Anticancer nuclear hormone receptor targeting compounds

Abstract

Provided herein are compounds comprising a nuclear payload, such as a topoisomerase inhibitor, a topoisomerase toxic agent, or an analog thereof, and a nuclear receptor targeting epitope.

Description

[0001] Cross - reference to related applications

[0002] This application claims the priority and benefit of U.S. Provisional Patent Application No. 63 / 377,511, filed on September 28, 2022, the disclosure of which is incorporated herein by reference in its entirety. Background of the Invention

[0003] Topoisomerase inhibitors are compounds that block the action of topoisomerases, which are divided into two major subtypes: type I topoisomerase (TopI) and type II topoisomerase (TopII). Topoisomerases play important roles in cell replication and DNA organization because they mediate the cleavage of single - and double - stranded DNA to relax supercoils, unlink strands, and compact chromosomes in eukaryotic cells. Topoisomerase inhibitors affect these fundamental cellular processes. Some topoisomerase inhibitors prevent topoisomerases from making DNA strand breaks, while other topoisomerase inhibitors associate with topoisomerase - DNA complexes and prevent the religation step of the topoisomerase mechanism. These topoisomerase - DNA inhibitor complexes are cytotoxic agents because the unrepairable single - and double - stranded DNA breaks they cause can lead to apoptosis and cell death. Due to this apoptosis - inducing ability, topoisomerase inhibitors have attracted interest as therapies against infected and cancerous cells.

[0004] Camptothecin (CPT) is a topoisomerase poison. It is isolated from the bark and stems of Camptotheca acuminata (Camptotheca, Happy tree), a tree native to China, which is used in traditional Chinese medicine for cancer treatment. CPT has shown remarkable anti - cancer activity in preliminary clinical trials, especially against breast cancer, ovarian cancer, colon cancer, lung cancer, and gastric cancer. However, it has low solubility and has been reported to have adverse effects when used therapeutically. Therefore, synthetic and medicinal chemists have developed many syntheses of camptothecin and various derivatives to increase the benefits of this chemical and have achieved good results. Currently, four CPT analogs (topotecan, irinotecan, belotecan, and trastuzumab deruxtecan) have been approved and used in cancer chemotherapy. In addition to being an anti - tumor agent, camptothecin also shows anti - HIV activity because it interrupts the self - association of viral infectivity factors found in many retroviruses, including HIV.

[0005] There may also be many alternative uses for topoisomerase poisons in the future, including lupus, rare brain disorders, sepsis, and viral and trypanosomal infections. As additional roles of Top1 emerge, such as newly discovered regulatory functions, and Top1 continues to be associated with disease states, efforts will continue in the coming years for new drug discovery (and drug repurposing). Summary of the Invention

[0006] This disclosure provides compounds comprising a nuclear payload, such as a topoisomerase inhibitor, a topoisomerase poison, or an analogue thereof, and a nuclear receptor targeting epitope. The compounds described herein are designed to bind to nuclear receptors within a cell and allow the compound and its nuclear payload to accumulate in the cell nucleus. Without wishing to be bound by theory, one potential mode of enhanced utility is that this approach can provide compounds with cell type selectivity rather than just improved potency, thus working towards a higher therapeutic index. However, it is possible that the compounds can be active by other modes, such as but not limited to passive localization in the cell nucleus.

[0007] In addition, the compounds described herein provide targeted delivery of a nuclear payload. The compounds target and localize within tumor tissue. The compounds comprising at least one nuclear receptor targeting epitope (such as a nuclear steroid receptor targeting epitope) covalently attached to at least one nuclear payload are transported into the cell nucleus allowing the nuclear payload to accumulate in the cell nucleus, thereby enhancing tumor cell death. By doing so, the compounds described in this disclosure can exhibit excellent efficacy. In addition, the compounds described in this disclosure will spare cells that do not express a specific nuclear steroid receptor by accumulating in the cell nucleus of nuclear receptor positive cells (such as steroid receptor positive cells), and thus reduce side effects.

[0008] In certain embodiments, provided is a compound of Formula I, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue, or pharmaceutically acceptable salt thereof:

[0009] A 1 -L 1 -B 1 I

[0010] Wherein:

[0011] B 1 is a nuclear receptor targeting epitope;

[0012] L 1 is a covalent bond or a linking moiety; and

[0013] A 1 has Formula IA:

[0014]

[0015] Wherein:

[0016] R 1 、R 2 、R 3 、R 4 and R 5 are each independently hydrogen, a halogen group, a cyano group, a nitro group, -OR 15 、-SR 15 、-NR 15 R 16 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl, a 5- to 12-membered heteroaryl, -C(=O)R 15 、-C(=O)OR 15 、-OC(=O)R 15 、-OC(=O)NR 15 R 16 、-C(=O)NR 15 R 16 、-NR 15 C(=O)R 16 、-NR 15 C(=O)OR 16 、-S(=O) 1-2 R 15 、-S(=O) 1-2 NR 15 R 16 、-NR 15 S(=O) 1-2 R 16 、-Si(R 15 )3 or -C=NOR 15 , which are each independently optionally substituted by one or more R 10 when the valence allows;

[0017] Or R 1 and R 2 together with the atoms to which R 1 and R 2 are attached form a C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl or a 5- to 12-membered heteroaryl, which are each independently optionally substituted by one or more R 10 when the valence allows;

[0018] Or R 2and R 3 with R 2 and R 3 together with the attached atom form C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted by one or more R 10 substituents;

[0019] or R 3 and R 4 with R 3 and R 4 together with the attached atom form C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted by one or more R 10 substituents;

[0020] Each R 10 is independently a halogen group, cyano group, nitro group, -OR 17 , -SR 17 , -SF5, -NR 17 R 18 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, 5- to 12-membered heteroaryl, -C(=O)R 17 , -C(=O)OR 17 , -OC(=O)OR 17 , -OC(=O)R 17 , -C(=O)NR 17 R 18 , -OC(=O)NR 17 R 18 , -NR 7 C(=O)NR 17 R 18 , -S(=O) 1-2 R 17 , -S(=O) 1-2 NR 17 R 18 , -NR 17 S(=O) 1-2 R 18 , -NR 17 S(=O) 1-2 NR 17 R 18 , -NR 17 C(=O)R18 、-NR 17 C(=O)OR 18 、-Si(R 17 )3 or -C = NOR 17 , which are each independently optionally substituted by one or more substituents selected from the group consisting of halo, cyano, nitro, hydroxyl, amino, C 1-12 Alkoxy, C 1-12 Alkyl, C 2-12 Alkenyl, C 2-12 Alkynyl, C 3-12 Cycloalkyl, 5-12 membered heterocyclic group, C 6-12 Aryl and 5-12 membered heteroaryl;

