Condensed ring compound as well as preparation method and application thereof

Abstract

The invention discloses a fused ring compound as well as a preparation method and application thereof. The fused ring compound has an excellent treatment effect on solid tumors such as gastric cancer, breast cancer, lung cancer or urothelial cancer or malignant tumors of a blood system.

Description

Condensed ring compound and its preparation method and use

[0001] This application is based on the application with CN application number 202310538792.7 and application date May 12, 2023, and claims its priority. The disclosed content of the CN application is hereby introduced as a whole into this application. Technical Field

[0002] The present application relates to the field of medicine, and in particular to a fused ring compound and a preparation method and use thereof. Background Art

[0003] Antibody drug conjugates (ADCs) typically consist of a monoclonal antibody, a bioactive molecule (primarily a tumor-killing cytotoxin), and a linker. The bioactive molecule is covalently coupled to the antibody via the linker. The antibody recognizes specific targets on the surface of tumor cells, guiding the ADC to the tumor microenvironment and the surface of cancer cells, where it enters the cancer cells through endocytosis. The bioactive molecule is then released inside the cancer cells and kills them by inhibiting their microtubules or damaging their DNA, minimizing damage to normal tissue cells.

[0004] Bioactive molecules suitable for ADCs mainly include anti-tubulin agents, DNA intercalators, DNA topoisomerase inhibitors, RNA polymerase inhibitors, and gene transcription inhibitors. Currently, the toxins used in ADCs sold on the market and in clinical trials are mainly microtubule inhibitors, such as MMAE, MMAF, DM1, and DM4.

[0005] The present invention aims to provide a novel fused-ring compound with excellent anti-tumor activity and is suitable for preparing antibody-drug conjugates (ADCs). The conjugates have good stability, homogeneity, hydrophilicity, efficacy, and safety.

[0006] Summary of the Invention

[0007] Compound

[0008] In a first aspect, the present invention provides a compound or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled compound, metabolite or prodrug thereof, wherein the compound has the structure shown in the following formula:

[0009] Among them, R 1 and R 2 are each independently selected from hydrogen, -CN, halogen, -OR a 、-NH2、-NH(C 1-6 Alkyl), -N(C1-6 Alkyl)2, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl groups are optionally substituted by one or more selected from -CN, halogen, -OH, -NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 substituted by aryl and 5-10 membered heteroaryl substituents;

[0010] R 4 and R 4’ are independently selected from hydrogen, halogen, -OH, -NH2, C 1-6 Alkyl, halogenated C 1-6 Alkyl and C 1-6 Alkoxy, R 5 and R 5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms form a carbon-carbon double bond or a 3-6 membered carbon ring, R 5 and R 5’ are all hydrogen; or, R 4 and R 5 Connected to form a bond or together with the connection R 4 and R 5 The carbon atoms form a 3-6 membered carbon ring, R 5’ and R 4’ are independently selected from hydrogen, halogen, -NH2, C 1-6 Alkyl, halogenated C 1-6 Alkyl and C 1-6 alkoxy;

[0011] R 6 For hydrogen, C 1-6 Alkyl or

[0012] R 6’ 、R 7 and R 7’ are all hydrogen; or, R 6’ With R 7 Connected into a bond, R 7’ is hydrogen;

[0013] R aSelected from H, C 1-6 Alkyl or C 3-6 Cycloalkyl;

[0014] Ring A is selected from 3-6 membered cycloalkyl, C 6-10 Aryl, 5-10 membered heteroaryl, 3-6 membered heterocyclic group;

[0015] R 3 Selected from hydrogen, -CN, halogen, -OR a 、-NH2、-NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl groups are optionally substituted by one or more selected from -CN, halogen, -OH, -NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 substituted by aryl and 5-10 membered heteroaryl substituents;

[0016] X and Y are each independently -CH2-, -CD2-, and -C(O)-;

[0017] m is selected from 1-5, for example 1, 2, 3, 4 or 5;

[0018] n is selected from 1-10, for example 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0019] When n is 5 and ring A is phenyl, R 3 Not a methoxy group.

[0020] In some embodiments, R 1 Selected from hydrogen, -CN, halogen, -OR a and C 1-6 Alkyl, where R a Selected from C 1-6 Alkyl, deuterated C 1-6 Alkyl and C 3-6 Cycloalkyl.

[0021] In some embodiments, R 1 Select from -OR a , where R a Selected from C 1-6 Alkyl, deuterated C 1-6 Alkyl and C 3-6 Cycloalkyl.

[0022] In some embodiments, R 1 is selected from methoxy, trideuteromethoxy and -O-cyclopropyl.

[0023] In some embodiments, R 1 Selected from hydrogen, -CN, halogen, -OR a and C 1-6 Alkyl, where R a C 1-6 Alkyl or C 3-6 Cycloalkyl.

[0024] In some embodiments, R 1 For-OR a , where R a C 1-6 Alkyl or C 3-6 Cycloalkyl.

[0025] In some embodiments, R 1 is methoxy or -O-cyclopropyl.

[0026] In some embodiments, R 2 Selected from hydrogen, -CN, halogen, -OR a and C 1-6 Alkyl, where R a C 1-6 Alkyl or C 3-6 Cycloalkyl.

[0027] In some embodiments, R 2 For-OR a , where R a C 1-6 alkyl.

[0028] In some embodiments, R 2 It is a methoxy group.

[0029] In some embodiments, R 4 is hydrogen or halogen, R 4’ Selected from hydrogen, halogen, -OH, -NH2, C 1-6 Alkyl, halogenated C 1-6 Alkyl and C 1- 6 alkoxy, R 5 and R 5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms form a carbon-carbon double bond or a 3-6 membered carbon ring, R 5 and R 5’ are all hydrogen; or, R 4 With R 5Connected to form a bond or together with the connection R 4 and R 5 The carbon atoms form a 3-6 membered carbon ring, R 5’ and R 4’ All are hydrogen.

[0030] In some embodiments, R 4 is hydrogen or fluorine, R 4’ is selected from hydrogen, fluorine, -OH, -NH2, methyl, trifluoromethyl and methoxy, R 5 and R 5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms form a carbon-carbon double bond or a 3-6 membered carbon ring, R 5 and R 5’ are all hydrogen; or, R 4 With R 5 Connected to form a bond or together with the connection R 4 and R 5 The carbon atoms form a 3-membered carbon ring, R 5’ and R 4’ All are hydrogen.

[0031] In some embodiments, R 4 is hydrogen or fluorine, R 4’ is selected from hydrogen, fluorine, -OH, -NH2, methyl, trifluoromethyl and methoxy, R 5 and R 5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms form a carbon-carbon double bond, R 5 and R 5’ are all hydrogen; or, R 4 With R 5 Connected to form a bond or together with the connection R 4 and R 5 The carbon atoms form a 3-membered carbon ring, R 5’ and R 4’ All are hydrogen.

[0032] In some embodiments, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms form a 3-membered carbon ring, R 5 and R 5’ All are hydrogen.

[0033] In some embodiments, R 6 for

[0034] Ring A is selected from 3-6 membered cycloalkyl, C 6-10 Aryl or 5-10 membered heteroaryl;

[0035] R 3 Selected from hydrogen, -CN, halogen, -OR a 、-NH2、-NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl and heteroaryl groups are optionally substituted by one or more selected from -CN, halogen, -OH, -NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 substituted by aryl and 5-10 membered heteroaryl substituents;

[0036] R a Selected from H and C 1-6 alkyl;

[0037] m is selected from 1, 2, 3, 4 and 5.

[0038] In some embodiments, R 6 for

[0039] Ring A is selected from cyclopropyl, phenyl, furyl, thienyl, pyrimidinyl and pyridinyl;

[0040] R 3 Selected from hydrogen, fluorine, -OH, methoxy, -NH2 and hydroxymethyl;

[0041] m is 1 or 2.

[0042] In some embodiments, R 6 for

[0043] Ring A is phenyl;

[0044] R 3 Selected from hydrogen, fluorine, -OH, methoxy, -NH2 and hydroxymethyl;

[0045] m is 1 or 2.

[0046] In some embodiments, Ring A is C 6-10 aryl or 5-10 membered heteroaryl.

[0047] In some embodiments, Ring A is selected from phenyl, pyrimidinyl, pyridinyl, furanyl, and thienyl.

[0048] In some embodiments, R 3 Selected from -OH, methoxy, -NH2 and hydroxymethyl.

[0049] In some embodiments, m is 1 or 2.

[0050] In some embodiments, m is 1.

[0051] In some embodiments, R 6 Selected from phenyl, pyridyl,

[0052] In some embodiments, R 6 Selected from phenyl, pyridyl,

[0053] R 6’ With R 7 Connected into a bond, R 7’ For hydrogen.

[0054] In some embodiments, R 6 for

[0055] R 6’ 、R 7 and R 7’ All are hydrogen.

[0056] In some embodiments, n is 3-8, eg, 3, 4, 5, 6, 7, or 8.

[0057] In some embodiments, X is -CH2- or -CD2-; optionally, Y is selected from -CH2-, -CD2-, and -C(O)-.

[0058] In some embodiments, X is -CH2- and Y is -CH2-; X is -CH2- and Y is -CD2- or -C(O)-; or X is -CD2- and Y is -CH2-.

[0059] In some embodiments, X is -CH2- and Y is -CH2-.

[0060] In some embodiments, the compound has a structure represented by formula (I-1) or formula (I-2):

[0061] Among them, R 1 、R 2 、R 3、R 4 、R 4’ 、R 5 、R 5’ , Ring A, m and n are as described above.

[0062] In some embodiments, the compound has a structure shown in Formula (II-1) or Formula (II-2):

[0063] Among them, R 1 、R 2 、R 3 、R 4 、R 4’ 、R 5 、R 5’ , Ring A, m and n are as described above.

[0064] In some embodiments, in any of the above formulas,

[0065] R 1 is selected from methoxy, trideuteromethoxy and -O-cyclopropyl;

[0066] R 2 is methoxy;

[0067] R 4 is hydrogen or fluorine, R 4’ is selected from hydrogen, fluorine, -OH, -NH2, trifluoromethyl and methoxy, R 5 and R 5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms form a carbon-carbon double bond or a 3-membered carbon ring, R 5 and R 5’ are all hydrogen; or, R 4 With R 5 Connected to form a bond or together with the connection R 4 and R 5 The carbon atoms form a 3-membered carbon ring, R 5’ and R 4’ All are hydrogen;

[0068] Ring A is selected from cyclopropyl, phenyl, furyl, thienyl, pyrimidinyl and pyridinyl;

[0069] R 3 Selected from hydrogen, fluorine, -OH, methoxy, -NH2 and hydroxymethyl;

[0070] m is 1 or 2;

[0071] n is 5.

[0072] In some embodiments, in any of the above formulas,

[0073] R 1 and R 2 All are methoxy;

[0074] R 4 is hydrogen or fluorine, R 4’ is selected from hydrogen, fluorine, -OH, -NH2, trifluoromethyl and methoxy, R 5 and R 5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms form a carbon-carbon double bond or a 3-membered carbon ring, R 5 and R 5’ are all hydrogen; or, R 4 With R 5 Connected to form a bond or together with the connection R 4 and R 5 The carbon atoms form a 3-membered carbon ring, R 5’ and R 4’ All are hydrogen;

[0075] Selected from

[0076] n is 5.

[0077] In some embodiments, the compound is selected from the following structures:

[0078] Antibody Drug Conjugates

[0079] The fused ring compounds of the present invention have excellent anti-tumor activity and are suitable for preparing antibody-drug conjugates. In some embodiments, the resulting antibody-drug conjugates have good stability, homogeneity, hydrophilicity, efficacy, and safety.

[0080] Therefore, in another aspect, the present application provides an antibody drug conjugate containing any one of the compounds described above or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug fragment thereof.

[0081] Pharmaceutical composition

[0082] The second aspect of the present invention provides a pharmaceutical composition comprising the compound described above or its pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope labels, metabolites or prodrugs, or antibody drug conjugates and one or more pharmaceutical excipients.

[0083] The compounds or antibody drug conjugates described herein are typically formulated in a unit injectable form together with a pharmaceutically acceptable parenteral vehicle for parenteral use, such as bolus injection, intravenous injection, intratumoral injection, etc. Optionally, the antibody drug conjugate having the desired purity is mixed with a pharmaceutically acceptable diluent, carrier, excipient or stabilizer in the form of a lyophilized agent or solution (Remington's Pharmaceutical Sciences (1980) 16 th Administration of the compounds or antibody drug conjugates described herein, or pharmaceutical compositions containing the same, can be by any route appropriate to the subject to be treated.

[0084] application

[0085] A third aspect of the present invention provides the use of the aforementioned compound or its pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope-labeled substances, metabolites or prodrugs, the antibody-drug conjugates, or the pharmaceutical compositions in the preparation of drugs for treating cancer or tumors.

[0086] The fourth aspect of the present invention provides the compound described above or its pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug, the antibody drug conjugate, or the pharmaceutical composition, for use in treating cancer or tumors.

[0087] A fifth aspect of the present invention provides a method for treating cancer or tumors, comprising administering to an individual in need thereof a therapeutically effective amount of the compound described above or its pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, nitrogen oxides, isotope labels, metabolites or prodrugs, the antibody drug conjugate, or the pharmaceutical composition.

[0088] In some embodiments of the present invention, the cancer or tumor is a solid tumor or a hematological malignancy; for example, selected from gastric cancer, breast cancer, lung cancer (eg, non-small cell lung cancer, particularly lung adenocarcinoma) and urothelial carcinoma.

[0089] In some embodiments, the compound or pharmaceutical composition thereof is sufficient (e.g., in a subject):

[0090] (1) Inhibit the proliferation of cells (such as tumor cells);

[0091] (2) inhibit tumor growth; or

[0092] (3) induce and / or increase antibody-dependent cellular cytotoxicity activity;

[0093] (4) Any combination of (1) to (3) above.

[0094] definition

[0095] Unless otherwise defined below, the meanings of all technical and scientific terms used herein are intended to be the same as those generally understood by those skilled in the art. Reference to the techniques used herein is intended to refer to techniques generally understood in the art, including variations of those techniques that are obvious to those skilled in the art or replacements with equivalent techniques. Furthermore, laboratory procedures such as genomics, nucleic acid chemistry, and molecular biology used herein are conventional procedures widely used in the corresponding fields. Although it is believed that the following terms are well understood by those skilled in the art, the following definitions are still set forth to better explain the present invention.

[0096] The term "antibody" refers to an immunoglobulin molecule typically composed of two pairs of polypeptide chains, each pair having one light chain (LC) and one heavy chain (HC). Antibody light chains can be classified as kappa (κ) and lambda (λ) light chains. Heavy chains can be classified as μ, δ, γ, α, or ε, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively. Within light and heavy chains, the variable and constant regions are connected by a "J" region of approximately 12 or more amino acids, with heavy chains also containing a "D" region of approximately 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region is composed of three domains: CH1, CH2, and CH3. Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region is composed of one domain, CL. The constant domains are not directly involved in the binding of antibodies to antigens, but exhibit a variety of effector functions, such as mediating the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The VH and VL regions can also be subdivided into regions of high variability, called complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions (VH and VL) of each heavy chain / light chain pair form the antigen binding site, respectively. The allocation of amino acids in each region or domain can follow various numbering systems known in the art.

[0097] The term "complementarity determining region" or "CDR" refers to the amino acid residues in the variable region of an antibody that are responsible for antigen binding. The variable regions of the heavy and light chains each contain three CDRs, designated CDR1, CDR2, and CDR3. The precise boundaries of these CDRs can be defined according to various numbering systems known in the art, for example, as defined in the Kabat numbering system (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), the Chothia numbering system (Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883), the IMGT numbering system (Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003), or the AbM numbering system (Martin ACR, Cheetham JC, Rees AR (1989) Modelling antibody hypervariable loops: A combined algorithm. Proc Natl Acad Sci USA 86:9268-9272). For a given antibody, a person skilled in the art will readily identify the CDRs defined by each numbering system. Furthermore, the correspondence between different numbering systems is well known to those skilled in the art (e.g., see Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003).

[0098] In the present invention, the CDRs contained in an antibody or antigen-binding fragment thereof can be determined according to various numbering systems known in the art, such as those defined by the Kabat, Chothia, IMGT, or AbM numbering systems. In certain embodiments, the CDRs contained in an antibody or antigen-binding fragment thereof are defined by the Chothia numbering system.

[0099] The term "framework region" or "FR" residues refers to those amino acid residues in an antibody variable region other than the CDR residues as defined above.

[0100] The term "antigen-binding fragment" of an antibody refers to polypeptides that are fragments of an antibody, such as polypeptides that are fragments of a full-length antibody, which retain the ability to specifically bind to the same antigen bound by the full-length antibody and / or compete with the full-length antibody for specific binding to the antigen, and are also referred to as "antigen-binding portions." See generally, Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nd ed., Raven Press, NY (1989), which is incorporated herein by reference in its entirety for all purposes. Antigen-binding fragments of antibodies can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. Non-limiting examples of antigen-binding fragments include Fab fragments, Fab' fragments, F(ab)'2 fragments, F(ab)'3 fragments, Fd, Fv, scFv, di-scFv, (scFv)2, disulfide-stabilized Fv proteins ("dsFv"), single domain antibodies (sdAbs, nanobodies), and polypeptides that comprise at least a portion of an antibody sufficient to confer specific antigen-binding ability on the polypeptide. Engineered antibody variants are reviewed in Holliger et al., 2005; Nat Biotechnol, 23: 1126-1136.

[0101] The term "Fd" means an antibody fragment consisting of the VH and CH1 domains; the term "dAb fragment" means an antibody fragment consisting of the VH domain (Ward et al., Nature 341:544-546 (1989)); the term "Fab fragment" means an antibody fragment consisting of the VL, VH, CL and CH1 domains; the term "F(ab')2 fragment" means an antibody fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; the term "Fab' fragment" means a fragment obtained after reducing the disulfide bonds linking the two heavy chain fragments in the F(ab')2 fragment, consisting of one complete light chain and the Fd fragment (consisting of the VH and CH1 domains) of the heavy chain.

[0102] The term "Fv" refers to an antibody fragment consisting of the VL and VH domains of a single arm of an antibody. The Fv fragment is generally considered to be the smallest antibody fragment that can form a complete antigen-binding site. It is generally believed that the six CDRs confer antigen-binding specificity to an antibody. However, even a single variable region (e.g., an Fd fragment, which contains only three CDRs specific for an antigen) can recognize and bind to an antigen, although its affinity may be lower than that of the complete binding site.

[0103] The term "Fc" refers to an antibody fragment formed by disulfide bonds between the second and third constant regions of the first heavy chain and the second and third constant regions of the second heavy chain. The Fc fragment of an antibody has various functions but is not involved in antigen binding.

[0104] The term "scFv" refers to a single polypeptide chain comprising a VL and VH domain, wherein the VL and VH are connected by a linker (see, e.g., Bird et al., Science 242: 423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85: 5879-5883 (1988); and Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, Roseburg and Moore, eds., Springer-Verlag, New York, pp. 269-315 (1994)). Such scFv molecules may have the general structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. Suitable prior art linkers consist of repeated GGGGS amino acid sequences or variants thereof. For example, a linker having the amino acid sequence (GGGGS)4 can be used, but variants thereof can also be used (Holliger et al. (1993), Proc. Natl. Acad. Sci. USA 90:6444-6448). Other linkers useful in the present invention are described by Alfthan et al. (1995), Protein Eng. 8:725-731, Choi et al. (2001), Eur. J. Immunol. 31:94-106, Hu et al. (1996), Cancer Res. 56:3055-3061, Kipriyanov et al. (1999), J. Mol. Biol. 293:41-56, and Roovers et al. (2001), Cancer Immunol. In some cases, a disulfide bond may also be present between the VH and VL of the scFv. In certain embodiments, the VH and VL domains may be positioned relative to each other in any suitable arrangement. For example, containing NH2-VH-VH-COOH, NH 2- VL-VL-COOH scFv.

[0105] The term "single-domain antibody (sdAb)" has the meaning generally understood by those skilled in the art, and refers to an antibody fragment composed of a single monomeric variable antibody domain (e.g., a single heavy chain variable region) that retains the ability to specifically bind to the same antigen as the full-length antibody (Holt, L. et al., Trends in Biotechnology, 21(11):484-490, 2003). Single-domain antibodies are also called nanobodies.

[0106] Each of the above antibody fragments retains the ability to specifically bind to the same antigen as the full-length antibody, and / or competes with the full-length antibody for specific binding to the antigen.

[0107] Herein, unless the context clearly indicates otherwise, when referring to the term "antibody", it includes not only intact antibodies, but also antigen-binding fragments of antibodies.

[0108] Antigen-binding fragments of antibodies (e.g., those described above) can be obtained from a given antibody (e.g., an antibody provided herein) using conventional techniques known to those skilled in the art (e.g., recombinant DNA technology or enzymatic or chemical cleavage methods), and the antigen-binding fragments of antibodies can be screened for specificity in the same manner as for intact antibodies.

[0109] The twenty conventional amino acids referred to herein are denoted according to conventional usage. See, for example, Immunology—A Synthesis (2nd Edition, E.S. Golub and D.R. Gren, Eds., Sinauer Associates, Sunderland, Mass. (1991)), which is incorporated herein by reference. In the present invention, amino acids are generally represented by single-letter and three-letter abbreviations known in the art. For example, alanine can be represented by A or Ala.

[0110] The term "linker" refers to a structural fragment that connects a cytotoxic drug to an antibody or antigen-binding fragment.

[0111] The term "drug-linker" refers to the structure of the cytotoxic drug and linker described herein before being linked to the antibody or antigen-binding fragment thereof. The drug-linker is covalently linked to the antibody or antigen-binding fragment thereof to form the conjugate described herein.

[0112] The "drug-linker" also includes all pharmaceutically acceptable isotope-labeled compounds thereof, which are identical to the "drug-linker" compounds of the present invention except that one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature. Examples of isotopes suitable for inclusion in the present invention include, but are not limited to, isotopes of hydrogen (e.g., 2 H. 3 H, deuterium D, tritium T); carbon isotopes (such as 11 C. 13 C and 14 C); isotopes of chlorine (e.g. 37 Cl); isotopes of fluorine (e.g. 18 F); isotopes of iodine (such as 123 I and 125I); isotopes of nitrogen (e.g. 13 N and 15 N); oxygen isotopes (e.g. 15 O. 17 O and 18 O); and sulfur isotopes (e.g. 35 S).

[0113] The terms "comprises," "comprising," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps.

[0114] The term "alkyl" refers to a group obtained by removing one hydrogen atom from a straight-chain or branched hydrocarbon group, for example, "C 1-20 Alkyl", "C 1-10 Alkyl", "C 1-6 Alkyl", "C 1-4 Alkyl", "C 1-3 alkyl”, etc., specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 1,2-dimethylpropyl, etc.

[0115] As used herein, the term "haloalkyl" refers to an alkyl group substituted by one or more (such as 1 to 3) the same or different halogen atoms. 1-4 "Haloalkyl" refers to C 1-4 The alkyl group is substituted by one or more halogen atoms that are the same or different, such as -CF3, -C2F5, -CHF2, -CH2F, -CH2CF3, -CH2Cl or -CH2CH2CF3.

[0116] As used herein, the term "hydroxyalkyl" refers to a group in which a hydrogen atom in an alkyl group is replaced by one or more hydroxyl groups, for example, C 1- 4-hydroxyalkyl or C 1-6 Examples of hydroxyalkyl include, but are not limited to, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, -CH(OH)CH3, and the like.

[0117] As used herein, the term "alkenyl" refers to a straight or branched aliphatic hydrocarbon group having one or more carbon-carbon double bonds. 2-6The term "alkenyl" refers to an alkenyl group having 2 to 6 carbon atoms and one, two or three carbon-carbon double bonds (e.g., ethenyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, 4-methyl-3-pentenyl, etc.), which is optionally substituted with one or more (e.g., 1 to 3) substituents described herein.

[0118] As used herein, the term "alkynyl" refers to a straight or branched aliphatic hydrocarbon group having one or more carbon-carbon triple bonds. 2-6 The term "alkynyl" refers to an alkynyl group having 2 to 6 carbon atoms and one, two or three carbon-carbon triple bonds (e.g., ethynyl, 1-propynyl, 2-propynyl, 2-butynyl, 3-butynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, etc.), which is optionally substituted with one or more (e.g., 1 to 3) substituents described herein.

