a suitable condensation reagent, which may be selected from HATU, HBTU, EDCI, DCC and HOBT, preferably HATU;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably DIPEA.
  • the compound of formula (IV) in the present invention can be synthesized and prepared by the following synthetic route:
  • R a , R b , R c , R d and R e have the same meanings as described above;
  • the compound of formula (IV)-IM1 is obtained by the nitration reaction of the compound of formula (IV)-SM1;
  • this step is carried out at a suitable temperature, said temperature being 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.
  • the compound of formula (IV)-IM2 is obtained by hydrogenation of the compound of formula (IV)-IM1;
  • this step is carried out under a suitable reducing agent
  • the reducing agent can be selected from palladium catalyst, platinum catalyst, rhodium catalyst, preferably palladium catalyst;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate.
  • the compound of formula (IV)-IM3 is obtained by acylation of the compound of formula (IV)-IM2,
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 100°C, preferably 25°C;
  • the compound of formula (IV)-IM4 is obtained by reacting the compound of formula (IV)-IM3 with DMF-DMA;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 120°C;
  • the compound of formula (IV)-IM5 is obtained by substitution reaction of the compound of formula (IV)-IM4 and formula (IV)-SM2;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C;
  • this step is carried out in a suitable organic solvent, which can be selected from methanol, ethanol, N,N-methylpyrrolidone, dimethyl sulfoxide, preferably ethanol;
  • the compound of formula (IV)-IM6 is obtained by reduction reaction of the compound of formula (IV)-IM5;
  • this step is carried out under a suitable reducing agent, preferably sodium borohydride;
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 0-25°C;
  • this step is carried out in a suitable organic solvent, and the organic solvent can be selected from tetrahydrofuran, glacial acetic acid, methanol and mixed solutions thereof, preferably glacial acetic acid.
  • the organic solvent can be selected from tetrahydrofuran, glacial acetic acid, methanol and mixed solutions thereof, preferably glacial acetic acid.
  • the compound of formula (IV)-IM7 is obtained by protecting the amino group of the compound of formula (IV)-IM6 with Fmoc;
  • the compound of formula (IV)-IM9 is obtained by ring-closure reaction of the compound of formula (IV)-IM8 under acidic conditions;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 120°C;
  • this step is performed in a suitable organic solvent selected from the group consisting of toluene, xylene, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide, toluene and xylene are preferred;
  • this step is performed under acidic conditions
  • Reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably p-toluenesulfonic acid.
  • the Fmoc protecting group is removed by the compound of formula (IV)-IM9 to obtain the compound of formula (IV)-IM10;
  • the compound of formula (IV) is obtained by condensation reaction between the compound of formula (IV)-IM10 and the compound of formula (IV)-SM4;
  • this step is performed under a suitable condensation reagent, which may be selected from HATU, HBTU, EDCI, DCC and HOBT, preferably HBTU;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably DIPEA.
  • LG is a leaving group selected from methanesulfonyl, trifluoromethanesulfonyloxy and halogen, preferably trifluoromethanesulfonyloxy or iodine;
  • the compound of formula (IV)-IM11 is obtained by reducing the compound of formula (IV)-SM5,
  • this step is carried out in the presence of a suitable reducing agent, which can be selected from palladium catalysts, platinum catalysts, rhodium catalysts, preferably platinum catalysts;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate and tetrahydrofuran.
  • a suitable organic solvent selected from the group consisting of tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide , n-heptane, n-hexane, ethyl acetate, preferably ethyl acetate and tetrahydrofuran.
  • the compound of formula (IV)-IM12 is obtained by Friedel-Crafts acylation of the compound of formula (IV)-IM11;
  • this step is carried out at a suitable temperature, said temperature being 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.
  • the compound of formula (IV)-IM13 is obtained by ring-closure reaction of formula (IV)-IM12 under acidic conditions;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 120°C;
  • this step is performed in a suitable organic solvent selected from the group consisting of toluene, xylene, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide, toluene and xylene are preferred;
  • this step is performed under acidic conditions
  • Reagents that provide acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably p-toluenesulfonic acid.
  • the compound of formula (IV)-IM14 is obtained by substitution reaction of formula (IV)-IM13;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent, which can be selected from methanol, ethanol, N,N-methylpyrrolidone, dimethyl sulfoxide, preferably dimethyl sulfoxide;
  • the compound of formula (IV)-IM15 is obtained by reduction reaction of the compound of formula (IV)-IM14,
  • this step is carried out in the presence of a suitable reducing agent, which may be selected from the group consisting of palladium catalysts, platinum catalysts, rhodium catalysts, triphenylphosphine, triethyl phosphite, preferably triethyl phosphite ;
  • a suitable reducing agent which may be selected from the group consisting of palladium catalysts, platinum catalysts, rhodium catalysts, triphenylphosphine, triethyl phosphite, preferably triethyl phosphite ;
  • this step is carried out at a suitable temperature, said temperature being 20°C, 25°C, 50°C, 60°C, 80°C, 100°C, preferably 80°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, toluene, and mixed solutions thereof, preferably a mixed solution of methanol and toluene.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, toluene, and mixed solutions thereof, preferably a mixed solution of methanol and toluene.
  • the compound of formula (IV)-IM16 is obtained by substitution reaction between the compound of formula (IV)-IM15 and the compound of formula (IV)-SM6.
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 50°C;
  • this step is performed in a suitable organic solvent, which may be selected from halogenated hydrocarbons (eg, dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane ( 1,2-DCE), etc.), nitriles (such as acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylethyl acetate Amide (DMA), tetrahydrofuran (THF), 1,4-dioxane (Dioxane), dimethyl sulfoxide (DMSO) and any combination thereof, preferably acetonitrile.
  • halogenated hydrocarbons eg, dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane ( 1,2-DCE), etc.
  • nitriles such as acetonitrile (AN), etc.
  • NMP N,N-
  • this step is carried out in the presence of a suitable base, including an organic base or an inorganic base
  • the organic base may be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py)
  • the inorganic base may be selected from potassium phosphate (K 3 PO 4 ), sodium hydride (NaH), potassium carbonate (K 2 CO 3 ), carbonic acid sodium (Na 2 CO 3 ), sodium bicarbonate (NaHCO 3 ), cesium carbonate (Cs 2 CO 3 ) and NaOH, preferably Na 2 CO 3 or NaHCO 3 ;
  • the compound of formula (IV) is obtained by condensation reaction between the compound of formula (IV)-IM16 and the compound of formula (IV)-SM4;
  • this step is carried out with a suitable condensing reagent, which can be selected from HATU, HBTU, EDCI, DCC and HOBT, preferably HBTU;
  • this step is performed at a suitable temperature, said temperature being 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C;
  • this step is carried out in a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • a suitable organic solvent selected from methanol, tetrahydrofuran, dichloromethane, N,N-dimethylformamide, N-methylpyrrolidone, dimethylformamide Sulfoxide, n-heptane, n-hexane, ethyl acetate, preferably N,N-dimethylformamide.
  • this step is performed in a suitable base, including an organic base or an inorganic base
  • the organic base can be selected from DIPEA, TEA, t-BuOK and Py
  • the inorganic base can be selected from K 3 PO 4 , NaH, K 2 CO 3 , Na 2 CO 3 , Cs 2 CO 3 and NaOH, preferably DIPEA.
  • the compound of formula (V)-1 in the present invention can use starting materials Synthetic preparation is carried out according to the same synthetic route of formula (III).
  • the present invention provides camptothecin compounds represented by formula (I) to formula (IV), as well as pharmaceutical compositions, preparation methods and uses thereof. Such compounds have good antitumor activity, have the potential to overcome drug resistance, and can be used to treat abnormal cell proliferation disorders, including but not limited to advanced solid tumors.
  • the measuring instrument of nuclear magnetic resonance used a Bruker 400 MHz nuclear magnetic resonance apparatus; hexadeuterated dimethyl sulfoxide (DMSO-d 6 ); and the internal standard substance was tetramethylsilane (TMS).
  • NMR nuclear magnetic resonance
  • s singlet
  • d doublet
  • t triplet
  • q quartet
  • m multiplet
  • br broad
  • J coupling constant
  • Hz Hertz
  • DMSO-d6 deuterated dimethyl sulfoxide.
  • the delta value is expressed in ppm value.
  • the measuring instrument of mass spectrometry used Agilent (ESI) mass spectrometer, model Agilent 6120B.
  • the compound 2-hydroxypent-3-ynoic acid (4.29 mg, 37.63 ⁇ mol) was dissolved in DMF (1 mL), HATU (21.46 mg, 56.44 ⁇ mol), SM1-1 (10.00 mg, 22.94 ⁇ mol) and DIPEA (7.29 mg) were added , 56.44 ⁇ mol), 25 °C reaction for 2 hours.
  • the reaction solution was concentrated under reduced pressure, and the concentrate was directly purified by preparative high performance liquid chromatography (conditions are as follows) to obtain the title compounds 1-1-A 3.24 mg and 1-1-B 3.98 mg.
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the characterization data of 1-1-A structure are as follows:
  • the characterization data of the 1-1-B structure are as follows:
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the characterization data of 1-7-A structure are as follows:
  • the characterization data of the 1-7-B structure are as follows:
  • compound 2-1-01 (5.00 g, 29.14 mmol) was dissolved in n-heptane (25 mL), concentrated sulfuric acid (25 mL) was added, heated to 50 °C, and NBS (6.22 mL) was added in batches at 50 °C.
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • compound 2-1-03 (3.63 g, 14.82 mmol) was dissolved in ethyl acetate (70 mL), and triethylamine (4.50 g, 44.45 mmol) and acetic anhydride (2.27 g, 22.23 mmol) were added to keep the solution.
  • the reaction was carried out at 20°C for 20 hours, and the reaction was monitored by high performance liquid chromatography coupled with mass spectrometry. Water was added to the reaction solution, extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product.
  • Step seven (Z)-N-(3-chloro-7-(hydroxyimino)-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide
  • potassium tert-butoxide (1.50 g, 13.37 mmol) was dissolved in THF (16 mL) and tert-butanol (4 mL), and compound 2-1-07 (1.53 g, 6.08 mmol) in THF was added dropwise.
  • amyl nitrite (1.14 g, 9.73 mmol) was added dropwise to the solution (16 mL), and the reaction was kept at 5°C for 1 hour.
  • the reaction was monitored by high performance liquid chromatography coupled with mass spectrometry.
  • Step 8 N-(7-Amino-3-chloro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide
  • Step 9 Synthesis of N,N'-(3-chloro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1,7-diyl)diacetamide
  • Step 10 Synthesis of N-(8-amino-6-chloro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)acetamide
  • Step eleven N-((9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexa Synthesis of Hydro-1H,12H-benzopyrano[3',4':6,7]indolazino[1,2-b]quinolin-1-yl)acetamide
  • Step twelve (9S)-1-amino-5-chloro-9-ethyl-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzene Synthesis of Pyrano[3',4':6,7]indolazino[1,2-b]quinoline-10,13-dione
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% trifluoroacetic acid)
  • the structural representation data is as follows:
  • the trifluoroacetate salt of compound 2-23 (40.00 mg, 81.91 ⁇ mol) was dissolved in N,N-dimethylformamide (1 mL), followed by the addition of 2-((tert-butyldiphenyl Silyl)oxy)acetic acid (30.91mg, 98.29 ⁇ mol), HATU (62.25mg, 163.81 ⁇ mol) and N,N-diisopropylethylamine (42.34mg, 327.63 ⁇ mol), kept at 25°C for 0.5 hour reaction, with The reaction was monitored by high performance liquid chromatography coupled with chromatography.
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the characterization data of 2-1-B structure are as follows:
  • Step 1 (2S)-2-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl -10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizine Iso[1,2-b]quinolin-1-yl)propylamine and (2S)-2-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)-5-chloro -9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano Synthesis of [3',4':6,7]indolazino[1,2-b]quinolin-1-yl)propy
  • the hydrochloride of 2-23 (30.00 mg, 61.43 ⁇ mol) was dissolved in N,N-dimethylformamide (1 mL), followed by adding (S)-2-((tert-butyldiphenyl) Silyl)oxy)propionic acid (24.21mg, 73.72 ⁇ mol), HATU (35.01mg, 92.14 ⁇ mol) and N,N-diisopropylethylamine (23.82mg, 184.29 ⁇ mol), kept at 25°C for 1 hour, The reaction was monitored by high performance liquid chromatography coupled with mass spectrometry.
  • Step 2 (2S)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10, 13,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolino[1,2-b]quinolin-1-yl)-2 -Hydroxypropylamine and (2S)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10 ,13,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)- Synthesis of 2-hydroxypropylamine
  • 2-7-01-A (6.00 mg, 7.87 ⁇ mol) and 2-7-01-B (6.00 mg, 7.87 ⁇ mol) were dissolved in anhydrous tetrahydrofuran (1 mL) in two reaction flasks at 25°C, respectively.
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the characterization data of 2-7-A structure are as follows:
  • the characterization data of 2-7-B structure are as follows:
  • Step 1 (2S)-2-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl- 10,13-Dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino [1,2-b]Quinolin-1-yl)-2-cyclopropylacetamide and (2S)-2-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S )-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[ de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-2-cycloprop
  • the hydrochloride of 2-23 (30.00 mg, 61.43 ⁇ mol) was dissolved in N,N-dimethylformamide (1 mL), followed by the addition of 2-((tert-butyldiphenylsilyl) oxy)-2-cyclopropylacetic acid (26.13mg, 73.72 ⁇ mol), HATU (35.01mg, 92.14 ⁇ mol) and N,N-diisopropylethylamine (23.82mg, 184.29 ⁇ mol), kept at 25°C for reaction 1 hours, the reaction was monitored by high performance liquid chromatography coupled to mass spectrometry.
  • Step 2 (2S)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10, 13,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolino[1,2-b]quinolin-1-yl)-2 -Cyclopropyl-2-hydroxyacetamide and (2S)-N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo- 2,3,9,10,13,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolazino[1,2-b]quino Lin-1-yl)-2-cyclopropyl-2-hydroxyacetamide and (2R)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy
  • 2-12-01-A (8.00 mg, 10.15 ⁇ mol) and 2-12-01-B (10.00 mg, 12.68 ⁇ mol) were dissolved in anhydrous tetrahydrofuran (1 mL) in two reaction flasks at 25°C, respectively.
  • reaction solution was purified by preparative high performance liquid chromatography, two isomer products were separated from the reaction using 2-12-01-A as the raw material, and the preparation solution was lyophilized to obtain compound 2-12-A (0.77 mg), 2-12-B (1.03 mg); 2-12-01-B was used as the raw material to separate two isomer products, and the prepared liquids were lyophilized to obtain compound 2-12-C (2.50 mg) , 2-12-D (1.00 mg).
  • 2-12-A/2-12-B purification conditions are as follows:
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the peak retention time is: 2-12-A: 10.0-11.0min, 2-12-B: 11.0-12.5min
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the peak retention time is: 2-12-C: 10.6-11.4min, 2-12-D: 11.4-12.5min
  • the characterization data of 2-12-A structure are as follows:
  • the characterization data of 2-12-B structure are as follows:
  • the characterization data of 2-12-C structure are as follows:
  • the 2-12-D structural characterization data are as follows:
  • the hydrochloride of 2-23 (30.00 mg, 61.43 ⁇ mol) was dissolved in N,N-dimethylformamide (1 mL), followed by the addition of 2-((tert-butyldimethylsilyl) oxy)-2-methylpropionic acid (16.10 mg, 73.72 ⁇ mol), HATU (35.01 mg, 92.14 ⁇ mol) and N,N-diisopropylethylamine (23.82 mg, 184.29 ⁇ mol), kept at 25°C for reaction 1 hours, the reaction was monitored by high performance liquid chromatography coupled to mass spectrometry.
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the peak retention times are: 2-17-A: 9.5-10.2min and 2-17-B: 10.4-10.6min.
  • Step 1 1-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13 -Dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolazino[1, 2-b]Quinolin-1-yl)cyclopropane-1-carboxamide and 1-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)-5-chloro-9 -Ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3 Synthesis of ',4':6,7]indolazino[1,2-b]quinolin-1-yl)cyclopropane
  • the hydrochloride of 2-23 (30.00 mg, 61.43 umol) was dissolved in N,N-dimethylformamide (1 mL), followed by adding 1-((tert-butyldiphenylsilyl) Oxy)cyclopropane-1-carboxylic acid (25.10mg, 73.72umol), HATU (35.01mg, 92.14umol) and N,N-diisopropylethylamine (23.82mg, 184.29umol), keep 25 °C for reaction 1 hours, the reaction was monitored by high performance liquid chromatography coupled to mass spectrometry.
  • Step 2 N-((1S,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15- Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)-1-hydroxycyclopropane -1-Carboxamide and N-((1R,9S)-5-chloro-9-ethyl-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13 ,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolino[1,2-b]quinolin-1-yl)-1- Synthesis of Hydroxycyclopropane-1-carboxamide
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the characterization data of 2-20-A structure are as follows:
  • the characterization data of 2-20-B structure are as follows:
  • the structural representation data is as follows:
  • the structural representation data is as follows:
  • the structural representation data is as follows:
  • the structural representation data is as follows:
  • reaction solution was slowly poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, then filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product, which was purified by flash silica gel column , to obtain the title compound 0.43g.
  • the structural representation data is as follows:
  • Step 6 Synthesis of N-(4-chloro-3-fluoro-7-(hydroxyimino)-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide
  • Tetrahydrofuran (16 mL) and tert-butanol (4 mL) were added to the reaction flask, and after cooling to 5°C in an ice bath, potassium tert-butoxide (415.18 mg, 3.70 mmol) was added, and then compound 3-1-06 (0.43 mg) was added.
  • 1.68 mmol was dissolved in tetrahydrofuran (1 mL), and the reaction solution was slowly added dropwise, and after 10 minutes, isoamyl nitrite (315.24 mg, 2.69 mmol) was added, the addition was completed, and the reaction was kept at 5 ° C for 1 hour. The reaction was detected by chromatography with mass spectrometry.
  • reaction solution was quenched with saturated aqueous ammonium chloride solution, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, then filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product of the title compound 455.00mg.
  • the structural representation data is as follows:
  • Step 7 Synthesis of N-(7-amino-4-chloro-3-fluoro-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide
  • the structural representation data is as follows:
  • Step 8 (9H-Fluoren-9-yl)methyl(8-acetamide-5-chloro-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)amino Synthesis of Formate
  • the reaction solution was poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product.
  • the structural representation data is as follows:
  • Step 9 (9H-Fluoren-9-yl)methyl(8-amino-5-chloro-6-fluoro-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl)aminomethyl Synthesis of acid esters
  • the structural representation data is as follows:
  • the structural representation data is as follows:
  • Step eleven (1S,9S)-1-amino-4-chloro-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H- Benzo[de]pyrano[3',4':6,7]indolazino[1,2-b]quinoline-10,13-dione and (1R,9S)-1-amino- 4-Chloro-9-ethyl-5-fluoro-9-hydroxy-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4' Synthesis of :6,7]indolizino[1,2-b]quinoline-10,13-dione
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • Mobile phase A 0.05% acetonitrile
  • mobile phase B water (0.05% formic acid)
  • Step 1 (S)-10-benzyl-23-(2-(methanesulfonyl)pyrimidin-5-yl)-6,9,12,15,18-pentaoxo-3-oxo-5 Synthesis of ,8,11,14,17-pentazaoctadecane-22-ynecarboxylic acid
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • Step 2 N-((S)-10-benzyl-1-((1S,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2 ,3,9,10,13,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolazino[1,2-b]quinoline -1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)-6-( 2-(Methylsulfonyl)pyrimidin-5-yl)hexyl-5-amide and N-((S)-10-benzyl-1-((1R,9S)-4-chloro-9-ethyl- 5-Fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexa
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • Step 3 N-((1S,9S)-4-chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexa Hydro-1H,12H-benzo[de]pyrano[3',4:6,7]indolino[1,2-b]quinolin-1-yl)-2-hydroxyacetamide and N -((1R,9S)-4-Chloro-9-ethyl-5-fluoro-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H, Synthesis of 12H-benzo[de]pyrano[3',4:6,7]indolizino[1,2-b]quinolin-1-yl)-2-hydroxyacetamide
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • Step 1 Synthesis of 1-(4-Fluoro-3-methylphenyl)-3-(isopropylamino)propan-1-one
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • Step 2 Synthesis of 1-(4-Fluoro-5-methyl-2-nitrophenyl)-3-(isopropylamino)propan-1-one
  • the structural representation data is as follows:
  • Step 3 Synthesis of 1-(2-amino-4-fluoro-5-methylphenyl)-3-(isopropylamino)propan-1-one
  • compound 4-12-03 (200.00 mg, 0.75 mmol) was dissolved in methanol (20.0 mL), 10% palladium on carbon (10.00 mg) was added, the reaction solution was replaced with hydrogen, and the reaction was kept at 20 °C in a hydrogen atmosphere. After 16 hours, the reaction was detected by high performance liquid chromatography coupled with mass spectrometry. The reaction solution was filtered and concentrated under reduced pressure to obtain 183.00 mg of the title compound.
  • the structural representation data is as follows:
  • Step 4 (S)-4-ethyl-8-fluoro-4-hydroxy-11-(2-(isopropylamino)ethyl)-9-methyl-1,12-dihydro-14H-pyran Synthesis of Ipo[3',4':6,7]indolazino[1,2-b]quinoline-3,14(4H)-dione
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% trifluoroacetic acid)
  • the structural representation data is as follows:
  • Step 5 (S)-2-((tert-butyldiphenylsilyl)oxy)-N-(2-(4-ethyl-8-fluoro-4-hydroxy-9-methyl-3) ,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-11- Synthesis of yl)ethyl)-N-isopropylacetamide
  • the structural representation data is as follows:
  • Step six (S)-N-(2-(4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro- Synthesis of 1H-pyrano[3',4':6,7]indolazino[1,2-b]quinolin-11-yl)ethyl)-2-hydroxy-N-isopropylacetamide
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • the structural representation data is as follows:
  • the structural representation data is as follows:
  • the reaction solution was slowly poured into saturated aqueous sodium bicarbonate solution, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure to obtain the crude product,
  • the structural representation data is as follows:
  • Step 4 Synthesis of N-(7-bromo-3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide
  • the structural representation data is as follows:
  • Step 5 Synthesis of N-(7-azido-3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide
  • the structural representation data is as follows:
  • Step 6 Synthesis of N-(7-amino-3-fluoro-4,7-dimethyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide
  • the structural representation data is as follows:
  • Step 7 (9H-Fluoren-9-yl)methyl(8-acetamido-6-fluoro-2,5-dimethyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2 -Synthesis of carbamate
  • the structural representation data is as follows:
  • Step 8 (9H-Fluoren-9-yl)methyl(8-amino-6-fluoro-2,5-dimethyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl) ) synthesis of carbamate
  • the structural representation data is as follows:
  • Step 9 (9H-Fluoren-9-yl)methyl((9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2, 3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolazino[1,2-b]quinoline- Synthesis of 1-yl)carbamate
  • the reaction solution was directly The crude product was obtained by evaporation under reduced pressure at 140°C.
  • the structural representation data is as follows:
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • Mobile phase A 0.05% acetonitrile
  • mobile phase B water (0.05% formic acid)
  • Step 1 N-(7-((dimethylamino)methylene)-3-fluoro-4-methyl-8-oxy-5,6,7,8-tetrahydronaphthalene-1-yl)ethyl amide synthesis
  • the structural representation data is as follows:
  • Step 2 Synthesis of N-(7-(aminomethylene)-3-fluoro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalene-1-yl)acetamide
  • the structural representation data is as follows:
  • Step 3 Synthesis of N-(7-(aminomethyl)-3-fluoro-4-methyl-8-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)acetamide
  • the structural representation data is as follows:
  • Step 4 (9H-Fluoren-9-yl)methyl((8-acetamide-6-fluoro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl) ) methyl) carbamate synthesis
  • the reaction solution was poured into water, then extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure to obtain a crude product.