[0021] Each R 15 and R 16 are independently hydrogen, C 1-12 Alkyl, C 2-12 Alkenyl, C 2-12 Alkynyl, C 3-12 Cycloalkyl, 5-12 membered heterocyclic group, C 6-12 Aryl or 5-12 membered heteroaryl, each independently optionally substituted by one or more substituents selected from the group consisting of halo, cyano, nitro, hydroxyl, amino, C 1-12 Alkoxy, C 1-12 Alkyl, C 2-12 Alkenyl, C 2-12 Alkynyl, C 3-12 Cycloalkyl, 5-12 membered heterocyclic group, C 6-12 Aryl and 5-12 membered heteroaryl;

[0022] or R 15 and R 16 With R 15 and R 16 The attached atoms together form a 5-12 membered heterocyclic group, which is optionally substituted, as valence permits, with one or more substituents selected from the group consisting of halo, cyano, nitro, hydroxy, amino, C 1-12 Alkoxy, C 1-12 Alkyl, C 2-12 Alkenyl, C 2-12 Alkynyl, C 3-12 Cycloalkyl, 5-12 membered heterocyclic group, C 6-12 Aryl and 5-12 membered heteroaryl; and

[0023] Each R 17 and R 18 are independently hydrogen, C 1-12 Alkyl, C 2-12 Alkenyl, C2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each independently optionally substituted, where valence allows, with one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl and 5- to 12-membered heteroaryl;

[0024] or R 17 and R 18 with R 17 and R 18 the atoms to which they are attached together form a 5- to 12-membered heterocyclic group, which is optionally substituted, where valence allows, with one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl and 5- to 12-membered heteroaryl;

[0025] where one or more atoms of formula IA (e.g., hydrogen, methyl or hydroxy) are replaced by a direct covalent bond to L 1 is provided.

[0026] There is also provided a compound of Table 1, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof.

[0027] There is also provided a pharmaceutical composition comprising a compound as described herein, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0028] There is also provided a method of treating or preventing cancer, the method comprising administering to an individual in need thereof an effective amount of a compound or composition as described herein. The cancer can be blood cancer, lung cancer, breast cancer, fallopian tube cancer, brain cancer, head and neck cancer, esophageal cancer, ovarian cancer, pancreatic cancer, peritoneal cancer, prostate cancer or skin cancer, such as but not limited to liver cancer, melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphocytic leukemia, chronic lymphocytic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, nephroblastoma, cervical cancer, testicular cancer, soft tissue sarcoma, chronic lymphocytic leukemia, Waldenström macroglobulinemia, primary macroglobulinemia, bladder cancer, chronic myelogenous leukemia, primary brain cancer, malignant melanoma, small cell lung cancer, gastric cancer, colon cancer, malignant pancreatic insulinoma, malignant carcinoid, malignant melanoma, choriocarcinoma, mycosis fungoides, head and neck cancer, osteogenic sarcoma, pancreatic cancer, acute granulocytic leukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma, genitourinary cancer, thyroid cancer, esophageal cancer, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, essential thrombocythemia, adrenocortical carcinoma, skin cancer, trophoblastic tumor or prostate cancer.

[0029] There is also provided a method of treating or preventing cancer, the method comprising administering to an individual in need thereof an effective amount of a compound or composition as described herein. Detailed Description

[0030] The following description sets forth exemplary embodiments of the technology of the present invention. However, it should be recognized that this description is not intended as a limitation on the scope of the present disclosure, but rather as a description of exemplary embodiments.

[0031] 1. Definitions

[0032] As used in this specification, the following words, phrases and symbols are generally intended to have the meanings set forth below, unless the context in which they are used indicates otherwise.

[0033] The term "about" refers to a variation of ±1%, ±3%, ±5% or ±10% of a specified value. For example, in some embodiments, "about 50" can include the range from 45 to 55. For integer ranges, the term "about" can include one or two integers greater than and / or less than the integers recited at the ends of the range. Unless otherwise indicated herein, the term "about" is intended to include values close to the recited ranges, such as weight percentages, which are equivalent in terms of the functionality of the individual components, compositions or embodiments. In addition, unless the context clearly dictates otherwise, the singular forms "a" and "the" include plural referents. Thus, for example, reference to "the compound" includes a plurality of such compounds, and reference to "the assay" includes reference to one or more compounds known to those skilled in the art and their equivalents.

[0034] "Alkyl" refers to an unbranched saturated hydrocarbon chain or a branched saturated hydrocarbon chain. As used herein, alkyl has from 1 to 12 carbon atoms ("C 1-12 alkyl"), from 1 to 10 carbon atoms (i.e., C 1-10 alkyl"), from 1 to 8 carbon atoms (i.e., C 1-8 alkyl"), from 1 to 6 carbon atoms (i.e., C 1-6 alkyl") or from 1 to 4 carbon atoms (i.e., C 1-4 alkyl"). Examples of alkyl include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl and 3-methylpentyl. When an alkyl residue having a specific number of carbon atoms is named by chemical name or identified by a molecular formula, all positional isomers having that number of carbon atoms can be encompassed; thus, for example, "butyl" includes n-butyl (i.e., -(CH2)3CH3), sec-butyl (i.e., -CH(CH3)CH2CH3), isobutyl (i.e., -CH2CH(CH3)2) and tert-butyl (i.e., -C(CH3)3); and "propyl" includes n-propyl (i.e., -(CH2)2CH3) and isopropyl (i.e., -CH(CH3)2).

[0035] "Haloalkyl" refers to an unbranched alkyl or branched alkyl as defined above, wherein one or more hydrogen atoms are replaced by a halogen. For example, when a residue is substituted by more than one halogen, it can be referred to by using a prefix corresponding to the number of attached halogen moieties. Dihaloalkyl and trihaloalkyl refer to an alkyl substituted by two ("di") or three ("tri") halo groups, which may or may not necessarily be the same halogen. Examples of haloalkyl include difluoromethyl (-CHF2) and trifluoromethyl (-CF3).