[0119] As used herein, the term "alkoxy" refers to an alkyl (as defined above) group attached to the remainder of the molecule via an oxygen atom, such as a C 1-8 Alkoxy, C 1-6 Alkoxy, C 1-4 Alkoxy or C 1-3 Alkoxy. C 1-6 Representative examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, and the like, wherein the alkoxy groups are optionally substituted with one or more (such as 1 to 3) identical or different substituents. For example, the term "haloalkoxy" refers to an alkoxy group wherein the hydrogen atoms are substituted with one or more (such as 1 to 3) identical or different halogen atoms.

[0120] As used herein, the term "cycloalkyl" refers to a saturated or unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring group, including but not limited to monocyclic alkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, etc.) and bicyclic alkyl, including spirocyclic, annular (condensed) or bridged ring systems (i.e., spirocyclic alkyl, annular (condensed) alkyl and bridged cycloalkyl, such as bicyclo [1.1.1] pentyl, bicyclo [2.2.1] heptyl, etc.). In the present invention, cycloalkyl is optionally substituted with one or more (such as 1 to 3) identical or different substituents. The carbon atoms on the cycloalkyl are optionally substituted with oxo (oxo) groups (i.e., forming C=O). The term "C 3-8 "Cycloalkyl" refers to a cycloalkyl group having 3 to 8 ring carbon atoms, such as C3-6 Cycloalkyl, which may be a monocyclic alkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl, or a bicyclic alkyl, such as C 5-8 Spiroalkyl, C 5-8 Bridged cycloalkyl, C 5-8 Condensed cycloalkyl, C 5-6 Spiroalkyl, C 5-6 Bridged cycloalkyl or C 5- 6-fused cycloalkyl. The term "aliphatic ring" refers to a saturated or unsaturated carbon ring containing 3 or more carbon atoms connected together, common aliphatic rings include cyclopropane, cyclobutane, cyclopentane, cyclohexane, etc.

[0121] As used herein, the term "heterocyclyl" or "heterocycle" refers to an aliphatic, saturated or partially unsaturated monocyclic or polycyclic (e.g., cyclic, spirocyclic or bridged) group having 2 or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14) carbon atoms and one or more (e.g., 1, 2, 3 or 4) heteroatoms, including but not limited to oxygen atoms, nitrogen atoms and sulfur atoms, wherein the carbon atoms and heteroatoms on the heterocyclyl are optionally substituted with oxo groups (e.g., forming C=O, S(=O) or S(=O)2), or are optionally substituted with one or more (e.g., 1 to 3) independently selected from halogen and C 1-3 The alkyl group is substituted with a substituent.

[0122] As used herein, the term "aryl" or "aromatic ring" refers to an all-carbon monocyclic or fused polycyclic aromatic group having a conjugated π electron system. 6-10 "Aryl" or "aromatic ring" refers to an aromatic group or aromatic ring containing 6 to 10 carbon atoms, for example, phenyl or a benzene ring, naphthyl or a naphthalene ring. The aryl group is optionally substituted by one or more (such as 1 to 3) identical or different substituents (for example, halogen, OH, CN, NO2, C 1- 6 alkyl, etc.) substituted.

[0123] As used herein, the term "heteroaryl" or "heteroaromatic ring" refers to a monocyclic or polycyclic aromatic group containing one or more identical or different heteroatoms, including monocyclic heteroaryl groups and bicyclic or polycyclic ring systems containing at least one heteroaromatic ring (an aromatic ring system containing at least one heteroatom), which can have 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, for example 5, 6, 7, 8, 9 or 10 ring atoms. The heteroatom can be oxygen, nitrogen or sulfur. The carbon atoms and heteroatoms on the heteroaryl are optionally substituted with oxo groups (for example, to form C=O, S(=O) or S(=O)2).

[0124] As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

[0125] The term "isotopically labeled compound" means a compound that is identical in structure to a compound of the present invention except that one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature. Examples of suitable isotopes for inclusion in the present invention include, but are not limited to, isotopes of hydrogen (e.g., 2 H. 3 H, deuterium D, tritium T); carbon isotopes (such as 11 C. 13 C and 14 C); isotopes of chlorine (e.g. 37 Cl); isotopes of fluorine (e.g. 18 F); isotopes of iodine (such as 123 I and 125 I); isotopes of nitrogen (e.g. 13 N and 15 N); oxygen isotopes (e.g. 15 O. 17 O and 18 O); and sulfur isotopes (e.g. 35 S).

[0126] The term "substituted" refers to the replacement of one or more (e.g., 1, 2, 3, 4, or 5) hydrogen atoms on a designated compound or structural fragment with a substituent, provided that the normal valence of the designated atom in the present context is not exceeded and the substitution forms a stable compound. Combinations of substituents and / or variables are permitted only if such combinations form stable compounds. For example, the substituents are each independently composed of one or more of the following structures: -O-, -S-, -NR'-, halogen, -CN, -OH, -NH2, -NO2, -CN, =O, C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkylene, 3-8 membered heterocyclyl, C6-C 10 (Ethyl)aryl and 5-10 membered (Ethyl)heteroaryl, etc.

[0127] If substituents are described as being "independently selected" from a group, each substituent is selected independently of the other. Thus, each substituent may be the same as or different from another (other) substituent.

[0128] As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5, 6, 7, 8, 9 or 10, where reasonable.

[0129] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0130] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In a compound with one or more (e.g., one, two, three, or four) asymmetric centers, it can produce a racemic mixture, a single enantiomer, a diastereomeric mixture, and a separate diastereomer. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. For example, nitroso-oxime can exist in the following tautomeric form equilibrium in solution:

[0131] It is to be understood that the scope of this application encompasses all such isomers in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%) or mixtures thereof.

[0132] In this article, solid lines (————), solid wedges or virtual wedge The chemical bonds of the compounds of the present invention are depicted. The use of solid lines to depict bonds to asymmetric carbon atoms is intended to indicate that all possible stereoisomers at that carbon atom are included (e.g., specific enantiomers, racemic mixtures, etc.). The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the indicated stereoisomers exist. When present in a racemic mixture, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the present invention are intended to exist as stereoisomers, including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, atropisomers, and mixtures thereof. The compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

[0133] The present invention encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs or mixtures of more than one polymorph in any ratio.

[0134] A cocrystal refers to a drug active molecule and other physiologically acceptable acid, base, salt, or non-ionic compound molecules bound in the same crystal lattice by hydrogen bonds, π-π stacking, van der Waals forces, and other non-covalent bonds.

[0135] It should also be understood that certain compounds of the present invention may be used therapeutically in free form or, where appropriate, in the form of pharmaceutically acceptable derivatives thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, N-oxides, metabolites, or prodrugs that, upon administration to a patient in need thereof, are capable of directly or indirectly providing a compound of the present invention or a metabolite or residue thereof. Therefore, when reference is made herein to a "compound of the present invention," such various derivative forms of the compound are also intended to be encompassed.

[0136] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof, such as hexafluorophosphate salts and meglumine salts. For a review of suitable salts, see Stahl and Wermuth, "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley-VCH, 2002).

[0137] As used herein, the term "ester" refers to esters derived from the compounds of the general formulae herein, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the present invention in the form of free acid or alcohol). The compounds of the present invention themselves may also be esters.

[0138] The compounds of the present invention may exist in the form of solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0139] Those skilled in the art will appreciate that, because nitrogen requires available lone pairs of electrons to be oxidized to oxides, not all nitrogen-containing heterocycles can form N-oxides. Those skilled in the art will recognize nitrogen-containing heterocycles that can form N-oxides. Those skilled in the art will also recognize that tertiary amines can form N-oxides. The synthetic method for preparing the N-oxide of heterocycles and tertiary amines is well known to those skilled in the art, including but not limited to oxidizing heterocycles and tertiary amines with peroxyacids such as Peracetic Acid and Metachloroperbenzoic Acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate and dioxirane such as dimethyldioxirane. These methods for preparing N-oxides have been extensively described and reviewed in the literature, see for example: TL Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp 748-750; AR Katritzky and AJ Boulton, Eds., Academic Press; and GWH Cheeseman and ESGWerstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp 390-392, AR Katritzky and AJ Boulton, Eds., Academic Press.

[0140] Also included within the scope of the present invention are metabolites of the compounds of the invention, i.e., substances formed in vivo upon administration of the compounds of the invention. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc. of the administered compound. Thus, the present invention includes metabolites of the compounds of the invention, including compounds produced by contacting a compound of the invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

[0141] The present invention further includes within its scope prodrugs of the compounds of the present invention, which are certain derivatives of the compounds of the present invention that may themselves have little or no pharmacological activity and can be converted into compounds of the present invention having the desired activity by, for example, hydrolytic cleavage when administered to the body or thereon. Typically, such prodrugs will be functional group derivatives of the compounds that are readily converted into the desired therapeutically active compounds in vivo. Additional information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems", Volume 14, ACS Symposium Series (T. Higuchi and V. Stella). Prodrugs of the present invention can be prepared, for example, by replacing appropriate functional groups present in the compounds of the present invention with certain moieties known to those skilled in the art as "pro-moieties" (e.g., as described in "Design of Prodrugs", H. Bundgaard (Elsevier, 1985)).

[0142] The present invention also encompasses compounds of the present invention that contain protecting groups. During 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 of the molecules involved, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved using conventional protecting groups, for example, those described in TW Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

[0143] If a functional group or structural moiety is described as "substituted or unsubstituted" or "optionally substituted with," the functional group or structural moiety may be (1) unsubstituted or (2) substituted. DETAILED DESCRIPTION

[0144] The present invention will be further described below by describing specific embodiments, but this is not intended to limit the present invention. Those skilled in the art can make various modifications or improvements based on the teachings of the present invention without departing from the basic idea and scope of the present invention.

[0145] The structures of the compounds described in the following examples were determined by nuclear magnetic resonance ( 1 H NMR) or mass spectrometry (MS).

[0146] Nuclear magnetic resonance (1H NMR) measurements were performed using a Bruker 400 MHz NMR spectrometer; the deuterated reagent was hexadeuterated dimethyl sulfoxide (DMSO-d6); and the internal standard was tetramethylsilane (TMS).

[0147] The abbreviations used in the nuclear magnetic resonance (NMR) spectra in the examples are shown below.

[0148] s: singlet, d: doublet, t: triplet, q: quartet, m: multiplet, br: broad, J: coupling constant, Hz: hertz, DMSO-d6: deuterated dimethyl sulfoxide. δ values ​​are expressed in ppm.

[0149] Mass spectrometry (MS) was performed using an Agilent (ESI) mass spectrometer, model Agilent 6120B.

[0150] Preparation Example 1: (S)-8-((5-(((S)-2-(4-aminophenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-5)

[0151] Step 1: Preparation of (S)-8-(benzyloxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one (7-5-2)

[0152] Dissolve (S)-8-(Benzyloxy)-7-methoxy-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5,11(10H)-dione (2 g, 5.29 mmol, FR) in tetrahydrofuran (100 mL), add sodium borohydride (599.85 mg, 15.86 mmol), and then add trifluoroacetic acid (1.21 g, 10.57 mmol) dropwise at room temperature until no more gas escapes. Heat to 75°C and reflux for 5 hours.

[0153] The reaction solution was concentrated, water (50 mL) was added, and the mixture was extracted twice with ethyl acetate (50 mL x 2). The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude title compound (1.82 g, 4.99 mmol), which was used in the next step without purification.

[0154] Its structural characterization data are as follows:

[0155] ESI-MS (m / z): 365.2 [M+H] +

[0156] Step 2: Preparation of (S)-8-hydroxy-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one (7-5-3)

[0157] Dissolve (S)-8-(Benzyloxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one (100 mg, 0.274 mmol, FR) in dichloromethane (0.8 mL) and cool to 0°C. Then, add methanesulfonic acid (0.4 mL) dropwise. Incubate for 1 hour. Add water (2 mL) to the reaction mixture, then adjust the pH to 8 with saturated sodium bicarbonate solution. Extract with dichloromethane (3 mL x 5) five times. The combined organic phases are washed with saturated sodium chloride solution (3 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude title compound (75 mg, 0.273 mmol), which is used directly in the next step without purification.

[0158] Its structural characterization data are as follows:

[0159] ESI-MS (m / z): 275.2 [M+H] +

[0160] Step 3: Preparation of tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxy-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-5-5)

[0161] (S)-8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl trifluoromethanesulfonate (550 mg, 0.987 mmol), 4-(tert-butoxycarbonyl)aminophenylboronic acid (468 mg, 1.97 mmol), and sodium carbonate (418 mg, 3.95 mmol) were dissolved in a mixed solvent of toluene (10 mL), ethanol (1 mL), and water (1 mL). Tetrakis(triphenylphosphine)palladium (114 mg, 0.099 mmol) was added, the atmosphere was replaced with nitrogen twice, and the mixture was heated at 80°C under protection for 3 hours. The reaction mixture was added to ethyl acetate (20 mL) and water (10 mL), stirred, filtered, and the filtrate was allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0% to 50%) and concentrated again to obtain the title compound (397 mg, 0.661 mmol).

[0162] Its structural characterization data are as follows:

[0163] ESI-MS (m / z): 600.7 [M+H] +

[0164] Step 4: Preparation of tert-butyl (4-((S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-5,11-dioxo-5,10,11,11a-tetrahydro-1-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-5-6)

[0165] Tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxy-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (100 mg, 0.167 mmol) and (S)-8-hydroxy-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (46 mg, 0.167 mmol) were dissolved in DMF (1 mL). Potassium carbonate (69 mg, 0.500 mmol) was added and the mixture was stirred for 16 hours. The reaction was quenched by adding water, and the organic phases were combined after extraction with ethyl acetate three times (10 mL x 3), washed with saturated brine, and then dried and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) gave the title compound (130 mg, 0.165 mmol).

[0166] Its structural characterization data are as follows:

[0167] ESI-MS (m / z): 794.4 [M+H] +

[0168] Step 5: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-5-7)

[0169] Tert-butyl (4-((S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-5,11-dioxo-5,10,11,11a-tetrahydro-1-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (200 mg, 0.122 mmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (0.5 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. The residue was concentrated under reduced pressure, purified by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%), and lyophilized to obtain the title compound (31 mg, 0.045 mmol).

[0170] Its structural characterization data are as follows:

[0171] ESI-MS (m / z): 694.4 [M+H]+

[0172] Step 6: Preparation of (S)-8-((5-(((S)-2-(4-aminophenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-5)

[0173] (S)-2-(4-Aminophenyl)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (31 mg, 0.045 mmol) was dissolved in dry tetrahydrofuran (5 mL). Sodium borohydride (8.5 mg, 0.224 mmol) was added, and trifluoroacetic acid (20.4 mg, 0.179 mmol) was slowly added dropwise, resulting in the generation of a large amount of gas. After the gas dissipated, the temperature was slowly raised to 70°C and the reaction was refluxed for 16 hours. The reaction solution was cooled to room temperature, methanol was added dropwise to quench the reaction, and the residue was concentrated under reduced pressure. Purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 50%) and freeze-dried to give the title compound (4.65 mg, 0.007 mmol).

[0174] Its structural characterization data are as follows:

[0175] ESI-MS (m / z): 680.4 [M+H] +

[0176] 1H NMR (400MHz, DMSO): δ7.31(d,J=13.4Hz,2H),7.26(s,1H),7.11(d,J=8.4Hz,2H),6.53(d,J=8.5Hz,3H),6. 29(d,J=8.1Hz,3H),5.19(s,2H),4.13(d,J=4.4Hz,1H),3.93(dd,J=12.0,6.1Hz,4H),3.83(d,J=7.6Hz,1H ),3.65(d,J=7.5Hz,6H),3.56-3.46(m,3H),3.31-3.11(m,4H),2.69(dd,J=16.7,3.9Hz,1H),1.95(dd,J=1 2.5,7.3Hz,1H),1.79(dd,J=12.5,7.4Hz,5H),1.56(d,J=6.8Hz,2H),0.67-0.62(m,1H),0.60-0.50(m,3H).

[0177] The preparation method is as follows:

[0178] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0179] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0180] Preparation Example 2: Preparation of (S)-8-((5-(((S)-2-(4-hydroxyphenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-3)

[0181] Step 1: Preparation of (S)-2-(4-(benzyloxy)phenyl)-8-((5-bromopentyl)oxy)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-3-1)

[0182] (S)-8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl trifluoromethanesulfonate (500 mg, 897.13 μmol), (4-(benzyloxy)phenyl)boronic acid (409.18 mg, 1.79 mmol), tetrakistriphenylphosphine palladium (103.67 mg, 89.71 μmol), and sodium carbonate (380.34 mg, 3.59 mmol) were dissolved in toluene (8 mL), water (2 mL), and ethanol (2 mL). The reaction was allowed to proceed at 80°C for 3 hours. After completion, the mixture was extracted with water and ethyl acetate. The organic phases were combined, dried, and concentrated to yield the crude product. The crude product was purified by silica gel column chromatography to yield the title compound (303 mg, 461.04 μmol).

[0183] Its structural characterization data are as follows:

[0184] ESI-MS (m / z): 591.4 [M+H] +

[0185] Step 2: Preparation of (S)-2-(4-(benzyloxy)phenyl)-8-((5-bromopentyl)oxy)-7-methoxy-10-(2-(trimethylsilyl)ethoxy)methyl)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-3-2)

[0186] (S)-2-(4-(Benzyloxy)phenyl)-8-((5-bromopentyl)oxy)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (186 mg, 314.46 μmol) was dissolved in tetrahydrofuran (5 mL) and cooled to -78°C. Then, n-butyllithium (294.81 μL, 471.69 μmol) was added dropwise. The reaction was allowed to react for 0.5 hours. 2-(Trimethylsilyl)ethoxymethyl chloride (78.64 mg, 471.69 μmol) was added dropwise to the reaction mixture. The reaction was continued at the same temperature for 1 hour. After the reaction was complete, saturated ammonium chloride was added to quench the reaction and the mixture was extracted with ethyl acetate. The organic phase was dried and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give the title compound (200 mg, 263.25 μmol).

[0187] Its structural characterization data are as follows:

[0188] ESI-MS (m / z): 721.3 [M+H] +

[0189] Step 3: Preparation of (S)-2-(4-(benzyloxy)phenyl)-8-((5-bromopentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-3-3)

[0190] Sodium borohydride (52.42 mg, 1.39 mmol) and calcium chloride (153.77 mg, 1.39 mmol) were dissolved in a mixture of tetrahydrofuran (2 mL) and ethanol (2 mL). A solution of (S)-2-(4-(benzyloxy)phenyl)-8-((5-bromopentyl)oxy)-7-methoxy-10-(2-(trimethylsilyl)ethoxy)methyl)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (100 mg, 138.55 μmol) in tetrahydrofuran (2 mL) was slowly added dropwise to the mixture. The reaction was allowed to proceed at room temperature for 16 hours. After completion, formic acid (0.5 mL) was added to quench the reaction. Stirring was continued for 10 minutes. Conversion of the intermediate phase was confirmed to be complete. The reaction solution was purified by preparative method to obtain the title compound (14 mg, 23.03 μmol).

[0191] Its structural characterization data are as follows:

[0192] ESI-MS (m / z): 577.3 [M+H] +

[0193] The preparation method is as follows:

[0194] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0195] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0196] Step 4: Preparation of (S)-8-((5-(((S)-2-(4-(benzyloxy)phenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-3-4)

[0197] (S)-2-(4-(Benzyloxy)phenyl)-8-((5-bromopentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (12 mg, 20.78 μmol) and (S)-8-hydroxy-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (5.70 mg, 20.78 μmol) were dissolved in DMF (2 mL). Potassium carbonate (8.62 mg, 62.34 μmol) was added and the mixture was reacted at room temperature for 4 hours. After completion of the reaction, the mixture was quenched with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried, and concentrated. The crude product was purified by silica gel column chromatography to obtain the title compound (10 mg, 12.32 μmol).

[0198] Its structural characterization data are as follows:

[0199] ESI-MS (m / z): 771.4 [M+H] +

[0200] Step 5: Preparation of (S)-8-((5-(((S)-2-(4-hydroxyphenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-3)

[0201] (S)-8-((5-(((S)-2-(4-(benzyloxy)phenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (10.00 mg, 12.97 μmol) was dissolved in dichloromethane (2 mL). Methanesulfonic acid (0.5 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 16 hours. After completion of the reaction, the reaction solution was preparatively purified and lyophilized to give the title compound (0.47 mg, 0.69 μmol).

[0202] Its structural characterization data are as follows:

[0203] ESI-MS (m / z): 681.3 [M+H] +

[0204] 1 H NMR (400 MHz, DMSO): δ1H NMR(400MHz,DMSO)δ9.49(s,1H),7.37(s,1H),7.32(s,1H),7.29(s,1H),7.2 6(d,J=8.8Hz,2H),6.73(d,J=8.4Hz,2H),6.54(d,J=6.4Hz,1H),6.31-6.28( m,3H),4.20-4.12(m,1H),3.95-3.89(m,3H),3.86-3.81(m,1H),3.66(s,3H) ,3.64(s,3H),3.57-3.46(m,3H),3.30-3.28(m,1H),3.26-3.20(m,1H),3.17 -3.11(m,1H),2.76-2.67(m,2H),1.98-1.93(m,1H),1.83-1.77(m,4H),1. 59-1.52(m,2H),0.67-0.62(m,1H),0.60-0.56(m,1H),0.56-0.50(m,2H).

[0205] The preparation method is as follows:

[0206] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0207] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% ammonium bicarbonate)

[0208] Preparation Example 3: Preparation of (11aS,11a"S)-8,8"-(pentane-1,5-diylbis(oxy))bis(7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo][e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one) (7-9)

[0209] Step 1: Preparation of 8,8"-(pentane-1,5-diylbis(oxy))(11aS,11a"S)-bis(7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylic acid diallyl ester)(7-9-2)

[0210] (S)-8-Hydroxy-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylic acid allyl ester (55 mg, 153.47 μmol) and 1,5-dibromopentane (17.64 mg, 76.73 μmol) were added to DMF (2 mL), followed by potassium carbonate (63.63 mg, 460.40 μmol). The mixture was stirred at room temperature for 20 hours. After completion of the reaction, 10 mL of water was added and the mixture was extracted three times with ethyl acetate (5 mL x 3). The combined organic phases were washed with 5 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (petroleum ether / ethyl acetate = 1:2) and concentrated again under reduced pressure to obtain the title compound (100 mg, 127.41 μmol).

[0211] Its structural characterization data are as follows:

[0212] ESI-MS (m / z): 785.4 [M+H] +

[0213] Step 2: Preparation of (11aS,11a"S)-8,8"-(pentane-1,5-diylbis(oxy))bis(7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo][e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one) (7-9)

[0214] 8,8"-(pentane-1,5-diylbis(oxy))(11aS,11a"S)-bis(7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylic acid diallyl ester) (80 mg, 101.92 μmol) and tetrahydropyrrole (14.50 mg, 203.85 μmol) were dissolved in DMF (1.5 mL). Pd(PPh3)4 (11.78 mg, 10.19 μmol) was added under nitrogen protection. The nitrogen atmosphere was replaced three times, and then the reaction was stirred at room temperature under nitrogen protection for 2 hours. After completion of the reaction, 5 mL of water was added to the reaction solution, and the mixture was extracted with 5 mL of ethyl acetate × 3. The organic phase was washed with 5 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (ethyl acetate:methanol = 20:1) to remove triphenylphosphine and then concentrated under reduced pressure. It was then purified by preparative high-performance liquid chromatography and freeze-dried to obtain the title compound (12.83 mg, 20.18 μmol).

[0215] Its structural characterization data are as follows:

[0216] ESI-MS (m / z): 617.4 [M+H] + ;309.3[M / 2+1] +

[0217] 1 H NMR (400MHz, CDCl3) δ7.54(s,2H),6.10(s,2H),3.99(dd,J=16.0,10.4Hz,3H),3.88(d,J=30.0Hz,3H),3.74-3.38(m,4H),2 .04(dd,J=17.2,7.6Hz,1H),1.96-1.83(m,2H),1.71(ddd,J=21.6,12.0,5.6Hz,2H),0.77-0.66(m,1H),0.66-0.50(m,3H).