  • the structural representation data is as follows:
  • Step 5 (9H-Fluoren-9-yl)methyl ((8-amino-6-fluoro-5-methyl-1-oxo-1,2,3,4-tetrahydronaphthalene-2-yl) Synthesis of methyl)carbamate
  • the structural representation data is as follows:
  • Step 6 (9H-Fluoren-9-yl)methyl(((9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3, 9,10,13,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolazino[1,2-b]quinoline-1- Synthesis of yl)methyl)carbamate
  • the structural representation data is as follows:
  • Step 7 (1S,9S)-1-(aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro- 10H,13H-benzo[de]pyrano[3',4':6,7]indolazino[1,2-b]quinoline-10,13-dione and (1R,9S)- 1-(Aminomethyl)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de] Synthesis of Pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • Mobile phase A 0.05% acetonitrile
  • mobile phase B water (0.05% formic acid)
  • Step 1 1-((tert-butyldiphenylsilyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl -10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizine [1,2-b]quinolin-1-yl)cyclopropane-1-carboxamide or 1-((tert-butyldiphenylsilyl)oxy)-N-((1R,9S)- 9-Ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo Synthesis of [de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)
  • the structural representation data is as follows:
  • Step 2 N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3,9,10,13 ,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolino[1,2-b]quinolin-1-yl)-1- Hydroxycyclopropane-1-carboxamide or N-((1R,9S)-9-ethyl-5-fluoro-9-hydroxy-1,4-dimethyl-10,13-dioxo-2,3 ,9,10,13,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-1 -Synthesis of -1-hydroxycyclopropane-1-carboxamide
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% formic acid)
  • the structural representation data is as follows:
  • Step 1 (9S)-1-amino-5-chloro-9-ethyl-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo Isolation and Purification of Pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione
  • Mobile phase A acetonitrile
  • mobile phase B water (0.05% trifluoroacetic acid)

Abstract

具有抗肿瘤活性的喜树碱类化合物，及其制备方法和应用。具体地，涉及如下所示化合物或其药学上可接受的形式，其药物组合物、制备方法和用途。所述化合物可用作治疗细胞异常增殖方面疾病的药物，

Description

喜树碱类化合物及其制备方法和应用
本申请是以CN申请号为202110159956.6，申请日为2021年2月5日的申请，CN申请号为202110533304.4，申请日为2021年5月17日的申请，CN申请号为202110718245.8，申请日为2021年6月28日的申请，CN申请号为202110936768.X，申请日为2021年8月16日的申请，以及CN申请号为202111355330.9，申请日为2021年11月16日的申请为基础，并主张其优先权，上述CN申请的公开内容在此作为整体引入本申请中。
技术领域
本发明涉及一类具有抗肿瘤活性的喜树碱类化合物及其缀合物，以及它们的制备方法和在医药领域中的应用。
背景技术
喜树碱(Camptothecin，CPT，式1)是从珙桐科植物喜树(Camptothecaacuminata)中分离得到的一种五环喹啉母核化合物，由喹啉环AB、吡咯环C、吡啶酮环D和α-羟基内酯环E组成，其中20-位为S构型(见下文结构式)。20世纪70年代初由于其优异的抗癌活性而被引入临床，后来由于出现腹泻、出血性膀胱炎等严重副反应，而终止了临床试验。
Figure PCTCN2022074328-appb-000001
研究数据表明，喜树碱可与细胞DNA拓扑异构酶Ⅰ形成三元复合物，从而抑制DNA的解旋，导致DNA复制受阻，进而造成细胞死亡(Cancer Res.1989,49,6365)。喜树碱及其衍生物在肺癌、乳腺癌、结直肠癌、卵巢癌等动物体内模型中具有很强的抗肿瘤活性(Nature Review Cancer.2006,6,789)。
目前已有多个喜树碱类药物被批准上市用于肿瘤治疗(Med.Res.Rev.2015,35,753)。伊立替康为结直肠癌治疗药物；拓扑替康用于卵巢癌的治疗；贝洛替康用于卵巢癌和小细胞肺癌的治疗。喜树碱衍生物还有Exatecan,Rubitecan,Karenitecan,Diflomotecan,Lurtotecan,Gimatecan,Namitecan,Simmitecan,Silatecan,Chimmitecan,Elomotecan等。
喜树碱类药物或衍生物多存在骨髓抑制导致的血液毒性，比如中性粒细胞减少、白细胞减少、血小板减少、贫血等，以及胃肠道副作用，比如恶心、呕吐、腹泻等。临床 研究发现提升喜树碱类化合物安全性和有效性的措施包括改善其药代性质、调节活性、减少用量或者利用其缀合物与抗体形成抗体偶联药物等。因此，研发结构新颖并能够提升有效性、改善安全性问题的喜树碱类化合物及其缀合物仍有很高的临床需求和应用价值。
发明内容
本发明提供结构新颖的喜树碱类化合物及其缀合物，该喜树碱类化合物具有良好的抗肿瘤活性，有望用于肿瘤疾病的治疗；其缀合物有广泛的抗体偶联药物应用前景。
本发明第一方面提供化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，其中所述化合物具有如下所示结构：
Figure PCTCN2022074328-appb-000002
其中，
R 1和R 2各自独立地选自氢、卤素、C 1-6烷基、C 1-6烷氧基、C 1-6卤代烷基、羟基、氰基和C 3-6环烷基；或者，R 1和R 2与相邻碳原子连接成5-6元含氧杂环；
R 3为氢或者与R 1邻位碳原子相连成六元碳环；
A选自
Figure PCTCN2022074328-appb-000003
和
Figure PCTCN2022074328-appb-000004
中的一种；
R 4选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3- 6环烷基和3-6元杂环基；
R 5和R 6各自独立地选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷胺基烷基、C 1-6烷氧基烷基、C 2-6烯基、C 2-6炔基、C 3-6环烷基、3-6元杂环基、3-6元杂环基烷基、芳基和杂芳基；或者R 5和R 6与相邻碳原子连接成3-6元碳环或杂环；
m＝1或2。
在一些实施方案中，所述化合物具有式(I)的结构：
Figure PCTCN2022074328-appb-000005
在式(I)中，R x选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3-6环烷基和3-6元杂环基；
R y和R z不同时为氢，且独立选自氢、C 1-6烷基、C  1-6卤代烷基、C 2-6烯基、C 2-6炔基、C 1-6烷胺基烷基、C 1-6烷氧基烷基、3-6元杂环基烷基和3-6元杂环基。
在一些实施方案中，R x选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3-6环烷基和3-6元杂环基；
R y和R z不同时为氢，且独立选自氢、C 2-6烯基、C 2-6炔基、C 1-6烷胺基烷基、C 1-6烷氧基烷基、3-6元杂环基烷基和3-6元杂环基。
在一些实施方案中，式(I)中，R x选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3-6环烷基、3～6元杂环基；
R y和R z不同时为氢，且独立选自氢、C 2-6烯基、C 2-6炔基。
在一些实施方案中，R x选自氢或C 1-6烷基。
在一些实施方案中，R x为氢。
在一些实施方案中，R y和R z不同时为氢，且独立选自氢、
Figure PCTCN2022074328-appb-000006
二甲胺基亚甲基、吗啉基亚甲基和甲氧基亚甲基。
在一些实施方案中，R y和R z不同时为氢，且独立选自氢、
Figure PCTCN2022074328-appb-000007
二甲胺基亚甲基和甲氧基亚甲基。
在一些实施方案中，R y为氢，R z选自
Figure PCTCN2022074328-appb-000008
二甲胺基亚甲基、吗啉基亚甲基和甲氧基亚甲基。
在一些实施方案中，R y为氢，R z选自
Figure PCTCN2022074328-appb-000009
二甲胺基亚甲基和甲氧基亚甲基。
在一些实施方案中，R x为氢，R y为氢，R z选自
Figure PCTCN2022074328-appb-000010
二甲胺基亚甲基、吗啉基亚甲基和甲氧基亚甲基。
在一些实施方案中，R x为氢，R y为氢，R z选自
Figure PCTCN2022074328-appb-000011
二甲胺基亚甲基和甲氧基亚甲基。
在一些实施方案中，R x为氢，R y为氢，R z选自
Figure PCTCN2022074328-appb-000012
和二甲胺基亚甲基。
在一些实施方案中，式(I)中
Figure PCTCN2022074328-appb-000013
处为
Figure PCTCN2022074328-appb-000014
构型。
在一些实施方案中，式(I)中
Figure PCTCN2022074328-appb-000015
处为
Figure PCTCN2022074328-appb-000016
构型。
在一些实施方案中，式(I)中
Figure PCTCN2022074328-appb-000017
处为
Figure PCTCN2022074328-appb-000018
构型。
在一些实施方案中，式(I)中
Figure PCTCN2022074328-appb-000019
处为
Figure PCTCN2022074328-appb-000020
构型。
在一些实施方案中，式(I)中
Figure PCTCN2022074328-appb-000021
处为
Figure PCTCN2022074328-appb-000022
构型。
在一些实施方案中，式(I)中
Figure PCTCN2022074328-appb-000023
处为
Figure PCTCN2022074328-appb-000024
构型。
在一些实施方案中，式(I)中
Figure PCTCN2022074328-appb-000025
处为
Figure PCTCN2022074328-appb-000026
构型。
在一些实施方案中，式(I)中
Figure PCTCN2022074328-appb-000027
处为
Figure PCTCN2022074328-appb-000028
构型。
在一些实施方案中，所述化合物具有式(II)的结构：
Figure PCTCN2022074328-appb-000029
在式(II)中，A’选自
Figure PCTCN2022074328-appb-000030
和
Figure PCTCN2022074328-appb-000031
中的一种；
R x’选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3- 6环烷基和3-6元杂环基；
R y’和R z’独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3- 6环烷基、3-6元杂环基、3-6元杂环基烷基、C 2-6烯基、C 2-6炔基、芳基和杂芳基，或者R y’和R z’与相邻碳原子连接成3-6元环。
在一些实施方案中，式(II)结构如下式(II)-1所示：
Figure PCTCN2022074328-appb-000032
在一些实施方案中，R x’选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 3- 6环烷基、3～6元杂环基；
R y’和R z’独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 3- 6环烷基、3～6元杂环基、C 2-6烯基、C 2-6炔基、芳基、杂芳基，或者R y’和R z’与相邻碳原子连接成3-6元环。
在一些实施方案中，所述3-6元环选自3-6元碳环或3-6元杂环。
在一些实施方案中，R x’选自氢和C 1-6烷基。
在一些实施方案中，R x’选自氢和甲基。
在一些实施方案中，R y’和R z’独立选自氢、C 1-6烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3- 6环烷基和C 2-6烯基，或者R y’和R z’与相邻碳原子连接成3-6元环烷基。
在一些实施方案中，R y’选自氢和C 1-6烷基，R z’选自氢、C 1-6烷基和C 3- 6环烷基，或者R y’和R z’与相邻碳原子连接成3-6元环。
在一些实施方案中，R y’选自氢和甲基，R z’选自氢、甲基和环丙基，或者R y’和R z’与相邻碳原子连接成3元碳环。
在一些实施方案中，R x’选自氢和甲基，R y’选自氢和甲基，R z’选自氢、甲基和环丙基，或者R y’和R z’与相邻碳原子连接成3元碳环。
在一些实施方案中，R x’为氢，R y’选自氢和甲基，R z’选自氢、甲基和环丙基，或者R y’和R z’与相邻碳原子连接成3元碳环。
在一些实施方案中，式(II)中A’为
Figure PCTCN2022074328-appb-000033
在一些实施方案中，式(II)-1中
Figure PCTCN2022074328-appb-000034
处为
Figure PCTCN2022074328-appb-000035
构型。
在一些实施方案中，式(II)-1中
Figure PCTCN2022074328-appb-000036
处为
Figure PCTCN2022074328-appb-000037
构型。
在一些实施方案中，式(II)-1中
Figure PCTCN2022074328-appb-000038
处为
Figure PCTCN2022074328-appb-000039
构型。
在一些实施方案中，式(II)-1中
Figure PCTCN2022074328-appb-000040
处为
Figure PCTCN2022074328-appb-000041
构型。
在一些实施方案中，式(II)-1中
Figure PCTCN2022074328-appb-000042
处为
Figure PCTCN2022074328-appb-000043
构型。
在一些实施方案中，式(II)-1中
Figure PCTCN2022074328-appb-000044
处为
Figure PCTCN2022074328-appb-000045
构型。
在一些实施方案中，式(II)-1中
Figure PCTCN2022074328-appb-000046
处为
Figure PCTCN2022074328-appb-000047
构型。
在一些实施方案中，式(II)-1中
Figure PCTCN2022074328-appb-000048
处为
Figure PCTCN2022074328-appb-000049
构型。
在一些实施方案中，所述化合物具有式(III)的结构：
Figure PCTCN2022074328-appb-000050
在式(III)中，A”选自
Figure PCTCN2022074328-appb-000051
和
Figure PCTCN2022074328-appb-000052
中的一种；
R x”选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3- 6环烷基和3-6元杂环基；
R y”和R z”独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基、3-6元杂环基、3-6元杂环基烷基、4-6元杂环基、C 2-6烯基、C 2-6炔 基、芳基和杂芳基，或者R y”和R z”与相邻碳原子连接成3-6元环。
在一些实施方案中，式(III)化合物结构如下式(III)-1所示：
Figure PCTCN2022074328-appb-000053
在一些实施方案中，R x”选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 3- 6环烷基、3～6元杂环基；
R y”和R z”独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 3- 6环烷基、4～6元杂环基、C 2-6烯基、C 2-6炔基、芳基、杂芳基，或者R y”和R z”与相邻碳原子连接成3-6元环。
在一些实施方案中，R x”选自氢和C 1-6烷基。
在一些实施方案中，R x”为氢。
在一些实施方案中，所述3-6元环选自3-6元碳环或3-6元杂环。
在一些实施方案中，R y”和R z”各自独立地选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3- 6环烷基和乙烯基，或者R y”和R z”与相邻碳原子连接成3-6元环。
在一些实施方案中，R y”为氢，R z”选自氢、C 1-6烷基、C 3- 6环烷基和乙烯基，或者R y”和R z”与相邻碳原子连接成3-6元碳环。
在一些实施方案中，R y”为氢，R z”选自氢、甲基、环丙基和乙烯基，或者R y”和R z”与相邻碳原子连接成3元碳环。
在一些实施方案中，R x”为氢，R y”为氢，R z”选自氢、甲基、环丙基和乙烯基，或者R y”和R z”与相邻碳原子连接成3元碳环。
在一些实施方案中，式(III)中A”为
Figure PCTCN2022074328-appb-000054
在一些实施方案中，式(III)-1中
Figure PCTCN2022074328-appb-000055
处为
Figure PCTCN2022074328-appb-000056
构型。
在一些实施方案中，式(III)-1中
Figure PCTCN2022074328-appb-000057
处为
Figure PCTCN2022074328-appb-000058
构型。
在一些实施方案中，式(III)-1中
Figure PCTCN2022074328-appb-000059
处为
Figure PCTCN2022074328-appb-000060
构型。
在一些实施方案中，式(III)-1中
Figure PCTCN2022074328-appb-000061
处为
Figure PCTCN2022074328-appb-000062
构型。
在一些实施方案中，式(III)-1中
Figure PCTCN2022074328-appb-000063
处为
Figure PCTCN2022074328-appb-000064
构型。
在一些实施方案中，式(III)-1中
Figure PCTCN2022074328-appb-000065
处为
Figure PCTCN2022074328-appb-000066
构型。
在一些实施方案中，式(III)-1中
Figure PCTCN2022074328-appb-000067
处为
Figure PCTCN2022074328-appb-000068
构型。
在一些实施方案中，式(III)-1中
Figure PCTCN2022074328-appb-000069
处为
Figure PCTCN2022074328-appb-000070
构型。
在一些实施方案中，所述化合物具有式(IV)的结构：
Figure PCTCN2022074328-appb-000071
在式(IV)中，
R a和R b独立选自氢、卤素、C 1-6烷基、C 1-6羟烷基、C 1-6烷氧烷基、C 1-6烷氧基、C 1-6卤代烷基、羟基和氰基；或者R a和R b与相邻碳原子连接成5-6元含氧杂环；
R c和R d独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基、3-6元杂环基、3-6元杂环基烷基、C 2-6烯基和C 2-6炔基，或者R c和R d与相邻碳原子连接成3-6元的碳环或杂环；
R e选自氢、C 1-6烷基、C 3-6环烷基、C 1-6卤代烷基、C 1-6烷氧基烷基和C 2-C 5杂环基；
q＝0或1；
当q＝0时，R c和R d不同时为氢。
在一些实施方案中，R a和R b独立选自氢、卤素、C 1-6烷基、C 1-6烷氧基、C 1-6卤代烷基、羟基和氰基；或者R a和R b与相邻碳原子连接成5-6元含氧杂环；
R c和R d独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基、3-6元杂环基、3-6元杂环基烷基、C 2-6烯基和C 2-6炔基，或者R c和R d与相邻碳原子连接成3-6元的碳环或杂环；
R e选自氢、C 1-6烷基、C 3-6环烷基、C 1-6卤代烷基、C 1-6烷氧基烷基和4-6元杂环基；
q＝0或1；
当q＝0时，R c和R d不同时为氢。
在一些实施方案中，式(IV)中，R a和R b独立选自氢、卤素、C 1-6烷基、C 1-6烷氧基、C 1-6卤代烷基、羟基、和氰基；或者R a和R b与相邻碳原子连接成5-6元含氧杂环；
R c和R d独立选自氢、C 1-6烷基、C 3- 6环烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、4-6元杂环基、C 2-6烯基、C 2-6炔基，或者R c和R d与相邻碳原子连接成3-6元的碳环或杂环；
R e选自氢、C 1-6烷基、C 3- 6环烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、或4-6元杂环基；
q＝0或1；
当q＝0时，R c和R d不同时为氢。
在一些实施方案中，R a和R b独立选自氢、卤素和C 1-6烷基，或者R a和R b与相邻碳原子连接成5-6元含氧杂环。
在一些实施方案中，R a和R b独立选自氢、氟、氯和甲基，或者R a和R b和与其相连的苯环共同形成
Figure PCTCN2022074328-appb-000072
其中Z选自-CH 2-、-CD 2-、-CH 2CH 2-和-CF 2-。
在一些实施方案中，R a为甲基，R b为氟，或者R a和R b和与其相连的苯环共同形成
Figure PCTCN2022074328-appb-000073
在一些实施方案中，R c和R d独立选自氢、
Figure PCTCN2022074328-appb-000074
C 1-6烷氧基烷基和C 1-6烷胺基烷基，或者R c和R d与相邻碳原子连接成3-6元碳环。
在一些实施方案中，R c为氢，R d选自氢、
Figure PCTCN2022074328-appb-000075
甲氧基乙基和环丙基，或者R c和R d与相邻碳原子连接成3-6元碳环。
在一些实施方案中，R e选自氢和C 1-6烷基。
在一些实施方案中，R e选自氢和异丙基。
在一些实施方案中，R a为甲基，R b为氟，或者R a和R b和与其相连的苯环共同形成
Figure PCTCN2022074328-appb-000076
R e选自氢和异丙基，R c为氢，R d选自氢、
Figure PCTCN2022074328-appb-000077
甲氧基乙基和环丙基，或者R c和R d与相邻碳原子连接成3元碳环。
在一些实施方案中，R a为甲基，R b为氟，或者R a和R b和与其相连的苯环共同形成
Figure PCTCN2022074328-appb-000078
R e选自氢和异丙基，R c为氢，R d选自氢、甲氧基乙基和环丙基，或者R c和R d与相邻碳原子连接成3元碳环。
在一些实施方案中，式(IV)中
Figure PCTCN2022074328-appb-000079
处为
Figure PCTCN2022074328-appb-000080
构型。
在一些实施方案中，式(IV)中
Figure PCTCN2022074328-appb-000081
处为
Figure PCTCN2022074328-appb-000082
构型。
在一些实施方案中，所述化合物具有式(V)的结构：
Figure PCTCN2022074328-appb-000083
在式(V)中，R选自C 3-6环烷基和C 1-6烷氧基；
A”’选自
Figure PCTCN2022074328-appb-000084
和
Figure PCTCN2022074328-appb-000085
中的一种；
R x” ’选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3- 6环烷基和3-6元杂环基；
R y” ’和R z” ’独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基、3-6元杂环基、3-6元杂环基烷基、C 2-6烯基、C 2-6炔基、芳基和杂芳基，或者R y” ’和R z” ’与相邻碳原子连接成3-6元环。
在一些实施方案中，R选自甲氧基和环丙基。
在一些实施方案中，式(V)化合物结构如下式(V)-1所示：
Figure PCTCN2022074328-appb-000086
在一些实施方案中，所述3-6元环选自3-6元碳环或3-6元杂环。
在一些实施方案中，R y” ’和R z” ’各自独立地选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基和乙烯基，或者R y” ’和R z” ’与相邻碳原子连接成3-6元碳环。
在一些实施方案中，R y” ’和R z” ’均为氢，或者R y” ’和R z” ’与相邻碳原子连接成3-6元碳环。
在一些实施方案中，R x” ’选自氢和C 1-6烷基。
在一些实施方案中，R x” ’为氢。
在一些实施方案中，A”’为
Figure PCTCN2022074328-appb-000087
在一些实施方案中，R选自甲氧基和环丙基，R x” ’为氢，R y” ’和R z” ’均为氢，或者R y” ’和R z” ’与相邻碳原子连接成3元碳环。
在一些实施方案中，式(V)-1中
Figure PCTCN2022074328-appb-000088
处为
Figure PCTCN2022074328-appb-000089
构型。
在一些实施方案中，式(V)-1中
Figure PCTCN2022074328-appb-000090
处为
Figure PCTCN2022074328-appb-000091
构型。
在一些实施方案中，式(V)-1中
Figure PCTCN2022074328-appb-000092
处为
Figure PCTCN2022074328-appb-000093
构型。
在一些实施方案中，式(V)-1中
Figure PCTCN2022074328-appb-000094
处为
Figure PCTCN2022074328-appb-000095
构型。
在一些实施方案中，式(V)-1中
Figure PCTCN2022074328-appb-000096
处为
Figure PCTCN2022074328-appb-000097
构型。 在一些实施方案中，式(V)-1中
Figure PCTCN2022074328-appb-000098
处为
Figure PCTCN2022074328-appb-000099
构型。在一些实施方案中，式(V)-1中
Figure PCTCN2022074328-appb-000100
处为
Figure PCTCN2022074328-appb-000101
构型。在一些实施方案中，式(V)-1中
Figure PCTCN2022074328-appb-000102
处为
Figure PCTCN2022074328-appb-000103
构型。在一些实施方案中，本发明提供下述化合物：
Figure PCTCN2022074328-appb-000104
Figure PCTCN2022074328-appb-000105
Figure PCTCN2022074328-appb-000106
Figure PCTCN2022074328-appb-000107
Figure PCTCN2022074328-appb-000108
Figure PCTCN2022074328-appb-000109
另一方面，本发明还提供式(VI)化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药：
M-L-E-D
式(VI)
其中，
M是和抗体或其抗原结合片段的接头部位；
L是连接接头M和E之间的连接子；
E是连接L和D的结构片段；
D是细胞毒性药物的结构片段。
在一些实施方案中，M选自以下结构：
Figure PCTCN2022074328-appb-000110
在一些实施方案中，M选自以下结构：
Figure PCTCN2022074328-appb-000111
在一些实施方案中，L选自由下述的一个或多个组成的二价结构：C 1-6亚烷基、-N(R’)-、羰基、-O-、Val、Cit、Phe、Lys、D-Val、Leu、Gly、Ala、Asn、Val-Cit、Val-Ala、Val-Lys、Val-Lys(Ac)、Phe-Lys、Phe-Lys(Ac)、D-Val-Leu-Lys、Gly-Gly-Arg、Ala-Ala-Asn、Ala-Ala-Ala、Val-Lys-Ala、Gly-Gly-Gly、Gly-Gly-Phe-Gly、 Gly-Gly-Gly-Gly-Gly、
Figure PCTCN2022074328-appb-000112
Figure PCTCN2022074328-appb-000113
其中R’代表氢、C 1-6烷基或含-(CH 2CH 2O) r-的烷基；r选自1-10的整数；s选自1-10的整数。
在一些实施方案中，L选自以下结构：
Figure PCTCN2022074328-appb-000114
在一些实施方案中，L选自以下结构：
Figure PCTCN2022074328-appb-000115
在一些实施方案中，E选自单键、-NH-CH 2-、
Figure PCTCN2022074328-appb-000116
在一些实施方案中，E为-NH-CH 2-。
在一些实施方案中，所述细胞毒性药物选自本发明第一方面任一项所述的-化合物。
在一些实施方案中，所述细胞毒性药物选自本发明所述的化合物1-1至1-15；2-1至2-27；3-1至3-26；4-1至4-15；或5-1至5-36。
在一些实施方案中，D选自本发明所述-化合物去氢之后的结构。
在一些实施方案中，D选自本发明所述的化合物1-1至1-15；2-1至2-27；3-1至3-26；4-1至4-15；或5-1至5-36去氢之后的结构。
在一些实施方案中，D选自以下结构：
Figure PCTCN2022074328-appb-000117
在一些实施方案中，D选自以下结构：
Figure PCTCN2022074328-appb-000118
在一些实施方案中，M-L-E-D选自以下化合物：
Figure PCTCN2022074328-appb-000119
Figure PCTCN2022074328-appb-000120
Figure PCTCN2022074328-appb-000121
Figure PCTCN2022074328-appb-000122
Figure PCTCN2022074328-appb-000123
在一些实施方案中，M-L-E-D选自以下化合物：
Figure PCTCN2022074328-appb-000124
Figure PCTCN2022074328-appb-000125
定义
除非在下文中另有定义，本文中所用的所有技术术语和科学术语的含义意图与本领域技术人员通常所理解的相同。提及本文中使用的技术意图指在本领域中通常所理解的技术，包括那些对本领域技术人员显而易见的技术的变化或等效技术的替换。虽然相信以下术语对于本领域技术人员很好理解，但仍然阐述以下定义以更好地解释本发明。
术语“包括”、“包含”、“具有”、“含有”或“涉及”及其在本文中的其它变体形式为包含性的(inclusive)或开放式的，且不排除其它未列举的元素或方法步骤。
如本文中所使用，化合物结构式中标记的“*”表示被标记的碳原子为手性碳原子，本发明包括由该手性碳原子形成的一对对映异构体。如在一个化合物中含两个不同的手性碳原子，本发明包括由该手性碳原子形成的4个旋光异构体。
如本文中所使用，
Figure PCTCN2022074328-appb-000126
表示可以是立体定向((R)或(S))或非立体定向的键。
术语“烷基”定义为直链或支链饱和脂肪族烃基。在一些实施方案中，烷基具有1至12个，例如1至6个碳原子。例如，如本文中所使用，术语“C 1-6烷基”指1至6个碳原子的线性或支化的基团(例如甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基和正己基)，其任选地被1或多个(诸如1、2或3个)适合的取代基取代。
术语“烯基”是指含有至少一个碳碳双键的直链或支链的烃基，包括例如“C 2-6烯基”、“C 2-4烯基”等。其实例包括但不限于：乙烯基、1-丙烯基、2-丙烯基、1-丁烯基、2-丁烯基、1,3-丁二烯基、1-戊烯基、2-戊烯基、3-戊烯基、1,3-戊二烯基、1,4-戊二烯基、1-己烯基、2-己烯基、3-己烯基、1,4-己二烯基等。
术语“炔基”是指含有至少一个碳碳三键的直链或支链烃基。包括例如“C 2-6炔基”、“C 4-6炔基”等。其实例包括但不限于：乙炔基、1-丙炔基、2-丙炔基、1-丁炔基、2-丁炔基、1,3-丁二炔基、1-戊炔基、2-戊炔基、3-戊炔基、1,3-戊二炔基、1,4-戊二炔基、1-己炔基、2-己炔基、3-己炔基、1,4-己二炔基等。
术语“环烷基”是指饱和环状烃基，包括但不限于单环烷基和双环烷基(诸如螺环烷基、并环烷基和桥环烷基)。术语“C 3-6环烷基”指具有3至6个成环碳原子的环烷基，诸如环丙基、环丁基、环戊基、环己基等，其可任选地被1或多个(诸如1、2或3个)适合的取代基取代，例如甲基取代的环丙基。
术语“碳环”或“碳环基”是指饱和或部分不饱和的非芳族单环或多环结构、通过环碳 连接的烃基。其实例包括但不限于环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、环庚基和环辛基。
术语“碳环”指饱和或不饱和的非芳族单环或多环(诸如双环)烃环(例如单环，诸如环丙烷环、环丁烷环、环戊烷环、环己烷环、环庚烷环、环辛烷环、环壬烷环，或双环，包括螺环、稠合或桥连系统(诸如双环[1.1.1]戊烷环、双环[2.2.1]庚烷环、双环[3.2.1]辛烷环或双环[5.2.0]壬烷环、十氢化萘环等)，其可任选地被1或多个(诸如1、2或3个)适合的取代基取代。术语“3-6元碳环”是指含3、4、5或6个成环碳原子的碳环。
术语“杂环基”或“杂环”是指饱和或部分饱和的，单环或多环(诸如双环)的非芳香族环状结构，其环原子由碳原子以及至少一个(例如1、2或3个)选自氮、氧和硫的杂原子构成。如果满足价键要求，杂环基可以通过任意一个环原子与分子的其余部分连接。本发明中的杂环基优选为3-6元杂环基。本发明中所使用的术语“3-6元杂环基”是指具有3至6个环原子的杂环基，包括3元杂环基、4元杂环基、5元杂环基和6元杂环基，包括含氮杂环基、含氧杂环基，例如4-6元杂环基，例如4-6元含氮杂环基、4-6元含氧杂环基。常见的杂环基包括(但不限于)氮杂环丁烷基(azetidinyl)、氧杂环丁烷基(oxetanyl)、四氢呋喃基(tetrahydrofuryl)、吡咯烷基(pyrrolidinyl)、吡咯烷酮基(pyrrolidinonyl)、咪唑烷基(imidazolidinyl)、吡唑烷基(pyrazolidinyl)、四氢吡喃基(tetrahydropyranyl)、哌啶基(piperidinyl)、哌嗪基(piperazinyl)、吗啉基(morpholinyl)。本发明中的杂环基可任选地被一或多个本发明所描述的取代基取代。本发明中的杂环基任选地与一或多个芳香环或非芳香环稠合。
术语“含氧杂环”是指一个或多个(例如1、2或3个)环原子为氧原子的如前所述的杂环，例如5-6元含氧杂环，具体实例包括但不限于环氧乙烷环、四氢呋喃环、呋喃环、四氢吡喃环、吡喃环等。本发明所述的“含氮杂环”是指一个或多个(例如1、2或3个)环原子为氮原子的如前所述的杂环。
术语“卤代烷基”是指被一或多个(诸如1、2或3个)相同或不同的卤素原子取代的烷基，其中烷基定义如上所述。例如，本发明中所使用的术语“C 1-6卤代烷基”是指具有1至6个碳原子的卤代烷基。常见的卤代烷基包括(但不限于)-CH 2F、-CHF 2、-CF 3、-CH 2CF 3、-CF 2CF 3、-CH 2CH 2CF 3、-CH 2Cl等。本发明中的卤代烷基任选地被一或多个本发明所描述的取代基取代。
术语“芳基”是指芳烃分子的芳核碳原子上去掉一个氢原子得到的基团。例如6-14元芳基，具体实例包括但不限于苯基、萘基、蒽基等。
术语“杂芳基”是指含至少一个选自N、O和S的环成员的芳香环状基团。具体实例包括但不限于5-6元杂芳基、5-6元含氮杂芳基、5-6元含氧杂芳基等，例如呋喃基、噻吩基、吡咯基、噻唑基、异噻唑基、噻二唑基、噁唑基、异噁唑基、噁二唑基、咪唑基、吡唑基、1,2,3-三唑基、1,2,4-三唑基、1,2,3-噁二唑基、1,2,4-噁二唑基、1,2,5-噁二唑基、1,3,4-噁二唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基、1,2,3-三嗪基、1,3,5-三嗪 基、1,2,4,5-四嗪基等。
术语“烷氧基”是指具有“烷基-O-”结构的基团，其中烷基定义如上所述。例如C 1-6烷氧基、C 1-4烷氧基、C 1-3烷氧基或C 1-2烷氧基等。常见的烷氧基包括(但不限于)甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、异丁氧基、叔丁氧基、戊氧基、己氧基等。本发明中的烷氧基任选地被一或多个本发明所描述的取代基取代。
()()术语“烷氧基烷基”是指被一个或多个(例如1、2、3或4个)烷氧基取代的烷基，其中烷氧基及烷基的定义如上所述。例如，本发明中所使用的术语“C 1-6烷氧基烷基”是指具有1-6个碳原子、被一个或多个(例如1、2、3或4个)烷氧基取代的烷基。常见的烷氧基烷基包括(但不限于)CH 3O-CH 2-,C 2H 5-O-CH 2-、C 2H 5-O-CH 2CH 2-等。
术语“卤代”或“卤素”基团定义为包括F、Cl、Br或I。
术语“氮氧化物”是指本申请化合物结构中的至少一个氮原子的氧化物(例如单-或二-氧化物)。氮的单-氧化物可以以单一的位置异构体或位置异构体的混合物的形式存在。
术语“取代”指所指定的原子上的一个或多个(例如一个、两个、三个或四个)氢被从所指出的基团的选择代替，条件是未超过所指定的原子在当前情况下的正常原子价并且所述取代形成稳定的化合物。取代基和/或变量的组合仅仅当这种组合形成稳定的化合物时才是允许的。
如果取代基被描述为“任选地被取代”，则取代基可(1)未被取代或(2)被取代。如果取代基的碳被描述为任选地被取代基列表中的一个或多个取代基所取代，则碳上的一个或多个氢(至存在的任何氢的程度)可单独和/或一起被独立地选择的任选的取代基替代。如果取代基的氮被描述为任选地被取代基列表中的一个或多个取代，则氮上的一个或多个氢(至存在的任何氢的程度)可各自被独立地选择的任选的取代基替代。
如果取代基被描述为“独立地选自”一组，则各取代基独立于另一者被选择。因此，各取代基可与另一(其他)取代基相同或不同。
如本文中所使用，术语“一个或多个”意指在合理条件下的1个或超过1个，例如2个、3个、4个、5个或10个。
除非指明，否则如本文中所使用，取代基的连接点可来自取代基的任意适宜位置。
术语“立体异构体”表示由于至少一个不对称中心形成的异构体。在具有一个或多个(例如一个、两个、三个或四个)不对称中心的化合物中，其可产生外消旋混合物、单一对映异构体、非对映异构体混合物和单独的非对映异构体。特定个别分子也可以几何异构体(顺式/反式)存在。类似地，本发明的化合物可以两种或更多种处于快速平衡的结构不同的形式的混合物(通常称作互变异构体)存在。互变异构体的代表性实例包括酮-烯醇互变异构体、苯酚-酮互变异构体、亚硝基-肟互变异构体、亚胺-烯胺互变异构体等。要理解，本申请的范围涵盖所有这样的以任意比例(例如60％、65％、70％、75％、80％、85％、90％、95％、96％、97％、98％或99％)的异构体或其混合物。
本文中可使用实线(—)、实楔形
Figure PCTCN2022074328-appb-000127
或虚楔形
Figure PCTCN2022074328-appb-000128
描绘本发明的化合物的碳-碳键。 使用实线以描绘键连至不对称碳原子的键欲表明，包括该碳原子处的所有可能的立体异构体(例如，特定的对映异构体、外消旋混合物等)。使用实或虚楔形以描绘键连至不对称碳原子的键欲表明，存在所示的立体异构体。当存在于外消旋混合物中时，使用实及虚楔形以定义相对立体化学，而非绝对立体化学。除非另外指明，否则本发明的化合物意欲可以立体异构体(其包括顺式及反式异构体、光学异构体(例如R及S对映异构体)、非对映异构体、几何异构体、旋转异构体、构象异构体、阻转异构体及其混合物)的形式存在。本发明的化合物可表现一种以上类型的异构现象，且由其混合物(例如外消旋混合物及非对映异构体对)组成。
本发明涵盖本发明的化合物的所有可能的结晶形式或多晶型物，其可为单一多晶型物或多于一种多晶型物的任意比例的混合物。
还应当理解，本发明的某些化合物可以游离形式存在用于治疗，或适当时，以其药学上可接受的衍生物形式存在。在本发明中，药学上可接受的衍生物包括但不限于，可药用盐、酯、溶剂合物、代谢物或前药，在将它们向需要其的患者给药后，能够直接或间接提供本发明的化合物或其代谢物或残余物。因此，当在本文中提及“本发明的化合物”时，也意在涵盖化合物的上述各种衍生物形式。
本发明的化合物的可药用盐包括其酸加成盐及碱加成盐。
适合的酸加成盐由形成药学可接受盐的酸来形成，包括天冬氨酸盐、延胡索酸盐、葡庚糖酸盐、葡糖酸盐、葡糖醛酸盐、六氟磷酸盐等。
适合的碱加成盐由形成药学可接受盐的碱来形成，包括铝盐、精氨酸盐、胆碱盐、二乙胺盐等。
适合的盐的综述参见Stahl及Wermuth的“Handbook of Pharmaceutical Salts:Properties,Selection,and Use”(Wiley-VCH,2002)。用于制备本发明的化合物的可药用盐的方法为本领域技术人员已知的。
术语“酯”意指衍生自本申请中各个通式化合物的酯，其包括生理上可水解的酯(可在生理条件下水解以释放游离酸或醇形式的本发明的化合物)。本发明的化合物本身也可以是酯。
本发明的化合物可以溶剂合物(优选水合物)的形式存在，其中本发明的化合物包含作为所述化合物晶格的结构要素的极性溶剂，特别是例如水、甲醇或乙醇。极性溶剂特别是水的量可以化学计量比或非化学计量比存在。
在本发明的范围内还包括本发明的化合物的代谢物，即在给药本发明的化合物时体内形成的物质。这样的产物可由例如被给药的化合物的氧化、还原、水解、酰胺化、脱酰胺化、酯化、脱脂化、酶解等产生。因此，本发明包括本发明的化合物的代谢物，包括通过使本发明的化合物与哺乳动物接触足以产生其代谢产物的时间的方法制得的化合物。
本发明在其范围内进一步包括本发明的化合物的前药。通常这样的前药会是所述化合物的官能团衍生物，其易于在体内转化成期望的治疗活性化合物。因此，在这些情况中， 用于本发明的治疗方法的术语“给药”应包括用所要求保护的化合物中的一种或多种的前药形式来治疗各种疾病或病症，但是在向个体给药后所述前药形式在体内转化成上述化合物。例如，在“Design of Prodrug”,ed.H.Bundgaard,Elsevier,1985中，描述了选择和制备适合的前药衍生物的常规方法。
本发明在其范围内进一步包括本发明的化合物的同位素标记物，其与本发明的化合物相同，除了一个或多个原子被具有相同原子序数但原子质量或质量数不同于在自然界中占优势的原子质量或质量数的原子替代。
本发明还涵盖含有保护基的本发明的化合物。在制备本发明的化合物的任何过程中，保护在任何有关分子上的敏感基团或反应基团可能是必需的和/或期望的，由此形成本发明的化合物的化学保护的形式。这可以通过常规的保护基实现，例如，在Protective Groups in Organic Chemistry,ed.J.F.W.McOmie,Plenum Press,1973；和T.W.Greene&P.G.M.Wuts,Protective Groups in Organic Synthesis,John Wiley&Sons,1991中所述的那些保护基，这些参考文献通过援引加入本文。使用本领域已知的方法，在适当的后续阶段可以移除保护基。
药物组合物
在第三个方面，本发明提供一种药物组合物，其包含本发明第一方面或第二方面所述的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物或前药，以及一种或多种药学上可接受的载体。