[0036] "Heteroalkyl" means an alkyl group in which one or more carbon atoms (and any associated hydrogen atoms) are each independently replaced by the same or different heteroatom groups. The term "heteroalkyl" includes straight-chain or branched-chain saturated chains having carbon and heteroatoms. For example, 1, 2, or 3 carbon atoms can be independently replaced by the same or different heteroatom groups. Heteroatom groups include, but are not limited to, -NH-, -O-, -S-, -S(O)-, -S(O)2-, etc. As used herein, heteroalkyl includes 1 to 8 carbon atoms, or 1 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom. "Heteroalkyl" means an alkyl group in which one or more carbon atoms (and any associated hydrogen atoms) are each independently replaced by the same or different heteroatom groups. The term "heteroalkyl" includes straight-chain or branched-chain saturated chains having carbon and heteroatoms. For example, 1, 2, or 3 carbon atoms can be independently replaced by the same or different heteroatom groups. Heteroatom groups include, but are not limited to, -NH-, -O-, -S-, -S(O)-, -S(O)2-. Examples of heteroalkyl include, for example, ethers (e.g., -CH2OCH3, -CH(CH3)OCH3, -CH2CH2OCH3, -CH2CH2OCH2CH2OCH3, etc.), thioethers (e.g., -CH2SCH3, -CH(CH3)SCH3, -CH2CH2SCH3, -CH2CH2SCH2CH2SCH3, etc.), sulfones (e.g., -CH2S(O)2CH3, -CH(CH3)S(O)2CH3, -CH2CH2S(O)2CH3, -CH2CH2S(O)2CH2CH2OCH3, etc.), and amines (e.g., -CH2NHCH3, -CH(CH3)NHCH3, -CH2CH2NHCH3, -CH2CH2NHCH2CH2NHCH3, etc.). As used herein, heteroalkyl includes 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom.

[0037] "Alkenyl" means an alkyl group containing at least one carbon-carbon double bond and having 2 to 20 carbon atoms (i.e., C 2-20 alkenyl), 2 to 8 carbon atoms (i.e., C 2-8 alkenyl), 2 to 6 carbon atoms (i.e., C 2-6 alkenyl), or 2 to 4 carbon atoms (i.e., C 2-4 alkenyl). Examples of alkenyl include, for example, vinyl, propenyl, and butadienyl (including 1,2-butadienyl and 1,3-butadienyl).

[0038] "Alkynyl" means an alkyl group containing at least one carbon-carbon triple bond and having 2 to 20 carbon atoms (i.e., C 2-20 alkynyl), 2 to 8 carbon atoms (i.e., C 2-8(alkynyl), 2 to 6 carbon atoms (i.e., C 2-6 (alkynyl) or 2 to 4 carbon atoms (i.e., C 2-4 (alkynyl) alkyl. The term "alkynyl" also includes those groups having one triple bond and one double bond.

[0039] "Alkoxy" refers to the group "alkyl-O-". Examples of alkoxy include, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentyloxy, n-hexyloxy, and 1,2-dimethylbutoxy.

[0040] "Alkoxyalkyl" refers to the group "alkyl-O-alkyl".

[0041] "Amino" refers to the group -NR y R z wherein R y and R z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroalkyl, or heteroaryl; as defined herein, each of which may optionally be substituted.

[0042] "Aryl" refers to an aromatic carbocyclic group having a single ring (e.g., monocyclic) or multiple rings (e.g., bicyclic or tricyclic), including fused systems. As used herein, aryl has 6 to 20 ring carbon atoms (i.e., C 6-20 aryl), 6 to 12 carbocyclic atoms (i.e., C 6-12 aryl), or 6 to 10 carbocyclic atoms (i.e., C 6-10 aryl). Examples of aryl include, for example, phenyl, naphthyl, fluorenyl, and anthracenyl. However, aryl does not encompass or overlap in any way with heteroaryl as defined below. If one or more aryl groups are fused to a heteroaryl group, the resulting ring system is heteroaryl. If one or more aryl groups are fused to a heterocyclic group, the resulting ring system is heterocyclic.

[0043] "Cycloalkyl" refers to a saturated or partially unsaturated cycloalkyl having a single ring or multiple rings (including fused, bridged, and spiro systems). The term "cycloalkyl" includes cycloalkenyl (i.e., a cyclic group having at least one double bond) and a carbocyclic fused ring system having at least one sp 3 carbon atom (i.e., at least one non-aromatic ring). As used herein, cycloalkyl has 3 to 20 ring carbon atoms (i.e., C 3-20 cycloalkyl), 3 to 12 ring carbon atoms (i.e., C 3-12 cycloalkyl), 3 to 10 ring carbon atoms (i.e., C 3-10 cycloalkyl), 3 to 8 ring carbon atoms (i.e., C 3-8 cycloalkyl), or 3 to 6 ring carbon atoms (i.e., C 3-6cycloalkyl). Monocyclic groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. In addition, the term cycloalkyl is intended to cover any non-aromatic ring that can be fused to an aryl ring, regardless of the attachment to the rest of the molecule. Further, when there are two substitution positions on the same carbon atom, cycloalkyl also includes "spirocycloalkyl".

[0044] "Heteroaryl" refers to an aromatic group having a single ring, multiple rings, or multiple fused rings, wherein one or more ring heteroatoms are independently selected from nitrogen, oxygen, and sulfur. As used herein, heteroaryl includes 1 to 20 ring carbon atoms (i.e., C 1-20 heteroaryl), 3 to 12 ring carbon atoms (i.e., C 3-12 heteroaryl), or 3 to 8 carbocyclic atoms (i.e., C 3-8 heteroaryl), and 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom, said ring heteroatoms being independently selected from nitrogen, oxygen, and sulfur. In certain cases, heteroaryl includes a 5-10 membered ring system, a 5-7 membered ring system, or a 5-6 membered ring system, each independently having 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom, said ring heteroatoms being independently selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl include, for example, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzonaphthofuranyl, benzoxazolyl, benzothienyl (benzothienyl) (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, isoquinolinyl, isoxazolyl, phthalazinyl, oxadiazolyl, oxazolyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, phenazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, thiazolyl, thiadiazolyl, thienyl (thienyl) (i.e., thiophenyl), triazolyl, tetrazolyl, and triazinyl. Examples of fused heteroaryl rings include, but are not limited to, benzo[d]thiazolyl, quinolinyl, isoquinolinyl, benzo[b]thienyl, indazolyl, benzo[d]imidazolyl, pyrazolo[1,5-a]pyridinyl, and imidazo[1,5-a]pyridinyl, wherein the heteroaryl can be attached through any ring of the fused system. Any aromatic ring having a single or multiple fused rings containing at least one heteroatom is considered a heteroaryl, regardless of the attachment to the rest of the molecule (i.e., through any one of the fused rings). Heteroaryl does not cover or overlap with aryl as defined above.