[0218] The preparation method is as follows:

[0219] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0220] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0221] Preparation Example 4: Preparation of (S)-8-((5-(((S)-2-(4-aminophenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-(methoxy-d3)-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-10)

[0222] Step 1: Preparation of methyl 4-bromo-5-fluoro-2-nitrobenzoate (7-10-2)

[0223] 4-Bromo-5-fluoro-2-nitrobenzoic acid (5.9 g, 22.35 mmol, FR) was dissolved in methanol (30 mL), and thionyl chloride (30 mL) was added. The mixture was reacted at room temperature for 16 hours. The reaction solution was concentrated to obtain a crude product, which was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0% to 50%) and concentrated again to obtain the title compound (3.05 g, 10.97 mmol).

[0224] Step 2: Preparation of methyl 4-bromo-5-(methoxy-d3)-2-nitrobenzoate (7-10-3)

[0225] Methyl 4-bromo-5-fluoro-2-nitrobenzoate (1.5 g, 5.4 mmol, FR) was dissolved in DMF (6 mL). Deuterated methanol (388.5 mg, 10.8 mmol) and cesium carbonate (2.64 mg, 8.09 mmol) were added. The atmosphere was purged with nitrogen three times and allowed to react at room temperature for 16 hours. The reaction solution was added to ethyl acetate (20 mL) and water (30 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was then purified on a silica gel column (ethyl acetate / petroleum ether = 0% to 50%) and concentrated to obtain the title compound (1.08 g, 3.68 mmol).

[0226] Its structural characterization data are as follows:

[0227] 1 H NMR (400MHz, DMSO): δ8.35(s,1H),7.51(s,1H).

[0228] Step 3: Preparation of methyl 5-(methoxy-d3)-2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (7-10-4)

[0229] Methyl 4-bromo-5-(methoxy-d3)-2-nitrobenzoate (1.08 g, 3.68 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (2.34 g, 9.21 mmol), bistriphenylphosphine palladium dichloride (775.94 mg, 1.11 mmol), and potassium acetate (1.08 g, 11.05 mmol) were added to a single-necked flask and added to 1,4-dioxane (15 mL). The atmosphere was replaced with nitrogen three times, the temperature was raised to 101 ° C. and the reaction was carried out for 16 hours. The mixture was cooled to room temperature, filtered through diatomaceous earth, and concentrated to obtain a crude product (1.1 g, 3.23 mmol), which was used directly in the next step without purification.

[0230] Step 4: Preparation of methyl 4-hydroxy-5-(methoxy-d3)-2-nitrobenzoate (7-10-5)

[0231] Methyl 5-(methoxy-d3)-2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (1.1 g, 3.23 mmol) was dissolved in tetrahydrofuran (30 mL). Glacial acetic acid (3.75 mL) was added at 0°C, and hydrogen peroxide (7 mL) was slowly added dropwise. The reaction mixture was reacted at room temperature for 3 hours. Saturated sodium sulfite solution (30 mL) was added to the reaction solution and stirred for 1 hour. The mixture was concentrated under reduced pressure until no tetrahydrofuran remained. 30 mL of ethyl acetate was added for extraction. After separation, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product, which was purified on a silica gel column (ethyl acetate / petroleum ether = 0% to 70%) and concentrated to obtain the title compound (680 mg, 2.95 mmol).

[0232] Its structural characterization data are as follows:

[0233] ESI-MS (m / z): 230.2 [MH] -

[0234] Step 5: Preparation of methyl 4-(benzyloxy)-5-(methoxy-d3)-2-nitrobenzoate (7-10-6)

[0235] Methyl 4-hydroxy-5-(methoxy-d3)-2-nitrobenzoate (680 mg, 2.95 mmol) was dissolved in DMF (5 mL), and potassium carbonate (815.3 mg, 5.91 mmol) and benzyl bromide (1.01 g, 5.91 mmol) were added. The mixture was reacted at room temperature for 15 hours. After adding 30 mL of water, 30 mL of ethyl acetate was added for extraction. After separation, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product, which was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0% to 50%) to obtain the title compound (688 mg, 2.15 mmol).

[0236] Step 6: Preparation of 4-(benzyloxy)-5-(methoxy-d3)-2-nitrobenzoic acid (7-10-7)

[0237] Methyl 4-(benzyloxy)-5-(methoxy-d3)-2-nitrobenzoate (688 mg, 2.15 mmol) was dissolved in tetrahydrofuran (5 mL) and water (1 mL), and lithium hydroxide (103.1 mg, 4.3 mmol) was added. The mixture was reacted at room temperature for 3 hours. The pH of the system was adjusted to 5 with dilute hydrochloric acid, and a solid precipitated. The solid was extracted with 30 mL of ethyl acetate. After separation, the mixture was washed with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the title compound (400 mg, 1.31 mmol).

[0238] Step 7: Preparation of (S)-5-(4-(benzyloxy)-5-(methoxy-d3)-2-nitrobenzoyl)-5-azaspiro[2.4]heptane-6-carboxylic acid methyl ester (7-10-8)

[0239] 4-(Benzyloxy)-5-(methoxy-d3)-2-nitrobenzoic acid (400 mg, 1.31 mmol) and (S)-5-azaspiro[2.4]heptane-6-carboxylic acid methyl ester (203 mg, 1.31 mmol) were dissolved in DMF (5 mL), and HATU (597.3 mg, 1.57 mmol) and DIPEA (507 mg, 3.93 mmol) were added. The mixture was reacted at room temperature for 15 hours. After adding 30 mL of water, 30 mL of ethyl acetate was added for extraction. After separation, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product, which was purified on a silica gel column (ethyl acetate / petroleum ether = 0% to 50%) and concentrated to obtain the title compound (570 mg, 1.29 mmol).

[0240] Its structural characterization data are as follows:

[0241] ESI-MS (m / z): 444.1 [M+H] +

[0242] Step 8: Preparation of (S)-8-(benzyloxy)-7-(methoxy-d3)-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5,11(10H)-dione (7-10-9)

[0243] Methyl (S)-5-(4-(benzyloxy)-5-(methoxy-d3)-2-nitrobenzoyl)-5-azaspiro[2.4]heptane-6-carboxylate (570 mg, 1.29 mmol) was dissolved in methanol (30 mL), and a 10% aqueous solution of ammonium chloride (4 mL) and zinc powder (1.28 g, 19.51 mmol) were added. The temperature was raised to 40°C for 1 hour, then raised to 80°C for overnight. The crude product was concentrated to obtain a clear product, which was dissolved in 35 mL of ethyl acetate and washed three times with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by reverse phase column chromatography (acetonitrile / 1% aqueous formic acid = 0% to 70%) and lyophilized to obtain the title compound (306 mg, 0.8 mmol).

[0244] Its structural characterization data are as follows:

[0245] ESI-MS (m / z): 382.2 [M+H] +

[0246] Step 9: Preparation of (S)-8-hydroxy-7-(methoxy-d3)-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5,11(10H)-dione (7-10-10)

[0247] Dissolve (S)-8-(Benzyloxy)-7-(methoxy-d3)-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5,11(10H)-dione (132 mg, 0.29 mmol) in dichloromethane (1 mL) and cool to 0°C. Then, add methanesulfonic acid (0.5 mL) dropwise. Incubate for 1 hour. Add water (3 mL) to the reaction mixture, then adjust the pH to 8 with saturated sodium bicarbonate solution. Extract three times with dichloromethane (5 mL x 3). The combined organic phases are washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude title compound (100 mg, 0.27 mmol), which is used directly in the next step without purification.

[0248] Its structural characterization data are as follows:

[0249] ESI-MS (m / z): 292.1 [M+H] +

[0250] Step 10: Preparation of tert-butyl (4-((S)-7-methoxy-8-((5-(((S)-7-(methoxy-d3))-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-5,11-dioxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-10-11)

[0251] Tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxy-50,11,11-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (167 mg, 0.27 mmol) and (S)-8-hydroxy-7-(methoxy-d3)-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5,11(10H)-dione (100 mg, 0.27 mmol) were dissolved in DMF (2 mL). Potassium carbonate (60 mg, 0.54 mmol) was added and the reaction was stirred for 16 hours. The reaction was quenched by adding water, and the organic phases were combined after extraction with ethyl acetate three times (10 mL x 3), washed with saturated brine, and then dried and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) gave the title compound (120 mg, 0.136 mmol).

[0252] Its structural characterization data are as follows:

[0253] ESI-MS (m / z): 797.3 [M+H] +

[0254] Step 11: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((S)-7-(methoxy-d3)-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-10-12)

[0255] Tert-butyl (4-((S)-7-methoxy-8-((5-(((S)-7-(methoxy-d3))-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-5,11-dioxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (120 mg, 0.136 mmol) was dissolved in dichloromethane (3 mL) and trifluoroacetic acid (0.6 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. The residue was concentrated under reduced pressure, purified by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%), and lyophilized to obtain the title compound (35 mg, 0.05 mmol).

[0256] Its structural characterization data are as follows:

[0257] ESI-MS (m / z): 697.3 [M+H] +

[0258] Step 12: Preparation of (S)-8-((5-(((S)-2-(4-aminophenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-(methoxy-d3)-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-10)

[0259] (S)-2-(4-Aminophenyl)-7-methoxy-8-((5-(((S)-7-(methoxy-d3)-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (35 mg, 0.05 mmol) was dissolved in dry tetrahydrofuran (5 mL). Sodium borohydride (8.8 mg, 0.225 mmol) was added, and trifluoroacetic acid (20.9 mg, 0.18 mmol) was slowly added dropwise, resulting in the generation of a large amount of gas. After the gas dissipated, the temperature was slowly raised to 70°C and the reaction was refluxed for 16 hours. The reaction solution was cooled to room temperature, methanol was added dropwise to quench the reaction, and the residue was concentrated under reduced pressure. Purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 50%) and freeze-dried to give the title compound (2.9 mg, 0.004 mmol).

[0260] Its structural characterization data are as follows:

[0261] ESI-MS (m / z): 683.3 [M+H] +

[0262] 1H NMR (400MHz, DMSO): δ7.33(s,1H),7.30(s,1H),7.27(s,1H),7.13(d,J=8.4Hz,1H), 6.58-6.53(m,2H),6.34-6.26(m,2H),4.20-4.10(m,1H),4.0-3.92(m,4H),3.86-3.8 0(m,2H),3.67(s,3H),3.56(s,3H),3.3-3.22(m,3H),3.20-3.12(m,2H),2.76-2.66 (m,2H),2.02-1.94(m,1H),1.85-1.77(m,4H),1.63-1.52(m,2H),0.62-0.52(m,3H).

[0263] The preparation method is as follows:

[0264] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0265] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0266] Preparation Example 5: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((S)-7-cyclopropyloxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-11)

[0267] Step 1: Preparation of methyl 4-bromo-5-cyclopropyloxy-2-nitrobenzoate (7-11-1)

[0268] Methyl 4-bromo-5-fluoro-2-nitrobenzoate (1.5 g, 5.4 mmol, FR) was dissolved in DMF (6 mL). Cyclopropanol (635 mg, 10.97 mmol) and cesium carbonate (2.68 mg, 8.3 mmol) were added. The atmosphere was replaced with nitrogen three times and the mixture was allowed to react at room temperature for 16 hours. The reaction mixture was added to ethyl acetate (30 mL) and water (30 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (ethyl acetate / petroleum ether = 0% to 50%) and concentrated to obtain the title compound (750 mg, 2.37 mmol).

[0269] Its structural characterization data are as follows:

[0270] 1 H NMR (400MHz, DMSO): δ8.41(s,1H),7.70(s,1H),4.21(tt,J=5.8,2.8Hz,1H),3.88(s,3H),0.92(m,2H),0.80(m,2H).

[0271] Step 2: Preparation of 5-cyclopropyloxy-2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid methyl ester (7-11-2)

[0272] Methyl 4-bromo-5-cyclopropyloxy-2-nitrobenzoate (750 mg, 2.37 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (1.5 g, 5.93 mmol), bistriphenylphosphine palladium dichloride (499.6 mg, 0.71 mmol), and potassium acetate (700 mg, 7.12 mmol) were added to a single-necked flask and added to 1,4-dioxane (15 mL). The atmosphere was replaced with nitrogen three times, the temperature was raised to 101 ° C. and the reaction was carried out for 16 hours. The mixture was cooled to room temperature, filtered through celite, and concentrated to obtain a crude product (1.6 g, 4.41 mmol), which was used directly in the next step without purification.

[0273] Step 3: Preparation of 5-cyclopropyloxy-4-hydroxy-2-nitrobenzoic acid methyl ester (7-11-3)

[0274] Methyl 5-cyclopropyloxy-2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (1.6 g, 4.41 mmol) was dissolved in tetrahydrofuran (40 mL). Glacial acetic acid (5 mL) was added at 0°C, and hydrogen peroxide (10 mL) was slowly added dropwise. The reaction mixture was reacted at room temperature for 3 hours. Saturated sodium sulfite solution (50 mL) was added to the reaction solution and stirred for 1 hour. The mixture was concentrated under reduced pressure until no tetrahydrofuran remained. 30 mL of ethyl acetate was added for extraction. After separation, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (ethyl acetate / petroleum ether = 0% to 70%) and concentrated to obtain the title compound (1.05 g, 4.15 mmol).

[0275] Its structural characterization data are as follows:

[0276] ESI-MS (m / z): 253.2 [MH] -

[0277] Step 4: Preparation of methyl 4-(benzyloxy)-5-cyclopropyloxy-2-nitrobenzoate (7-11-4)

[0278] Methyl 5-cyclopropyloxy-4-hydroxy-2-nitrobenzoate (1.05 g, 4.15 mmol) was dissolved in DMF (5 mL), and potassium carbonate (1.2 g, 8.6 mmol) and benzyl bromide (1.49 g, 8.6 mmol) were added. The mixture was reacted at room temperature for 15 hours. After adding 30 mL of water, 50 mL of ethyl acetate was added for extraction. After separation, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product, which was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0% to 50%) to obtain the title compound (1.09 g, 3.17 mmol).

[0279] Step 5: Preparation of 4-(benzyloxy)-5-cyclopropyloxy-2-nitrobenzoic acid (7-11-5)

[0280] Methyl 4-(benzyloxy)-5-cyclopropyloxy-2-nitrobenzoate (1.09 g, 3.17 mmol) was dissolved in tetrahydrofuran (8 mL) and water (2 mL), and lithium hydroxide (152.1 mg, 6.35 mmol) was added. The mixture was reacted at room temperature for 3 hours. The pH of the system was adjusted to 5 with dilute hydrochloric acid, and a solid precipitated. The solid was extracted with 60 mL of ethyl acetate. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (743 mg, 2.26 mmol), which was used directly in the next step without purification.

[0281] Step 6: Preparation of (S)-5-(4-(benzyloxy)-5-cyclopropyloxy-2-nitrobenzoyl)-5-azaspiro[2.4]heptane-6-carboxylic acid methyl ester (7-11-6)

[0282] 4-(Benzyloxy)-5-cyclopropyloxy-2-nitrobenzoic acid (743 mg, 2.26 mmol) and (S)-5-azaspiro[2.4]heptane-6-carboxylic acid methyl ester (700 mg, 4.51 mmol) were dissolved in DMF (5 mL), and HATU (1.71 g, 4.51 mmol) and DIPEA (875 mg, 6.77 mmol) were added. The mixture was reacted at room temperature for 2 hours. After adding 30 mL of water, 50 mL of ethyl acetate was added for extraction. After separation, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was purified on a silica gel column (ethyl acetate / petroleum ether = 0% to 50%) and concentrated to obtain the title compound (910 mg, 1.95 mmol).

[0283] Its structural characterization data are as follows:

[0284] ESI-MS (m / z): 467.1 [M+H] +

[0285] Step 7: Preparation of (S)-8-(benzyloxy)-7-cyclopropyloxy-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5,11(10H)-dione (7-11-7)

[0286] Methyl (S)-5-(4-(Benzyloxy)-5-cyclopropyloxy-2-nitrobenzoyl)-5-azaspiro[2.4]heptane-6-carboxylate (910 mg, 1.95 mmol) was dissolved in methanol (30 mL), and a 10% aqueous solution of ammonium chloride (6 mL) and zinc powder (1.28 g, 19.51 mmol) were added. The temperature was raised to 40°C for reaction for 1 hour, then the temperature was raised to 80°C for reaction overnight, and the crude product was obtained by concentration. 35 mL of ethyl acetate was added to dissolve the product, and the product was washed three times with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was purified by reverse phase column chromatography (acetonitrile / 1% aqueous formic acid solution = 0% to 70%) and lyophilized to obtain the title compound (467 mg, 1.15 mmol).

[0287] Its structural characterization data are as follows:

[0288] ESI-MS (m / z): 405.2 [M+H] +

[0289] Step 8: Preparation of (S)-8-(benzyloxy)-7-cyclopropyloxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one (7-11-8)

[0290] (S)-8-(Benzyloxy)-7-cyclopropyloxy-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5,11(10H)-dione (467 mg, 1.15 mmol) was dissolved in dry tetrahydrofuran (20 mL). Sodium borohydride (128 mg, 3.46 mmol) was added, and trifluoroacetic acid (264 mg, 2.31 mmol) was slowly added dropwise, resulting in the generation of a large amount of gas. After the gas dissipated, the temperature was slowly raised to 70°C and refluxed for 16 hours. The reaction mixture was cooled to room temperature, quenched by the addition of methanol, and concentrated under reduced pressure. The product was purified by reverse-phase column chromatography (acetonitrile / 1% aqueous formic acid = 0% to 80%) and freeze-dried to afford the title compound (380 mg, 0.97 mmol).

[0291] Its structural characterization data are as follows:

[0292] ESI-MS (m / z): 363.3 [M+H] +

[0293] Step 9: Preparation of (S)-7-cyclopropyloxy-8-hydroxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one (7-11-9)

[0294] Dissolve (S)-8-(Benzyloxy)-7-cyclopropyloxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one (380 mg, 0.97 mmol) in dichloromethane (1.5 mL) and cool to 0°C. Then, add methanesulfonic acid (0.6 mL) dropwise. Incubate for 1 hour. Add water (10 mL) to the reaction mixture, then adjust the pH to 8 with saturated sodium bicarbonate solution. Extract three times with dichloromethane (8 mL x 3). The combined organic phases are washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude title compound (118 mg, 0.31 mmol), which is used directly in the next step without purification.

[0295] Its structural characterization data are as follows:

[0296] ESI-MS (m / z): 301.1 [M+H] +

[0297] Step 10: Preparation of tert-butyl (4-((S)-8-((5-(((S)-7-cyclopropyloxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-11-10)

[0298] Tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxy-50,11,11-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (141 mg, 0.23 mmol) and (S)-7-cyclopropyloxy-8-hydroxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (118 mg, 0.31 mmol) were dissolved in DMF (2 mL). Potassium carbonate (49 mg, 0.47 mmol) was added and the reaction was stirred for 16 hours. The reaction was quenched by adding water, extracted with ethyl acetate three times (20 mL x 3), and the organic phases were combined, washed with saturated brine, dried, and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) afforded the title compound (81 mg, 0.09 mmol).

[0299] Its structural characterization data are as follows:

[0300] ESI-MS (m / z): 806.3 [M+H] +

[0301] Step 11: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((S)-7-cyclopropyloxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-11)

[0302] Tert-butyl (4-((S)-8-((5-(((S)-7-cyclopropyloxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (81 mg, 0.09 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (0.6 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. After concentration under reduced pressure, purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%) and lyophilization, the title compound (3.5 mg, 0.005 mmol) was obtained.

[0303] Its structural characterization data are as follows:

[0304] ESI-MS (m / z): 706.3 [M+H]+

[0305] 1 H NMR (400MHz, DMSO): δ7.58(s,1H),7.27(d,J=11.8Hz,2H),7.11(d,J=8.4Hz,2H),6.57-6.51( m,2H),6.29(d,J=9.0Hz,2H),5.19(s,2H),4.18-4.07(m,1H),4.01-3.88(m,4H),3.86-3.80( m,1H),3.67(s,3H),3.57-3.51(m,4H),3.27-3.10(m,4H),2.73-2.65(m,1H),2.02-1.93(m,1 H),1.88-1.71(m,5H),1.61-1.48(m,2H),1.24(s,2H),0.88-0.75(m,1H),0.69-0.48(m,6H).

[0306] The preparation method is as follows:

[0307] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0308] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0309] Preparation Example 6: Preparation of (S)-8-((5-(((S)-2-(4-aminophenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one-11,11-d2(7-21)

[0310] Step 1: Preparation of (S)-8-(benzyloxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one-11,11-d2(7-21-1)

[0311] Dissolve (S)-8-(Benzyloxy)-7-methoxy-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diaza-2,1'-cyclopropane]-5,11(10H)-dione (1.00 g, 2.64 mmol) in tetrahydrofuran (3.00 mL). After cooling to 0°C, deuterated borane in tetrahydrofuran (0.75 M, 7.05 mL) was added dropwise under nitrogen. The mixture was returned to 25°C and stirred for 2 hours. Methanol (100 mL) was added to the reaction system, followed by stirring for another half hour. The mixture was then concentrated to afford the crude title compound (1.00 g), which was used directly in the next step without purification.

[0312] Its structural characterization data are as follows:

[0313] ESI-MS (m / z): 367.2 [M+H] +

[0314] Step 2: Preparation of (S)-8-hydroxy-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one-11,11-d2(7-21-2)

[0315] (S)-8-(Benzyloxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one-11,11-d2 (1.00 g, 2.73 mmol) was dissolved in methanol (10.0 mL). The atmosphere was purged with nitrogen three times, and then Pd / C (0.20 g, 187 μmol, 10% purity) was added. Hydrogen was then introduced and the reaction continued for 2 hours. The reaction solution was filtered through a silica gel pad and concentrated directly to obtain the crude product. After purification on a silica gel column (dichloromethane / methanol = 100 / 1 to 97 / 3), the product was concentrated again to obtain the title compound (500 mg, 1.77 mmol).

[0316] Its structural characterization data are as follows:

[0317] ESI-MS (m / z): 277.1 [M+H] +

[0318] 1H NMR: (400MHz, CD3OD) δ7.36(s,1H),6.52(s,1H),3.95-4.04(m,1H),3.86(s,3H),3.63(d,J=11.6Hz,1H),3.49(d ,J=12.0Hz,1H),2.26(dd,J=12.8,8.0Hz,1H),1.68(dd,J=12.8,4.4Hz,1H),0.69-0.79(m,2H),0.62-0.69(m,2H)

[0319] Step 3: Preparation of tert-butyl (4-((S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl-11,11-d2)oxy)pentyl)oxy)-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-2-yl)phenyl)carbamate (7-21-3)

[0320] Tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (21.23 mg, 36.19 μmol) and (S)-8-hydroxy-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one-11,11-d2 (10 mg, 36.19 μmol) were dissolved in DMF (1 mL). Potassium carbonate (14.98 mg, 108.57 μmol) was added and the mixture was stirred for 16 hours. The reaction was quenched by adding water, extracted with ethyl acetate three times (10 mL x 3), and the organic phases were combined, washed with saturated brine, dried, and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) afforded the title compound (20 mg, 25.58 μmol).

[0321] Its structural characterization data are as follows:

[0322] ESI-MS (m / z): 794.4 [M+H] +

[0323] Step 4: Preparation of (S)-8-((5-(((S)-2-(4-aminophenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one-11,11-d2(7-21)

[0324] Tert-butyl (4-((S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-8-yl-11,11-d2)oxy)pentyl)oxy)-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (20 mg, 25.58 μmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (0.5 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. After concentration under reduced pressure, purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%) and lyophilization, the title compound (4.7 mg, 6.82 μmol) was obtained.

[0325] Its structural characterization data are as follows:

[0326] ESI-MS (m / z): 682.4 [M+H] +

[0327] 1 H NMR (400MHz, DMSO) δ7.28(d,J=13.4Hz,2H),7.27(s,1H),7.15(d,J=8.5Hz,2H),6. 59(d,J=8.6Hz,2H),6.30(d,J=10.2Hz,2H),3.95(dd,J=12.0,6.1Hz,4H),3.66(d,J =7.5Hz,6H),3.59-3.51(m,2H),3.37-3.15(m,4H),2.76-2.67(m,2H),2.03-1.92( m,2H),1.85-1.75(m,4H),1.62-1.52(m,2H),0.68-0.58(m,2H),0.56-0.48(m,2H).