术语“药物组合物”是指可以用作药物的组合物，其包含药物活性成分(API)(或治疗剂)以及可选的一种或多种药学上可接受载体。术语“药学上可接受的载体”是指与治疗剂一同给药的辅料，并且其在合理的医学判断的范围内适于接触人类和/或其它动物的组织而没有过度的毒性、刺激性、过敏反应或与合理的益处/风险比相应的其它问题或并发症。
上述药物组合物可以系统地作用和/或局部地作用，其可以通过适合的剂型来实现。所述剂型包括但不限于片剂、胶囊剂、锭剂、硬糖剂、散剂、喷雾剂、乳膏剂、软膏剂、栓剂、凝胶剂、糊剂、洗剂、软膏剂、水性混悬剂、可注射溶液剂、酏剂、糖浆剂。
上述药物组合物可以包含0.01mg至1000mg的至少一种本发明的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物或前药。
本发明还提供了上述药物组合物或其相应的制剂形式的制备方法，其包括将至少一种本发明的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物或前药与一种或多种药学上可接受的载体组合。
药盒产品
在第四个方面，本发明提供一种药盒产品，其包含：
a)作为第一治疗剂的至少一种本发明第一方面或第二方面所述的化合物或其可药用盐、 酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物或前药，或者作为第一药物组合物的第三方面所述的药物组合物；
b)任选存在的作为第二治疗剂的至少一种其他治疗剂，或者作为第二药物组合物的包含其他治疗剂的药物组合物；和
c)任选存在的包装和/或说明书。
上述药盒产品可以包含0.01mg至1000mg的至少一种本发明的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物或前药。
本发明还提供了上述药盒的制备方法，其包括将至少一种本发明的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物或前药或者上述药物组合物与任选存在的至少一种其他治疗剂或者包含其他治疗剂的药物组合物、包装和/或说明书组合。
医药用途
本发明的化合物能够表现出较强的抑制细胞异常增殖方面的作用。
因此，本申请提供了本发明的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药或者上述药物组合物，其用于治疗细胞异常增殖方面的疾病。
另外，本申请还提供了本发明的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药或者上述药物组合物在制备用于治疗细胞异常增殖方面的疾病的药物中的用途。
在一些实施方案中，所述细胞异常增殖方面的疾病包括(但不限于)肿瘤，例如晚期实体瘤。
本申请还提供本发明的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药或者本发明的药物组合物在制备制剂中的用途，所述制剂用于抑制肿瘤细胞的增殖。在某些实施方案中，所述制剂用于体内或者体外施用。例如，所述制剂可被施用至受试者体内，以抑制受试者体内的肿瘤细胞的增殖；或者，所述制剂可被施用至体外细胞(例如细胞系或者来自受试者的细胞)，以抑制体外肿瘤细胞的增殖。
本发明所述肿瘤包括(但不限于)：脑瘤、肺癌、鳞状上皮细胞癌、膀胱癌、胃癌、卵巢癌、腹膜癌、胰腺癌、乳腺癌、头颈癌、子宫颈癌、子宫内膜癌、结直肠癌、肝癌、肾癌、食管腺癌、食管鳞状细胞癌、前列腺癌、雌性生殖道癌、原位癌、淋巴瘤、神经纤维瘤、甲状腺癌、骨癌、皮肤癌、脑癌、结肠癌、睾丸癌、胃肠道间质瘤、前列腺肿瘤、肥大细胞肿瘤、多发性骨髓瘤、黑色素瘤、胶质瘤或肉瘤。
治疗方法
在另一个方面，本发明提供了一种用于治疗细胞异常增殖方面的疾病的方法，其包括下列步骤：将治疗有效量的本发明的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药或者上述药物组合物施用于对其有需求的个体。
术语“有效量”是指能够诱发细胞、组织、器官或生物体(例如个体)产生生物或医学反应，并且足以实现所需预防和/或治疗效果的剂量。
可调整给药方案以提供最佳所需响应。例如，可单次给药，可随时间分剂量给药，或可根据实际情况按比例减少或增加剂量后给药。可以理解的是，对于任何特定个体，具体的给药方案应根据需要以及给药组合物或监督组合物的给药人员的专业判断而调整。
本发明的化合物的给药量将取决于个体情况、疾病或病症的严重性、给药的速率、化合物的处置及处方医师的判断。一般而言，有效量为约0.001-10000mg/kg受试者体重/天。在合适的情况下，有效量为约0.01-1000mg/kg受试者体重/天。可以每天、每两天或每三天给药约0.01-1000mg/kg受试者体重，通常约0.1-500mg/kg受试者体重。示例性的给药方案为每天一次或多次，或者每周一次或多次，或者每月一次或多次。多次给药时，单次剂量之间的间隔通常可以是每天、每周、每月或每年。或者，可以缓释制剂的形式给药，在这种情况下需要较低的给药频率。给药剂量和频率可根据药物在受试者体内的半衰期而不同，也可以根据是预防性应用还是治疗性应用而不同。在预防性应用中，以相对低频率的间隔长期施用相对低的剂量；在治疗性应用中，有时需要以较短的间隔施用相对高的剂量，直至疾病的进展被延缓或停止，优选直至个体表现出疾病症状的部分或完全改善，此后可以采用预防性应用。
术语“治疗”是指减轻或消除所针对的疾病或病症。如果受试者接受了治疗量的本发明的化合物或其药学上可接受的形式或者本发明的药物组合物，该受试者的至少一种指标和症状表现出可观察到的和/或可检测出的缓解和/或改善，则表明该受试者已被成功地“治疗”。可以理解的是，治疗不仅包括完全地治疗，还包括未达到完全地治疗，但实现了一些生物学或医学相关的结果。
术语“施用(administrate/administrating/administration)”(或“给药”)是指将药物活性成分(比如本发明的化合物)或包含药物活性成分的药物组合物(例如本发明的药物组合物)应用于个体或其细胞、组织、器官、生物流体等部位，以便使药物活性成分或药物组合物与个体或其细胞、组织、器官、生物流体等部位接触的过程。常见的施用方式包括(但不限于)口服施用、皮下施用、肌内施用、腹膜下施用、眼部施用、鼻部施用、舌下施用、直肠施用、阴道施用等。
术语“对其有需求”是指医生或其他护理人员对个体需要或者将要从预防和/或治疗过程中获益的判断，该判断的得出基于医生或其他护理人员在其专长领域中的各种因素。
术语“个体”(或称受试者)是指人类或非人动物。本发明的个体包括患有疾病和/或病症的个体(患者)和正常的个体。本发明的非人动物包括所有脊椎动物，例如非哺乳动物，例 如鸟类、两栖类、爬行类等，和哺乳动物，例如非人灵长类、家畜和/或驯化动物(例如绵羊、犬、猫、奶牛、猪等)。
制备方法
本发明第四方面提供了所述化合物的合成方法。
本发明中的式(I)化合物可由下述合成路线进行合成制备。
Figure PCTCN2022074328-appb-000129
其中，R x、R y和R z含义如前文所述；LG为离去基团，选自甲磺酰基、三氟甲磺酰氧基和卤素，优选三氟甲磺酰氧基或碘；
步骤一：
通过式(I)-SM1化合物和式(I)-SM2化合物发生取代反应得到式(I)-IM1化合物。
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选50℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自卤代烃类(例如二氯甲烷(DCM)、氯仿(TCM)、1,2-二氯乙烷(1,2-DCE)等)、腈类(例如乙腈(AN)等)、N-甲基吡咯烷酮(NMP)、N,N-二甲基甲酰胺(DMF)、N,N-二甲基乙酰胺(DMA)、四氢呋喃(THF)、1,4-二氧六环(Dioxane)、二甲基亚砜(DMSO)及其任意组合，优选乙腈。
在部分实施方案中，该步骤在适合的碱存在下进行，所述碱包括有机碱或无机碱，所述有机碱可选自N,N-二异丙基乙胺(DIPEA)、三乙胺(TEA)、叔丁醇钾(t-BuOK)和吡啶(Py)，所述无机碱可选自磷酸钾(K 3PO4)、氢化钠(NaH)、碳酸钾(K 2CO 3)、碳酸钠(Na 2CO 3)、碳酸氢钠(NaHCO 3)、碳酸铯(Cs 2CO 3)和NaOH，优选Na 2CO 3或NaHCO 3；
步骤二：
通过式(I)-IM1化合物和式(I)-SM3化合物发生缩合反应得到式(I)化合物；
在部分实施方案中，该步骤在合适的缩合试剂下进行，所述缩合试剂可选自HATU、HBTU、EDCI、DCC和HOBT，优选HATU；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、 正己烷、乙酸乙酯，优选N,N-二甲基甲酰胺。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选DIPEA。
本发明中的式(II)-1化合物可由下述合成路线进行合成制备。
Figure PCTCN2022074328-appb-000130
其中，R x’、R y’和R z’含义如前文所述；LG为离去基团，选自甲磺酰基、三氟甲磺酰氧基和卤素，优选三氟甲磺酰氧基或碘；PG为保护基，选自
Figure PCTCN2022074328-appb-000131
Figure PCTCN2022074328-appb-000132
步骤一
通过式(II)-SM1化合物发生取代反应得到式(II)-IM1化合物。
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、100℃，优选50℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选正庚烷。
步骤二
通过式(II)-IM1化合物发生还原反应得到式(II)-IM2化合物；
在部分实施方案中，该步骤在合适的还原剂存在下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选铂催化剂；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、100℃，优选60℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选乙酸乙酯。
步骤三
通过式(II)-IM2化合物发生取代反应得到式(II)-IM3化合物，
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、100℃，优选20℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选乙酸乙酯；
在部分实施方案中，该步骤在碱性条件下进行，提供碱性条件的试剂包括有机碱和无机碱类，所述有机碱类包括但不限于三乙胺、吡啶、N,N-二异丙基乙胺、正丁基锂、二异丙基氨基锂、双三甲基硅基胺基锂、双三甲基硅基胺基钠；所述无机碱包括但不限于碳酸钾、碳酸钠、碳酸氢钠、叔丁醇钾、氢化钠、氢氧化钠、氢氧化钾，优选三乙胺。
步骤四
通过式(II)-IM3化合物发生偶联反应得到式(II)-IM4化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、70℃、100℃，优选70℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯、水，优选四氢呋喃和水混合溶剂；
在部分实施方案中，该步骤在碱性条件下进行，提供碱性条件的试剂包括有机碱和无机碱类，所述有机碱类包括但不限于三乙胺、吡啶、N,N-二异丙基乙胺、正丁基锂、二异丙基氨基锂、双三甲基硅基胺基锂、双三甲基硅基胺基钠；所述无机碱包括但不限于碳酸钾、碳酸钠、碳酸氢钠、叔丁醇钾、氢化钠、氢氧化钠、氢氧化钾，优选N,N-二异丙基乙胺。
步骤五
通过式(II)-IM4化合物发生还原反应得到式(II)-IM5化合物；
在部分实施方案中，该步骤在合适的还原剂下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选铂催化剂；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选40℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选四氢呋喃。
步骤六
通过式(II)-IM5化合物发生关环反应得到式(II)-IM6化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为5℃、20℃、25℃、40℃、50℃、60℃、100℃，优选5℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自三氟乙酸、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，叔丁醇，优选四氢呋喃和叔丁醇的混合溶剂。
步骤七
通过式(II)-IM6化合物发生取代反应得到式(II)-IM7化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为5℃、20℃、25℃、40℃、50℃、60℃、100℃，优选5℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自三氟乙酸、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选三氟乙酸；
在部分实施方案中，该步骤在碱性条件下进行，提供碱性条件的试剂包括有机碱和无机碱类，所述有机碱类包括但不限于三乙胺、吡啶、N,N-二异丙基乙胺、正丁基锂、二异丙基氨基锂、双三甲基硅基胺基锂、双三甲基硅基胺基钠；所述无机碱包括但不限于碳酸钾、碳酸钠、碳酸氢钠、叔丁醇钾、氢化钠、氢氧化钠、氢氧化钾，优选叔丁醇钾。
步骤八
通过式(II)-IM7化合物发生还原反应得到式(II)-IM8化合物；
在部分实施方案中，该步骤在合适的还原剂下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选钯催化剂；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选20℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选甲醇。
步骤九
通过式(II)-IM8化合物发生取代反应得到式(II)-IM9化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为5℃、20℃、25℃、40℃、50℃、60℃、100℃，优选20℃；
在部分实施方案中，该步骤在碱性条件下进行，提供碱性条件的试剂包括有机碱和无机碱类，所述有机碱类包括但不限于三乙胺、吡啶、N,N-二异丙基乙胺、正丁基锂、二异丙基氨基锂、双三甲基硅基胺基锂、双三甲基硅基胺基钠；所述无机碱包括但不限于碳酸钾、碳酸钠、碳酸氢钠、叔丁醇钾、氢化钠、氢氧化钠、氢氧化钾，优选吡啶。
步骤十
通过式(II)-IM9化合物发生水解反应得到式(II)-IM10化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选60℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选甲醇。
在部分实施方案中，所述反应在酸性条件下进行，提供酸性条件的试剂包括盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为盐酸。
步骤十一
通过式(II)-IM10化合物和(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃O[3,4-F]吲哚嗪-3,6,10(4H)-酮发生环合反应得到式(II)-IM11化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、 50℃、60℃、100℃、140℃，优选140℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲苯、甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选甲苯。
在部分实施方案中，所述反应在酸性条件下进行，提供酸性条件的试剂包括对甲苯磺酸、盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为对甲苯磺酸。
步骤十二
通过式(II)-IM11化合物发生水解反应得到式(II)-IM12化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选100℃；
在部分实施方案中，所述反应在酸性条件下进行，提供酸性条件的试剂包括对甲苯磺酸、盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为盐酸。
步骤十三
通过式(II)-IM12化合物和式(II)-SM2化合物和发生取代反应得到式(II)-IM13化合物。
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选50℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自卤代烃类(例如二氯甲烷(DCM)、氯仿(TCM)、1,2-二氯乙烷(1,2-DCE)等)、腈类(例如乙腈(AN)等)、N-甲基吡咯烷酮(NMP)、N,N-二甲基甲酰胺(DMF)、N,N-二甲基乙酰胺(DMA)、四氢呋喃(THF)、1,4-二氧六环(Diox)、二甲基亚砜(DMSO)及其任意组合，优选乙腈。
在部分实施方案中，该步骤在适合的碱存在下进行，所述碱包括有机碱或无机碱，所述有机碱可选自N,N-二异丙基乙胺(DIPEA)、三乙胺(TEA)、叔丁醇钾(t-BuOK)和吡啶(Py)，所述无机碱可选自磷酸钾(K 3PO 4)、氢化钠(NaH)、碳酸钾(K 2CO 3)、碳酸钠(Na 2CO 3)、碳酸氢钠(NaHCO 3)、碳酸铯(Cs 2CO 3)和NaOH，优选Na 2CO 3或NaHCO 3；
步骤十四
通过式(II)-IM13化合物和式(II)-SM3化合物发生缩合反应得到式(II)-IM14化合物；
在部分实施方案中，该步骤在合适的缩合试剂下进行，所述缩合试剂可选自HATU、HBTU、EDCI、DCC和HOBT，优选HATU；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选N,N-二甲基甲酰胺。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述 有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选DIPEA。
步骤十五
通过式(II)-IM14化合物的酸解反应得到式(II)-1化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯及其混合溶剂，优选二氯甲烷和甲醇的混合溶液(体积比2：1)。
在部分实施方案中，所述反应在酸性条件下进行，提供酸性条件的试剂包括对甲苯磺酸、盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为盐酸。
式(II)-SM3合成方法如下所示：
当PG为
Figure PCTCN2022074328-appb-000133
时：
Figure PCTCN2022074328-appb-000134
步骤一
通过式(II)-SM3-1化合物和式(II)-SM3-2化合物发生取代反应得到式(II)-SM3-3化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选0-25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯及其混合溶剂，优选四氢呋喃。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选K 2CO 3。
步骤二
通过式(II)-SM3-3化合物发生氢化反应得到式(II)-SM3化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯及其混合溶剂，优选甲醇。
在部分实施方案中，该步骤在合适的还原剂下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选钯催化剂；
当PG为
Figure PCTCN2022074328-appb-000135
时，R y’和R z’为氢：
Figure PCTCN2022074328-appb-000136
步骤一
通过式(II)-SM3-4化合物和式(II)-SM3-5化合物发生缩合反应得到式(II)-SM3化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯及其混合溶剂，优选N,N-二甲基甲酰胺。
或者,式(II)-1化合物可由下述合成路线进行合成制备：
Figure PCTCN2022074328-appb-000137
步骤一
通过式(II)-IM13化合物和式(II)-SM4化合物发生缩合反应得到式(II)-IM15化合物；
在部分实施方案中，该步骤在合适的缩合试剂下进行，所述缩合试剂可选自HATU、HBTU、EDCI、DCC和HOBT，优选HATU；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选N,N-二甲基甲酰胺。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选DIPEA。
步骤二
通过式(II)-IM15化合物脱除硅保护基得到式(II)-1化合物；
或者,式(II)化合物可由下述合成路线进行合成制备：
Figure PCTCN2022074328-appb-000138
步骤一
通过式(II)-IM13化合物和式(II)-SM5化合物发生缩合反应得到式(II)化合物；
在部分实施方案中，该步骤在合适的缩合试剂下进行，所述缩合试剂可选自HATU、 HBTU、EDCI、DCC和HOBT，优选HATU；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选N,N-二甲基甲酰胺。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选DIPEA。
本发明中的式(III)-1化合物可由下述合成路线进行合成制备。
Figure PCTCN2022074328-appb-000139
其中，R x”、R y”和R z”含义如前文所述；LG为离去基团，选自甲磺酰基、三氟甲磺酰氧基和卤素，优选三氟甲磺酰氧基或氯；PG为保护基，选自
Figure PCTCN2022074328-appb-000140
Figure PCTCN2022074328-appb-000141
步骤一
通过式(III)-SM1化合物发生取代反应得到式(III)-IM1化合物。
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、100℃，优选50℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选正庚烷。
步骤二
通过式(III)-IM1化合物发生还原反应得到式(III)-IM2化合物；
在部分实施方案中，该步骤在合适的还原剂存在下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选铂催化剂；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、100℃，优选60℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选乙酸乙酯。
步骤三
通过式(III)-IM2化合物发生取代反应得到式(III)-IM3化合物，
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、100℃，优选20℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选乙酸乙酯；
在部分实施方案中，该步骤在碱性条件下进行，提供碱性条件的试剂包括有机碱和无机碱类，所述有机碱类包括但不限于三乙胺、吡啶、N,N-二异丙基乙胺、正丁基锂、二异丙基氨基锂、双三甲基硅基胺基锂、双三甲基硅基胺基钠；所述无机碱包括但不限于碳酸钾、碳酸钠、碳酸氢钠、叔丁醇钾、氢化钠、氢氧化钠、氢氧化钾，优选三乙胺。
步骤四
通过式(III)-IM3化合物发生偶联反应得到式(III)-IM4化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、70℃、100℃，优选70℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯、水，优选四氢呋喃和水混合溶剂；
在部分实施方案中，该步骤在碱性条件下进行，提供碱性条件的试剂包括有机碱和无机碱类，所述有机碱类包括但不限于三乙胺、吡啶、N,N-二异丙基乙胺、正丁基锂、二异丙基氨基锂、双三甲基硅基胺基锂、双三甲基硅基胺基钠；所述无机碱包括但不限于碳酸钾、碳酸钠、碳酸氢钠、叔丁醇钾、氢化钠、氢氧化钠、氢氧化钾，优选N,N-二异丙基乙胺。
步骤五
通过式(III)-IM4化合物发生还原反应得到式(III)-IM5化合物；
在部分实施方案中，该步骤在合适的还原剂下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选铂催化剂；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选40℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选四氢呋喃。
步骤六
通过式(III)-IM5化合物发生关环反应得到式(III)-IM6化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为5℃、20℃、25℃、40℃、50℃、60℃、100℃，优选5℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自三氟乙酸、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，叔丁醇，优选四氢呋喃和叔丁醇的混合溶剂。
步骤七
通过式(III)-IM6化合物发生取代反应得到式(III)-IM7化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为5℃、20℃、25℃、40℃、50℃、60℃、100℃，优选5℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自三氟乙酸、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选三氟乙酸；
在部分实施方案中，该步骤在碱性条件下进行，提供碱性条件的试剂包括有机碱和无机碱类，所述有机碱类包括但不限于三乙胺、吡啶、N,N-二异丙基乙胺、正丁基锂、二异丙基氨基锂、双三甲基硅基胺基锂、双三甲基硅基胺基钠；所述无机碱包括但不限于碳酸钾、碳酸钠、碳酸氢钠、叔丁醇钾、氢化钠、氢氧化钠、氢氧化钾，优选叔丁醇钾。
步骤八
通过式(III)-IM7化合物发生还原反应得到式(III)-IM8化合物；
在部分实施方案中，该步骤在合适的还原剂下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选钯催化剂；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选20℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选甲醇。
步骤九
通过式(III)-IM8化合物发生取代反应得到式(III)-IM9化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为5℃、20℃、25℃、40℃、50℃、60℃、100℃，优选20℃；
在部分实施方案中，该步骤在碱性条件下进行，提供碱性条件的试剂包括有机碱和无机碱类，所述有机碱类包括但不限于三乙胺、吡啶、N,N-二异丙基乙胺、正丁基锂、二异丙基氨基锂、双三甲基硅基胺基锂、双三甲基硅基胺基钠；所述无机碱包括但不限于碳酸钾、碳酸钠、碳酸氢钠、叔丁醇钾、氢化钠、氢氧化钠、氢氧化钾，优选吡啶。
步骤十
通过式(III)-IM9化合物发生水解反应得到式(III)-IM10化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选60℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选甲醇。
在部分实施方案中，所述反应在酸性条件下进行，提供酸性条件的试剂包括盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为盐酸。
步骤十一
通过式(III)-IM10化合物和(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃O[3,4-F]吲哚嗪-3,6,10(4H)-酮发生环合反应得到式(III)-IM11化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、 50℃、60℃、100℃、140℃，优选140℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲苯、甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选甲苯。
在部分实施方案中，所述反应在酸性条件下进行，提供酸性条件的试剂包括对甲苯磺酸、盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为对甲苯磺酸。
步骤十二
通过式(III)-IM11化合物发生水解反应得到式(III)-IM12化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选100℃；
在部分实施方案中，所述反应在酸性条件下进行，提供酸性条件的试剂包括对甲苯磺酸、盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为盐酸。
步骤十三
通过式(III)-IM12化合物和式(III)-SM2化合物和发生取代反应得到式(III)-IM13化合物。
发生取代反应得到式(II)-IM13化合物。
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选50℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自卤代烃类(例如二氯甲烷(DCM)、氯仿(TCM)、1,2-二氯乙烷(1,2-DCE)等)、腈类(例如乙腈(AN)等)、N-甲基吡咯烷酮(NMP)、N,N-二甲基甲酰胺(DMF)、N,N-二甲基乙酰胺(DMA)、四氢呋喃(THF)、1,4-二氧六环(Dioxane)、二甲基亚砜(DMSO)及其任意组合，优选乙腈。
在部分实施方案中，该步骤在适合的碱存在下进行，所述碱包括有机碱或无机碱，所述有机碱可选自N,N-二异丙基乙胺(DIPEA)、三乙胺(TEA)、叔丁醇钾(t-BuOK)和吡啶(Py)，所述无机碱可选自磷酸钾(K 3PO 4)、氢化钠(NaH)、碳酸钾(K 2CO 3)、碳酸钠(Na 2CO 3)、碳酸氢钠(NaHCO 3)、碳酸铯(Cs 2CO 3)和NaOH，优选Na 2CO 3或NaHCO 3；
步骤十四
通过式(III)-IM13化合物和式(III)-SM3化合物发生缩合反应得到式(III)-IM14化合物；
在部分实施方案中，该步骤在合适的缩合试剂下进行，所述缩合试剂可选自HATU、HBTU、EDCI、DCC和HOBT，优选HATU；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、 四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选N,N-二甲基甲酰胺。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选DIPEA。
步骤十五
通过式(III)-IM14化合物通过酸解反应得到式(III)-1化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯及其混合溶剂，优选二氯甲烷和甲醇的混合溶液(体积比2：1)。
在部分实施方案中，所述反应在酸性条件下进行，提供酸性条件的试剂包括对甲苯磺酸、盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为盐酸。
式(III)-SM3合成方法如下所示：
当PG为
Figure PCTCN2022074328-appb-000142
时：
Figure PCTCN2022074328-appb-000143
步骤一：
通过式(III)-SM3-1化合物和式(II)-SM3-2化合物发生取代反应得到式(III)-SM3-3化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选0-25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯及其混合溶剂，优选四氢呋喃。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选K 2CO 3。
步骤二：
通过式(III)-SM3-3化合物发生氢化反应得到式(III)-SM3化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯及其混合溶剂，优选甲醇。
在部分实施方案中，该步骤在合适的还原剂下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选钯催化剂；
当PG为
Figure PCTCN2022074328-appb-000144
时，R y’和R z’为氢：
Figure PCTCN2022074328-appb-000145
步骤一：
通过式(III)-SM3-4化合物和式(III)-SM3-5化合物发生缩合反应得到式(III)-SM3化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯及其混合溶剂，优选N,N-二甲基甲酰胺。
或者,式(III)-1化合物可由下述合成路线进行合成制备：
Figure PCTCN2022074328-appb-000146
步骤一：
通过式(III)-IM13化合物和式(III)-SM4化合物发生缩合反应得到式(III)-IM15化合物；
在部分实施方案中，该步骤在合适的缩合试剂下进行，所述缩合试剂可选自HATU、HBTU、EDCI、DCC和HOBT，优选HATU；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选N,N-二甲基甲酰胺。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选DIPEA。
步骤二：
通过式(III)-IM15化合物脱除硅保护基得到式(III)-1化合物；
或者,式(III)-1化合物可由下述合成路线进行合成制备：
Figure PCTCN2022074328-appb-000147
步骤一：
通过式(III)-IM13化合物和式(III)-SM5化合物发生缩合反应得到式(III)-1化合物；
在部分实施方案中，该步骤在合适的缩合试剂下进行，所述缩合试剂可选自HATU、HBTU、EDCI、DCC和HOBT，优选HATU；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选N,N-二甲基甲酰胺。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选DIPEA。
本发明中的式(IV)化合物可由下述合成路线进行合成制备：
其中，R a、R b、R c、R d和R e含义如前文所述；
当q＝1时，
Figure PCTCN2022074328-appb-000148
步骤一
通过式(IV)-SM1化合物发生硝化反应得到式(IV)-IM1化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为5℃、20℃、25℃、40℃、50℃、60℃、100℃，优选25℃。
步骤二
通过式(IV)-IM1化合物发生氢化反应得到式(IV)-IM2化合物；
在部分实施方案中，该步骤在合适的还原剂下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选钯催化剂；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选乙酸乙酯。
步骤三
通过式(IV)-IM2化合物发生酰化反应得到式(IV)-IM3化合物，
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、100℃，优选25℃；
步骤四
通过式(IV)-IM3化合物与DMF-DMA反应得到式(IV)-IM4化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、120℃，优选120℃；
步骤五
通过式(IV)-IM4和式(IV)-SM2化合物发生取代反应得到式(IV)-IM5化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选50℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、乙醇、N、N-甲基吡咯烷酮、二甲基亚砜，优选乙醇；
步骤六
通过式(IV)-IM5化合物发生还原反应得到式(IV)-IM6化合物；
在部分实施方案中，该步骤在合适的还原剂下进行，所述还原剂优选硼氢化钠；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选0-25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、冰醋酸、甲醇及其混合溶液，优选冰醋酸。
步骤七
通过式(IV)-IM6化合物的氨基用Fmoc保护得到式(IV)-IM7化合物；
步骤八
通过式(IV)-IM7化合物的氨基脱除乙酰基保护基得到式(IV)-IM8化合物；
步骤九
通过式(IV)-IM8化合物在酸性条件下发生闭环反应得到式(IV)-IM9化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、120℃，优选120℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲苯、 二甲苯、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜，优选甲苯和二甲苯；
在部分实施方案中，该步骤在酸性条件下进行；
提供酸性条件的试剂包括对甲苯磺酸、盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为对甲苯磺酸。
步骤十
通过式(IV)-IM9化合物脱除Fmoc保护基得到式(IV)-IM10化合物；
步骤十一
通过式(IV)-IM10化合物和式(IV)-SM4化合物发生缩合反应得到式(IV)化合物；
在部分实施方案中，该步骤在合适的缩合试剂下进行，所述缩合试剂可选自HATU、HBTU、EDCI、DCC和HOBT，优选HBTU；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选N,N-二甲基甲酰胺。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选DIPEA。
或者，当q＝0时，
Figure PCTCN2022074328-appb-000149
LG为离去基团，选自甲磺酰基、三氟甲磺酰氧基和卤素，优选三氟甲磺酰氧基或碘；
步骤一
通过式(IV)-SM5化合物发生还原反应得到式(IV)-IM11化合物，
在部分实施方案中，该步骤在合适的还原剂存在下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂，优选铂催化剂；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选乙酸乙酯和四氢呋喃。
步骤二
通过式(IV)-IM11化合物发生傅克酰基化反应得到式(IV)-IM12化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为5℃、20℃、25℃、40℃、50℃、60℃、100℃，优选25℃。
步骤三
通过式(IV)-IM12在酸性条件下发生闭环反应得到式(IV)-IM13化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、120℃，优选120℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲苯、二甲苯、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜，优选甲苯和二甲苯；
在部分实施方案中，该步骤在酸性条件下进行；
提供酸性条件的试剂包括对甲苯磺酸、盐酸、三氟乙酸、甲酸、硫酸、甲磺酸，优选为对甲苯磺酸。
步骤四
通过式(IV)-IM13发生取代反应得到式(IV)-IM14化合物；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、乙醇、N、N-甲基吡咯烷酮、二甲基亚砜，优选二甲基亚砜；
步骤五
通过式(IV)-IM14化合物发生还原反应得到式(IV)-IM15化合物，
在部分实施方案中，该步骤在合适的还原剂存在下进行，所述还原剂可选自钯催化剂、铂催化剂、铑催化剂、三苯基膦、亚磷酸三乙酯，优选亚磷酸三乙酯；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、50℃、60℃、80℃、100℃，优选80℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯、甲苯、及其混合溶液，优选甲醇和甲苯的混合溶液。
步骤六
通过式(IV)-IM15化合物和式(IV)-SM6化合物发生取代反应得到式(IV)-IM16化合物。
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃、140℃，优选50℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自卤代烃类(例如二氯甲烷(DCM)、氯仿(TCM)、1,2-二氯乙烷(1,2-DCE)等)、腈类(例如乙腈(AN)等)、N-甲基吡咯烷酮(NMP)、N,N-二甲基甲酰胺(DMF)、N,N-二甲基乙酰胺(DMA)、四氢呋喃(THF)、1,4-二氧六环(Dioxane)、二甲基亚砜(DMSO)及其任意组合，优选乙腈。
在部分实施方案中，该步骤在适合的碱存在下进行，所述碱包括有机碱或无机碱，所述有机碱可选自N,N-二异丙基乙胺(DIPEA)、三乙胺(TEA)、叔丁醇钾(t-BuOK)和 吡啶(Py)，所述无机碱可选自磷酸钾(K 3PO4)、氢化钠(NaH)、碳酸钾(K 2CO 3)、碳酸钠(Na 2CO 3)、碳酸氢钠(NaHCO 3)、碳酸铯(Cs 2CO 3)和NaOH，优选Na 2CO 3或NaHCO 3；
步骤七
通过式(IV)-IM16化合物和式(IV)-SM4化合物发生缩合反应得到式(IV)化合物；
在部分实施方案中，该步骤在合适的缩合试-剂下进行，所述缩合试剂可选自HATU、HBTU、EDCI、DCC和HOBT，优选HBTU；
在部分实施方案中，该步骤在适合的温度下进行，所述温度为20℃、25℃、40℃、50℃、60℃、100℃，优选25℃；
在部分实施方案中，该步骤在适合的有机溶剂中进行，所述有机溶剂可选自甲醇、四氢呋喃、二氯甲烷、N,N-二甲基甲酰胺、N-甲基吡咯烷酮、二甲基亚砜、正庚烷、正己烷、乙酸乙酯，优选N,N-二甲基甲酰胺。
在部分实施方案中，该步骤在适合的碱中进行，所述碱包括有机碱或无机碱，所述有机碱可选自DIPEA、TEA、t-BuOK和Py，所述无机碱可选自K 3PO 4、NaH、K 2CO 3、Na 2CO 3、Cs 2CO 3和NaOH，优选DIPEA。
本发明中的式(V)-1化合物可使用起始物料
Figure PCTCN2022074328-appb-000150
按照式(III)同理的合成路线进行合成制备。
发明的有益效果
本发明提供式(I)-式(IV)所示的喜树碱类化合物，及其药物组合物、制备方法和用途。该类化合物具有良好的抗肿瘤活性，有克服耐药的潜力，可用于治疗细胞异常增殖性病症，所述病症包括但不限于晚期实体瘤。
具体实施方式
以下通过具体实施方式的描述对本申请作进一步说明，但这并非是对本申请的限制。本领域技术人员根据本申请的教导，可以做出各种修改或改进，而不脱离本申请的基本思想和范围。
本发明中的缩写具有以下含义：
Figure PCTCN2022074328-appb-000151
Figure PCTCN2022074328-appb-000152
以下的实施例中记载的化合物的结构通过核磁共振( 1H NMR)或质谱(MS)来确定。
核磁共振( 1H NMR)的测定仪器使用Bruker 400 MHz核磁共振仪；六氘代二甲基亚砜(DMSO-d 6)；内标物质为四甲基硅烷(TMS)。
实施例中使用的核磁共振(NMR)图谱中的缩写示于以下。
s：单峰(singlet)、d：二重峰(doublet)、t：三重峰(triplet)、q：四重峰(quartet)、m：多重峰(multiplet)、br：宽峰(broad)、J：偶合常数、Hz：赫兹、DMSO-d6：氘化二甲基亚砜。δ值用ppm值表示。
质谱(MS)的测定仪器使用Agilent(ESI)质谱仪，型号为Agilent 6120B。
实施例一(R)-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基戊-3-炔酰胺和(S)-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基戊-3-炔酰胺
Figure PCTCN2022074328-appb-000153
将化合物2-羟基戊-3-炔酸(4.29mg，37.63μmol)用DMF(1mL)溶解，加入HATU(21.46mg,56.44μmol)、SM1-1(10.00mg，22.94μmol)和DIPEA(7.29mg,56.44μmol)，25℃反应2小时。反应液经减压浓缩，浓缩物直接经制备高效液相色谱纯化(条件如下)得标题化合物1-1-A 3.24mg和1-1-B 3.98mg。
色谱柱:SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A:乙腈；流动相B:水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 30 70 28
18 90 10 28
保留时间：1-1-A：10.8min；1-1-B：11.1min.