[0045] "Heterocyclic group" means a saturated or partially unsaturated cyclic alkyl group having one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur. The term "heterocyclic group" includes heterocyclic alkenyl (i.e., a heterocyclic group having at least one double bond), bridged heterocyclic group, fused heterocyclic group, and spiro heterocyclic group. The heterocyclic group can be a single ring or multiple rings, where the multiple rings can be fused, bridged, or spirocyclic, and can contain one or more (e.g., 1 to 3) oxo (=O) or N-oxide (-O - ) moieties. Any non-aromatic ring containing at least one heteroatom is considered a heterocyclic group, regardless of the attachment situation (i.e., it can be attached through a carbon atom or a heteroatom). In addition, the term heterocyclic group is intended to cover any non-aromatic ring containing at least one heteroatom, which can be fused to an aryl or heteroaryl ring, regardless of the attachment situation to the rest of the molecule. As used herein, the heterocyclic group has 2 to 20 ring carbon atoms (i.e., C 2-20 heterocyclic group), 2 to 12 ring carbon atoms (i.e., C 2-12 heterocyclic group), 2 to 10 ring carbon atoms (i.e., C 2-10 heterocyclic group), 2 to 8 ring carbon atoms (i.e., C 2-8 heterocyclic group), 3 to 12 ring carbon atoms (i.e., C 3-12 heterocyclic group), 3 to 8 ring carbon atoms (i.e., C 3-8 heterocyclic group), or 3 to 6 ring carbon atoms (i.e., C 3-6 heterocyclic group); having 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom, the ring heteroatoms being independently selected from nitrogen, sulfur, or oxygen. When there are two substitution positions on the same carbon atom, the term "heterocyclic group" also includes "spiro heterocyclic group". Examples of heterocyclic groups include, for example, azetidinyl, aza group, benzodioxolyl, benzo[b][1,4]dioxo Group, 1,4 - benzodioxanyl, benzopyranyl, benzodioxenyl, benzopyrone - yl, benzofuranone - yl, dioxolanyl, dihydropyranyl, hydropyranyl, thieno[1,3]dithiolanyl, decahydroisoquinolinyl, furanone - yl, imidazolinyl, imidazolidinyl, indolinyl, indazyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2 - oxopiperazinyl, 2 - oxopiperidinyl, 2 - oxopyrrolidinyl, oxazolidinyl, oxiranyl, oxetanyl, phenothiazinyl, phenoxazinyl, piperidinyl, piperazinyl, 4 - piperidinonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, trithiolanyl, tetrahydroquinolinyl, thiomorpholinyl, thiamorpholinyl, 1 - oxo - thiomorpholinyl and 1,1 - dioxo - thiomorpholinyl. When there are two substitution positions on the same carbon atom, the term "heterocyclic group" also includes "spiroheterocyclic group". Examples of spiro - heterocyclic rings include, for example, bicyclic and tricyclic systems such as oxabicyclo[2.2.2]octyl, 2 - oxa - 7 - azaspiro[3.5]nonyl, 2 - oxa - 6 - azaspiro[3.4]octyl and 6 - oxa - 1 - azaspiro[3.3]heptyl. Examples of fused heterocyclic rings include, but are not limited to, 1,2,3,4 - tetrahydroisoquinolinyl, 4,5,6,7 - tetrahydrothieno[2,3 - c]pyridinyl, indolinyl and isoindolinyl, where the heterocyclic group can be attached through any ring of the fused system.

[0046] "Alkylene" means a divalent alkyl group as defined above.

[0047] "Alkenylene" means a divalent alkenyl group as defined above.

[0048] "Alkynylene" means a divalent alkynyl group as defined above.

[0049] "Arylene" means a divalent aryl group as defined above.

[0050] "Cycloalkylene" means a divalent cycloalkyl group as defined above.

[0051] "Heterocycloalkylene" means a divalent heterocyclic group as defined above.

[0052] "Heteroarylene" means a divalent heteroaryl group as defined above.

[0053] "Oxo group" means = O.

[0054] "Halogen" or "halo group" includes fluorine, chlorine, bromine and iodine.

[0055] The term "optional" or "optionally" means that the subsequent described event or circumstance may or may not occur. The term "optionally substituted" means that any one or more hydrogen atoms on a specified atom or group may or may not be replaced by a moiety other than hydrogen.

[0056] As used herein, "substituted" means that one or more hydrogen atoms of a group are replaced by substituent atoms or groups commonly used in medicinal chemistry. Each substituent may be the same or different. Examples of suitable substituents include, but are not limited to, hydrazide, halo, -CN, -NO2, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclic, heteroaryl, -OR 56 , -C(O)OR 56 , -C(O)R 56 , -O-alkyl-OR 56 , -alkyl-OR 56 , haloalkyl, haloalkoxy, SR 56 , S(O)R 56 , SO2R 56 , NR 56 R 57 , -C(O)NR 56 R 57 , NR 56 C(O)R 57 , including its seleno and thio derivatives, wherein each R 56 and R 57 is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkyl-alkyl-, heterocyclic, heterocyclic-alkyl-, aryl, aryl-alkyl-, heteroaryl or heteroaryl-alkyl-, and wherein each substituent may optionally be further substituted.

[0057] Also provided are stereoisomers, mixtures of stereoisomers, tautomers, hydrates, solvates, isotopically enriched analogs and pharmaceutically acceptable salts of the compounds described herein.

[0058] The compounds or pharmaceutically acceptable salts thereof disclosed herein may include asymmetric centers and, thus, can give rise to enantiomers, diastereomers, and other stereoisomeric forms. In terms of absolute stereochemistry, the enantiomers, diastereomers, and other stereoisomeric forms may be defined as (R)- or (S)- or, for amino acids, as (D)- or (L)-. This disclosure is intended to embrace all such possible isomers, as well as their racemic and optically pure forms. The optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers can be prepared using chiral synthons or chiral reagents or resolved using conventional techniques (e.g., chromatography and fractional crystallization). Conventional techniques for the preparation / separation of individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high performance liquid chromatography (HPLC). When the compounds described herein contain an olefinic double bond or other geometrically asymmetric centers, the compounds are intended to include both E and Z geometric isomers unless otherwise specified.