[0328] The preparation method is as follows:

[0329] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0330] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0331] Preparation Example 7: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-22)

[0332] Step 1: Preparation of (4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-L-proline methyl ester (7-22-2)

[0333] 4-(Benzyloxy)-5-methoxy-2-nitrobenzoic acid (3.00 g, 9.89 mmol) and L-proline methyl ester hydrochloride (1.97 g, 11.87 mmol) were dissolved in DMF (30 mL), and HATU (5.64 g, 14.84 mmol) and DIPEA (3.84 g, 29.68 mmol) were added in sequence. The mixture was reacted at 25 ° C for 1 hour. The reaction solution was poured into water (100 ml), and the product was extracted with EA / PE = 1 / 1 (200 ml), washed with brine (30 ml), and concentrated to give the title compound (4.10 g, 9.89 mmol), which was used directly in the next step without purification.

[0334] Its structural characterization data are as follows:

[0335] MS m / z(ESI):415.1[M+H] +

[0336] Step 2: Preparation of (S)-8-hydroxy-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-22-3)

[0337] The compound (4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-L-proline methyl ester (3.80 g, 8.98 mmol) was dissolved in methanol (40 mL) and tetrahydrofuran (40 mL), and 10% palladium on carbon (380 mg) and HOAc (1.08 g, 17.95 mmol) were added. The air was removed under vacuum, and hydrogen was introduced. The reaction was carried out at 25°C for 16 hours. The palladium on carbon was filtered off, and the filtrate was concentrated to give the title compound (2.35 g, 8.98 mmol), which was used directly in the next step without purification.

[0338] Its structural characterization data are as follows:

[0339] MS m / z(ESI):263.1[M+H] +

[0340] Step 3: Preparation of (S)-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-22-4)

[0341] Compound (S)-8-hydroxy-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (100.0 mg, 381.30 μmol) was dissolved in THF (3 mL), and borane tetrahydrofuran solution (4 M, 476.62 μL) was added. The reaction was stirred at 25°C for 1 hour, and the reaction solution was poured into methanol (3 ml) for destruction. The pH was adjusted to 5-6 with 20% formic acid aqueous solution, and the crude product was concentrated to obtain the crude product. The crude product was purified by C18 reverse phase column (H2O:ACN=0-30%, 0.05% FA) and lyophilized to obtain the title compound (95.0 mg, 306.11 μmol).

[0342] Its structural characterization data are as follows:

[0343] MS m / z(ESI):249.1[M+H] +

[0344] Step 4: Preparation of tert-butyl (4-((S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-22-5)

[0345] (S)-8-((5-bromopentyl)oxy)-2-(4-((tert-butoxycarbonyl)amino)phenyl)-7-methoxy-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (20 mg, 34.10 μmol), compound (S)-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4 Diazepin-5-one (12.70 mg, 51.15 μmol, FR) was dissolved in DMF (1 mL), and Cs2CO3 (33.33 mg, 102.30 μmol) was added. The temperature was raised to 40°C and the reaction was allowed to react for 16 hours. EA (20 ml) and H2O (6 ml) were added to the system and stirred for 5 minutes. The EA phase was separated, washed with brine, and concentrated to give the crude product, which was purified by thin layer chromatography (MeOH / DCM = 10%) to give the title compound (15.0 mg, 19.90 μmol).

[0346] Its structural characterization data are as follows:

[0347] MS m / z(ESI):754.4[M+H] +

[0348] Step 5: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-22)

[0349] Compound tert-butyl (4-((S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (15.0 mg, 19.90 μmol) was dissolved in DCM (0.5 mL) and trifluoroacetic acid (226.87 mg, 1.99 mmol) was added. The reaction was carried out at 25°C for 1 hour, and the crude product was concentrated to give the title compound (7.10 mg, 10.53 μmol). The crude product was purified by preparative HPLC and freeze-dried to give the title compound (7.10 mg, 10.53 μmol).

[0350] The preparation method is as follows:

[0351] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0352] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0353] Its structural characterization data are as follows:

[0354] MS m / z(ESI):654.4[M+H] +

[0355] 1 H NMR (400MHz, MeOD): δ7.38 (dd, J=8.2, 2.3Hz, 3H), 6.91 (d, J=8.9Hz, 2H), 6.27 (d, J=9.3Hz, 2H), 4.24 (s, 1H),4.00(dd,J=9.9,6.3Hz,4H),3.80(s,3H),3.77(s,3H),3.76(s,3H),3.64(s,1H),3.61(s,1H),3.56 (d,J=11.1Hz,2H),3.50-3.46(m,2H),3.41(s,1H),3.40-3.33(m,3H),2.80(d,J=13.5Hz,1H),2.08(d,J =5.0Hz,1H),1.95-1.86(m,5H),1.69(d,J=6.7Hz,2H),0.71-0.68(m,1H),0.61(dd,J=12.4,6.0Hz,3H).

[0356] Preparation Example 8: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2R,11aS)-2-hydroxy-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-23)

[0357] Step 1: Preparation of methyl (2S,4R)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-hydroxypyrrolidine-2-carboxylate (7-23-2)

[0358] Dissolve 4-benzyloxy-5-methoxy-2-nitro-benzoic acid (10 g, 32.97 mmol), (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid methyl ester hydrochloride (5.27 g, 36.27 mmol), and HATU (18.81 g, 49.46 mmol) in DMF (50 mL). Add DIPEA (12.78 g, 98.92 mmol) dropwise and stir for 2 hours. Add water and EA, stir, and separate. The aqueous phase is extracted twice with EA. The combined organic phases are washed twice with saturated brine, dried, and concentrated. Purify on a silica gel column (eluent: 30-65% ethyl acetate / petroleum ether) and reconcentrate to obtain the title compound (14 g, 32.53 mmol).

[0359] Step 2: Preparation of (2R,11aS)-8-(benzyloxy)-2-hydroxy-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-23-3)

[0360] Dissolve methyl (2S,4R)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-hydroxypyrrolidine-2-carboxylate (14 g, 32.53 mmol) in methanol (700 mL). Add saturated aqueous ammonium chloride (70 mL) and zinc powder (21.27 g, 325.27 mmol) in portions. Heat to 80°C and reflux for 16 hours. Cool to room temperature, filter, and concentrate the filtrate under reduced pressure. Purify on a silica gel column (eluent: ethyl acetate) to obtain the title compound (5.3 g, 14.39 mmol).

[0361] Its structural characterization data are as follows:

[0362] MS m / z(ESI):368.5[M+H] +

[0363] Step 3: Preparation of (2R,11aS)-8-(benzyloxy)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-23-4)

[0364] Dissolve (2R,11aS)-8-(benzyloxy)-2-hydroxy-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (1 g, 2.71 mmol) in DMF (25 mL). Add imidazole (1.85 g, 27.15 mmol) and TBSCl (1.12 g, 13.57 mmol) and allow to react for 16 hours. Add water and ethyl acetate, stir, and allow to stand for separation. The organic phase is washed three times with saturated brine and concentrated under reduced pressure. Purify on a silica gel column (eluent: 50% ethyl acetate / petroleum ether) and concentrate again to obtain the title compound (1.02 g, 2.11 mmol).

[0365] Step 4: Preparation of (2R,11aS)-8-(benzyloxy)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-23-5)

[0366] Dissolve (2R,11aS)-8-(benzyloxy)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (500 mg, 1.04 mmol) in tetrahydrofuran (5 mL) and cool to 0°C. Then, add tetrahydrofuran borane complex (10.3 mL) dropwise. After addition, warm to room temperature and react for 3 hours. Methanol (10 ml) is added dropwise to the reaction solution, which is then concentrated to obtain the crude title compound. Purify on a silica gel column (ethyl acetate / petroleum ether = 0% to 70%) and concentrate to obtain the title compound (120 mg, 0.26 mmol).

[0367] Its structural characterization data are as follows:

[0368] ESI-MS (m / z): 453.3 [M+H] +

[0369] Step 5: Preparation of (2R,11aS)-2-((tert-butyldimethylsilyl)oxy)-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-23-6)

[0370] Dissolve (2R,11aS)-8-(benzyloxy)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (120 mg, 0.26 mmol) in tetrahydrofuran (3 mL). Add 20% palladium on carbon (12 mg). Replace the atmosphere with hydrogen three times, then heat to 45°C and stir for 3 hours. Filter through celite and concentrate to obtain the crude title compound (90 mg, 0.23 mmol), which is used directly in the next step without purification.

[0371] Its structural characterization data are as follows:

[0372] ESI-MS (m / z): 379.2 [M+H] +

[0373] Step 6: Preparation of tert-butyl (4-((S)-8-((5-(((2R,11aS)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-2,3,5),10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-23-7)

[0374] Tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxy-50,11,11-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (135 mg, 0.23 mmol) and (2R,11aS)-2-((tert-butyldimethylsilyl)oxy)-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (90 mg, 0.23 mmol) were dissolved in DMF (2 mL). Potassium carbonate (64 mg, 0.47 mmol) was added and the mixture was stirred for 16 hours. The reaction was quenched by adding water, extracted with ethyl acetate three times (20 mL x 3), and the organic phases were combined, washed with saturated brine, dried, concentrated, and purified on a silica gel column (MeOH / DCM = 0% to 5%) and concentrated again to obtain the title compound (82 mg, 0.092 mmol).

[0375] Its structural characterization data are as follows:

[0376] ESI-MS (m / z): 884.3 [M+H] +

[0377] Step 7: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2R,11aS)-2-hydroxy-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-23)

[0378] Tert-butyl (4-((S)-8-((5-(((2R,11aS)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-2,3,5),10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (82 mg, 0.092 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (3 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 5 hours. The residue was concentrated under reduced pressure, purified by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%), and lyophilized to obtain the title compound (8.4 mg, 0.012 mmol).

[0379] Its structural characterization data are as follows:

[0380] ESI-MS (m / z): 670.3 [M+H] +

[0381] 1H NMR (400MHz, DMSO): δ7.37(s,1H),7.29(s,1H),7.26(s,1H),7.11(d,J=8.4Hz,2H),6.53(d,J=8.4Hz, 2H),6.31(s,1H),6.26(s,1H),5.28-5.10(m,2H),5.08-4.79(m,1H),4.21-4.11(m,2H),3.97-3.88(m ,4H),3.89-3.76(m,1H),3.66(s,3H),3.64(s,3H),3.55-3.47(m,4H),3.25-3.20(m,2H),3.01-2.93( m,1H),2.68-2.65(m,1H),2.12-2.04(m,1H),1.85-1.76(m,4H),1.74-1.66(m,1H),1.61-1.52(m,2H).

[0382] The preparation method is as follows:

[0383] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0384] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0385] Preparation Example 9: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2S,11aS)-2-hydroxy-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-24)

[0386] Step 1: Preparation of methyl (2S,4S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-hydroxypyrrolidine-2-carboxylate (7-24-2)

[0387] 4-(Benzyloxy)-5-methoxy-2-nitrobenzoic acid (1.0 g, 3.3 mmol) and methyl (2S,4S)-4-hydroxypyrrolidine-2-carboxylate (503 mg, 3.46 mmol) were dissolved in DMF (10 mL), and HATU (2.51 g, 6.51 mmol) and DIPEA (1.28 g, 9.9 mmol) were added. The mixture was reacted at room temperature for 2 hours. After adding 50 mL of water, 70 mL of ethyl acetate was added for extraction. After separation, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was purified on a silica gel column (ethyl acetate / petroleum ether = 0% to 50%) and concentrated to obtain the title compound (1.36 g, 3.16 mmol).

[0388] Its structural characterization data are as follows:

[0389] ESI-MS (m / z): 431.1 [M+H] +

[0390] Step 2: Preparation of (2S,11aS)-8-(benzyloxy)-2-hydroxy-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-24-3)

[0391] Methyl (2S,4S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-hydroxypyrrolidine-2-carboxylate (1.36 g, 3.16 mmol) was dissolved in methanol (52 mL), and a 10% aqueous solution of ammonium chloride (10 mL) and zinc powder (2.43 g, 37.2 mmol) were added. The temperature was raised to 40°C for reaction for 1 hour, then the temperature was raised to 80°C for reaction overnight, and the crude product was obtained by concentration. 50 mL of ethyl acetate was added to dissolve the product, and the product was washed three times with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The product was purified by reverse phase column chromatography (acetonitrile / 1% aqueous formic acid solution = 0% to 70%) and lyophilized to obtain the title compound (1.3 g, 3.53 mmol).

[0392] Its structural characterization data are as follows:

[0393] ESI-MS (m / z): 369.2 [M+H] +

[0394] Step 3: Preparation of (2S,11aS)-8-(benzyloxy)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-24-4)

[0395] (2S,11aS)-8-(Benzyloxy)-2-hydroxy-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (1.3 g, 3.53 mmol) was dissolved in DMF (15 mL), and tert-butyldimethylsilyl chloride (2.66 g, 17.64 mmol) and imidazole (2.4 g, 35.3 mmol) were added. After reacting at room temperature for 16 hours, 60 mL of water was added, and then 80 mL of ethyl acetate was added for extraction. After separation, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was purified on a silica gel column (ethyl acetate / petroleum ether = 0% to 50%) to obtain the title compound (1.38 g, 2.86 mmol).

[0396] Its structural characterization data are as follows:

[0397] ESI-MS (m / z): 483.2 [M+H] +

[0398] Step 4: Preparation of (2S,11aS)-8-(benzyloxy)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5-one(7-24-5)

[0399] Dissolve (2S,11aS)-8-(benzyloxy)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (500 mg, 1.04 mmol) in tetrahydrofuran (5 mL) and cool to 0°C. Then, add tetrahydrofuran borane complex (10.3 mL) dropwise. After addition, warm to room temperature and react for 3 hours. Methanol (10 ml) is added dropwise to the reaction solution, which is then concentrated to obtain the crude title compound. Purify on a silica gel column (ethyl acetate / petroleum ether = 0% to 70%) and concentrate to obtain the title compound (60 mg, 0.13 mmol).

[0400] Its structural characterization data are as follows:

[0401] ESI-MS (m / z): 469.2 [M+H] +

[0402] Step 5: Preparation of (2S,11aS)-2-((tert-butyldimethylsilyl)oxy)-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-24-6)

[0403] Dissolve (2S,11aS)-8-(benzyloxy)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (60 mg, 0.13 mmol) in tetrahydrofuran (3 mL). Add 20% palladium on carbon (6 mg). Replace the atmosphere with hydrogen three times, then heat to 25°C and stir for 2 hours. Filter through celite and concentrate to obtain the crude title compound (46 mg, 0.12 mmol), which is used directly in the next step without purification.

[0404] Its structural characterization data are as follows:

[0405] ESI-MS (m / z): 379.2 [M+H] +

[0406] Step 6: Preparation of tert-butyl (4-((S)-8-((5-(((2S,11aS)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-24-6)

[0407] Tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxy-50,11,11-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (71 mg, 0.12 mmol) and (2S,11aS)-2-((tert-butyldimethylsilyl)oxy)-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (46 mg, 0.12 mmol) were dissolved in DMF (2 mL). Potassium carbonate (33 mg, 0.24 mmol) was added and the mixture was stirred for 16 hours. The reaction was quenched by adding water, and the organic phases were combined after extraction with ethyl acetate three times (20 mL x 3), washed with saturated brine, and then dried and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) gave the title compound (39 mg, 0.04 mmol).

[0408] Its structural characterization data are as follows:

[0409] ESI-MS (m / z): 884.3 [M+H] +

[0410] Step 7: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2S,11aS)-2-hydroxy-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-24)

[0411] Tert-butyl (4-((S)-8-((5-(((2S,11aS)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (39 mg, 0.04 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (2 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 5 hours. The residue was concentrated under reduced pressure, purified by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%), and lyophilized to obtain the title compound (7 mg, 0.01 mmol).

[0412] Its structural characterization data are as follows:

[0413] ESI-MS (m / z): 670.3 [M+H]+

[0414] 1 H NMR (400MHz, DMSO): δ7.28(d,J=4.0Hz,2H),7.26(s,1H),7.12(d,J=8.4Hz,2H),6.54(d,J=8.4Hz,2H ),6.30(s,1H),6.26(s,1H),5.37-5.14(m,1H),5.10-5.03(m,1H),4.24-4.06(m,2H),3.96-3.84(m,2 H),3.66(s,3H),3.63(s,3H),3.61-3.56(m,1H),3.55-3.48(m,1H),3.47-3.40(m,3H),3.26-3.21(m ,3H),2.68-2.64(m,1H),2.34-2.30(m,1H),1.85-1.76(m,4H),1.71-1.63(m,1H),1.62-1.51(m,2H).

[0415] The preparation method is as follows:

[0416] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0417] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0418] Preparation Example 10: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((2S,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-25)

[0419] Step 1: Preparation of methyl (2S, 4R)-4-methoxypyrrolidine-2-carboxylate (7-25-2)

[0420] Dissolve 1-(tert-butyl)-2-methyl(2S,4R)-4-methoxypyrrolidine-1,2-dicarboxylic acid (1.0 g, 3.53 mmol, FR) in methanol (10 mL), add HCl (4M in EA) (5 mL), and react at room temperature for 1 hour. Concentrate to obtain the crude product, which is used directly in the next step without purification (800.6 mg, 5.01 mmol).

[0421] Its structural characterization data are as follows:

[0422] ESI-MS (m / z): 160.1 [M+H] +

[0423] Step 2: Preparation of (2R,4S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-methoxypyrrolidine-2-carboxylic acid methyl ester (7-25-3)

[0424] Methyl (2S,4R)-4-methoxypyrrolidine-2-carboxylate (800.6 mg, 5.01 mmol) and 4-(benzyloxy)-5-methoxy-2-nitrobenzoic acid (1.18 g, 3.88 mmol, FR) were dissolved in DMF (10 mL). HATU (1.48 g, 3.88 mmol) and DIPEA (1.36 g, 10.59 mmol) were added and reacted at room temperature for 1 hour. The reaction solution was added to ethyl acetate (100 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (ethyl acetate / petroleum ether = 20% to 60%) and concentrated again to obtain the title compound (1.56 g, 3.51 mmol).

[0425] Its structural characterization data are as follows:

[0426] ESI-MS (m / z): 445.2 [M+H] +

[0427] Step 3: Preparation of (2R,11aS)-8-(benzyloxy)-2,7-dimethoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-25-4)

[0428] Methyl (2S,4R)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-(trifluoromethyl)pyrrolidine-2-carboxylate (1.56 g, 3.1 mmol) was dissolved in methanol (30 mL). Zinc powder (2.4 g, 31.1 mmol) was added, followed by a saturated aqueous solution of ammonium chloride (40 mL). The mixture was heated to 85°C and refluxed for 16 hours. The reaction mixture was added to ethyl acetate (150 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude title compound (1.13 g, 2.95 mmol), which was used directly in the next step without purification.

[0429] Its structural characterization data are as follows:

[0430] ESI-MS (m / z): 383.2 [M+H] +

[0431] Step 4: Preparation of (2R,11aS)-8-(benzyloxy)-2,7-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-25-5)

[0432] Dissolve (2R,11aS)-8-(benzyloxy)-2,7-dimethoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (0.5 g, 1.3 mmol, FR) in tetrahydrofuran (5 mL) and cool to 0°C. Then, add tetrahydrofuran borane complex (10.3 mL) dropwise. After addition, warm to room temperature and react for 3 hours. Add methanol (10 mL) dropwise to the reaction solution, then concentrate to obtain the crude title compound. Purify on a silica gel column (ethyl acetate / petroleum ether = 0% to 70%) and concentrate to obtain the title compound (80 mg, 0.22 mmol).

[0433] Its structural characterization data are as follows:

[0434] ESI-MS (m / z): 369.1 [M+H] +

[0435] Step 5: Preparation of (2R,11aS)-8-hydroxy-2,7-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-25-6)

[0436] Dissolve (2R,11aS)-8-(benzyloxy)-2,7-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (80 mg, 0.22 mmol) in tetrahydrofuran (3 mL), add palladium on carbon (8 mg), and react under a hydrogen atmosphere for 1 hour. After completion, filter the reaction, dry the filtrate over anhydrous sodium sulfate, filter, and concentrate to obtain the crude title compound (49 mg, 0.17 mmol), which was used directly in the next step without purification.

[0437] Its structural characterization data are as follows:

[0438] ESI-MS (m / z): 279.2 [M+H] +

[0439] Step 6: Preparation of tert-butyl (4-((S)-8-((5-(((2R,11aS)-2,7-dimethoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H)-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-26-7)

[0440] Tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (30.8 mg, 0.053 mmol) and (2R,11aS)-8-hydroxy-2,7-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (15 mg, 0.05 mmol) were dissolved in DMF (1.5 mL). Potassium carbonate (13.8 mg, 0.1 mmol) was added and the mixture was stirred for 16 hours. The reaction was quenched by adding water, and the organic phases were combined after extraction with ethyl acetate three times (10 mL x 3), washed with saturated brine, and then dried and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) was followed by further concentration to afford the title compound (19.6 mg, 0.25 mmol).

[0441] Its structural characterization data are as follows:

[0442] ESI-MS (m / z): 784.4 [M+H] +

[0443] Step 7: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((2S,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-25)

[0444] Tert-butyl (4-((S)-8-((5-(((2R,11aS)-2,7-dimethoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H)-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (19.6 mg, 0.25 mmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (1 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. After concentration under reduced pressure, purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%) and lyophilization, the title compound (9.2 mg, 13.16 μmol) was obtained.

[0445] Its structural characterization data are as follows:

[0446] ESI-MS (m / z): 684.4 [M+H] +

[0447] 1 H NMR (400MHz, DMSO): δ7.37(s,1H),7.29(s,1H),7.26(s,1H),7.11(d,J=8.4Hz,2H),6.53(d,J =8.4Hz,2H),6.30(s,1H),6.26(s,1H),5.19(s,2H),4.19-4.08(m,1H),3.96-3.84(m,5H),3.7 4-3.68(m,2H),3.66(s,3H),3.64(s,3H),3.56-3.46(m,3H),3.29-3.22(m,2H),3.22(s,3H),3 .02-2.93(m,1H),2.73-2.66(m,1H),1.85-1.76(m,4H),1.75-1.66(m,1H),1.61-1.50(m,2H).

[0448] The preparation method is as follows:

[0449] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0450] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0451] Preparation Example 11: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2S,11aS)-2,7-dimethoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-26)

[0452] Step 1: Preparation of methyl (2S,4S)-4-(methoxy)pyrrolidine-2-carboxylate (7-26-2)

[0453] Dissolve 1-(tert-butyl)-2-methyl(2S,4S)-4-(methoxy)pyrrolidine-1,2-dicarboxylic acid (1.0 g, 4.08 mmol) in methanol (10 mL), add HCl (4M in EA) (5 mL), and react at room temperature for 1 hour. Concentrate to obtain the crude product, which is used directly in the next step without purification (797.66 mg, 4.08 mmol).

[0454] Its structural characterization data are as follows:

[0455] ESI-MS (m / z): 196.2 [M+H] +

[0456] Step 2: Preparation of (2S,4S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-(methoxy)pyrrolidine-2-carboxylic acid methyl ester (7-26-3)

[0457] Methyl (2S,4S)-4-(methyloxy)pyrrolidine-2-carboxylate (797.66 mg, 4.08 mmol) and 4-(benzyloxy)-5-methoxy-2-nitrobenzoic acid (1.24 g, 4.08 mmol) were dissolved in DMF (10 mL). HATU (1.86 g, 4.89 mmol) and DIPEA (1.58 g, 12.23 mmol) were added and reacted at room temperature for 1 hour. The reaction solution was added to ethyl acetate (100 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (ethyl acetate / petroleum ether = 20% to 60%) and concentrated again to obtain the title compound (1.8 g, 4.05 mmol).