1-1-A结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.58(d,J＝8.4Hz,1H),7.78(d,J＝11.2Hz,1H),7.31(s,1H),6.54(s,1H),6.17(d,J＝6.0Hz,1H),5.58-5.47(m,1H),5.42(s,2H),5.23(s,2H),4.73-4.64(m,1H),3.25-3.06(m,2H),2.39(s,3H),2.27-2.05(m,2H),1.96-1.77(m,5H),0.87(t,J＝7.2Hz,3H).
ESI-MS(m/z):532.2[M+H] +.
1-1-B结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.62(d,J＝8.8Hz,1H),7.78(d,J＝10.8Hz,1H),7.31(s,1H),6.54(s,1H),6.19(d,J＝6.0Hz,1H),5.58-5.47(m,1H),5.42(s,2H),5.21(d,J＝4.8Hz,2H),4.73-4.65(m,1H),3.27-3.06(m,2H),2.39(s,3H),2.27-2.05(m,2H),1.93-1.80(m,2H),1.80(d,J＝2.0Hz,3H),0.87(t,J＝7.2Hz,3H).
ESI-MS(m/z):532.2[M+H] +.
实施例二(S)-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4]:6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基-3-烯酰胺和(R)-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4]:6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基-3-烯酰胺
Figure PCTCN2022074328-appb-000154
将化合物乙烯乙醇酸(9.61mg，94.07μmol)用DMF(2mL)溶解，加入HATU(44.70mg,117.58μmol)、化合物SM1-1(25.00mg，0.047mmol)和DIPEA(24.30mg,188.13μmol)，25℃反应2小时。反应液经减压浓缩，浓缩物直接经制备高效液相色谱纯化，得标题化合物1-7-A(4.00mg)和1-7-B(1.38mg)。
色谱柱:SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A:乙腈；流动相B:水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 30 70 28
2 30 70 28
18 90 10 28
保留时间：1-7-A：8.7min；1-7-B：9.1min.
1-7-A结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.51(s,J＝8.8Hz,1H),7.76(s,J＝10.8Hz,1H),7.29(s,1H),6.52(s,1H),6.15–6.04(m,1H),5.58–5.49(m,1H),5.42(s,2H),5.39(s,1H),5.24–5.01(m,3H),4.54(s,J＝4.9Hz,1H),3.24–3.05(m,2H),2.37(s,3H),2.16(s,2H),1.91–1.79(m,2H),0.87(t,J＝7.2Hz,3H).
ESI-MS(m/z):520.1[M+H] +.
1-7-B结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.48(s,1H),7.78(s,1H),7.30(s,1H),6.12–5.92(m,1H),5.54–5.47(m,1H),5.42(s,2H),5.37(dt,J＝1.7Hz,1H),5.18(s,2H),5.16–5.14(m,1H),4.5–4.52(m,1H),3.22–3.08(m,2H),2.38(s,3H),2.25–2.16(m,1H),2.16–2.06(m,1H),1.93–1.79(m,2H),0.87(t,J＝7.2Hz,3H).
ESI-MS(m/z):520.1[M+H] +.
实施例三 N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基乙酰胺和N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基乙酰胺
Figure PCTCN2022074328-appb-000155
步骤一：1-氯-3-溴-2-甲基-5-硝基苯的合成
25℃下，将化合物2-1-01(5.00g,29.14mmol)溶于正庚烷(25mL)中，加入浓硫酸(25mL)，加热至50℃，50℃下分批次加入NBS(6.22g,34.97mmol)，保持50℃反应2小时，用薄层色谱检测反应(乙酸乙酯:石油醚＝1:10)，将冷却至室温的反应液滴加入冰水中，甲苯萃取，有机相合并，经亚硫酸钠溶液洗，水洗，饱和食盐水洗，无水硫酸钠干燥，减压浓缩，粗品用制备高效液相色谱纯化，制备液冷冻干燥得标题化合物4.88g。
色谱柱：C18 ODS 45mm×450mm×8.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 60 40 60
10 60 40 60
40 100 0 60
步骤二：3-氯-5-溴-4-甲基苯胺的合成
25℃下，将化合物2-1-02(4.88g,19.48mmol)溶于乙酸乙酯(100mL)中，加入铂碳(2.00g,19.48mmol,含量5％)，氢气置换后在氢气球保护下60℃反应4小时，用高效液相质谱联用色谱监测反应。将反应液过滤，滤液经浓缩，得粗品标题化合物3.68g，未经进一步纯化直接用于下一步反应。
步骤三：N-(3-氯-5-溴-4-甲基苯基)乙酰胺的合成
20℃下，将化合物2-1-03(3.63g,14.82mmol)溶于乙酸乙酯(70mL)，加入三乙胺(4.50g,44.45mmol)和醋酸酐(2.27g,22.23mmol)，保持20℃反应20小时，用高效液相质谱联用色谱监测反应。向反应液加入水，乙酸乙酯萃取，合并有机相，经无水硫酸钠干燥，减压浓缩得粗品，粗品经乙酸乙酯:石油醚＝1:5混合溶剂打浆得标题化合物2.86g。
步骤四：(Z)-4-(5-乙酰胺基-3-氯-2-甲基苯基)丁-3-烯酸的合成
20℃下，将化合物2-1-04(1.80g,6.86mmol)溶于THF(20mL)和水(5mL)中，加入乙烯基乙酸(708.31mg,8.23mmol)，DIPEA(1.95g,15.08mmol)，三(邻甲基苯基)磷(62.60mg,0.20mmol)，反应体系经氮气置换后加热至70℃反应5小时，用高效液相质谱联用色谱监测反应。向反应液中加入1N氢氧化钠溶液调节pH＝8，加入乙酸乙酯萃取。剩余水相经1N盐酸调节pH＝3，乙酸乙酯萃取，合并有机相，经无水硫酸钠干燥，减压浓缩，得标题化合物0.82g，直接用于下一步反应。
步骤五：4-(5-乙酰胺基-3-氯-2-甲基苯基)丁酸的合成
20℃下，将化合物2-1-05(2.60g,9.71mmol)溶于THF(50mL)中，加入Pd/C(0.52g，含量10％)，体系经氢气置换后在氢气球保护下40℃反应2小时，用高效液相质谱联用色谱监测反应。将反应液过滤，滤液经减压浓缩，得标题化合物2.43g，未经进一步纯化直接用于下一步反应。
步骤六：N-(3-氯-4-甲基-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将化合物2-1-06(2.43g,9.01mmol)溶于三氟乙酸(10mL)中，降温至5℃，滴加入三氟乙酸酐(3.78g,18.02mmol,2.50mL)，保持5℃反应4小时，用高效液相质谱联用色谱监测反应。将反应液加入水中，用10N氢氧化钠溶于调节pH＝9，加入乙酸乙酯萃取，合并有机相，经无水硫酸钠干燥，减压浓缩，经快速硅胶柱纯化(乙酸乙酯:石油醚＝0-20％)，得标题化合物1.53g。
步骤七:(Z)-N-(3-氯-7-(羟基亚氨基)-4-甲基-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺的合成5℃下，将叔丁醇钾(1.50g,13.37mmol)溶于THF(16mL)和叔丁醇(4mL)中，滴加入化合物2-1-07(1.53g,6.08mmol)的THF溶液(16mL)，10分钟后滴加入亚硝酸戊酯 (1.14g,9.73mmol)，保持5℃反应1小时，用高效液相质谱联用色谱监测反应。用1N盐酸将反应液调节pH＝5，乙酸乙酯萃取，合并有机相经无水硫酸钠干燥，经减压浓缩，浓缩物经甲基叔丁基醚打浆得标题化合物1.20g。
步骤八：N-(7-氨基-3-氯-4-甲基-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺
20℃下，将化合物2-1-08(0.50g,1.78mmol)溶于甲醇(8mL)和2N盐酸(8mL)中，加入Pd/C(0.15g，含量10％)，体系经氢气置换后在氢气球保护下保持5℃反应2小时，用高效液相质谱联用色谱监测反应。反应液过滤，滤液经减压浓缩，得标题化合物的盐酸盐0.52g，未经进一步纯化直接用于下一步反应。
步骤九：N,N’-(3-氯-4-甲基-8-氧代-5,6,7,8-四氢萘-1,7-二基)二乙酰胺的合成
20℃下，将化合物2-1-09(0.52g,1.70mmol)溶于吡啶(5mL)中，加入醋酸酐(2mL)，保持20℃反应2小时，用高效液相质谱联用色谱监测反应。将反应液加入水中，乙酸乙酯萃取，有机相经水洗，合并，经无水硫酸钠干燥，减压浓缩，浓缩物经快速硅胶柱纯化(乙酸乙酯:石油醚＝0-30％)得标题化合物0.22g。
步骤十：N-(8-氨基-6-氯-5-甲基-1-氧代-1,2,3,4-四氢萘-2-基)乙酰胺的合成
20℃下，将化合物2-1-10(0.45g,1.46mmol)溶于甲醇(16mL)中，加入2N盐酸(16mL)，加热至60℃反应2小时，用高效液相质谱联用色谱监测反应。向冷却后的反应液中加入饱和碳酸氢钠溶液调节pH＝8，乙酸乙酯萃取，合并有机相，经无水硫酸钠干燥，减压浓缩，得标题化合物0.23g,未经进一步纯化直接用于下一步反应。
步骤十一：N-((9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)乙酰胺的合成
将化合物2-1-11(0.23g,0.78mmol)溶于甲苯(10mL)中，加入(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃并[3,4-f]吲哚嗪-3,6,10(4H)-三酮(0.23g,0.87mmol)，对甲基苯磺酸(26.73mg,0.16mmol)，加热至140℃反应5小时，用高效液相质谱联用色谱监测反应。将反应液浓缩，粗品经快速硅胶柱纯化(甲醇:二氯甲烷＝0-10％)，得标题化合物0.15g。
步骤十二：(9S)-1-氨基-5-氯-9-乙基-9-羟基-4-甲基-1,2,3,9,12,15-六氢-10H,13H-苯并吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮的合成
将化合物2-1-12(40.00mg,0.08mmol)加入浓盐酸(1mL)中，加热至100℃反应5小时，用高效液相质谱联用色谱监测反应。将反应液过滤，滤液用制备高效液相色谱纯化，制备液经冷冻干燥得标题化合物2-23的三氟乙酸盐12.00mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％三氟乙酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 5 95 28
2 5 95 28
18 50 50 28
结构表征数据如下：
ESI-MS(m/z):452.1[M+H] +.
步骤十三:2-((叔丁基二苯基硅基)氧基)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)乙酰胺和2-((叔丁基二苯基硅基)氧基)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)乙酰胺的合成
25℃下，将化合物2-23的三氟乙酸盐(40.00mg,81.91μmol)溶于N,N-二甲基甲酰胺(1mL)中，依次加入2-((叔丁基二苯基硅基)氧基)乙酸(30.91mg,98.29μmol)，HATU(62.25mg,163.81μmol)和N,N-二异丙基乙胺(42.34mg,327.63μmol)，保持25℃反应0.5小时，用高效液相质谱联用色谱监测反应。反应完成后向反应液中加入水，用二氯甲烷/甲醇(v/v＝10/1)萃取，有机相合并，经无水硫酸钠干燥后减压浓缩，粗品经制备薄层色谱纯化(二氯甲烷:甲醇＝20:1)分离得到两个异构体，依据Rf值将两个异构体命名为2-1-13-A(15.00mg，Rf值为0.3)和2-1-13-B(12.00mg，Rf值为0.35)。
步骤十四:N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基乙酰胺和N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基乙酰胺的合成
25℃下，在两个反应瓶中分别将2-1-13-A(15.00mg)和2-1-13-B(12.00mg)溶于四氢呋喃(1mL)中，滴加入四丁基氟化铵(1M四氢呋喃溶液)/冰醋酸混合液(v/v＝13/1)(50uL)，保持25℃反应0.5小时，用高效液相质谱联用色谱监测反应。反应完成后将反应液分别用制备高效液相色谱纯化，制备液分别冻干对应得到标题化合物2-1-A(6.94mg)和2-1-B(4.00mg)。
色谱柱:SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A:乙腈；流动相B:水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 20 80 28
3 20 80 28
18 90 10 28
2-1-A结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.43(d,J＝8.8Hz,1H),8.16(s,1H),7.31(s,1H),6.55(s,1H),5.65-5.36(m,4H),5.21(q,J＝19.0Hz,2H),3.95(d,J＝5.7Hz,2H),3.26-3.11(m,2H),2.53(s,3H),2.30-2.08(m,2H),1.94-1.79(m,2H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):510.1[M+H] +.
2-1-B结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.45(d,J＝8.9Hz,1H),8.15(s,1H),7.31(s,1H),6.54(s,1H),5.64-5.35(m,4H),5.19(q,J＝19.0Hz,2H),3.97(d,J＝5.2Hz,2H),3.27-3.10(m,2H),2.51(s,3H),2.27-2.10(m,2H),1.93-1.80(m,2H),0.88(t,J＝7.3Hz,3H).
ESI-MS(m/z):510.1[M+H] +.
实施例四(2S)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基丙胺和(2S)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基丙胺
Figure PCTCN2022074328-appb-000156
步骤一:(2S)-2-((叔丁基二苯基硅基)氧基)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)丙胺和(2S)-2-((叔丁基二苯基硅基)氧基)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)丙胺的合成
25℃下，将2-23的盐酸盐(30.00mg,61.43μmol)溶于N,N-二甲基甲酰胺(1mL)中，依次加入(S)-2-((叔丁基二苯基硅)氧基)丙酸(24.21mg,73.72μmol)，HATU(35.01mg,92.14μmol)和N,N-二异丙基乙胺(23.82mg,184.29μmol)，保持25℃反应1小时，用高效液相质谱联用色谱监测反应。反应完成后向反应液中加入水，用二氯甲烷/甲醇(v/v＝10/1)萃取，有机相合并，经无水硫酸钠干燥后减压浓缩，粗品经制备薄层色谱纯化(二氯甲烷:甲醇＝15:1)分离得到两个异构体，依据Rf值将两个异构体命名为2-7-01-A(6.00mg，Rf值为0.35)和2-7-01-B(6.00mg，Rf值为0.40)。
步骤二:(2S)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基丙胺和(2S)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基丙胺的合成
25℃下，在两个反应瓶中分别将2-7-01-A(6.00mg,7.87μmol)和2-7-01-B(6.00mg,7.87μmol)溶于无水四氢呋喃(1mL)中，滴加入四丁基氟化铵(1M四氢呋喃溶液)/冰醋酸混合液(v/v＝13/1)(50μL)，保持25℃反应0.5小时，用高效液相质谱联用色谱监测反应。反应完成后将反应液分别用制备高效液相色谱纯化，制备液分别冻干对应得到标题化合 物2-7-A(2.50mg)和2-7-B(3.00mg)。
色谱柱:SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A:乙腈；流动相B:水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 15 85 28
16 90 10 28
2-7-A结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.38(d,J＝8.8Hz,1H),8.15(s,1H),7.31(s,1H),6.55(s,1H),5.56-5.47(m,2H),5.42(s,2H),5.26-5.11(m,2H),4.17-4.06(m,1H),3.27-3.10(m,2H),2.52(s,3H),2.27-2.08(m,2H),1.86(tt,J＝14.1,7.3Hz,2H),1.30(d,J＝6.7Hz,3H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):524.2[M+H] +.
2-7-B结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.48(d,J＝9.1Hz,1H),8.12(s,1H),7.30(s,1H),6.54(s,1H),5.67(d,J＝4.8Hz,1H),5.55(dd,J＝14.6,7.3Hz,1H),5.43(s,2H),5.24(d,J＝19.0Hz,1H),5.03(d,J＝19.0Hz,1H),4.21-4.08(m,1H),3.28-3.08(m,2H),2.51(s,3H),2.16(d,J＝6.2Hz,2H),1.94-1.82(m,2H),1.42(d,J＝6.8Hz,3H),0.88(t,J＝7.3Hz,3H).
ESI-MS(m/z):524.2[M+H] +.
实施例五(2S)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺和(2S)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺和(2R)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺和(2R)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺
Figure PCTCN2022074328-appb-000157
步骤一：(2S)-2-((叔丁基二苯基硅)氧基)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基乙酰胺和(2S)-2-((叔丁基二苯基硅)氧基)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基乙酰胺和(2R)-2-((叔丁基二苯基硅)氧基)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基乙酰胺和(2R)-2-((叔丁基二苯基硅)氧基)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基乙酰胺的合成
25℃下，将2-23的盐酸盐(30.00mg,61.43μmol)溶于N,N-二甲基甲酰胺(1mL)中，依次加入2-((叔丁基二苯基硅基)氧基)-2-环丙基乙酸(26.13mg,73.72μmol)，HATU(35.01mg,92.14μmol)和N,N-二异丙基乙胺(23.82mg,184.29μmol)，保持25℃反应1小时，用高效液相质谱联用色谱监测反应。反应完成后向反应液中加入水，用二氯甲烷/甲醇(v/v＝10/1)萃取，有机相合并，经无水硫酸钠干燥后减压浓缩，粗品经制备薄层色谱纯化(二氯甲烷:甲醇＝15:1)分离得到两组异构体，依据Rf值将两组异构体命名为2-12-01-A(8.00mg，Rf值为0.35)和2-12-01-B(10.00mg，Rf值为0.40)。
步骤二:(2S)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺和(2S)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺和(2R)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺和(2R)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺合成
25℃下，在两个反应瓶中分别将2-12-01-A(8.00mg,10.15μmol)和2-12-01-B(10.00mg,12.68μmol)溶于无水四氢呋喃(1mL)中，滴加入四丁基氟化铵(1M四氢呋喃溶液)/冰醋酸混合液(v/v＝13/1)(50μL),保持25℃反应0.5小时，用高效液相质谱联用色谱监测反应。反应完成后将反应液分别用制备高效液相色谱纯化，2-12-01-A为原料的反应分离出两个异构体产物，制备液分别冻干对应得到化合物2-12-A(0.77mg)、2-12-B(1.03mg)；2-12-01-B为原料的反应分离出两个异构体产物，制备液分别冻干对应得到化合物2-12-C(2.50mg)、2-12-D(1.00mg)。2-12-A/2-12-B纯化条件如下:
色谱柱:SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A:乙腈；流动相B:水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 20 80 28
2 20 80 28
18 80 20 28
出峰保留时间为：2-12-A：10.0-11.0min，2-12-B：11.0-12.5min
2-12-C/2-12-D纯化条件如下:
色谱柱:SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A:乙腈；流动相B:水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 20 80 28
2 20 80 28
18 80 20 28
出峰保留时间为：2-12-C：10.6-11.4min，2-12-D：11.4-12.5min
2-12-A结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.38(d,J＝8.9Hz,1H),8.15(s,1H),7.30(s,1H),6.54(s,1H),5.63-5.53(m,1H),5.51(d,J＝5.1Hz,1H),5.42(s,2H),5.28(d,J＝19.2Hz,1H),5.16(d,J＝19.1Hz,1H),3.60(t,J＝5.6Hz,1H),3.28-3.11(m,2H),2.52(s,3H),2.22-2.10(m,2H),1.86(tt,J＝14.1,7.3Hz,2H),1.23(d,J＝4.9Hz,1H),0.87(t,J＝7.2Hz,3H),0.56-0.36(m,4H).
ESI-MS(m/z):550.2[M+H] +.
2-12-B结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.36(d,J＝8.7Hz,1H),8.16(s,1H),7.31(s,1H),6.55(s,1H),5.57-5.48(m,1H),5.42(s,2H),5.40(d,J＝5.4Hz,1H),5.26(d,J＝19.3Hz,1H),5.18(d,J＝19.0Hz,1H),3.65-3.60(m,1H),3.26-3.12(m,2H),2.52(s,3H),2.26-2.09(m,2H),1.86(tt,J＝14.1,7.2Hz,2H),1.19-1.08(m,1H),0.87(t,J＝7.3Hz,3H),0.51-0.27(m,4H).
ESI-MS(m/z):550.2[M+H] +.
2-12-C结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.42(d,J＝9.0Hz,1H),8.15(s,1H),7.31(s,1H),6.54(s,1H),5.56(dd,J＝14.8,6.8Hz,1H),5.52(d,J＝5.2Hz,1H),5.42(s,2H),5.29(d,J＝19.2Hz,1H),5.16(d,J＝19.1Hz,1H),3.62-3.58(m,1H),3.27-3.08(m,2H),2.51(s,3H),2.27-2.08(m,2H),1.87(tt,J＝14.0,7.2Hz,2H),1.25(dd,J＝13.2,6.8Hz,1H),0.87(t,J＝7.3Hz,3H),0.64-0.28(m,4H).
ESI-MS(m/z):550.1[M+H] +.
2-12-D结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.38(d,J＝8.7Hz,1H),8.16(s,1H),7.31(s,1H),6.54(s,1H),5.59-5.49(m,1H),5.43(d,J＝5.3Hz,1H),5.43(s,2H),5.26(d,J＝19.1Hz,1H), 5.17(d,J＝19.0Hz,1H),3.64(t,J＝5.8Hz,1H),3.17(dd,J＝15.6,8.3Hz,2H),2.27-2.07(m,2H),1.94-1.79(m,2H),1.19-1.06(m,1H),0.87(t,J＝7.3Hz,3H),0.49-0.28(m,4H).
ESI-MS(m/z):550.1[M+H] +.
实施例六 N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基-2-甲基丙胺和N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基-2-甲基丙胺
Figure PCTCN2022074328-appb-000158
25℃下，将2-23的盐酸盐(30.00mg,61.43μmol)溶于N,N-二甲基甲酰胺(1mL)中，依次加入2-((叔丁基二甲基硅基)氧基)-2-甲基丙酸(16.10mg,73.72μmol)，HATU(35.01mg,92.14μmol)和N,N-二异丙基乙胺(23.82mg,184.29μmol)，保持25℃反应1小时，用高效液相质谱联用色谱监测反应。反应完成后将反应液浓缩，粗品经制备高效液相色谱纯化分离得到两个异构体，制备液分别冻干，依据出峰保留时间将两个异构体命名为2-17-A(2.65mg)和2-17-B(2.69mg)。
色谱柱:SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A:乙腈；流动相B:水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 20 80 28
2 20 80 28
18 80 20 28
出峰保留时间为：2-17-A：9.5-10.2min和2-17-B：10.4-10.6min。
2-17-A结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.36(d,J＝9.1Hz,1H),8.13(s,1H),7.30(s,1H),6.54(s,1H),5.55-5.45(m,2H),5.42(s,2H),5.28(d,J＝19.0Hz,1H),5.05(d,J＝19.0Hz,1H),3.27-3.11(m,2H),2.51(s,1H),2.24-2.10(m,2H),1.86(tt,J＝14.0,7.2Hz,2H),1.46(s,3H),1.35(s,3H),0.87(t,J＝7.3Hz,3H).
MS m/z(ESI):538.2[M+H] +.
2-17-B结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.40(d,J＝9.2Hz,1H),8.13(s,1H),7.30(s,1H),6.53 (s,1H),5.60-5.46(m,2H),5.42(s,2H),5.29(d,J＝19.0Hz,1H),5.02(d,J＝19.0Hz,1H),3.28-3.08(m,2H),2.50(s,3H),2.22-2.10(m,2H),1.87(tt,J＝14.2,7.2Hz,2H),1.47(s,3H),1.35(s,3H),0.88(t,J＝7.3Hz,3H).
MS m/z(ESI):538.2[M+H] +.
实施例七 N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-1-羟基环丙烷-1-甲酰胺和N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-1-羟基环丙烷-1-甲酰胺
Figure PCTCN2022074328-appb-000159
步骤一:1-((叔丁基二苯基硅基)氧基)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)环丙烷-1-甲酰胺和1-((叔丁基二苯基硅基)氧基)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)环丙烷-1-甲酰胺的合成
25℃下，将2-23的盐酸盐(30.00mg,61.43umol)溶于N,N-二甲基甲酰胺(1mL)中，依次加入1-((叔丁基二苯基硅基)氧基)环丙烷-1-羧酸(25.10mg,73.72umol)，HATU(35.01mg,92.14umol)和N,N-二异丙基乙胺(23.82mg,184.29μmol)，保持25℃反应1小时，用高效液相质谱联用色谱监测反应。反应完成后向反应液中加入水，用二氯甲烷/甲醇(v/v＝10/1)萃取，有机相合并，经无水硫酸钠干燥后减压浓缩，粗品经制备薄层色谱纯化(二氯甲烷:甲醇＝15:1)分离得到两个异构体，依据Rf值将两个异构体命名为2-20-01-A(4.00mg，Rf值为0.30)和2-20-01-B(4.00mg，Rf值为0.35)。
步骤二:N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-1-羟基环丙烷-1-甲酰胺和N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-1-羟基环丙烷-1-甲酰胺的合成
25℃下，在两个反应瓶中分别将2-20-01-A(4.00mg,5.17μmol)和2-20-01-B(4.00mg,5.17μmol)溶于无水四氢呋喃(1mL)中，滴加入四丁基氟化铵(1M四氢呋喃溶液)/冰醋酸混合液(v/v＝13/1)(50μL)，保持25℃反应0.5小时，用高效液相质谱联用色谱监测反应。 反应完成后将反应液分别用制备高效液相色谱纯化，制备液分别冻干对应得到标题化合物2-20-A(0.71mg)和2-20-B(1.05mg)。
2-20-A纯化条件如下:
色谱柱:SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A:乙腈；流动相B:水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 15 85 28
16 90 10 28
2-20-B纯化条件如下:
色谱柱:SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A:乙腈；流动相B:水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 20 80 28
2 20 20 28
18 80 20 28
2-20-A结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.58(d,J＝9.0Hz,1H),8.15(s,1H),7.31(s,1H),6.55(s,1H),6.30(s,1H),5.55(dd,J＝13.2,8.2Hz,1H),5.43(s,2H),5.26(d,J＝19.0Hz,1H),5.10(d,J＝19.0Hz,1H),3.29-3.09(m,2H),2.52(s,3H),2.31-2.15(m,2H),1.93-1.80(m,2H),1.25-1.14(m,2H),0.98-0.90(m,2H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):536.2[M+H] +.