[0059] "Stereoisomers" refer to compounds that are composed of the same atoms bonded by the same bonds but having different non-interchangeable three-dimensional structures. This disclosure contemplates various stereoisomers and mixtures thereof and includes "enantiomers" (which refer to two stereoisomers whose molecules are non-superimposable mirror images of each other) and "diastereomers" (which refer to stereoisomers that have at least two asymmetric atoms and are not mirror images of each other). Accordingly, all stereoisomers of the compounds of the present invention (e.g., geometric isomers, optical isomers, etc.) (including those of salts, solvates, and hydrates of the compounds) are contemplated, such as those that may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropisomers, and diastereomeric forms.

[0060] Diastereomer mixtures can be separated into their respective diastereomers based on their physicochemical differences by methods well known to those skilled in the art, such as, for example, by chromatography and / or fractional crystallization. Enantiomers are separated as follows: the enantiomer mixture is converted into a diastereomer mixture by reaction with a suitable optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), the diastereomers are separated, and the individual diastereomers are converted (e.g., hydrolyzed) into the corresponding pure enantiomers. In addition, some compounds may be atropisomers and are considered part of this disclosure. Stereoisomers can also be separated using chiral HPLC.

[0061] Some of the compounds exist as tautomers. Tautomers are in equilibrium with each other. For example, a compound containing an amide can exist in equilibrium with an imino acid tautomer. Regardless of which tautomer is shown and regardless of the nature of the equilibrium between the tautomers, one of ordinary skill in the art understands the compound to include both the amide and imino acid tautomers. Thus, a compound containing an amide is understood to include its imino acid tautomers. Similarly, a compound containing an imino acid is understood to include its amide tautomers.

[0062] The term “hydrate” refers to a complex formed by the combination of a compound and water as described herein.

[0063] “Solvate” refers to an association or complex of one or more solvent molecules with a compound of the present disclosure. Examples of solvents that form solvates include, but are not limited to, water, isopropyl alcohol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and ethanolamine.

[0064] Any compound or structure given herein is also intended to represent both the unlabeled form and the isotopically labeled form of the compound. These forms of the compound may also be referred to as “isotope-enriched analogs”. An isotopically labeled compound has the structure depicted herein, but one or more atoms are replaced with atoms having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I and 125 I. Various isotopically labeled compounds incorporated into the present disclosure, for example, incorporating radioactive isotopes such as 3 H and 14Those compounds of C. Such isotopically labeled compounds can be used in metabolic studies, reaction kinetics studies, detection or imaging techniques (such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including determination of drug or substrate tissue distribution), or in the radioactive treatment of patients. These compounds can exhibit increased metabolic resistance and can thus be used to increase the half-life of any compound when administered to mammals (especially humans). Such compounds are synthesized by means well known in the art, for example, by using starting materials in which one or more hydrogens have been replaced by deuterium.

[0065] Certain compounds disclosed herein contain one or more ionizable groups (groups from which a proton can be removed (e.g., -COOH) or added (e.g., amine) or that can be quaternized (e.g., amine)). All possible ionic forms of such molecules and their salts are intended to be included individually in the disclosure herein. Regarding the salts of the compounds described herein, one of ordinary skill in the art can select the appropriate ones from the various available counterions. In a particular application, the selection of a given anion or cation to prepare a salt may result in an increase or decrease in the solubility of the salt.

[0066] As used herein, the term "biologically non-cleavable linker moiety" is intended to mean a linker moiety that is not readily hydrolyzed under physiological conditions. As used herein, the term "biologically cleavable linker moiety" is intended to mean a linker moiety that is readily hydrolyzed under physiological conditions. In certain embodiments, at least one linker moiety is hydrolyzed under intracellular conditions (e.g., low pH). In some embodiments, the biological cleavage is self-cleavage and does not require physiological hydrolysis. In other embodiments, the cleavage of the biologically cleavable linker is initiated by metabolic activation, such as oxidative or pH-dependent cleavage without hydrolysis, e.g., by base- or acid-induced elimination, etc. More generally, a biologically cleavable linker can, in some cases, be analogous to a prodrug, where, upon cleavage, one or more drugs are released. In this sense, there are multiple mechanisms for cleaving a prodrug and releasing one or more active substances or precursors that produce active substances, and there are also multiple cleavable moieties known in the prodrug art and included in the definitions herein.

[0067] As used herein, the term "cancer" refers to a class of mammalian diseases characterized by uncontrolled cell growth. The term "cancer" may be used interchangeably with the terms "tumor", "solid tumor", "malignancy", "hyperproliferation", and "neoplasm". Cancer includes all types of hyperproliferative growth, proliferative growth, neoplastic growth, cancerous growth, or carcinogenic processes, metastatic tissues, or cells, tissues, or organs undergoing malignant transformation, regardless of histopathological type or stage of invasion. Exemplary examples include lung cancer, prostate cancer, head and neck cancer, breast cancer, and colorectal cancer, melanoma, and gliomas (such as high-grade gliomas, including glioblastoma multiforme (GBM), which is the most common and lethal malignant primary brain tumor in adults).

[0068] The phrase "solid tumor" includes, for example, lung cancer, head and neck cancer, brain cancer, oral cancer, colorectal cancer, breast cancer, prostate cancer, pancreatic cancer, and liver cancer. Other types of solid tumors are named after the specific cells that form them, for example, sarcomas formed by connective tissue cells (such as bone, cartilage, fat), carcinomas formed by epithelial tissue cells (such as breast, colon, pancreas), and lymphomas formed by lymphoid tissue cells (such as lymph nodes, spleen, and thymus). Regardless of the naming convention, the treatment of all types of solid tumors is within the scope of the present disclosure.

[0069] "Chemotherapeutic agent" refers to any substance capable of reducing or preventing the growth, proliferation, or spread of cancer cells, cancer cell populations, tumors, or other malignant tissues. The term is also intended to encompass radiotherapy or any anti-tumor or anti-cancer agent.