[0458] Its structural characterization data are as follows:

[0459] ESI-MS (m / z): 445.2 [M+H] +

[0460] Step 3: Preparation of (2S,11aS)-8-(benzyloxy)-7-methoxy-2-(methoxy)-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-26-4)

[0461] Methyl (2S,4R)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-(methoxy)pyrrolidine-2-carboxylate (1.8 g, 4.05 mmol) was dissolved in methanol (30 mL). Zinc powder (2.63 g, 40.50 mmol) was added, followed by a saturated aqueous solution of ammonium chloride (40 mL). The mixture was heated to 85°C and refluxed for 16 hours. The reaction mixture was added to ethyl acetate (150 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude title compound (1.5 g, 3.92 mmol), which was used directly in the next step without purification.

[0462] Its structural characterization data are as follows:

[0463] ESI-MS (m / z): 383.2 [M+H] +

[0464] Step 4: Preparation of (2S,11aS)-8-(benzyloxy)-2,7-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-26-5)

[0465] Dissolve (2S,11aS)-8-(benzyloxy)-2,7-dimethoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (0.8 g, 2.09 mmol) in tetrahydrofuran (10 mL), add sodium borohydride (158.13 mg, 41.8 mmol), then add trifluoroacetic acid (596 mg, 5.23 mmol) dropwise at room temperature until no more gas escapes, heat to 75°C and reflux for 5 hours, concentrate the reaction solution, add water (50 mL), and then use ethyl acetate (50 mL X 10 mL) to obtain the product. 2) Extract twice, wash the combined organic phases with saturated sodium chloride aqueous solution, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify by reverse-phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%), followed by lyophilization to obtain the title compound (640 mg, 1.73 mmol). Its structural characterization data are as follows:

[0466] ESI-MS (m / z): 369.1 [M+H] +

[0467] Step 5: Preparation of (2S,11aS)-8-hydroxy-2,7-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-26-6)

[0468] Dissolve (2S,11aS)-8-(benzyloxy)-2,7-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (640 mg, 1.73 mmol) in methanol (10 mL). Add palladium on carbon (30 mg) and react under a hydrogen atmosphere for 1 hour. After completion, filter the reaction, dry the filtrate over anhydrous sodium sulfate, filter, and concentrate to obtain the crude title compound (434 mg, 1.56 mmol), which is used directly in the next step without purification.

[0469] Its structural characterization data are as follows:

[0470] ESI-MS (m / z): 279.2 [M+H] +

[0471] Step 6: Preparation of tert-butyl (4-((S)-8-((5-(((2S,11aS)-2,7-dimethoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-26-7)

[0472] Tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (21.07 mg, 35.93 μmol) and (2S,11aS)-8-hydroxy-2,7-dimethoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (10 mg, 35.93 μmol) were dissolved in DMF (1 mL). Potassium carbonate (14.88 mg, 107.80 μmol) was added and the mixture was stirred for 16 hours. The reaction was quenched by adding water and extracted with ethyl acetate three times (10 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) was followed by further concentration to afford the title compound (20 mg, 25.51 μmol).

[0473] Its structural characterization data are as follows:

[0474] ESI-MS (m / z): 784.4 [M+H] +

[0475] Step 7: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2S,11aS)-2,7-dimethoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-26)

[0476] Tert-butyl (4-((S)-8-((5-(((2S,11aS)-2,7-dimethoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (20 mg, 25.51 μmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (0.5 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. After concentration under reduced pressure, purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%) and lyophilization, the title compound (13.51 mg, 19.56 μmol) was obtained.

[0477] Its structural characterization data are as follows:

[0478] ESI-MS (m / z): 684.4 [M+H] +

[0479] 1 H NMR (400MHz, DMSO): δ7.39 (s, 1H), 7.28 (d, J = 9.3Hz, 2H), 6.85-6.75 (m, 2H), 6.48- 6.63(m,2H),6.31(s,1H),6.29(s,1H)4.22-4.15(m,1H),3.97-3.86(m,4H),3.75-3 .65(m,2H),3.66(s,3H),3.64(s,3H),3.62-3.50(m,6H),3.24(s,3H),3.32-3.16( m,2H),2.76-2.67(m,1H),2.43-2.36(m,1H),1.87-1.73(m,4H),1.60-1.49(m,4H).

[0480] The preparation method is as follows:

[0481] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0482] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0483] Preparation Example 12: Preparation of (S)-8-((5-(((2R,11aS)-2-amino-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-2-(4-aminophenyl)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-27)

[0484] Step 1: Preparation of methyl (2S,4R)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-((tert-butoxycarbonyl)amino)pyrrolidine-2-carboxylate (7-27-2)

[0485] Methyl (2S,4R)-4-((tert-butoxycarbonyl)amino)pyrrolidine-2-carboxylate (1 g, 4.09 mmol, FR) and 4-(benzyloxy)-5-methoxy-2-nitrobenzoic acid (1.37 g, 4.50 mmol, FR) were dissolved in DMF (10 mL). HATU (1.71 g, 4.50 mmol) and DIPEA (1.59 g, 12.28 mmol) were added and reacted at room temperature for 1 hour. The reaction solution was added to ethyl acetate (50 mL) and saturated brine (50 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (ethyl acetate / petroleum ether = 20% to 60%) and concentrated again to obtain the title compound (2.1 g, 3.97 mmol).

[0486] Its structural characterization data are as follows:

[0487] ESI-MS (m / z): 530.2 [M+H] +

[0488] Step 2: Preparation of tert-butyl ((2R,11aS)-8-(benzyloxy)-7-methoxy-5,11-dioxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)carbamate (7-27-3)

[0489] Methyl (2S,4R)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-((tert-butoxycarbonyl)amino)pyrrolidine-2-carboxylate (2.1 g, 3.97 mmol, FR) was dissolved in methanol (50 mL). Zinc powder (2.58 g, 15.86 mmol) was added, followed by a saturated aqueous solution of ammonium chloride (20 mL). The temperature was raised to 85°C and refluxed for 16 hours. The reaction mixture was added to ethyl acetate (150 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude title compound (1.8 g, 3.85 mmol), which was used directly in the next step without purification.

[0490] Its structural characterization data are as follows:

[0491] ESI-MS (m / z): 468.3 [M+H]+

[0492] Step 3: Preparation of tert-butyl ((2R,11aS)-8-(benzyloxy)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)carbamate (7-27-4)

[0493] Dissolve tert-butyl ((2R,11aS)-8-(benzyloxy)-7-methoxy-5,11-dioxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)carbamate (1.8 g, 3.85 mmol, FR) in tetrahydrofuran (100 mL), add sodium borohydride (843.15 mg, 11.55 mmol), and then add trifluoroacetic acid (877.8 mg, 7.7 mmol) dropwise at room temperature until no more gas escapes. Heat to 75°C and reflux for 5 hours. The reaction solution was concentrated, water (50 mL) was added, and the mixture was extracted twice with ethyl acetate (50 mL x 2). The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude title compound (1.4 g, 3.09 mmol), which was used in the next step without purification.

[0494] Its structural characterization data are as follows:

[0495] ESI-MS (m / z): 454.2 [M+H] +

[0496] Step 4: Preparation of tert-butyl ((2R,11aS)-8-hydroxy-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)carbamate (7-27-5)

[0497] Dissolve tert-butyl ((2R,11aS)-8-(benzyloxy)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)carbamate (100 mg, 220.50 μmol, FR) in methanol (5 mL). Add palladium on carbon (5 mg) and react under a hydrogen atmosphere for 1 hour. After completion, filter the reaction, dry the filtrate over anhydrous sodium sulfate, filter, and concentrate to obtain the crude title compound (80.13 mg, 220.50 μmol), which is used directly in the next step without purification.

[0498] Its structural characterization data are as follows:

[0499] ESI-MS (m / z): 364.2 [M+H] +

[0500] Step 5: Preparation of tert-butyl (4-((S)-8-((5-(((2R,11aS)-2-((tert-butoxycarbonyl)amino)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-27-6)

[0501] Tert-butyl ((2R,11aS)-8-hydroxy-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)carbamate (10 mg, 27.52 μmol) and tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (16.14 mg, 27.52 μmol) were dissolved in DMF (1 mL). Potassium carbonate (11.39 mg, 82.55 μmol) was added and the mixture was stirred for 16 hours. The reaction was quenched by adding water, and the organic phases were combined after extraction with ethyl acetate three times (10 mL x 3), washed with saturated brine, and then dried and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) afforded the title compound (20 mg, 23.01 μmol).

[0502] Its structural characterization data are as follows:

[0503] ESI-MS (m / z): 869.8 [M+H] +

[0504] Step 6: Preparation of (S)-8-((5-(((2R,11aS)-2-amino-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-2-(4-aminophenyl)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-27)

[0505] Tert-butyl (4-((S)-8-((5-(((2R,11aS)-2-((tert-Butoxycarbonyl)amino)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (20 mg, 23.01 μmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (0.5 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. The residue was concentrated under reduced pressure, purified by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%), and lyophilized to obtain the title compound (5.41 mg, 0.045 mmol).

[0506] Its structural characterization data are as follows:

[0507] ESI-MS (m / z): 670.4 [M+H] +

[0508] 1 H NMR (400MHz, DMSO): δ7.36 (s, 1H), 7.29 (s, 1H), 7.25 (s, 1H), 7.11 (d, J = 8.5Hz, 2H), 6 .53(d,J=8.6Hz,2H),6.29(d,J=9.3Hz,2H),4.17-4.09(m,1H),3.96-3.90(m,4H),3.6 5(d,J=7.1Hz,6H),3.51(d,J=4.9Hz,2H),3.30-3.20(m,4H),2.99(dd,J=12.9,8.8Hz ,2H),2.23-2.14(m,2H),1.96-1.87(m,2H),1.84-1.76(m,4H),1.56(d,J=6.4Hz,2H).

[0509] The preparation method is as follows:

[0510] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0511] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0512] Preparation Example 13: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((S)-2,2-difluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-29)

[0513] Step 1: Preparation of (S)-4,4-difluoropyrrolidine-2-carboxylic acid methyl ester (7-29-2)

[0514] Dissolve (S)-1-(tert-Butoxycarbonyl)-4,4-difluoropyrrolidine-2-carboxylic acid (0.6 g, 2.39 mmol) in MeOH (3 mL), add 4M HCl / dioxane solution (4 mL), and stir at room temperature for 20 hours. After completion, the reaction mixture was concentrated under reduced pressure to obtain the crude hydrochloride salt of the title compound (460 mg, 2.28 mmol), which was used directly in the next step without purification.

[0515] Its structural characterization data are as follows:

[0516] ESI-MS (m / z): 166.1 [M+H] +

[0517] Step 2: Preparation of (S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4,4-difluoropyrrolidine-2-carboxylic acid methyl ester (7-29-3)

[0518] Dissolve 4-(Benzyloxy)-5-methoxy-2-nitrobenzoic acid (743.76 mg, 2.45 mmol) in DMF (5 mL), add HATU (1.03 g, 2.70 mmol) and DIPEA (316.96 mg, 2.45 mmol), and stir at room temperature for 10 minutes. In another reaction flask, dissolve (S)-methyl 4,4-difluoropyrrolidine-2-carboxylate hydrochloride (450 mg, 2.45 mmol) in DMF (5 mL), and add DIPEA (316.96 mg, 2.45 mmol) to make the solution alkaline. Combine the two reaction solutions and stir at room temperature for 2 hours. After completion, add 30 mL of water to the reaction solution, extract with 15 mL of ethyl acetate x 3 times, and wash the organic phase with 15 mL of saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain the crude title compound (1.4 g, 2.18 mmol), which is used directly in the next step without purification.

[0519] Its structural characterization data are as follows:

[0520] ESI-MS (m / z): 468.2 [M+H] +

[0521] Step 3: Preparation of (S)-8-(benzyloxy)-2,2-difluoro-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-29-4)

[0522] Methanol (12 mL) and saturated aqueous ammonium chloride (3 mL) were added to methyl (S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4,4-difluoropyrrolidine-2-carboxylate (700 mg, 1.09 mmol). The mixture remained undissolved. Zinc powder (711.41 mg, 10.88 mmol) was added and the mixture was heated in an oil bath preheated to 75°C for 16 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, filtered to remove insoluble matter, and the filter cake was washed with an appropriate amount of methanol. The filtrate was concentrated under reduced pressure. The residue was added with 10 mL of water and extracted with 10 mL x 3 of ethyl acetate. The combined organic phases were washed with 10 mL of brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford the crude title compound (580 mg, 1.05 mmol), which was used in the next step without purification.

[0523] Its structural characterization data are as follows:

[0524] ESI-MS (m / z): 389.2 [M+H] +

[0525] Step 4: Preparation of (S)-8-(benzyloxy)-2,2-difluoro-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-29-5)

[0526] (S)-8-(Benzyloxy)-2,2-difluoro-7-methoxy-1,2,3,11-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (100 mg, 257.49 μmol) was dissolved in THF (2 mL). Under nitrogen, the mixture was cooled to 0°C and borane (1 M in THF, 2.57 mL) was added dropwise. After addition, the mixture was allowed to warm to room temperature and allowed to react for 1.5 hours. After completion of the reaction, the reaction mixture was cooled to 0°C and quenched by the addition of an appropriate amount of methanol. The reaction mixture was stirred at room temperature for 1 hour. 10 mL of water was added to the reaction mixture, and the mixture was extracted with 10 mL of ethyl acetate x 3. The organic phase was washed with 10 mL of brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate-petroleum ether = 10-70%) and concentrated again under reduced pressure to obtain the title compound (23 mg, 61.43 μmol).

[0527] Its structural characterization data are as follows:

[0528] ESI-MS (m / z): 385.1 [M+H] +

[0529] Step 5: Preparation of (S)-2,2-difluoro-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5-one (7-29-6)

[0530] (S)-8-(Benzyloxy)-2,2-difluoro-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (23 mg, 61.43 μmol) was suspended in methanol (2 mL) and ethyl acetate (1 mL) was added to completely dissolve the starting material. Pd / C (7.46 mg, 6.14 μmol, 10% purity) was added, and the hydrogen atmosphere was replaced three times. The reaction was stirred at room temperature under a hydrogen balloon for 3 hours. After completion of the reaction, the reaction mixture was filtered, the filter cake was washed with an appropriate amount of ethyl acetate, and the filtrate was directly concentrated under reduced pressure to obtain the crude title compound (23 mg, 59.88 μmol), which was used directly in the next step without purification.

[0531] Its structural characterization data are as follows:

[0532] ESI-MS (m / z): 285.2 [M+H] +

[0533] Step 6: Preparation of tert-butyl (4-((S)-8-((5-(((S)-2,2-difluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro)-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (7-29-7)

[0534] Tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (20.63 mg, 35.18 μmol) and (S)-2,2-difluoro-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (10 mg, 35.18 μmol) were dissolved in DMF (0.5 mL), K2CO3 (14.59 mg, 105.54 μmol) was added, and the mixture was stirred at room temperature for 18 hours. After the reaction was completed, 4 mL of water was added to the reaction solution, and the mixture was extracted with 5 mL of ethyl acetate × 3. The organic phase was washed with 5 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude title compound (28 mg, 26.59 μmol), which was used in the next step without purification.

[0535] Its structural characterization data are as follows:

[0536] ESI-MS (m / z): 790.4 [M+H] +

[0537] Step 7: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((S)-2,2-difluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-29)

[0538] To tert-butyl (4-((S)-8-((5-(((S)-2,2-difluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro)-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (28 mg, 35.45 μmol) was added dichloromethane (1 mL) and then TFA (0.2 mL) at room temperature. After the addition was complete, the reaction was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure to remove the solvent. The residue was directly purified by preparative HPLC and freeze-dried to obtain the title compound (4.5 mg, 6.29 μmol).

[0539] Its structural characterization data are as follows:

[0540] ESI-MS (m / z): 690.4 [M+H] + ;345.9[M / 2+1] +

[0541] 1 H NMR (400MHz, DMSO-d6) δ7.33(s,1H),7.29(s,1H),7.26(s,1H),7.11(d,J=8.4Hz,1 H),6.53(d,J=8.4Hz,2H),6.49(brs,1H),6.30(s,2H),5.19(brs,1H),4.14-4.10(m ,1H),3.98-3.84(m,7H),3.67(s,3H),3.64(s,3H),3.57-3.50(m,2H),3.25-3.14( m,3H),2.75-2.72(m,2H),1.82-1.78(m,3H),1.82-1.78(m,4H),1.59-1.53(m,2H).

[0542] The preparation method is as follows:

[0543] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0544] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0545] Preparation Example 14: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2S,11aS)-2-fluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-30)

[0546] Step 1: Preparation of (2S,4S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-fluoropyrrolidine-2-carboxylic acid methyl ester (7-30-2)

[0547] Methyl (2S,4S)-4-fluoropyrrolidine-2-carboxylate (1.07 g, 7.25 mmol) and 4-(benzyloxy)-5-methoxy-2-nitrobenzoic acid (2 g, 6.59 mmol) were dissolved in DMF (10 mL). HATU (2.76 g, 7.25 mmol) and DIPEA (2.56 g, 19.78 mmol) were added and reacted at room temperature for 1 hour. The reaction solution was added to ethyl acetate (100 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (ethyl acetate / petroleum ether = 20% to 60%) and concentrated again to obtain the title compound (2.3 g, 5.32 mmol).

[0548] Its structural characterization data are as follows:

[0549] ESI-MS (m / z): 433.2 [M+H] +

[0550] Step 2: Preparation of (2S,11aS)-8-(benzyloxy)-2-fluoro-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-30-3)

[0551] Methyl (2S,4S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-fluoropyrrolidine-2-carboxylate (2.3 g, 5.32 mmol) was dissolved in methanol (50 mL). Zinc powder (3.46 g, 53.19 mmol) was added, followed by a saturated aqueous solution of ammonium chloride (10 mL). The temperature was raised to 85°C and refluxed for 16 hours. The reaction mixture was added to ethyl acetate (150 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude title compound (1.8 g, 4.86 mmol), which was used directly in the next step without purification.

[0552] Its structural characterization data are as follows:

[0553] ESI-MS (m / z): 371.2 [M+H] +

[0554] Step 3: Preparation of (2S,11aS)-8-(benzyloxy)-2-fluoro-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-30-4)

[0555] Dissolve (2S,11aS)-8-(benzyloxy)-2-fluoro-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (1.8 g, 4.86 mmol) in tetrahydrofuran (100 mL). Add sodium borohydride (1064.34 mg, 14.58 mmol). Then, add trifluoroacetic acid (1108.1 mg, 9.72 mmol) dropwise at room temperature until no significant gas escapes. The mixture is heated to 75°C and refluxed for 5 hours. The reaction mixture is concentrated, water (50 mL) is added, and the mixture is extracted twice with ethyl acetate (50 mL x 2). The combined organic phases are washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude title compound (1.4 g, 3.93 mmol), which is used directly in the next step without purification.

[0556] Its structural characterization data are as follows:

[0557] ESI-MS (m / z): 357.3 [M+H] +

[0558] Step 4: Preparation of (2S,11aS)-2-fluoro-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-30-5)

[0559] Dissolve (2S,11aS)-8-(benzyloxy)-2-fluoro-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (200 mg, 561.18 μmol) in methanol (10 mL). Add palladium on carbon (5 mg) and react under a hydrogen atmosphere for 1 hour. After completion, filter the reaction, dry the filtrate over anhydrous sodium sulfate, filter, and concentrate to obtain the crude title compound (0.14 g, 525.79 μmol), which is used directly in the next step without purification.

[0560] Its structural characterization data are as follows:

[0561] ESI-MS (m / z): 267.1 [M+H] +

[0562] Step 5: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2S,11aS)-2-fluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-30-6)

[0563] (2S,11aS)-2-fluoro-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (10 mg, 37.56 μmol) and (S)-tert-butyl(4-(8-((5-bromopentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (22.03 mg, 37.56 μmol) were dissolved in DMF (1 mL). Potassium carbonate (15.55 mg, 112.67 μmol) was added and the mixture was stirred for 16 hours. The reaction was quenched with water and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) was followed by further concentration to afford the title compound (20 mg, 25.91 μmol).

[0564] Its structural characterization data are as follows:

[0565] ESI-MS (m / z): 772.3 [M+H] +

[0566] Step 6: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2S,11aS)-2-fluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-30)

[0567] (S)-2-(4-Aminophenyl)-8-((5-(((2S,11aS)-2-fluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (20 mg, 25.91 μmol) was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (0.5 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. After concentration under reduced pressure, purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%) and lyophilization, the title compound (8.14 mg, 12.00 μmol) was obtained.

[0568] Its structural characterization data are as follows:

[0569] ESI-MS (m / z): 673.3 [M+H] +

[0570] 1 H NMR (400 MHz, DMSO) δ 7.40 (s, 1H), 7.29 (s, 1H), 7.24 (s, 1H), 6.87-6.70 (m, 2H), 6.60-6.45 (m, 2H), 6.31 (s, 2H), 4.17 (s, 1H), 4.03-3.87 (m, 4H), 3.87-3.76 (m, 2H), 3.67 (s, 3H), 3.65 (s, 3H), 3.58-3.45 (m, 4H), 3.29-3.20 (m, 2H), 2.82-2.61 (m, 2H), 2.20-2.06 (m, 2H), 1.86-1.73 (m, 4H), 1.60-1.53 ​​(m, 2H). The preparation method is as follows:

[0571] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0572] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0573] Preparation Example 15: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2R,11aS)-2-fluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-31)

[0574] Step 1: Preparation of (2R,4S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-fluoropyrrolidine-2-carboxylic acid methyl ester (7-31-2)

[0575] Methyl (2R,4S)-4-fluoropyrrolidine-2-carboxylate (1.1 g, 7.25 mmol, FR) and 4-(benzyloxy)-5-methoxy-2-nitrobenzoic acid (2.05 g, 6.59 mmol, FR) were dissolved in DMF (10 mL). HATU (2.76 g, 7.25 mmol) and DIPEA (2.56 g, 19.78 mmol) were added and reacted at room temperature for 1 hour. The reaction solution was added to ethyl acetate (100 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (ethyl acetate / petroleum ether = 20% to 60%) and concentrated again to obtain the title compound (2.1 g, 4.85 mmol).

[0576] Its structural characterization data are as follows:

[0577] ESI-MS (m / z): 433.2 [M+H] +

[0578] Step 2: Preparation of (2R,11aS)-8-(benzyloxy)-2-fluoro-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-31-3)

[0579] Methyl (2R,4S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-fluoropyrrolidine-2-carboxylate (2.1 g, 4.85 mmol) was dissolved in methanol (50 mL). Zinc powder (3.4 g, 53.1 mmol) was added, followed by a saturated aqueous solution of ammonium chloride (10 mL). The mixture was heated to 85°C and refluxed for 16 hours. The reaction mixture was added to ethyl acetate (150 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude title compound (1.7 g, 4.7 mmol), which was used directly in the next step without purification.

[0580] Its structural characterization data are as follows:

[0581] ESI-MS (m / z): 371.2 [M+H] +

[0582] Step 3: Preparation of (2R,11aS)-8-(benzyloxy)-2-fluoro-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-31-4)

[0583] Dissolve (2R,11aS)-8-(benzyloxy)-2-fluoro-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (400 mg, 1.04 mmol) in tetrahydrofuran (5 mL) and cool to 0°C. Then, add tetrahydrofuran borane complex (10.3 mL) dropwise. After addition, warm to room temperature and react for 3 hours. Add methanol (10 mL) dropwise to the reaction solution, then concentrate to obtain the crude title compound. Purify on a silica gel column (ethyl acetate / petroleum ether = 0% to 70%) and concentrate to obtain the title compound (110 mg, 0.31 mmol).

[0584] Its structural characterization data are as follows:

[0585] ESI-MS (m / z): 357.3 [M+H] +

[0586] Step 4: Preparation of (2R,11aS)-2-fluoro-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-31-5)

[0587] Dissolve (2S,11aS)-8-(benzyloxy)-2-fluoro-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (110 mg, 0.31 mmol) in tetrahydrofuran (5 mL). Add palladium on carbon (10 mg). The mixture is reacted under a hydrogen atmosphere for 1 hour. After completion, the reaction is filtered through celite, and the filtrate is dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude title compound (50 mg, 190.1 μmol), which is used directly in the next step without purification.