2-20-B结构表征数据如下:
1H NMR(400MHz,DMSO-d 6)δ8.63(d,J＝9.0Hz,1H),8.15(s,1H),7.31(s,1H),6.54(s,1H),6.35(s,1H),5.55(dd,J＝13.5,8.6Hz,1H),5.43(s,2H),5.29(d,J＝19.1Hz,1H),5.08(d,J＝19.1Hz,1H),3.28-3.10(m,2H),2.51(s,3H),2.30-2.14(m,2H),1.93-1.81(m,2H),1.26-1.14(m,2H),1.02-0.90(m,2H),0.89(d,J＝10.9Hz,3H).
ESI-MS(m/z):536.2[M+H] +.
实施例八(1S,9S)-1-氨基-4-氯-9-乙基-5-氟-9-羟基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮和(1R,9S)-1-氨基-4-氯-9-乙基-5-氟-9-羟基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4'：6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮
Figure PCTCN2022074328-appb-000160
步骤一：3-溴-4-氯-5-氟苯胺的合成
将化合物3-1-01(2.00g,10.53mmol)溶于N,N-二甲基甲酰胺(30mL)中，然后缓慢加入N-氯代丁二酰亚胺(1.69g,12.63mmol)，加毕，室温反应16小时，用高效液相质谱联用色谱检测反应。反应液经减压浓缩得粗品，粗品经快硅胶柱纯化(乙酸乙酯:石油醚＝0-25％)得标题化合物0.95g。
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ6.77(dd,J＝2.5,1.4Hz,1H),6.51(dd,J＝11.7,2.5
Hz,1H),5.84(s,2H).
步骤二：N-(3-溴-4-氯-5-氟苯基)乙酰胺的合成
将化合物3-1-02(0.95g,4.23mmol)溶于乙酸乙酯(20mL)中，氮气保护下加入乙酸酐(648.13mg,6.35mmol)，加毕，升温至50℃反应15小时，用高效液相质谱联用色谱检测反应。反应液用甲醇(5mL)淬灭后，直接经减压蒸干得粗品，粗品经快速硅胶柱纯化(乙酸乙酯:石油醚＝0-40％)得标题化合物1.01g。
结构表征数据如下：
ESI-MS(m/z):265.9[M+H] +.
步骤三：(E)-4-(5-乙酰氨基-2-氯-3-氟苯基)-3-丁烯酸的合成
将化合物3-1-03和3-丁烯酸(387.65mg,4.50mmol)溶于1,4二氧六环(24mL)和水(8mL)的混合溶剂中，然后加入N,N-二异丙基乙胺(1.45g,11.26mmol)，三(邻甲基苯基)磷(114.21mg,375.24μmol)和醋酸钯(42.12mg,187.62μmol)，加毕，反应体系用氮气置换三次，并在氮气氛围下升温至100℃反应16小时，用高效液相质谱联用色谱检测反应。反应液冷却至室温后，加入1N的氢氧化钠水溶液(60mL)和乙酸乙酯(50mL)振荡分层。分出下层水相后，用4mol/L盐酸水溶液调节pH至3左右，然后用乙酸乙酯萃取，合并 有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，滤液经减压蒸干，得到标题化合物的粗品1.00g。
结构表征数据如下：
ESI-MS(m/z):272.0[M+H] +.
步骤四：4-(5-乙酰氨基-2-氯-3-氟苯基)丁酸的合成
将化合物3-1-04的粗品(1.00g,3.68mmol)溶于四氢呋喃(15mL)，然后加入10％钯碳(0.10g)，加毕，然后用氢气球置换反应体系三次，并在氢气氛围下反应4小时，用高效液相质谱联用色谱检测反应。将反应液过滤，滤液减压浓缩干，得标题化合物的粗品1.00g。
结构表征数据如下：
ESI-MS(m/z):274.0[M+H] +.
步骤五：N-(4-氯-3-氟-8-氧-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将化合物3-1-05的粗品(1.00g,3.65mmol)溶于三氟乙酸(5mL)中，降温至5℃后，缓慢加入三氟乙酸酐(3.84g,18.27mmol,2.54mL)，加毕，保持5℃反应2小时，用高效液相质谱联用色谱检测反应。反应液缓慢倒入水中，然后用乙酸乙酯萃取，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥有机相，然后过滤，滤液经减压蒸干得粗品，粗品经快速硅胶柱纯化，得标题化合物0.43g。
结构表征数据如下：
ESI-MS(m/z):256.1[M+H] +.
步骤六：N-(4-氯-3-氟-7-(羟基亚氨基)-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将四氢呋喃(16mL)和叔丁醇(4mL)加入反应液瓶中，冰浴降温至5℃后，加入叔丁醇钾(415.18mg,3.70mmol)，然后将化合物3-1-06(0.43mg,1.68mmol)溶于四氢呋喃(1mL)中，并缓慢滴加入反应液，10分钟后再加入亚硝酸异戊酯(315.24mg,2.69mmol)，加毕，保持5℃反应1小时，用高效液相质谱联用色谱检测反应。反应液用饱和氯化铵水溶液淬灭后，用乙酸乙酯萃取，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥有机相，然后过滤，滤液经减压浓缩，得标题化合物的粗品455.00mg。
结构表征数据如下：
ESI-MS(m/z):285.0[M+H] +.
步骤七：N-(7-氨基-4-氯-3-氟-8-氧-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将化合物3-1-07的粗品(0.40g,1.41mmol)溶于甲醇(10mL)中，然后加入3mol/L的盐酸水溶液(1mL)和10％钯碳(40.00mg)，加毕，用氢气球置换反应体系三次，并在氢气氛围下室温反应1小时，用高效液相质谱联用色谱检测反应。将反应液过滤，滤液经减压浓缩干，得标题化合物的盐酸盐粗品0.43g。
结构表征数据如下：
ESI-MS(m/z):271.0[M+H] +.
步骤八：(9H-芴-9-基)甲基(8-乙酰胺-5-氯-6-氟-1-氧代-1,2,3,4-四氢萘-2-基)氨基甲酸酯的合成
将化合物3-1-08的盐酸盐粗品(0.43g,1.19mmol)溶于1,4-二氧六环(15mL)中，然后加入碳酸氢钠(400.35mg,4.77mmol)、水(5mL)和9-芴甲基-N-琥珀酰亚胺基碳酸酯(481.81mg,1.43mmol)，加毕，室温下搅拌反应2小时，用高效液相质谱联用色谱检测反应。将反应液倒入水中，然后用乙酸乙酯萃取，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥有机相，过滤，滤液经减压浓缩得粗品。粗品经C18反相柱纯化(乙腈：0.05％甲酸水＝20％-100％)，得标题化合物301.00mg。
结构表征数据如下：
ESI-MS(m/z):493.2[M+H] +.
步骤九：(9H-芴-9-基)甲基(8-氨基-5-氯-6-氟-1-氧代-1,2,3,4-四氢萘-2-基)氨基甲酸酯的合成
将化合物3-1-09(300.00mg,608.61μmol)溶于二氧六环(5mL)中，加入12mol/L的浓盐酸(1mL)，加毕，升温至60℃反应2小时，用高效液相质谱联用色谱检测反应。将反应液倒入水中，然后用乙酸乙酯萃取，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥有机相，过滤，滤液经减压浓缩得粗品。粗品经快速硅胶柱纯化(乙酸乙酯:石油醚＝0-50％)，得标题化合物198.00mg。
结构表征数据如下：
ESI-MS(m/z):451.1[M+H] +.
步骤十：(9H-芴-9-基)甲基((9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪并[1,2-b]喹啉-1-基)氨基甲酸酯的合成
将(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃并[3,4-f]吲哚嗪-3,6,10(4H)-三酮(138.72mg,526.96μmol)和化合物3-1-10(198.00mg,439.13μmol)加入甲苯(10mL)中，然后再加入对甲苯磺酸(75.53mg,439.13μmol)，加毕，升温至140℃反应4小时，反应液直接140℃下减压蒸干得粗品，粗品经快速硅胶柱纯化(甲醇:二氯甲烷＝0-5％)，得标题化合物256.00mg。
结构表征数据如下：
ESI-MS(m/z):678.1[M+H] +.
步骤十一：(1S,9S)-1-氨基-4-氯-9-乙基-5-氟-9-羟基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4'：6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮和(1R,9S)-1-氨基-4-氯-9-乙基-5-氟-9-羟基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮的合成
将化合物3-1-11(201.18mg,296.67μmol)溶于N,N-二甲基甲酰胺(4mL)中，然后加入二乙胺(108.49mg,1.48mmol)，加毕，室温反应0.5小时，用高效液相质谱联用色谱检测反应。反应液经减压蒸出乙二胺后，用1mol/L盐酸水溶液调pH至2-3后，反应液直 接用制备高效液相色谱纯化，得标题化合物3-1-A(44.00mg)，3-1-B(43.00mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 10 90 28
3 10 90 28
18 70 30 28
3-1-A(6min LCMS出峰靠前，保留时间：1.276min)
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.00(d,J＝10.3Hz,1H),7.33(s,1H),6.54(s,1H),5.62(d,J＝19.3Hz,1H),5.44(s,2H),5.38(d,J＝19.3Hz,1H),4.43-4.38(m,1H),3.28-3.10(m,2H),2.22-2.12(m,1H),2.12-2.02(m,1H),1.93-1.80(m,2H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):456.1[M+H] +.
3-1-B(6min LCMS出峰靠后，保留时间：1.300min)结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ7.98(d,J＝10.3Hz,1H),7.32(s,1H),5.61(d,J＝19.4Hz,1H),5.44(s,2H),5.32(d,J＝19.4Hz,1H),4.44-4.36(m,1H),3.33-3.25(m,1H),3.22-3.11(m,1H),2.23-2.13(m,1H),2.11-2.03(m,1H),1.96-1.82(m,2H),0.89(t,J＝7.3Hz,3H).
ESI-MS(m/z):456.1[M+H] +.
6min LCMS条件：
色谱柱：Waters SunFire C18 OBD 4.6mm×50mm×5.0μm
流动相A：0.05％乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 90 10 2
4.2 10 90 2
5.7 10 90 2
5.71 90 10 2
6.70 90 10 2
实施例九 N-((1S,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基乙酰胺和N-((1R,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基乙酰胺
Figure PCTCN2022074328-appb-000161
步骤一：(S)-10-苄基-23-(2-(甲磺酸基)嘧啶-5-基)-6,9,12,15,18-五氧代-3-氧代-5,8,11,14,17-五氮杂十八烷-22-炔羧酸的合成
将化合物3-4-01(30.00mg,70.00μmol)溶于N,N-二甲基甲酰胺(1mL)中，加入2,5-二氧吡咯烷-1-基6-(2-(甲磺酸)嘧啶-5-基)己基-5-炔酰胺(28.00mg,77.00μmol)，室温反应1小时，用高效液相质谱联用色谱监测反应。反应液直接用制备高效液相色谱纯化，制备液冷冻干燥得标题化合物3-4-03(20.00mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 10 90 28
2.00 10 90 28
18.00 90 10 28
结构表征数据如下：
ESI-MS(m/z):691.0[M+18] +.
步骤二：N-((S)-10-苄基-1-((1S,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)氨基)-1,6,9,12,15-五氧代-3-氧杂-5,8,11,14-四氮杂十六烷-16-基)-6-(2-(甲基磺酰基)嘧啶-5-基)己基-5-酰胺和N-((S)-10-苄基-1-((1R,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧代- 2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)氨基)-1,6,9,12,15-五氧代-3-氧杂-5,8,11,14-四氮杂十六烷-16-基)-6-(2-(甲基磺酰基)嘧啶-5-基)己基-5-酰胺的合成
将单一构型的化合物3-1-A(36.00mg,79.70μmol)和化合物3-4-03(64.43mg,95.64μmol)溶于N,N-二甲基甲酰胺(2mL)中，然后加入4-(4,6-二甲氧基三嗪-2-基)-4-甲基吗啉盐酸盐(46.98mg,159.40μmol)和三乙胺(24.19mg,239.10μmol)，加毕，室温反应1小时，用高效液相质谱联用色谱检测反应。反应液直接经高效液相色谱纯化，得到单一构型的标题化合物3-4-04-A(51.00mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 30 70 28
3 30 70 28
18 90 10 28
结构表征数据如下：
ESI-MS(m/z):1111.0[M+H] +.
将单一构型的化合物3-1-B(36.00mg,79.70μmol)和化合物3-4-03(64.43mg,95.64μmol)溶于N,N-二甲基甲酰胺(2mL)中，然后加入4-(4,6-二甲氧基三嗪-2-基)-4-甲基吗啉盐酸盐(46.98mg,159.40μmol)和三乙胺(24.19mg,239.10μmol)，加毕，室温反应1小时，用高效液相质谱联用色谱检测反应。反应液直接经高效液相色谱纯化，得到单一构型的标题化合物3-4-04-B(52.00mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 30 70 28
3 30 70 28
18 90 10 28
结构表征数据如下：
ESI-MS(m/z):1111.0[M+H] +.
步骤三：N-((1S,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基乙酰胺和N-((1R,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基乙酰胺的合成
称取化合物3-4-04-A(40.00mg,35.99μmol)溶于二氯甲烷(2mL)和甲醇(1mL)的混合溶剂中，然后加入4mol/L盐酸乙酸乙酯(1mL)，加毕，室温反应0.5小时，用高效液相 质谱联用色谱检测反应。反应液直接经减压浓缩干得粗品，粗品经高效液相色谱纯化，得单一构型的标题化合物3-4-A(4.75mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 15 85 28
3 15 85 28
18 90 10 28
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.50(d,J＝8.9Hz,1H),8.05(d,J＝10.3Hz,1H),7.33(s,1H),6.55(s,1H),5.67-5.60(m,1H),5.49(t,J＝5.8Hz,1H),5.43(s,2H),5.21(s,2H),3.96(d,J＝5.8Hz,2H),3.32-3.22(m,2H),2.28-2.15(m,2H),1.93-1.80(m,2H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):514.0[M+H] +.
称取化合物3-4-04-B(40.00mg,35.99μmol)溶于二氯甲烷(2mL)和甲醇(1mL)的混合溶剂中，然后加入4mol/L盐酸乙酸乙酯(1mL)，加毕，室温反应0.5小时，用高效液相质谱联用色谱检测反应。反应液直接经减压浓缩干得粗品，粗品经高效液相色谱纯化，得单一构型的标题化合物3-4-B(8.24mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 15 85 28
3 15 85 28
18 90 10 28
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.52(d,J＝9.0Hz,1H),8.05(d,J＝10.3Hz,1H),7.34(s,1H),6.55(s,1H),5.68-5.58(m,1H),5.53(t,J＝5.8Hz,1H),5.43(d,J＝2.9Hz,2H),5.20(d,J＝7.3Hz,2H),3.97(d,J＝5.7Hz,2H),3.31-3.21(m,2H),2.26-2.15(m,2H),1.92-1.82(m,2H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):514.0[M+H] +.
实施例十(S)-N-(2-(4-乙基-8-氟-4-羟基-9-甲基-3,14-二氧代-3,4,12,14-四氢-1H-吡喃[3',4':6,7]吲哚嗪并[1,2-b]喹啉-11-基)乙基)-2-羟基-N-异丙基乙酰胺
Figure PCTCN2022074328-appb-000162
步骤一：1-(4-氟-3-甲基苯基)-3-(异丙胺基)丙烷-1-酮的合成
20℃下，将化合物4-12-01(500.00mg,3.29mmol)、甲醛水溶液(2.5mL，37％)和异丙胺(388.46mg,6.57mmol)于异丙醇(5mL)中，0℃下滴加浓盐酸(2.5mL)，反应液100℃下搅拌16小时，用高效液相质谱联用色谱检测反应。反应液经减压浓缩得粗品，粗品经制备高效液相色谱纯化，制备液冷冻干燥，得标题化合物200.00mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 10 90 28
2.00 10 90 28
18.00 90 10 28
结构表征数据如下：
ESI-MS(m/z):224.1[M+H] +.
步骤二：1-(4-氟-5-甲基-2-硝基苯基)-3-(异丙胺基)丙烷-1-酮的合成
0℃下，将化合物4-12-02(100.00mg,0.49mmol)于浓硫酸(0.5mL)中，加硝酸钾(54.34mg,0.54mmol)，反应液保持0℃反应1小时，用高效液相质谱联用色谱检测反应。将反应液倒入冰水中，经反相柱纯化(乙腈:0.05％甲酸水＝0-30％)得到标题化合物90.00mg。
结构表征数据如下：
ESI-MS(m/z):269.0[M+H] +.
步骤三：1-(2-氨基-4-氟-5-甲基苯基)-3-(异丙胺基)丙烷-1-酮的合成
25℃下，将化合物4-12-03(200.00mg,0.75mmol)于甲醇(20.0mL)中，加10％钯碳(10.00mg)，反应液氢气置换，并于氢气环境中保持20℃反应16小时，用高效液相质谱联用色谱检测反应。将反应液过滤，经减压浓缩，得标题化合物183.00mg。
结构表征数据如下：
ESI-MS(m/z):239.1[M+H] +.
步骤四：(S)-4-乙基-8-氟-4-羟基-11-(2-(异丙胺基)乙基)-9-甲基-1,12-二氢-14H-吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-3,14(4H)-二酮的合成
25℃下，将化合物4-12-04(50.00mg,0.21mmol)和(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃并[3,4-f]吲哚嗪-3,6,10(4H)-三酮(55.23mg,0.21mmol)于甲苯(3mL)中，加对甲苯磺酸(3.61mg,0.02mmol)，反应液130℃反应4小时，用高效液相质谱联用色谱检测反应。反应液经减压浓缩，粗品用制备高效液相色谱纯化，制备液冷冻干燥，得标题化合物的三氟乙酸盐2.00mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％三氟乙酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 8 92 28
2.00 8 92 28
18.00 60 40 28
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.58(s,2H),8.22(d,J＝8.1Hz,1H),7.97(d,J＝10.7Hz,1H),7.35(s,1H),6.59(s,1H),5.47(s,2H),5.41(s,2H),3.58-3.45(m,3H),3.31-3.23(m,2H),2.56(s,3H),1.98-1.80(m,2H),1.26(d,J＝6.3Hz,6H),0.89(t,J＝7.3Hz,3H).
ESI-MS(m/z):466.2[M+H] +
步骤五：(S)-2-((叔丁基二苯基甲硅烷基)氧基)-N-(2-(4-乙基-8-氟-4-羟基-9-甲基-3,14-二氧代-3,4,12,14-四氢-1H-吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-11-基)乙基)-N-异丙基乙酰胺的合成
将化合物4-12-05(22.00mg,47.26μmol)和2-((叔丁基二苯基甲硅烷基)氧基)乙酸(16.35mg,51.99μmol)溶于N,N-二甲基甲酰胺(1mL)中，然后加入HATU(21.55mg,56.71μmol)和N,N-二异丙基乙胺(18.32mg,141.78μmol)，加毕，室温反应0.5小时，用高效液相质谱联用色谱检测反应。反应液直接经C18反相柱纯化(乙腈:0.05％甲酸水溶液＝30％-100％)，得标题化合物18.00mg。
结构表征数据如下：
ESI-MS(m/z):762.3[M+H] +.
步骤六：(S)-N-(2-(4-乙基-8-氟-4-羟基-9-甲基-3,14-二氧代-3,4,12,14-四氢-1H-吡喃[3',4':6,7]吲哚嗪并[1,2-b]喹啉-11-基)乙基)-2-羟基-N-异丙基乙酰胺的合成
将化合物4-12-06(18.00mg,23.62μmol)溶于N,N-二甲基甲酰胺(1mL)中，然后加入氟化钾(6.86mg,118.12μmol)，加毕升温至50℃反应1小时，用高效液相质谱联用色谱检测反应。反应液直接经高效液相色谱纯化得标题化合物1.53mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 15 85 28
18.00 90 10 28
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.53(d,J＝8.2Hz,1H),7.91(d,J＝10.8Hz,1H),7.32(s,1H),6.55(s,1H),5.44(d,J＝13.8Hz,4H),4.72(t,J＝5.5Hz,1H),4.21(d,J＝5.5Hz,2H),3.99-3.90(m,1H),3.54-3.38(m,4H),2.54(s,3H),1.92-1.83(m,2H),1.17(dd,J＝6.6,3.1Hz,6H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):524.2[M+H] +.
实施例十一(S)-N-((4-乙基-8-氟-4-羟基-9-甲基-3,14-二氧基-3,4,12,14-四氢-1H-吡喃[3',4':6,7]吲哚嗪并[1,2-b]喹啉-11-基)甲基)-1-羟基环丙烷甲酰胺
Figure PCTCN2022074328-appb-000163
将原料(S)-11-(氨基甲基)-4-乙基-8-氟-4-羟基-9-甲基-1H-吡喃[3',4':6,7]吲哚嗪并[1,2-b]喹啉-3,14(4H,12H)-二酮(4-10-01，参照专利WO2020219287合成方法制备，30.00mg，67.00μmol)，1-羟基环丙烷羧酸(7.56mg，0.074mmol)溶于DMF(1mL)，搅拌下加入HBTU(34.30mg，0.14mmol)和二异丙基乙胺(26.09mg，0.20mmol)，室温反应4小时。加水和乙酸乙酯搅拌，静置分液，有机相用饱和食盐水洗涤后减压浓缩。浓缩物经制备薄层色谱纯化(二氯甲烷:甲醇＝20:1)，再用制备高效液相色谱纯化得固体1.20mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 20 80 28
2.00 20 80 28
18.00 80 20 28
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.96(t,J＝6.0Hz,1H),8.51(d,J＝8.0Hz,1H),7.90(d,J＝10.8Hz,1H),7.31(s,1H),6.53(s,1H),6.30(s,2H),6.30(s,1H),5.52(s,2H),5.44(s,2H),4.84(d,J＝6.0Hz,2H),2.51(s,3H),1.91-1.81(m,2H),1.01(dd,J＝7.2,4.1Hz,2H),0.87(t,J＝7.3Hz,3H),0.83(t,J＝3.6Hz,2H).
ESI-MS(m/z):494.1[M+1] +.
实施例十二(1S,9S)-1-氨基-9-乙基-5-氟-9-羟基-1,4-二甲基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮和(1R,9S)-1-氨基-9-乙基-5-氟-9-羟基-1,4-二甲基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮
Figure PCTCN2022074328-appb-000164
步骤一：(E)-4-(5-乙酰氨基-3-氟-2-甲基苯基)-2-甲基-3-丁烯酸的合成
称取N-(3-溴-5-氟-4-甲基苯基)乙酰胺(2.00g,8.13mmol)和2-甲基-3-丁烯酸(976.44mg,9.75mmol)溶于1,4-二氧六环(15mL)和水(5mL)的混合溶剂中，然后加入三(邻甲苯基)膦(247.37mg,812.76μmol)、醋酸钯(91.24mg,406.38μmol)和N,N-二异丙基乙胺(2.31g,17.88mmol)，加毕，氮气置换反应体系三次，并在氮气氛围下升温至80℃反应3小时，用高效液相质谱联用色谱检测反应。反应液冷却至室温后，加入1mol/L氢氧化钠水溶液(60mL)和乙酸乙酯(50mL)振荡分层。分出下层水相后，用4mol/L盐酸水溶液调 节pH至3左右，然后用乙酸乙酯萃取，合并有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，滤液经减压浓缩得标题化合物1.90g。
结构表征数据如下：
ESI-MS(m/z):266.1[M+H] +.
步骤二：4-(5-乙酰氨基-3-氟-2-甲基苯基)-2-甲基丁酸的合成
将(E)-4-(5-乙酰氨基-3-氟-2-甲基苯基)-2-甲基-3-丁烯酸(1.90g,7.16mmol)溶于甲醇(40mL)中，氮气保护下加入10％钯碳(0.15g)，然后用氢气球置换反应体系三次，并在氢气氛围下反应2小时，用高效液相质谱联用色谱检测反应。将反应液过滤，滤液经减压浓缩干得标题化合物1.51g。
结构表征数据如下：
ESI-MS(m/z):268.1[M+H] +.
步骤三：N-(3-氟-4,7-二甲基-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺的合成
称取4-(5-乙酰氨基-3-氟-2-甲基苯基)-2-甲基丁酸(1.50g,5.61mmol)溶于三氟乙酸(20mL)中，降温至5℃后滴加入三氟乙酸酐(2.36g,11.22mmol)，加毕，保持5℃反应2小时，用高效液相质谱联用色谱检测反应。将反应液缓慢倒入饱和碳酸氢钠水溶液中中，然后用乙酸乙酯萃取，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥有机相，过滤，滤液经减压蒸干得粗品，粗品经快速硅胶柱纯化(乙酸乙酯:石油醚＝0-30％)得标题化合物1.05g。
结构表征数据如下：
ESI-MS(m/z):250.1[M+H] +.
步骤四：N-(7-溴-3-氟-4,7-二甲基-8-氧代5,6,7,8-四氢萘-1-基)乙酰胺的合成
称取N-(3-氟-4,7-二甲基-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺(0.55g,2.21mmol)溶于醋酸(8mL)中，然后加入溴素(387.85mg,2.43mmol)，加毕，升温至50℃反应15小时，用高效液相质谱联用色谱检测反应。反应液直接经减压蒸干得粗品，粗品经快速硅胶柱纯化(乙酸乙酯:石油醚＝0-30％)得标题化合物461.00mg。
结构表征数据如下：
ESI-MS(m/z):328.0[M+H] +.
步骤五：N-(7-叠氮基-3-氟-4,7-二甲基-8-氧代5,6,7,8-四氢萘-1-基)乙酰胺的合成
称取N-(7-溴-3-氟-4,7-二甲基-8-氧代5,6,7,8-四氢萘-1-基)乙酰胺(460.00mg,1.40mmol)溶于N,N-二甲基甲酰胺(10mL)中，然后加入叠氮化钠(273.37mg,4.21mmol)，加毕，室温反应1小时，用高效液相质谱联用色谱检测反应。将反应液缓慢倒入水中，然后用乙酸乙酯萃取，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥有机相，过滤，滤液经减压蒸干得粗品，粗品经快速硅胶柱纯化(乙酸乙酯:石油醚＝0-50％)，得标题化合物347.00mg。
结构表征数据如下：
ESI-MS(m/z):291.1[M+H] +.
步骤六：N-(7-氨基-3-氟-4,7-二甲基-8-氧代5,6,7,8-四氢萘-1-基)乙酰胺的合成
称取N-(7-叠氮基-3-氟-4,7-二甲基-8-氧代5,6,7,8-四氢萘-1-基)乙酰胺(347.00mg,1.20mmol)溶于四氢呋喃(10mL)中，氮气保护下加入10％钯碳(30.00mg)，然后用氢气球置换反应体系三次，并在氢气氛围下反应2小时，用高效液相质谱联用色谱检测反应。将反应液过滤，滤液经减压浓缩干得粗品，粗品经C18反相柱纯化(乙腈:0.05％甲酸水溶液＝0％-30％)得标题化合物205.00mg。
结构表征数据如下：
ESI-MS(m/z):265.1[M+H] +.
步骤七：(9H-芴-9-基)甲基(8-乙酰氨基-6-氟-2,5-二甲基-1-氧代-1,2,3,4-四氢萘-2-基)氨基甲酸酯的合成
称取N-(7-氨基-3-氟-4,7-二甲基-8-氧代5,6,7,8-四氢萘-1-基)乙酰胺(200.00mg,756.73μmol)溶于1,4-二氧六环(6mL)和水(3mL)的混合溶剂中，然后加入碳酸氢钠(254.28mg,3.03mmol)和9-芴甲基-N-琥珀酰亚胺基碳酸酯(650.55mg,1.14mmol)，加毕，室温下搅拌反应2小时，用高效液相质谱联用色谱检测反应。将反应液缓慢倒入水中，然后用乙酸乙酯萃取，合并有机相，用饱和食盐水洗涤，经无水硫酸钠干燥有机相，过滤，滤液经减压蒸干得粗品，粗品经C18反相柱纯化(乙腈:0.05％甲酸水溶液＝20％-80％)得标题化合物301.00mg。
结构表征数据如下：
ESI-MS(m/z):487.0[M+H] +.
步骤八：(9H-芴-9-基)甲基(8-氨基-6-氟-2,5-二甲基-1-氧代1,2,3,4-四氢萘-2-基)氨基甲酸酯的合成
将(9H-芴-9-基)甲基(8-乙酰氨基-6-氟-2,5-二甲基-1-氧代-1,2,3,4-四氢萘-2-基)氨基甲酸酯(101.00mg,207.59μmol)溶于1,4-二氧六环(5mL)中，然后加入3mol/L盐酸水溶液(5mL)，加毕，升温至50℃反应15小时，用高效液相质谱联用色谱检测反应。将反应液缓慢倒入饱和碳酸氢钠水溶液中，然后用乙酸乙酯萃取，合并有机相用饱和食盐水洗涤，无水硫酸钠干燥有机相，过滤，滤液经减压蒸干得粗品，粗品经快速硅胶柱纯化(甲醇:二氯甲烷＝0％-5％)得标题化合物71.00mg。
结构表征数据如下：
ESI-MS(m/z):445.2[M+H] +.
步骤九：(9H-芴-9-基)甲基((9S)-9-乙基-5-氟-9-羟基-1,4-二甲基-10,13-二氧代-2,3,9,10,13,15–六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)氨基甲酸酯的合成
将(9H-芴-9-基)甲基(8-氨基-6-氟-2,5-二甲基-1-氧代1,2,3,4-四氢萘-2-基)氨基甲酸酯(35.00mg,132.96μmol)和(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃并[3,4-f]吲哚嗪-3,6,10(4H)- 三酮(49.25mg,110.80μmol)加入甲苯(3mL)中，然后加入对甲苯磺酸(19.08mg,110.80μmol)，加毕，升温至140℃反应4小时，反应液直接140℃下经减压蒸干得粗品。粗品经C18反相柱纯化(乙腈:0.05％甲酸水溶液＝20％-80％)得标题化合物21.00mg。
结构表征数据如下：
ESI-MS(m/z):672.2[M+H] +.
