Targeted pyrrolobenzodiazapine conjugate

Abstract

Compounds comprising pyrrolobenzodiazepine (PBD) conjugates and methods for using such conjugates are provided. In some embodiments, the conjugate is of formula (I), or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, where each of rings A and B is independently one of the following formulas, namely formulas (IIa), (IIb), (IIc), (IId), (IIe), (IIf), and (IIg), and the remaining variable elements (e.g., linker, ring C, R) 1 , R 2 , R 3 , R 4 , R 5 The values ​​of m, n, and o are as described herein. TIFF2026514000000140.tif154165

Description

[Technical Field]

[0001] 1. Cross-reference of related applications This application claims priority to International Application PCT / CN2023 / 088051, filed on 13 April 2023, the entirety of which is incorporated herein by reference.

[0002] 2. Technical Fields This disclosure relates to targeted pyrrolobenzodiazepine (PBD) conjugates.

[0003] 3. Sequence Listing This application includes a sequence listing submitted electronically in XML format, the entirety of which is incorporated herein by reference. The above XML copy was created on March 26, 2024, is named "01368-0071-00PCT_SL", and has a size of 3,443 bytes. [Background technology]

[0004] 4.Background technology Some pyrrolobenzodiazepines (PBDs) have the ability to recognize and bind to specific sequences of DNA. A preferred sequence is PuGPu. Anthramycin, the first PBD antitumor antibiotic, was discovered in 1965 (Leimgruber, et al., J.Am.Chem.Soc., 87, 5793-5795 (1965); Leimgruber, et al., J.Am.Chem.Soc., 87, 5791-5793 (1965)). Since then, many naturally occurring PBDs have been reported, and more than 10 synthetic pathways have been developed for various analogues (Thurston, et al., Chem.Rev. 1994, 433-465 (1994)).Family members include abeimycin (Hochlowski, et al., J. Antibiotics, 40, 145-148 (1987)), thikamycin (Konishi, et al., J. Antibiotics, 37, 200-206 (1984)), DC-81 (Japanese Patent No. 58-180 487; Thurston, et al., Chem. Brit., 26, 767-772 (1990); Bose, et al., Tetrahedron, 48, 751-758 (1992)), mazetramycin (Kuminoto, et al., J. Antibiotics, 33, 665-667 (1980)), and neotramycin A and B (Takeuchi, et al. al., J. Antibiotics, 29, 93-96 (1976)), Polotoramycin (Tsunakawa, et al., J. Antibiotics, 41, 1366-1373 (1988)), Protracalcin (Shimizu, et al, J. Antibiotics, 29, 2492-2503 (1982); Langley and Thurston, J. Org. Chem., 52, 91-97 (1987)), Cibanomycin (DC-102) (Hara, et al., J. Antibiotics, 41, 702-704 (1988); Itoh, et al., J. Antibiotics, 41, 1281-1284 (1988)), Cibilomycin (Leber, et Examples include al., J. Am. Chem. Soc., 110, 2992-2993 (1988), and tomamycin (Arima, et al., J. Antibiotics, 25, 437-444 (1972)). PBD has the following general structure: [ka]

[0005] PBDs differ in the number, type, and position of substituents on both their aromatic A and pyrrolo C rings, as well as the degree of saturation of the C ring. The B ring contains an imine (N=C), carbinolamine (NH-CH(OH)), or carbinolamine methyl ether (NH-CH(OMe)) at the N10-C11 position, which is the electrophilic center responsible for DNA alkylation. All known natural products have a (S)-configuration at the chiral C11a position, which imparts a clockwise twist to the A ring when viewed from the C ring. This imparts a three-dimensional shape to these DNA molecules that is suitable for isohelicity in relation to the minor groove of type B DNA, resulting in a tight fit at the binding site (Kohn, In Antibiotics III. Springer-Verlag, New York, pp.3-11 (1975); Hurley and Needham-VanDevanter, Acc.Chem.Res., 19, 230-237 (1986)). The ability of PBDs to form adducts in the minor groove allows them to interfere with DNA processing, making them suitable for use as antitumor agents. The biological activity of these molecules can be enhanced by the unification of two PBD units via a flexible alkylene linker through a C8 / C'-hydroxyl functional group (Bose, DS, et al., J. Am. Chem. Soc., 114, 4939-4941 (1992); Thurston, DE, et al., J. Org. Chem., 61, 8141-8147 (1996)). PBD dimers are thought to form sequence-selective DNA lesions such as the palindromic 5'-Pu-GATC-Py-3' interstrand crosslinking (Smellie, M., et al., Biochemistry, 42, 8232-8239 (2003); Martin, C., et al., Biochemistry, 44, 4135-4147), and this is thought to be the main mechanism governing their biological activity. An example of a PBD dimer is SG2000 (SJG-136): [ka] (Gregson, S., et al., J. Med. Chem., 44, 737-748 (2001); Alley, MC, et al., Cancer Research, 64, 6700-6706 (2004); Hartley, JA, et al., Cancer Research, 64, 6693-6699 (2004)). Due to the way these potent compounds act to crosslink DNA, conventional PBD dimers have been constructed symmetrically (i.e., both monomers of the dimer are identical). This synthetic route offers a convenient synthesis by either simultaneously constructing a PBD dimer portion with a pre-formed dimer linkage or reacting a pre-constructed PBD monomer portion with a dimer linkage group. These synthetic approaches have limited the options for preparing PBD-containing targeted conjugates. However, given the proven efficacy of PBD dimers, there is a need for asymmetric PBD dimers that can be conjugated with targeting agents for use in targeted therapy. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] Japanese Patent Application Publication No. 58-180487 [Non-patent literature]

[0007] [Non-Patent Document 1] Leimgruber, et al., J. Am. Chem. Soc., 87, 5793-5795 (1965) [Non-Patent Document 2] Leimgruber, et al., J. Am. Chem. Soc., 87, 5791-5793 (1965) [Non-Patent Document 3] Thurston,et al.,Chem.Rev.1994,433-465(1994) [Non-Patent Document 4] Hochlowski, et al., J. Antibiotics, 40, 145-148 (1987)

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[0008] 5. Outline of the Invention In this specification, compounds of formula (I): [ka] Alternatively, a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof is provided. In the formula, each of ring A and ring B is independently one of the following formulas:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0009] In this specification, compounds of formula B(i) or B(ii): [ka] Alternatively, pharmaceutically acceptable salts, tautomers, solvates, or stereoisomers thereof are also provided. Each of rings A and B is independently one of the following equations: [ka] [ka] [ka] This indicates the linking point to the linker or Ab linker. The linker is -(CH2) r -,-(CH2) p -X-(CH2) q -, or -(CH2) p -CH=CH-(CH2) q -and, X is NR 6 , NHC(=O), C(=O)NH, O, SO2, substituted or unsubstituted aryl rings, substituted or unsubstituted heteroaryl rings, substituted or unsubstituted heterocycles, or substituted or unsubstituted cyclic rings, Ring C is a cyclopropyl ring or a cyclobutyl ring. -C(R 1 )- and -N(R 2 )- Dotted line connection between [ka] Each of these is independently a single bond or a double bond, If the dotted line connection is a single connection, each R 1 Each R is independently either H or OH, and each R 2 H is H, If the dotted line bond is a double bond, then each R 1 H is independent of each R 2 It does not exist, R 3 and R 4 Each of these is independently H, NH2, and NR a R b OH, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, R a and R b These are H or C, respectively, independently. 1~4 It is alkyl, R 5 H, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, R 6 is H, or C 1~4 It is alkyl, Each of m, n, and o is independently either 1 or 2. Each of r, p, and q is an independent integer between 1 and 8. The sum of p and q is an integer between 1 and 8. An Ab linker is a compound that can link ring A or ring B to a binder.

[0010] In this specification, the conjugate of formula A(i) or A(ii): [ka] Alternatively, pharmaceutically acceptable salts, tautomers, solvates, or stereoisomers thereof are also provided. Each of rings A and B is independently one of the following equations: [ka] [ka] [ka] This indicates the linking point to the linker or Ab linker. The linker is -(CH2) r -,-(CH2) p -X-(CH2) q -, or -(CH2) p -CH=CH-(CH2) q -and, X is NR 6 , NHC(=O), C(=O)NH, O, SO2, substituted or unsubstituted aryl rings, substituted or unsubstituted heteroaryl rings, substituted or unsubstituted heterocycles, or substituted or unsubstituted cyclic rings, Ring C is a cyclopropyl ring or a cyclobutyl ring. -C(R 1 )- and -N(R 2 Each of the dotted line connections between ) and is independently a single or double bond. If the dotted line connection is a single connection, each R 1 Each R is independently either H or OH, and each R 2 H is independent of H, If the dotted line bond is a double bond, then each R 1 H is independent of each R 2 It is independently non-existent, R 3 and R 4 Each of these is independently H, NH2, and NR a R b OH, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, and R a and Rb These are H or C, respectively, independently. 1~4 It is alkyl, R 5 H, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, R 6 is H, or C 1~4 It is alkyl, Each of m, n, and o is independently either 1 or 2. Each of r, p, and q is an independent integer between 1 and 8. The sum of p and q is an integer between 1 and 8. An Ab linker is a compound that links Ab to ring A or ring B. Ab is a binder selected from humanized antibodies, chimeric antibodies, or human antibodies, or their antigen-binding fragments. The subscript x is between 1 and 15. 6. Brief explanation of the drawing [Brief explanation of the drawing]

[0011] [Figure 1] This is a line graph showing the killing of A375 cells by the compounds disclosed herein. [Figure 2] This is a line graph showing the killing of Calu-6 cells by the compounds disclosed herein. [Figure 3] This is a line graph showing the killing of A375 cells by the compounds disclosed herein. [Figure 4] This is a line graph showing the killing of Calu-6 cells by the compounds disclosed herein. [Figure 5] This is a line graph showing the killing of A375 cells by the compounds disclosed herein. [Figure 6] This is a line graph showing the killing of Calu-6 cells by the compounds disclosed herein. [Figure 7] This is a line graph showing the killing of A375 cells by the compounds disclosed herein. [Figure 8]This is a line graph showing the killing of Calu-6 cells by the compounds disclosed herein. [Figure 9] This is a line graph showing the killing of A375 cells by the compounds disclosed herein. [Figure 10] This is a line graph showing the killing of Calu-6 cells by the compounds disclosed herein. [Figure 11] This is a line graph showing the killing of A375 cells by the compounds disclosed herein. [Figure 12] This is a line graph showing the killing of Calu-6 cells by the compounds disclosed herein. [Figure 13] This is a line graph showing the killing of A375 cells by the compounds disclosed herein. [Figure 14] This is a line graph showing the killing of Calu-6 cells by the compounds disclosed herein. [Modes for carrying out the invention]

[0012] 7. Modes for Carrying Out the Invention This specification provides pyrrolobenzodiazepine (PBD) compounds comprising two PBDs, which may be identical or different, linked by a linker. These compounds can be used as the drug or payload portion of an antibody-drug conjugate (ADC). Diseases or disorders, such as cancer, can be treated using ADCs, for example, by providing compositions comprising ADCs.

[0013] 7.1.Definition In this disclosure, the following terms have the meanings set forth below unless otherwise indicated. Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by those skilled in the art in which this disclosure pertains. If there are multiple definitions of a term used herein, these definitions shall prevail unless otherwise noted.

[0014] Where a trademark is used herein, unless the context otherwise specifies, a reference to the trademark name also refers to the product formulations, generic drugs, and active pharmaceutical ingredients of the trademarked product.

[0015] In this specification, the term “antibody” is used in its broadest sense and specifically encompasses intact monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments exhibiting desired biological activity. Intact antibodies primarily have two regions: a variable region and a constant region. The variable region binds to and interacts with the target antigen. The variable region includes a complementary determination region (CDR) that recognizes and binds to a specific binding site on a particular antigen. The constant region is recognized by the immune system and can interact with it (see, e.g., Janeway et al., 2001, Immuno. Biology, 5th Ed., Garland Publishing, New York). Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass. Antibodies can be derived from any suitable species. In some embodiments, the antibodies are derived from humans or mice. The antibodies may be, for example, human, humanized, or chimeric antibodies.

[0016] As used herein, the term “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous antibody population (i.e., the individual antibodies constituting the population are identical except for minor naturally occurring variations that may be present). Monoclonal antibodies are highly specific and target a single antigenic site. The modifier “monoclonal” should not be interpreted as requiring antibody production by any particular method.

[0017] An "intact antibody" is an antibody that, in addition to an antigen-binding variable region, contains a light chain constant domain (CL) and a heavy chain constant domain (CH1, CH2, CH3, and CH4) (depending on the antibody class). The constant domain may be a native sequence constant domain (e.g., a human native sequence constant domain) or an amino acid sequence variant thereof.

[0018] An "antibody fragment" includes a portion of an intact antibody containing an antigen-binding region or a variable region. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments, diabodies, triabodies, tetrabodies, linear antibodies, single-chain antibody molecules, scFv, scFv-Fc, antibody fragments (multiple), multispecific antibody fragments formed from fragments (multiple) produced by a Fab expression library, or any of the above epitope-binding fragments that bind immunospecifically to a target antigen (e.g., cancer cell antigen, viral antigen, or microbial antigen).

[0019] An "antigen" is the substance to which an antibody specifically binds.

[0020] The terms "specific binding" and "specifically binding" mean that an antibody or antibody derivative binds to its corresponding target antigen in a highly selective manner and does not bind to a large number of other antigens. Typically, an antibody or antibody derivative binds to at least about 1 × 10⁻¹⁶ antigens. -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, or 10 -12 It binds with affinity M, and binds to the given antigen with an affinity at least twice as great as the affinity it has for binding to the given antigen or nonspecific antigens other than the given antigen or closely related antigens (e.g., BSA, casein).

[0021] The terms "inhibit" or "inhibit" mean to reduce a measurable amount or to completely prevent it.

[0022] The term "therapeutic dose" refers to the amount of a drug effective in treating a disease or disorder in a mammal. In the case of cancer, a therapeutic dose of a drug can reduce the number of cancer cells, shrink tumor size, inhibit (i.e., slow or stop to some extent) cancer cells from invading peripheral organs, inhibit (i.e., slow or stop to some extent) tumor metastasis, inhibit tumor growth to some extent, and / or alleviate to some extent one or more symptoms associated with cancer. A drug has cell division inhibitory and / or cytotoxic properties to the extent that it can inhibit growth and / or kill existing cancer cells. In the case of cancer treatment, efficacy can be measured, for example, by evaluating the time to progression (TTP) and / or determining the response rate (RR).

[0023] The terms “substantial” or “substantially” refer to the majority of a mixture or sample population, i.e., >50%, preferably 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or more than 99% of the population.

[0024] The terms "intracellularly cleaved" and "intracellular cleavage" refer to intracellular metabolic processes or reactions to ligand-drug conjugates (e.g., antibody-drug conjugates (ADCs)) where the covalent bond between the drug portion (D) and the ligand unit (e.g., antibody (BA or Ab)), such as the linker, is cleaved, resulting in the release of the drug or another metabolite of the conjugate within the cell. Therefore, the cleaved portion of a drug-linker-ligand conjugate is an intracellular metabolite.

[0025] The terms “cancer” and “malignant” refer to or describe a physiological condition or disorder in mammals typically characterized by uncontrolled cell proliferation. A “tumor” contains one or more cancerous cells. Examples of cancer include, but are not limited to, carcinomas, lymphomas, blastomas, sarcomas, and leukemias or lymphoid malignancies. More specific examples of such cancers include squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer (including small cell lung cancer, non-small cell lung cancer ("NSCLC"), lung adenocarcinoma, and lung squamous cell carcinoma), peritoneal cancer, hepatocellular carcinoma, gastric cancer or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatocellular carcinoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, and head and neck cancer.

[0026] In this specification, “autoimmune disease” means a disease or disorder that originates from and is directed towards the tissues or proteins of an individual.

[0027] Examples of “patient” or “subject” include, but are not limited to, mammals (e.g., humans, rats, mice, guinea pigs, monkeys, pigs, goats, cattle, horses, dogs, or cats), and birds or poultry. In one embodiment, the patient is human.

[0028] The terms “to treat” or “treatment” refer to therapeutic treatment and preventive measures to prevent recurrence, unless otherwise indicated by the context, and their purpose is to suppress or slow (mitigate) undesirable physiological changes or impairments (e.g., the development or spread of cancer). For the purposes of this disclosure, beneficial or desirable clinical outcomes include, but are not limited to, symptom relief, reduction of disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or mitigation of the disease state, and remission (partial or overall), whether detectable or undetectable. “Treatment” may also mean extending survival compared to the survival expected without treatment. Those requiring treatment include those who already have a condition or impairment, as well as those who are prone to developing a condition or impairment.

[0029] In the context of cancer, the term “to treat” includes any or all of the following: inhibiting tumor cells, cancer cells, or tumor growth; inhibiting the replication of tumor cells or cancer cells; reducing the overall tumor burden or decreasing the number of cancer cells; and improving one or more symptoms associated with the disease.

[0030] In the context of autoimmune diseases, the term “treat” includes any or all of the following: inhibiting the replication of cells associated with the autoimmune disease (including, but not limited to, cells that produce autoimmune disease antibodies); reducing the autoimmune antibody load; and improving one or more symptoms of the autoimmune disease.

[0031] As used herein and in the appended claims, the indefinite articles "a" and "an," and the definite article "the," include singular as well as plural referents, unless otherwise explicitly indicated by the context.

[0032] As used herein, and unless otherwise specified, the terms “about” and “approximately” mean, when used in relation to the amount or weight percentage of a component of a composition, an amount or weight percentage that would be recognized by those skilled in the art as producing an equivalent pharmacological effect to that obtained from a specified amount or weight percentage. In certain embodiments, when used in this context, the terms “about” and “approximately” mean an amount or weight percentage that is within 30%, 20%, 15%, 10%, or 5% of a specified amount or weight percentage.

[0033] As used herein, unless otherwise specified, the terms “about” and “approximately” mean, when used in relation to a numerical value or range of values ​​provided to characterize a particular solid form, e.g., a specific temperature or temperature range, e.g., one describing melting, dehydration, desolvation, or glass transition temperature; a mass change, e.g., a mass change as a function of temperature or humidity; a solvent or water content (e.g., in terms of mass or percentage); or a peak position, e.g., in analysis by IR, Raman spectroscopy, or XRPD, that the value or range of values ​​may deviate to an extent that is reasonable to those skilled in the art, but still describe a solid form. Techniques for characterizing crystalline morphology and amorphous solids include, but are not limited to, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), single-crystal X-ray diffraction, vibrational spectroscopy, such as infrared (IR) and Raman spectroscopy, solid-state and solution nuclear magnetic resonance (NMR) spectroscopy, optical microscopy, hot-stage optical microscopy, scanning electron microscopy (SEM), electron crystallography and quantitative analysis, particle size analysis (PSA), surface area analysis, solubility studies, and dissolution studies. In certain embodiments, the terms “about” and “approximately” used in this context indicate that a numerical value or range may vary within 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.5%, 1%, 0.5%, or 0.25% of the given numerical value or range. For example, in some embodiments, the value of the XRPD peak position may vary by up to ±0.2°²θ while describing a particular XRPD peak.

[0034] The "alkyl" group is a saturated, partially saturated, or unsaturated linear or branched acyclic hydrocarbon having 1 to 10 carbon atoms, typically 1 to 8 carbon atoms, and in some embodiments 1 to 6, 1 to 4, or 2 to 6 carbon atoms. Representative alkyl groups include -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl. Saturated branched alkyl groups include -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, and 2,3-dimethylbutyl. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, allyl, -CH=CH(CH3), -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), C(CH2CH3)=CH2, -C≡CH, -C≡C(CH3), -C≡C(CH2CH3), -CH2C≡CH, -CH2C≡C(CH3), and CH2C≡C(CH2CH3). Alkyl groups may be substituted or unsubstituted. In certain embodiments, where an alkyl group described herein is expressed as “substituted,” the alkyl group may be substituted with halogens (chloro, iodine, bromo, or fluoro), hydroxyl, alkoxy, alkoxyalkyl, amino, alkylamino, carboxy, nitro, cyano, thiol, thioether, imine, imide, amidine, guanidine, enamine, aminocarbonyl, acylamino, phosphonato, phosphine, thiocarbonyl, sulfonyl, sulfone, sulfonamide, ketone, aldehyde, ester, urea, urethane, oxime, hydroxylamine, alkoxyamine, aralkoxyamine, N-oxide, hydrazine, hydrazide, hydrazone, azide, isocyanate, isothiocyanate, cyanate, thiocyanate, B(OH)2, or O(alkyl)aminocarbonyl, in addition to any substituent(s) shown in the compounds and embodiments disclosed herein.

[0035] The "alkenyl" group is a linear or branched acyclic hydrocarbon having 2 to 10 carbon atoms, typically 2 to 8 carbon atoms, and containing at least one carbon-carbon double bond. Representative linear and branched (C2-C8) alkenyls include -vinyl, -allyl, -1-butenyl, -2-butenyl, -isobutyrenyl, -1-pentenyl, -2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, -2,3-dimethyl-2-butenyl, -1-hexenyl, 2-hexenyl, -3-hexenyl, -1-heptenyl, -2-heptenyl, -3-heptenyl, -1-octenyl, -2-octenyl, and 3-octenyl. The double bond of the alkenyl group may or may not conjugate with another unsaturated group. The alkenyl group may be unsubstituted or substituted.

[0036] The "cycloalkyl" group is a saturated or partially saturated cyclic alkyl group of 3 to 10 carbon atoms having a single cyclic ring or multiple fused or crosslinked rings, and may optionally be substituted with 1 to 3 alkyl groups. In some embodiments, the cycloalkyl group has 3 to 8 ring members, while in other embodiments, the number of ring carbon atoms is in the range of 3 to 5, 3 to 6, or 3 to 7. Examples of such cycloalkyl groups include monocyclic structures (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, etc.) or polycyclic or crosslinked ring structures (e.g., adamantyl, etc.). Examples of unsaturated cycloalkyl groups include, in particular, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl. The cycloalkyl group may be substituted or unsubstituted. Examples of such substituted cycloalkyl groups include cyclohexanone.

[0037] An "aryl" group is an aromatic carbocyclic group with 6 to 14 carbon atoms, having a single ring (e.g., phenyl) or multiple fused rings (e.g., naphthyl or anthryl). In some embodiments, the aryl group contains 6 to 14 carbon atoms in the ring portion of the group, and in others, 6 to 12, and possibly 6 to 10 carbon atoms. Specific examples of aryls include phenyl, biphenyl, and naphthyl. The aryl group may be substituted or unsubstituted. The term "aryl group" also includes groups containing fused rings, such as aromatic-aliphatic fused ring systems (e.g., indanyl, tetrahydronaphthyl, etc.).

[0038] The "arylene" group is a divalent aryl group as defined herein.

[0039] A "heteroaryl" group is an aryl ring system having 1 to 4 heteroatoms as ring atoms in a heteroaromatic ring system, and the remaining atoms are carbon atoms. In some embodiments, the heteroaryl group contains 5 to 6 ring atoms in the ring portion of the group, and in other cases contains 6 to 9 or 6 to 10 atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen. In certain embodiments, the heteroaryl ring system is monocyclic or bicyclic. Non-limiting examples include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyrrolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiophenyl, benzothiophenyl, furanyl, benzofuranyl (e.g., isobenzofuran-1,3-diamine), indolyl, azaindolyl (e.g., pyrrolopyridyl or 1H-pyrrolo[2,3-b]pyridyl), indazolyl, benzimidazolyl (e.g., 1H-benzo[d]imidazolyl), imidazopyridyl (e.g., azabenzimidazolyl, 3H-imidazo[4,5-b]pyridyl, or 1H-imidazo[4,5-b]pyridyl), pyrazolopyridyl, triazolopyridyl, benzotriazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, isoxazolopyridyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups, etc.

[0040] A "heteroarylene" group is a divalent heteroaryl group as defined herein.

[0041] A "heterocyclyl" is an aromatic (also called heteroaryl) or non-aromatic cycloalkyl group in which 1 to 4 of the ring carbon atoms are independently replaced by heteroatoms from the group consisting of O, S, and N. In some embodiments, the heterocyclyl group has 3 to 10 ring members, while other such groups have 3 to 5, 3 to 6, or 3 to 8 ring members. Furthermore, the heterocyclyl may be bonded to other groups by any ring atom (i.e., any carbon or heteroatom of the heterocyclic ring). The heteroaryl group may be substituted or unsubstituted. Heterocyclyl groups include unsaturated, partially saturated, and saturated ring systems (e.g., imidazolyl, imidazolinyl, and imidazolidinyl groups). The term "heterocyclyl" includes fused ring species (including ring species containing fused aromatic and non-aromatic groups), such as benzotriazolyl, 2,3-dihydrobenzo[1,4]dioxynyl, and benzo[1,3]dioxolyl. The term also includes bridging polycyclic ring systems containing heteroatoms (for example, quinuclidyl, but not limited to these).Examples of heterocyclyl groups, though not limited to them, include azilidinyl, azetidinyl, pyrrolidyl, imidazolidinyl, pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl, tetrahydrofuranil, dioxolyl, furanil, thiophenyl, pyrrolyl, pyrrolinil, imidazolyl, pyrazolyl, pyrazolinil, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, thiazolinyl, isothiazolyl, thiadiazolyl, oxadiazolyl, piperidyl, piperazinyl, morpholinil, and thiomorpholinyl. Tetrahydropyranil (e.g., tetrahydro-2H-pyranil), tetrahydrothiopyranil, oxatian, dioxyl, dithianil, pyranil, pyridyl, pyrimidinil, pyridazinil, pyrazinil, triazinil, dihydropyridyl, dihydrodithinyl, dihydrodithionyl, homopiperazinil, quinuclidyl, indolyl, indolinyl, isoindolyl, azaindolyl (pyrrolopyridyl), indazolyl, indolidinyl, benzotriazolyl, benzimidazolyl, benzofuranil, benzothiophenyl, benzothiazolyl Benzoxadiazolyl, benzoxazinyl, benzodithinyl, benzoxathinyl, benzothiadinyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[1,3]dioxolyl, pyrazolopyridyl, imidazopyridyl (azabenzimidazolyl, e.g., 1H-imidazo[4,5-b]pyridyl or 1H-imidazo[4,5-b]pyridin-2(3H)-onyl), triazolopyridyl, isoxazolopyridyl, prinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, ki Examples include noridinyl, quinoxalinyl, quinazolinyl, cinolinyl, phthalazinyl, naphthilidinyl, pteridinyl, thianaphthalenyl, dihydrobenzothiadinyl, dihydrobenzofuranyl, dihydroindolyl, dihydrobenzodioxynyl, tetrahydroindolyl, tetrahydroindazolyl, tetrahydrobenzimidazolyl, tetrahydrobenzotriazolyl, tetrahydropyrrolopyridyl, tetrahydropyrazolopyridyl, tetrahydroimidazopyridyl, tetrahydrotriazolopyridyl, and tetrahydroquinolinyl groups.Representative substituted heterocyclic groups include, for example, but not limited to, pyridyl or morpholinyl groups that are mono-substituted or polysubstituted with various substituents such as those listed below, or are 2-, 3-, 4-, 5-, or 6-substituted.