[0588] Its structural characterization data are as follows:

[0589] ESI-MS (m / z): 267.1 [M+H] +

[0590] Step 5: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2R,11aS)-2-fluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-31-6)

[0591] (2R,11aS)-2-Fluoro-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (50 mg, 190.1 μmol) and tert-butyl (S)-(4-(8-((5-bromopentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (117.3 mg, 199.5 μmol) were dissolved in DMF (3 mL). Potassium carbonate (52.4 mg, 380 μmol) was added and the mixture was stirred for 16 hours. The reaction was quenched by adding water and extracted with ethyl acetate three times (20 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) was followed by further concentration to afford the title compound (88 mg, 114.1 μmol).

[0592] Its structural characterization data are as follows:

[0593] ESI-MS (m / z): 772.3 [M+H] +

[0594] Step 6: Preparation of (S)-2-(4-aminophenyl)-8-((5-(((2R,11aS)-2-fluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-31)

[0595] (S)-2-(4-Aminophenyl)-8-((5-(((2R,11aS)-2-fluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (88 mg, 114.1 μmol)) was dissolved in dichloromethane (3 mL). Trifluoroacetic acid (1.5 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. After concentration under reduced pressure, purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%) and lyophilization, the title compound (8.0 mg, 11.7 μmol) was obtained.

[0596] Its structural characterization data are as follows:

[0597] ESI-MS (m / z): 673.3 [M+H] +

[0598] 1 H NMR (400MHz, DMSO): δ7.39 (s, 1H), 7.29 (s, 2H), 7.14 (d, J = 7.8Hz, 2H), 6.58 (d, J = 7. 4Hz,2H),6.29(d,J=10.0Hz,2H),5.26(d,J=53.6Hz,1H),4.21-4.08(m,1H),4.01-3 .89(m,4H),3.85-3.76(m,2H),3.65(d,J=5.8Hz,6H),3.60-3.49(m,4H),3.29-3.11 (m,4H),3.04-2.95(m,1H),2.74-2.63(m,1H),1.88-1.73(m,4H),1.62-1.50(m,2H).

[0599] The preparation method is as follows:

[0600] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0601] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0602] Preparation Example 16: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((2S,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-32)

[0603] Step 1: Preparation of methyl (2S,4S)-4-(trifluoromethyl)pyrrolidine-2-carboxylate (7-32-2)

[0604] Dissolve 1-(tert-Butyl)-2-methyl(2S,4S)-4-(trifluoromethyl)pyrrolidine-1,2-dicarboxylic acid (1.0 g, 3.53 mmol, FR) in methanol (10 mL). Add HCl (4M in EA) (5 mL) and react at room temperature for 1 hour. Concentrate to obtain the crude product, which is used in the next step without purification (820.79 mg, 3.51 mmol).

[0605] Its structural characterization data are as follows:

[0606] ESI-MS (m / z): 234.2 [M+H] +

[0607] Step 2: Preparation of (2R,4S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-(trifluoromethyl)pyrrolidine-2-carboxylic acid methyl ester (7-32-3)

[0608] Methyl (2S,4S)-4-(trifluoromethyl)pyrrolidine-2-carboxylate (820.79 mg, 3.51 mmol, CL) and 4-(benzyloxy)-5-methoxy-2-nitrobenzoic acid (1.18 g, 3.88 mmol, FR) were dissolved in DMF (10 mL). HATU (1.48 g, 3.88 mmol) and DIPEA (1.36 g, 10.59 mmol) were added and reacted at room temperature for 1 hour. The reaction solution was added to ethyl acetate (100 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was purified on a silica gel column (ethyl acetate / petroleum ether = 20% to 60%) and concentrated again to obtain the title compound (1.5 g, 3.1 mmol).

[0609] Its structural characterization data are as follows:

[0610] ESI-MS (m / z): 483.2 [M+H] +

[0611] Step 3: Preparation of (2S,11aS)-8-(benzyloxy)-7-methoxy-2-(trifluoromethyl)-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-32-4)

[0612] Methyl (2S,4R)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-(trifluoromethyl)pyrrolidine-2-carboxylate (1.5 g, 3.1 mmol) was dissolved in methanol (30 mL). Zinc powder (2.2 g, 35.1 mmol) was added, followed by a saturated aqueous solution of ammonium chloride (40 mL). The mixture was heated to 85°C and refluxed for 16 hours. The reaction mixture was added to ethyl acetate (150 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude title compound (1.4 g, 3.3 mmol), which was used directly in the next step without purification.

[0613] Its structural characterization data are as follows:

[0614] ESI-MS (m / z): 421.2 [M+H] +

[0615] Step 4: Preparation of (2S,11aS)-8-(benzyloxy)-7-methoxy-2-(trifluoromethyl)-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-32-5)

[0616] Dissolve (2S,11aS)-8-(benzyloxy)-7-methoxy-2-(trifluoromethyl)-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (400 mg, 0.95 mmol) in tetrahydrofuran (5 mL) and cool to 0°C. Then, add tetrahydrofuran borane complex (10.3 mL) dropwise. After addition, warm to room temperature and react for 3 hours. Add methanol (10 mL) dropwise to the reaction solution, then concentrate to obtain the crude title compound. Purify on a silica gel column (ethyl acetate / petroleum ether = 0% to 70%) and concentrate to obtain the title compound (100 mg, 0.24 mmol).

[0617] Its structural characterization data are as follows:

[0618] ESI-MS (m / z): 408.1 [M+H] +

[0619] Step 5: Preparation of (2S,11aS)-8-hydroxy-7-methoxy-2-(trifluoromethyl)-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-32-6)

[0620] Dissolve (2S,11aS)-8-(benzyloxy)-7-methoxy-2-(trifluoromethyl)-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (100 mg, 0.24 mmol) in tetrahydrofuran (3 mL). Add palladium on carbon (10 mg). The mixture is reacted under a hydrogen atmosphere for 1 hour. After completion, the reaction is filtered, and the filtrate is dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude title compound (70 mg, 0.22 mmol), which is used directly in the next step without purification.

[0621] Its structural characterization data are as follows:

[0622] ESI-MS (m / z): 317.1 [M+H] +

[0623] Step 6: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((2S,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-32-7)

[0624] (2R,11aS)-8-Hydroxy-7-methoxy-2-(trifluoromethyl)-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (70 mg, 0.22 mmol) and (S)-tert-butyl(4-(8-((5-bromopentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (136 mg, 0.23 mmol) were dissolved in DMF (2 mL). Potassium carbonate (61 mg, 0.44 mmol) was added and the mixture was stirred for 16 hours. The reaction was quenched with water and extracted three times with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. Purification was performed on a silica gel column (MeOH / DCM = 0% to 5%) and further concentrated to afford the title compound (72 mg, 88.3 μmol).

[0625] Its structural characterization data are as follows:

[0626] ESI-MS (m / z): 822.5 [M+H] +

[0627] Step 7: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((2S,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-32)

[0628] (S)-2-(4-Aminophenyl)-7-methoxy-8-((5-(((2S,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (72 mg, 88.3 μmol) was dissolved in dichloromethane (3 mL). Trifluoroacetic acid (1.5 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. After concentration under reduced pressure, purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%) and lyophilization, the title compound (9.8 mg, 13.6 μmol) was obtained.

[0629] Its structural characterization data are as follows:

[0630] ESI-MS (m / z): 723.3 [M+H] +

[0631] 1 H NMR (400MHz, DMSO): δ7.41(s,1H),7.27(d,J=12.2Hz,2H),7.11(d,J=8.2Hz,2H),6.5 3(d,J=8.4Hz,2H),6.29(d,J=12.4Hz,2H),5.20(s,2H),4.19-4.08(m,1H),3.97-3.8 8(m,4H),3.87-3.76(m,2H),3.66(s,3H),3.64(s,3H),3.59-3.46(m,4H),3.28-3.16 (m,4H),3.04-2.94(m,1H),2.73-2.64(m,1H),1.84-1.76(m,4H),1.61-1.50(m,2H).

[0632] The preparation method is as follows:

[0633] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0634] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0635] Preparation Example 17: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((2R,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-33)

[0636] Step 1: Preparation of methyl (2S,4R)-4-(trifluoromethyl)pyrrolidine-2-carboxylate (7-33-2)

[0637] Dissolve 1-(tert-butyl)-2-methyl(2S,4R)-4-(trifluoromethyl)pyrrolidine-1,2-dicarboxylic acid (1.0 g, 3.53 mmol) in methanol (10 mL), add HCl (4M in EA) (5 mL), and react at room temperature for 1 hour. Concentrate to obtain the crude product, which is used directly in the next step without purification (824.79 mg, 3.53 mmol).

[0638] Its structural characterization data are as follows:

[0639] ESI-MS (m / z): 234.2 [M+H] +

[0640] Step 2: Preparation of (2S,4R)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-(trifluoromethyl)pyrrolidine-2-carboxylic acid methyl ester (7-33-3)

[0641] Methyl (2S,4R)-4-(trifluoromethyl)pyrrolidine-2-carboxylate (824.79 mg, 3.53 mmol) and 4-(benzyloxy)-5-methoxy-2-nitrobenzoic acid (1.18 g, 3.88 mmol) were dissolved in DMF (10 mL). HATU (1.48 g, 3.88 mmol) and DIPEA (1.36 g, 10.59 mmol) were added and reacted at room temperature for 1 hour. The reaction solution was added to ethyl acetate (100 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (ethyl acetate / petroleum ether = 20% to 60%) and concentrated again to obtain the title compound (1.7 g, 3.52 mmol).

[0642] Its structural characterization data are as follows:

[0643] ESI-MS (m / z): 483.2 [M+H] +

[0644] Step 3: Preparation of (2R,11aS)-8-(benzyloxy)-7-methoxy-2-(trifluoromethyl)-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-33-4)

[0645] Methyl (2S,4R)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-(trifluoromethyl)pyrrolidine-2-carboxylate (1.7 g, 3.52 mmol) was dissolved in methanol (30 mL). Zinc powder (2.29 g, 35.24 mmol) was added, followed by a saturated aqueous solution of ammonium chloride (40 mL). The mixture was heated to 85°C and refluxed for 16 hours. The reaction mixture was added to ethyl acetate (150 mL) and saturated brine (100 mL), stirred, and allowed to stand for separation. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude title compound (1.48 g, 3.52 mmol), which was used directly in the next step without purification.

[0646] Its structural characterization data are as follows:

[0647] ESI-MS (m / z): 421.2 [M+H] +

[0648] Step 4: Preparation of (2R,11aS)-8-(benzyloxy)-7-methoxy-2-(trifluoromethyl)-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-33-5)

[0649] (2R,11aS)-8-(benzyloxy)-7-methoxy-2-(trifluoromethyl)-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (1.48 g, 3.52 mmol) was dissolved in tetrahydrofuran (30 mL). Sodium borohydride (770.88 mg, 10.56 mmol) was added, followed by the dropwise addition of trifluoroacetic acid (802.56 mg, 7.04 mmol) at room temperature until the gas ceased to escape. The mixture was then heated to 75°C and refluxed for 5 hours. The reaction mixture was concentrated, water (50 mL) was added, and the mixture was extracted twice with ethyl acetate (50 mL x 2). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude title compound (1.1 g, 2.71 mmol), which was used directly in the next step without further purification.

[0650] Its structural characterization data are as follows:

[0651] ESI-MS (m / z): 408.1 [M+H] +

[0652] Step 5: Preparation of (2R,11aS)-8-hydroxy-7-methoxy-2-(trifluoromethyl)-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-33-6)

[0653] Dissolve (2R,11aS)-8-(benzyloxy)-7-methoxy-2-(trifluoromethyl)-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (100 mg, 246.06 μmol) in methanol (10 mL). Add palladium on carbon (5 mg) and react under a hydrogen atmosphere for 1 hour. After completion, filter the reaction, dry the filtrate over anhydrous sodium sulfate, filter, and concentrate to obtain the crude title compound (77.82 mg, 246.05 μmol), which is used directly in the next step without purification.

[0654] Its structural characterization data are as follows:

[0655] ESI-MS (m / z): 317.1 [M+H] +

[0656] Step 6: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((2R,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-33-7)

[0657] (2R,11aS)-8-hydroxy-7-methoxy-2-(trifluoromethyl)-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (10.78 mg, 34.10 μmol) and (S)-tert-butyl(4-(8-((5-bromopentyl)oxy)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl)phenyl)carbamate (20.00 mg, 34.10 μmol) were dissolved in DMF (1 mL). Potassium carbonate (14.12 mg, 102.30 μmol) was added and the mixture was stirred for 16 hours. The reaction was quenched by adding water and extracted with ethyl acetate three times (10 mL x 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. Purification on a silica gel column (MeOH / DCM = 0% to 5%) was followed by further concentration to afford the title compound (20 mg, 24.33 μmol).

[0658] Its structural characterization data are as follows:

[0659] ESI-MS (m / z): 822.5 [M+H] +

[0660] Step 7: Preparation of (S)-2-(4-aminophenyl)-7-methoxy-8-((5-(((2R,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-33)

[0661] (S)-2-(4-Aminophenyl)-7-methoxy-8-((5-(((2R,11aS)-7-methoxy-5-oxo-2-(trifluoromethyl)-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (20 mg, 24.33 μmol) was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (0.5 mL) was added dropwise with stirring. After the addition was complete, stirring was continued for 1 hour. After concentration under reduced pressure, purification by reverse phase column chromatography (acetonitrile / 1% formic acid aqueous solution = 0% to 70%) and lyophilization, the title compound (9.33 mg, 12.80 μmol) was obtained.

[0662] Its structural characterization data are as follows:

[0663] ESI-MS (m / z): 723.3 [M+H] +

[0664] 1 H NMR(400MHz,DMSO)δ7.39(s,1H),7.28(s,1H),7.25(s,1H),6.82(s,2H),6. 59-6.43(m,2H),6.31(s,2H),4.18-4.08(m,1H),3.97-3.85(m,4H),3.82-3. 74(m,2H),3.68(s,3H),3.66(s,3H),3.56-3.44(m,4H),3.28-3.19(m,2H), 2.81-2.65(m,2H),2.21-2.04(m,2H),1.87-1.70(m,4H),1.59-1.52(m,2H).

[0665] The preparation method is as follows:

[0666] Chromatographic column: SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0667] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0668] Preparation Examples 18 and 19: (S)-8-((5-(((2S,11aS)-2-(4-aminophenyl)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7- Preparation of (S)-8-((5-(((2R,11aS)-2-(4-aminophenyl)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-35)

[0669] (S)-8-((5-(((S)-2-(4-aminophenyl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-ethoxy-1,10,11,11-tetrahydro-3H,5H-spiro[e]pyrrolo[1,2-a][1,4]azepine-2,1'-cyclopropane]-5-one (Compound 7-5) (20 mg, 29.42 μmol) was dissolved in a mixed solvent of methanol (5 mL) and tetrahydrofuran (5 mL), 10% palladium on carbon (2 mg) was added, and the mixture was replaced with hydrogen and stirred under protection for 2 hours. The reaction solution was filtered through a pad of celite, and the filtrate was concentrated under reduced pressure and purified by preparative high performance liquid chromatography and freeze-dried to obtain compound 7-34 (2.77 mg, 3.13 μmol) and compound 7-35 (5.66 mg, 6.69 μmol).

[0670] Its structural characterization data are as follows:

[0671] MS m / z(ESI): doublet 682.5[M+H] +

[0672] Compound 7-34: 1H NMR (400MHz, DMSO) δ7.40-7.24(m,4H),7.10(d,J=8.3Hz,2H),6.31(t,J=13.0Hz,2H),3.97(s ,1H),3.92(t,J=6.2Hz,4H),3.83(d,J=7.6Hz,2H),3.65(d,J=5.1Hz,6H),3.55-3.47(m,4H), 3.30(dd,J=19.7,10.5Hz,2H),3.26-3.11(m,2H),2.31(dd,J=12.5,7.5Hz,1H),2.17-2.09(m ,1H),1.96(dd,J=12.4,7.2Hz,1H),1.86-1.72(m,5H),1.60-1.51(m,2H),0.68-0.48(m,4H).

[0673] Compound 7-35: 1 H NMR (400MHz, DMSO) δ7.35(dd,J=18.7,10.3Hz,4H),7.11(d,J=8.2Hz,2H),6.29(d,J=15.3Hz,2H ),4.05(d,J=3.6Hz,1H),3.92(t,J=6.4Hz,4H),3.83(s,2H),3.65(t,J=2.7Hz,6H),3.53(d,J=11 .8Hz,4H),3.33(d,J=11.7Hz,2H),3.12(ddd,J=34.2,13.2,8.2Hz,2H),2.46(d,J=5.1Hz,1H),2 .00-1.93(m,1H),1.87-1.70(m,6H),1.56(d,J=6.6Hz,2H),0.58(ddt,J=15.7,12.7,6.9Hz,4H).

[0674] Its preparation method is as follows:

[0675] Chromatographic column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0676] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% trifluoroacetic acid)

[0677] Preparation Example 20: Preparation of (S)-8-((5-(((S)-2-(Furan-3-yl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-36)

[0678] Step 1: Preparation of (S)-8-((5-bromopentyl)oxy)-2-(furan-3-yl)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-36-1)

[0679] Furan-3-ylboronic acid (40.15 mg, 358.85 μmol), (S)-8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl trifluoromethanesulfonate (100 mg, 179.43 μmol), tetrakis(triphenylphosphine)palladium (20.73 mg, 17.94 μmol) and sodium carbonate (76.07 mg, 717.70 μmol) were mixed, and water (0.5 mL), toluene (2 mL) and ethanol (0.5 mL) were added. The atmosphere was then purged with nitrogen three times and the mixture was heated at 80°C for 3 hours. After completion of the reaction, the reaction mixture was added with 10 mL of water and extracted with 5 mL x 3 of ethyl acetate. The organic phase was washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0% to 70%) and concentrated again under reduced pressure to obtain the title compound (60 mg, 126.23 μmol).

[0680] Its structural characterization data are as follows:

[0681] ESI-MS (m / z): 475.1 [M+H] +

[0682] Step 2: Preparation of (S)-2-(furan-3-yl)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-36-2)

[0683] (S)-8-((5-bromopentyl)oxy)-2-(furan-3-yl)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (60 mg, 126.23 μmol) and (S)-8-hydroxy-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one (34.63 mg, 126.23 μmol) were dissolved in DMF (1 mL) and potassium carbonate (52.34 mg, 378.68 μmol) was added. The reaction was stirred at room temperature for 20 hours. After completion of the reaction, 10 mL of water was added to the reaction mixture, causing a large amount of solid to precipitate. The mixture was then extracted with 10 mL x 3 of a 10:1 mixture of ethyl acetate and methanol. The organic phase was washed with 10 mL of brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (DCM:MeOH = 8:1) and concentrated again under reduced pressure to afford the title compound (43 mg, 50.80 μmol).

[0684] Its structural characterization data are as follows:

[0685] ESI-MS (m / z): 669.3 [M+H] +

[0686] Step 3: Preparation of (S)-8-((5-(((S)-2-(furan-3-yl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-36)

[0687] (S)-2-(Furan-3-yl)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (24 mg, 35.89 μmol) was dissolved in THF (2 mL), NaBH4 (12.22 mg, 323.00 μmol) was added, and then TFA (24.55 mg, 215.33 μmol) was added dropwise. A lot of bubbles were generated in the system. After the bubbles disappeared, transfer the mixture to an oil bath and heat to 75°C for 22 hours. After completion, the reaction mixture was extracted three times with 5 mL of water and 5 mL of ethyl acetate. The organic phase was washed with 5 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by preparative HPLC and freeze-dried to yield the title compound (6.56 mg, 9.77 μmol).

[0688] Its structural characterization data are as follows:

[0689] ESI-MS (m / z): 655.4 [M+H] +

[0690] 1 H NMR (400MHz, DMSO-d6) δ7.66(d,J=10.0Hz,2H),7.32(s,2H),7.28(s,1H),6.85(s,1H),6.53(d,J=6 .4Hz,1H),6.31-6.26(m,3H),4.20-4.13(m,1H),3.98-3.88(m,4H),3.88-3.70(m,1H),3.66(s,3H) ,3.64(s,3H),3.58-3.43(m,4H),3.25-3.11(m,4H),2.62(dd,J=16.8,4.4Hz,1H),1.95(dd,J=12.4 ,7.2Hz,1H),1.79(dd,J=12.4,7.2Hz,5H),1.61-1.49(m,2H),0.68-0.61(m,1H),0.60-0.48(m,3H).

[0691] The preparation method is as follows:

[0692] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0693] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0694] Preparation Example 21: Preparation of (S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-2-(thien-3-yl)-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-37)

[0695] Step 1: Preparation of (S)-8-((5-bromopentyl)oxy)-2-(furan-3-yl)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-37-1)

[0696] Thiophene-3-ylboronic acid (91.84 mg, 717.70 μmol), (S)-8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl trifluoromethanesulfonate (200 mg, 358.85 μmol), tetrakis(triphenylphosphine)palladium (41.47 mg, 35.89 μmol), and sodium carbonate (152.14 mg, 1.44 mmol) were mixed and added with water (1 mL), toluene (4 mL), and ethanol (1 mL). The mixture was then purged with nitrogen three times and heated to 80°C for 2 hours. After completion of the reaction, the reaction mixture was added with 10 mL of water and extracted three times with 5 mL of ethyl acetate. The organic phase was washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0% to 70%) and concentrated again under reduced pressure to give the title compound (58 mg, 118.03 μmol).

[0697] Its structural characterization data are as follows:

[0698] ESI-MS (m / z): 491.1 [M+H] +

[0699] Step 2: Preparation of (S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo)[1,2-a][1,4]diazepine-2,1'-cyclopropyl]-8-yl)oxy)pentyl)oxy)-2-(thiophen-3-yl)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-37-2)

[0700] (S)-8-((5-bromopentyl)oxy)-2-(furan-3-yl)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (55 mg, 111.93 μmol) and (S)-8-hydroxy-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one (30.70 mg, 111.93 μmol) were dissolved in DMF (2 mL) and potassium carbonate (46.41 mg, 335.78 μmol) was added. The reaction was stirred at room temperature for 40 hours. After the reaction was completed, the reaction solution was filtered to remove insoluble matter, and the filtrate was purified by preparative HPLC and freeze-dried to obtain the title compound (15 mg, 21.90 μmol).

[0701] Its structural characterization data are as follows:

[0702] ESI-MS (m / z): 685.3 [M+H] +

[0703] The preparation method is as follows:

[0704] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0705] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0706] Step 3: Preparation of (S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-2-(thien-3-yl)-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-37)

[0707] (S)-7-Methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropyl]-8-yl)oxy)pentyl)oxy)-2-(thiophen-3-yl)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (15 mg, 21.90 μmol) was dissolved in THF (2 mL). Sodium borohydride (8.29 mg, 219.04 μmol) was added, resulting in slight bubbling. TFA (14.99 mg, 131.43 μmol) was added dropwise, resulting in significant bubbling. The reaction was heated to 70°C and allowed to react for 24 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, further purified by preparative HPLC and freeze-dried to obtain the title compound (2.83 mg, 4.01 μmol).

[0708] Its structural characterization data are as follows:

[0709] ESI-MS (m / z): 671.3 [M+H] +

[0710] 1 H NMR (400MHz, DMSO-d6) δ7.53(dd,J=4.8,2.8Hz,1H),7.45(dd,J=3.6,1.2Hz,2H),7.32(s,1H),7.29-7.28(m,2H),6.55(d,J=6. 0Hz,1H),6.31(s,1H),6.30-6.26(m,2H),4.21-4.15(m,1H),3.93(dd,J=13.2,6.6Hz,4H),3.83(dd,J=15.2,7.6Hz,1H),3.66(s ,3H),3.64(s,3H),3.55-3.50(m,2H),3.50-3.44(m,1H),3.24(dd,J=13.2,10.0Hz,2H),3.14(dd,J=12.0,8.8Hz,1H),2.74(dd ,J=16.4,4.0Hz,1H),2.03-1.90(m,2H),1.79(dd,J=12.4,7.6Hz,5H),1.61-1.50(m,2H),0.68-0.61(m,1H),0.61-0.48(m,3H).