[0070] As used herein, "treatment" ("treatment" or "treating") is a method for obtaining a beneficial or desired result such as a clinical outcome. For the purposes of the present disclosure, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms and / or reduction in the degree of symptoms and / or prevention of worsening of symptoms associated with a disease or disorder. In one variant, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms and / or reduction in the degree of symptoms and / or prevention of worsening of symptoms associated with cognitive disorders, psychiatric disorders, neurotransmitter-mediated disorders, and / or neuronal disorders. In one embodiment, treating a disease or disorder with a compound of the present disclosure or a pharmaceutically acceptable salt thereof is not accompanied by side effects, or has fewer side effects than the currently available therapies for the disease or disorder, and / or improves the quality of life of an individual.

[0071] The terms “inhibit,” “inhibiting,” and “inhibition” refer to slowing, halting, or reversing the growth or progression of a disease, infection, disorder, or cell population. For example, inhibition can be greater than about 20%, 40%, 60%, 80%, 90%, 95%, or 99% compared to the growth or progression that occurs in the absence of treatment or exposure.

[0072] As used herein, “combination therapy” means a therapy that comprises two or more different compounds. Thus, in one aspect, there is provided a combination therapy that comprises a compound described herein and another compound. In some variations, the combination therapy optionally comprises one or more pharmaceutically acceptable carriers or excipients, non-pharmaceutically active compounds, and / or inert substances. In various embodiments, treatment with the combination therapy can result in additive or even synergistic (e.g., greater than additive) results compared to administration of a single compound of the present disclosure alone. In some embodiments, lower amounts of each compound are used as part of the combination therapy compared to the amounts typically used for monotherapy. In one embodiment, the same or greater therapeutic benefit is achieved using the combination therapy compared to using any single compound alone. In some embodiments, the same or greater therapeutic benefit is achieved using lower amounts of the compounds (e.g., lower doses or less frequent dosing schedules) in the combination therapy compared to the amounts typically used for the single compound or therapy. Preferably, the use of lower amounts of the compounds results in a reduction in the number, severity, frequency, and / or duration of one or more side effects associated with the compounds.

[0073] As used herein, the term “effective amount” refers to a parameter of the efficacy and toxicity of a compound of the present disclosure and such amount of the compound of the present disclosure that should be effective in a given therapeutic modality based on the knowledge of a practicing professional. As understood in the art, the effective amount can be one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired therapeutic endpoint. The effective amount can be considered in the context of administering one or more therapeutic agents, and a single agent can be considered to be administered in an effective amount if the desired or beneficial result can be achieved or realized in combination with one or more other agents. The appropriate dose of any co-administered compound can optionally be reduced due to the combined action of the compounds (e.g., additive or synergistic).

[0074] As used herein, IC 50 refers to the amount, concentration, or dose of a particular test compound that achieves 50% inhibition of the maximum response in an assay measuring a final response (e.g., modulation of PARP).

[0075] As used herein, EC 50Refers to the amount, concentration or dose of a particular test compound that elicits a dose-dependent response at 50% of the maximum expression of a particular response induced, stimulated or enhanced by the particular test compound.

[0076] As used herein, the term "cancer" refers to the abnormal growth of cells that tend to proliferate in an uncontrolled manner and, in some cases, metastasize (spread). Types of cancer include, but are not limited to, solid tumors (such as bladder, bowel, brain, breast, endometrium, heart, kidney, lung, lymphoid tissue (lymphoma), ovary, pancreas or other endocrine organs (thyroid)), prostate, skin (melanoma) or hematologic malignancies (such as leukemia).

[0077] As used herein, the term "carrier" refers to a relatively non-toxic compound or agent that facilitates the incorporation of a compound into cells or tissues.

[0078] As used herein, "unit dosage form" refers to physically discrete units suitable as unit doses, each unit containing a predetermined amount of an active ingredient calculated to produce the desired therapeutic effect in association with the desired pharmaceutical carrier. The unit dosage form may contain a single or combination therapy.

[0079] As used herein, the term "controlled release" refers to a drug-containing formulation or portion thereof in which the release of the drug is not immediate, i.e., administration via a "controlled release" formulation does not result in immediate release of the drug into the absorption pool. The term encompasses depot formulations designed to release the drug compound gradually over an extended period of time. Controlled release formulations can include a variety of drug delivery systems, typically involving mixing the drug compound with a carrier, polymer or other compound having the desired release characteristics (e.g., pH-dependent or non-pH-dependent dissolution, varying degrees of water solubility, etc.) and formulating the mixture according to the desired route of delivery (e.g., coated capsules, implantable reservoirs, injectable solutions containing biodegradable capsules, etc.).

[0080] As used herein, "pharmaceutically acceptable" or "pharmacologically acceptable" means a material that is not biologically or otherwise undesirable, e.g., the material can be incorporated into a pharmaceutical composition administered to a patient without causing any significant undesirable biological effects or interacting in a harmful manner with any other component of the composition containing the material. Pharmaceutically acceptable carriers or excipients preferably have met the required standards of toxicological and manufacturing testing, and / or are included in the Inactive Ingredient Guide compiled by the U.S. Food and Drug Administration.

[0081] "Pharmaceutically acceptable salts" are those salts that retain at least some of the biological activity of the free (non-salt) compound and can be administered to an individual as a drug (drug or pharmaceutical). Such salts include, for example: (1) acid addition salts formed from inorganic acids (such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.) or acid addition salts formed from organic acids (such as acetic acid, oxalic acid, propionic acid, succinic acid, maleic acid, tartaric acid, etc.); (2) salts formed when an acidic proton present in the parent compound is replaced by a metal ion (e.g., an alkali metal ion, an alkaline earth metal ion, or an aluminum ion) or salts formed when complexed with an organic base. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, etc. Acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, etc. Other examples of pharmaceutically acceptable salts include those listed in Berge et al., Pharmaceutical Salts, J. Pharm. Sci. 1977 Jan; 66(1):1-19. Pharmaceutically acceptable salts can be prepared in situ during the manufacturing process or by reacting the purified compound of the present disclosure in its free acid or base form with a suitable organic or inorganic base or organic or inorganic acid, respectively, and separating the salt thus formed during a subsequent purification process. It should be understood that reference to a pharmaceutically acceptable salt includes its solvated or crystalline forms, particularly solvates or polymorphs. Solvates contain a stoichiometric or non-stoichiometric amount of solvent and are typically formed during the crystallization process. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is alcohol. Polymorphs include different crystal packing arrangements of the same elemental composition of a compound. Polymorphs typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shapes, optical and electrical properties, stability, and solubility. Various factors (such as the recrystallization solvent, crystallization rate, and storage temperature) can all lead to the predominance of a single crystal form.