[0042] The "cycloalkylalkyl" group is a radical of the formula: -alkyl-cycloalkyl (where alkyl and cycloalkyl are defined above). The substituted cycloalkylalkyl group may be substituted in the alkyl part, cycloalkyl part, or both the alkyl part and cycloalkyl part of this group. Representative cycloalkylalkyl groups include, but are not limited to, cyclopentylmethyl, cyclopentylethyl, cyclohexylmethyl, cyclohexylethyl, and cyclohexylpropyl. Representative substituted cycloalkylalkyl groups may be mono-substituted or polysubstituted.

[0043] The "aralkyl" group is a radical of the formula: -alkyl-aryl (where alkyl and aryl are defined above). The substituted aralkyl group may be substituted in the alkyl part, aryl part, or both the alkyl part and aryl part of the group. Representative aralkyl groups include, but are not limited to, benzyl and phenethyl groups, and fused (cycloalkylaryl) alkyl groups (e.g., 4-ethyl-indanyl).

[0044] The "heterocyclylalkyl" group is a radical of the formula :-alkyl-heterocyclyl (wherein alkyl and aryl are defined above). Substituted heterocyclylalkyl groups can be substituted with the alkyl moiety, the heterocyclyl moiety, or both the alkyl and heterocyclyl moieties of the group. Representative heterocyclylalkyl groups include, but are not limited to, 4-ethyl-morpholinyl, 4-propylmorpholinyl, furan-2-ylmethyl, furan-3-ylmethyl, pyrzin-3-ylmethyl, (tetrahydro-2H-pyran-4-yl)methyl, (tetrahydro-2H-pyran-4-yl)ethyl, tetrahydrofuran-2-ylmethyl, tetrahydrofuran-2-ylethyl, and indole-2-ylpropyl.

[0045] "Halogen" refers to chloro, iodine, bromo, or fluoro compounds.

[0046] A "hydroxyalkyl" group is one in which the aforementioned alkyl group is replaced with one or more hydroxyl groups.

[0047] The "alkoxy" group is O(alkyl) (wherein alkyl is defined above).

[0048] The "alkoxyalkyl" group is -(alkyl)-O-(alkyl) (where alkyl is defined above).

[0049] As used herein, “alkynyl” refers to a monovalent hydrocarbon radical moiety comprising at least two carbon atoms and one or more carbon-carbon triple bonds. Alkynnyls can be optionally substituted and may be linear, branched, or cyclic. Alkynnyls are radicals having 2 to 20 carbon atoms, i.e., C 2-20 Alkynyl radicals are radicals having 2 to 12 carbon atoms, i.e., C 2-12 Alkynyl radicals are radicals having 2 to 8 carbon atoms, i.e., C 2-8 Alkynyl radicals are radicals having 2 to 6 carbon atoms, i.e., C 2-6Alkynyl radicals, and radicals having 2 to 4 carbon atoms, i.e., C 2-4 This includes, but is not limited to, alkynyl radicals. Examples of alkynyl moieties include, but are not limited to, ethynyl, propynyl, and butynyl.

[0050] As used herein, “haloalkyl” refers to the alkyl group as defined above, where the alkyl group comprises at least one substituent selected from halogens, such as fluorine (F), chlorine (Cl), bromine (Br), or iodine (I). Examples of haloalkyl groups, but not limited to, include -CF3, -CH2CF3, -CCl2F, and -CCl3.

[0051] As used herein, “haloalkoxy” means the alkoxy as defined above, where the alkoxy comprises at least one substituent selected from a halogen, e.g., F, Cl, Br, or I.

[0052] As used herein, “arylalkyl” refers to the monovalent radical portion of an alkyl compound, and the alkyl compound is substituted with an aromatic substituent. That is, the aromatic compound contains a single bond to the alkyl group, and the radical is localized to the alkyl group. The arylalkyl group is bonded to the shown chemical structure via the alkyl group. The arylalkyl group can be represented by the structure, for example, B-CH2-, B-CH2-CH2-, B-CH2-CH2-CH2-, B-CH2-CH2-CH2-CH2-, B-CH(CH3)-CH2-CH2-, B-CH2-CH(CH3)-CH2-, where B is an aromatic moiety, for example, phenyl. The arylalkyl group is optionally substituted; that is, the aryl group and / or alkyl group can be substituted as disclosed herein. Examples of arylalkyl groups, but not limited to, benzyl.

[0053] As used herein, "alkylaryl" refers to a monovalent moiety that is a radical of an aryl compound, where the aryl compound is substituted with an alkyl substituent. That is, the aryl compound contains a single bond to an alkyl group, and the radical is localized on the aryl group. The alkylaryl group is bonded to the indicated chemical structure through the aryl group. Alkylaryl can be represented by structures such as -B-CH3, -B-CH2-CH3, -B-CH2-CH2-CH3, -B-CH2-CH2-CH2-CH3, -B-CH(CH3)-CH2-CH3, -B-CH2-CH(CH3)-CH3, where B is an aromatic moiety, such as phenyl. Alkylaryl is optionally substituted. That is, the aryl group and / or the alkyl group can be substituted as disclosed herein. Examples of alkylaryl include, but are not limited to, toluyl.

[0054] As used herein, "aryloxy" refers to a monovalent moiety that is a radical of an aromatic compound, where the ring atoms are carbon atoms and the ring is substituted with an oxygen radical, i.e., the aromatic compound contains a single bond to an oxygen atom and the radical is localized on the oxygen atom (e.g., in the case of phenoxy, C6H5-O-). The aryloxy substituent is bonded to the compound being substituted through this oxygen atom. Aryloxy is optionally substituted. Aryloxy includes, but is not limited to, radicals having 6 to 20 ring carbon atoms, i.e., C 6~20 aryloxy radicals, radicals having 6 to 15 ring carbon atoms, i.e., C 6~15 aryloxy radicals, and radicals having 6 to 10 ring carbon atoms, i.e., C 6~10 aryloxy radicals. Examples of aryloxy moieties include, but are not limited to, phenoxy, naphthoxy, and anthroxy.

[0055] The "amino" group is a radical of the formula: NH2.

[0056] The "hydroxylamine" group has the formula: N(R #)a radical of OH or NHOH, wherein R # is a substituted or unsubstituted alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, or heterocyclylalkyl group as defined herein).

[0057] The “alkoxyamine” group is a radical of the formula: -N(R # )O-alkyl or -NHO-alkyl, wherein R # is as defined above).

[0058] The “aralkoxyamine” group is a radical of the formula: N(R # )O-aryl or NHO aryl, wherein R # is as defined above).

[0059] The “alkylamine” group is a radical of the formula: NH alkyl or N(alkyl)2, wherein each alkyl is independently as defined above).

[0060] The “aminocarbonyl” group is a radical of the formula: -C(=O)N(R # )2, -C(=O)NH(R # ), or C(=O)NH2, wherein each R # is as defined above).

[0061] The “acylamino” group is a radical of the formula: NHC(=O)(R # ) or N(alkyl)C(=O)(R # ), wherein each alkyl and R # are independently as defined above).

[0062] The “O(alkyl)aminocarbonyl” group is a radical of the formula: -O(alkyl)C(=O)N(R # )2, -O(alkyl)C(=O)NH(R # ), or -O(alkyl)C(=O)NH2, wherein each R #is, independently, as defined above).

[0063] The "N-oxide" group is a radical of the formula: -N + -O - .

[0064] The "carboxy" group is a radical of the formula: -C(O)OH.

[0065] The "ketone" group is a radical of the formula: C(=O)(R # )(wherein R # is, independently, as defined above).

[0066] The "aldehyde" group is a radical of the formula: -CH(=O).

[0067] The "ester" group is a radical of the formula: C(=O)O(R # ) or OC(=O)(R # )(wherein R # is, independently, as defined above).

[0068] The "urea" group is a radical of the formula: -N(alkyl)C(=O)N(R # )2, -N(alkyl)C(=O)NH(R # ), -N(alkyl)C(=O)NH2, -NHC(=O)N(R # )2, -NHC(=O)NH(R # ), or NHC(=O)NH2 # (wherein each alkyl and R # is, independently, as defined above).

[0069] The "imine" group is a radical of the formula: -N=C(R # )2 or -C(R # )=N(R # )(wherein each R # is, independently, as defined above).

[0070] The "imide" group is a radical of the formula: -C(=O)N(R#)C(=O)(R# ) or N((C=O)(R # ))2 radicals (where each R # (These are independent, as defined above.)

[0071] The "urethane" group is represented by the formula: -OC(=O)N(R # )2, -OC(=O)NH(R # ), -N(R # )C(=O)O(R # ), or -NHC(=O)O(R # ) is a radical (where each R # (These are independent, as defined above.)

[0072] The "amidine" group is represented by the formula: -C(=N(R # ))N(R # )2, -C(=N(R # ))NH(R # ), -C(=N(R # ))NH2, -C(=NH)N(R # )2, -C(=NH)NH(R # ), -C(=NH)NH2, -N=C(R # )N(R # )2, -N=C(R # )NH(R # ), -N=C(R # )NH2, -N(R # )C(R # )=N(R # ), -NHC(R # )=N(R # ), -N(R # )C(R # )=NH, or -NHC(R # ) = NH radical (where R # (These are independent, as defined above.)

[0073] The "guanidine" group is represented by the formula: -N(R # )C(=N(R # ))N(R # )2, -NHC(=N(R # ))N(R # )2, -N(R# )C(=NH)N(R # )2, -N(R # )C(=N(R # ))NH(R # ), -N(R # )C(=N(R # ))NH2, -NHC(=NH)N(R # )2, -NHC(=N(R # ))NH(R # ), -NHC(=N(R # ))NH2, -NHC(=NH)NH(R # ), -NHC(=NH)NH2, -N=C(N(R # )2)2, -N=C(NH(R # ))2, or the radical of -N=C(NH2)2 (where R is the radical in the formula). # (These are independent, as defined above.)

[0074] The "enamine" group is represented by the formula: -N(R # )C(R # )=C(R # )2, -NHC(R # )=C(R # )2, -C(N(R # )2)=C(R # )2, -C(NH(R # ))=C(R # )2, -C(NH2)=C(R # )2, -C(R # )=C(R # )(N(R # )2), C(R # )=C(R # )(NH(R # )) or -C(R # )=C(R # The radical of )(NH2) (where R is in the formula) # (These are independent, as defined above.)

[0075] The "oxime" group is represented by the formula: -C(=NO(R # ))(R # ), -C(=NOH)(R # ), -CH(=NO(R #)), or a radical of -CH(=NOH) (where each R # (These are independent, as defined above.)

[0076] The "hydrazide" group is represented by the formula: -C(=O)N(R # )N(R # )2, -C(=O)NHN(R # )2, -C(=O)N(R # )NH(R # ) 、 -C(=O)N(R # )NH2, -C(=O)NHNH(R # )2, or the radical of -C(=O)NHNH2 (where R is the radical in the formula) # (These are independent, as defined above.)

[0077] The "hydrazine" group is represented by the formula: -N(R # )N(R # )2, -NHN(R # )2, -N(R # )NH(R # ) 、 -N(R # )NH2, -NHNH(R # )2, or the radical of -NHNH2 (wherein R, R # (These are independent, as defined above.)

[0078] The "hydrazone" group is represented by the formula: -C(=NN(R # )2)(R # )2, -C(=NNH(R # ))(R # )2, -C(=N-NH2)(R # )2, -N(R # )(N=C(R # )2), or -NH(N=C(R # )2) is a radical (where R is in the formula) # (These are independent, as defined above.)

[0079] The "azide" group is a radical of formula -N3.

[0080] The "isocyanate" group is a radical with the formula N=C=O.

[0081] The "isothiocyanate" group is a radical with the formula N=C=S.

[0082] The "cyanate" group is the radical of formula OCN.

[0083] The "thiocyanate" group is a radical of formula SCN.

[0084] The "thioether" group is represented by the formula: -S(R # ) is a radical (where R is in the formula) # (This is as defined above).

[0085] The "thiocarbonyl" group is represented by the formula: -C(=S)(R # ) is a radical (where R is in the formula) # (These are independent, as defined above.)

[0086] The "sulfinyl" group is represented by the formula: -S(=O)(R # ) is a radical (where R is in the formula) # (These are independent, as defined above.)

[0087] The "sulfone" group is represented by the formula: -S(=O)2(R # ) is a radical (where R is in the formula) # (These are independent, as defined above.)

[0088] The "sulfonylamino" group is represented by the formula -NHSO2(R # ) or -N(alkyl)SO2(R # ) is a radical, where each alkyl and R # This is defined above.

[0089] The "sulfonamide" group is represented by the formula -S(=O)2N(R # )2, or -S(=O)2NH(R # ), or -S(=O)2NH2 radical, where each R #These are defined independently as described above.

[0090] The "phosphonate" group is represented by the formula -P(=O)(O(R # ))2, -P(=O)(OH)2, -OP(=O)(O(R # ))(R # ), or -OP(=O)(OH)(R # ) is a radical, in the formula, each R # These are defined independently as described above.

[0091] The "phosphine" group is represented by the formula: -P(R # )2 radicals (where each R # (These are independent, as defined above.)

[0092] Where a group described herein (with the exception of alkyl groups) is expressed as "substituted," that group may be substituted with any suitable substituent(s) (one or more). Exemplary examples of substituents include those found in the compounds and embodiments disclosed herein, as well as halogens (chloro, iodo, bromo, or fluoro), alkyl, hydroxyl, alkoxy, alkoxyalkyl, amino, alkylamino, carboxy, nitro, cyano, thiol, thioether, imine, imide, amidine, guanidine, enamine, aminocarbonyl, acylamino, phosphonate, phosphine, thiocarbonyl, sulfinyl, sulfone, sulfonamide, ketone, aldehyde, ester, urea, urethane, oxime, hydroxylamine, alkoxyamine, aralkoxyamine, N-oxide, hydrazine, hydrazide, hydrazone, azide, isocyanate, isothiocyanate, cyanate, thiocyanate, oxygen (=O), B(OH)2, O(alkyl)aminocarbonyl, monocyclic, condensed or These include cycloalkyl compounds that may be non-condensed polycyclic (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or heterocyclil compounds that may be monocyclic, condensed, or non-condensed polycyclic (e.g., pyrrolidyl, piperidyl, piperazinyl, morpholinyl, or thiadinyl), monocyclic, condensed, or non-condensed polycyclic aryl or heteroaryl compounds (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanil, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridinyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridadinyl, pyrimidinyl, benzimidazolyl, benzothiophenyl, or benzofuranil), aryloxys, aralkyloxys, heterocyclyloxys, and heterocyclylalkoxys.

[0093] As used herein, “pharmaceutically acceptable salts” means salts prepared from pharmaceutically acceptable, non-toxic acids or bases, including inorganic acids or bases and organic acids or bases.

[0094] As used herein, unless otherwise indicated, the term “solvate” means a compound or a salt thereof that further contains a stoichiometric or nonstoichiometric amount of solvent bonded by non-covalent intermolecular forces. In one embodiment, the solvate is a hydrate.

[0095] As used herein, unless otherwise indicated, the term “hydrate” means a compound or salt thereof that further contains stoichiometric or nonstoichiometric amounts of water bound by non-covalent intermolecular forces.

[0096] As used herein, unless otherwise indicated, the term “prodrug” means a compound derivative that can be hydrolyzed, oxidized, or otherwise reacted under biological conditions (in vitro or in vivo) to provide an active compound. Examples of prodrugs, but not limited to, are derivatives and metabolites of compounds containing biohydrolyzable sites, such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureids, and biohydrolyzable phosphate analogs. In certain embodiments, a prodrug of a compound having a carboxyl functional group is a lower alkyl ester of a carboxylic acid. Carboxylic acid esters can be formed by esterifying any of the carboxylic acid moieties present on the molecule. Typically, prodrugs are formed by well-known methods (e.g., Burger's Medicinal Chemistry and Drug Discovery 6). th It can be prepared using the methods described in (Donald J. Abraham ed., 2001, Wiley) and Design and Application of Prodrugs (H. Bundgaard ed., 1985, Harwood Academic Publishers GmbH).

[0097] As used herein, and unless otherwise indicated, the terms “stereoisomer” or “stereoisomerically pure” mean a single stereoisomer of a compound that is substantially free of other stereoisomers. For example, a stereoisomerically pure compound having one chiral center is substantially free of the opposite enantiomer of that compound. A stereoisomerically pure compound having two chiral centers is substantially free of other diastereomers of that compound. Typical stereoisomerically pure compounds contain about 80% by weight of one stereoisomer and less than about 20% by weight of other stereoisomers of that compound, about 90% by weight of one stereoisomer and less than about 10% by weight of other stereoisomers of that compound, about 95% by weight of one stereoisomer and less than about 5% by weight of other stereoisomers of that compound, or about 97% by weight of one stereoisomer and less than about 3% by weight of other stereoisomers of that compound. Compounds may have chiral centers and may exist as racemates, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms, including mixtures thereof, are included in the embodiments disclosed herein. The use of stereoisomerically pure forms of such compounds, as well as mixtures of those forms, are encompassed by the embodiments disclosed herein. For example, mixtures containing equimolar or unequal amounts of enantiomers of a particular compound may be used in the methods and compositions disclosed herein. These isomers may be synthesized asymmetrically or resolved using standard techniques such as chiral columns or chiral resolving agents.For example, see Jacques, J., et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, SH, et al., Tetrahedron 33:2725 (1977); Eliel, EL, Stereochemistry of Carbon Compounds (McGraw Hill, NY, 1962); and Wilen, SH, Tables of Resolving Agents and Optical Resolutions p.268 (EL Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972).

[0098] It should also be noted that the compound may include E and Z isomers or mixtures thereof, as well as cis and trans isomers or mixtures thereof. In certain embodiments, the compound is isolated as either a cis or trans isomer. In other embodiments, the compound is a mixture of cis and trans isomers.

[0099] A "tautomer" refers to an isomer of a compound that is in equilibrium with itself. The concentrations of these isomers vary depending on the environment in which the compound is found; for example, they may differ depending on whether the compound is in a solid state or in an organic solution or aqueous solution. For example, in aqueous solution, pyrazole may exhibit the following isomers, which are referred to as tautomers of each other: [ka]

[0100] As will be readily apparent to those skilled in the art, a wide variety of functional groups and other structures may exhibit tautomerism, and all tautomers of a compound are within the scope of this disclosure.

[0101] It should also be noted that compounds may contain unnatural proportions of atomic isotopes in one or more atoms. For example, a compound may contain tritium ( 3 H), Iodine-125( 125 I), Sulfur-35( 35 S), or carbon-14 ( 14 It may be radioactively labeled with radioactive isotopes such as C, or deuterium ( 2 H), carbon-13 ( 13 C), or nitrogen-15( 15 They may be isotope-enriched, such as N). As used herein, “isotope-substituted compound” refers to an isotope-enriched compound. The term “isotope-enriched” refers to an atom having an isotope composition other than the atom’s natural isotope composition. “Isotope-enriched” may also refer to a compound containing at least one atom having an isotope composition other than the atom’s natural isotope composition. The term “isotope composition” refers to the amount of each isotope present in a given atom. Radiolabeled and isotope-enriched compounds are useful as therapeutic agents, e.g., cancer and inflammation treatments; research reagents, e.g., binding assay reagents; and diagnostic agents, e.g., in vivo contrast agents. All isotope variations of the compounds described herein, whether radioactive or not, are intended to be included within the scope of the embodiments provided herein. In some embodiments, isotopologs of the compounds are provided, for example, isotopologs are deuterium, carbon-13, or nitrogen-15 enriched compounds.

[0102] Please note that if there is a discrepancy between the illustrated structure and its name, the illustrated structure will be given more weight.

[0103] In this specification, the term “residue” refers to a chemical portion of a compound that remains after a chemical reaction. For example, the terms “amino acid residue” or “N-alkyl amino acid residue” refer to the product of an amide coupling or peptide coupling of an amino acid or N-alkyl amino acid with a suitable coupling partner, for example, a water molecule being expelled after the amide or peptide coupling of an amino acid or N-alkyl amino acid, resulting in the incorporation of an amino acid residue or N-alkyl amino acid residue into the product.

[0104] As used herein, “sugar,” “sugar group,” or “sugar residue” refers to a carbohydrate portion that may include 3-carbon (triose) units, 4-carbon (tetrose) units, 5-carbon (pentose) units, 6-carbon (hexose) units, 7-carbon (heptose) units, or combinations thereof, and may be a monosaccharide, disaccharide, trisaccharide, tetrasaccharide, pentasaccharide, oligosaccharide, or any other polysaccharide. In some cases, “sugar,” “sugar group,” or “sugar residue” may include furanoses (e.g., ribofuranose, fructofuranose) or pyranoses (e.g., glucopyranose, galactopyranose), or combinations thereof. In some cases, “sugar,” “sugar group,” or “sugar residue” may include aldoses or ketoses, or combinations thereof. Non-limiting examples of monosaccharides include ribose, deoxyribose, xylose, arabinose, glucose, mannose, galactose, and fructose. Non-exclusive examples of disaccharides include sucrose, maltose, lactose, lactulose, and trehalose. Other “sugars,” “sugar groups,” or “sugar residues” include, but are not limited to, polysaccharides and / or oligosaccharides, including amylose, amylopectin, glycogen, inulin, and cellulose. In some cases, the “sugar,” “sugar group,” or “sugar residue” is an amino sugar. In some cases, the “sugar,” “sugar group,” or “sugar residue” is a glucamine residue (1-amino-1-deoxy-D-glucitol) (i.e., glucamide) that attaches to the remainder of the molecule via an amino group, forming an amide bond with the remainder of the molecule.

[0105] Certain groups, subgroups, substituents, and atoms are represented, for example, by a wavy line crossing the bond(s)(single or multiple) to indicate the atom to which that group, subgroup, substituent, or atom is bonded. For example, a phenyl group is represented as a propyl group as shown below: [ka] It has the following structure: [ka]

[0106] As used herein, “binding agent” refers to any molecule (e.g., an antibody) that is capable of binding with specificity to a given binding partner (e.g., an antigen).

[0107] As used herein, the term “amino acid” refers to an organic compound that contains an amino group (-NH2) and a carboxyl group (-COOH) along with a side chain (R group) specific to each amino acid. Amino acids can be either proteinogenic or non-proteinogenic. “Protogenic” means that an amino acid is one of the 20 naturally occurring amino acids found in proteins. Examples of proteinogenic amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. “Non-proteinogenic” means that the amino acid is not naturally present in proteins or is not directly produced by cellular mechanisms (e.g., it is a product of post-translational modification). Non-extendable examples of non-proteinogenic amino acids include gamma-aminobutyric acid (GABA), taurine (2-aminoethanesulfonic acid), theanine (L-γ-glutamylethylamide), hydroxyproline, beta-alanine, ornithine, and citrulline.

[0108] As used herein, “peptide” is defined in its broadest sense across its various grammatical forms and refers to a compound of two or more subunit amino acids, amino acid analogs, or other peptide mimetic compounds. The subunits may be linked by peptide bonds or by other bonds (e.g., esters, ethers, etc.). As used herein, the term “amino acid” refers to natural and / or unnatural or synthetic amino acids (including glycine and both D and L optical isomers), as well as amino acid analogs and peptide mimetic compounds. When the peptide chain is short (e.g., two, three, or more amino acids), it is generally called an oligopeptide. When the peptide chain is longer, the peptide is usually called a polypeptide or protein. Full-length proteins, analogs, variants, and fragments of these are included in this definition. The term also includes post-expression modifications of polypeptides, such as glycosylation, acetylation, and phosphorylation. Furthermore, because ionizable amino and carboxyl groups are present in the molecule, certain peptides can be obtained as acidic or basic salts, or in a neutral form. Peptides can be obtained directly from source organisms, or they can be produced through recombinant or synthetic means.

[0109] The amino acid sequences of antibodies can be numbered using any known numbering scheme, including those described in Kabat et al., ("Kabat" numbering scheme); Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 ("Chothia" numbering scheme); MacCallum et al., 1996, J. Mol. Biol. 262:732-745 ("Contact" numbering scheme); Lefranc et al., Dev. Comp. Immunol., 2003, 27:55-77 ("IMGT" numbering scheme); and Honegge and Pluckthun, J. Mol. Biol., 2001, 309:657-70 ("AHo" numbering scheme). Unless otherwise specified, the numbering scheme used herein is the Kabat numbering scheme. However, the selection of a numbering scheme is not intended to imply sequence differences where they do not exist, and those skilled in the art can easily determine the sequence location by examining the amino acid sequences of one or more antibodies. Unless otherwise stated, the "EU numbering scheme" is commonly used when referring to residues within the constant region of the antibody heavy chain (as reported, e.g., by Kabat et al. (cited above)).

[0110] As used herein, the term “cytotoxic activity” refers to activity that reduces or diminishes the cell viability of the cell line being tested.

[0111] In the claims below and the foregoing description, unless otherwise required by context due to express expression or necessary implication, the word “comprise,” or variations such as “comprises” or “comprising,” is used in a comprehensive sense, that is, it indicates the presence of the described features, but does not exclude the presence or addition of further features in various embodiments.

[0112] 7.2. PBD Compounds In this specification, compounds of formula (I): [ka] Or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof is described, where each of ring A and ring B is independently represented by the following formula: [ka] [ka] It is one of them, [ka] This indicates the linking point to the linker. The linker is -(CH2) r -,-(CH2) p -X-(CH2) q -, or -(CH2) p -CH=CH-(CH2) q -and, X is NR 6 , NHC(=O), C(=O)NH, O, SO2, substituted or unsubstituted divalent aryl rings, substituted or unsubstituted divalent heteroaryl rings, substituted or unsubstituted divalent heterocycles, or substituted or unsubstituted divalent cyclic rings, Ring C is a cyclopropyl ring or a cyclobutyl ring. -C(R 1 )- and -N(R 2 Each of the dotted line connections between ) and is independently a single or double bond. If the dotted line connection is a single connection, each R 1 Each R is independently either H or OH, and each R 2 H is independent of H, If the dotted line bond is a double bond, then each R 1 H is independent of each R 2 It is independently non-existent, R 3 and R 4 Each of these is independently H, NH2, and NR a Rb OH, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, and R a and R b These are H or C, respectively, independently. 1~4 It is alkyl, R 5 H, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, R 6 is H, or C 1~4 It is alkyl, Each of m, n, and o is independently either 1 or 2. Each of r, p, and q is an independent integer between 1 and 8. The sum of p and q is an integer between 1 and 8.

[0113] In some embodiments, X is NR 6 It is an NHC (=O), O, a substituted or unsubstituted aryl ring, a substituted or unsubstituted heteroaryl ring, a substituted or unsubstituted heteroring, or a substituted or unsubstituted cyclic ring.

[0114] In some embodiments, rings A and B have the same formula.

[0115] In some embodiments, ring A and ring B each have different formulas.

[0116] In some embodiments, the linker is -(CH2) r If this is the case, then rings A and B are not the same equation.

[0117] In some embodiments, the linker is -(CH2) r - If this is the case, then ring A is given by equation (IIa) and ring B is given by equation (IIb).