步骤十：(1S,9S)-1-氨基-9-乙基-5-氟-9-羟基-1,4-二甲基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮和(1R,9S)-1-氨基-9-乙基-5-氟-9-羟基-1,4-二甲基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮的合成
将(9H-芴-9-基)甲基((9S)-9-乙基-5-氟-9-羟基-1,4-二甲基-10,13-二氧代-2,3,9,10,13,15–六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)氨基甲酸酯(21.00mg,31.26μmol)溶于N,N-二甲基甲酰胺(1mL)中，然后加入二乙胺(0.2mL)，加毕，室温反应0.5小时，用高效液相质谱联用色谱检测反应。反应液经减压蒸出乙二胺后，用1mol/L盐酸水溶液调节pH至2-3后，反应液直接用制备高效液相色谱纯化，得两个异构体5-13-A(1.30mg)和5-13-B(1.68mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 10 90 28
3.00 10 90 28
18.00 90 10 28
5-13-A(6min LCMS出峰靠前，保留时间：1.373min)结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ7.88(d,J＝10.6Hz,1H),7.36(s,1H),6.58(s,1H),5.60(d,J＝3.5Hz,2H),5.46(d,J＝2.5Hz,2H),3.25-3.17(m,2H),2.41(s,3H),2.38-2.28(m,2H),1.91-1.84(m,2H),1.79(s,3H),0.88(t,J＝7.3Hz,3H).
ESI-MS(m/z):450.2[M+H] +.
5-13-B(6min LCMS出峰靠后，保留时间：1.523min)结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ7.76(d,J＝10.8Hz,1H),7.30(s,1H),6.52(s,1H),5.73(d,J＝19.8Hz,1H),5.50-5.40(m,3H),3.26-3.17(m,1H),3.08-2.96(m,1H),2.38(s,3H),2.19-2.11(m,1H),2.04(td,J＝13.0,5.1Hz,1H),1.91-1.79(m,2H),1.34(s,3H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):450.2[M+H] +.
6min LCMS条件：
色谱柱：Waters SunFire C18 OBD 4.6mm×50mm×5.0μm
流动相A：0.05％乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 90 10 2
4.2 10 90 2
5.7 10 90 2
5.71 90 10 2
6.70 90 10 2
实施例十三(1S,9S)-1-(氨基甲基)-9-乙基-5-氟-9-羟基-4-甲基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮和(1R,9S)-1-(氨基甲基)-9-乙基-5-氟-9-羟基-4-甲基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮
Figure PCTCN2022074328-appb-000165
步骤一：N-(7-((二甲氨基)亚甲基)-3-氟-4-甲基-8-氧基-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将化合物5-13-04(1.00g,4.25mmol)溶于N,N-二甲基甲酰胺二甲基缩醛(10mL)中，然后升温至120℃反应3小时，用高效液相质谱联用色谱检测反应。反应液冷却至室温后，直接经减压蒸干得粗品，粗品经快速硅胶柱纯化(乙酸乙酯:石油醚＝20％-100％)得标题化合物891.00mg。
结构表征数据如下：
ESI-MS(m/z):291.1[M+H] +.
步骤二：N-(7-(氨基亚甲基)-3-氟-4-甲基-8-氧-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将化合物5-7-01(0.89g,3.07mmol)溶于乙醇(25mL)中，然后加入乙酸铵(2.36g,30.65mmol)，加毕，室温反应16小时，用高效液相质谱联用色谱检测反应。将反应液溶剂减压蒸干，然后加入二氯甲烷(30mL)和水(20mL)后，搅拌静置分出有机相，用无水硫酸钠干燥，过滤，滤液经减压蒸干得标题化合物785.00mg。
结构表征数据如下：
ESI-MS(m/z):263.1[M+H] +.
步骤三：N-(7-(氨基甲基)-3-氟-4-甲基-8-氧-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将化合物5-7-02(0.80g,3.05mmol)溶于乙醇(200mL)中，然后加入10％钯碳(0.40mg)和浓盐酸(0.2mL)，加毕，用氢气球置换反应体系三次，然后在氢气氛围下室温反应3小时，用高效液相质谱联用色谱检测反应。将反应液直接过滤，滤液经减压蒸干得标题化合物的盐酸盐905.00mg。
结构表征数据如下：
ESI-MS(m/z):265.1[M+H] +.
步骤四：(9H-芴-9-基)甲基((8-乙酰胺-6-氟-5-甲基-1-氧代-1,2,3,4-四氢萘-2-基)甲基)氨基甲酸酯的合成
将化合物5-7-03的盐酸盐(0.90g,2.99mmol)溶于1,4-二氧六环(20mL)中，然后加入碳酸氢钠(1.01g,11.97mmol)、水(10mL)和9-芴甲基-N-琥珀酰亚胺基碳酸酯(1.21g,3.59mmol)，加毕，室温下搅拌反应1小时，用高效液相质谱联用色谱检测反应。反应液倒入水中，然后用乙酸乙酯萃取，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥有机相，过滤，滤液经减压整干得粗品。粗品经快速硅胶柱纯化(乙酸乙酯:石油醚＝0-50％)得标题化合物1.30g。
结构表征数据如下：
ESI-MS(m/z):487.1[M+H] +.
步骤五：(9H-芴-9-基)甲基((8-氨基-6-氟-5-甲基-1-氧代-1,2,3,4-四氢萘-2-基)甲基)氨基甲酸酯的合成
将化合物5-7-04(0.80g,1.64mmol)溶于1,4-二氧六环(20mL)中，氮气保护下加入3mol/L盐酸水溶液(20mL)，加毕，升温至60℃反应15小时，用高效液相质谱联用色谱检测反应。将反应液缓慢倒入水中，然后用乙酸乙酯萃取，合并有机相用饱和食盐水洗涤，无水硫酸钠干燥有机相，过滤，滤液经减压蒸干得粗品。粗品经快速硅胶柱纯化(乙酸乙酯:石油醚＝0-40％)得标题化合物561.00mg。
结构表征数据如下：
ESI-MS(m/z):445.1[M+H] +.
步骤六：(9H-芴-9-基)甲基(((9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)甲基)氨基甲酸酯的合成
将(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃并[3,4-f]吲哚嗪-3,6,10(4H)-三酮(597.00mg,2.27mmol)和化合物5-7-05(840.00mg,1.89mmol)加入甲苯(60mL)中，然后再加入对甲苯磺酸(325.00mg,1.89mmol)，加毕，升温至140℃反应4小时，然后反应液直接在140℃下减压蒸干得粗品。粗品经快速硅胶柱纯化(甲醇:二氯甲烷＝0-5％)得标题化合物563.00mg。
结构表征数据如下：
ESI-MS(m/z):672.2[M+H] +.
步骤七：(1S,9S)-1-(氨基甲基)-9-乙基-5-氟-9-羟基-4-甲基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮和(1R,9S)-1-(氨基甲基)-9-乙基-5-氟-9-羟基-4-甲基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮的合成
将化合物5-7-06(454.00mg,675.89μmol)溶于N,N-二甲基甲酰胺(5mL)中，然后加入二乙胺(1mL)，加毕室温反应0.5小时，用高效液相质谱联用色谱检测反应。反应液减压蒸出乙二胺后，用甲酸调节pH至2-3后，反应液直接用制备高效液相色谱纯化，制备液分别冻干对应得到标题化合物5-7-A(32.00mg)和5-7-B(56.00mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 10 90 28
3 10 90 28
18 90 10 28
5-7-A(6min LCMS出峰靠前，保留时间：1.488min)结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ7.88(d,J＝10.6Hz,1H),7.36(s,1H),6.58(s,1H),5.60(d,J＝3.5Hz,2H),5.46(d,J＝2.5Hz,2H),3.25-3.17(m,2H),2.41(s,3H),2.38-2.28(m,2H),1.91-1.84(m,2H),1.79(s,3H),0.88(t,J＝7.3Hz,3H).
ESI-MS(m/z):450.2[M+H] +.
5-7-B(6min LCMS出峰靠后，保留时间：1.596min)结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ7.76(d,J＝10.8Hz,1H),7.30(s,1H),6.52(s,1H),5.73(d,J＝19.8Hz,1H),5.50-5.40(m,3H),3.26-3.17(m,1H),3.08-2.96(m,1H),2.38(s,3H),2.19-2.11(m,1H),2.04(td,J＝13.0,5.1Hz,1H),1.91-1.79(m,2H),1.34(s,3H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):450.2[M+H] +.
6min LCMS条件：
色谱柱：Waters SunFire C18 OBD 4.6mm×50mm×5.0μm
流动相A：0.05％乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 90 10 2
4.2 10 90 2
5.7 10 90 2
5.71 90 10 2
6.70 90 10 2
实施例十四 N-((1S,9S)-9-乙基-5-氟-9-羟基-1,4-二甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-1-羟基环丙烷-1-羧酰胺或N-((1R,9S)-9-乙基-5-氟-9-羟基-1,4-二甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-1-羟基环丙烷-1-羧酰胺
Figure PCTCN2022074328-appb-000166
步骤一：1-((叔丁基二苯基甲硅烷基)氧基)-N-((1S,9S)-9-乙基-5-氟-9-羟基-1,4-二甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)环丙烷-1-甲酰胺或1-((叔丁基二苯基甲硅烷基)氧基)-N-((1R,9S)-9-乙基-5-氟-9-羟基-1,4-二甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)环丙烷-1-甲酰胺的合成
将单一构型的化合物5-13-A(10.00mg,22.25μmol)和1-((叔丁基二苯基甲硅烷基)氧基)环丙烷-1-羧酸(11.36mg,33.37μmol)溶于N,N-二甲基甲酰胺(1mL)中，然后加入HATU(12.68mg,33.37μmol)和N,N-二异丙基乙胺(8.63mg,66.74μmol)，加毕，室温反应0.5小时，用高效液相质谱联用色谱检测反应。反应液直接经C18反相柱纯化(乙腈:0.05％甲酸水溶液＝30％-100％)得到单一标题化合物5-16-01-A(7mg)。
结构表征数据如下：
ESI-MS(m/z):772.3[M+H] +.
步骤二：N-((1S,9S)-9-乙基-5-氟-9-羟基-1,4-二甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-1-羟基环丙烷-1-羧酰胺或N-((1R,9S)-9-乙基-5-氟-9-羟基-1,4-二甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-1-羟基环丙烷-1-羧酰胺的合成
将化合物5-16-01-A(7.00mg,9.07μmol)溶于N,N-二甲基甲酰胺(1mL)中，然后加入氟化钾(2.63mg,45.34μmol)，加毕升温至50℃反应1小时，用高效液相质谱联用色谱检测反应。反应液直接经高效液相色谱纯化得单一标题化合物5-16-A(1.73mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 10 90 28
18.00 90 10 28
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.26(s,1H),7.78(d,J＝10.8Hz,1H),7.30(s,1H),6.56(s,1H),6.52(s,1H),5.52(d,J＝19.3Hz,1H),5.43(d,J＝4.4Hz,2H),4.94(d,J＝19.2Hz,1H),3.30-3.24(m,1H),3.11-3.00(m,1H),2.95-2.84(m,1H),2.39(s,3H),1.98-1.80(m,3H),1.62(s,3H),0.87(t,J＝7.9Hz,3H).
ESI-MS(m/z):534.2[M+H] +.
实施例十五 N-((10S)-10-苄基-1-(((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4'；6,7]吲哚嗪并[1,2-b]喹啉-1-基)氨基)-1,6,9,12,15-五氧代-3-氧代-5,8,11,14-四氮杂十六烷-16-基)-6-(2-(甲磺酰基)嘧啶-5-基)十六烷酰胺或N-((10S)-10-苄基-1-(((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4'；6,7]吲哚嗪并[1,2-b]喹啉-1-基)氨基)-1,6,9,12,15-五氧代-3-氧代-5,8,11,14-四氮杂十六烷-16-基)-6-(2-(甲磺酰基)嘧啶-5-基)十六烷酰胺
Figure PCTCN2022074328-appb-000167
步骤一:(9S)-1-氨基-5-氯-9-乙基-9-羟基-4-甲基-1,2,3,9,12,15-六氢-10H,13H-苯并吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-10,13-二酮的分离纯化
将化合物2-23(16.00mg)用制备高效液相色谱纯化，在下述纯化条件下分离得到两个非对映异构体，得到2-23-A的三氟乙酸盐5.10mg(保留时间9.85min)和2-23-B的三氟乙酸盐7.12mg(保留时间10.62min)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％三氟乙酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 5 95 28
2 5 95 28
18 50 50 28
结构表征数据如下：
2-23-A：
1H NMR(400MHz,DMSO-d 6)δ8.42(s,3H),8.27(s,1H),7.36(s,1H),6.59(s,1H),5.78-5.63(m,1H),5.50-5.36(m,3H),5.10-5.06(m,1H),3.20-3.04(m,2H),2.56(s,3H),2.26-2.13(m,2H),1.93-1.79(m,2H),0.88(t,J＝7.2Hz,3H).
ESI-MS(m/z):452.1[M+H] +.
2-23-B：
1H NMR(400MHz,DMSO-d 6)δ8.42(s,3H),8.27(s,1H),7.36(s,1H),6.58(s,1H),5.78-5.63(m,1H),5.50-5.36(m,3H),5.10-5.06(m,1H),3.20-3.04(m,2H),2.55(s,3H),2.26-2.13(m,2H),1.93-1.79(m,2H),0.88(t,J＝7.2Hz,3H).
ESI-MS(m/z):452.0[M+H] +.
步骤二：N-((10S)-10-苄基-1-(((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4'；6,7]吲哚嗪并[1,2-b]喹啉-1-基)氨基)-1,6,9,12,15-五氧代-3-氧代-5,8,11,14-四氮杂十六烷-16-基)-6-(2-(甲磺酰基)嘧啶-5-基)十六烷酰胺或N-((10S)-10-苄基-1-(((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧代-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4'；6,7]吲哚嗪并[1,2-b]喹啉-1-基)氨基)-1,6,9,12,15-五氧代-3-氧代-5,8,11,14-四氮杂十六烷-16-基)-6-(2-(甲磺酰基)嘧啶-5-基)十六烷酰胺的合成
25℃下，将2-23-A的三氟乙酸盐(34.71mg,61.43μmol)溶于N,N-二甲基甲酰胺(1mL)中，依次加入3-4-03(49.66mg,73.72μmol)、HATU(35.01mg,92.14μmol)和N,N-二异丙基乙胺(23.82mg,184.29μmol)，保持25℃反应0.5小时,用高效液相质谱联用色谱监测反应,反应完成将反应液用制备高效液相色谱纯化(条件如下)，制备液冷冻干燥得标题化合物D-L-15 11.04mg，保留时间7.5min。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 30 70 28
3 30 70 28
18 90 10 28
结构表征数据如下：
D-L-15：
ESI-MS(m/z):1107.3[M+H] +.
实施例十六 N-((9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺
Figure PCTCN2022074328-appb-000168
步骤一：N-((9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-环丙基-2-羟基乙酰胺的合成
将单一构型的化合物3-1-A的甲酸盐(50mg,109.68μmol)和2-环丙基-2-羟基乙酸(25.47mg,219.36μmol)溶于N,N-二甲基甲酰胺(2mL)中，然后加入HATU(7.57mg,219.36μmol)和N,N-二异丙基乙胺(42.53mg,329.04μmol)，加毕，室温反应0.5小时；用高效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得标题化合物的两个异构体(3-12-A：12.96mg，3-12-B：13.56mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 30 90 28
3.00 30 90 28
18.00 90 10 28
3-12-A(6min LCMS出峰靠前)结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.44(d,J＝9.0Hz,1H),8.05(d,J＝10.2Hz,1H),7.33(s,1H),6.54(s,1H),5.62(q,J＝6.7Hz,1H),5.52(d,J＝5.1Hz,1H),5.42(s,2H),5.24(q,J＝19.2Hz,2H),3.61(dd,J＝6.2,5.1Hz,1H),3.32–3.21(m,2H),2.19(q,J＝6.5Hz,2H),1.92–1.80(m,2H),1.26–1.20(m,1H),0.87(t,J＝7.3Hz,3H),0.57–0.34(m,4H).
ESI-MS(m/z):554.0[M+H]+.
3-12-B(6min LCMS出峰靠后)结构表征数据如下：
1H NMR(400MHz,DMSO-d6)δ8.43(d,J＝8.7Hz,1H),8.06(d,J＝10.3Hz,1H),7.33(s,1H),5.57(q,J＝6.7Hz,1H),5.43(s,2H),5.30–5.17(m,2H),3.64(d,J＝6.2Hz,1H),3.29(q,J＝6.7Hz,2H),2.28–2.13(m,2H),1.93–1.78(m,2H),1.18–1.08(m,1H),0.87(t,J＝7.3Hz,3H),0.50–0.29(m,4H).
ESI-MS(m/z):554.0[M+H]+.
将另一单一构型的化合物3-1-B的甲酸盐(50mg,109.68μmol)和2-环丙基-2-羟基乙 酸(25.47mg,219.36μmol)溶于N,N-二甲基甲酰胺(2mL)中，然后加入HATU(7.57mg,219.36μmol)和N,N-二异丙基乙胺(42.53mg,329.04μmol)，加毕，室温反应0.5小时；用高效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得标题化合物的两个异构体(3-12-C：20.19mg，3-12-D：18.33mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 30 90 28
3.00 30 90 28
18.00 90 10 28
3-12-C(6min LCMS出峰靠前)结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝9.0Hz,1H),8.06(d,J＝10.3Hz,1H),7.33(s,1H),6.54(s,1H),5.62(q,J＝6.5Hz,1H),5.53(d,J＝5.1Hz,1H),5.43(s,2H),5.32–5.16(m,2H),3.61(dd,J＝6.3,5.1Hz,1H),3.32–3.22(m,2H),2.19(q,J＝6.5Hz,2H),1.92–1.80(m,2H),1.28–1.20(m,1H),0.87(t,J＝7.3Hz,3H),0.54–0.35(m,4H).
ESI-MS(m/z):554.0[M+H]+.
3-12-D(6min LCMS出峰靠后)结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.44(d,J＝8.8Hz,1H),8.05(d,J＝10.2Hz,1H),7.34(s,1H),6.55(s,1H),5.58(q,J＝6.7Hz,1H),5.45(d,J＝5.2Hz,1H),5.43(s,2H),5.31–5.14(m,2H),3.65(t,J＝5.7Hz,1H),3.33–3.21(m,2H),2.28–2.13(m,2H),1.95–1.80(m,2H),1.16–1.09(m,1H),0.88(t,J＝7.3Hz,3H),0.46–0.31(m,4H).
ESI-MS(m/z):554.0[M+H] +.
实施例十七：(S)-N-((1S,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-1-基)-2-羟基丙胺和(S)-N-((1R,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙胺的制备
Figure PCTCN2022074328-appb-000169
25℃下，将(9S)-1-氨基-4-氯-9-乙基-5-氟-9-羟基-2,3,12,15-四氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-10,13(1H,9H)-二酮(80.0mg,175.5μmol)和L-乳酸(31.6mg,351.0 μmol,)溶于DMF(3mL)中，然后加入HATU(121.1mg,351.0μmol)和DIPEA(68.0mg,526.5μmol)，室温反应2hr；反应液直接用制备高效液相色谱纯化得到化合物3-7-A(6.1mg，收率12％)和化合物3-7-B(9.6mg，收率20％)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 30 70 24
2.00 30 70 24
18.00 90 10 24
化合物3-7-A(6min LC-MS出峰靠前，保留时间2.49min)结构表征数据如下：
MS m/z(ESI):528.2[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.55(d,J＝9.2Hz,1H),8.06(d,J＝10.4Hz,1H),7.33(s,1H),6.55(s,1H),6.67(d,J＝4.8Hz,1H),5.65–5.59(m,1H),5.43(s,2H),5.29–5.21(m,1H),5.14–5.10(m,1H),4.15–4.10(m,,1H),3.27–3.20(m,1H),2.22–2.15(m,2H),1.92–1.81(m,2H),1.41(d,J＝6.8Hz,3H),1.30–1.23(m,1H),0.89–0.85(t,J＝7.2Hz,3H).
化合物3-7-B(6min LC-MS出峰靠后，保留时间2.50min)结构表征数据如下：
MS m/z(ESI):528.2[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.46(d,J＝8.8Hz,1H),8.06(d,J＝10.4Hz,1H),7.33(s,1H),6.56(s,1H),5.60–5.53(ms,1H),5.51(d,J＝5.2Hz,1H),5.43(s,2H),5.27–5.14(m,2H),4.16–4.08(m,1H),3.28–3.22(m,1H),2.22–2.19(m,2H),1.92–1.81(m,2H),1.49–1.39(m,1H),1.29(d,J＝6.8Hz,3H),0.87(t,J＝7.2Hz,3H).
实施例十八 N-((1S,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧代-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基-1-羟基环丙基甲酰胺化合物和N-((1R,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧代-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基-1-羟基环丙基甲酰胺化合物的合成
Figure PCTCN2022074328-appb-000170
25℃下，将(9S)-1-氨基-4-氯-9-乙基-5-氟-9-羟基-2,3,12,15-四氢苯并[de]吡喃并[3'，4'：6,7]吲哚嗪[1,2-b]喹啉-10,13(1H,9H)-二酮(80mg,175.49μmol)和1-羟基环丙烷羧酸(35.83mg,350.98μmol,)溶于DMF(2mL)中，然后加入HATU(121.14mg,350.98μmol)和DIPEA(68.04mg,526.47μmol)，加完后，室温反应0.5hr；反应液浓缩至干直接用高 效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得标题化合物3-17-A 5.3mg和3-17-B 3.5mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 15 85 28
2.00 15 85 28
18.00 90 10 28
结构表征数据如下：
3-17-A(6min LCMS出峰靠前，保留时间：2.657min)结构表征数据如下：
ESI-MS(m/z):540.0[M+H]+.
1H NMR(400MHz,DMSO-d 6)δ8.69(d,J＝8.8Hz,1H),8.06(d,J＝10.2Hz,1H),7.35(s,1H),6.57(s,1H),6.32(s,1H),5.62(s,1H),5.45(s,2H),5.34–5.24(m,1H),5.20–5.10(m,1H),2.26(s,2H),2.00(s,1H),1.88(s,2H),1.47–1.12(m,8H),1.01–0.80(m,6H).
3-17-A(6min LCMS出峰靠后，保留时间：2.724min)结构表征数据如下：
ESI-MS(m/z):540.0[M+H]+.
1H NMR(400MHz,DMSO-d 6)δ8.69(d,J＝8.8Hz,1H),8.06(d,J＝10.2Hz,1H),7.35(s,1H),6.57(s,1H),6.32(s,1H),5.62(s,1H),5.45(s,2H),5.34–5.24(m,1H),5.20–5.10(m,1H),2.26(s,2H),2.00(s,1H),1.88(s,2H),1.47–1.12(m,8H),1.01–0.80(m,6H).
实施例十九 N-((1S,9S)-4-环丙基-9-乙基-5-氟-9-羟基-10,13-二氧-1,2,3,9,10,12,13,15-四氢苯并[de]吡喃并[3'，4'：6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基乙酰胺化合物的合成
Figure PCTCN2022074328-appb-000171
步骤一：N-(4-氯-3-氟-7-(羟基亚氨基)-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将N-(4-氯-3-氟-8-氧-5,6,7,8-四氢萘-1-基团)乙酰胺(570mg,2.23mmol)和环丙基硼酸(574.56mg,6.69mmol)溶于1,4-二氧六环中，加入二氯二叔丁基-(4-二甲基氨基苯基)膦钯(II)(480mg,677.97μmol)和碳酸铯(2.17g,6.69mmol)，氮气保护后，微波115℃反应2h；用高效液相质谱联用色谱检测反应；反应液用乙酸乙酯稀释后过滤，滤液用乙酸乙酯(30ml*3)萃取，合并有机相，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥有机相，然 后过滤，滤液减压蒸干，粗品经柱层析硅胶柱纯化(PE：EA＝1:4)，得到标题化合物550mg。
结构表征数据如下：
ESI-MS(m/z):262.1[M+H] +.
步骤二：N-(4-环丙基-3-氟-7-(羟基亚氨基)-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将四氢呋喃(30mL)和叔丁醇(10mL)加入反应液瓶中，冰浴降温至5℃后，加入叔丁醇钾(945mg,8.42mmol)，然后将N-(4-氯-3-氟-7-(羟基亚氨基)-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺(1.0g,3.83mmol)溶于四氢呋喃(1mL)中，并缓慢滴加入反应液，10分钟后再加入亚硝酸异戊酯(718mg,6.12mmol)，加毕，保持5℃反应1小时；用高效液相质谱联用色谱检测反应；反应液用饱和氯化铵水溶液(50mL)淬灭后，用乙酸乙酯(40ml*3)萃取，合并有机相，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥有机相，然后过滤，滤液减压蒸干得标题化合物的粗品1.2g。
结构表征数据如下：
ESI-MS(m/z):291.1[M+H] +.
步骤三：N-(7-氨基-4-环丙基-3-氟-8-氧-5,6,7,8-四氢萘-1-基)乙酰胺盐酸盐的合成
将N-(4-环丙基-3-氟-7-(羟基亚氨基)-8-氧代-5,6,7,8-四氢萘-1-基)乙酰胺的粗品(1.2g,1.41mmol)溶于甲醇(7.5mL)和四氢呋喃(7.5mL)中，然后加入1mol/L的盐酸水溶液(7.5mL)和10％钯碳(450mg)，加毕，用氢气球置换反应体系三次，并在氢气氛围下室温反应1小时；用高效液相质谱联用色谱检测反应；反应液过滤，滤液减压浓缩干得到粗品1.05g。
结构表征数据如下：
ESI-MS(m/z):277.1[M+H] +.
步骤四：(9H-芴-9-基)甲基(8-乙酰胺-5-环丙基-6-氟-1-氧代-1,2,3,4-四氢萘-2-基)氨基甲酸酯的合成
将N-(7-氨基-4-环丙基-3-氟-8-氧-5,6,7,8-四氢萘-1-基)乙酰胺的盐酸盐粗品(1.05g,3.80mmol)溶于1,4-二氧六环(10mL)中，然后加入碳酸氢钠(1.3g,15.20mmol)、水(10mL)和9-芴甲基-N-琥珀酰亚胺基碳酸酯(1.54g,4.56mmol)，加毕，室温下搅拌反应2小时；用高效液相质谱联用色谱检测反应；反应液倒入水(50mL)中，然后用乙酸乙酯(40mL*3)萃取，合并有机相，用饱和食盐水(50mL)洗涤，无水硫酸钠干燥有机相，过滤，滤液减压整干得粗品。粗品经C18反相柱纯化得标题化合物2.0g。
结构表征数据如下：
ESI-MS(m/z):499.2[M+H] +.
步骤五：(9H-芴-9-基)甲基(8-氨基-5-环丙基-6-氟-1-氧代-1,2,3,4-四氢萘-2-基)氨基甲酸酯的合成
将(9H-芴-9-基)甲基(8-乙酰胺-5-环丙基-6-氟-1-氧代-1,2,3,4-四氢萘-2-基)氨基甲酸酯 (2.0g,3.21mmol，80％)溶于二氧六环(20mL)中，加入12mol/L的浓盐酸(5mL)，加毕，升温至70℃反应2小时；用高效液相质谱联用色谱检测反应；反应液倒入水(40mL)中，然后用乙酸乙酯(30mL*3)萃取，合并有机相，用饱和食盐水(40mL)洗涤，无水硫酸钠干燥有机相，过滤，滤液减压整干得粗品。粗品经柱层析硅胶柱纯化(PE：EA＝2:1)，得标题化合物740mg。
结构表征数据如下：
ESI-MS(m/z):457.3[M+H] +.
步骤六：(9H-芴-9-基)甲基((9S)-4-环丙基-9-乙基-5-氟-9-羟基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)氨基甲酸酯的合成
将(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃并[3,4-f]吲哚嗪-3,6,10(4H)-三酮(442mg,1.68mmol)和(9H-芴-9-基)甲基(8-氨基-5-环丙基-6-氟-1-氧代-1,2,3,4-四氢萘-2-基)氨基甲酸酯(640mg,1.40mmol)加入甲苯(30mL)中，然后再加人对甲苯磺酸(242mg,1.40mmol)，加毕，升温至135℃反应2小时，反应液直接140℃下减压蒸干得粗品；粗品经柱层析硅胶柱纯化(DCM：MeOH＝33:1)，得标题化合物1.02g。
结构表征数据如下：
ESI-MS(m/z):684.1[M+H] +.
步骤七：(1S,9S)-1-氨基-4-环丙基-9-乙基-5-氟-9-羟基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4'：6,7]吲哚嗪[1,2-b]喹啉-10,13-二酮(化合物5-29-1)&(1R,9S)-1-氨基-4-环丙基-9-乙基-5-氟-9-羟基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-10,13-二酮的合成
将(9H-芴-9-基)甲基((9S)-4-环丙基-9-乙基-5-氟-9-羟基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)氨基甲酸酯(1.02g,1.49mmol)溶于N,N-二甲基甲酰胺(15mL)中，然后加入二乙胺(5ml)，加毕，室温反应0.5小时；用高效液相质谱联用色谱检测反应；反应液减压蒸出乙二胺后，用1mol/L盐酸水溶液调pH至2-3后，反应液直接用制备高效液相色谱纯化得标题化合物的两个异构体(5-22-7-A：60mg；5-22-7-B：55mg)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 10 90 28
3.00 10 90 28
18.00 70 30 28
5-22-7-A(6min LCMS出峰靠前，保留时间：2.28min)结构表征数据如下：
ESI-MS(m/z):462.2[M+H] +.
5-22-7-B(6min LCMS出峰靠后，保留时间：2.35min)结构表征数据如下：
ESI-MS(m/z):462.2[M+H] +.
步骤八N-((1S,9S)-4-环丙基-9-乙基-5-氟-9-羟基-10,13-二氧-1,2,3,9,10,12,13,15-四氢苯并[de]吡喃并[3'，4'：6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基乙酰胺和N-((1R,9S)-4-环丙基-9-乙基-5-氟-9-羟基-10,13-二氧-1,2,3,9,10,12,13,15-四氢苯并[de]吡喃并[3'，4'：6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基乙酰胺的合成
25℃下，将单一构型的化合物5-28-7-A(40mg,86μmol)和乙醇酸(8mg,104μmol,)溶于DMF(2mL)中，然后加入HATU(40mg,104μmol)和DIPEA(36mg,258μmol)，加完后，室温反应0.5hr；反应液浓缩至干直接用高效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得化合物5-22-A 12.5mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 10 90 28
2.00 10 90 28
18.00 90 10 28
结构表征数据如下：
5-22-A(6min LCMS出峰靠前，保留时间：2.540min)结构表征数据如下：
ESI-MS(m/z):520.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.46(d,J＝8.9Hz,1H),7.74(d,J＝11.9Hz,1H),7.30(s,1H),6.53(s,1H),5.64–5.56(m,1H),5.49(t,J＝5.8Hz,1H),5.42(s,2H),5.19(s,2H),3.96(d,J＝5.7Hz,2H),2.25–2.10(m,2H),2.04–1.79(m,4H),1.23(s,2H),1.15–1.05(m,2H),0.87(t,J＝7.2Hz,3H),0.80–0.70(m,2H).