[0082] As used herein, the term "excipient" means an inert or inactive substance that can be used in the production of a drug (drug or pharmaceutical), such as a tablet containing a compound of the present disclosure as an active ingredient. The term excipient can encompass a variety of substances, including but not limited to any substance used as a binder, disintegrant, coating, compression / encapsulation aid, cream or emulsion, lubricant, solution for parenteral administration, material for chewable tablets, sweetening or flavoring agent, suspending / gelling agent or wet granulating agent. Binders include, for example, carbomer, povidone, xanthan gum, etc.; coatings include, for example, cellulose acetate phthalate, ethyl cellulose, gellan gum, maltodextrin, enteric coatings, etc.; compression / encapsulation aids include, for example, calcium carbonate, dextrose, fructose dc (directly compressible), honey dc, lactose (anhydrous or monohydrate; optionally in combination with aspartame, cellulose or microcrystalline cellulose), starch dc, sucrose, etc.; disintegrants include, for example, croscarmellose sodium, gellan gum, sodium carboxymethyl starch, etc.; creams or emulsions include, for example, maltodextrin, carrageenan, etc.; lubricants include, for example, magnesium stearate, stearic acid, sodium stearyl fumarate, etc.; materials for chewable tablets include, for example, dextrose, fructose dc, lactose (monohydrate, optionally in combination with aspartame or cellulose), etc.; suspending / gelling agents include, for example, carrageenan, sodium carboxymethyl starch, xanthan gum, etc.; sweetening agents include, for example, aspartame, dextrose, fructose dc, sorbitol, sucrose dc, etc.; and wet granulating agents include, for example, calcium carbonate, maltodextrin, microcrystalline cellulose, etc.

[0083] Compound

[0084] The present disclosure provides targeted compounds for treating cancer. The compounds described herein are capable of targeting the cell nucleus by recognition, and causing the nuclear receptor targeting epitope to bind to the corresponding binding site, and delivering a nuclear payload to the cell nucleus. The nuclear payload can then bind to one or more target sites within the cell nucleus and / or disrupt one or more cellular processes, thereby causing cell death.

[0085] In certain embodiments, the nuclear payload is bonded to one or more nuclear receptor targeting epitopes via a linking moiety. In certain embodiments, the linking moiety provides a single linkage or a single connection, meaning that the linker is conjugated to only one atom of each of the payload and the epitope.

[0086] Accordingly, there is provided a compound of formula I, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof:

[0087] A 1 -L 1 -B 1 I

[0088] Wherein:

[0089] B 1 is a nuclear receptor targeting epitope;

[0090] L 1 is a covalent bond or a linking moiety; and

[0091] A 1 has formula IA:

[0092]

[0093] Wherein:

[0094] R 1 , R 2 , R 3 and R 4 are each independently hydrogen, a halogen group, a cyano group, a nitro group, -OR 15 , -SR 15 , -NR 15 R 16 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl, a 5- to 12-membered heteroaryl, -C(=O)R 15 , -C(=O)OR 15 , -OC(=O)R 15 , -OC(=O)NR 15 R 16 , -C(=O)NR 15 R 16 , -NR 15 C(=O)R 16 , -NR 15 C(=O)OR 16 , -S(=O) 1-2 R 15 , -S(=O) 1-2 NR 15 R 16 , -NR 15 S(=O) 1-2 R 16 , -Si(R 15 )3 or -C=NOR 15 , which are each independently optionally substituted by one or more R 10 when the valence allows;

[0095] or R 1 and R 2 together with R1 and R 2 together with the attached atoms form a C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted by one or more R 10 substituents;

[0096] or R 2 and R 3 with R 2 and R 3 together with the attached atoms form a C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted by one or more R 10 substituents;

[0097] or R 3 and R 4 with R 3 and R 4 together with the attached atoms form a C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted by one or more R 10 substituents;

[0098] R 5 is hydrogen or -C(=O)R 15 ;

[0099] Each R 10 is independently a halogen group, cyano group, nitro group, -OR 17 , -SR 17 , -SF5, -NR 17 R 18 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, 5- to 12-membered heteroaryl, -C(=O)R 17 , -C(=O)OR 17 , -OC(=O)OR 17 , -OC(=O)R 17 , -C(=O)NR 17 R 18 , -OC(=O)NR 17 R 18 , -NR 7 C(=O)NR17 R 18 、 -S(=O) 1-2 R 17 、 -S(=O) 1-2 NR 17 R 18 、 -NR 17 S(=O) 1-2 R 18 、 -NR 17 S(=O) 1-2 NR 17 R 18 、 -NR 17 C(=O)R 18 、 -NR 17 C(=O)OR 18 、 -Si(R 17 )3 or -C=NOR 17 , which is independently optionally substituted, when the valence allows, by one or more substituents selected from the group consisting of: halo group, cyano group, nitro group, hydroxyl group, amino group, C 1-12 alkoxy group, C 1-12 alkyl group, C 2-12 alkenyl group, C 2-12 alkynyl group, C 3-12 cycloalkyl group, 5 - 12 - membered heterocyclic group, C 6-12 aryl group and 5 - 12 - membered heteroaryl group;

[0100] Each R 15 and R 16 is independently hydrogen, C 1-12 alkyl group, C 2-12 alkenyl group, C 2-12 alkynyl group, C 3-12 cycloalkyl group, 5 - 12 - membered heterocyclic group, C 6-12 aryl group or 5 - 12 - membered heteroaryl group, which is independently optionally substituted, when the valence allows, by one or more substituents selected from the group consisting of: halo group, cyano group, nitro group, hydroxyl group, amino group, C 1-12 alkoxy group, C 1-12 alkyl group, C 2-12 alkenyl group, C 2-12 alkynyl group, C 3-12 cycloalkyl group, 5 - 12 - membered heterocyclic group, C 6-12 aryl group and 5 - 12 - membered heteroaryl group;

[0101] or R 15 and R 16 together with R 15 and R 16The attached atoms together form a 5- to 12-membered heterocyclic group, which is optionally substituted, where valency permits, with one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl and 5- to 12-membered heteroaryl; and

[0102] each R 17 and R 18 is independently hydrogen, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, which is each independently optionally substituted, where valency permits, with one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl and 5- to 12-membered heteroaryl;

[0103] or R 17 and R 18 together with the atoms to which they are attached form a 5- to 12-membered heterocyclic group, which is optionally substituted, where valency permits, with one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 17 and R 18 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl and 5- to 12-membered heteroaryl;

[0104] wherein one or more atoms of formula IA (e.g., hydrogen, methyl or hydroxy) are replaced by a direct covalent bond to L 1 .