[0118] In some embodiments, the linker is -(CH2) p -X-(CH2) q -or-(CH2) p-CH=CH-(CH2) q -If ring A is of formula (IIa), m in ring A is 2, and ring B is one of formulas (IIa), (IIb), (IIc), (IId), (IIe), (IIf), and (IIg). In some embodiments, the linker is -(CH2) p -X-(CH2) q -or-(CH2) p -CH=CH-(CH2) q - If this is the case, then ring A is formula (IIa), m is 1 in ring A, and ring B is one of formulas (IIa), (IIb), (IIc), (IId), (IIe), (IIf), and (IIg). In some embodiments, formula (IIc) is formula (IIc3) or formula (IIc4) below: [ka]

[0119] In some embodiments, the linker is -(CH2) p -X-(CH2) q -or-(CH2) p -CH=CH-(CH2) q - If this is the case, then ring A is equation (IIa), and ring B is one of equations (IIa), (IIb), (IIc), (IId), (IIe), (IIf), or (IIg).

[0120] In some embodiments, the linker is -(CH2) r -,-(CH2) p -X-(CH2) q -, or -(CH2) p -CH=CH-(CH2) q - and X is O, NR 6 , NHC(=O), -(m-C6H4)-, [ka] or [ka] In some embodiments, R 6 It is either H or methyl.

[0121] In some embodiments, the linker is -(CH2) r - is

[0122] In some embodiments, r is 3 or 5.

[0123] In some embodiments, the linker is -(CH2) p -O-(CH2) q -or-(CH2) p -NH-(CH2) q - is

[0124] In some embodiments, the sum of p and q is 4.

[0125] In some embodiments, the linker is [ka] That is the case.

[0126] In some embodiments, the sum of p and q is 2.

[0127] In some embodiments, the linker [ka] That is the case.

[0128] In some embodiments, the sum of p and q is 2.

[0129] In some embodiments, the linker is -(CH2) p -CH=CH-(CH2) q - is

[0130] In some embodiments, the sum of p and q is 3.

[0131] In some embodiments, ring B is given by formula (IIa).

[0132] In some embodiments, m is 1.

[0133] In some embodiments, ring C is a cyclopropyl ring.

[0134] In some embodiments, the dotted line bond within ring B is a single bond, and R 1 is H or OH, and R 2 H is H.

[0135] In some embodiments, the dotted bond within ring B is a double bond, and R 1 H is R 2 It does not exist.

[0136] In some embodiments, ring A is given by formula (IIb).

[0137] In some embodiments, ring A is given by formula (IIb2): [ka]

[0138] In some embodiments, R 3 H is H.

[0139] In some embodiments, the dotted line bond within ring A is a single bond, and R 1 is H or OH, and R 2 H is H.

[0140] In some embodiments, the dotted bond within ring A is a double bond, and R 1 H is R 2 It does not exist.

[0141] In some embodiments, the compound is [ka] [ka] That is the case.

[0142] In some embodiments, ring A is given by formula (IIg).

[0143] In some embodiments, o is 2.

[0144] In some embodiments, the dotted line bond within ring A is a single bond, and R 1 is H or OH, and R 2 H is H.

[0145] In some embodiments, the dotted bond within ring A is a double bond, and R 1 H is R 2 It does not exist.

[0146] In some embodiments, the compound is [ka] That is the case.

[0147] In some embodiments, ring A is given by formula (IId).

[0148] In some embodiments, n is 1.

[0149] In some embodiments, the dotted line bond within ring A is a single bond, and R 1 is H or OH, and R 2 H is H.

[0150] In some embodiments, the dotted bond within ring A is a double bond, and R 1 H is R 2 It does not exist.

[0151] In some embodiments, the compound is [ka] That is the case.

[0152] In some embodiments, ring A is given by formula (IIc).

[0153] In some embodiments, ring A is given by formula (IIc2): [ka]

[0154] In some embodiments, R 4 It is CH3O-.

[0155] In some embodiments, the dotted line bond within ring A is a single bond, and R 1 is H or OH, and R 2 H is H.

[0156] In some embodiments, the dotted bond within ring A is a double bond, and R 1 H is R 2 It does not exist.

[0157] In some embodiments, the compound is [ka] That is the case.

[0158] In some embodiments, ring A is given by formula (IId).

[0159] In some embodiments, n is 2.

[0160] In some embodiments, the dotted line bond within ring A is a single bond, and R 1 is H or OH, and R 2 H is H.

[0161] In some embodiments, the dotted bond within ring A is a double bond, and R 1 H is R 2 It does not exist.

[0162] In some embodiments, the compound is [ka] That is the case.

[0163] In some embodiments, ring A is given by formula (IIe).

[0164] In some embodiments, R 5 It is methyl.

[0165] In some embodiments, the dotted line bond within ring A is a single bond, and R 1 is H or OH, and R 2 H is H.

[0166] In some embodiments, the dotted bond within ring A is a double bond, and R 1 H is R 2 It does not exist.

[0167] In some embodiments, the compound is [ka] That is the case.

[0168] In some embodiments, each of ring A and ring B independently contains formula (IIa).

[0169] In some embodiments, m is 1.

[0170] In some embodiments, ring C is a cyclopropyl ring.

[0171] In some embodiments, if the dotted line bond in ring A is a single bond, then R 1 is H or OH, and R 2 H is H.

[0172] In some embodiments, if the dotted bond in ring A is a double bond, then R 1 H is R 2It does not exist.

[0173] In some embodiments, if the dotted bond in ring B is a single bond, then R 1 is H or OH, and R 2 H is H.

[0174] In some embodiments, if the dotted bond in ring B is a double bond, then R 1 H is R 2 It does not exist.

[0175] In some embodiments, the compound is [ka] [ka] That is the case.

[0176] 7.3. Linker-Payload Compounds Compounds, or pharmaceutically acceptable salts, tautomers, solvates, or stereoisomers thereof, comprising a covalent linker bound to a residue of at least one PBD compound disclosed herein, are also disclosed herein. The covalent linker (or "Ab linker") can be bound to a binder to form a conjugate, for example, as described below.

[0177] In some embodiments, the compound is given by the following formula: [ka] Having one of the following, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, Each of rings A and B is independently one of the following equations: [ka] [ka] [ka] This indicates the linking point to the linker or Ab linker. The linker is -(CH2) r -,-(CH2) p -X-(CH2) q -, or -(CH2) p -CH=CH-(CH2) q -and, X is NR 6 , NHC(=O), C(=O)NH, O, SO2, substituted or unsubstituted aryl rings, substituted or unsubstituted heteroaryl rings, substituted or unsubstituted heterocycles, or substituted or unsubstituted cyclic rings, Ring C is a cyclopropyl ring or a cyclobutyl ring. -C(R 1 )- and -N(R 2 )- Dotted line connection between [ka] Each of these is independently a single bond or a double bond, If the dotted line connection is a single connection, each R 1 Each R is independently either H or OH, and each R 2 H is H, If the dotted line bond is a double bond, then each R 1 H is independent of each R 2 It does not exist, R 3 and R 4 Each of these is independently H, NH2, and NR a R b OH, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, R a and R b These are H or C, respectively, independently. 1~4 It is alkyl, R 5 H, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, R6 is H, or C 1~4 It is alkyl, Each of m, n, and o is independently either 1 or 2. Each of r, p, and q is an independent integer between 1 and 8. The sum of p and q is an integer between 1 and 8. An Ab linker is a compound that can link ring A or ring B to a binder.

[0178] In some embodiments, the Ab linker has the following formula: [ka] During the ceremony, [ka] This indicates a bond point to ring A or ring B.

[0179] In some embodiments, the compound has the following formula: [ka]

[0180] 7.4. Conjugate Also disclosed herein are conjugates, or pharmaceutically acceptable salts, tautomers, solvates, or stereoisomers thereof, comprising a protein that binds to a residue of at least one PBD compound disclosed herein via a covalent linker. In some embodiments, the protein is a conjugate such as an antibody or its antigen-binding fragment. The conjugate may be an antibody-drug conjugate (ADC).

[0181] In some embodiments, the protein is directly bound to a covalent linker described herein (e.g., an Ab linker). In such cases, the binder is located one binding site away from the covalent linker. Alternatively, the covalent linker may be directly bound to the payload residue such that the covalent linker is located one binding site away from the payload residue. The payload may be any PBD compound described herein.

[0182] In some embodiments, the conjugate is expressed by the following formula: [ka] Having one of the following, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, Each of rings A and B is independently one of the following equations: [ka] [ka] [ka] This indicates the linking point to the linker or Ab linker. The linker is -(CH2) r -,-(CH2) p -X-(CH2) q -, or -(CH2) p -CH=CH-(CH2) q -and, X is NR 6 , NHC(=O), C(=O)NH, O, SO2, substituted or unsubstituted aryl rings, substituted or unsubstituted heteroaryl rings, substituted or unsubstituted heterocycles, or substituted or unsubstituted cyclic rings, Ring C is a cyclopropyl ring or a cyclobutyl ring. -C(R 1 )- and -N(R 2Each of the dotted line connections between ) and is independently a single or double bond. If the dotted line connection is a single connection, each R 1 Each R is independently either H or OH, and each R 2 H is independent of H, If the dotted line bond is a double bond, then each R 1 H is independent of each R 2 It is independently non-existent, R 3 and R 4 Each of these is independently H, NH2, and NR a R b OH, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, and R a and R b These are H or C, respectively, independently. 1~4 It is alkyl, R 5 H, C 1~4 Alkyl, C 1~4 It is an alkoxyl or aryl, R 6 is H, or C 1~4 It is alkyl, Each of m, n, and o is independently either 1 or 2. Each of r, p, and q is an independent integer between 1 and 8. The sum of p and q is an integer between 1 and 8.

[0183] In some embodiments, the Ab linker has the following formula: [ka] [ka] This indicates the connection point to Ab, [ka] This indicates a bond point to ring A or ring B.

[0184] In some embodiments, the Ab linker has the following formula: [ka] [ka] This indicates the connection point to Ab, [ka] This indicates a bond point to ring A or ring B.

[0185] In some embodiments, the conjugate is expressed by the following formula: [ka] The formula comprises, where Ab is a binder selected from a humanized antibody, a chimeric antibody, a human antibody, or an antigen-binding fragment thereof, and the subscript x is 1 to 15. In some embodiments, x is approximately 2.

[0186] In some embodiments, the ADC is given by the following formula: [ka] The formula comprises a binding agent selected from a humanized antibody, a chimeric antibody, a human antibody, or an antigen-binding fragment thereof.

[0187] 7.5. Method or process for creating a conjugate A method for preparing a conjugate is provided herein by contacting a binder (BA) with a linker-payload compound under conditions suitable for bond formation between the binder and the linker-payload compound. The reaction conditions can be any suitable reaction conditions known in the art. The binder may be an antibody, and the binding may form an antibody-drug conjugate.

[0188] Examples of such reactions are shown in the following examples.

[0189] In some embodiments, the method for preparing the conjugate includes treating or contacting a compound with a binder under coupling conditions. The compound may include a reactive linker bound to at least one payload. This compound may be any linker compound or platform compound disclosed herein.

[0190] 7.6. Pharmaceutical Compositions This specification also provides compositions (including pharmaceutical compositions) containing the ADC described herein. In some embodiments, the composition (e.g., a pharmaceutical composition) further comprises pharmaceutically acceptable excipients.

[0191] Pharmaceutical compositions according to this disclosure can be prepared in the form of lyophilized formulations or aqueous solutions by mixing an antibody-drug conjugate of a desired degree of purity with one or more optionally selected pharmaceutically acceptable carriers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). Generally, pharmaceutically acceptable carriers are non-toxic to the recipient at the doses and concentrations used, and such carriers include, but are not limited to, buffers, e.g., phosphates, citrates, and other organic acids; antioxidants (including ascorbic acid and methionine); preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, or benzyl alcohol; alkylparabens, e.g., methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); and low molecular weight (about 1 Polypeptides (less than 0 residues); proteins, e.g., serum albumin, gelatin, or immunoglobulin; hydrophilic polymers, e.g., polyvinylpyrrolidone; amino acids, e.g., glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates (including glucose, mannose, or dextrin); chelating agents, e.g., EDTA; sugars (e.g., sucrose, mannitol, trehalose, or sorbitol); salt-forming counterions, e.g., sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants, e.g., polyethylene glycol (PEG). Exemplary pharmaceutically acceptable carriers as used herein include interstitial drug dispersants such as soluble neutral active hyaluronidase glycoproteins (sHASEGP), e.g., human soluble PH-20 hyaluronidase glycoproteins such as rHuPH20 (HYLENEX®, Baxter International, Inc.). Certain exemplary sHASEGPs and uses, including rHuPH20, are described in U.S. Patents US7,871,607 and US2006 / 0104968.In one embodiment, sHASEGP is combined with one or more glycosaminoglycans (e.g., chondroitinase).

[0192] Examples of lyophilized formulations are described in U.S. Patent No. 6,267,958. Aqueous formulations include those described in U.S. Patent No. 6,171,586 and WO2006 / 044908, the latter of which includes histidine-acetate buffer.

[0193] 7.7.How to use In some embodiments, methods for treating a disease or disorder (e.g., cancer) in a person in need (e.g., a patient) are described herein, which include administering a therapeutically effective dose of a conjugate disclosed herein to the patient.

[0194] The conjugates disclosed herein may be administered by any preferred means, including parenteral, intrapulmonary, intranasal, and, if desired for local treatment, intrafocal administration. Parenteral administration may include intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration. Administration may be carried out by any preferred route, for example, by injection (e.g., intravenous or subcutaneous injection), depending in part whether the administration is short-term or chronic. Various administration schedules, including but not limited to single or multiple doses, bolus administration, and pulse infusion, are contemplated herein.

[0195] The conjugates of this disclosure may be formulated, administered, and given in a manner consistent with good medical practice. Factors to be considered in this regard include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of drug delivery, the method of administration, the administration plan, and other factors known to the healthcare professional. [Examples]

[0196] 8. Examples The following examples are illustrative and should not be considered limiting in any way. Unless otherwise specified, the experimental methods in the following examples are conventional. Unless otherwise specified, reagents and materials are commercially available. All solvents and chemicals used are analytical grade or of chemical purity. Solvents were redistilled before use. Anhydrous solvents were prepared according to standard or reference methods. Silica gel for column chromatography (100-200 mesh) and silica gel for thin-layer chromatography (TLC) (GF254) are commercially available from Tsingdao Haiyang Chemical Co., Ltd. or Yantai Chemical Co., Ltd. in China, and both were eluted with petroleum ether (60-90°C) / ethyl acetate (v / v) and visualized with iodine or molybd phosphoric acid solution in ethanol unless otherwise specified. All extraction solvents were dried with anhydrous Na2SO4 unless otherwise specified. 1 ¹H NMR spectra were recorded using a Bruck-400, Varian 400MR nuclear magnetic resonance spectrometer (using TMS (tetramethylsilane) as an internal standard). Coupling constants are shown in Hertz. Peaks are reported as singlelines (s), doublelines (d), triplelines (t), quadruplines (q), quintuplines (p), hexatlines (h), heptuplines (hept), multilines (m), or combinations thereof. br stands for broad. LC / MS data were recorded using an Agilent 1100, 1200 high-performance liquid chromatography-ion trap mass spectrometer (LC-MSD trap) equipped with a diode array detector (DAD) detecting at 214 nm and 254 nm and an ion trap (ESI source). All compound names except for reagents were generated using ChemDraw® 18.0.

[0197] For the sake of brevity, this specification uses certain abbreviations. One example is the one-letter abbreviations for amino acids. The amino acids and their corresponding three-letter and one-letter abbreviations are as follows: [Table 11]

[0198] In the following examples, the following abbreviations will be used. [Table 12]

[0199] UPLC analysis method Method A: Mobile phase A: 0.1% FA in water, B: MeCN, Gradient: 10% B maintained for 0.2 minutes, 10%~95% B for 5.8 minutes, 95% B maintained for 0.5 minutes, Flow rate: 0.6 mL / min, Column: ACQUITY UPLC(registered trademark) BEH C18 1.7 μm.

[0200] Method B: Mobile phase A: 0.1% FA in water, B: MeCN, Gradient: 10% B maintained for 0.5 minutes, 10%~90% B for 2.5 minutes, 90% B maintained for 0.2 minutes, Flow rate: 0.6 mL / min, Column: ACQUITY UPLC(registered trademark) BEH C18 1.7 μm.

[0201] Method C: Mobile phase A: 0.1% FA in water, B: MeCN; Gradient: 10% B maintained for 0.2 minutes, 10%~90% B for 1.3 minutes, 90% B maintained for 0.3 minutes; Flow rate: 0.6 mL / min; Column: ACQUITY UPLC(registered trademark) BEH C18 1.7 μm.

[0202] Example 1-1 [ka] Step 1: (S)-3-hydroxy-2-methoxy-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1-1b)

[0203] Compound 1-1b was synthesized according to the procedure described in Bioorg Med Chem Lett. 2019 Sep 1;29(17):2455-2458.

[0204] Step 2: (S)-3-((5-iodopentyl)oxy)-2-methoxy-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1-1c)

[0205] 1-1b (50 mg, 0.16 mmol) and 1,5-diiodopentane (0.12 mL, 0.81 mmol) were dissolved in dry DMF (0.5 mL). The solution was cooled to 0°C, and K2CO3 (45 mg, 0.32 mmol) was added all at once. The mixture was warmed to room temperature and stirred at room temperature for 6 hours. Then, ethyl acetate (5 mL) was added, and the diluted organic phase was washed with H2O (10 mL) and brine (10 mL). The organic phase was dried over Na2SO4 and concentrated to obtain the crude product, which was purified by silica column gel chromatography (eluent: hexane / ethyl acetate = 100 / 0~25 / 75) to obtain 1-1c as a pale yellow solid (49 mg, yield 60%). MS(ESI) m / z: 505.3[M+H] + ; 1 H NMR(400MHz,CDCl3)δ7.53(s,1H),7.49(d,J=5.3Hz,1H),7.39-7.34(m,2H),7.31(dd,J=3.8,1.6Hz,2H),6.80(s,1H),5.01(d,J=15. 6Hz,1H),4.56(d,J=15.5Hz,1H),4.16-4.01(m,3H),3.95(s,3H),3.33-3.09(m,4H),1.95-1.84(m,4H),1.59(tt,J=9.8,6.1Hz,2H).

[0206] Step 3: Allyl(11S,11aS)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-8-((5-(((S)-2-methoxy-14-oxo-6a,7,12,14-tetrahydrobenzo[5,6][1,4]diazepine[1,2-b]isoquinoline-3-yl)oxy)pentyl)oxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-1d)

[0207] K2CO3 (12.5 mg, 0.09 mmol) was added to solutions of 1-1c (50 mg, 0.098 mmol) and 1-7 g (40 mg, 0.082 mmol) in 0.5 mL of DMF. The mixture was stirred at room temperature for 3 hours. LC-MS showed complete consumption of 1-7 g. Subsequently, siRNA (5 mL) was added, and the diluted organic phase was washed with H2O (10 mL) and brine (10 mL). The organic phase was dried over Na2SO4 and concentrated to obtain the crude product, which was purified by silica column gel chromatography (eluent: hexane / siRNA = 100 / 0~0 / 100) to obtain 1-1d as a white solid (60 mg, yield 70.6%). MS(ESI) m / z: 865.5[M+H] + .

[0208] Step 4: Allyl(11S,11aS)-11-hydroxy-7-methoxy-8-((5-(((S)-2-methoxy-14-oxo-6a,7,12,14-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-3-yl)oxy)pentyl)oxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-1e)

[0209] To a solution of 1-1d (60 mg, 0.07 mmol) in dry THF (1 mL), AcOH (24 μL) was added, followed by dropwise addition of TBAF (350 μL, 0.35 mmol, 1 M in THF). The mixture was stirred at room temperature for 6 hours. After checking for completeness by LC-MS, the mixture was quenched with saturated NaHCO3. The organic phase was extracted with  (5 mL x 3) and washed with H2O (10 mL) and brine (10 mL). The organic phase was dried over Na2SO4 and concentrated to obtain the crude product, which was purified by silica column gel chromatography (eluate: CH2Cl2 / MeOH = 100 / 0~5 / 95) to obtain 1-1e as a white solid (50 mg, yield 96%). MS(ESI) m / z: 751.5 [M+H] + .

[0210] Step 5: (S)-2-Methoxy-3-((5-((((S)-7-Methoxy-5-oxo-5,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1-1)

[0211] Pd(PPh3)4 (2 mg, catalytic amount) was added to a solution of 1-1e (50 mg, 0.067 mmol) in CH2Cl2 (1 mL) and pyrrolidine (14 μL, 0.17 mmol). The reaction mixture was stirred under N2 at room temperature for 0.5 hours. The reaction product was diluted with CH2Cl2 (5 mL) and washed with saturated NH4Cl and brine. The organic phase was dried over Na2SO4. The organic phase was concentrated and purified by silica column gel chromatography (eluate: CH2Cl2 / MeOH = 100 / 0~5 / 95) to obtain 1-1 as a white solid (43 mg, yield 99%). MS(ESI) m / z: 649.4 [M+H] + ; 1 H NMR(400MHz,CDCl3)δ7.79(d,J=4.4Hz,1H),7.52(d,J=7.3Hz,2H),7.48(d,J=5.2Hz,1H),7.40-7.29(m,4H),6. 80(d,J=4.4Hz,2H),5.01(d,J=15.5Hz,1H),4.56(d,J=15.5Hz,1H),4.19-4.02(m,4H),3.94(d,J=2.9Hz,7H),3. 87(ddd,J=7.8,4.5,2.7Hz,1H),3.68(d,J=11.7Hz,1H),3.55-3.47(m,1H),3.26(d,J=5.5Hz,1H),3.16(dd,J=15 .4,4.2Hz,1H),2.52(dd,J=13.0,8.1Hz,1H),2.08-1.90(m,6H),1.68(td,J=8.6,5.9Hz,2H),0.81-0.68(m,4H).

[0212] Examples 1-2 [ka] Step 1: (S)-3-(benzyloxy)-2-methoxy-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(5H)-one(1-2a)

[0213] NaBH(OAc)3 (531.9 mg, 2.5 mmol) was added at 0°C to a solution of 1-1a (500 mg, 1.255 mmol) in CH2Cl2 (6 mL). The reaction mixture was then warmed to room temperature and stirred under N2 at room temperature for 2 hours. The reaction mixture was then quenched with saturated NaHCO3, washed with H2O and brine, and dried over Na2SO4. The organic phase was filtered and concentrated to obtain crude product 1-2a as a white solid (485 mg, 97% yield), which was used directly in the next step without further purification. MS(ESI)m / z:401.3[M+H] + .

[0214] Step 2: Allyl(S)-3-(benzyloxy)-2-methoxy-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(1-2b)

[0215] Alloc-Cl (0.14 mL, 1.33 mmol) was added dropwise to a solution of 1-2a (485 mg, 1.2 mmol) and pyridine (0.36 mL, 2.9 mmol) in CH2Cl2 (2 mL) at 0°C. The reaction mixture was stirred under N2 at 0°C for 15 minutes. The reaction mixture was diluted with 10 mL of CH2Cl2, washed with 0.1 N citric acid (10 mL), H2O (10 mL), and brine (10 mL), and dried over Na2SO4. The organic phase was filtered, and the filtrate was concentrated. The crude product was purified by silica column gel chromatography (eluate: hexane / Â=100 / 0~50 / 50) to obtain 1-2b as a white solid (480 mg, yield 82%). MS(ESI) m / z: 485.5[M+H] + .

[0216] Step 3: Allyl(S)-3-hydroxy-2-methoxy-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(1-2c)

[0217] MeSO3H (0.64 mL, 9.9 mmol) was added dropwise to a solution of 1-2b (484.5 mg, 0.99 mmol) in CH2Cl2 (3 mL) at 0°C. The reaction mixture was stirred at 0°C for 10 minutes, then warmed to room temperature and stirred under N2 for 2 hours. The mixture was quenched with saturated NaHCO3. The organic phase was extracted with CH2Cl2 (5 mL x 3) and washed with H2O (10 mL) and brine (10 mL). The organic phase was dried over Na2SO4, filtered, and the filtrate was concentrated to obtain the crude product. This was purified by silica column gel chromatography (eluent: hexane / Â5 = 100 / 0~25 / 75) to obtain 1-2c as a white solid (332 mg, yield 84.3%). MS(ESI) m / z: 395.4 [M+H] + .

[0218] 1 H NMR(400MHz,CDCl3)δ7.28(d,J=3.3Hz,3H),7.21(s,2H),6.75(s,1H),5.88(s,1H),5 .76(ddd,J=17.3,10.5,5.2Hz,1H),5.10(dd,J=10.3,1.4Hz,2H),4.79-4.65(m,2H), 4.57(dd,J=13.9,5.2Hz,1H),4.40(d,J=13.3Hz,1H),4.05-3.95(m,2H),3.94(s,3H) ,3.37(d,J=11.4Hz,1H),3.12(dd,J=15.2,5.6Hz,1H),2.77(dd,J=15.2,4.1Hz,1H).

[0219] Step 4: Allyl(S)-3-((5-iodopentyl)oxy)-2-methoxy-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(1-2d)

[0220] 1-2d was prepared according to the procedure described in step 2 of Example 1-1 to obtain a white solid (138 mg, yield 92.2%). MS(ESI)m / z: 591.3[M+H] + .

[0221] 1 H NMR(400MHz,CDCl3)δ7.35(d,J=3.5Hz,3H),7.28(d,J=1.9Hz,2H),6.72(s,1H),5.84(ddt,J=16.3 ,10.8,5.2Hz,1H),5.25-5.08(m,2H),4.80(q,J=15.9Hz,1H),4.67(dd,J=13.7,5.3Hz,1H),4.48-4 .40(m,1H),4.14-4.00(m,4H),3.98(s,3H),3.46(d,J=10.7Hz,1H),3.29(t,J=7.0Hz,2H),3.20(d d,J=15.2,5.4Hz,1H),2.85(d,J=15.2Hz,1H),1.95(dq,J=13.4,7.0Hz,4H),1.66(q,J=8.0Hz,4H).

[0222] Step 5: Allyl(S)-3-((5-(((11S,11aS)-10-((allyloxy)carbonyl)-11-((tert-butyldimethylsilyl)oxy)-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)2-methoxy-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(1-2e)

[0223] 1-2e was prepared according to the procedure described in step 3 of Example 1-1 to obtain a colorless oil (70 mg, yield 85.3%). MS(ESI)m / z:973.6[M+Na] + .

[0224] Step 6: Allyl(S)-3-((5-(((11S,11aS)-10-((allyloxy)carbonyl)-11-hydroxy-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)2-methoxy-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(1-2f)

[0225] 1-2f was prepared according to the procedure described in step 4 of Example 1-1 to obtain a white solid (54 mg, yield 87.7%). MS(ESI)m / z:837.5[M+H] + .

[0226] Step 7: (S)-2-Methoxy-3-((5-(((S)-7-Methoxy-5-oxo-5,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(5H)-one(1-2)

[0227] 1-2 was prepared according to the procedure described in step 5 of Example 1-1 to obtain a white solid (40 mg, yield 95.2%). MS(ESI)m / z:651.4[M+H] + .