25℃下，将将单一构型的化合物5-28-7-B(30mg,65μmol)和乙醇酸(6mg,78μmol)溶于DMF(2mL)中，然后加入HATU(40mg,104μmol)和DIPEA(17mg,130μmol)，加完后，室温反应0.5hr；反应液浓缩至干直接用高效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得化合物5-22-B 13.83mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 10 90 28
2.00 10 90 28
18.00 90 10 28
结构表征数据如下：
5-22-B(6min LCMS出峰靠前，保留时间：2.612min)结构表征数据如下：
ESI-MS(m/z):520.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.49(d,J＝8.9Hz,1H),7.74(d,J＝11.9Hz,1H),7.31(s,1H),6.53(s,1H),5.60(s,1H),5.51(t,J＝5.9Hz,1H),5.43(s,2H),5.25–5.13(m,2H),3.97(d,J＝5.8Hz,2H),2.18(s,2H),2.04–1.91(m,4H),1.90–1.80(m,1H),1.23(s,6H),1.15–1.05(m,2H),0.87(t,J＝7.2Hz,4H),0.80–0.70(m,2H).
实施例二十(R)-3-(二甲胺)-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧代-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙酰胺和(S)-3-(二甲胺)-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧代-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙酰胺的合成
Figure PCTCN2022074328-appb-000172
步骤一：(9H-氟-9-基)甲基((S)-3-(((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)氨基)-2-羟基-3-氧丙基)氨基甲酸酯和(9H-氟-9-基)甲基((R)-3-(((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)氨基)-2-羟基-3-氧丙基)氨基甲酸酯的合成
25℃下，将(1S,9S)-1-氨基-9-乙基-5-氟-9-羟基-4-甲基-2,3,12,15-苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-10,13(1H,9H)-二酮甲磺酸盐(72mg,166μmol)和3-((((9H-芴-9-基)甲氧基)羰基)氨基)-2-羟基丙酸(65mg,199μmol,)溶于DMF(2mL)中，然后加入HATU(95mg,250μmol)和DIPEA(65mg,498μmol)，加完后，室温反应0.5hr；反应液浓缩至干直接用高效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得标题化合物(化合物1-10-1-A)24mg和(化合物1-10-1-B)28mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 15 85 28
2.00 15 85 28
18.00 90 10 28
化合物1-10-1-A(6min LCMS出峰靠前，保留时间：3.283min)结构表征数据如下：
ESI-MS(m/z):745.4[M+H] +.
化合物1-10-1-B(6min LCMS出峰靠后，保留时间：3.465min)结构表征数据如下：
ESI-MS(m/z):745.4[M+H] +.
步骤二：(R)-3-氨基-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧代-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙酰胺的合成和(S)-3-氨基-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧代-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙酰胺的合成
25℃下，将化合物1-10-1-B(28mg,37μmol)溶于DMF(2mL)中，然后加入二乙胺(1mL)，加完后，室温反应1.0hr；反应液浓缩至干得到产物粗品(化合物1-10-2-B)28mg，直接用于下一步反应。
25℃下，将化合物1-10-1-A(24mg,33μmol)溶于DMF(2mL)中，然后加入二乙胺(1mL)，加完后，室温反应1.0hr；反应液浓缩至干得到产物粗品(化合物1-10-2-A)24mg，直接用于下一步反应。
结构表征数据如下：
ESI-MS(m/z):523.2[M+H] +.
步骤三：(R)-3-(二甲胺)-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧代-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙酰胺的合成和(S)-3-(二甲胺)-N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲基-10,13-二氧代-1,2,3,9,10,12,13,15-八氢苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙酰胺的合成
25℃下，将化合物1-10-2-B(28mg,37μmol,70％)溶于甲醇中(2mL)中，然后加入甲醛水溶液(1mL)，加完后，室温反应16.0hr后，再加入氰基硼氢化钠(7.07mg,96.45μmol)，室温反应1.0hr；反应液浓缩至干直接用高效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得标题化合物(化合物1-10B)1.3mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：甲醇；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 15 85 28
2.00 15 85 28
18.00 90 10 28
化合物1-10-B(6min LCMS出峰靠前，保留时间：1.937min)结构表征数据如下：
ESI-MS(m/z):551.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.48(d,J＝8.6Hz,1H),8.31(s,2H),7.80(d,J＝11.0Hz,1H),7.31(s,1H),6.55(s,1H),5.54(s,1H),5.43(s,2H),5.34(d,J＝19.2Hz,1H),5.19(d,J＝19.1Hz,1H),4.09–4.06(m,1H),3.20–3.15(m,2H),2.59–2.53(m,1H),2.45–2.42(m,1H),2.42–2.38(s,3H),2.23–2.19(d,J＝7.0Hz,1H),2.13(s,6H),2.12–2.08(m,1H),2.02 –1.95(m,1H),1.90–1.85(m,2H),1.23(s,2H),0.88(d,J＝7.2Hz,3H).
25℃下，将化合物1-10-2-A(24mg,33μmol,70％)溶于甲醇中(2mL)中，然后加入甲醛水溶液(1mL)，加完后，室温反应16.0hr后，再加入氰基硼氢化钠(6.06mg,96.45μmol)，室温反应1.0hr；反应液浓缩至干直接用高效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得标题化合物(化合物1-10-A)4.44mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：甲醇；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 15 85 28
2.00 15 85 28
18.00 90 10 28
化合物1-10-A(6min LCMS出峰靠前，保留时间：1.920min)结构表征数据如下：
ESI-MS(m/z):551.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.58(d,J＝9.0Hz,1H),8.28(s,1H),7.84(d,J＝10.9Hz,1H),7.37(s,1H),6.61(s,1H),5.66–5.59(m,1H),5.49(s,2H),5.35(d,J＝19.1Hz,1H),5.17(d,J＝18.9Hz,1H),4.23–4.16(m,1H),3.23(d,J＝7.8Hz,2H),2.75–2.67(m,2H),2.45(s,3H),2.30(s,6H),2.27–2.17(m,2H),2.14–1.99(m,1H),1.98–1.87(m,2H),1.30(s,2H),0.93(t,J＝7.3Hz,3H).
实施例二十一(S)-14-(2-(环丙胺基)乙基)-7-乙基-7-羟基-7H-[1,3]二氧杂戊环并[4,5-g]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-8,11(10H,13H)-二酮(化合物4-14)
Figure PCTCN2022074328-appb-000173
步骤一：2-硝基-4,5-亚甲二氧基苯乙酮的合成
将化合物4-14-1(10.0g，60.92mmol)溶于硝基甲烷(100mL),搅拌下缓慢滴入浓硝酸(26mL)，室温反应2小时。TLC监控反应，原料少量残余，产物明显。缓慢滴入饱和碳酸氢钠水溶液中和反应，加入二氯甲烷萃取3次，有机相合并，饱和食盐水洗涤3次后干燥，浓缩得粗品。硅胶柱纯化(洗脱剂：0-20％乙酸乙酯/石油醚)得标题化合物9.8g。
步骤二：6-氨基-3,4-亚甲二氧基苯乙酮的合成
将化合物4-14-2(2.0g，9.56mmol)溶于乙酸乙酯(20mL)，加入10％钯碳(0.2g)，氢气置换并保护下搅拌反应4小时。过滤，滤液减压浓缩后得标题化合物粗品1.7g。
步骤三：6-乙酰氨基-3,4-亚甲二氧基苯乙酮的合成
将化合物4-14-3(1.7g,9.49mmol)溶于乙酸酐(17mL)，搅拌反应1小时。减压蒸除溶剂，加水搅拌，过滤，固体水洗后真空干燥得标题化合物粗品2.08g。
步骤四：(E)-N-(6-(3-(二甲氨基)丙烯酰)苯并[d][1,3]二氧杂戊环-5-基)乙酰胺的合成
将化合物4-14-4(1.88g，8.50mmol)溶于DMF-DMA(30mL)，升温120℃反应2小时。减压蒸除溶剂后得标题化合物粗品2.33g。
结构表征数据如下：
ESI-MS(m/z):277.2[M+1] +.
步骤五：(E)-N-(6-(3-(环丙基氨基)丙烯酰)苯并[d][1,3]二氧杂戊环-5-基)乙酰胺的合成
将化合物4-14-5(200mg，0.72mmol)溶于乙醇(5mL)，滴入环丙胺(413.3mg，7.24 mmol)，升温50℃反应16小时。减压蒸除溶剂得标题化合物粗品208mg。
结构表征数据如下：
ESI-MS(m/z):289.2[M+1] +.
步骤六：N-(6-(3-(环丙基氨基)丙酰基)苯并[d][1,3]二氧杂戊环-5-基)乙酰胺的合成
将化合物4-14-6(208mg，0.72mmol)溶于冰醋酸(4mL)，冰水浴冷却搅拌下加入硼氢化钠(13.65mg，0.36mmol)，恢复室温搅拌3小时。减压蒸除溶剂得标题化合物粗品209mg。
结构表征数据如下：
ESI-MS(m/z):291.1[M+1] +.
步骤七：(9H-芴-9-基)甲基(3-(6-乙酰氨基苯并[d][1,3]二氧杂戊环-5-基)-3-氧丙基)(环丙基)氨基甲酸酯的合成
将化合物4-14-7(200mg，0.69mmol)溶于1,4-二氧六环(20mL)和水(20mL)，搅拌下加入9-芴甲基-N-琥珀酰亚胺基碳酸酯(395mg，0.68mmol)和碳酸氢钠(231.5mg，2.76mmol)，室温反应2小时。加水和乙酸乙酯搅拌，静置分液，有机相用饱和食盐水洗涤后干燥浓缩，硅胶柱纯化(洗脱剂：30％乙酸乙酯/石油醚)，得标题化合物350mg。
步骤八：(9H-芴-9-基)甲基(3-(6-氨基苯并[d][1,3]二氧杂戊环-5-基)-3-氧丙基)(环丙基)氨基甲酸酯的合成
将化合物4-14-8(350mg，0.68mmol)溶于1,4-二氧六环(10mL)，滴入3N盐酸水溶液(10mL)，升温60℃搅拌反应16小时。加入水和乙酸乙酯搅拌，静置分液，有机相水洗后干燥浓缩，硅胶柱纯化(洗脱剂：33％乙酸乙酯/石油醚)，得标题化合物218mg。
步骤九：(S)-(9H-芴-9-基)甲基环丙基(2-(7-乙基-7-羟基-8,11-二氧基-8,10,11,13-四氢-7H-[1,3]二氧杂戊环并[4,5-g]吡喃并[3,4]:6,7]吲哚嗪并[1,2-b]喹啉-14-基)乙基)氨基甲酸酯的合成
将化合物4-14-9(40mg，0.085mmol)和(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃[3,4-f]吲哚嗪-3,6,10(4H)-三酮(24.62mg，0.094mmol)溶于甲苯(1mL)，加入对甲苯磺酸(2.93mg，0.017mmol)，升温120℃反应4小时。减压浓缩得标题化合物粗品59mg。
结构表征数据如下：
ESI-MS(m/z):698.1[M+1] +.
步骤十：(S)-14-(2-(环丙胺基)乙基)-7-乙基-7-羟基-7H-[1,3]二氧杂戊环并[4,5-g]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-8,11(10H,13H)-二酮的合成
将化合物4-14-10(59mg，0.085mmol)溶于DMF(1mL)，滴入二乙胺(0.5mL)，搅拌反应1小时。减压蒸除二乙胺，滴入3N盐酸酸化后，经高效液相制备纯化(纯化条件如下)，冻干得标题化合物的三氟乙酸盐12.66mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％三氟乙酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 20 80 28
2 20 80 28
18 80 20 28
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.73(s,2H),7.67(s,1H),7.57(s,1H),7.26(s,1H),6.54(s,1H),6.33(s,2H),5.44(s,2H),5.34(s,2H),3.40(s,4H),2.82(s,1H),1.91-1.81(m,2H),0.87(t,J＝7.2Hz,5H),0.79(d,J＝7.4Hz,2H).
ESI-MS(m/z):476.1[M+1] +.
实施例二十二(S)-7-乙基-7-羟基-14-(2-((2-甲氧基乙基)氨基)乙基)-7H-[1,3]二氧杂环并[4,5-g]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-8,11(10H,13H)-二酮(化合物4-15)
Figure PCTCN2022074328-appb-000174
步骤一：(E)-N-(6-(3-((2-甲氧基乙基)氨基)丙烯酰)苯并[d][1,3]二氧杂环-5-基)乙酰胺的合成
将化合物4-14-5(200mg，0.72mmol)溶于乙醇(5mL)，滴入2-甲氧基乙胺(543.7mg，7.24mmol)，升温50℃反应16小时。LCMS监控反应，原料有少量残余，产物明显。减压蒸除溶剂，得标题化合物221mg，直接进行下一步反应。
结构表征数据如下：
ESI-MS(m/z):307.1[M+1] +
步骤二：N-(6-(3-((2-甲氧基乙基)氨基)丙酰基)苯并[d][1,3]二氧杂环-5-基)乙酰胺的合成
将化合物4-15-1(200mg，0.65mmol)溶于冰醋酸(4mL)，冰水浴冷却搅拌下加入硼氢化钠(12.35mg，0.33mmol)，恢复室温搅拌3小时。LCMS监控反应，原料消失，产物明显。减压蒸除溶剂，得标题化合物200mg，直接进行下一步反应。
ESI-MS(m/z):309.1[M+1] +
步骤三：(9H-芴-9-基)甲基(3-(6-乙酰氨基苯并[d][1,3]二氧醇-5-基)-3-氧丙基)(2-甲氧基乙基)氨基甲酸酯的合成
将化合物4-15-2的粗品(200mg，0.65mmol)溶于1,4-二氧六环(20mL)和水(20mL)，搅拌下加入9-芴甲基-N-琥珀酰亚胺基碳酸酯(372mg，0.65mmol)和碳酸氢钠(231.5mg，2.76mmol)，室温反应2小时。TLC监控原料消失，产物明显。加水和乙酸乙酯搅拌，静置分液，有机相用饱和食盐水洗涤后干燥浓缩，硅胶柱纯化(洗脱剂：50％乙酸乙酯/石油醚)，得标题化合物180mg。
步骤四：((9H-芴-9-基)甲基(3-(6-氨基苯并[d][1,3]二恶英-5-基)-3-氧丙基)(2-甲氧基乙基)氨基甲酸酯的合成
将化合物4-15-3(180mg，0.68mmol)溶于1,4-二氧六环(5mL)，滴入3N盐酸水溶液(5mL)，升温60℃搅拌反应16小时。加入水和乙酸乙酯搅拌，静置分液，有机相水洗后干燥浓缩，硅胶柱纯化(洗脱剂：45％乙酸乙酯/石油醚)，得标题化合物132mg。
步骤五：(S)-(9H-芴-9-基)甲基(2-(7-乙基-7-羟基-8,11-二氧基-8,10,11,13-四氢-7H-[1,3]二氧杂环并[4,5-g]吡喃并[3'，4]：6,7]吲哚嗪并[1,2-b]喹啉-14-基)乙基)(2-甲氧基乙基)氨基甲酸酯的合成
将化合物4-15-4(130mg，0.266mmol)和rac-(4S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃并[3,4-f]吲哚嗪-3,6,10-三酮(70.05mg，0.266mmol)溶于甲苯(4mL)，加入对甲苯磺酸(9.16mg，0.053mmol)，升温120℃反应4小时。LCMS监控反应，原料消失，产物明显。减压浓缩，得标题化合物的粗品190mg。
结构表征数据如下：
ESI-MS(m/z):716.1[M+1] +
步骤六：(S)-7-乙基-7-羟基-14-(2-((2-甲氧基乙基)氨基)乙基)-7H-[1,3]二氧杂环并[4,5-g]吡喃并[3',4':6,7]吲哚嗪并[1,2-b]喹啉-8,11(10H,13H)-二酮的合成
将化合物4-15-5(190mg，0.265mmol)溶于DMF(3mL)，滴入二乙胺(2mL)，搅拌反应1小时。LCMS监控反应，原料消失，产物明显。减压蒸除二乙胺，滴入3N盐酸酸化后制备纯化，冻干得标题化合物99.28mg。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0 20 80 28
2 20 80 28
18 80 20 28
结构表征数据如下：
1H NMR(400MHz,DMSO-d 6):δ8.68(s,2H),7.68(s,1H),7.56(s,1H),7.26(s,1H),6.52(s,1H),6.32(s,2H),5.44(s,2H),5.31(s,2H),3.65–3.58(m,2H),3.42(d,J＝10.2Hz, 2H),3.36(s,3H),3.22(d,J＝4.0Hz,4H),1.94–1.80(m,2H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):494.2[M+1] +
实施例二十三 N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲氧基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基乙酰胺&N-((1R,9S)-9-乙基-5-氟-9-羟基-4-甲氧基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基乙酰胺
Figure PCTCN2022074328-appb-000175
步骤一：N-(3-溴-5-氟-4-甲氧基苯基)乙酰胺的合成
将3-溴-5-氟-4-甲氧基-苯胺(1.7g,7.73mmol)溶于四氢呋喃(30mL)中，然后加入三乙胺(2.35g,23.18mmol)和乙酸酐(1.18g,11.59mmol)，加毕，升温至50℃搅拌反应4小时；用高效液相质谱联用色谱检测反应；将反应液冷却至室温后，用乙酸乙酯(50mL)稀释，然后用清水(30mL)和饱和食盐水(30mL)各洗涤一次，分出有机相用无水硫酸钠干燥，过滤，滤液减压蒸干得粗品，经(石油醚：乙酸乙酯＝5:1)打浆纯化得标题化合物1.1g。
结构表征数据如下：
ESI-MS(m/z):262.0[M+H] +.
步骤二：(E)-4-(5-乙酰氨基-3-氟-2-甲氧基苯基)-3-丁烯酸的合成
将N-(3-溴-5-氟-4-甲氧基苯基)乙酰胺(1.1g,4.20mmol)和3-丁烯酸(397.47mg,4.62mmol)溶于1,4二氧六环(20mL)和水(5mL)的混合溶剂中，然后加入三乙胺(1.27g,12.59mmol)，三(邻甲基苯基)磷(127.75mg,419.73μmol)和醋酸钯(47.12mg,209.89μmol)，加毕，反应体系用氮气置换三次，并在氮气氛围下升温至100℃反应4小时；用高效液相 质谱联用色谱检测反应；反应液冷却至室温后，加入1mol/L的氢氧化钠水溶液(50mL)和乙酸乙酯(50mL)振荡分层。分出下层水相后，用4mol/L盐酸水溶液调节pH至3左右，然后用乙酸乙酯(40mL*2)萃取，合并有机相用饱和食盐水(40mL)洗涤，无水硫酸钠干燥，过滤，滤液减压蒸干得到标题化合物的粗品1.1g。
结构表征数据如下：
ESI-MS(m/z):268.1[M+H] +.
步骤三：4-(5-乙酰氨基-3-氟-2-甲氧基苯基)丁酸的合成
将(E)-4-(5-乙酰氨基-3-氟-2-甲氧基苯基)-3-丁烯酸的粗品(1.1g,4.12mmol,)溶于甲醇(20mL)，然后加入10％钯碳(100mg)，加毕，然后用氢气球置换反应体系三次，并在氢气氛围下反应4小时；用高效液相质谱联用色谱检测反应；反应液过滤，滤液减压浓缩干得标题化合物的粗品1.05g。
结构表征数据如下：
ESI-MS(m/z):270.1[M+H] +.
步骤四：N-(3-氟-4-甲氧基-8-氧基-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将4-(5-乙酰氨基-3-氟-2-甲氧基苯基)丁酸的粗品(1.1g,4.09mmol)溶于三氟乙酸(10mL)中，降温至5℃后，缓慢加入三氟乙酸酐(4.29g,20.43mmol)，加毕，自然升至室温反应2小时；用高效液相质谱联用色谱检测反应；反应液缓慢倒入水(60mL)中,然后用乙酸乙酯(40mL*3)萃取，合并有机相，用饱和碳酸氢钠水溶液洗涤至中性，然后再用饱和食盐水(40mL)洗涤，无水硫酸钠干燥有机相，然后过滤，滤液减压蒸干得粗品，经快速硅胶柱(乙酸乙酯:石油醚＝0-40％)纯化得标题化合物503mg。
结构表征数据如下：
ESI-MS(m/z):252.1[M+H] +.
步骤五：N-(3-氟-7-(羟基亚氨基)-4-甲氧基-8-氧基-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将四氢呋喃(15mL)和叔丁醇(4mL)加入反应液瓶中，冰浴降温至5℃后，加入叔丁醇钾(491.26mg,4.38mmol)，然后将N-(3-氟-4-甲氧基-8-氧基-5,6,7,8-四氢萘-1-基)乙酰胺(500mg,1.99mmol)溶于四氢呋喃(5mL)中，并缓慢滴加入反应液，10分钟后再加入亚硝酸异戊酯(373.01mg,3.18mmol)，加毕，保持5℃反应1小时；用高效液相质谱联用色谱检测反应；反应液用饱和氯化铵水溶液(50mL)淬灭后，用乙酸乙酯(40*2)萃取，合并有机相，用饱和食盐水(40mL)洗涤，无水硫酸钠干燥有机相，然后过滤，滤液减压蒸干得标题化合物的粗品550mg。
结构表征数据如下：
ESI-MS(m/z):281.1[M+H] +.
步骤六：N-(7-氨基-3-氟-4-甲氧基-8-氧基-5,6,7,8-四氢萘-1-基)乙酰胺的合成
将N-(3-氟-7-(羟基亚氨基)-4-甲氧基-8-氧基-5,6,7,8-四氢萘-1-基)乙酰胺的粗品(520mg,1.86mmol)溶于甲醇(10mL)和四氢呋喃(10mL)的混合溶液中，然后加入1mol/L的 盐酸水溶液(3.71mL)和10％钯碳(50mg)，加毕，用氢气球置换反应体系三次，并在氢气氛围下室温反应1小时；用高效液相质谱联用色谱检测反应；反应液过滤，滤液减压浓缩干得标题化合物的盐酸盐粗品551mg。
结构表征数据如下：
ESI-MS(m/z):267.1[M+H] +.
步骤七：(9H-芴-9-基)甲基(8-乙酰胺-6-氟-5-甲氧基-1-氧基-1,2,3,4-四氢萘-2-基)氨基甲酸酯的合成
将N-(7-氨基-3-氟-4-甲氧基-8-氧基-5,6,7,8-四氢萘-1-基)乙酰胺的盐酸盐粗品(550mg,1.64mmol)溶于1,4-二氧六环(15mL)中，然后加入碳酸氢钠(549.45mg,6.54mmol)、水(5mL)和9-芴甲基-N-琥珀酰亚胺基碳酸酯(1.12g,1.96mmol)，加毕，室温下搅拌反应2小时；用高效液相质谱联用色谱检测反应；反应液倒入水(50mL)中，然后用乙酸乙酯(40mL*2)萃取，合并有机相，用饱和食盐水(40mL)洗涤，无水硫酸钠干燥有机相，过滤，滤液减压整干得粗品。粗品经C18(乙腈/0.05％甲酸水溶液,20％乙腈～100％乙腈)反相柱纯化得标题化合物410mg。
结构表征数据如下：
ESI-MS(m/z):489.1[M+H] +.
步骤八：(9H-芴-9-基)甲基(8-氨基-6-氟-5-甲氧基-1-氧基-1,2,3,4-四氢萘-2-基)氨基甲酸酯的合成
将(9H-芴-9-基)甲基(8-乙酰胺-6-氟-5-甲氧基-1-氧基-1,2,3,4-四氢萘-2-基)氨基甲酸酯(410mg,839.29μmol)溶于二氧六环(10mL)中，加入12mol/L的浓盐酸(2mL)，加毕，升温至70℃反应2小时；用高效液相质谱联用色谱检测反应；反应液倒入水(30mL)中，然后用乙酸乙酯(30mL*2)萃取，合并有机相，用饱和食盐水(30mL)洗涤，无水硫酸钠干燥有机相，过滤，滤液减压整干得粗品。经快速硅胶(乙酸乙酯:石油醚＝0-60％)纯化得标题化合物351mg。
结构表征数据如下：
ESI-MS(m/z):447.1[M+H] +.
步骤九：(9H-芴-9-基)甲基((9S)-9-乙基-5-氟-9-羟基-4-甲氧基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)氨基甲酸酯的合成
将(S)-4-乙基-4-羟基-7,8-二氢-1H-吡喃并[3,4-f]吲哚嗪-3,6,10(4H)-三酮(247.64mg,940.71μmol)和(9H-芴-9-基)甲基(8-氨基-6-氟-5-甲氧基-1-氧基-1,2,3,4-四氢萘-2-基)氨基甲酸酯(350mg,783.93μmol)加入甲苯(15mL)中，然后再加人对甲苯磺酸(134.84mg,783.93μmol)，加毕，升温至135℃反应4小时，反应液直接135℃下减压蒸干得粗品；粗品经快速硅胶(甲醇:二氯甲烷＝0-6％)柱纯化得标题化合物358mg。
结构表征数据如下：
ESI-MS(m/z):674.2[M+H] +.
步骤十：(9S)-1-氨基-9-乙基-5-氟-9-羟基-4-甲氧基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4'：6,7]吲哚嗪[1,2-b]喹啉-10,13-二酮的合成
将(9H-芴-9-基)甲基((9S)-9-乙基-5-氟-9-羟基-4-甲氧基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)氨基甲酸酯(358mg,531.41μmol)溶于N,N-二甲基甲酰胺(4mL)中，然后加入二乙胺(0.4mL)，加毕，室温反应0.5小时；用高效液相质谱联用色谱检测反应；反应液减压蒸干得粗品，粗品经乙酸乙酯打浆纯化得标题化合物220mg。
结构表征数据如下：
ESI-MS(m/z):452.1[M+H] +.
步骤十一：N-((1S,9S)-9-乙基-5-氟-9-羟基-4-甲氧基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基乙酰胺&N-((1R,9S)-9-乙基-5-氟-9-羟基-4-甲氧基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基乙酰胺的合成
将(9S)-1-氨基-9-乙基-5-氟-9-羟基-4-甲氧基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4'：6,7]吲哚嗪[1,2-b]喹啉-10,13-二酮(50mg,110.76μmol)和2-羟基乙酸(16.85mg,221.51μmol)溶于N,N-二甲基甲酰胺(2mL)中，然后加人HATU(84.17mg,221.51μmol)和N,N-二异丙基乙胺(42.94mg,332.27μmol)，加毕，室温反应0.5小时；用高效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得单一构型标题化合物的两个异构体(5-34-A：6.22mg，5-34-B：9.81mg)
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 10 90 28
2.00 10 90 28
18.00 90 10 28
5-34-A结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.53(d,J＝9.0Hz,1H),7.89(d,J＝12.3Hz,1H),7.33(s,1H),5.58(q,J＝7.5,7.0Hz,1H),5.42(d,J＝2.1Hz,2H),5.25–5.09(m,2H),3.98(s,2H),3.96(d,J＝1.1Hz,3H),3.30–3.10(m,2H),2.15(q,J＝7.4Hz,2H),1.93–1.80(m,2H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):510.2[M+H] +.
5-34-B结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.55(d,J＝9.0Hz,1H),7.90(d,J＝12.3Hz,1H),7.34(s,1H),5.59(q,J＝7.4,6.8Hz,1H),5.43(s,2H),5.25–5.11(m,2H),3.99(s,2H),3.96 (d,J＝1.1Hz,3H),3.30–3.13(m,2H),2.15(q,J＝6.4Hz,2H),1.93–1.82(m,2H),0.88(t,J＝7.3Hz,3H).
ESI-MS(m/z):510.2[M+H] +.
实施例二十四(2R)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙胺&(2R)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙胺
Figure PCTCN2022074328-appb-000176
步骤一：(2R)-N-((1S,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙胺&(2R)-N-((1R,9S)-5-氯-9-乙基-9-羟基-4-甲基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4:6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙胺的合成
将(9S)-1-氨基-5-氯-9-乙基-9-羟基-4-甲基-1,2,3,9,12,15-六氢-10H,13H-苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-10,13-二酮(50mg,110.64μmol)和(2R)-2-羟基丙酸(19.93mg,221.29μmol)溶于N,N-二甲基甲酰胺(2mL)中，然后加人HATU(84.09mg,221.29μmol)和N,N-二异丙基乙胺(42.90mg,331.93μmol)，加毕，室温反应0.5小时；用高效液相质谱联用色谱检测反应；反应液直接用制备高效液相色谱纯化得标题化合物的两个异构体(2-27-A：5.73mg，2-27-B：7.59mg)
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％三氟乙酸)
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 30 90 28
2.00 30 90 28
18.00 90 10 28
2-27-A结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.45(d,J＝9.1Hz,1H),8.14(s,1H),7.30(s,1H),6.54(s,1H),5.63(s,1H),5.56(q,J＝8.0Hz,1H),5.42(s,2H),5.25(d,J＝19.0Hz,1H),5.08(d,J＝19.0Hz,1H),4.13(q,J＝6.7Hz,1H),3.27–3.12(m,2H),2.51(s,3H),2.23–2.13(m,2H),1.92–1.80(m,2H),1.41(d,J＝6.8Hz,3H),0.87(t,J＝7.3Hz,3H).
ESI-MS(m/z):524.2[M+H] +.
2-27-B结构表征数据如下：
1H NMR(400MHz,DMSO-d 6)δ8.40(d,J＝8.9Hz,1H),8.15(s,1H),7.31(s,1H),6.54(s,1H),5.55–5.49(m,1H),5.43(d,J＝2.2Hz,2H),5.18(q,J＝19.0Hz,2H),4.13(q,J＝6.6Hz,1H),3.24–3.12(m,2H),2.51(s,3H),2.22–2.10(m,2H),1.92–1.82(m,2H),1.30(d,J＝6.7Hz,3H),0.88(t,J＝7.3Hz,3H).
ESI-MS(m/z):524.2[M+H] +.