[0711] The preparation method is as follows:

[0712] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0713] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0714] Preparation Example 22: Preparation of (S)-8-((5-(((S)-2-(5-(hydroxymethyl)thiophen-3-yl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-38)

[0715] Step 1: Preparation of (S)-8-((5-bromopentyl)oxy)-2-(5-(hydroxymethyl)thiophen-3-yl)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-38-1)

[0716] (5-(Hydroxymethyl)thiophen-3-yl)boronic acid (113.38 mg, 717.70 μmol), (S)-8-((5-bromopentyl)oxy)-7-methoxy-5,11-dioxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-2-yl trifluoromethanesulfonate (200 mg, 358.85 μmol), tetrakis(triphenylphosphine)palladium (41.47 mg, 35.89 μmol), and sodium carbonate (152.14 mg, 1.44 mmol) were mixed, and water (1 mL), toluene (4 mL), and ethanol (1 mL) were added. The mixture was then purged with nitrogen three times and heated to 80°C for 2 hours. After completion of the reaction, 10 mL of water was added to the reaction mixture, and the mixture was extracted three times with 5 mL of ethyl acetate. The organic phase was washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0% to 100%) and concentrated again under reduced pressure to obtain the title compound (30 mg, 50.06 μmol).

[0717] Its structural characterization data are as follows:

[0718] ESI-MS (m / z): 521.1 [M+H] +

[0719] Step 2: Preparation of (S)-2-(5-(hydroxymethyl)thiophen-3-yl)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-38-2)

[0720] (S)-8-((5-bromopentyl)oxy)-2-(5-(hydroxymethyl)thiophen-3-yl)-7-methoxy-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (30 mg, 57.53 μmol) and (S)-8-hydroxy-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one (15.78 mg, 57.53 μmol) were dissolved in DMF (1 mL) and potassium carbonate (23.85 mg, 172.60 μmol) was added. The mixture was stirred at room temperature for 40 hours. After the reaction was completed, the reaction solution was filtered to remove insoluble matter, and the filtrate was purified by preparative HPLC and freeze-dried to obtain the title compound (10 mg, 13.99 μmol).

[0721] Its structural characterization data are as follows:

[0722] ESI-MS (m / z): 715.3 [M+H] +

[0723] The preparation method is as follows:

[0724] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0725] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0726] Step 3: Preparation of (S)-8-((5-(((S)-2-(5-(hydroxymethyl)thiophen-3-yl)-7-methoxy-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepin-2,1'-cyclopropane]-5-one (7-38)

[0727] (S)-2-(5-(Hydroxymethyl)thiophen-3-yl)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-5,10,11,11a)-tetrahydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (10 mg, 13.99 μmol) was dissolved in THF (2 mL). Sodium borohydride (5.15 mg, 139.89 μmol) was added with slight bubbling. TFA (9.57 mg, 83.94 μmol) was added dropwise. The temperature was raised to 70°C and the reaction was continued for 24 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was dissolved in methanol, purified by preparative HPLC, and freeze-dried to obtain the title compound (2.18 mg, 2.96 μmol).

[0728] Its structural characterization data are as follows:

[0729] ESI-MS (m / z): 701.3 [M+H] +

[0730] 1 H NMR (400MHz, DMSO-d6) δ7.35(s,1H),7.32(s,1H),7.28(s,1H),7.24(s,1H),7.15(d,J=1.2Hz,1H),6.55(d,J=6.4Hz,1H ),6.30(d,J=10.8Hz,3H),5.48(s,1H),4.60(d,J=3.6Hz,2H),4.17(dd,J=15.2,10.0Hz,1H),3.93(dd,J=12.8,6.4Hz,4 H),3.83(dd,J=14.8,7.2Hz,1H),3.65(d,J=8.4Hz,6H),3.55-3.46(m,4H),3.29-3.21(m,2H),3.21-3.09(m,2H),2.71( dd,J=16.8,4.0Hz,1H),2.03-1.91(m,2H),1.81-1.75(m,,5H),1.59-1.51(m,2H),0.68-0.61(m,1H),0.60-0.48(m,3H).

[0731] The preparation method is as follows:

[0732] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0733] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0734] Preparation Example 23: Preparation of (S)-8-((5-(((S)-2,2-difluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-39)

[0735] Step 1: Preparation of allyl (S)-8-((5-(((S)-2,2-difluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate (7-39-1)

[0736] (S)-2,2-Difluoro-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (19 mg, 66.84 μmol) and (S)-8-((5-bromopentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (39.13 mg, 66.84 μmol) were dissolved in DMF (0.5 mL) and potassium carbonate (27.71 mg, 200.52 μmol) was added. The mixture was stirred at room temperature for 24 hours. Potassium iodide (11.10 mg, 66.84 μmol) was added, and stirring was continued at room temperature for 48 hours. After completion of the reaction, the reaction solution was added with 5 mL of water and extracted with 5 mL of ethyl acetate x 3. The organic phase was washed with 5 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude title compound (70 mg, 64.51 μmol), which was used directly in the next reaction without purification.

[0737] Its structural characterization data are as follows:

[0738] ESI-MS (m / z): 790.4 [M+H] +

[0739] Step 2: Preparation of (S)-8-((5-(((S)-2,2-difluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-39)

[0740] (S)-8-((5-(((S)-2,2-difluoro-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-10(5H)-carboxylic acid allyl ester (70 mg, 88.74 μmol) was dissolved in DMF (1 mL), tetrahydropyrrole (12.62 mg, 177.48 μmol) and tetrakistriphenylphosphine palladium (10.25 mg, 8.87 μmol) were added, and the mixture was purged with nitrogen three times. The reaction was stirred at room temperature under nitrogen for 3 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by preparative high performance liquid chromatography and freeze-dried to obtain the title compound (9.01 mg, 12.66 μmol).

[0741] Its structural characterization data are as follows:

[0742] ESI-MS (m / z): 705.3 [M+H] +

[0743] 1H NMR(400MHz,DMSO-d6)δ7.45(s,1H),7.39(s,1H),7.37(s,1H),7.34(s,1H),7. 29(s,1H),6.91(d,J=8.8Hz,2H),6.56(d,J=6.0Hz,1H),6.49(d,J=6.0Hz,1H), 6.31(d,J=4.8Hz,1H),4.22-4.16(m,1H),3.98-3.86(m,7H),3.75(s,3H),3.66 (s,3H),3.64(s,3H),3.57-3.52(m,2H),3.31-3.14(m,3H),2.78-2.68(m,2H), 2.43-2.29(m,1H),1.82-1.77(m,4H),1.59 -1.52(m,2H).

[0744] The preparation method is as follows:

[0745] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0746] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0747] Preparation Example 24: Preparation of (S)-8-((5-(((2R,11aS)-2-hydroxy-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-40)

[0748] Step 1: Preparation of (S)-8-((5-(((2R,11aS)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (7-40-1)

[0749] (S)-8-((5-bromopentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (45 mg, 76.86 μmol) and (2R,11aS)-2-((tert-butyldimethylsilyl)oxy)-8-hydroxy-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5-one (34.91 mg, 92.23 μmol) were dissolved in DMF (1 mL) and potassium carbonate (21.24 mg, 153.72 μmol) was added. The reaction was stirred for 16 hours. Water was added to precipitate the solid, which was filtered, washed with water, and dried to obtain a crude product (65 mg, 73.60 μmol), which was used in the next step without purification.

[0750] Its structural characterization data are as follows:

[0751] MS m / z(ESI):883.1[M+H] +

[0752] Step 2: Preparation of (S)-8-((5-(((2R,11aS)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-40-2)

[0753] (S)-8-((5-(((2R,11aS)-2-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,1 Allyl 1a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate (65 mg, 73.60 μmol) was dissolved in DMF (1 mL). Tetrakis(triphenylphosphine)palladium (8.51 mg, 7.36 μmol) and tetrahydropyrrole (5.23 mg, 73.60 μmol) were added. The mixture was purged with nitrogen and stirred for 12 hours. Ethyl acetate and water were added, stirred, filtered, and allowed to stand for separation. The organic phase was washed three times with saturated brine, dried, and concentrated under reduced pressure to obtain the crude product (58 mg, 73.60 μmol), which was used directly in the next step without purification.

[0754] Its structural characterization data are as follows:

[0755] MS m / z(ESI):800.5[M+H] +

[0756] Step 3: Preparation of (S)-8-((5-(((2R,11aS)-2-hydroxy-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-40)

[0757] (S)-8-((5-(((2R,11aS)-2-((tert-Butyldimethylsilyl)oxy)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (58 mg, 73.60 μmol) was dissolved in tetrahydrofuran (1 mL). Tetrabutylammonium fluoride solution (1 M, 145.17 μL) was added dropwise and stirred for 1 hour. The mixture was diluted with ethyl acetate, washed twice with saturated ammonium chloride solution and twice with saturated brine, and concentrated under reduced pressure. The title compound (17.11 mg, 24.24 μmol) was obtained by purification through preparative HPLC and freeze-drying.

[0758] Its structural characterization data are as follows:

[0759] MS m / z(ESI):685.3[M+H] +

[0760] 1 H NMR (400MHz, DMSO) δ7.45 (s, 1H), 7.42-7.36 (m, 3H), 7.29 (s, 1H), 6.91 (d, J = 8.9Hz, 2H), 6.55 (d, J = 5.7Hz, 1H), 6. 36(d,J=5.9Hz,1H),6.31(s,1H),6.26(s,1H),4.92(d,J=3.2Hz,1H),4.19(s,2H),3.93(dd,J=13.7,6.7Hz,4H),3. 80(d,J=9.1Hz,1H),3.76(s,3H),3.65(d,J=9.5Hz,6H),3.51(dt,J=12.1,9.0Hz,4H),3.28-3.19(m,2H),2.99-2. 93(m,1H),2.79-2.74(m,1H),2.08(dd,J=13.0,6.7Hz,1H),1.80(s,4H),1.73-1.67(m,1H),1.56(d,J=6.4Hz,2H).

[0761] Its preparation method is as follows:

[0762] Chromatographic column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0763] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0764] Preparation Example 25: Preparation of (S)-7-methoxy-8-((5-(((S)-7-methoxy-2-methylene-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-41)

[0765] Step 1: Preparation of (S)-4-methylenepyrrolidine-2-carboxylic acid methyl ester (7-41-2)

[0766] Dissolve (S)-1-(tert-Butoxycarbonyl)-4-methylenepyrrolidine-2-carboxylic acid (941.87 mg, 4.14 mmol) in MeOH (7 mL) and add HCl / dioxane (4 M, 7 mL) dropwise. Stir at room temperature for 24 hours. After completion, the reaction mixture was concentrated under reduced pressure to obtain the crude hydrochloride salt of the title compound (794 mg, 4.02 mmol), which was used directly in the next step without purification.

[0767] Its structural characterization data are as follows:

[0768] ESI-MS (m / z): 142.2 [M+H] +

[0769] Step 2: Preparation of (S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-methylenepyrrolidine-2-carboxylic acid methyl ester (7-41-3)

[0770] Dissolve 4-(Benzyloxy)-5-methoxy-2-nitrobenzoic acid (1.35 g, 4.45 mmol) in DMF (15 mL). Add HATU (2.03 g, 5.34 mmol) and DIPEA (1.72 g, 13.34 mmol, 2.32 mL) and stir at room temperature for 10 minutes. Then, add a mixture of (S)-methyl 4-methylenepyrrolidine-2-carboxylate hydrochloride (790 mg, 4.45 mmol) and DIPEA (574.80 mg, 4.45 mmol) in DMF (15 mL). Stir at room temperature for 2 hours. After completion, add 100 mL of water and extract with 50 mL x 3 of ethyl acetate. The organic phase is washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product is purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0-60%) and concentrated again under reduced pressure to obtain the title compound (1.53 g, 3.58 mmol).

[0771] Its structural characterization data are as follows:

[0772] ESI-MS (m / z): 427.4 [M+H] +

[0773] Step 3: Preparation of (S)-8-(benzyloxy)-7-methoxy-2-methylene-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-41-4)

[0774] Methyl (S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-4-methylenepyrrolidine-2-carboxylate (1.53 g, 3.59 mmol) was dissolved in saturated aqueous ammonium chloride (8 mL) and methanol (80 mL). Zinc powder (2.35 g, 35.88 mmol) was added and the mixture was heated to 75°C with stirring for 4 hours. After completion of the reaction, the reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was then extracted with 50 mL of water and 3 50 mL of a 4:1 mixture of ethyl acetate and methanol. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude title compound (1.3 g, 3.57 mmol), which was used directly in the next step without purification.

[0775] Its structural characterization data are as follows:

[0776] ESI-MS (m / z): 365.2 [M+H] +

[0777] Step 4: Preparation of (S)-8-(benzyloxy)-7-methoxy-2-methylene-10-((2-(trimethylsilyl)ethoxy)methyl)-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-41-5)

[0778] Dissolve (S)-8-(Benzyloxy)-7-methoxy-2-methylene-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (0.2 g, 548.86 μmol) in THF (3 mL). Cool to -40°C under nitrogen, then add n-BuLi (2.5 M, 230.52 μL) dropwise. Incubate for 30 minutes. Then, add 2-(Trimethylsilyl)ethoxymethyl chloride (100.66 mg, 603.74 μmol) dropwise at -40°C. After addition, allow to react overnight. After completion of the reaction, the reaction solution was quenched by adding 5 mL of aqueous ammonium chloride solution, and then extracted with 10 mL of water and 10 mL of ethyl acetate × 3. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain a crude product of the title compound (286 mg, 520.36 μmol), which was used directly in the next reaction without purification.

[0779] Its structural characterization data are as follows:

[0780] ESI-MS (m / z): 496.3 [M+H] +

[0781] Step 5: Preparation of (S)-8-(benzyloxy)-7-methoxy-2-methylene-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-41-6)

[0782] (S)-8-(Benzyloxy)-7-methoxy-2-methylene-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (180 mg, 363.89 μmol) was dissolved in THF (10 mL) and cooled to -78°C under nitrogen. Lithium triethylborohydride (1 M, 727.78 μL) was then added dropwise. The mixture was incubated for 2 hours. Additional lithium triethylborohydride (1 M, 727.78 μL) was added three times, with a one-hour interval between each addition. After completion of the reaction, the reaction mixture was added to 15 mL of water and extracted with ethyl acetate (3 times, 10 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford the crude title compound (200 mg, 344.44 μmol), which was used directly in the next step without purification.

[0783] Its structural characterization data are as follows:

[0784] ESI-MS (m / z): 367.3 [M+H+H2O] + ; 697.4[2M+H] +

[0785] Step 6: Preparation of (S)-8-(benzyloxy)-7-methoxy-2-methylene-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-41-7)

[0786] (S)-8-(Benzyloxy)-7-methoxy-2-methylene-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (80 mg, 229.62 μmol) was dissolved in dichloromethane (1 mL) and methanol (1 mL). Sodium triacetoxyborohydride (97.33 mg, 459.25 μmol) was added and stirred at room temperature overnight for 16 hours. NaBH4 (17.37 mg, 459.25 μmol) was added and allowed to react for 2 hours. Additional NaBH4 (17.37 mg, 459.25 μmol) was added and allowed to react for another 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, dissolved in methanol, purified by reverse-phase column chromatography (acetonitrile / 0.05% formic acid aqueous solution = 0-50%), and freeze-dried to obtain the title compound (15 mg, 42.81 μmol).

[0787] Its structural characterization data are as follows:

[0788] ESI-MS (m / z): 351.3 [M+H] +

[0789] Step 7: Preparation of (S)-8-hydroxy-7-methoxy-2-methylene-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-41-8)

[0790] To (S)-8-(Benzyloxy)-7-methoxy-2-methylene-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (15 mg, 42.81 μmol) was added dichloromethane (1 mL), followed by the dropwise addition of methanesulfonic acid (0.1 mL). The mixture was stirred at room temperature for 1 hour. After completion of the reaction, 2 mL of water was added, and the pH was adjusted to 8-9 with saturated sodium bicarbonate solution. The mixture was extracted with 3 mL of dichloromethane x 3. The organic phase was washed with 3 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford the crude title compound (11 mg, 42.26 μmol), which was used directly in the next step without purification.

[0791] Its structural characterization data are as follows:

[0792] ESI-MS (m / z): 261.1 [M+H] +

[0793] Step 8: Preparation of (S)-7-methoxy-8-((5-(((S)-7-methoxy-2-methylene-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo)[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (7-41-9)

[0794] (S)-8-((5-bromopentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (24.74 mg, 42.26 μmol) and (S)-8-hydroxy-7-methoxy-2-methylene-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5-one (11 mg, 42.26 μmol) were dissolved in DMF (1 mL). Potassium carbonate (17.52 mg, 126.78 μmol) was added and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction solution was directly purified by preparative HPLC and freeze-dried to obtain the title compound (5 mg, 6.54 μmol).

[0795] Its structural characterization data are as follows:

[0796] ESI-MS (m / z): 766.3 [M+H] +

[0797] The preparation method is as follows:

[0798] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0799] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0800] Step 9: Preparation of (S)-7-methoxy-8-((5-(((S)-7-methoxy-2-methylene-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-41)

[0801] (S)-7-Methoxy-8-((5-(((S)-7-methoxy-2-methylene-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo)[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (5 mg, 6.54 μmol) was dissolved in DMF (0.5 mL), tetrahydropyrrole (464.92 μg, 6.54 μmol) and tetrakistriphenylphosphine palladium (755.41 μg, 6.54e-1 μmol) were added, and the nitrogen atmosphere was replaced three times. The reaction was then stirred at room temperature under nitrogen protection for 2 hours. After the reaction was completed, the reaction solution was directly purified by preparative HPLC and freeze-dried to obtain the title compound (1.89 mg, 2.47 μmol).

[0802] Its structural characterization data are as follows:

[0803] ESI-MS (m / z): 681.4 [M+H] +

[0804] The preparation method is as follows:

[0805] Chromatographic column: Waters SunFire Prep C18 OBD 19mm×150mm×5.0μm

[0806] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0807] Preparation Example 26: Preparation of (1aR,9aS,10aR)-5-methoxy-6-((5-(((S)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1a,8,9,9a,10,10a-hexahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepin-3(1H)-one (7-43)

[0808] Step 1: Preparation of (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid methyl ester hydrochloride (7-43-2)

[0809] Dissolve (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid (2.00 g, 8.80 mmol) in methanol (10 mL), add 4 M hydrochloric acid in dioxane (10 mL) dropwise, and stir for 4 hours. After concentration under reduced pressure, proceed directly to the next step.

[0810] Step 2: Preparation of (1R,3S,5R)-2-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid methyl ester (7-43-3)

[0811] Dissolve 4-benzyloxy-5-methoxy-2-nitro-benzoic acid (1.18 g, 3.89 mmol), (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid methyl ester hydrochloride (760.26 mg, 4.28 mmol), and HATU (2.22 g, 5.84 mmol) in DMF (20 mL). Add DIPEA (1.51 g, 11.67 mmol) dropwise and stir for 2 hours. Add water and EA, stir, and separate. The aqueous phase is extracted twice with EA. The combined organic phases are washed twice with saturated brine, dried, and concentrated. Purify on a silica gel column (eluent: 30-65% ethyl acetate / petroleum ether) and reconcentrate to obtain the title compound (1.64 g, 3.85 mmol).

[0812] Step 3: Preparation of (1aR,9aS,10aR)-6-(benzyloxy)-5-methoxy-1a,9a,10,10a-tetrahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepine-3,9(1H,8H)-dione (7-43-4)

[0813] Dissolve methyl (1R,3S,5R)-2-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-2-azabicyclo[3.1.0]hexane-3-carboxylate (870 mg, 2.04 mmol) in methanol (10 mL). Add saturated aqueous ammonium chloride (2 mL). Add zinc powder (1.33 g, 20.40 mmol) portionwise. Heat to 80°C and reflux for 16 hours. Cool to room temperature, filter, and concentrate the filtrate under reduced pressure. Purify on a silica gel column (eluent: 5% methanol / dichloromethane) and reconcentrate to obtain the title compound (682 mg, 1.87 mmol).

[0814] Its structural characterization data are as follows:

[0815] MS m / z(ESI):365.1[M+H] +

[0816] Step 4: Preparation of (1aR,9aS,10aR)-6-(benzyloxy)-5-methoxy-1a,9a,10,10a-tetrahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepine-3,9(1H,8H)-dione (7-43-5)

[0817] Dissolve (1aR,9aS,10aR)-6-(benzyloxy)-5-methoxy-1a,9a,10,10a-tetrahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepine-3,9(1H,8H)-dione (180 mg, 493.97 μmol) in dry THF (10 mL). Add sodium borohydride (56.06 mg, 1.48 mmol) with stirring, and then add a solution of trifluoroacetic acid (112.65 mg, 987.94 μmol, 75.65 μL) in tetrahydrofuran (100 μL) dropwise, producing abundant bubbling. Heat to 75°C and stir for 4 hours. The mixture was concentrated under reduced pressure, and water and ethyl acetate were added and stirred. The layers were separated, and the organic phase was concentrated under reduced pressure. Purification was performed on a silica gel column (eluent: 5% methanol / dichloromethane) and concentrated again to obtain the title compound (142 mg, 405.24 μmol).

[0818] Its structural characterization data are as follows:

[0819] MS m / z(ESI):367.3[M+H2O] +

[0820] Step 5: Preparation of (1aR,9aS,10aR)-6-(benzyloxy)-5-methoxy-3-oxo-1,1a,9,9a,10,10a-hexahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepine-8(3H)-carboxylic acid allyl ester (7-43-6)

[0821] Dissolve (1aR,9aS,10aR)-6-(benzyloxy)-5-methoxy-1a,9a,10,10a-tetrahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepine-3,9(1H,8H)-dione (142 mg, 405.24 μmol) in dry dichloromethane (5 mL). Add pyridine (48.08 mg, 607.86 μmol) dropwise. Cool to 0°C with stirring. Add allyl chlorate (195.38 mg, 1.62 mmol, 172.29 μL) dropwise. Return to room temperature and stir for 1 hour. Dilute with dichloromethane, add 10% citric acid solution, stir, and allow to stand for separation. Wash the organic phase with water, dry, and concentrate. The residue was purified by silica gel column (eluent: 80% ethyl acetate / petroleum ether) and concentrated again to obtain the title compound (153 mg, 352.14 μmol).

[0822] Step 6: Preparation of (1aR,9aS,10aR)-6-hydroxy-5-methoxy-3-oxo-1,1a,9,9a,10,10a-hexahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepine-8(3H)-carboxylic acid allyl ester (7-43-7)

[0823] Dissolve allyl (1aR,9aS,10aR)-6-(benzyloxy)-5-methoxy-3-oxo-1,1a,9,9a,10,10a-hexahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepine-8(3H)-carboxylate (130 mg, 299.21 μmol) in dichloromethane (1 mL). Add methanesulfonic acid (0.5 mL) dropwise with stirring and continue the reaction for 1 hour. Dilute with dichloromethane, then add aqueous sodium bicarbonate solution and stir. Allow to stand for separation, and the organic phase is dried and concentrated. Purify by preparative HPLC and freeze-dry to afford the title compound (57 mg, 165.52 μmol).

[0824] Its structural characterization data are as follows:

[0825] MS m / z(ESI):345.1[M+H] +

[0826] Its preparation method is as follows:

[0827] Chromatographic column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0828] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0829] Step 7: Preparation of (1aR,9aS,10aR)-6-((5-(((S)-10-((allyloxy)carbonyl)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-5-methoxy-3-oxo-1,1a,9,9a,10,10a-hexahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepin-8(3H)-carboxylic acid allyl ester (7-43-8)

[0830] (S)-8-((5-bromopentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (10.20 mg, 17.42 μmol), (1aR,9aS,10aR)-6-hydroxy-5-methoxy-3-oxo-1,1a,9,9a,10,10a-hexahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepine-8(3H)-carboxylic acid allyl ester (5 mg, 14.52 μmol), and potassium carbonate (3.01 mg, 21.78 μmol) were dissolved in DMF (1 mL) and stirred for 16 hours. Water was added to precipitate a solid, which was filtered, washed with water, and dried in vacuo to give the title compound (12 mg, 14.14 μmol).