[0228] 1H NMR(400MHz,CDCl3)δ7.79(d,J=4.4Hz,1H),7.49(s,1H),7.35(s,1H),7.27(d,J=6.9Hz,4H),6.79(s,1H), 6.27(s,1H),4.87(d,J=15.7Hz,1H),4.73(d,J=15.7Hz,1H),4.23-3.97(m,5H),3.95(s,3H),3.85(s,3H),3 .67(d,J=11.7Hz,1H),3.53-3.41(m,2H),3.22(dd,J=12.1,9.5Hz,1H),3.11(dd,J=15.1,5.8Hz,1H),2.81( dd,J=15.2,5.3Hz,1H),2.51(dd,J=13.1,8.1Hz,1H),1.96(dq,J=24.2,9.3,8.1Hz,6H),1.72-1.62(m,2H).

[0229] Examples 1-3 and 1-4 [ka] Step 1: (S)-2-Methoxy-3-((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)-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1-3) and (S)-2-methoxy-3-((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)-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(5H)-one(1-4)

[0230] NaBH(OAc)3 (3.4 mg, 1.05 mmol) was added at 0°C to a solution of 1-1 (10 mg, 0.015 mmol) in CH2Cl2 (0.5 mL). The reaction mixture was then warmed to room temperature and stirred under N2 at room temperature for 0.5 hours. The reaction mixture was quenched with saturated NaHCO3, washed with H2O and brine, and dried over Na2SO4. The organic phase was filtered and concentrated to obtain the crude product, which was purified by preparative HPLC (0.01% FA in H2O) to obtain 1-3 (3 mg, yield 30%) as a white solid (retention time = 4.9 min) and 1-4 (2.7 mg, yield 27%) as a white solid (retention time = 5.12 min).

[0231] 1-3 MS(ESI)m / z:651.4[M+H] + .

[0232] 1 H NMR(400MHz,CDCl3)δ7.71-7.59(m,1H),7.55-7.43(m,3H),7.39-7.27(m,4H),6.76(s,1H),6.18(s,1H) ,4.98(d,J=15.5Hz,1H),4.53(d,J=15.5Hz,1H),4.17-3.95(m,5H),3.92(d,J=5.1Hz,3H),3.82(s,3H), 3.65(d,J=12.0Hz,1H),3.58-3.39(m,3H),3.25(dd,J=15.4,5.5Hz,1H),3.14(dd,J=15.4,4.3Hz,1H),2 .02(dd,J=12.7,7.3Hz,1H),1.91(h,J=7.3Hz,4H),1.68(dq,J=31.0,7.8,7.0Hz,4H),0.75-0.50(m,4H).

[0233] 1-4 MS(ESI)m / z:653.6[M+H] + .

[0234] 1H NMR(400MHz,CDCl3)δ7.56(s,1H),7.37(s,1H),7.32-7.27(m,5H),7.22-7.13(m,1H),6.21(s,1H),6.08(s,1H),4.88(d,J =15.7Hz,1H),4.75(d,J=15.7Hz,1H),4.14(d,J=6.2Hz,1H),4.00(q,J=6.1Hz,5H),3.89-3.79(m,6H),3.71(d,J=12.0Hz, 1H),3.60-3.51(m,2H),3.50-3.39(m,2H),3.23(t,J=10.8Hz,1H),3.12(dd,J=15.2,5.8Hz,1H),2.83(dd,J=15.2,5.4Hz, 1H),2.02(t,J=10.3Hz,1H),1.91(p,J=6.9Hz,4H),1.78(dd,J=12.7,6.9Hz,1H),1.66(q,J=7.8Hz,2H),0.78-0.52(m,4H).

[0235] Examples 1-5 [ka] Step 1: (S)-3-(2-(2-bromoethoxy)ethoxy)-2-methoxy-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1-5a)

[0236] 1-5a was prepared according to the procedure described in step 2 of Example 1-1 to obtain a white solid (50 mg, yield 67.1%).

[0237] MS(ESI)m / z:461.2[M+H] + .

[0238] Step 2: Allyl(11S,11aS)-11-hydroxy-7-methoxy-8-(2-(2-(((S)-2-methoxy-14-oxo-6a,7,12,14-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-3-yl)oxy)ethoxy)ethoxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-5c)

[0239] 1-5c was prepared according to the procedure described in step 3 of Example 1-1 to obtain a white solid (51 mg, yield 60.3%).

[0240] MS(ESI)m / z:753.5[M+H] + .

[0241] Step 3: (S)-2-Methoxy-3-(2-(2-((((S)-7-Methoxy-5-oxo-5,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)ethoxy)ethoxy)-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1-5)

[0242] Samples 1-5 were prepared according to the procedure described in step 4 of Example 1-1 to obtain a white solid (25.5 mg, yield 59%).

[0243] MS(ESI)m / z:651.4[M+H] + .

[0244] 1H NMR(400MHz,CDCl3)δ7.78(d,J=4.5Hz,1H),7.51(d,J=7.4Hz,2H),7.47(dd,J=5.3,3.3Hz,1H),7.40-7.30(m,4H) ,6.83(dd,J=7.3,4.5Hz,2H),5.00(d,J=15.5Hz,1H),4.56(d,J=15.5Hz,1H),4.37-4.16(m,4H),4.01(t,J=5.0Hz, 4H),3.96-3.90(m,6H),3.88-3.81(m,2H),3.67(d,J=11.7Hz,1H),3.49(d,J=11.7Hz,1H),3.27(dd,J=15.4,5.5Hz ,1H),3.15(dd,J=15.4,4.2Hz,1H),2.51(dd,J=13.1,8.1Hz,1H),1.99(dd,J=13.2,2.8Hz,1H),0.81-0.66(m,4H).

[0245] Examples 1-6 [ka] Step 1: 1-(tert-butyl)2-methyl(S)-4-methylenepiperidine-1,2-dicarboxylate(1-6b)

[0246] In a solution of MePh3PBr (1.56 g, 4.27 mmol) in dry THF (20 mL), KO in dry THF (1 M, 4.7 mL, 4.7 mmol) was added under an N2 atmosphere at 0°C. t Potassium tert-butoxide (Bu) was added and the mixture was stirred at 0°C for 1 hour. To the reaction solution, a solution of 1-6a (1.0 g, 3.89 mmol) in dry THF (20 mL) was added under an N2 atmosphere at 0°C and the mixture was stirred for 1 hour. Water (5 mL) and saturated NH4Cl (10 mL) were added to the solution. The organic phase was separated, and the aqueous phase was extracted with RINKAN (20 mL x 3). The combined organic phase was concentrated and purified by flash column chromatography (petroleum ether / RINKAN = 90 / 10) to obtain the title compound 1-6b as a colorless oil (588 mg, yield 59.3%). MS(ESI) m / z: 156.1 [M+H-Boc] + .

[0247] 1 H NMR(400MHz,CDCl3)δ5.08-4.97(m,0.5H),4.87-4.78(m,0.5H),4.79(s,2H),4.20-3.97(m,1H),3. 71(s,3H),3.12-2.90(m,1H),2.81-2.67(m,1H),2.49-2.37(m,1H),2218-2.10(m,2H),1.47(s,9H).

[0248] Step 2: Benzyl 2-methyl(S)-4-methylenepiperidine-1,2-dicarboxylate(1-6c)

[0249] To a solution of 1-6b (585 mg, 2.29 mmol) in dry MeOH (3 mL), 3N HCl in MeOH (7 mL) was added under an N2 atmosphere at 0°C, and the mixture was stirred at room temperature for 2 hours. The solution was concentrated and dissolved in CH2Cl2 (10 mL). 4N HCl in  (10 mL) was added, and the mixture was stirred for 20 minutes. The solution was concentrated, and the residue was dissolved in CH2Cl2 (10 mL). To the solution, CbzCl (benzyl chloroformate, 0.40 mL, 2.75 mmol) and TEA (0.65 mL, 4.58 mmol) were added at 0°C, and the mixture was stirred at room temperature for 30 minutes. The solution was added to 0.5N HCl (3 mL) and water (3 mL), and extracted with CH2Cl2 (5 mL x 3). The organic phase was concentrated and purified by flash column chromatography (petroleum ether / siRNA = 90 / 10) to obtain the title compound 1-6c as a colorless oil (310 mg, yield 46.8%). MS(ESI)m / z: 312.2[M+Na] + .

[0250] 1 H NMR(400MHz,CDCl3)δ7.45-7.24(m,5H),5.17(s,2H),5.14-4.90(m,1H),4.81(s,2H),4.28-4.08(m,1H), 3.69(d,J=18.8Hz,3H),3.23-3.01(m,1H),2.77(t,J=15.7Hz,1H),2.50-2.39(m,1H),2.30-2.12(m,2H).

[0251] Step 3: 6-benzyl 5-methyl(S)-6-azaspiro[2.5]octane-5,6-dicarboxylate(1-6d)

[0252] To 5 mL of dry CH2Cl2, ZnEt2 (2 M in hexane, 2.1 mL, 4.2 mmol) was added under an N2 atmosphere at 0°C, and the mixture was stirred at 0°C for 10 minutes. Dry TFA (0.33 mL, 4.2 mmol) was slowly added, and the mixture was stirred at 0°C for 1 hour. Diiodomethane (0.35 mL, 4.2 mmol) was slowly added, and the mixture was stirred at 0°C for 1 hour. A solution of 1-6c (305 mg, 1.05 mmol) in 3 mL x 2 of dry CH2Cl2 was slowly added, and the mixture was stirred at 0°C for 30 minutes, followed by stirring at room temperature for 18 hours. This solution was filtered through Celite. The filtrate was washed with saturated NH4Cl (5 mL) and water (5 mL). The aqueous phase was extracted with CH2Cl2 (5 mL x 3). The organic phase was concentrated and purified by flash column chromatography (petroleum ether / siRNA = 90 / 10) to obtain compounds 1-6d as a pale yellow oil (195 mg, yield 61%). MS(ESI)m / z: 326.3[M+Na] + .

[0253] 1 H NMR(400MHz,CDCl3)δ7.45-7.25(m,5H),5.24-5.08(m,2H),5.04-4.84(m,1H),4.21-4.02(m,1H),3.72(d,J=6Hz,3H ),3.36-3.14(m,1H),2.24-2.12(m,1H),2.00-1.85(m,1H),1.62-1.54(m,1H),0.91-0.72(m,1H),0.42-0.22(m,4H).

[0254] Step 4: Benzyl(S)-5-(hydroxymethyl)-6-azaspiro[2.5]octane-6-carboxylate(1-6e)

[0255] To a solution of 1-6d (195 mg, 0.64 mmol) in dry THF (4 mL), LiBHEt3 (1 M in THF, 1.3 mL, 1.3 mmol) was added under an N2 atmosphere at 0°C, and the mixture was stirred at 0°C for 1 hour. Water (0.5 mL) was added to the solution, washed with brine (5 mL), and extracted with  (5 mL x 3). The organic phase was concentrated and purified by flash column chromatography (petroleum ether /  = 50 / 50) to obtain the title compound 1-6e as a colorless oil (155 mg, yield 87.6%). MS(ESI) m / z: 276.3 [M + H] + .

[0256] 1 H NMR(400MHz,CDCl3)δ7.40-7.29(m,5H),5.22-5.09(m,2H),4.53-4.39(m,1H),4.20-4.06(m,1H),4.02(dd,J=11.0,9.5Hz,1H),3.67(dd,J=11.1, 5.6Hz,1H),3.19-3.05(m,1H),2.08-1.97(m,1H),1.95-1.81(m,1H),1.0 8-0.97(m,1H),0.91-0.79(m,2H),0.47-0.38(m,1H),0.35-0.23(m,3H).

[0257] Step 5: (S)-(6-azaspiro[2.5]octan-5-yl)methanol(1-6f)

[0258] To a solution of 1-6e (155 mg, 0.56 mmol) in MeOH (2 mL), 7 M NH3 in MeOH (0.2 mL) and 10% wet Pd / C (16 mg) were added under an N2 atmosphere, and the mixture was stirred under an H2 atmosphere for 3 hours. The solution was filtered and concentrated to obtain the title compound 1-6f as a yellowish-white oil (82 mg, quantified), which was used directly in the next step without further purification. MS(ESI)m / z:142.1[M+H] + .

[0259] Step 6: (S)-(4-(benzyloxy)-5-methoxy-2-nitrophenyl)(5-(hydroxymethyl)-6-azaspiro[2.5]octan-6-yl)methanone (1-6h)

[0260] To a solution of 1-6 g (160 mg, 0.53 mmol) in CH2Cl2 (4 mL), oxalyl chloride (0.14 mL, 1.58 mmol) was added under an N2 atmosphere at 0°C, followed by the addition of one drop of DMF, and the mixture was stirred for 20 minutes. The solution became clear, and no gas elution occurred. The solution was concentrated to remove excess oxalyl chloride. The residue was dissolved in dry CH2Cl2 (2 mL) and added to a solution of 1-6f (80 mg, 0.53 mmol) and DIPEA (0.38 mL, 2.1 mmol) in dry CH2Cl2 (2 mL) at 0°C, and the mixture was stirred for 20 minutes. The solution was added to water (5 mL) and extracted with CH2Cl2 (5 mL x 3). The organic phase was concentrated and purified by flash column chromatography (petroleum ether / Â5 = 30 / 70) to obtain the title compound 1-6h as a grayish-white solid (178 mg, yield 79%). MS(ESI)m / z:427.4[M+H] + .

[0261] Step 7: (S)-(2-amino-4-(benzyloxy)-5-methoxyphenyl)(5-(hydroxymethyl)-6-azaspiro[2.5]octan-6-yl)methanone(1-6i)

[0262] To a solution of 1-6h (175 mg, 0.41 mmol) and NH4Cl (336 mg, 6.16 mmol) in MeOH / H2O (3 / 1 mL), Fe powder (117 mg, 2.05 mmol) was added under an N2 atmosphere and refluxed for 3 hours. This solution was filtered through elite. The filtrate was washed with brine (5 mL), extracted with SiO (5 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated to obtain the title compound 1-6i (162 mg, quantified), which was used directly in the next step without further purification. MS(ESI)m / z:397.3[M+H] + .

[0263] Step 8: Allyl(S)-(5-(benzyloxy)-2-(5-(hydroxymethyl)-6-azaspiro[2.5]octane-6-carbonyl)-4-methoxyphenyl)carbamate(1-6j)

[0264] To a solution of 1-6i (162 mg, 0.41 mmol) and pyridine (67 μL, 0.82 mmol) in dry CH2Cl2 (4 mL), AllocCl (53 μL, 0.49 mmol) was added under a N2 atmosphere at -10°C, and the mixture was stirred for 20 minutes. The solution was added to water (2 mL) and 0.5 N HCl (2 mL) and extracted with CH2Cl2 (5 mL x 3). The organic phase was concentrated and purified by flash column chromatography (petroleum ether / Â=25 / 75) to obtain the title compound 1-6j as a grayish-white solid (157 mg, yield 80%). MS(ESI) m / z: 481.4 [M+H] + .

[0265] Step 9: Allyl(6aS)-3-(benzyloxy)-6-hydroxy-2-methoxy-12-oxo-6a,7,9,10-tetrahydro-6H-spiro[benzo[e]pyrido[1,2-a][1,4]diazepine-8,1'-cyclopropane]-5(12H)-carboxylate(1-6k)

[0266] To a solution of 1-6j (155 mg, 0.32 mmol) in CH2Cl2 (3 mL), DMP (209 mg, 0.48 mmol) was added at 0°C and the mixture was stirred at room temperature for 30 minutes. Saturated Na2S2O3 (2 mL) and saturated NaHCO3 (2 mL) were added to the solution and extracted with CH2Cl2 (5 mL x 3). The organic phase was concentrated and purified by flash column chromatography (petroleum ether / Â=50 / 50) to obtain the title compound 1-6k as a white solid (130 mg, yield 84%). MS(ESI)m / z:479.4[M+H] + .

[0267] Step 10: Allyl(6aS)-3,6-dihydroxy-2-methoxy-12-oxo-6a,7,9,10-tetrahydro-6H-spiro[benzo[e]pyrido[1,2-a][1,4]diazepine-8,1'-cyclopropane]-5(12H)-carboxylate(1-6l)

[0268] To a solution of 1-6k (75 mg, 0.16 mmol) in CH2Cl2 (2.5 mL), MsOH (104 μL, 1.56 mmol) was added under an N2 atmosphere at 0°C, and the mixture was stirred at room temperature for 5 hours. The solution was added to brine (3 mL) and extracted with CH2Cl2 / MeOH (10:1, 5.5 mL x 3). The organic phase was concentrated and purified by flash column chromatography (petroleum ether / Â=50 / 50) to obtain 1-6l of the title compound as a white solid (20 mg, yield 33%). MS(ESI)m / z:389.3[M+H] + .

[0269] 1 H NMR(400MHz,CDCl3)δ7.18(s,1H),6.73(s,1H),6.11(d,J=10.3Hz,1H),5.91(s,1H),5.90-5.75(m,1H), 5.28-5.10(m,2H),4.66(dd,J=13.0,5.1Hz,1H),4.57-4.46(m,1H),4.42(dt,J=13.4,4.5Hz,1H),3.96( s,3H),3.61-3.52(m,1H),3.45(brs,1H),3.23(ddd,J=13.5,11.5,4.1Hz,1H),2.02(dd,J=15.5,7.4Hz, 1H),1.91-1.80(m,1H),1.58(d,J=14.3Hz,1H),1.35-1.27(m,1H),0.62-0.46(m,2H),0.45-0.33(m,2H).

[0270] Step 11: Allyl(6aS)-3-((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]diazepine-8-yl)oxy)pentyl)oxy)-6-hydroxy-2-methoxy-12-oxo-6a,7,9,10-tetrahydro-6H-spiro[benzo[e]pyrido[1,2-a][1,4]diazepine-8,1'-cyclopropane]-5(12H)-carboxylate(1-6n)

[0271] To a solution of 1-6l (20 mg, 0.05 mmol) and 1-6m (31 mg, 0.05 mmol (synthesized according to the procedure described in US20200261594A1)) in dry DMF (1 mL), K2CO3 (8.7 mg, 0.06 mmol) was added under an N2 atmosphere, and the mixture was stirred at room temperature for 4 days. The solution was added to water (12 mL) and extracted with CH2Cl2 (5 mL x 4). The organic phase was concentrated and purified by flash column chromatography (petroleum ether / Â=25 / 75) to obtain the title compound 1-6n as a grayish-white solid (40 mg, yield 87%). MS(ESI)m / z:893.6[M+H] + .

[0272] Step 12: (S)-2-Methoxy-3-((5-(((S)-7-Methoxy-2-(4-Methoxyphenyl)-5-oxo-5,10,11,11a-Tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)-6a,7,9,10-tetrahydro-12H-spiro[benzo[e]pyrido[1,2-a][1,4]diazepine-8,1'-cyclopropane]-12-one(1-6)

[0273] To a solution of 1-6n (35 mg, 0.04 mmol) and Pd(PPh3)4 (1.2 mg, 0.001 mmol) in dry CH2Cl2 (1 mL), pyrrolidine (4 μL, 0.04 mmol) was added under an N2 atmosphere, and the mixture was stirred at room temperature for 30 minutes. The solution was concentrated and purified by flash column chromatography (petroleum ether / Â=0 / 100) to obtain the title compound 1-6 as a grayish-white solid (16 mg, yield 57.8%). MS(ESI)m / z:707.5[M+H] + .

[0274] 1 H NMR(400MHz,CDCl3)δ8.10-8.01(m,1H),7.54-7.47(m,2H),7.44(d,J=6.2Hz,1H),7.31(d,J=8.7Hz,2H),6.88(d,J=8.8H) z,2H),6.77(d,J=8.2Hz,1H),6.23-6.05(m,1H),4.37-4.21(m,2H),4.19-4.04(m,2H),4.00(t,J=6.6Hz,2H),3.95(d,J= 9.6Hz,3H),3.85(d,J=3.2Hz,3H),3.82(s,3H),3.64-3.50(m,2H),3.49-3.29(m,2H),2.73(dd,J=16.1,3.5Hz,1H),2.23 (dd,J=14.5,5.9Hz,1H),2.00-1.85(m,5H),1.76-1.59(m,4H),1.49-1.38(m,2H),0.72-0.57(m,2H),0.55-0.43(m,2H).

[0275] Examples 1-7 [ka] [ka] Step 1: Allyl(S)-(2-(6-(((tert-butyldimethylsilyl)oxy)methyl)-5-azaspiro[2,4]heptane-5-carbonyl)-4-methoxy-5-((triisopropylsilyl)oxy)phenyl)carbamate(1-7b)

[0276] Pyridine (433 μL) was added to a solution of 1-7a (1370 mg, 2.44 mmol) in dry CH2Cl2 at -5°C. Then, AllocCl (322 μL) was added to the mixture at -5°C, and the mixture was stirred at -5°C for 1 hour. Completion of the reaction was observed by TLC (petroleum ether / siRNA = 5:1). The mixture was diluted with CH2Cl2, washed with 5% citric acid, saturated NaHCO3, and brine, and dried over Na2SO4. The organic phase was concentrated to obtain the crude product, which was used directly in the next step. MS(ESI) m / z: 647.4[M+H] + .

[0277] Step 2: Allyl(S)-(2-(6-(hydroxymethyl)-5-azaspiro[2,4]heptane-5-carbonyl)-4-methoxy-5-((triisopropylsilyl)oxy)phenyl)carbamate(1-7c)

[0278] Para-toluenesulfonic acid hydrate (282 mg, 1.49 mmol) was added to a solution of 1-7b (1.60 g, 2.48 mmol) in THF (20 mL) and water (1 mL). The reaction mixture was stirred at 22°C for 1 hour. The completed reaction product was observed by TLC (petroleum ether / SiO7 = 5:1, 1:1), diluted with SiO7 (60 mL), and washed with water and brine. The organic phase was concentrated and purified by flash column chromatography to obtain 1.09 g of 1-7c (83% yield). MS(ESI) m / z: 533.3 [M+H] + .

[0279] Step 3: Allyl(11S,11aS)-11-hydroxy-7-methoxy-5-oxo-8-((triisopropylsilyl)oxy)-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-7d)

[0280] Anhydrous DMSO (436 μL, 6.14 mmol) was added dropwise to a solution of oxalyl chloride (260 μL, 3.07 mmol) in dry CH2Cl2 (20 mL) at -70°C. After 30 minutes, while maintaining the temperature at -70°C, a solution of 1-7c (1.09 g, 2.05 mmol) in dry CH2Cl2 (10 mL) was slowly added. After 40 minutes, triethylamine (1423 μL, dried over a 4 Å molecular sieve) was added dropwise, and the temperature was raised to -50°C in 1 hour. The reaction mixture was warmed to room temperature and stirred for 1 hour. Completion of the reaction was observed by TLC (petroleum ether / siRNA = 1:1, CH2Cl2 / siRNA = 10:1). The reaction mixture was washed with 5% citric acid aqueous solution (10V) to pH = 3. The organic phase was washed with saturated NaHCO3 aqueous solution and water, and dried over sodium sulfate. The organic phase was concentrated to obtain the crude product, which was purified by flash column chromatography (CH2Cl2 / Â=95 / 5) to obtain 1-7d (416 mg, yield 38%). MS(ESI)m / z:531.3[M+H] + .

[0281] Step 4: Allyl(11S,11aS)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-8-((triisopropylsilyl)oxy)-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-7e)

[0282] TBSOTf (0.54 mL, 2.35 mmol) was added to a mixture of 1-7d (416 mg, 0.78 mmol) and 2,6-lutidine (0.37 mL, 3.14 mmol) in dried CH2Cl2 (10 mL) at 0°C. The reaction mixture was stirred at 5°C for 30 minutes, followed by 25°C for 1 hour. Completion was observed by LC-MS. The reaction mixture was washed with saturated aqueous NaHCO3 and brine, dried over Na2SO4, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography (petroleum ether / Â=80 / 20) to obtain 1-7e (486 mg, 96% yield). MS(ESI) m / z: 645.5[M+H] + .

[0283] Step 5: Allyl(11S,11aS)-11-((tert-butyldimethylsilyl)oxy)-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)-carboxylate(1-7f)

[0284] Lithium acetate (50 mg, 0.73 mmol) was added to a solution of 1-7e (468 mg, 0.73 mmol) in wet DMF (10 mL, 49 / 1 DMF / water). The reaction was allowed to proceed at 25°C for 2 hours. Completion was observed by TLC (petroleum ether / siRNA = 2:1, 1:1). The mixture was diluted with siRNA and washed with 5% aqueous citric acid and brine. The organic phase was dried over Na2SO4 and concentrated to obtain the crude product, which was purified by flash column chromatography to obtain 1-7f (303 mg, yield 85%). MS(ESI) m / z: 489.4[M+H] + .

[0285] Step 6: Allyl(11S,11aS)-8-((5-bromopentyl)oxy)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate (1-7g)

[0286] To a solution of 1-7f (100 mg, 0.20 mmol) and 1,5-dibromopentane (0.42 mL, 3.07 mmol) in 2 mL of DMF, K2CO3 (34 mg, 0.25 mmol) was added at room temperature. The mixture was stirred at room temperature for 2 hours. LC-MS showed that the starting materials were completely consumed. The mixture was diluted with siRNA and washed with H2O and brine. The organic phase was concentrated and purified by flash column chromatography (petroleum ether / siRNA) to obtain 1-7 g (110 mg, yield 84%). MS(ESI)m / z:637.4.3[M+H] + .

[0287] Step 7: tert-butyl(S)-2-(hydroxymethyl)-4-methylenepiperidine-1-carboxylate (1-7h)

[0288] LiCl (180 mg, 4.23 mmol) in water (0.67 mL) was added to 1-6b (830 mg, 3.25 mmol) in dry THF (10 mL) at room temperature. Then, NaBH4 (160 mg, 4.23 mmol) was added to the mixture. The reaction mixture was stirred overnight at room temperature. 2N HCl (5V) was added to the mixture at 0°C, followed by saturated NaHCO3 being added to the mixture until the pH reached 7-8. The reaction mixture was extracted with HCl, washed with brine, dried over Na2SO4, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography (petroleum ether / HCl = 75 / 25) to obtain 1-7h (495 mg, yield 71%).

[0289] 1H NMR(400MHz,CDCl3)δ4.79(d,J=23.7Hz,2H),4.41(d,J=6.1Hz,1H),4.06(s,1H),3.67(dd,J=11.1,9.0Hz,1 H),3.57(dd,J=11.2,5.9Hz,1H),2.91(s,1H),2.37(dd,J=14.1,6.1Hz,1H),2.27-2.15(m,3H),1.48(s,9H).

[0290] Step 8: (S)-2-(hydroxymethyl)-4-methylenepiperidine-1-ium chloride (1-7i)

[0291] 4M HCl in 6mL of MeOH was added at 0°C to 1-7h (525 mg, 2.31 mmol) in 5mL of MeOH. The reaction mixture was stirred at room temperature for 2 hours. Completion was observed by TLC (petroleum ether / HCl = 2:1). The reaction mixture was washed with saturated NaHCO3 aqueous solution and water, dried over Na2SO4, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography to obtain 1-7i (425 mg). MS(ESI) m / z: 128.1 [M+H] + .