实施例二十五：(R)-N-((1S,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙胺和(R)-N-((1R,9S)-4-氯-9-乙基-5-氟-9-羟基-10,13-二氧基-2,3,9,10,13,15-六氢-1H,12H-苯并[de]吡喃并[3',4':6,7]吲哚嗪[1,2-b]喹啉-1-基)-2-羟基丙胺的制备
Figure PCTCN2022074328-appb-000177
25℃下，将3-1-A(20.0mg,43.9μmol)和D-乳酸(7.90mg,87.8μmol)溶于DMF(1.0mL)中，然后加入HATU(33.4mg,87.8μmol)和DIPEA(17.0mg,131.6μmol)，室温反应2hr；将反应液浓缩掉大部分DMF，残留用制备高效液相色谱纯化得到化合物3-26-A(15.4mg，收率64％)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
保留时间：5.3～6.2min
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 30 70 28
2.00 30 70 28
18.00 90 10 28
结构表征数据如下：
MS m/z(ESI):528.2[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.52(d,J＝9.2Hz,1H),8.05(d,J＝10.0Hz,1H),7.33(s,1H),6.55(s,1H),5.62–5.59(m,2H),5.43(s,2H),5.28–5.10(m,2H),4.13–4.11(m,1H),3.41–3.38(m,1H),3.28–3.22(m,1H),2.20–2.18(m,2H),1.92–1.80(m,2H),1.40(d,J＝6.8Hz,3H),0.87(t,J＝7.2Hz,3H).
25℃下，将3-1-B(20.0mg,43.9μmol)和D-乳酸(7.90mg,87.8μmol)溶于DMF(1.0mL)中，然后加入HATU(33.4mg,87.8μmol)和DIPEA(17.0mg,131.6μmol)，室温反应2hr；将反应液浓缩掉大部分DMF，残留用制备高效液相色谱纯化得到化合物3-26-B(4.8mg，收率20％)。
色谱柱：SunFire Prep C18 OBD 19mm×150mm×5.0μm
流动相A：乙腈；流动相B：水(0.05％甲酸)
保留时间：7.5～8.5min
时间[min] 流动相A[％] 流动相B[％] 流速[mL/min]
0.00 30 70 28
4.00 30 70 28
20.00 90 10 28
结构表征数据如下：
MS m/z(ESI):528.2[M+H] +
1H NMR(400MHz,DMSO-d 6)δ8.48(d,J＝8.8Hz,1H),8.06(d,J＝10.4Hz,1H),7.34(s,1H),6.57(br,1H),5.61–5.55(m,1H),5.48–5.39(m,2H),5.27–5.16(m,2H),4.15–4.10(m,1H),3.32–3.21(m,3H),2.23–2.16(m,2H),1.92–1.81(m,2H),1.30(d,J＝6.4Hz,3H),0.87(t,J＝7.2Hz,3H).
生物学评价
一、肿瘤细胞增殖抑制试验
1.化合物对HT29细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞HT29，用胰酶消化细胞，离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表1。
表1.肿瘤细胞来源
细胞名称 肿瘤类型 来源
HT29 人结肠癌细胞 中国科学院细胞库
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50yL，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting- Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表2所示。
(2)数据结果
表2.HT29细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 11.27
实施例一(1-1-A) 1.28
实施例一(1-1-B) 1.54
实施例三(2-1-A) 5.03
实施例三(2-2-B) 7.52
实施例四(2-7-A) 1.56
实施例四(2-7-B) 5.44
实施例五(2-12-A) 3.01
实施例五(2-12-B) 2.34
实施例五(2-12-C) 1.49
实施例五(2-12-D) 3.29
实施例六(2-17-A) 1.24
实施例六(2-17-B) 1.59
实施例七(2-20-A) 1.29
实施例七(2-20-B) 4.03
实施例八(3-1-A) 0.16
实施例八(3-1-B) 0.71
实施例九(3-4-A) 5.55
实施例九(3-4-B) 4.98
实施例十(4-12) 4.79
实施例十一(4-10) 0.52
实施例十二(5-13-A) 0.88
实施例十三(5-7-A) 3.13
实施例十三(5-7-B) 4.66
实施例十四(5-16-A) 3.87
实施例二十(1-10-A) 61.60
实施例二十(1-10-B) 10.26
实施例二十一(4-14) 0.56
实施例二十二(4-15) 5.20
测试结果表明，表2中本发明的化合物对HT29结肠癌细胞增殖具有较强的增殖抑制作用。
2.化合物对A549细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞A549，用胰酶消化细胞，离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表3。
表3.肿瘤细胞来源
细胞名称 肿瘤类型 来源
A549 人肺癌细胞 中国科学院细胞库
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50 0L，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表4所示。
(2)数据结果
表4.A549细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 94.18
实施例一(1-1-A) 66.56
实施例四(2-7-A) 6.97
实施例四(2-7-B) 16.39
实施例五(2-12-A) 6.96
实施例五(2-12-B) 5.52
实施例五(2-12-C) 6.64
实施例七(2-20-A) 8.14
实施例七(2-20-B) 23.06
实施例十一(4-10) 68.48
测试结果表明，表4中本发明的化合物对A549人肺癌细胞增殖具有显著的抑制作用。
3.化合物对NCI-H1806细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞HCC1806，用胰酶消化细胞， 离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表5。
表5.肿瘤细胞来源
细胞名称 肿瘤类型 来源
HCC1806 人乳腺鳞状癌细胞 ATCC
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50yL，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表6所示。
(2)数据结果
表6.HCC1806细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 3.03
实施例三(2-1-A) 1.43
实施例四(2-7-A) 1.25
实施例五(2-12-A) 1.40
实施例五(2-12-B) 1.07
实施例五(2-12-C) 2.08
实施例十二(5-13-A) 0.96
实施例十六(3-12-A) 0.16
实施例十六(3-12-B) 1.76
实施例十六(3-12-C) 1.91
实施例十六(3-12-D) 1.53
实施例十七(3-7-B) 1.94
实施例十八(3-17-A) 1.70
实施例十八(3-17-B) 2.26
实施例十九(5-22-A) 2.57
实施例十九(5-22-B) 3.71
实施例二十四(2-27-A) 4.55
实施例二十四(2-27-B) 3.93
实施例二十五(3-26-A) 1.27
实施例二十五(3-26-B) 4.93
测试结果表明，表6中本发明的化合物对HCC1806人乳腺鳞状癌细胞增殖具有显著的抑制作用。
4.化合物对SKOV-3细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞SKOV-3，用胰酶消化细胞，离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表7。
表7.肿瘤细胞来源
细胞名称 肿瘤类型 来源
SKOV-3 人卵巢癌细胞 南京科佰生物
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50μ0，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表8所示。
(2)数据结果
表8.SKOV-3细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 17.14
实施例一(1-1-A) 7.81
实施例三(2-1-A) 7.12
实施例四(2-7-A) 8.64
实施例四(2-7-B) 9.47
实施例五(2-12-A) 6.85
实施例五(2-12-B) 4.99
实施例五(2-12-C) 6.52
实施例七(2-20-A) 6.79
实施例十二(5-13-A) 2.09
实施例二十一(4-14) 2.16
测试结果表明，表8中本发明的化合物对SKOV-3人卵巢癌细胞增殖具有显著的抑制作用。
5.化合物对NCI-H358细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞NCI-H358，用胰酶消化细胞，离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表9。
表9.肿瘤细胞来源
细胞名称 肿瘤类型 来源
NCI-H358 人非小细胞肺癌细胞 南京科佰生物
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50μ0，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表10所示。
(2)数据结果
表10.NCI-H358细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 58.84
实施例一(1-1-A) 65.22
实施例二(1-7-A) 11.38
实施例二(1-7-B) 51.52
实施例三(2-1-A) 68.62
实施例四(2-7-A) 35.42
实施例四(2-7-B) 51.06
实施例五(2-12-A) 17.65
实施例五(2-12-B) 15.76
实施例五(2-12-C) 23.08
实施例七(2-20-A) 18.56
实施例十二(5-13-A) 7.01
实施例十六(3-12-A) 15.45
实施例十六(3-12-B) 19.77
实施例十六(3-12-C) 28.29
实施例十六(3-12-D) 20.75
实施例十七(3-7-A) 49.38
实施例十七(3-7-B) 22.22
实施例十八(3-17-A) 13.34
实施例十八(3-17-B) 37.34
实施例十九(5-22-A) 29.94
实施例十九(5-22-B) 61.36
实施例二十一(4-14) 11.98
实施例二十二(4-15) 17.76
实施例二十四(2-27-A) 56.46
实施例二十四(2-27-B) 69.12
实施例二十五(3-26-A) 31.64
测试结果表明，表10中本发明的化合物对NCI-H358人非小细胞肺癌细胞增殖具有显著的抑制作用。
6.化合物对NCI-N87细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞NCI-N87，用胰酶消化细胞，离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表11。
表11.肿瘤细胞来源
细胞名称 肿瘤类型 来源
NCI-N87 人胃癌细胞 ATCC
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50μL，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表12所示。
(2)数据结果
表12.NCI-N87细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 7.18
实施例一(1-1-A) 4.58
实施例二(1-7-A) 2.70
实施例二(1-7-B) 5.48
实施例四(2-7-A) 3.76
实施例五(2-12-A) 4.91
实施例五(2-12-B) 4.97
实施例五(2-12-C) 4.51
实施例七(2-20-A) 4.54
实施例八(3-1-A) 0.37
实施例八(3-1-B) 1.34
实施例九(3-4-A) 4.38
实施例九(3-4-B) 6.51
实施例十一(4-10) 2.29
实施例十二(5-13-A) 2.33
实施例十三(5-7-A) 6.03
实施例十三(5-7-B) 7.31
实施例十六(3-12-A) 0.44
实施例十六(3-12-B) 3.45
实施例十六(3-12-C) 3.99
实施例十六(3-12-D) 3.03
实施例十七(3-7-A) 5.32
实施例十七(3-7-B) 4.54
实施例十八(3-17-A) 3.01
实施例十八(3-17-B) 4.19
实施例十九(5-22-A) 3.35
实施例十九(5-22-B) 5.51
实施例二十四(2-27-A) 8.32
实施例二十四(2-27-B) 14.67
实施例二十五(3-26-A) 1.23
实施例二十五(3-26-B) 25.34
测试结果表明，表12中本发明的化合物对NCI-N87人胃癌细胞增殖具有显著的抑制作用。
7.化合物对Hela细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞Hela，用胰酶消化细胞，离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表13。
表13.肿瘤细胞来源
细胞名称 肿瘤类型 来源
Hela 人宫颈癌细胞 南京科佰生物
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的 生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50μL，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表14所示。
(2)数据结果
表14.Hela细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 17.57
实施例一(1-1-A) 4.57
实施例一(1-1-B) 3.65
实施例三(2-1-A) 16.98
实施例四(2-7-A) 13.77
实施例五(2-12-A) 13.25
实施例五(2-12-B) 16.68
实施例七(2-20-A) 11.17
实施例十一(4-10) 3.04
测试结果表明，表14中本发明的化合物对Hela人宫颈癌细胞增殖具有显著的抑制作用。
8.化合物对HCC70细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞HCC70，用胰酶消化细胞，离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表15。
表15.肿瘤细胞来源
细胞名称 肿瘤类型 来源
HCC70 人乳腺癌细胞 南京科佰生物
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50μ0，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按 照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表16所示。
(2)数据结果
表16.HCC70细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 226.10
实施例一(1-1-A) 224.85
实施例四(2-7-A) 150.70
实施例四(2-7-B) 192.20
实施例五(2-12-A) 170.93
实施例五(2-12-B) 176.16
实施例五(2-12-C) 210.68
实施例七(2-20-A) 104.99
实施例十二(5-13-A) 36.65
测试结果表明，表16中本发明的化合物对HCC70人乳腺癌细胞增殖具有显著的抑制作用。
9.化合物对MDA-MB-231细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞MDA-MB-231，用胰酶消化细胞，离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表17。
表17.肿瘤细胞来源
细胞名称 肿瘤类型 来源
MDA-MB-231 人乳腺癌细胞 南京科佰生物
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50)m，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表18所示。
(2)数据结果
表18.MDA-MB-231细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 381.70
实施例一(1-1-A) 363.60
实施例三(2-1-A) 299.70
实施例四(2-7-A) 123.50
实施例五(2-12-A) 123.50
实施例九(3-4-A) 284.80
实施例十二(5-13-A) 332.60
实施例十三(5-7-B) 172.40
实施例二十(1-10-A) 97.57
实施例二十(1-10-B) 27.24
实施例二十二(4-15) 175.40
测试结果表明，表18中本发明的化合物对MDA-MB-231人乳腺癌细胞增殖具有显著的抑制作用。
10.化合物对Jeko-1细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞Jeko-1，将细胞离心后重悬并细胞计数，最后调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表19。
表19.肿瘤细胞来源
细胞名称 肿瘤类型 来源
Jeko-1 人套细胞淋巴瘤细胞 ATCC
本发明化合物和肿瘤细胞共孵育：待细胞铺板后，按照一定的稀释比例将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50 0赞，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC50)，检测结果如表20所示。
(2)数据结果
表20.Jeko-1细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 1.25
实施例一(1-1-A) 1.33
实施例二(1-7-A) 0.15
实施例二(1-7-B) 0.66
实施例四(2-7-A) 0.28
实施例九(3-4-A) 0.27
实施例九(3-4-B) 0.15
实施例十一(4-10) 0.45
实施例十三(5-7-A) 0.16
实施例十三(5-7-B) 0.18
实施例二十一(4-14) 0.34
实施例二十二(4-15) 0.11
测试结果表明，表20中本发明的化合物对Jeko-1人套细胞淋巴瘤细胞增殖具有显著的抑制作用。
11.化合物对MDA-MB-453细胞增殖的抑制作用
(1)细胞铺板：首先采用相应的培养基培养肿瘤细胞MDA-MB-453，用胰酶消化细胞，离心后重悬细胞计数，调整细胞至合适的浓度进行铺板。肿瘤细胞来源见表21。
表21.肿瘤细胞来源
细胞名称 肿瘤类型 来源
MDA-MB-453 人乳腺癌细胞 康诺泰
本发明化合物和肿瘤细胞共孵育：待细胞贴壁后，移除细胞中培养基，将稀释好的生物活性分子(本发明化合物)加入到上述板孔中，孵育72h。
体外细胞活性检测：孵育结束后，每孔加入Cell Counting-Lite TM 2.0试剂(Vazyme/诺唯赞)50 0赞，避光振荡混匀，反应10min后即可进行检测，酶标仪(厂家：BMG，型号：PHERAStar-FS)读数。利用不含细胞的培养基(含Cell Counting-Lite TM)获得背景RLU，含有细胞的培养基(含Cell Counting-Lite TM)获得溶媒RLU。细胞抑制率＝1-(样品RLU-样品RLU)/(溶媒RLU-背景RLU)×100％，按照四参数模型拟合曲线，计算化合物的半数抑制浓度(IC 50)，检测结果如表22所示。
(2)数据结果
表22.MDA-MB-453细胞增殖抑制活性
名称 IC 50(nM)
对比化合物一 6.99
实施例十六(3-12-A) 0.47
实施例十六(3-12-B) 4.32
实施例十六(3-12-C) 5.52
实施例十六(3-12-D) 3.98
实施例十七(3-7-A) 6.23
实施例十七(3-7-B) 4.87
实施例十八(3-17-A) 3.32
实施例十八(3-17-B) 6.16
实施例十九(5-22-A) 7.34
实施例十九(5-22-B) 5.67
实施例二十四(2-27-A) 11.83
实施例二十五(3-26-A) 3.95
测试结果表明，表22中本发明的化合物对MDA-MB-453人乳腺癌细胞增殖具有显著的抑制作用。
对比化合物一和对比化合物二的结构如下所示：
Figure PCTCN2022074328-appb-000178
二、抗体偶联试验
ADC D-L-15样品的偶联制备如下：
取1.036mL hIgG抗体(抗鸡溶菌酶抗体，19.3mg/mL)，用0.1M依地酸二钠的溶液(pH 7.6)稀释，然后用1M Na 2HPO 4溶液调pH至7.6，加入2.4倍物质的量的10mM TCEP(三(2-羧乙基)膦)溶液(pH 7.6)混匀，室温放置90min。向上述溶液体系加入5倍物质的量的溶解在二甲基亚砜的化合物D-L-15，混匀，室温静置2h，完毕后采用NAP-5凝胶柱(Cytiva)将缓冲液置换为pH 6.0的10mM组氨酸缓冲溶液，然后添加蔗糖和吐温20，混匀，得到抗体药物缀合物ADC D-L-15(1.77mL，8.60mg/mL)。
Figure PCTCN2022074328-appb-000179
利用LC-MS测定ADC D-L-15分子量，计算药物/抗体比DAR值为4.11，如表23、表24所示。
表23：ADC D-L-15实测分子量
Figure PCTCN2022074328-appb-000180
表24：ADC D-L-15的DAR值
Figure PCTCN2022074328-appb-000181
色谱测定条件：
液相色谱柱：Thermo MAbPac RP 3.0*100mm；
流动相A：0.1％FA/H 2O；流动相B：0.1％FA/ACN；
流速：0.25mL/min；样品室温度：8℃；柱温：60℃；进样量：2μL；
时间(分钟) 2 20 22 25 26 30
流动相A(体积％) 75 60 5 5 75 75
流动相B(体积％) 25 40 95 95 25 25
质谱测定条件：
质谱型号：AB Sciex Triple TOF 5600+；
GS1 35；GS2 35；CUR 30；TEM 350；ISVF 5500；DP 250；CE 10；Accumulation time 0.5s；m/z 600-4000；Time bins to sum 40。
ADC 3-4-04-A样品的偶联制备如下：
取0.518mL hIgG抗体(抗鸡溶菌酶抗体，19.3mg/mL)，用0.1M依地酸二钠的溶液(pH 7.6)稀释，然后用1M Na 2HPO 4溶液调pH至7.6，加入5.5倍物质的量的10mM TCEP(三(2-羧乙基)膦)溶液(pH 7.6)混匀，室温放置90min。向上述溶液体系加入10倍物质的量的溶解在二甲基亚砜的化合物3-4-04-A，混匀，室温静置2h，完毕后采用NAP-5凝胶柱(Cytiva)将缓冲液置换为pH 6.0的10mM组氨酸缓冲溶液，然后添加蔗糖和吐温20，混匀，得到抗体药物缀合物ADC 3-4-04-A(1.50mL，5.60mg/mL)。
Figure PCTCN2022074328-appb-000182
利用LC-MS测定ADC 3-4-04-A分子量，计算药物/抗体比DAR值为7.47，如表25、表26所示，色谱测定条件同ADC D-L-15。
表25：ADC 3-4-04-A实测分子量
Figure PCTCN2022074328-appb-000183
表26：ADC 3-4-04-A的DAR值
Figure PCTCN2022074328-appb-000184
ADC 3-4-04-B样品的偶联制备如下：
取0.518mL hIgG抗体(抗鸡溶菌酶抗体，19.3mg/mL)，用0.1M依地酸二钠的溶液(pH 7.6)稀释，然后用1M Na 2HPO 4溶液调pH至7.6，加入5.5倍物质的量的10mM TCEP(三(2-羧乙基)膦)溶液(pH 7.6)混匀，室温放置90min。向上述溶液体系加入10倍物质的量的溶解在二甲基亚砜的化合物3-4-04-B，混匀，室温静置2h，完毕后采用NAP-5凝胶柱(Cytiva)将缓冲液置换为pH 6.0的10mM组氨酸缓冲溶液，然后添加蔗糖和吐温20，混匀，得到抗体药物缀合物ADC 3-4-04-B(1.50mL，5.60mg/mL)。
Figure PCTCN2022074328-appb-000185
利用LC-MS测定ADC 3-4-04-B分子量，计算药物/抗体比DAR值为8.04，如表27、表28所示，色谱测定条件同ADC D-L-15。
表27：ADC 3-4-04-B实测分子量
Figure PCTCN2022074328-appb-000186
表28：ADC 3-4-04-B的DAR值
Figure PCTCN2022074328-appb-000187
上述实验证明：本发明的细胞毒性药物-连接子类化合物可以顺利与抗体偶联得到抗体药物缀合物。
尽管本发明的具体实施方式已经得到详细的描述，本领域技术人员将会理解，根据已经公开的所有教导，可以对那些细节进行各种修改和替换，这些改变均在本发明的保护范围之内。本发明的全部范围由所附权利要求及其任何等同物给出。

Claims (14)

  1. 化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，其中所述化合物具有如下所示结构：
    Figure PCTCN2022074328-appb-100001
    其中，
    R 1和R 2各自独立地选自氢、卤素、C 1-6烷基、C 1-6烷氧基、C 1-6卤代烷基、羟基、氰基和C 3-6环烷基；或者，R 1和R 2与相邻碳原子连接成5-6元含氧杂环；
    R 3为氢或者与R 1邻位碳原子相连成六元碳环；
    A选自
    Figure PCTCN2022074328-appb-100002
    中的一种；
    R 4选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3-6环烷基和3-6元杂环基；
    R 5和R 6各自独立地选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷胺基烷基、C 1-6烷氧基烷基、C 2-6烯基、C 2-6炔基、C 3-6环烷基、3-6元杂环基、芳基和杂芳基；或者R 5和R 6与相邻碳原子连接成3-6元碳环或杂环；
    m＝1或2。
  2. 权利要求1所述的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，其中所述化合物具有式(I)的结构：
    Figure PCTCN2022074328-appb-100003
    上式(I)中，R x选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3-6环烷基和3-6元杂环基；
    R y和R z不同时为氢，且独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 2-6烯基、C 2-6炔基、C 1-6烷胺基烷基、C 1-6烷氧基烷基、3-6元杂环基烷基和3-6元杂环基；
    优选地，R x选自氢和C 1-6烷基；优选地，R x为氢；
    优选地，R y和R z不同时为氢，且独立选自氢、
    Figure PCTCN2022074328-appb-100004
    二甲胺基亚甲基、吗啉基亚甲基和甲氧基亚甲基；
    优选地，R y为氢，R z选自
    Figure PCTCN2022074328-appb-100005
    二甲胺基亚甲基、吗啉基亚甲基和甲氧基亚甲基。
  3. 权利要求1所述的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，其中所述化合物具有式(II)的结构：
    Figure PCTCN2022074328-appb-100006
    上式(II)中，A’选自
    Figure PCTCN2022074328-appb-100007
    中的一种；
    R x’选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3-6环烷基和3-6元杂环基；
    R y’和R z’独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 1-6烷氧烷基、C 3-6环烷基、3-6元杂环基、3-6元杂环基烷基、C 2-6烯基、C 2-6炔基、芳基和杂芳基，或者R y’和R z’与相邻碳原子连接成3-6元环；
    优选地，R x’选自氢和C 1-6烷基；
    优选地，R y’和R z’独立选自氢、C 1-6烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基和C 2-6烯基，或者R y’和R z’与相邻碳原子连接成3-6元环烷基；
    优选地，R y’选自氢和C 1-6烷基，R z’选自氢、C 1-6烷基和C 3-6环烷基，或者R y’和R z’与相邻碳原子连接成3-6元环烷基。
  4. 权利要求1所述的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，其中所述化合物具有式(III)的结构：
    Figure PCTCN2022074328-appb-100008
    上式(III)中，A”选自
    Figure PCTCN2022074328-appb-100009
    中的一种；
    R x”选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3-6环烷基和3-6元杂环基；
    R y”和R z”独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 1-6烷氧烷基、C 3-6环烷基、3-6元杂环基、3-6元杂环基烷基、4-6元杂环基、C 2- 6烯基、C 2-6炔基、芳基和杂芳基，或者R y”和R z”与相邻碳原子连接成3-6元环；
    优选地，R x”为氢；
    优选地，R y”和R z”各自独立地选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基和乙烯基，或者R y”和R z”与相邻碳原子连接成3-6元环；
    优选地，R y”为氢，R z”选自氢、C 1-6烷基、C 3-6环烷基和乙烯基，或者R y”和R z”与相邻碳原子连接成3-6元碳环。
  5. 权利要求1所述的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，其中所述化合物具有式(IV)的结构：
    Figure PCTCN2022074328-appb-100010
    上式(IV)中，
    R a和R b独立选自氢、卤素、C 1-6烷基、C 1-6羟烷基、C 1-6烷氧烷基、C 1-6烷氧基、C 1-6卤代烷基、羟基和氰基；或者R a和R b与相邻碳原子连接成5-6元含氧杂环；
    R c和R d独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基、3-6元杂环基、3-6元杂环基烷基、C 2-6烯基和C 2-6炔基，或者R c和R d与相邻碳原子连接成3-6元的碳环或杂环；
    R e选自氢、C 1-6烷基、C 3-6环烷基、C 1-6卤代烷基、C 1-6烷氧烷基和C 2-C 5杂环基；
    q＝0或1；
    当q＝0时，R c和R d不同时为氢；
    优选地，R a和R b独立选自氢、卤素和C 1-6烷基，或者R a和R b与相邻碳原子连接成5-6元含氧杂环；
    优选地，R a和R b独立选自氢、氟、氯和甲基，或者R a和R b和与其相连的苯环共同形成
    Figure PCTCN2022074328-appb-100011
    其中Z选自-CH 2-、-CD 2-、-CH 2CH 2-和-CF 2-；
    优选地，R a为甲基，R b为氟，或者R a和R b和与其相连的苯环共同形成
    Figure PCTCN2022074328-appb-100012
    优选地，R c和R d独立选自氢、
    Figure PCTCN2022074328-appb-100013
    C 1-6烷氧基烷基和C 1-6烷胺基烷基，或者R c和R d与相邻碳原子连接成3-6元碳环；
    优选地，R c为氢，R d选自氢、
    Figure PCTCN2022074328-appb-100014
    甲氧基乙基和环丙基，或者R c和R d与相邻碳原子连接成3-6元碳环；
    优选地，R e选自氢和C 1-6烷基。
  6. 权利要求1所述的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，其中所述化合物具有式(V)的结构：
    Figure PCTCN2022074328-appb-100015
    上式(V)中，R选自C 3-6环烷基和C 1-6烷氧基；
    A”’选自
    Figure PCTCN2022074328-appb-100016
    中的一种；
    R x”’选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧烷基、C 3- 6环烷基和3-6元杂环基(例如3-6元碳环或3-6元杂环)；
    R y”’和R z”’独立选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基、3-6元杂环基、3-6元杂环基烷基、C 2-6烯基、C 2-6炔基、芳基和杂芳基，或者R y”’和R z”’与相邻碳原子连接成3-6元环；
    优选地，R选自甲基、甲氧基和环丙基；优选地，R y”’和R z”’各自独立地选自氢、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基烷基、C 1-6烷胺基烷基、C 3-6环烷基和乙烯基，或者R y”’和R z”’与相邻碳原子连接成3-6元碳环；
    优选地，R y”’和R z”’均为氢，或者R y”’和R z”’与相邻碳原子连接成3-6元碳环；
    优选地，R x”’选自氢和C 1-6烷基；
    优选地，R x”’为氢。
  7. 权利要求1所述的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，所述化合物结构如下所示：
    Figure PCTCN2022074328-appb-100017
    Figure PCTCN2022074328-appb-100018
    Figure PCTCN2022074328-appb-100019
    Figure PCTCN2022074328-appb-100020
    Figure PCTCN2022074328-appb-100021
    Figure PCTCN2022074328-appb-100022
  8. 式(VI)所示化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，其中所述化合物结构如下所示：
    M-L-E-D
    式(VI)
    其中，
    M是和抗体或其抗原结合片段的接头部位；
    L是连接接头M和E之间的连接子；
    E是连接L和D的结构片段；
    D是细胞毒性药物的结构片段；
    优选地，M选自以下结构：
    Figure PCTCN2022074328-appb-100023
    优选地，M选自以下结构：
    Figure PCTCN2022074328-appb-100024
    优选地，L选自由下述的一个或多个组成的二价结构：C 1-6亚烷基、-N(R’)-、羰基、-O-、Val、Cit、Phe、Lys、D-Val、Leu、Gly、Ala、Asn、Val-Cit、Val-Ala、Val-Lys、Val-Lys(Ac)、Phe-Lys、Phe-Lys(Ac)、D-Val-Leu-Lys、Gly-Gly-Arg、Ala-Ala-Asn、Ala-Ala-Ala、Val-Lys-Ala、Gly-Gly-Gly、Gly-Gly-Phe-Gly、Gly-Gly-Gly-Gly-Gly、
    Figure PCTCN2022074328-appb-100025
    Figure PCTCN2022074328-appb-100026
    Figure PCTCN2022074328-appb-100027
    其中R’代表氢、C 1-6烷基或含-(CH 2CH 2O) r-的烷基；r选自1-10的整数；s选自1-10的整数；
    优选地，L选自以下结构：
    Figure PCTCN2022074328-appb-100028
    优选地，L选自以下结构：
    Figure PCTCN2022074328-appb-100029
    优选地，E选自单键、-NH-CH 2-、
    Figure PCTCN2022074328-appb-100030
    优选地，E为-NH-CH 2-；
    优选地，所述细胞毒性药物选自权利要求1-6任意一项所述的-化合物；
    优选地，所述细胞毒性药物选自权利要求7所述的化合物1-1至1-15；2-1至2-27；3-1至3-26；4-1至4-15；或5-1至5-36；
    优选地，D选自权利要求1-6任意一项所述的-化合物去氢之后的结构；
    优选地，D选自权利要求7所述的化合物1-1至1-15；2-1至2-27；3-1至3-26；4-1至4-15；或5-1至5-36去氢之后的结构；
    优选地，D选自以下结构：
    Figure PCTCN2022074328-appb-100031
    优选地，D优选自以下结构：
    Figure PCTCN2022074328-appb-100032
  9. 根据权利要求8所述的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药，所述化合物结构如下所示：
    Figure PCTCN2022074328-appb-100033
    Figure PCTCN2022074328-appb-100034
    Figure PCTCN2022074328-appb-100035
    Figure PCTCN2022074328-appb-100036
    Figure PCTCN2022074328-appb-100037
    优选地，所述化合物选自：
    Figure PCTCN2022074328-appb-100038
  10. 一种药物组合物，其包含权利要求1-9任一项的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物或前药，以及一种或多种药学上可接受的载体。
  11. 一种药盒产品，其包含：
    a)作为第一治疗剂的至少一种权利要求1-9任一项的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物或前药，或者权利要求10的药物组合物；
    b)任选存在的作为第二治疗剂的至少一种其他治疗剂，或者作为第二药物组合物的包含其他治疗剂的药物组合物；和
    c)任选存在的包装和/或说明书。
  12. 权利要求1-9任一项的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药、权利要求10的药物组合物或者权利要求11的药盒产品在制备用于治疗细胞异常增殖方面的疾病的药物中的用途。
  13. 一种治疗细胞异常增殖方面的疾病的方法，其包括以下步骤：将治疗有效量的权利要求1-9任一项的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮 氧化物、同位素标记物、代谢物和前药或者权利要求10的药物组合物施用于对其有需求的个体向。
  14. 权利要求1-9任一项的化合物或其可药用盐、酯、立体异构体、多晶型物、溶剂合物、氮氧化物、同位素标记物、代谢物和前药、权利要求10的药物组合物或者权利要求11的药盒产品，其用于治疗细胞异常增殖方面的疾病。
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WO2024091437A1 (en) * 2022-10-25 2024-05-02 Merck Sharp & Dohme Llc Exatecan-derived adc linker-payloads, pharmaceutical compositions, and uses thereof

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