[0831] Its structural characterization data are as follows:

[0832] MS m / z(ESI):849.4[M+H] +

[0833] Step 8: Preparation of (1aR,9aS,10aR)-5-methoxy-6-((5-(((S)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-1a,8,9,9a,10,10a-hexahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepin-3(1H)-one (7-43)

[0834] (1aR,9aS,10aR)-6-((5-(((S)-10-((allyloxy)carbonyl)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-5-methoxy-3-oxo-1,1a,9,9a,10 1,0a-Hexahydrobenzo[e]cyclopropyl[4,5]pyrrolo[1,2-a][1,4]diazepine-8(3H)-carboxylic acid allyl ester (12 mg, 14.14 μmol) was dissolved in DMF (1 mL). Tetrahydropyrrole (1.01 mg, 14.14 μmol) was added dropwise. Tetrakis(triphenylphosphine)palladium (1.63 mg, 1.41 μmol) was then added. The reaction was continued under nitrogen atmosphere for 16 hours. The title compound (6.18 mg, 8.99 μmol) was obtained by purification by preparative HPLC and freeze-drying.

[0835] Its structural characterization data are as follows:

[0836] MS m / z(ESI):681.3[M+H] +

[0837] Its preparation method is as follows:

[0838] Chromatographic column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0839] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0840] Preparation Example 27: Preparation of (S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-44)

[0841] Step 1: Preparation of (S)-8-(benzyloxy)-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (7-44-1)

[0842] Dissolve (4-(Benzyloxy)-5-methoxy-2-nitrobenzoyl)-L-proline methyl ester (870 mg, 2.04 mmol) in methanol (100 mL). Add saturated aqueous ammonium chloride (10 mL). Add zinc powder (2.84 g, 43.44 mmol) portionwise. Heat to 80°C and reflux for 16 hours. Cool to room temperature, filter, and concentrate the filtrate under reduced pressure. Purify on a silica gel column (eluent: 5% methanol / dichloromethane) and reconcentrate to obtain the title compound (1.15 g, 3.26 mmol).

[0843] Its structural characterization data are as follows:

[0844] MS m / z(ESI):353.1[M+H] +

[0845] Step 2: Preparation of (S)-8-(benzyloxy)-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-44-2)

[0846] Dissolve (S)-8-(Benzyloxy)-7-methoxy-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dione (1.1 g, 3.12 mmol) in dry THF (10 mL). Add sodium borohydride (354.29 mg, 9.36 mmol) with stirring, followed by a solution of trifluoroacetic acid (711.85 mg, 6.24 mmol, 478.07 μL) in tetrahydrofuran (100 μL) dropwise, resulting in abundant bubbling. Heat to 75°C and stir for 4 hours. Concentrate under reduced pressure, add water and ethyl acetate, stir, separate, and concentrate the organic phase under reduced pressure. Purify on a silica gel column (eluent: 5% methanol / dichloromethane), and reconcentrate to obtain the title compound (1.01 g, 2.98 mmol).

[0847] Step 3: Preparation of (S)-8-(benzyloxy)-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (7-44-3)

[0848] Dissolve (S)-8-(Benzyloxy)-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (1 g, 2.96 mmol) in dry dichloromethane (50 mL). Add pyridine (350.62 mg, 4.43 mmol, 358.51 μL) dropwise. Cool to 0°C with stirring. Add allyl chlorate (1.07 g, 8.87 mmol, 942.30 μL) dropwise. Return to room temperature and stir for 1 hour. Dilute with dichloromethane, add 10% citric acid solution, stir, and allow to stand for separation. Wash the organic phase with water, dry, and concentrate. Purify on a silica gel column (eluent: 80% ethyl acetate / petroleum ether) and reconcentrate to obtain the title compound (530 mg, 1.25 mmol).

[0849] Step 4: Preparation of (S)-8-hydroxy-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (7-44-4)

[0850] Dissolve (S)-8-(Benzyloxy)-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (530 mg, 1.25 mmol) in dichloromethane (4 mL). Add methanesulfonic acid (2 mL) dropwise with stirring and continue the reaction for 1 hour. Dilute with dichloromethane, then add aqueous sodium bicarbonate solution with stirring. Allow to stand for separation, and the organic phase is dried and concentrated to give the title compound (410 mg, 1.23 mmol).

[0851] Its structural characterization data are as follows:

[0852] MS m / z(ESI):333.2[M+H] +

[0853] Step 5: Preparation of (S)-8-((5-(((S)-10-((allyloxy)carbonyl)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (7-44-5)

[0854] (S)-8-((5-bromopentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (40 mg, 68.32 μmol), (S)-8-hydroxy-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (22.71 mg, 68.32 μmol), and potassium carbonate (14.16 mg, 102.48 μmol) were dissolved in DMF (1 mL) and stirred for 16 hours. Water was added to precipitate a solid, which was filtered, washed with water, and dried in vacuo to give the title compound (55 mg, 65.72 μmol).

[0855] Its structural characterization data are as follows:

[0856] MS m / z(ESI):838.4[M+H] +

[0857] Step 6: Preparation of (S)-7-methoxy-8-((5-(((S)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-2-(4-methoxyphenyl)-1,10,11,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one (7-44)

[0858] (S)-8-((5-(((S)-10-((allyloxy)carbonyl)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-10(5H)-carboxylic acid allyl ester (55 mg, 65.72 μmol) was dissolved in DMF (1 mL), tetrahydropyrrole (4.67 mg, 65.72 μmol) was added dropwise, and tetrakistriphenylphosphine palladium (7.59 mg, 6.57 μmol) was added. The reaction was carried out under nitrogen replacement and protection for 16 hours. Purify by preparative HPLC and freeze-dry to give the title compound (11.71 mg, 17.33 μmol).

[0859] Its structural characterization data are as follows:

[0860] MS m / z(ESI):669.3[M+H] +

[0861] 1 H NMR (400MHz, CDCl3) δ7.49(s,2H),7.31(d,J=8.7Hz,2H),7.26(s,3H),6.89(d,J=8.8Hz,2H),4.29(s,1H),3.99(s,4H),3.87(s,5H),3.81(d,J=11 .6Hz,5H),3.70(s,1H),3.58(d,J=10.2Hz,4H),3.32(s,3H),2.74(dd,J= 16.1,3.6Hz,1H),2.21(dd,J=12.8,6.3Hz,1H),1.90(m,6H),1.65(m,4H).

[0862] Its preparation method is as follows:

[0863] Chromatographic column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0864] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0865] Preparation Example 28: Preparation of (11aS,11a'S)-8,8'-(pentane-1,5-diylbis(oxy))bis(7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one) (7-45)

[0866] Step 1: Preparation of (S)-8-((5-(((R)-10-((allyloxy)carbonyl)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylic acid allyl ester (7-45-1)

[0867] Dissolve 1,5-dibromopentane (12.45 mg, 54.16 μmol), (S)-8-hydroxy-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate (40 mg, 120.35 μmol), and potassium carbonate (49.90 mg, 361.06 μmol) in DMF (1 mL) and stir for 16 hours. Add water to precipitate a solid, which is filtered, washed with water, and dried in vacuo to yield the title compound (40 mg, 54.58 μmol).

[0868] Its structural characterization data are as follows:

[0869] MS m / z(ESI):733.4[M+H] +

[0870] Step 2: Preparation of (11aS, 11a'S)-8,8'-(pentane-1,5-diylbis(oxy))bis(7-methoxy-1,2,3,10,11,11a-hexahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-5-one) (7-45)

[0871] (S)-8-((5-(((R)-10-((allyloxy)carbonyl)-7-methoxy-5-oxo-2,3,5,10,11,11a-hexahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yl)oxy)pentyl)oxy)-7-methoxy-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-10(5H)-carboxylic acid allyl ester (40 mg, 54.58 μmol) was dissolved in DMF (1 mL), tetrahydropyrrole (3.88 mg, 54.58 μmol) was added dropwise, and tetrakis(triphenylphosphine)palladium (6.31 mg, 5.46 μmol) was added. The reaction was carried out under nitrogen replacement and protection for 16 hours. The residue was purified by preparative HPLC and freeze-dried to give the title compound (4.96 mg, 8.70 μmol).

[0872] Its structural characterization data are as follows:

[0873] MS m / z(ESI):565.3[M+H] +

[0874] 1 H NMR (400MHz, CDCl3) δ7.52(s,2H),6.13(s,2H),3.99(s,4H),3.95-3.73(m,9H),3.70(s,2H),3.57(d,J=11 .7Hz,2H),3.29(s,3H),2.22(dd,J=12.6,6.1Hz,2H),1.90(dt,J=13.4,6.5Hz,8H),1.68(d,J=20.9Hz,6H).

[0875] Its preparation method is as follows:

[0876] Chromatographic column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0877] Mobile phase A: acetonitrile; Mobile phase B: water (0.05% formic acid)

[0878] Test example: Inhibitory effect of compounds on tumor cell proliferation

[0879] 1. Inhibitory effect of the compound on NCI-N87 cell proliferation

[0880] (1) Cell plating: First, culture the tumor cells NCI-N87 in the corresponding culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to the appropriate concentration for plating. The tumor cell sources are shown in Table 1.

[0881] Table 1. Tumor cell sources

[0882] Co-incubation of the compound of the present invention and tumor cells: After the cells adhere to the wall, the culture medium in the cells is removed, and the diluted bioactive molecule (the compound of the present invention) is added to the above plate wells and incubated for 72 hours.

[0883] In vitro cell viability assay: After incubation, add 50 μL of Cell Counting-Lite™ 2.0 reagent (Vazyme / Novozyme) to each well, shake and mix thoroughly in the dark. After 10 minutes of reaction, assay can be performed using a microplate reader (Manufacturer: BMG, Model: PHERAStar-FS). Background RLU values ​​are calculated using Cell Counting-Lite™ reagents for cell-free medium, and vehicle RLU values ​​are calculated using Cell Counting-Lite™ reagents for cell-containing medium. Cell inhibition rate = 1 - (sample RLU - background RLU) / (vehicle RLU - background RLU) × 100%. The half-maximal inhibitory concentration (IC50) of the compound was calculated using a four-parameter curve fit. RLU (relative light unit): relative light unit. The assay results are shown in Tables 2-1 and 2-2.

[0884] (2) Data results

[0885] Table 2-1. NCI-N87 cell proliferation inhibitory activity

[0886] Table 2-2. NCI-N87 cell proliferation inhibitory activity

[0887] The test results show that the compound of the present invention has a significant inhibitory effect on the proliferation of NCI-N87 human gastric cancer cells.

[0888] 2. Inhibitory effect of compounds on HT29 cell proliferation

[0889] (1) Cell plating: First, culture HT29 tumor cells in the corresponding culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to an appropriate concentration for plating. The sources of tumor cells are shown in Table 3.

[0890] Table 3. Tumor cell origin

[0891] Co-incubation of the compound of the present invention and tumor cells: After the cells adhere to the wall, the culture medium in the cells is removed, and the diluted bioactive molecule (the compound of the present invention) is added to the above plate wells and incubated for 72 hours.

[0892] In vitro cell viability assay: After incubation, add 50 μL of Cell Counting-Lite™ 2.0 reagent (Vazyme / Novozyme) to each well, shake and mix in the dark. After 10 minutes of reaction, assay can be performed using a microplate reader (Manufacturer: BMG, Model: PHERAStar-FS). Background RLU values ​​are calculated using Cell Counting-Lite™ culture medium without cells, and vehicle RLU values ​​are calculated using Cell Counting-Lite™ culture medium with cells. Cell inhibition rate = 1 - (sample RLU - background RLU) / (vehicle RLU - background RLU) × 100%. The half-maximal inhibitory concentration (IC50) of the compound was calculated using a four-parameter curve fit. RLU (relative light unit): relative light unit. The assay results are shown in Tables 4-1 and 4-2.

[0893] (2) Data results

[0894] Table 4-1. HT29 cell proliferation inhibitory activity

[0895] Table 4-2. HT29 cell proliferation inhibitory activity

[0896] The test results show that the compound of the present invention has a strong proliferation inhibitory effect on HT29 colon cancer cell proliferation.

[0897] 3. Inhibitory effect of compounds on HCC827 cell proliferation

[0898] (1) Cell plating: First, culture HCC827 tumor cells in the appropriate culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to an appropriate concentration for plating. The sources of tumor cells are shown in Table 5.

[0899] Table 5. Tumor cell origin

[0900] Co-incubation of the compound of the present invention and tumor cells: After the cells adhere to the wall, the culture medium in the cells is removed, and the diluted bioactive molecule (the compound of the present invention) is added to the above plate wells and incubated for 72 hours.

[0901] In vitro cell viability assay: After incubation, add 50 μL of Cell Counting-Lite™ 2.0 reagent (Vazyme / Novozyme) to each well, shake and mix in the dark. After 10 minutes of reaction, assays can be performed using a microplate reader (Manufacturer: BMG, Model: PHERAStar-FS). Background RLUs were calculated using Cell Counting-Lite™ reagents for cell-free medium, and vehicle RLUs were calculated using Cell Counting-Lite™ reagents for cell-containing medium. Cell inhibition rate = 1 - (sample RLU - background RLU) / (vehicle RLU - background RLU) × 100%. The half-maximal inhibitory concentration (IC50) of the compound was calculated using a four-parameter curve fit. RLU (relative light unit): relative light unit. The assay results are shown in Table 6.

[0902] (2) Data results

[0903] Table 6. Antiproliferation activity of compounds on HCC827 cells

[0904] The test results show that the compound of the present invention has an inhibitory effect on the proliferation of HCC827 human non-small cell lung cancer cells.

[0905] 4. Inhibitory effect of the compounds on HCC1954 cell proliferation

[0906] (1) Cell plating: First, culture HCC1954 tumor cells in the corresponding culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to an appropriate concentration for plating. The tumor cell sources are shown in Table 7.

[0907] Table 7. Tumor cell sources

[0908] Co-incubation of the compound of the present invention and tumor cells: After the cells adhere to the wall, the culture medium in the cells is removed, and the diluted bioactive molecule (the compound of the present invention) is added to the above plate wells and incubated for 72 hours.

[0909] In vitro cell viability assay: After incubation, add 50 μL of Cell Counting-Lite™ 2.0 reagent (Vazyme / Novozyme) to each well, shake and mix in the dark. After 10 minutes of reaction, assay can be performed using a microplate reader (Manufacturer: BMG, Model: PHERAStar-FS). Background RLU values ​​are calculated using Cell Counting-Lite™ for cell-free culture medium, and vehicle RLU values ​​are calculated using Cell Counting-Lite™ for cell-containing culture medium. Cell inhibition rate = 1 - (sample RLU - background RLU) / (vehicle RLU - background RLU) × 100%. The half-maximal inhibitory concentration (IC50) of the compound was calculated using a four-parameter curve fit. RLU (relative light unit): relative light unit. The assay results are shown in Tables 8-1 and 8-2.

[0910] (2) Data results

[0911] Table 8-1. Antiproliferation activity of compounds on HCC1954 cells

[0912] Table 8-2. Proliferation inhibition activity of compounds on HCC1954 cells

[0913] The test results show that the compound of the present invention has an inhibitory effect on the proliferation of HCC1954 human non-small cell lung cancer cells.

[0914] 5. Inhibitory effect of compounds on NCI-H1975 cell proliferation

[0915] (1) Cell plating: First, culture NCI-H1975 tumor cells in the corresponding culture medium, digest the cells with trypsin, centrifuge, resuspend the cells, count the cells, and adjust the cells to an appropriate concentration for plating. The source of tumor cells is shown in Table 9.

[0916] Table 9. Tumor cell sources

[0917] Co-incubation of the compound of the present invention and tumor cells: After the cells adhere to the wall, the culture medium in the cells is removed, and the diluted bioactive molecule (the compound of the present invention) is added to the above plate wells and incubated for 72 hours.

[0918] In vitro cell viability assay: After incubation, add 50 μL of Cell Counting-Lite™ 2.0 reagent (Vazyme / Novozyme) to each well, shake and mix in the dark. After 10 minutes of reaction, assays can be performed using a microplate reader (Manufacturer: BMG, Model: PHERAStar-FS). Background RLUs were calculated using Cell Counting-Lite™ reagents for cell-free medium, and vehicle RLUs were calculated using Cell Counting-Lite™ reagents for cell-containing medium. Cell inhibition rate = 1 - (sample RLU - background RLU) / (vehicle RLU - background RLU) × 100%. The half-maximal inhibitory concentration (IC50) of the compound was calculated using a four-parameter curve fit. RLU (relative light unit): relative light unit. The assay results are shown in Table 10.

[0919] (2) Data results

[0920] Table 10. Antiproliferation activity of compounds on NCI-H1975 cells

[0921] The test results show that the compound of the present invention has an inhibitory effect on the proliferation of NCI-H1975 human non-small cell lung cancer cells.

[0922] Although the specific embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that various modifications and substitutions may be made to those details based on all the teachings disclosed, and these changes are all within the scope of protection of the present invention. The full scope of the present invention is given by the appended claims and any equivalents thereof.

Claims

1. A compound or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein the compound has the structure shown below: in, R 1 and R 2 are each independently selected from hydrogen, -CN, halogen, -OR a 、-NH2、-NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally substituted by one or more selected from -CN, halogen, -OH, -NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 substituted by aryl and 5-10 membered heteroaryl substituents; R 4 and R 4’ are each independently selected from hydrogen, halogen, -OH, -NH2, C 1-6 Alkyl, halogenated C 1-6 Alkyl and C 1-6 Alkoxy, R 5 and R 5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms of the 5 and R 5’ are all hydrogen; or, R 4 and R 5 Connected to form a bond or together with R 4 and R 5 The carbon atoms form a 3-6-membered carbon ring, R 5’ and R 4’ are each independently selected from hydrogen, halogen, -NH2, C 1-6 Alkyl, halogenated C 1-6 Alkyl and C 1-6 Alkoxy; R 6 For hydrogen, C 1-6 Alkyl or R 6’ , R 7 and R 7’ are all hydrogen; or, R 6’ With R 7 Connected into a bond, R 7’ is hydrogen; R a Selected from H, C 1-6 Alkyl or C 3-6 Cycloalkyl; Ring A is selected from 3-6 membered cycloalkyl, C 6-10 Aryl, 5-10 membered heteroaryl, 3-6 membered heterocyclic group; R 3 Selected from hydrogen, -CN, halogen, -OR a 、-NH2、-NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally substituted by one or more selected from -CN, halogen, -OH, -NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 substituted by aryl and 5-10 membered heteroaryl substituents; X and Y are each independently -CH2-, -CD2-, and -C(O)-; m is selected from 1-5, for example 1, 2, 3, 4 or 5; n is selected from 1-10, for example 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; When n is 5 and ring A is phenyl, R 3 Not methoxy.

2. The compound of claim 1 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein R 1 Selected from hydrogen, -CN, halogen, -OR a and C 1-6 Alkyl, where R a Selected from C 1-6 Alkyl, deuterated C 1-6 Alkyl and C 3-6 Cycloalkyl; Preferably, R 1 Select from -OR a , where R a Selected from C 1-6 Alkyl, deuterated C 1-6 Alkyl and C 3-6 Cycloalkyl; Preferably, R 1 is selected from methoxy, trideuterated methoxy and -O-cyclopropyl.

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein R 2 Selected from hydrogen, -CN, halogen, -OR a and C 1-6 Alkyl, where R a C 1-6 Alkyl or C 3-6 Cycloalkyl; Preferably, R2 For-OR a , where R a C 1-6 alkyl; Preferably, R 2 Selected from methoxy.

4. The compound of any one of claims 1 to 3 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein R 4 is hydrogen or halogen, R 4’ Selected from hydrogen, halogen, -OH, -NH2, C 1-6 Alkyl, halogenated C 1-6 Alkyl and C 1-6 Alkoxy, R 5 and R 5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms of the 5 and R 5’ are all hydrogen; or, R 4 and R 5 Connected to form a bond or together with R 4 and R 5 The carbon atoms form a 3-6-membered carbon ring, R 5’ and R 4’ All are hydrogen; Preferably, R 4 is hydrogen or fluorine, R 4’ is selected from hydrogen, fluorine, -OH, -NH2, methyl, trifluoromethyl and methoxy, R 5 and R 5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms of the 5 and R 5’ are all hydrogen; or, R 4 and R 5 Connected to form a bond or together with R 4 and R 5 The carbon atoms form a 3-membered carbon ring, R 5’ and R 4’ All are hydrogen; Preferably, R 4 is hydrogen or fluorine, R 4’ is selected from hydrogen, fluorine, -OH, -NH2, methyl, trifluoromethyl and methoxy, R 5 and R5’ are all hydrogen; or, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms form a carbon-carbon double bond, R 5 and R 5’ are all hydrogen; or, R 4 and R 5 Connected to form a bond or together with R 4 and R 5 The carbon atoms form a 3-membered carbon ring, R 5’ and R 4’ All are hydrogen; Preferably, R 4 and R 4’ Connect with R 4 and R 4’ The carbon atoms form a 3-membered carbon ring, R 5 and R 5’ All are hydrogen.

5. The compound of any one of claims 1 to 4 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein R 6 for Ring A is selected from 3-6 membered cycloalkyl, C 6-10 Aryl or 5-10 membered heteroaryl; R 3 Selected from hydrogen, -CN, halogen, -OR a 、-NH2、-NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl; the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally substituted by one or more selected from -CN, halogen, -OH, -NH2, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, C 6-10 substituted by aryl and 5-10 membered heteroaryl substituents; R a Selected from H and C 1-6 alkyl; m is selected from 1, 2, 3, 4 and 5; Preferably, R 6 for Ring A is selected from cyclopropyl, phenyl, furanyl, thienyl, pyrimidinyl and pyridinyl; R 3 is selected from hydrogen, fluorine, -OH, methoxy, -NH2 and hydroxymethyl; m is 1 or 2; Preferably, R 6 for Ring A is phenyl; R 3 is selected from hydrogen, fluorine, -OH, methoxy, -NH2 and hydroxymethyl; m is 1 or 2.

6. The compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein X is -CH2- or -CD2-; optionally, Y is selected from -CH2-, -CD2-, and -C(O)-; Preferably, X is -CH2-, and Y is -CH2-; X is -CH2-, and Y is -CD2- or -C(O)-; or X is -CD2-, and Y is -CH2-; Preferably, X is -CH2- and Y is -CH2-.

7. The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein the compound has a structure as shown in the following formula (I-1) or formula (I-2): in, R 1 , R 2 , R 3 , R 4 , R 4’ , R 5 , R 5’ , Ring A, m and n are as described in any one of claims 1-6.

8. The compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein the compound has a structure as shown in the following formula (II-1) or formula (II-2): in, R 1 , R 2 , R 3 , R 4 , R 4’ , R 5 , R 5’ , Ring A, m and n are as described in any one of claims 1-6.

9. The compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, wherein the compound is selected from the following structures:

10. An antibody drug conjugate comprising a compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled product, metabolite or prodrug fragment thereof.

11. A pharmaceutical composition comprising the compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, or the antibody-drug conjugate of claim 10, and one or more pharmaceutical excipients.

12. Use of the compound of any one of claims 1 to 9 or its pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug, the antibody drug conjugate of claim 10, or the pharmaceutical composition of claim 11 in the preparation of a drug for treating cancer or tumor; Optionally, the cancer or tumor is a solid tumor or a hematological malignancy; for example selected from gastric cancer, breast cancer, lung cancer (eg, non-small cell lung cancer, in particular lung adenocarcinoma) and urothelial carcinoma.

13. The compound of any one of claims 1 to 9 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope-labeled substance, metabolite or prodrug thereof, the antibody drug conjugate of claim 10, or the pharmaceutical composition of claim 11, for use in treating cancer or tumors; Optionally, the cancer or tumor is a solid tumor or a hematological malignancy; for example selected from gastric cancer, breast cancer, lung cancer (eg, non-small cell lung cancer, in particular lung adenocarcinoma) and urothelial carcinoma.

14. A method for treating cancer or tumors, comprising administering a therapeutically effective amount of a compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, nitrogen oxide, isotope label, metabolite or prodrug thereof, an antibody drug conjugate according to claim 10, or a pharmaceutical composition according to claim 11 to a subject in need thereof; optionally, the cancer or tumor is a solid tumor or a hematological malignancy; for example, selected from gastric cancer, breast cancer, lung cancer (e.g., non-small cell lung cancer, specifically lung adenocarcinoma) and urothelial carcinoma.