[0292] Step 9: (S)-(2-(hydroxymethyl)-4-methylenepiperidine-1-yl)(5-methoxy-2-nitro-4-((triisopropylsilyl)oxy)phenyl)methanone(1-7k)

[0293] Oxalyl chloride (603 μL, 7.07 mmol) was added dropwise to a stirred solution of 1-7i (870 mg, 2.36 mmol) in dry CH2Cl2 (8 mL), THF (8 mL), and DMF (4 μL) at 0°C under N2. The reaction mixture was warmed to room temperature and stirred for 1 hour. Completion was observed by TLC (petroleum ether / siRNA = 1:1). The mixture was concentrated to obtain the crude product, which was used directly in the next step. The crude product and 1-7j (423 mg, 2.59 mmol) were dissolved in CH2Cl2 (8 mL). The reaction mixture was cooled to 0°C and triethylamine (983 μL, 7.07 mmol) was added dropwise under N2. The mixture was then warmed to room temperature and stirred for 3 hours. The solution was concentrated, and the crude product was purified by flash column chromatography to obtain 1-7k (788 mg, 79% yield). MS(ESI)m / z:479.4[M+H] + .

[0294] Step 10: (S)-(2-amino-5-methoxy-4-((triisopropylsilyl)oxy)phenyl)(2-(hydroxymethyl)-4-methylenepiperidine-1-yl)methanone(1-7l)

[0295] Zinc powder (1.77 g, 27.08 mmol) was added at 0°C to a mixture of ethanol (4 mL), water (0.25 mL), and AcOH (0.25 mL). The reaction mixture was stirred at 5°C for 30 minutes. A solution of 1-7K (0.35 g, 0.73 mmol) in ethanol (2 mL) was added dropwise at 5°C. The reaction was allowed to proceed at 5°C for 30 minutes. The solid was removed by filtration. The filtrate was diluted with ethyl acetate and washed with water, saturated NaHCO3 aqueous solution, and brine. The organic phase was dried over sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure to obtain the product as brown oil (268 mg, yield 82%), which was used directly in the next step. MS(ESI)m / z:449.3[M+H] + .

[0296] Step 11: Allyl(S)-(2-(2-(hydroxymethyl)-4-methylenepiperidine-1-carbonyl)-4-methoxy-5-((triisopropylsilyl)oxy)phenyl)carbamate(1-7m)

[0297] Pyridine (104 μL) was added to a solution of 1-7 L (268 mg, 0.60 mmol) in dry CH2Cl2 at -5°C. Then, AllocCl (64 μL, 72.04 mmol) was added to the mixture at -5°C, and the mixture was stirred at -5°C for 0.5 hours. Completion of the reaction was observed by LC-MS. The mixture was diluted with CH2Cl2, washed with 5% citric acid, saturated NaHCO3, and brine, and dried over Na2SO4. The organic phase was concentrated to obtain the crude product, which was purified by flash column chromatography (petroleum ether / Â=65 / 35) to obtain 1-7 m (236 mg, yield 74%). MS(ESI) m / z: 533.4[M+H] + .

[0298] Step 12: Allyl(6S,6aS)-6-hydroxy-2-methoxy-8-methylene-12-oxo-3-((triisopropylsilyl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5(12H)-carboxylate(1-7n)

[0299] DMP (172 mg, 0.41 mmol) was slowly and gradually added to a solution of 1-7 m (206 mg, 0.39 mmol) in 5 mL of dry CH2Cl2 at 0°C. The reaction mixture was then warmed to room temperature and stirred for 2 hours. 0.5 equivalents of DMP were added to the reaction mixture in batches. After 9 hours, the starting material was consumed. The mixture was quenched with saturated Na2S2O3, followed by the addition of saturated NaHCO3 and water. The layers were separated, and the organic layer was washed with saturated Na2S2O3, saturated NaHCO3, and brine, and dried over Na2SO4. The crude product was purified by flash column chromatography (CH2Cl2 / siRNA = 93 / 7) to obtain 1-7 n (157 mg, 85% yield). MS(ESI) m / z: 531.3[M+H] + .

[0300] Step 13: Allyl(6S,6aS)-6-((tert-butyldimethylsilyl)oxy)-2-methoxy-8-methylene-12-oxo-3-((triisopropylsilyl)oxy)-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5(12H)-carboxylate(1-7o)

[0301] TBSOTf (0.23 mL, 1.0 mmol) was added to a mixture of 1-7n (177 mg, 0.33 mmol) and 2,6-lutidine (0.16 mL) in dried CH2Cl2 (5 mL) at 0°C. The reaction mixture was stirred at 5°C for 30 minutes, followed by 25°C for 1 hour. The reaction mixture was washed with saturated NaHCO3 aqueous solution and brine, dried over Na2SO4, and concentrated to obtain the crude product. The crude product was purified by flash column chromatography (CH2Cl2 / Â=98 / 2) to obtain 1-7o (135 mg, yield 63%). MS(ESI) m / z: 645.4[M+H] + .

[0302] Step 14: Allyl(6S,6aS)-6-((tert-butyldimethylsilyl)oxy)-3-hydroxy-2-methoxy-8-methylene-12-oxo-6,6a,7,8,9,10-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5(12H)-carboxylate(1-7p)

[0303] Lithium acetate (14 mg, 0.21 mmol) was added to a solution of 1-7o (135 mg, 0.21 mmol) in wet dimethylformamide (3 mL, 49 / 1 DMF / water). The reaction was allowed to proceed at 25°C for 2 hours. Completion was observed by TLC (petroleum ether / ethylacetate = 1:1). The mixture was diluted with ethylacetate and washed with 5% aqueous citric acid solution, saturated NaHCO3, and brine. The organic phase was dried over Na2SO4 and concentrated to obtain the crude product, which was purified by flash column chromatography to obtain 1-7p (100 mg, 98% yield). MS(ESI) m / z: 489.3 [M+H] + .

[0304] Step 15: Allyl(11S,11aS)-8-((5-(((6S,6aS)-5-((allyloxy)carbonyl)-6-((tert-butyldimethylsilyl)oxy)-2-methoxy-8-methylene-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-3-yl)oxy)pentyl)oxy)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-7q)

[0305] 1-7p (28 mg, 0.06 mmol) and K2CO3 (10 mg, 0.07 mmol) were added to a solution of 1-7 g (38 mg, 0.06 mmol) in 1 mL of DMF. The mixture was stirred overnight at room temperature. The product (petroleum ether / siRNA = 1:2) was detected by LC-MS. The mixture was diluted with siRNA and washed with water and brine. The organic phase was concentrated and purified by flash column chromatography (petroleum ether / siRNA = 27 / 73) to obtain 1-7q (48 mg, 84% yield). MS(ESI) m / z: 1045.7[M+H] + .

[0306] Step 16: Allyl(11S,11aS)-8-((5-(((6S,6aS)-5-((allyloxy)carbonyl)-6-hydroxy-2-methoxy-8-methylene-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-3-yl)oxy)pentyl)oxy)-11-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)-carboxylate(1-7r)

[0307] 1M TBAF (230 μL, 0.23 mmol) was added to a solution of 1-7q (48 mg, 0.05 mmol) in 3 mL of dry THF and AcOH (16 μL). The mixture was stirred at room temperature for 2 hours. After the reaction was complete by LC-MS, the mixture was diluted with SiO2 and washed with saturated NaHCO3 and brine. The organic phase was concentrated and purified by flash column chromatography to obtain 1-7r (21 mg, 56% yield). MS(ESI) m / z: 817.5[M+H] + .

[0308] Step 17: (S)-7-Methoxy-8-((5-((((S)-2-Methoxy-8-methylene-12-oxo-6a,7,8,9,10,12-Hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-3-yl)oxy)pentyl)oxy)-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one(1-7)

[0309] Pd(PPh3)4 (2.2 mg, 0.002 mmol) was added to a solution of 1-7r (15 mg, 0.02 mmol) in CH2Cl2 (0.5 mL) and pyrrolidine (8 μL, 0.09 mmol). The reaction mixture was stirred at room temperature for 0.5 hours. The reaction product was diluted with CH2Cl2 (10 mL) and washed with saturated NH4Cl and brine. The organic phase was concentrated and purified by preparative HPLC (0.01% FA in H2O) to obtain 1-7 (9 mg, 80% yield) as a white solid. MS(ESI) m / z: 613.4 [M+H] + .

[0310] 1 H NMR(400MHz,CDCl3)δ7.82(d,J=5.1Hz,1H),7.78(d,J=5.1Hz,1H),7.51(s,1H),7.47 s,1H),6.81(d,J=7.2Hz,2H),5.09(d,J=28.2Hz,2H),4.21-4.02(m,4H),4.03-3.74(m,11H),3.68(d,J=11.7Hz,1H),3.4 9(d,J=11.7Hz,1H),2.87-2.75(m,1H),2.70-2.64(m,2H),2.53(dd,J=12.9,8.1Hz,2H),1.97(m,6H),0.84-0.66(m,4H).

[0311] Examples 1-8 [ka] Step 1: diallyl 8,8''-((1,3-phenylenebis(methylene))bis(oxy))(11aS,11a''S)-bis(11-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)-carboxylate)(1-8a)

[0312] To a solution of 1-5b (50 mg, 0.13 mmol) and 1,3-bis(bromomethyl)benzene (17.5 mg, 0.07 mmol) in DMF (2 mL), K2CO3 (36.94 mg, 0.27 mmol) was added at room temperature. The mixture was stirred at 20 °C for 3 hours. The reaction mixture was diluted with  (10 mL) and washed with brine (8 mL x 3). The organic layer was dried over Na2SO4 and concentrated to obtain a residue, which was purified by silica column gel chromatography (eluate: petroleum ether /  = 100 / 0~10 / 90) to obtain 1-8a (50 mg, yield 78.2%) as a white solid.

[0313] MS(ESI)m / z:851.3[M+H] + .

[0314] Step 2: (11aS,11a''S)-8,8''-((1,3-phenylenebis(methylene))bis(oxy))bis(7-methoxy-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one)(1-8)

[0315] To a solution of 1-8a (50 mg, 0.06 mmol) and CH2Cl2 (2 mL), Pd(PPh3)4 (6.79 mg, 0.01 mmol) and pyrrolidine (12 μL, 0.15 mmol) were added at room temperature. The mixture was stirred at 20°C for 30 minutes. The solvent was evaporated, and the residue was purified by silica column gel chromatography (eluate: CH2Cl2 / MeOH = 100 / 0~1 / 99) and preparative HPLC (0.01% FA) to obtain product 1-8 (33.2 mg, yield 78.63%) as a white solid.

[0316] MS(ESI)m / z:647.3[M+H] + .

[0317] 1H NMR(400MHz,CDCl3)δ7.80-7.75(2 H,m),7.54(2 H,s),7.51(1 H,s),7.40(3 H,d,J=4.9),6.85(2 H,d,J 1.4),5.25-5.14(4 H,m),3.96(6 H,s),3.85(2 H,dd,J=7.9,3.2),3.67(2 H,d,J=11.7),3.52-3.45(2 H,m),2.51(2 H,dd,J=13.0,8.1),1.99(2 H,d,J=12.8),0.74(10 H,dd,J=10.6,6.6).

[0318] Examples 1-9 and 1-10 [ka] Step 1: (S)-7-Methoxy-8-((3-(((((S)-7-Methoxy-5-oxo-5,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)methyl)benzyl)oxy)-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one(1- 9) and (S)-7-methoxy-8-((3-(((((S)-7-methoxy-5-oxo-5,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)methyl)benzyl)oxy)-1,10,11,11a-tetrahydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one(1-10)

[0319] To a solution of 1-8 (30 mg, 0.05 mmol) in CH2Cl2 (3 mL), NaBH(OAc)3 (14.76 mg, 0.07 mmol) was added at 0°C. The mixture was stirred at 20°C for 30 minutes. The reaction product was quenched with water (10 mL) and extracted with CH2Cl2 (10 mL x 3). The combined organic layer was dried over Na2SO4 and filtered. The solvent was evaporated, and the residue was purified by preparative HPLC (0.01% FA) to obtain products 1-9 (8 mg, yield 25.25%) as a white solid and 1-10 (12 mg, 38.96%) as a white solid.

[0320] 1-9 MS(ESI)m / z:649.3[M+H] + .

[0321] 1-10 MS(ESI)m / z:651.3[M+H] + .

[0322] 1-10 1 H NMR(400MHz,CDCl3)δ7.57(2 H,s),7.56-7.48(1 H,m),7.37(2 H,d,J=9.8),7.34(3 H,s),6.08(2 H,s),5.13-5.02(4 H,m),3.96( 2 H,t,J=7.1),3.86(6 H,s),3.68(2 H,d,J=12.0),3.54(2 H,d,J=12.0),3.50-3.46(2 H,m),3.38(2 H,dd,J=12.4,8.9),2.06-1.94(2 H,m),1.75(2 H,dd,J=12.6,6.7),0.74-0.51(8 H,m).

[0323] Examples 1-11 [ka] Step 1: Bicyclo[1.1.1]pentane-1,3-diyldimethanol(1-11b)

[0324] LiAlH4 (2.1 g, 52.9 mmol) was added in batches to a solution of 1-11a (3 g, 17.64 mmol) in dry THF (80 ml) at 0°C. The mixture was then warmed to room temperature and stirred overnight. After the reaction was complete, the reaction mixture was quenched with sodium sulfate decahydrate for 1 hour, filtered, and the filtrate was concentrated under reduced pressure to obtain the oil product 1-11b (2.2 g, 97% yield).

[0325] 1 H NMR (400MHz, d6-DMSO) δ4.39(t,J=5.6Hz,2H),3.34(t,J=4.9Hz,4H),1.45(s,6H).

[0326] Step 2: 1,3-Bis(bromomethyl)bicyclo[1.1.1]pentane(1-11c)

[0327] To a solution of triphenylphosphine (2.05 g, 7.81 mmol) in MeCN (30 mL), a solution of liquid bromine (0.4 mL, 7.81 mmol) in 5 mL of MeCN was added dropwise at 0°C, followed by the addition of 1-11b (500 mg, 3.90 mmol). The reaction mixture was heated to 80°C and refluxed overnight. The solvent was removed under vacuum, and the mixture was purified by flash column chromatography to obtain 1-11c (770 mg, 78% yield).

[0328] 1 H NMR (400MHz, CDCl3) δ3.47(s,4H),1.73(s,6H).

[0329] Step 3: Allyl(11S,11As)-8-((3-(bromomethyl)bicyclo[1,1,1]pentan-1-yl)methoxy)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-11d)

[0330] To a solution of 1-7f (30 mg, 0.06 mmol) and 1-11c (154 mg, 0.61 mmol) in 2 mL of DMF, K2CO3 (12 mg, 0.07 mmol) was added at room temperature. The mixture was stirred at room temperature for 3 hours. The mixture was diluted with siRNA and washed with H2O and brine. The organic phase was concentrated and purified by flash column chromatography to obtain 1-11d (40 mg, 99% yield). MS(ESI) m / z: 661.3 [M+H] + .

[0331] Step 4: Allyl(11S,11aS)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-8-((3-((((S)-2-methoxy-14-oxo-6a,7,12,14-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-3-yl)oxy)methyl)bicyclo[1,1,1]pentan-1-yl)methoxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-11e)

[0332] 1-1b (19 mg, 0.06 mmol) and K2CO3 (11 mg, 0.07 mmol) were added to a solution of 1-11d (43 mg, 0.06 mmol) in 1 mL of DMF. The reaction mixture was heated to 40°C for 36 hours. The mixture was diluted with ethyl acetate and washed with water and brine. The organic phase was concentrated and purified by flash column chromatography (CH2Cl2 / MeOH = 94 / 6) to obtain 1-11e (28 mg, 51% yield). MS(ESI) m / z: 889.6 [M+H].

[0333] Step 5: Allyl(11S,11aS)-11-hydroxy-7-methoxy-8-((3-((((S)-2-methoxy-14-oxo-6a,7,12,14-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-3-yl)oxy)methyl)bicyclo[1.1.1]pentan-1-yl)methoxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-11f)

[0334] A buffer of 1 M TBAF (158 μL, 0.16 mmol) and AcOH (11 μL, 0.19 mmol) was added to a solution of 1-11e (28 mg, 0.03 mmol) in 5 mL of dry THF. The mixture was stirred at room temperature for 2 hours. After the reaction was completed, as measured by LC-MS, the mixture was diluted with SiO2O and washed with H2O, 5% citrate, and brine. The organic phase was concentrated and purified by flash column chromatography to obtain 1-11f (24 mg, 98% yield). MS(ESI) m / z: 775.4[M+H] + .

[0335] Step 6: (S)-2-Methoxy-3-((3-(((((S)-7-Methoxy-5-oxo-5,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)methyl)bicyclo[1.1.1]pentan-1-yl)methoxy)-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1,11)

[0336] Pd(PPh3)4 (3.6 mg, 0.003 mmol) was added to a solution of 1-11f (24 mg, 0.03 mmol) in CH2Cl2 (0.5 mL) and pyrrolidine (11 μL, 0.15 mmol). The reaction mixture was stirred at room temperature for 0.5 hours. The reaction product was concentrated and purified by preparative HPLC (0.01% FA in H2O) to obtain 1-11 (10.1 mg, yield 53%) as a white solid. MS(ESI) m / z: 673.3 [M+H] + .

[0337] 1 H NMR(400MHz,CDCl3)δ7.78(d,J=4.1Hz,1H),7.51(d,J=6.8Hz,2H),7.46(d,J=5.0Hz,1H),7.42-7.29(m,4H),6.7 9(d,J=5.2Hz,2H),5.00(d,J=15.5Hz,1H),4.56(d,J=15.4Hz,1H),4.25-4.07(m,4H),3.96-3.94(m,1H),3.94(d, J=2.0Hz,6H),3.85(d,J=5.4Hz,1H),3.67(d,J=11.6Hz,1H),3.49(d,J=11.6Hz,1H),3.27(dd,J=15.2,5.3Hz,1H ),3.16(dd,J=15.4,3.9Hz,1H),2.52(dd,J=13.0,7.9Hz,1H),2.07-1.97(m,1H),1.90(s,6H),0.80-0.68(m,4H).

[0338] Examples 1-12 [ka] Step 1: Allyl(11S,11aS)-8-((3-(bromomethyl)bicyclo[1,1,1]pentan-1-yl)methoxy)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-12a)

[0339] To a solution of 1-7f (35 mg, 0.07 mmol) and 1-11c (180 mg, 0.72 mmol) in 2 mL of DMF, K2CO3 (12 mg, 0.09 mmol) was added at room temperature. The mixture was stirred at room temperature for 5 hours. The mixture was diluted with siRNA and washed with H2O and brine. The organic phase was concentrated and purified by flash column chromatography to obtain 1-12a (41 mg, 86% yield). MS(ESI) m / z: 661.3 [M+H] + .

[0340] Step 2: Allyl(S)-3-((3-((((11S,11aS)-10-((allyloxy)carbonyl)-11-((tert-butyldimethylsilyl)oxy)-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)methyl)bicyclo[1.1.1]pentan-1-yl)methoxy)-2-methoxy-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(1-12b)

[0341] 1-2c (23 mg, 0.06 mmol) and K2CO3 (11 mg, 0.08 mmol) were added to a solution of 1-12a (41 mg, 0.06 mmol) in 1 mL of DMF. The mixture was stirred overnight at room temperature. The product (petroleum ether / siRNA = 1:2) was detected by LC-MS. The mixture was diluted with siRNA and washed with water and brine. The organic phase was concentrated and purified by flash column chromatography (petroleum ether / siRNA = 27 / 73) to obtain 1-12b (48 mg, 84% yield). MS(ESI) m / z: 4974.6[M+H] + .

[0342] Step 3: Allyl(S)-3-((3-((((11S,11aS)-10-((allyloxy)carbonyl)-11-hydroxy-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)methyl)bicyclo[1.1.1]pentan-1-yl)methoxy)-2-methoxy-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(1-12c)

[0343] A mixed solution of 1 M TBAF (248 μL, 0.25 mmol) and AcOH (18 μL, 0.30 mmol) was added to a solution of 1-12b (48 mg, 0.05 mmol) in 5 mL of dry THF. The mixture was stirred at room temperature for 2 hours. After the reaction was complete by LC-MS, the mixture was diluted with SiO2O and washed with H2O, 5% citric acid, and brine. The organic phase was concentrated and purified by flash column chromatography to obtain 1-12c (30 mg, 78% yield). MS(ESI) m / z: 861.5[M+H] + .

[0344] Step 4: (S)-2-Methoxy-3-((3-(((((S)-7-Methoxy-5-oxo-5,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)methyl)bicyclo[1.1.1]pentan-1-yl)methoxy)-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(5H)-one(1-12)

[0345] Pd(PPh3)4 (4.0 mg, 0.003 mmol) was added to a solution of 1-12c (30 mg, 0.03 mmol) in CH2Cl2 (0.5 mL) and pyrrolidine (13 μL, 0.17 mmol). The reaction mixture was stirred at room temperature for 0.5 hours. The reaction product was concentrated and purified by preparative HPLC (0.01% FA in H2O) to obtain 1-12c (18 mg, 77% yield) as a white solid. MS(ESI) m / z: 675.4 [M+H] + .

[0346] 1 H NMR(400MHz,CDCl3)δ7.78(d,J=4.4Hz,1H),7.71-7.63(m,2H),7.57-7.53(m,1H),7.51-7.43(m,3H),7.34-7.32(m, 1H),7.21-7.15(m,1H),6.80(s,1H),4.87(d,J=15.7Hz,1H),4.71(d,J=15.7Hz,1H),4.21-4.02(m,6H),3.94(s,3H) ,3.84-3.82(s,3H),3.67(d,J=11.7Hz,1H),3.53-3.39(m,2H),3.21(t,J=10.9Hz,1H),3.10(dd,J=15.2,5.7Hz,1H) ,2.80(dd,J=15.2,5.0Hz,1H),2.52(dd,J=13.0,8.1Hz,1H),2.00(dd,J=13.1,2.6Hz,1H),1.89(s,6H),0.74(m,4H).

[0347] Examples 1-13 [ka] Step 1: Allyl bis(2-hydroxyethyl)carbamate (1-13b)

[0348] To a solution of 1-13a (423 mg, 3 mmol) in THF (3.1 mL) and water (5.7 mL), AllocCl (361.45 mg, 3 mmol) and K2CO3 (1036 mg, 7.5 mmol) were added at 0°C. The mixture was stirred at 20°C for 16 hours. TLC (petroleum ether: Â=1:1, v / v) indicated that the reaction was complete. The reaction mixture was poured into water (10 mL) and extracted with  (20 mL x 3). The combined organic layer was dried over Na2SO4 and filtered. The solvent was evaporated, and the crude product 1-13b (567 mg) was used in the next step without further processing or purification.

[0349] Step 2: (((allyloxy)carbonyl)azandiyl)bis(ethane-2,1-diyl)bis(4-methylbenzenesulfonate)(1-13c)

[0350] To a solution of 1-13b (567 mg, 3 mmol) in CH2Cl2 (7 mL), TsCl (1.7 g, 9 mmol) and triethylamine (1.67 mL, 12 mmol) were added at 0°C. The mixture was stirred at 20°C for 16 hours. TLC (petroleum ether: siRNA = 3:1, v / v) indicated that the reaction was complete. The reaction product was poured into water (20 mL) and extracted with siRNA (30 mL x 3). The combined organic layer was dried over Na2SO4 and filtered. The solvent was evaporated, and the residue was purified by silica column gel chromatography (eluate: petroleum ether / siRNA = 100 / 0~30 / 70) to obtain 1-13c (1.2 g, yield 73.08%) as a colorless oil.

[0351] Step 3: diallyl 8,8''-((((((allyloxy)carbonyl)azandiyl)bis(ethane-2,1-diyl))bis(oxy))(11aS,11a''S)-bis(11-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)-carboxylate)(1-13d)

[0352] A solution of 1-13c (50 mg, 0.10 mmol) and 1-5b (75.25 mg, 0.20 mmol) in DMSO (2 mL) was mixed with K2CO3 (41.70 mg, 0.30 mmol) at room temperature. The mixture was stirred at 50°C for 16 hours. LC-MS indicated that the reaction was complete. The reaction mixture was diluted with  (10 mL) and washed with brine (8 mL x 3). The organic layer was dried over Na2SO4 and concentrated to obtain a residue, which was purified by silica column gel chromatography (eluate: petroleum ether /  = 100 / 0~10 / 90) to obtain 1-13d (38 mg, yield 49.64%) as a colorless oil.

[0353] MS(ESI)m / z:902.3[M+H] + .

[0354] Step 4: (11aS,11a''S)-8,8''-((Azandiylbis(ethane-2,1-diyl))bis(oxy))bis(7-methoxy-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one)(1-13)

[0355] To a solution of 1-13d (45 mg, 0.05 mmol) and CH2Cl2 (3 mL), Pd(PPh3)4 (5.77 mg, 0.005 mmol) and pyrrolidine (12 μL, 0.12 mmol) were added at room temperature. The mixture was stirred at 20°C for 30 minutes. LC-MS indicated that the reaction was complete. The solvent was evaporated, and the residue was purified by preparative HPLC (0.01% FA) to obtain product 1-13 (12 mg, yield 35.27%) as a white solid.

[0356] MS(ESI)m / z:614.3[M+H] + .

[0357] 1H NMR(400MHz,CDCl3)δ7.79(2 H,d,J=4.4),7.51(2 H,s),6.84(2 H,d,J=2.2),4.30-4.13(4 H,m),3.93(6 H,d,J=0.9),3.87-3.81(2 H,m),3.67(2 H,d,J=11.7),3.49(2 H,d,J=11.7),3.26-3.17(4 H,m),2.52(2 H,dd,J=13.0,8.1),2.00(2 H,dd,J=13.0,2.6),0.73(8 H,ddd,J=11.3,8.3,4.5).

[0358] Examples 1-14 [ka] Step 1: (S)-3-(3-bromopropoxy)-2-methoxy-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1-14a)

[0359] To a solution of 1-1b (83 mg, 0.27 mmol) and 1,3-dibromopropane (0.42 mL, 4.04 mmol) in 2 mL of DMF, K2CO3 (45 mg, 0.32 mmol) was added at room temperature. The mixture was stirred at room temperature for 2 hours. The mixture was diluted with siRNA and washed with H2O and brine. The organic phase was dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (CH2Cl2 / MeOH = 96 / 4) to obtain 1-14a (98 mg, yield 85%).

[0360] MS(ESI)m / z:429.1[M+H] + .

[0361] Step 2: Allyl(11S,11aS)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-8-(3-(((S)-2-methoxy-14-oxo-6a,7,12,14-tetrahydrobenzo[5,6][1,4]diazepine[1,2-b]isoquinoline-3-yl)oxy)propoxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-14b)

[0362] 1-7f (25 mg, 0.06 mmol) and K2CO3 (10.5 mg, 0.08 mmol) were added to a solution of 1-14a (43 mg, 0.09 mmol) in 1 mL of DMF. The mixture was stirred at 40°C for 36 hours. The mixture was diluted with siRNA and washed with water and brine. The organic phase was dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (CH2Cl2 / MeOH = 96 / 4) to obtain 1-14b (40 mg, yield 82%).

[0363] MS(ESI)m / z:837.5[M+H] + .

[0364] Step 3: Allyl(11S,11aS)-11-hydroxy-7-methoxy-8-(3-(((S)-2-methoxy-14-oxo-6a,7,12,14-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-3-yl)oxy)propoxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-14c)

[0365] 1M TBAF (240 μL, 0.24 mmol) was added to a solution of 1-14b (40 mg, 0.05 mmol) in 1.5 mL of dry THF and AcOH (16 μL, 0.29 mmol). The mixture was stirred at room temperature for 2 hours. The mixture was diluted with ethyl acetate and washed with saturated NaHCO3 and brine. The organic phase was dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (CH2Cl2 / MeOH = 96 / 4) to obtain 1-14c (30 mg, 77% yield).

[0366] MS(ESI)m / z:723.4[M+H] + .

[0367] Step 4: (S)-2-Methoxy-3-(3-((((S)-7-Methoxy-5-oxo-5,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)propoxy)-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1,14)

[0368] Pd(PPh3)4 (3.84 mg, 0.003 mmol) was added to a solution of 1-14c (24 mg, 0.033 mmol) in CH2Cl2 (1 mL) and pyrrolidine (6.82 μL, 0.083 mmol). The reaction mixture was stirred at room temperature for 20 minutes. The reaction product was concentrated and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*150 mm; Mobile phase: A-Water (without formic acid): B-Acetonitrile; Flow rate: 20 mL / min). The fraction was lyophilized to obtain 1-14 (6.5 mg, yield 32%) as a white solid.

[0369] MS(ESI)m / z:621.4[M+H] + .

[0370] Examples 1-15 [ka] Step 1: Allyl(S)-7-methoxy-5-oxo-8-((triisopropylsilyl)oxy)-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-15a)

[0371] DIAD (1.14 mL, 5.75 mmol) was added to a solution of 1-7c (1.53 g, 2.87 mmol) and PPh3 (2.26 g, 8.62 mmol) in THF (30 mL). The reaction mixture was stirred under N2 at 40°C for 1 hour. The solvent was removed under vacuum, and the residue was purified by silica gel column chromatography (petroleum ether / siRNA = 30 / 70) to obtain 1-15a (940 mg, yield 64%).

[0372] MS(ESI)m / z:515.4[M+H] + .

[0373] Step 2: Allyl(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)-carboxylate(1-15b)

[0374] Lithium acetate (121 mg, 1.83 mmol) was added to a solution of 1-15a (940 mg, 1.83 mmol) in wet DMF (15 mL, DMF / water = 49 / 1). The reaction mixture was stirred at 25°C for 2 hours. The mixture was diluted with ethyl acetate and washed twice with H2O and brine. The organic phase was dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (CH2Cl2 / MeOH = 96 / 4) to obtain 1-15b (605 mg, yield 92%).

[0375] MS(ESI)m / z:359.2[M+H] + .

[0376] Step 3: Allyl(S)-7-methoxy-8-(3-(((S)-2-methoxy-14-oxo-6a,7,12,14-tetrahydrobenzo[5,6][1,4]diazepine[1,2-b]isoquinoline-3-yl)oxy)propoxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-15c)

[0377] 1-15b (26 mg, 0.06 mmol) and K2CO3 (10.03 mg, 0.07 mmol) were added to a solution of 1-14a (20 mg, 0.06 mmol) in 1 mL of DMF. The mixture was stirred overnight at room temperature. The mixture was diluted with siRNA and washed with water and brine. The organic phase was dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (CH2Cl2 / MeOH = 96 / 4) to obtain 1-15c (33 mg, yield 84%).

[0378] MS(ESI)m / z:707.4[M+H] + .

[0379] Step 4: (S)-2-Methoxy-3-(3-((((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)propoxy)-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1,15)

[0380] Pd(PPh3)4 (5.4 mg, 0.005 mmol) was added to a solution of 1-15c (33 mg, 0.047 mmol) in CH2Cl2 (1 mL) and pyrrolidine (9.6 μL, 0.117 mmol). The reaction mixture was stirred under N2 at room temperature for 0.5 hours. The reaction product was concentrated and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*150 mm; Mobile phase: A-Water (without formic acid): B-Acetonitrile; Flow rate: 20 mL / min). The fraction was lyophilized to obtain 1-15 (5.1 mg, yield 17%) as a white solid.

[0381] MS(ESI)m / z:623.4[M+H] + .

[0382] Examples 1-16 [ka] Step 1: (S)-(4-(benzyloxy)-5-methoxy-2-nitrophenyl)(2-(hydroxymethyl)piperidine-1-yl)methanone(1-16b)

[0383] Oxalyl chloride (0.43 mL, 4.94 mmol) was added dropwise to a stirred solution containing 1-6 g (600 mg, 1.97 mmol) in CH2Cl2 (5.1 mL), THF (0.51 mL), and DMF (2.4 μL, 0.031 mmol) at 0°C under N2 conditions. The reaction mixture was warmed to room temperature and stirred for 1 hour. The reaction mixture was concentrated to obtain a yellowish-white solid, which was used in the next step without purification.

[0384] The obtained solid and 1-16a (250.6 mg, 2.18 mmol) were dissolved in CH2Cl2 (5.6 mL), and the reaction mixture was then cooled to 0°C. Et3N (0.4 mL, 2.96 mmol) was added dropwise under N2. The reaction mixture was then warmed to room temperature and stirred for 2 hours. The reaction mixture was concentrated to obtain the crude product, which was purified by silica gel chromatography (eluate: siRNA / hexane = 0%~80%) to obtain 1-16b (720 mg, yield 91%) as a yellow solid.

[0385] MS(ESI)m / z:401.16[M+H] + .

[0386] Step 2: (S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)piperidine-2-carbaldehyde(1-16c)

[0387] DMP (805 mg, 1.89 mmol) was slowly and gradually added at 0°C to a solution of 1-16b (700 mg, 1.75 mmol) in CH2Cl2 (7 mL). The reaction mixture was then warmed to room temperature and stirred for 3 hours. The reaction mixture was filtered, and the filtrate was washed with saturated sodium thiosulfate aqueous solution (10 mL). Subsequently, saturated NaHCO3 aqueous solution (10 mL) and H2O (10 mL) were slowly added. The mixture was extracted with CH2Cl2 (10 mL x 3), the organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated to obtain the title compound 1-16c (730 mg, crude) as a yellow solid, which was used in the next step without purification.

[0388] MS(ESI)m / z:399.15[M+H] + .

[0389] Step 3: (S)-3-(benzyloxy)-2-methoxy-7,8,9,10-tetrahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-12(6aH)-one(1-16d)

[0390] Compound 1-16c (730 mg, 1.83 mmol) was dissolved in a mixed solvent of THF (0.9 mL), methanol (4.5 mL), and water (0.9 mL). Then, NH4Cl (980.06 mg, 18.32 mmol) and iron powder (511.6 mg, 9.16 mmol) were added. The reaction mixture was then heated to 50°C under N2 and stirred for 16 hours. The reaction mixture was filtered through Celite. The filtrate was diluted with water (5 mL) and extracted with Â(10 mL x 3). The organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated to obtain the crude product, which was purified by silica gel chromatography (eluate: Â(0%) / hexane = 0%~60%) to obtain 1-16d (500 mg, yield 77.8%) as a yellow solid.

[0391] MS(ESI)m / z:351.2[M+H] + .

[0392] Step 4: (S)-3-hydroxy-2-methoxy-7,8,9,10-tetrahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-12(6aH)-one(1-16e)

[0393] MeSO3H (0.37 mL, 5.7 mmol) was added dropwise to a solution of 1-16d (200 mg, 0.57 mmol) in CH2Cl2 (2.5 mL) at 0°C. The reaction mixture was stirred at 0°C for 10 minutes, then warmed to room temperature and stirred for 2 hours. The mixture was quenched with saturated NaHCO3 (10 mL), then extracted with CH2Cl2 (5 mL x 3), the organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated to obtain the crude product. This was purified by silica column gel chromatography (eluate: hexane / Â=100 / 0~20 / 80) to obtain 1-16e (120 mg, yield 80.7%) as a white solid.

[0394] MS(ESI)m / z:261.2[M+H] + .

[0395] Step 5: (S)-3-((5-iodopentyl)oxy)-2-methoxy-7,8,9,10-tetrahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-12(6aH)-one(1-16f)

[0396] 1,5-Diiodopentane (0.34 mL, 2.31 mmol) was added to a solution of 1-16e (120 mg, 0.46 mmol) in dry DMF (2 mL). The solution was then cooled to 0°C, and K2CO3 (127 mg, 0.92 mmol) was added all at once. The reaction mixture was stirred at room temperature for 6 hours. siRNA (20 mL) and H2O (10 mL) were added, the organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated to obtain the crude product. This was purified by silica column gel chromatography (eluate: hexane / siRNA = 100 / 0~50 / 50) to obtain 1-16f (130 mg, yield 61.7%) as a yellow solid.

[0397] MS(ESI)m / z:457.1[M+H] + .

[0398] Step 6: Allyl(11S,11aS)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-8-((5-(((S)-2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-3-yl)oxy)pentyl)oxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate (1-16g)

[0399] To a solution of 1-7f (50 mg, 0.102 mmol) and 1-16f (51.3 mg, 0.11 mmol) in DMF (0.5 mL), K2CO3 (16.9 mg, 0.12 mmol) was added. The mixture was stirred at room temperature for 3 hours. Â (15 mL) and H2O (10 mL) were added, the organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated to obtain the crude product. This was purified by silica column gel chromatography (eluate: hexane / Â = 100 / 0~50 / 50) to obtain 1-16 g (60 mg, yield 71.7%) as a white solid.

[0400] MS(ESI)m / z:817.5[M+H] + .

[0401] Step 7: Allyl(11S,11aS)-11-hydroxy-7-methoxy-8-((5-(((S)-2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-3-yl)oxy)pentyl)oxy)-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-16h)

[0402] To a solution of 1-16 g (60 mg, 0.07 mmol) in dry THF (1 mL), AcOH (24 μL) was added, followed by dropwise addition of TBAF (350 μL, 0.35 mmol, 1 M in THF). The mixture was stirred at room temperature for 6 hours. The mixture was quenched with saturated NaHCO3, extracted with  (5 mL x 3), the organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated to obtain the crude product. This was purified by silica column gel chromatography (eluate: CH2Cl2 / MeOH = 100 / 0~5 / 95) to obtain 1-16 h (40 mg, yield 77.5%) as a white solid.

[0403] MS(ESI)m / z:703.4[M+H] + .

[0404] Step 8: (S)-7-Methoxy-8-((5-((((S)-2-Methoxy-12-oxo-6a,7,8,9,10,12-Hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-3-yl)oxy)pentyl)oxy)-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one(1-16)

[0405] To a solution of 1-16h (40 mg, 0.056 mmol) in CH2Cl2 (1 mL), Pd(PPh3)4 (2.0 mg, catalytic amount) and pyrrolidine (11 μL, 0.14 mmol) were added. The reaction mixture was stirred for 15 minutes under N2 at room temperature. The reaction product was neutralized with AcOH and concentrated to obtain a residue, which was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*150 mm; Mobile phase: A-Water (0.01% formic acid): B-Acetonitrile; Flow rate: 20 mL / min) to obtain 1-16 (9.6 mg, yield 28.1%) as a white solid.

[0406] MS(ESI)m / z:601.1[M+H] + .

[0407] 1 H NMR(400MHz,CDCl3)δ7.92(s,1H),7.81(s,1H),7.52(s,1H),7.44(s,1H),6.79 (d,J=14.2Hz,2H),4.24(s,1H),4.10(d,J=22.7Hz,4H),3.94(d,J=3.5Hz,6H), 3.87(s,1H),3.79(s,1H),3.69(d,J=11.3Hz,1H),3.51(d,J=12.2Hz,1H),3.25 (s,1H),2.62-2.47(m,1H),2.19-1.77(m,11H),1.68(s,4H),0.83-0.64(m,4H).

[0408] Examples 1-17 [ka] Step 1: diallyl 8,8''-((pyridine-2,6-diylbis(methylene))bis(oxy))(11S,11aS,11''S,11a''S)-bis(11-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)-carboxylate)(1-17b)

[0409] Potassium carbonate (84 mg, 0.77 mmol) was added to a solution of 1-5b (84 mg, 0.23 mmol) and 1-17a (30 mg, 0.11 mmol) in 3 mL of DMF, and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic phase was washed with water and brine and dried over anhydrous sodium sulfate. The organic phase was then concentrated under vacuum to obtain the crude product, which was used directly in the next step without further purification.

[0410] MS(ESI)m / z:852.9[M+H] + .

[0411] Step 2: (11aS,11a''S)-8,8''-((pyridine-2,6-diylbis(methylene))bis(oxy))bis(7-methoxy-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one)(1-17)

[0412] To a solution of 1-17b (84 mg, 0.23 mmol) in THF / CH2Cl2 (1 mL / 1 mL), Pd(PPh3)4 (12 mg, 0.01 mmol) and 1,3-dimedone (32 mg, 0.23 mmol) were added, and the mixture was stirred at room temperature for 1 hour. The solution was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*150 mm; Mobile phase: A-Water (0.01% formic acid): B-Acetonitrile; Flow rate: 20 mL / min) to obtain 1-17 (36 mg, yield 50%) as a white solid.

[0413] 1 H NMR(400MHz,CDCl3)δ7.81(m,3H),7.61(s,2H),7.53(d,J=7.7Hz,2H),6.92(s,2H),5.85-5.03 (m,4H),4.27-3.96(m,6H),3.96(m,2H),3.73(d,J=11.7Hz,2H),3.55(d,J=11.7Hz,2H),2.57(d J=12.9,2H),2.05(d,J=12.8Hz,2H),0.93-0.71(m,8H).

[0414] MS(ESI)m / z:648.7[M+H] + .

[0415] Examples 1-18 [ka] Step 1: 5-((tert-butyldimethylsilyl)oxy)isophthalic acid (1-18b)

[0416] To a solution of 1-18a (0.5 g, 2.74 mmol) in 10 mL of DMF, TBS-Cl (2 g, 13.7 mmol) and imidazole (1.12 g, 16.4 mmol) were added, and the mixture was stirred at 50°C for 4 hours. The mixture was acidified to pH=3 with 1N HCl, diluted with water (10 mL), and extracted with ethyl acetate (10 mL x 2). The combined organic phase was washed with water and brine and dried over anhydrous sodium sulfate. The organic phase was then concentrated under vacuum to obtain a residue, which was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 50) to obtain 1-18b (170 mg, yield 21%) as a white solid.

[0417] MS(ESI)m / z:297.4[M+H] + .

[0418] Step 2: (5-((tert-butyldimethylsilyl)oxy)-1,3-phenylene)dimethanol(1-18c)

[0419] To a solution of 1-18b (50 mg, 0.17 mmol) in 2 mL of THF, LiAlH4 (1 M, 0.34 mL, 0.34 mmol) was added, and the mixture was stirred at 40°C for 2 hours. The mixture was quenched with H2O (36 μL), 10% NaOH aqueous solution (40 μL), and H2O (200 μL), and the mixture was stirred at room temperature for 1 hour. The solution was filtered, and the filtrate was concentrated under vacuum to obtain the residue. The residue was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*150 mm; Mobile phase: A-Water (0.1% formic acid): B-Acetonitrile; Flow rate: 20 mL / min) to obtain 1-18c (27 mg, yield 27.1%) as a white solid.

[0420] MS(ESI)m / z:269.4[M+H] + .

[0421] Step 3: (3,5-bis(bromomethyl)phenoxy)(tert-butyl)dimethylsilane (1-18d)

[0422] A solution of 1-18c (21 mg, 0.08 mmol) in 2 mL of CH3CN was cooled to 0°C, followed by the addition of PPh3 (62 mg, 0.24 mmol) and CBr4 (78 mg, 0.24 mmol). The mixture was stirred at room temperature for 1 hour. The solution was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic phase was washed with water and brine and dried over anhydrous sodium sulfate. The organic phase was then concentrated under vacuum to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 90 / 10) to obtain 1-18d (30 mg, yield 96%) as a colorless oil.

[0423] MS(ESI)m / z:395.2[M+H] + .

[0424] Step 4: Allyl(11S,11aS)-8-((3-(((11aS)-10-((allyloxy)carbonyl)-11-((tert-butyldimethylsilyl)oxy)-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)methyl)-5-hydroxybenzyl)oxy)-11-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(1-18e)

[0425] Potassium carbonate (23 mg, 0.16 mmol) was added to a solution of 1-18d (30 mg, 0.08 mmol) and 1-7f (79 mg, 0.16 mmol) in 3 mL of DMF, and the mixture was stirred at room temperature for 4 hours. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic phase was washed with water and brine and dried over anhydrous sodium sulfate. The solution was concentrated to obtain a residue, which was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 90 / 10) to obtain 1-18e (58 mg, yield 63%) as a white solid.

[0426] MS(ESI)m / z:1096.5[M+H] + .

[0427] Step 5: diallyl 8,8''-(((5-hydroxy-1,3-phenylene)bis(methylene))bis(oxy))(11S,11aS,11''S,11a''S)-bis(11-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)-carboxylate)(1-18f)

[0428] To a solution of 1-18e (58 mg, 0.05 mmol) in 3 mL of THF, TBAF (159 μL, 0.16 mmol) and AcOH (15 μL, 0.27 mmol) were added dropwise, and the mixture was stirred at room temperature for 1 hour. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic phase was washed with water and brine and dried over anhydrous sodium sulfate. The solution was concentrated to obtain a residue, which was purified by silica gel column chromatography (CH2Cl2 / MeOH = 90 / 10) to obtain 1-18f (42 mg, yield 91.3%) as a white solid.

[0429] MS(ESI)m / z:867.9[M+H] + .

[0430] Step 6: (11aS,11a''S)-8,8''-(((5-hydroxy-1,3-phenylene)bis(methylene))bis(oxy))bis(7-methoxy-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one)(1-18)

[0431] To a solution of 1-18f (42 mg, 0.05 mmol) in CH2Cl2 (2 mL), Pd(PPh3)4 (3 mg, 0.003 mmol) and pyrrolidine (21 μL, 0.25 mmol) were added, and the mixture was stirred at room temperature for 1 hour. The solution was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*150 mm; Mobile phase: A-Water (0.01% formic acid): B-Acetonitrile; Flow rate: 20 mL / min) to obtain 1-18 (18 mg, yield 55%) as a white solid.

[0432] 1 H NMR(400MHz,CDCl3)δ7.75(s,2H),7.48(s,2H),6.90(m,5H),5.08(m,4H),4.01-3.79(m,8H),3.6 6(d,J=11.8Hz,2H),3.47(d,J=11.7Hz,2H),2.69-2.37(m,2H),2.02-1.89(m,2H),0.78(dm,8H).

[0433] MS(ESI)m / z:663.7[M+H] + .

[0434] Examples 1-19 [ka] Step 1: diallyl 8,8''-(((E)-pento-2-ene-1,5-diyl)bis(oxy))(11S,11aS,11''S,11a''S)-bis(11-((tert-butyldimethylsilyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate)(1-19b)

[0435] PPh3 (96 mg, 0.37 mmol) and DIAD (74 μL, 0.37 mmol) were dissolved in THF (3 mL) and stirred at room temperature for 2 hours. Subsequently, 1-7f (107 mg, 0.22 mmol) and 1-19a (8 mg, 0.07 mmol) were added, and the mixture was stirred at room temperature for a further 1 hour. The solution was concentrated, and the crude product was purified by silica gel column chromatography (CH2Cl2 / Â=50 / 50) to obtain 1-19b (33 mg, yield 15%) as a white solid.

[0436] MS(ESI)m / z:1044.4[M+H] + .

[0437] Step 2: Allyl(11S,11aS)-8-(((E)-5-(((11S,11aS)-10-((allyloxy)carbonyl)-11-((tert-butyldimethylsilyl)oxy)-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)pento-2-en-1-yl)oxy)-11-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)-carboxylate(1-19c)

[0438] To a solution of 1-19b (33 mg, 0.03 mmol) in 3 mL of THF, TBAF (106 μL, 0.11 mmol) and AcOH (10 μL, 0.15 mmol) were added dropwise, and the mixture was stirred at room temperature for 1 hour. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic phase was washed with water and brine and dried over anhydrous sodium sulfate. The solution was concentrated to obtain a residue, which was purified by silica gel column chromatography (CH2Cl2 / MeOH = 90 / 10) to obtain 1-19c (21 mg, yield 81%) as a white solid.

[0439] MS(ESI)m / z:815.9[M+H] + .

[0440] Step 3: (11aS,11a''S)-8,8''-(((E)-penta-2-ene-1,5-diyl)bis(oxy))bis(7-methoxy-1,11a-dihydro-3H,5H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-5-one)(1-19)

[0441] To a solution of 1-19c (21 mg, 0.025 mmol) in CH2Cl2 (2 mL), Pd(PPh3)4 (2 mg, 0.0013 mmol) and pyrrolidine (11 μL, 0.13 mmol) were added, and the mixture was stirred at room temperature for 1 hour. The solution was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*150 mm; Mobile phase: A-Water (0.01% formic acid): B-Acetonitrile; Flow rate: 20 mL / min) to obtain 1-19 (4 mg, yield 26%) as a white solid.

[0442] MS(ESI)m / z:611.7[M+H] + .

[0443] Examples 1-20 [ka] Step 1: (S)-6-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-6-azaspiro[2.5]octane-5-carbaldehyde(1-20a)

[0444] To a solution of 1-6h (80 mg, 0.16 mmol) in CH2Cl2 (3 mL), DMP (105 mg, 0.24 mmol) was added at 0°C, then the mixture was warmed to room temperature and stirred for 1 hour. Saturated Na2S2O3 (5 mL) and saturated NaHCO3 (5 mL) were added to the solution, and then extracted with CH2Cl2 (5 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to obtain the residue, which was purified by flash column chromatography (eluent: hexane / siRNA = 100 / 0~50 / 50) to obtain 1-20a (70 mg, yield 90%) as a white solid.

[0445] MS(ESI)m / z:425.1[M+H] + .

[0446] Step 2: (S)-3-(benzyloxy)-2-methoxy-6a,7,9,10-tetrahydro-12H-spiro[benzo[e]pyrido[1,2-a][1,4]diazepine-8,1'-cyclopropane]-12-one(1-20b)

[0447] To a solution of 1-20a (70 mg, 0.16 mmol) in MeOH (3 mL) and H2O (1 mL), NH4Cl (134 mg, 2.46 mmol), followed by iron powder (46.5 mg, 0.81 mmol), was added. The reaction mixture was refluxed under N2 for 3 hours. The reaction mixture was filtered through Celite. The filtrate was diluted with water (5 mL) and extracted with RINKAN (5 mL x 3). The organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated to obtain 1-20b (54 mg, crude), which was used directly in the next step without purification.

[0448] MS(ESI)m / z:377.2[M+H] + .

[0449] Step 3: (S)-3-hydroxy-2-methoxy-6a,7,9,10-tetrahydro-12H-spiro[benzo[e]pyrido[1,2-a][1,4]diazepine-8,1'-cyclopropane]-12-one(1-20c)

[0450] To a solution of 1-20b (50 mg, 0.08 mmol) in CH2Cl2 (2.5 mL), MsOH (55 μL, 0.8 mmol) was added at 0°C, and the mixture was stirred at 0°C for 5 hours. The mixture was quenched with saturated NaHCO3 and extracted with CH2Cl2 (5 mL x 3). The organic layer was washed with brine (5 mL), dried over Na2SO4, and concentrated to obtain the crude product. This was purified by silica column gel chromatography (eluate: hexane / Â=100 / 0~25 / 75) to obtain 1-20c (25 mg, yield 65.7%) as a yellow solid.

[0451] MS(ESI)m / z:287.1[M+H] + .

[0452] Step 4: (S)-2-methoxy-3-((5-((((S)-2-methoxy-12-oxo-6a,9,10,12-tetrahydro-7H-spiro[benzo[e]pyrido[1,2-a][1,4]diazepine-8,1'-cyclopropane]-3-yl)oxy)pentyl)oxy)-7,12-dihydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-14(6aH)-one(1-20)

[0453] To a solution of 1-20c (25 mg, 0.087 mmol) and 1-1c (48.4 mg, 0.096 mmol) in DMF (0.5 mL), K2CO3 (14.5 mg, 0.1 mmol) was added. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was quenched with H2O (15 mL) and extracted with  (10 mL x 3). The organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated to obtain the crude product, which was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19 x 150 mm; Mobile phase: A-Water (0.01% formic acid): B-Acetonitrile; Flow rate: 20 mL / min) to obtain 1-20 (8.9 mg, yield 15.6%) as a white solid.

[0454] MS(ESI)m / z:663.7[M+H] + .

[0455] Examples 1-21 [ka] Step 1: (S)-(2-(((tert-butyldimethylsilyl)oxy)methyl)-4-methylenepyrrolidine-1-yl)(4-hydroxy-5-methoxy-2-nitrophenyl)methanone(1-21b)

[0456] To a mixture of MePh3PBr (24.6 g, 68.86 mmol) in THF (100 mL), t-BuOK (6.95 g, 61.98 mmol) was added under N2 conditions at 0°C. The mixture was stirred at 0°C for 2 hours, then a solution of 1-21a (4 g, 6.89 mmol) in THF (30 mL) was added dropwise to the mixture, and the mixture was stirred at 0°C for 16 hours. The mixture was neutralized with citric acid and extracted with siRNA (100 mL x 3). The organic phase was dried over Na2SO4, filtered, and concentrated to obtain the crude product, which was purified by silica column gel chromatography (eluent: hexane / siRNA = 100 / 0~25 / 75) to obtain 1-21b (1.1 g, yield 37.8%) as a yellow solid.

[0457] MS(ESI)m / z:423.2[M+H] + .

[0458] Step 2: Allylbis(2-(4-((S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-methylenepyrrolidine-1-carbonyl)-2-methoxy-5-nitrophenoxy)ethyl)carbamate(1-21c)

[0459] To a solution of 1-13c (100 mg, 0.2 mmol) and 1-21b (186.83 mg, 0.44 mmol) in DMSO (3 mL), K2CO3 (55.5 mg, 0.4 mmol) was added. The mixture was stirred at 50°C for 16 hours. The reaction product was quenched with H2O and extracted with  (30 mL). The organic phase was dried over Na2SO4, filtered, and concentrated to obtain the crude product, which was purified by silica column gel chromatography (eluate: hexane /  = 100 / 0~0 / 100) to obtain 1-21c (108 mg, yield 53.83%) as a yellow solid.

[0460] MS(ESI)m / z:998.5[M+H] + .

[0461] Step 3: Allylbis(2-(5-amino-4-((S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-methylenepyrrolidine-1-carbonyl)-2-methoxyphenoxy)ethyl)carbamate(1-21d)

[0462] A mixture of Zn powder (28.24 mg, 4.08 mmol) in EtOH (3 mL), AcOH (0.2 mL), and H2O (0.2 mL) was stirred at room temperature for 10 minutes. Subsequently, a solution of 1-21c (108 mg, 0.11 mmol) in EtOH (2 mL) was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was filtered through Celite, and the filtrate was concentrated to obtain the crude product. This was purified by silica gel chromatography (eluate: CH2Cl2 / MeOH = 20 / 1) to obtain 1-21d (73 mg, yield 72.2%) as yellow oil.

[0463] MS(ESI)m / z:938.6[M+H] + .

[0464] Step 4: Allylbis(2-(5-(((allyloxy)carbonyl)amino)-4-((S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-methylenepyrrolidine-1-carbonyl)-2-methoxyphenoxy)ethyl)carbamate(1-21e)

[0465] To a solution of 1-21d (73 mg, 0.077 mmol) in dry CH2Cl2 (2 mL), Alloc-Cl (53 μL, 0.49 mmol) and pyridine (24.73 μL, 0.311 mmol) were added under an N2 atmosphere at -10°C. The mixture was stirred at -10°C for 1 hour. The solution was added to water (10 mL) and extracted with CH2Cl2 (5 mL x 3). The organic phase was dried over Na2SO4, filtered, and concentrated to obtain the crude product, which was purified by flash column chromatography (eluent: petroleum ether / siRNA = 1 / 3) to obtain 1-21e (75 mg, yield 87.13%) as a grayish-white solid.

[0466] MS(ESI)m / z:1106.6[M+H]+ .

[0467] Step 5: Allylbis(2-(5-(((allyloxy)carbonyl)amino)-4-((S)-2-(hydroxymethyl)-4-methylenepyrrolidine-1-carbonyl)-2-methoxyphenoxy)ethyl)carbamate(1-21f)

[0468] To a solution of 1-21e (75 mg, 0.067 mmol) in THF (2 mL) and water (0.1 mL), p-toluenesulfonic acid hydrate (25.79 mg, 0.13 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated and purified by flash column chromatography (eluate: CH2Cl2 / MeOH = 20 / 1) to obtain 1-21f (52 mg, yield 87.38%) as a yellow solid.

[0469] MS(ESI)m / z:878.4[M+H] + .

[0470] Step 6: diallyl 8,8'-((((allyloxy)carbonyl)azandiyl)bis(ethane-2,1-diyl))bis(oxy))(11aS,11a'S)-bis(11-hydroxy-7-methoxy-2-methylene-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate)1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate (1-21g)

[0471] DMP (55.27 mg, 0.13 mmol) was added at 0°C to a solution of 1-21f (52 mg, 0.059 mmol) in CH2Cl2 (2 mL). The reaction mixture was then warmed to room temperature and stirred for 4 hours. The reaction mixture was filtered, and the filtrate was quenched with saturated sodium thiosulfate aqueous solution (5 mL). Subsequently, saturated NaHCO3 aqueous solution (5 mL) and H2O (10 mL) were slowly added. The mixture was extracted with dichloromethane (5 mL x 3), the organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated to obtain the crude product. This was purified by flash column chromatography (eluate: CH2Cl2 / MeOH = 20 / 1) to obtain 1-21 g (32 mg, yield 61.82%) as a white solid.

[0472] MS(ESI)m / z:874.4[M+H] + .

[0473] Step 7: (11aS,11a'S)-8,8'-((Azandiylbis(ethane-2,1-diyl))bis(oxy))bis(7-methoxy-2-methylene-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5-one)(1,21)

[0474] To a solution of 1-21 g (32 mg, 0.036 mmol) and CH2Cl2 (2 mL), Pd(PPh3)4 (2.12 mg, 0.0018 mmol) and pyrrolidine (6 μL, 0.073 mmol) were added, and the mixture was stirred at room temperature under N2 for 15 minutes. The reaction product was neutralized with AcOH, concentrated to obtain the crude product, which was purified by preparative HPLC (column: XBridge Prep C18 OBD 5 μm 19*150 mm; mobile phase: A-water (0.01% formic acid): B-acetonitrile; flow rate: 20 mL / min) to obtain 1-21 (9.7 mg, yield 45.23%) as a white solid.

[0475] MS(ESI)m / z:586.3[M+H] + .

[0476] Examples 1-22 [ka] Step 1: 2-Bromo-N-(2-bromoethyl)-N-methylethane-1-amine(1-22b)

[0477] To a solution of 37% formaldehyde (0.66 mL) and 98% formic acid (0.26 mL), 1-22a (1.0 g) was added, and the solution was heated under reflux for 2 hours. The mixture was concentrated under vacuum to obtain a colorless oil, which was crystallized with MeOH to obtain 1-22b (700 mg, yield 65%) as a white solid.

[0478] 1 H NMR (400MHz, DMSO-d6) δ9.66 (s, 1H), 3.79-3.72 (m, 4H), 3.65-3.58 (m, 4H), 2.85 (s, 3H).

[0479] Step 2: ((((Methylazanediyl)bis(ethane-2,1-diyl))bis(oxy))bis(5-methoxy-2-nitro-4,1-phenylene))bis(((S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-methylenepyrrolidine-1-yl)methanone)(1-22c)

[0480] To a solution of 1-22b (100 mg, 0.408 mmol) and 1-21b (379.50 mg, 0.898 mmol) in DMSO (3 mL), K2CO3 (112.84 mg, 0.816 mmol) was added. The mixture was stirred at 50°C for 16 hours. The reaction product was quenched with H2O and extracted with siRNA (30 mL). The organic phase was dried over Na2SO4, filtered, and concentrated to obtain the crude product, which was purified by silica column gel chromatography (eluate: hexane / siRNA = 100 / 0~0 / 100) to obtain 1-22c (200 mg, yield 52.78%) as a yellow solid.

[0481] MS(ESI)m / z:928.6[M+H] + .

[0482] Step 3: (((methylazandiyl)bis(ethane-2,1-diyl))bis(oxy))bis(5-methoxy-2-nitro-4,1-phenylene))bis(((S)-2-(hydroxymethyl)-4-methylenepyrrolidine-1-yl)methanone)(1-22d)

[0483] To a solution of 1-22c (200 mg, 0.215 mmol) in THF (2 mL) and water (0.1 mL), p-toluenesulfonic acid hydrate (90.16 mg, 0.474 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The mixture was concentrated and purified by silica gel chromatography (eluate: CH2Cl2 / MeOH = 20 / 1) to obtain 1-22d (110 mg, yield 72.96%) as a white solid.

[0484] MS(ESI)m / z:700.4[M+H] + .

[0485] Step 4: (2S,2'S)-1,1'-(4,4'-(((methylazanediyl)bis(ethane-2,1-diyl))bis(oxy))bis(5-methoxy-2-nitrobenzoyl))bis(4-methylenepyrrolidine-2-carbaldehyde)(1-22e)

[0486] DMP (146.69 mg, 0.345 mmol) was added at 0°C to a solution of 1-22d (110 mg, 0.157 mmol) in dichloromethane (2 mL), THF (2 mL), and DMF (1 mL). The reaction mixture was then warmed to room temperature and stirred at room temperature for 4 hours. The reaction mixture was filtered, and the filtrate was concentrated to obtain the crude product, which was purified by silica gel chromatography (eluate: CH2Cl2 / MeOH = 20 / 1) to obtain 1-22e (60 mg, yield 55.83%) as a yellow solid.

[0487] MS(ESI)m / z:696.3[M+H] + .

[0488] Step 5: (11aS,11a'S)-8,8'-(((methylazandiyl)bis(ethane-2,1-diyl))bis(oxy))bis(7-methoxy-2-methylene-1,2,3,11a-tetrahydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5-one)(1,22)

[0489] To a solution of 1-22e (60 mg, 0.086 mmol) in THF (0.896 mL), methanol (4.48 mL), and water (0.896 mL), NH4Cl (92.26 mg, 1.72 mmol) was added, followed by iron powder (48.16 mg, 0.862 mmol). The mixture was then heated under N2 at 50°C for 16 hours. The reaction mixture was cooled to room temperature and filtered through Celite. The filtrate was concentrated to obtain the crude product, which was purified by preparative HPLC (column: XBridge Prep C18 OBD 5 μm 19*150 mm; method: mobile phase: A-water (0.01% formic acid): B-acetonitrile; flow rate: 20 mL / min) to obtain 1-22 (4.8 mg, yield 9.28%) as a white solid.

[0490] MS(ESI)m / z:600.3[M+H] + .

[0491] 1 H NMR(400MHz,CDCl3)δ7.68(d,J=4.4Hz,2H),7.49(s,2H),6.83(s,2H),5.19(d,J=11.2Hz,4H),4.3 8-4.17(m,8H),3.91(s,6H),3.89-3.84(m,2H),3.12-3.08(m,6H),2.97-2.93(m,2H),2.56(s,3H).

[0492] Example 2-1 [ka] Step 1: Allyl((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate(2-1c)

[0493] EEDQ (2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline, 4.77 g, 19.3 mmol) was added to a solution of 2-1a (5 g, 18.4 mmol) and 2-1b (2.37 g, 19.3 mmol) in 100 mL of dry THF. The mixture was stirred at room temperature for 40 hours. After the reaction was complete, as determined by LC-MS / TLC (CH2Cl2 / MeOH = 20:1), the mixture was concentrated. The residue was slurryed with MTBE (30V) and stirred for 2 hours. The solid was isolated by filtration under vacuum for 3 hours to obtain 2-1c (5.16 g, 74% yield). MS(ESI) m / z: 378.4 [M+H] + .

[0494] 1 H NMR(400MHz,d6-DMSO)δ9.90(s,1H),8.14(d,J=7.0Hz,1H),7.53(d,J=8.5Hz,2H),7.25(t,J=9.0 Hz,3H),5.91(ddd,J=22.3,10.5,5.3Hz,1H),5.30(dd,J=17.2,1.5Hz,1H),5.17(d,J=10.5Hz,1H) ),5.10(t,J=5.7Hz,1H),4.52-4.45(m,2H),4.43(d,J=5.6Hz,3H),3.89(dd,J=8.5,7.1Hz,1H),1 .98(dq,J=13.5,6.7Hz,1H),1.28(t,J=11.8Hz,3H),0.88(d,J=6.8Hz,3H),0.84(d,J=6.7Hz,3H).

[0495] Step 2: (S)-2-amino-N-((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide(2-1d)

[0496] Pd(PPh3)4 (76.6 mg, 0.066 mmol) was added to a solution of 2-1c (500 mg, 1.33 mmol) in CH2Cl2 (10 mL) and pyrrolidine (270.8 μL, 3.31 mmol) under N2 conditions at room temperature. The reaction mixture was stirred at room temperature for 0.5 hours. The reaction product was concentrated and purified by silica column gel chromatography (CH2Cl2 / MeOH = 90:10) to obtain product 2-1d (370 mg, yield 95%) as a white solid.

[0497] MS(ESI)m / z:294.3[M+H] + .

[0498] 1 H NMR(400MHz,d6-DMSO)δ9.99(s,1H),8.20(d,J=6.3Hz,1H),7.53(d,J=8.4Hz,2H),7.24(d,J=8.4Hz,2H),5.10(s,1H),4.53-4.45(m,1H) ),4.43(s,2H),3.05(d,J=4.9Hz,1H),1.93(dd,J=12.0,6.8Hz,1H),1.30(d,J=7.0Hz,3H),0.89(d,J=6.9Hz,3H),0.80(d,J=6.8Hz,3H).

[0499] Step 3: (9H-Fluoren-9-yl)methyl((17S,20S)-21-((4-(hydroxymethyl)phenyl)amino)-17-isopropyl-20-methyl-15,18,21-trioxo-3,6,9,12-tetraoxa-16,19-diazahenicosyl)carbamate(2-1f)

[0500] DIPEA (326 mg, 2.52 mmol) was added to a solution of 2-1e (370 mg, 1.26 mmol) and HATU (575.8 mg, 1.51 mmol) in 4 mL of dry DMF. The mixture was stirred at room temperature for 10 minutes. Then, a solution of 2-1d (645 mg, 1.33 mmol) in DMF was added to the mixture. The reaction was stirred for 1 hour. After the reaction was completed, as determined by LC-MS / TLC (CH2Cl2 / MeOH=20:1), the mixture was concentrated and purified by silica column gel chromatography (CH2Cl2 / MeOH=95:5) to obtain product 2-1e (680 mg, yield 71%) as a light brown solid. MS(ESI)m / z:763.5[M+H] + .

[0501] Step 4: tert-butyl(5-((5-(5-((((4-((21S,24S)-1-(9H-fluoren-9-yl)-21-isopropyl-24-methyl-3,19,22-trioxo-2,7,10,13,16-pentaoxa-4,20,23-triazapentacosan-25-amide)benzyl)oxy)carbonyl)amino)-4-((S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-methylenepyrrolidine-1-carbonyl)-2-methoxyphenoxy)pentyl)oxy)-2((S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-methylenepyrrolidine-1-carbonyl)-4-methoxyphenyl)carbamate(2-1h)

[0502] To a 4Å MS solution of 2-1g (200mg, 0.121mmol) and 200mg in THF (2.5mL), triphosgene (24.9mg, 0.084mmol) was added under N2 at 0°C, followed by TEA (64μL, 0.462mmol). The mixture was stirred for 10 minutes under N2 at 0°C. Isocyanate formation was monitored by LCMS analysis after quenching with methanol. A solution of 2-1f (176mg, 0.231mmol), dibutyltin dilaurate (13.3mg, 0.021mmol), and TEA (43.7μL, 0.315mmol) in THF (2.5mL) was added to the mixture. The mixture was stirred at room temperature for 3 hours. The mixture was filtered, and the filter was concentrated. The residue was purified by silica column gel chromatography (CH2Cl2 / MeOH=96:4) to obtain product 2-1h (302 mg, yield 83%) as a white solid. MS(ESI)m / z:1472.3[M+H] + .

[0503] Step 5: tert-butyl(5-((5-(5-((((4-((21S,24S)-1-(9H-fluoren-9-yl)-21-isopropyl-24-methyl-3,19,22-trioxo-2,7,10,13,16-pentaoxa-4,20,23-triazapentacosan-25-amide)benzyl)oxy)carbonyl)amino)-4-((S)-2-(hydroxymethyl)-4-methylenepyrrolidine-1-carbonyl)-2-methoxyphenoxy)pentyl)oxy)-2-((S)-2-(hydroxymethyl)-4-methylenepyrrolidine-1-carbonyl)-4-methoxyphenyl)carbamate(2-1i)

[0504] To a solution of 2-1h (304 mg, 0.17 mmol) in THF (3 mL) and water (0.15 mL), p-toluenesulfonic acid hydrate (40 mg, 0.21 mmol) was added. The reaction mixture was stirred at 22°C for 4 hours. After determining the completion of the reaction by TLC (CH2Cl2 / MeOH=20:1), the mixture was diluted with  (20 mL) and washed with water, saturated NaHCO3, and brine. The organic phase was concentrated and purified by silica column gel chromatography (CH2Cl2 / MeOH=95:5) to obtain product 2-1i (213 mg, yield 81%). MS(ESI) m / z: 1514.1[M+H] + .

[0505] Step 6: 4-((21S,24S)-1-(9H-fluoren-9-yl)-21-isopropyl-24-methyl-3,19,22-trioxo-2,7,10,13,16-pentaoxa-4,20,23-triazapentacosan-25-amide)benzyl(11S,11aS)-8-((5-(((11S,11aS)-10-(tert-butoxycarbonyl)-11-hydroxy-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)-11-hydroxy-7-methoxy-2-methylene-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(2-1j)

[0506] DMP (58.9 mg, 0.139 mmol) was slowly and gradually added to a solution of 2-1i (100 mg) in dry CH2Cl2 (2 mL) at 0°C. The reaction mixture was then warmed to room temperature and stirred overnight. The reaction mixture was quenched with saturated Na2S2O3, followed by the addition of saturated NaHCO3 and water. The layers were separated, and the organic layer was washed with saturated Na2S2O3, saturated NaHCO3, and brine, and dried over Na2SO4. The crude product was purified by silica column gel chromatography (CH2Cl2 / MeOH = 95 / 5) to obtain product 2-1j (80 mg, yield 80%). MS(ESI) m / z: 1510.3[M+H] + .

[0507] Step 7: 4-((21S,24S)-1-(9H-fluoren-9-yl)-21-isopropyl-24-methyl-3,19,22-trioxo-2,7,10,13,16-pentaoxa-4,20,23-triazopentacosan-25-amide)benzyl(11S,11aS)-11-hydroxy-7-methoxy-8-((5-(((S)-7-methoxy C-2-methylene-5-oxo-2,3,5,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)-2-methylene-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(2-1k)

[0508] 2-1k (60 mg, 0.040 mmol) was cooled to -3°C. Separately, a 95% TFA solution in H2O (1.5 mL) was cooled to -3°C and added to 2-1k. The reaction mixture was stirred at -3°C for 40 minutes and then poured into a 1:1 solution of CHCl3 / NaHCO3 (40 mL) at 0°C. The organic layer was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The crude material 2-1k was used directly in the next step.

[0509] MS(ESI)m / z:1392.6[M+H] + .

[0510] Step 8: 4-((17S,20S)-1-amino-17-isopropyl-20-methyl-15,18-dioxo-3,6,9,12-tetraoxa-16,19-diazahenicosan-21-amide)benzyl(11S,11As)-11-hydroxy-7-methoxy-8-((5-(((S)-7-methoxy-2-methylene-5-oxo-2,3,5,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)2-methylene-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(2-1l)

[0511] To a crude 2-1k solution in 0.5 mL of DMF, Et2NH (61 μL, 0.593 mmol) was added. The mixture was stirred at room temperature for 0.5 hours. After the reaction was complete, the mixture was purified by preparative HPLC (0.1% FA in H2O) to obtain 2-1 L of product (14.3 mg, yield 31%, 2 steps).

[0512] MS(ESI)m / z:1170.3[M+H] + .

[0513] Step 9: 4-((21S,24S)-1-((1R,8S,9s)-Bicyclo[6.1.0]nonano-4-in-9-yl)-21-isopropyl-24-methyl-3,19,22-trioxo-2,7,10,13,16-pentaoxa-4,20,23-triazopentacosan-25-amide)benzyl(11S,11As)-11-hydroxy-7-methoxy-8-((5- (((S)-7-methoxy-2-methylene-5-oxo-2,3,5,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentyl)oxy)2-methylene-5-oxo-2,3,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-10(5H)-carboxylate(2-1)

[0514] DIPEA (8.6 μL, 0.05 mmol) was added to solutions of 2-1 l (14.5 mg, 0.012 mmol) and 2-1 m (10.8 mg, 0.037 mmol) in 1 mL of DMF. The mixture was stirred at room temperature for 20 minutes. The mixture was purified by preparative HPLC (without additives in the mobile phase) to obtain product 2-1 (7.7 mg, 46% yield) as a pale gray solid.

[0515] MS(ESI)m / z:1346.4[M+H] + .

[0516] After linker cleavage, the payload released from linker-payload 2-1 undergoes a dehydration reaction to form SG-2057, a compound with similar potency to payload Ref-1-1 (also known as SG-3199).

[0517] Example 2-2 [ka] [ka] Step 1: Allyl(S)-7-methoxy-5-oxo-8-((triisopropylsilyl)oxy)-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(2-2a)

[0518] DIAD (1.14 mL, 5.75 mmol) was added to a solution of 1-7c (1.53 g, 2.87 mmol) and PPh3 (2.26 g, 8.62 mmol) in THF (30 mL). The reaction mixture was stirred at 40°C under N2 for 1 hour. After the reaction was complete by TLC (petroleum ether / Depositphotos = 1:2), the solvent was removed under vacuum, and the residue was purified by silica column gel chromatography to obtain the product (petroleum ether / Depositphotos = 40:60) 2-2a (940 mg, yield 64%). MS (ESI) m / z: 515.5 [M + H] + .

[0519] 1 H NMR(400MHz,CDCl3)δ7.19(s,1H),6.70(s,1H),5.84-5.68(m,1H),5.12(t,J=12.7Hz,2H),4.58(d d,J=12.9,5.3Hz,1H),4.45(t,J=12.5Hz,2H),3.89-3.86(m,1H),3.85(s,3H),3.74(d,J=11.8Hz, 1H),3.48(dd,J=12.2,4.2Hz,1H),3.37(d,J=11.8Hz,1H),2.37(dd,J=12.8,8.4Hz,1H),1.42(d,J =12.8Hz,1H),1.25(ddd,J=19.0,9.0,4.5Hz,3H),1.08(dd,J=7.3,3.7Hz,18H),0.86-0.57(m,4H).

[0520] Step 2: Allyl(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)-carboxylate(2-2b)

[0521] Lithium acetate (121 mg, 1.83 mmol) was added to a solution of 2-2a (940 mg, 1.83 mmol) in wet DMF (15 mL of 49 / 1 DMF / water). The reaction was allowed to proceed at 25°C for 2 hours. The mixture was diluted with siRNA and washed twice with H2O and brine. The organic phase was dried over Na2SO4, concentrated, and purified by silica column gel chromatography to obtain product 2-2b (605 mg, 92% yield). MS(ESI) m / z: 359.4 [M+H] + .

[0522] Step 3: (S)-(3-(hydroxymethyl)-3,4-dihydroisoquinoline-2(1H)-yl)(5-methoxy-2-nitro-4-((triisopropylsilyl)oxy)phenyl)methanone(2-2e)

[0523] EDCI (935 mg, 4.88 mmol) was added at 0°C to a solution of 2-2c (1.5 g, 4.06 mmol) and 2-pyridinol 1-oxide (HOPO, 497 mg, 4.47 mmol) in CH2Cl2 (15 mL). The reaction was allowed to proceed at 15°C for 1 hour, at which point a solution of 2-2d (729 mg, 4.47 mmol) and triethylamine (0.71 mL, 5.08 mmol) in CH2Cl2 (15 mL) was added at -10°C. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was washed with water (approximately 30 mL), followed by cold aqueous HCl (0.5 M), until the pH was adjusted to 4-5. The organic phase was then washed with saturated aqueous NaHCO3 (approximately 30 mL), followed by water (approximately 30 mL). The solvent was removed under vacuum to obtain the crude product, which was then purified by silica column gel chromatography to obtain product 2-2e (1.34 g, yield 57%). MS(ESI)m / z: 515.4[M+H] + .

[0524] Step 4: (S)-(3-(((tert-butyldimethylsilyl)oxy)methyl)-3,4-dihydroisoquinoline-2(1H)-yl)(5-methoxy-2-nitro-4-((triisopropylsilyl)oxy)phenyl)methanone(2-2f)

[0525] Imidazole (355 mg, 5.21 mmol) was added to a solution of 2-2e (1.34 g, 2.61 mmol) in 25 mL of CH2Cl2. Then, TBSCl (589 mg, 3.91 mmol) was added to the mixture at room temperature. The mixture was stirred overnight at room temperature. The reaction product was filtered, and the filtrate was concentrated. The crude product was purified by silica column gel chromatography to obtain product 2-2f (1.38 g, yield 84%). MS(ESI) m / z: 629.5[M+H] + .

[0526] Step 5: (S)-(3-(((tert-butyldimethylsilyl)oxy)methyl)-3,4-dihydroisoquinoline-2(1H)-yl)(4-hydroxy-5-methoxy-2-nitrophenyl)methanone (2-2g)

[0527] Lithium acetate (145 mg, 2.2 mmol) was added to a solution of 2-2 f (1.38 g, 2.2 mmol) in wet DMF (15 mL, 49 / 1 DMF / water). The reaction was allowed to proceed at 25°C for 2 hours. The mixture was diluted with siRNA and washed twice with H2O and brine. The organic phase was concentrated and purified by silica column gel chromatography to obtain 2-2 g (930 mg, 90% yield) of the product. MS(ESI) m / z: 473.3 [M+H] + .

[0528] Step 6: (S)-(4-((5-bromopentyl)oxy)-5-methoxy-2-nitrophenyl)(3-(((tert-butyldimethylsilyl)oxy)methyl)-3,4-dihydroisoquinoline-2(1H)-yl)methanone(2-2h)

[0529] To a solution of 2-2 g (930 mg, 1.97 mmol) and 1,5-dibromopentane (4.0 mL, 29.5 mmol) in 18 mL of DMF, K2CO3 (330 mg, 2.36 mmol) was added at room temperature. The mixture was stirred at room temperature for 2 hours. The mixture was diluted with siRNA and washed with H2O and brine. The organic phase was concentrated and purified by silica column gel chromatography (petroleum ether / siRNA = 75 / 25) to obtain product 2-2 h (818 mg, yield 67%). MS(ESI) m / z: 621.4 [M+H] + .

[0530] Step 7: Allyl(S)-8-((5-(4-((S)-3-(((tert-butyldimethylsilyl)oxy)methyl)-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)-2-methoxy-5-nitrophenoxy)pentyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(2-2i)

[0531] 2-2b (450 mg, 1.26 mmol) and K2CO3 (226 mg, 1.63 mmol) were added to a solution of 2-2h (818 mg, 1.32 mmol) in DMF (1 mL). The mixture was stirred at room temperature for 24 hours. The product (petroleum ether / siRNA = 1:2) was detected by LC-MS. The mixture was diluted with siRNA and washed with water and brine. The organic phase was concentrated and purified by silica column gel chromatography (petroleum ether / siRNA = 33:67) to obtain product 2-2i (930 mg, yield 82%). MS(ESI) m / z: 899.6[M+H] + .

[0532] Step 8: Allyl(S)-8-((5-(5-amino-4-((S)-3-(((tert-butyldimethylsilyl)oxy)methyl)-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)-2-methoxyphenoxy)pentyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(2-2j)

[0533] Zinc powder (2.65 g, 38.3 mmol) was added at 0°C to a mixture of ethanol (10 mL), water (0.625 mL), and AcOH (0.625 mL). The reaction mixture was stirred at 5°C for 30 minutes. A solution of 2-2i (930 mg, 1.04 mmol) in ethanol (6 mL) was added dropwise at 5°C. The reaction was allowed to proceed at 5°C for 50 minutes. The solid was removed by filtration. The filtrate was diluted with  and washed with water, saturated NaHCO3 aqueous solution, and brine. The organic phase was dried over sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure to obtain the crude product. This was purified by silica column gel chromatography (petroleum ether / Â=33:67) to obtain product 2-2j (806 mg, 90% yield) as a yellow solid. MS(ESI)m / z:868.7[M+H] + .

[0534] Step 9: Allyl(S)-8-((5-(5-((((4-((21S,24S)-1-(9H-Fluoren-9-yl)-21-Isopropyl-24-methyl-3,19,22-Trioxo-2,7,10,13,16-Pentaoxa-4,20,23-Triazapentacosan-25-amide)Benzyl)Oxy)Carbonyl)Amino)-4-((S)-3-(((tert -Butyldimethylsilyl)oxy)methyl)-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)-2-methoxyphenoxy)pentyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(2-2l)

[0535] 2-2L was prepared according to the procedure described in step 4 of Example 2-1 to obtain a white solid (260 mg, 91% yield). MS(ESI)m / z:1658.4[M+H] + .

[0536] Step 10: Allyl(S)-8-((5-(5-((((4-((21S,24S)-1-(9H-Fluoren-9-yl)-21-Isopropyl-24-methyl-3,19,22-Trioxo-2,7,10,13,16-Pentaoxa-4,20,23-Triazapentacosan-25-amide)Benzyl)Oxy)Carbonyl)Amino)-4-((S)- 3-(hydroxymethyl)-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)-2-methoxyphenoxy)pentyl)oxy)-7-methoxy-5-oxo-11,11a-dihydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-10(5H)-carboxylate(2-2m)

[0537] To a solution of 2-2 L (260 mg, 0.16 mmol) of para-toluenesulfonic acid hydrate (29.8 mg, 0.16 mmol) in THF (6 mL) and water (0.3 mL), p-toluenesulfonic acid hydrate (29.8 mg, 0.16 mmol) was added. The reaction mixture was stirred overnight at 22 °C. The mixture was diluted with  (20 mL) and washed with water and brine. The organic phase was concentrated and purified by silica column gel chromatography to obtain product 2-2 m (172 mg, yield 71%). MS(ESI) m / z: 1544.3 [M+H] + .

[0538] Step 11: 4-((21S,24S)-1-(9H-fluoren-9-yl)-21-isopropyl-24-methyl-3,19,22-trioxo-2,7,10,13,16-pentaoxa-4,20,23-triazapentacosan-25-amide)benzyl(6S,6As)-3-((5-(((S)-10-((allyloxy)carbonyl)-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)6-hydroxy-2-methoxy-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(2-2n)

[0539] DMP (52 mg, 0.123 mmol) was slowly and gradually added to a solution of 2-2m (172 mg) in 2.5 mL of dry CH2Cl2 at 0°C. The reaction mixture was then warmed to room temperature and stirred for 9 hours. After 9 hours, the reaction mixture was quenched with saturated Na2S2O3, followed by the addition of saturated NaHCO3 and water. The layers were separated, and the organic layer was washed with saturated Na2S2O3, saturated NaHCO3, and brine, and dried over Na2SO4. The crude product was purified by silica column gel chromatography (CH2Cl2 / MeOH = 95 / 5) to obtain product 2-2n (130 mg, yield 76%). MS(ESI) m / z: 1542.1[M+H] + .

[0540] Step 12: 4-((17S,20S)-1-amino-17-isopropyl-20-methyl-15,18-dioxo-3,6,9,12-tetraoxa-16,19-diazahenicosan-21-amide)benzyl(6S,6As)-3-((5-(((S)-10-((allyloxy)carbonyl)-7-methoxy-5-oxo-5,10,11,11a-tetra Hydro-1H,3H-spiro[benzo[e]pyrrolo[1,2-a][1,4]diazepine-2,1'-cyclopropane]-8-yl)oxy)pentyl)oxy)6-hydroxy-2-methoxy-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(2-2o)

[0541] To a solution of 2-2n (50 mg, 0.04 mmol) in DMF (2 mL), Et2NH (38 μL, 0.37 mmol) was added. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the mixture was concentrated to obtain the crude product 2-2o, which was used directly in the next step. MS(ESI)m / z:1320.2[M+H] + .

[0542] Step 13: 4-((17S,20S)-1-amino-17-isopropyl-20-methyl-15,18-dioxo-3,6,9,12-tetraoxa-16,19-diazahenicosan-21-amide)benzyl(6S,6aS)-6-hydroxy-2-methoxy-3-((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)-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(2-2p)

[0543] Pd(PPh3)4 (2.15 mg, 0.002 mmol) was added at room temperature to a solution of crude 2-2o in THF / MeOH (2 mL / 0.2 mL) and dimedone (10.42 mg, 0.074 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction product was concentrated. The residue was purified by preparative HPLC (0.1% FA in H2O) to obtain product 2-2p (16 mg, yield 61%) as a white solid. MS(ESI) m / z: 1236.1[M+H] + .

[0544] Step 14: 4-((21S,24S)-1-((1R,8S,9s)-Bicyclo[6.1.0]nonano-4-in-9-yl)-21-isopropyl-24-methyl-3,19,22-trioxo-2,7,10,13,16-pentaoxa-4,20,23-triazapentacosan-25-amide)benzyl(6S,6As)-6-hydroxy-2-methoxy-3-((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)-14-oxo-6,6a,7,12-tetrahydrobenzo[5,6][1,4]diazepino[1,2-b]isoquinoline-5(14H)-carboxylate(2-2)

[0545] DIPEA (10.9 μL, 0.063 mmol) was added to a solution of 2-2p (19.3 mg, 0.016 mmol) and 2-1m (13.6 mg, 0.047 mmol) in DMF (1 mL). The mixture was stirred at room temperature for 20 minutes. The mixture was purified by preparative HPLC (0.1% FA in H2O) to obtain product 2-2 (9.8 mg, yield 44%) as a white solid. MS(ESI) m / z: 1412.4[M+H] + .

[0546] After linker cleavage, the payload released from linker-payload 2-2 undergoes a dehydration reaction to form the compounds of Examples 1-3.

[0547] The compounds disclosed herein may also be synthesized based on the synthetic methods provided herein, in combination with general knowledge of the art. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 2]

[0548] Preparation and Characterization of ADCs Preparation of DAR2 antibody drug conjugates. Anti-CD74 antibody mAb1 (concentration 0.5-25 mg / mL, 50 mM Tris-HCl buffer, pH 7.0-8.5) in reaction buffer was incubated with endoS2 at reaction temperature (0-40°C) for 1-24 hours at a concentration of 1 / 2000-1 / 500 w / w (EndoS2 / mAb weight ratio) for 1-24 hours. 2-40 equivalents of UDP-GalNAz (20 mM) and β1,4-GalT at a concentration of 0.1 w / w%-10 w / w% (GalT / mAb weight ratio) were added to the reaction mixture and incubated at reaction temperature (0-40°C) for 8-24 hours in reaction buffer (50 mM Tris-HCl buffer, pH 7.0-8.5, 20 mM MnCl2). The reaction mixture was purified with protein A resin to obtain mAb1-GalNAz.

[0549] Organic solvents (e.g., DMSO, DMF, DMA, PG, acetonitrile, 0-25% v / v) and linker-payload stock (10-25 equivalents, 10 mM stock in organic solvent) were added stepwise to reaction buffer (PBS buffer, pH 7.0-8.5) together with mAb1-GalNAz (1-20 mg / mL) at 0-25°C for 0.5-24 hours. The solutions were subjected to buffer exchange (spin desalination column, ultrafiltration, and dialysis) to storage buffer (e.g., histidine acetate buffer, pH 5.5-6.5, with optional additives such as sucrose, trehalose, and Tween® 20, 60, 80).

[0550] ADC Characterization. The ADCs were characterized using the following analytical methods. The SEC purity of all ADCs was >95%.

[0551] Determination of drug-to-antibody ratio (DAR) by LCMS or HIC method LC-MS method: LC-MS analysis was performed under the following measurement conditions. LC-MS system: Vanquish Flex UHPLC and Orbitrap Exploris 240 mass spectrometer Column: MAbPac(TM) RP, 2.1*50mm, 4μm, 1,500Å, Thermo Scientific(TM) Column temperature: 80℃ Mobile phase A: 0.1% formic acid (FA) aqueous solution Mobile phase B: Acetonitrile solution containing 0.1% formic acid (FA) Gradient Programs: 25%B~25%B (0 min~2 min), 25%B~50%B (2 min~18 min), 50%B~90%B (18 min~18.1 min), 90%B~90%B (18.1 min~20 min), 90%B~25%B (20 min~20.1 min), 25%B~25%B (20.1 min~25 min) Amount of sample injected: 1 μg MS parameters: Intact and denatured MS data were acquired in HMR mode with R=15k and deconvolved using the ReSpect algorithm and sliding window integration with Thermo Scientific® BioPharma Finder® 4.0 software. HIC method: HPLC analysis was performed under the following measurement conditions. HPLC System: Waters ACQUITY ARC HPLC System Detector: Measurement wavelength: 280nm Column: Tosoh Bioscience 4.6μm ID×3.5cm, 2.5μm butyl nonporous resin column Column temperature: 25℃ Mobile phase A: 1.5M ammonium sulfate, 50mM phosphate buffer, pH 7.0 Mobile phase B: 50 mM phosphate buffer, 25% (V / V) isopropanol, pH 7.0 Gradient Programs: 0% B~0% B (0 min~2 min), 0% B~100% B (2 min~15 min), 100% B~100% B (15 min~16 min), 100% B~0% B (16 min~17 min), 0% B~0% B (17 min~20 min) Amount of sample injected: 20 μg

[0552] SEC method for determining the purity of ADCs

[0553] HPLC analysis was performed under the following measurement conditions: HPLC System: Waters H-Class UPLC System Detector: Measurement wavelength: 280nm Column: ACQUITY UPLC BEH200 SEC 1.7um 4.6x150mm, Waters Column temperature: Room temperature Mobile phase A: 200 mM phosphate buffer, 250 mM potassium chloride, 15% isopropyl alcohol, pH 7.0 Gradient program: 10 minutes of isocratic elution, flow rate 0.3 mL / min Amount of sample injected: 20 μg

[0554] ADC hydrophobicity evaluation by HIC method: More hydrophobic ADCs exhibit slower retention times in HIC (hydrophobic interaction column) chromatography. The DAR2 peak was used as a reference.

[0555] HPLC analysis was performed under the following measurement conditions. Method 1 HPLC System: Waters ACQUITY ARC HPLC System Detector: Measurement wavelength: 280nm Column: Tosoh Bioscience 4.6μm ID×3.5cm, 2.5μm butyl nonporous resin column Column temperature: 25℃ Mobile phase A: 1.5M ammonium sulfate, 50mM phosphate buffer, pH 7.0 Mobile phase B: 50 mM phosphate buffer, 25% (v / v) isopropanol, pH 7.0 Gradient Programs: 0%B~0%B (0 min~2 min), 0%B~100%B (2 min~15 min), 100%B~100%B (15 min~16 min), 100%B~0%B (16 min~17 min), 0%B~0%B (17 min~20 min) Amount of sample injected: 20 μg Method 2 HPLC System: Waters ACQUITY ARC HPLC System Detector: Measurement wavelength: 280nm Column: MABPac HIC-10, 5μm, 4.6×10mm (Thermo) Column temperature: 25℃ Mobile phase A: 1.5M ammonium sulfate, 50mM sodium phosphate, pH 7.0 Mobile phase B: 50 mM sodium phosphate, pH 7.0 Gradient program: 20%B~20%B (0 min~1 min), 0%B~0%B (1 min~35 min), 20%B~20%B (35 min~40 min) Flow rate: 0.5mL / min Sample preparation: The sample was diluted with the initial mobile phase to a concentration of 0.5 mg / mL. [Table 3]

[0556] Anti-CD74 antibody mAb1 Light chain sequence (SEQ ID NO: 1)

[0557] DIQMTQSPSSVSASVGDRVTITCRASQGIGSWLAWYQQKPGKAPKLLIYAADRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYHTYPLTFGGGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0558] Heavy chain sequence (SEQ ID NO: 2)

[0559] QVQLVESGGGVVQPGRSLRLSCAASGFNFSDYGMHWVRQAPGKGLEWVAVIWYDGSISYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGTVEHGAVYGTDVW GQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0560] cell line A375 (ATCC, CRL-1619). A375 is an epithelial cell line isolated from the skin of a 54-year-old female patient with malignant melanoma, and was purchased from ATCC. The basic medium for A375 was GlutaMAX® Supplement (Gibco, 10566024), a high-glucose DMEM. To prepare a full-growth medium, fetal bovine serum was added to the basic medium to a final concentration of 10% (Gibco, 10099-141C). The cell line was grown at 37°C in a humidified 5% CO2 atmosphere, and the presence or absence of mycoplasma was checked periodically using the MycoAlert® PLUS Mycoplasma Detection Kit (Lonza, LT07-710).

[0561] Calu-6 (ATCC, HTB-56). Calu-6 is an epithelial cell line isolated from a Caucasian female patient with undifferentiated carcinoma, and was purchased from ATCC. The basic medium for Calu-6 was Eagle's Minimal Essential Medium (ATCC, 30-2003). To prepare a full-growth medium, fetal bovine serum was added to the basic medium to a final concentration of 10% (Gibco, 10099-141C). The cell line was grown at 37°C in a humidified 5% CO2 atmosphere, and the presence or absence of mycoplasma was checked periodically using the MycoAlert® PLUS Mycoplasma Detection Kit (Lonza, LT07-710).

[0562] Compound cell cytotoxicity in A375 and Calu-6 cancer strains Direct killing of the payload by compounds 1-1 to 1-22 was evaluated in A375 and Calu-6 cancer cell lines. Cells were seeded in 96-well plates (Greiner: 655090) at 100 μL / well, with 1E3 cells per well (A375) and 2E3 cells per well (Calu-6), and incubated overnight at 37°C and 5% CO2. Fresh growth medium containing various concentrations of compounds was added at 50 μL / well, and incubated at 37°C and 5% CO2 for 6 days. Cell viability was detected at 70 μL / well using Cell Titer-Glo (Promega, G7573). The plates were incubated at room temperature for 10 minutes to stabilize the luminescence signal. The plates were analyzed using a microplate reader.

[0563] The cell killing data for compounds 1-1 to 1-5 are shown in Figures 1 and 2, and in the table below. [Table 4]

[0564] The cell killing data for compounds 1-6 to 1-10 are shown in Figures 3 and 4, and in the table below. [Table 5]

[0565] The cell killing data for compounds 1-11 to 1-13 are shown in Figures 5 and 6, and in the table below. [Table 6]

[0566] The cell killing data for compounds 1-14, 1-15, 1-17, and 1-20 are shown in Figures 7 and 8, and in the table below. [Table 7]

[0567] The cell killing data for compounds 1-16 and 1-19 are shown in Figures 9 and 10, and in the table below. [Table 8]

[0568] The cell killing data for compounds 1-18 are shown in Figures 11 and 12, and in the table below. [Table 9]

[0569] The cell killing data for compounds 1-21 and 1-22 are shown in Figures 13 and 14, and in the table below. [Table 10]

[0570] While the above disclosure has been presented in some detail with illustrations and examples for clear understanding, it will be apparent to those skilled in the art that certain minor changes and modifications will be made. Therefore, the description and examples should not be construed as limiting.

[0571] Where any prior art publication is referenced herein, it should be understood that such reference does not constitute an endorsement that such publication forms part of the common general knowledge in the art in any country.

[0572] All non-patent literature, patents, patent applications, and published patent application disclosures referenced herein by identifying citation are incorporated herein by reference in their entirety.

Claims

1. Compound of formula (I): 【Chemistry 81】 or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, In the formula, each of ring A and ring B is independently one of the following formulas: 【Chemistry 82-1】 【Chemistry 82-2】 【Chemistry 83】 This indicates the linking point to the linker. The linker is -(CH ２ ), -(CH ｒ ), -(CH ２ ), -X-(CH ｐ ), -(CH ２ ), -(CH ｑ ), or -(CH ２ ), -CH=CH-(CH ｐ ), and is ２ ), ｑ wherein X is NR ６ ,NHC(=O),C(=O)NH,O,SO ２ , a substituted or unsubstituted aryl ring, a substituted or unsubstituted heteroaryl ring, a substituted or unsubstituted heteroring, or a substituted or unsubstituted cyclic ring, Ring C is a cyclopropyl ring or a cyclobutyl ring. -C(R １ )- and -N(R ２ ) - dotted line connection 【Chemical 84】 Each of these is independently a single bond or a double bond, If the dotted line connection is a single connection, each R １ Each R is independently either H or OH, and each R ２ H is, If the dotted line connection is a double bond, each R １ H is H, and each R ２ It does not exist, R ３ and R ４ Each of these is independently H, NH ２ , NR ａ R ｂ OH, C １～４ Alkyl, C １～４ It is an alkoxy or aryl compound, R ａ and R ｂ These are H or C, respectively, independently. １～４ It is alkyl, R ５ H, C １～４ Alkyl, C １～４ It is an alkoxy or aryl compound, R ６ is H, or C １～４ It is alkyl, Each of m, n, and o is independently 1 or 2. Each of r, p, and q is an integer between 1 and 8, The compound, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, wherein the sum of p and q is an integer from 1 to 8.

2. Linker - (CH ２ ) ｒ The compound according to claim 1, wherein, in the case of -, ring A is of formula (IIa) and ring B is of formula (IIb).

3. Linker - (CH ２ ) ｐ -X-(CH ２ ) ｑ - or - (CH ２ ) ｐ -CH=CH-(CH ２ ) ｑ The compound according to claim 1, wherein, in the case of -, ring A is of formula (IIa), m in ring A is 2, and ring B is one of formulas (IIa), (IIb), (IIc), (IId), (IIe), (IIf), and (IIg).

4. Linker - (CH ２ ) ｐ -X-(CH ２ ) ｑ - or - (CH ２ ) ｐ -CH=CH-(CH ２ ) ｑ If - then ring A is of formula (IIa), m in ring A is 1, ring B is one of formulas (IIa), (IIb), (IIc), (IId), (IIe), (IIf), and (IIg), and formula (IIc) is either formula (IIc3) or formula (IIc4) below. 【Chemical 85】 The compound according to claim 1.

5. Xが、O、NR ６ 、NHC(=O)、-(m-C ６ H ４ )-、 【Chemical 86】 or 【Chemistry 87】 The compound according to any one of claims 1, 3, and 4.

6. Linker - (CH ２ ) ｒ - The compound according to claim 1.

7. The compound according to claim 1, 2, or 6, wherein r is 3 or 5.

8. The linker is - (CH ２ ) ｐ -O-(CH ２ ) ｑ - or - (CH ２ ) ｐ -NH-(CH ２ ) ｑ - The compound according to claim 1 or 5.

9. The compound according to any one of claims 1, 3 to 5, and 8, wherein the sum of p and q is 4.

10. Linker, 【Chemical 88】 The compound according to any one of claims 1, 3, and 4.

11. The compound according to claim 10, wherein the sum of p and q is 2.

12. Linker, 【Chemistry 89】 The compound according to claim 5.

13. The compound according to claim 12, wherein the sum of p and q is 2.

14. Linker - (CH ２ ) ｐ -CH=CH-(CH ２ ) ｑ - The compound according to claim 1, 3, or 4.

15. The compound according to claim 14, wherein the sum of p and q is 3.

16. The compound according to any one of claims 1 and 3 to 15, wherein ring B is of formula (IIa).

17. The compound according to claim 16, wherein m in ring B is 1.

18. The compound according to claim 16 or 17, wherein ring C in ring B is a cyclopropyl ring.

19. The dotted line connection within ring B is a single bond, R １ is H or OH, and R ２ The compound according to any one of claims 16 to 18, wherein is H.

20. The dotted bond within ring B is a double bond, R １ H is R ２ The compound according to any one of claims 16 to 18, wherein the compound is absent.

21. The compound according to claim 1, wherein ring A is of formula (IIb).

22. Ring A is of formula (IIb2): [Chemical 90] The compound according to claim 21.

23. R ３ The compound according to claim 21 or 22, wherein is H.

24. The dotted line connection within ring A is a single bond, R １ is H or OH, and R ２ The compound according to any one of claims 21 to 23, wherein is H.

25. The dotted bond within ring A is a double bond, R １ H is R ２ The compound according to any one of claims 21 to 23, wherein the compound is absent.

26. The aforementioned compound, 【Chemistry 91-1】 【Chemistry 91-2】 The compound according to claim 23, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof.

27. The compound according to claim 1, wherein ring A is of formula (IIg).

28. The compound according to claim 27, wherein the oxygen atoms in ring A are 2.

29. The dotted line connection within ring A is a single bond, R １ is H or OH, and R ２ The compound according to claim 27 or 28, wherein is H.

30. The dotted bond within ring A is a double bond, R １ H is R ２ The compound according to claim 27 or 28, wherein the compound is absent.

31. The aforementioned compound, 【Chemistry 92】 The compound according to claim 28, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof.

32. The compound according to claim 1, wherein ring A is of formula (IId).

33. The compound according to claim 32, wherein n in ring A is 1.

34. The dotted line connection within ring A is a single bond, R １ is H or OH, and R ２ The compound according to claim 32 or 33, wherein is H.

35. The dotted bond within ring A is a double bond, R １ H is R ２ The compound according to claim 32 or 33, wherein the compound is absent.

36. The aforementioned compound, 【Chemistry 93】 The compound according to claim 33, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof.

37. The compound according to claim 32, wherein n in ring A is 2.

38. The dotted line connection within ring A is a single bond, R １ is H or OH, and R ２ The compound according to claim 32 or 37, wherein is H.

39. The dotted bond within ring A is a double bond, R １ H is R ２ The compound according to claim 32 or 37, wherein the compound is absent.

40. The aforementioned compound, 【Chemical 94】 The compound according to claim 39, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof.

41. The compound according to claim 1, wherein ring A is of formula (IIc).

42. Ring A is of formula (IIc²): 【Chemical 95】 The compound according to claim 41.

43. R ４ is CH ３ O−, the compound according to claim 41 or 42.

44. The dotted line connection within ring A is a single bond, R １ is H or OH, and R ２ The compound according to any one of claims 41 to 43, wherein is H.

45. The dotted bond within ring A is a double bond, R １ H is R ２ The compound according to any one of claims 41 to 43, wherein the compound is absent.

46. The aforementioned compound, 【Chemistry 96】 The compound according to claim 42, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof.

47. The compound according to claim 1, wherein ring A is of formula (IIe).

48. R ５ The compound according to claim 47, wherein is methyl.

49. The dotted bond within ring A is a single bond, and R １ is H or OH, and R ２ is H, the compound according to claim 47 or 48.

50. The dotted bond within ring A is a double bond, R １ H is R ２ The compound according to claim 47 or 48, wherein the compound is absent.

51. The aforementioned compound, 【Chemistry 97】 The compound according to claim 48, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof.

52. The compound according to claim 1, wherein ring A and ring B are each independently of formula (IIa).

53. The compound according to claim 52, wherein m is 1.

54. The compound according to claim 52 or 53, wherein the ring C is a cyclopropyl ring.

55. The dotted line connection within ring A is a single bond, R １ is H or OH, and R ２ The compound according to any one of claims 52 to 54, wherein is H.

56. The dotted bond within ring A is a double bond, R １ H is R ２ The compound according to any one of claims 52 to 54, wherein the compound is absent.

57. The dotted line connection within ring B is a single bond, R １ is H or OH, and R ２ The compound according to any one of claims 52 to 56, wherein is H.

58. The dotted bond within ring B is a double bond, R １ H is R ２ The compound according to any one of claims 52 to 56, wherein the compound is absent.

59. The aforementioned compound, 【Chemistry 98-1】 【Chemistry 98-2】 The compound according to claim 54, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof.

60. The compound according to claim 1, wherein ring A and ring B are different.

61. Compounds of formula B(i) or B(ii): 【Chem.99】 or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, Each of rings A and B is independently one of the following equations: 【Chemistry 100-1】 【Chemistry 100-2】 【Chemistry 101】 This indicates the linking point to the linker or Ab linker. The linker is - (CH ２ ) ｒ -, - (CH ２ ) ｐ -X-(CH ２ ) ｑ - or - (CH ２ ) ｐ -CH=CH-(CH ２ ) ｑ - and X is NR ６ ,NHC(=O),C(=O)NH,O,SO ２ , a substituted or unsubstituted aryl ring, a substituted or unsubstituted heteroaryl ring, a substituted or unsubstituted heteroring, or a substituted or unsubstituted cyclic ring, Ring C is a cyclopropyl ring or a cyclobutyl ring. -C(R １ )- and -N(R ２ ) - dotted line connection 【Chemical Engineering 102】 Each of these is independently a single bond or a double bond, If the dotted line connection is a single connection, each R １ Each R is independently either H or OH, and each R ２ H is, If the dotted line connection is a double bond, each R １ H is independent of each R ２ It does not exist, R ３ and R ４ Each of these is independently H, NH ２ , NR ａ R ｂ OH, C １～４ Alkyl, C １～４ It is an alkoxyl or aryl, R ａ and R ｂ These are H or C, respectively, independently. １～４ It is alkyl, R ５ H, C １～４ Alkyl, C １～４ It is an alkoxyl or aryl, R ６ is H or C １～４ It is alkyl, Each of m, n, and o is independently 1 or 2. Each of r, p, and q is an integer between 1 and 8, The sum of p and q is an integer between 1 and 8. The Ab linker is a compound that can link ring A or ring B to a binder, the said compound, or a pharmaceutically acceptable salt thereof, tautomer, solvate, or stereoisomer.

62. The Ab linker has the following formula: 【Chemistry 103】 During the ceremony, 【Chemical 104】 The compound according to claim 61, wherein is a bonding site to ring A or ring B.

63. The aforementioned compound is given by the following formula: 【Chemistry 105】 or 【Chemistry 106】 The compound according to claim 61 or 62, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof.

64. Equation A(i) or A(ii) 【Chemistry 107】 A conjugate, or a pharmaceutically acceptable salt thereof, tautomer, solvate, or stereoisomer thereof, Each of rings A and B is independently one of the following equations: 【Chemistry 108-1】 【Chemistry 108-2】 【Chemistry 109】 This indicates the linking point to the linker or Ab linker. The linker is - (CH ２ ) ｒ -, - (CH ２ ) ｐ -X-(CH ２ ) ｑ - or - (CH ２ ) ｐ -CH=CH-(CH ２ ) ｑ - and X is NR ６ ,NHC(=O),C(=O)NH,O,SO ２ , a substituted or unsubstituted aryl ring, a substituted or unsubstituted heteroaryl ring, a substituted or unsubstituted heteroring, or a substituted or unsubstituted cyclic ring, Ring C is a cyclopropyl ring or a cyclobutyl ring. -C(R １ )- and -N(R ２ ) - dotted line connection 【Chemical 110】 Each of these is independently a single bond or a double bond, If the dotted line connection is a single connection, each R １ Each R is independently either H or OH, and each R ２ H is, If the dotted line connection is a double bond, each R １ H is independent of each R ２ It does not exist, R ３ and R ４ Each of these is independently H, NH ２ , NR ａ R ｂ OH, C １～４ Alkyl, C １～４ It is an alkoxyl or aryl, R ａ and R ｂ These are H or C, respectively, independently. １～４ It is alkyl, R ５ H, C １～４ Alkyl, C １～４ It is an alkoxyl or aryl, R ６ is H or C １～４ It is alkyl, Each of m, n, and o is independently 1 or 2. Each of r, p, and q is an integer between 1 and 8, The sum of p and q is an integer between 1 and 8. An Ab linker is a compound that links Ab to ring A or ring B. Ab is a binder selected from a humanized antibody, a chimeric antibody, a human antibody, or its antigen-binding fragment. The subscript x is 1 to 15, the conjugate, or a pharmaceutically acceptable salt thereof, tautomer, solvate, or stereoisomer.

65. The Ab linker has the following formula: 【Chemistry 111】 During the ceremony, 【Chemistry 112】 This indicates the connection point to Ab, 【Chemistry 113】 The conjugate according to claim 64, wherein indicates a bonding point to ring A or ring B.

66. The aforementioned conjugate is given by the following formula: 【Chemistry 114】 The conjugate according to claim 64 or 65, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, having the above.

67. The conjugate according to claim 66, wherein the subscript x is approximately 2.

68. The aforementioned conjugate is given by the following formula: 【Chemical 115】 A conjugate according to any one of claims 64 to 67, having the above, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof.

69. A pharmaceutical composition comprising a conjugate according to any one of claims 64 to 68, or a pharmaceutically acceptable salt thereof, tautomer, solvate, or stereoisomer, and a pharmaceutically acceptable excipient.

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