[0105] In certain embodiments, the compound is not a compound selected from the group of compounds in Table 1X, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof.​

[0106] Table 1 X

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113]

[0114] In certain embodiments, R 1 is hydrogen.

[0115] In certain embodiments, R 1 is C 1-12 alkyl, which is optionally substituted with one or more R 10 . In certain embodiments, R 1 is -Si(R 15 )3, which is optionally substituted with one or more R 10 .

[0116] In certain embodiments, R 1 is -C=NOR 15 , which is optionally substituted with one or more R 10 . In certain embodiments, R 1 is ethyl.

[0117] In certain embodiments, R 1 is In certain embodiments, R 1 is In certain embodiments, R 1 is In certain embodiments, R 1 is In certain embodiments, R 1 is

[0118] In certain embodiments, R 2 is hydrogen.

[0119] In certain embodiments, R 2 is C 1-12 alkyl, which is optionally substituted with one or more R 10 .

[0120] In certain embodiments, R 2 is

[0121] In certain embodiments, R 2 is nitro.

[0122] In certain embodiments, R 2 is

[0123] In certain embodiments, R 1 and R 2 together with the atoms to which R 1 and R 2 are attached form a C 3-12 cycloalkyl group, which is optionally substituted with one or more R 10 substituents.

[0124] In certain embodiments, R 1 and R 2 together with the atoms to which R 1 and R 2 are attached form

[0125] In certain embodiments, R 3 is -OR 15 , which is optionally substituted with one or more R 10 substituents.

[0126] In certain embodiments, R 3 is C 1-12 alkyl, which is optionally substituted with one or more R 10 substituents.

[0127] In certain embodiments, R 3 is -OC(=O)NR 15 R 16 , which is optionally substituted with one or more R 10 substituents.

[0128] In certain embodiments, R 3 is -OH.

[0129] In certain embodiments, R 3 is methyl.

[0130] In certain embodiments, R 3 is

[0131] In certain embodiments, R 3 is

[0132] In certain embodiments, R 3 is

[0133] In certain embodiments, R 3 is

[0134] In certain embodiments, R 3 is

[0135] In certain embodiments, R 3 is methoxy.

[0136] In certain embodiments, R 3 is hydrogen.

[0137] In certain embodiments, R 4 is hydrogen.

[0138] In certain embodiments, R 4 is a halo group.

[0139] In certain embodiments, R 3 and R 4 together with the atoms to which R 3 and R 4 are attached form a 5- to 12-membered heterocyclic group, which is optionally substituted with one or more R 10 substituents.

[0140] In certain embodiments, R 3 and R 4 together with the atoms to which R 3 and R 4 are attached form

[0141] In certain embodiments, R 5 is hydrogen.

[0142] In certain embodiments, R 5 is -C(=O)R 15 which is optionally substituted with one or more R 10 substituents.

[0143] In certain embodiments, R 5 is

[0144] In certain embodiments, R 5 is

[0145] In certain embodiments, A 1 is derived from:

[0146]

[0147]

[0148]

[0149] In certain embodiments, A 1 derived from:

[0150]

[0151]

[0152] In certain embodiments, the hydrogen atom of formula IA is replaced with a direct covalent bond to L 1 In certain embodiments, the methyl group of formula IA is replaced with a direct covalent bond to L 1 In certain embodiments, the hydroxyl group of formula IA is replaced with a direct covalent bond to L 1 In certain embodiments, L 1 is connected to the nitrogen atom of A 1 In certain embodiments, L 1 is connected to the oxygen atom of A 1

[0153] In certain embodiments, A 1 is:

[0154]

[0155]

[0156] In certain embodiments, A 1 is:

[0157]

[0158]

[0159] In certain embodiments, any compound disclosed herein (e.g., a compound of formula I) includes a topoisomerase inhibitor analog that, even after being modified to obtain the compounds described herein, still exhibits a biological activity comparable to the biological activity observed in the original, unmodified topoisomerase inhibitor. In certain embodiments, the topoisomerase inhibitor analog retains the ability to inhibit topoisomerase. In certain embodiments, the topoisomerase inhibitor analog exhibits a binding activity that is at least about 98%, about 95%, about 90%, about 85%, about 80%, about 75%, about 70%, about 65%, about 60%, about 55%, or about 50% of the binding activity observed in the original, unmodified topoisomerase inhibitor.​

[0160] Nuclear payload

[0161] In certain embodiments, the nuclear payload (i.e., A 1 ) in the compounds described herein is a topoisomerase inhibitor. As used herein, the term "topoisomerase inhibitor" refers to a chemical compound or moiety that blocks the action of a topoisomerase (or DNA topoisomerase), which is an enzyme involved in the overwinding or underwinding of DNA.

[0162] In certain embodiments, the nuclear payload (i.e., A 1 ) of the compounds described herein is derived from camptothecin (CPT). Thus, in certain embodiments, the nuclear payload (i.e., A 1 ) of the compounds described herein is a camptothecin (CPT) analogue. In certain embodiments, the nuclear payload (i.e., A 1 ) of the compounds described herein is derived from topotecan, irinotecan (CPT-11), silatecan (DB-67, AR-67), cositecan (BNP-1350), exatecan, lurtotecan, gimatecan (ST1481), belotecan (CKD-602) or rubitecan or an analogue thereof.

[0163] In certain embodiments, as used with respect to the nuclear payload (i.e., A 1 ) the term "derived from" or "analogue" means that at most one non-hydrogen atom of the original, unmodified nuclear payload (i.e., a known topoisomerase inhibitor) is replaced by a covalent bond (optionally via a linker moiety) to a nuclear receptor targeting epitope. Exemplary non-hydrogen atoms include, but are not limited to, -CH3, -OH, =O and -NH2. In certain embodiments, as used with respect to the nuclear payload (i.e., A 1 ) the term "derived from" means that one or more atoms (e.g., hydrogen, methyl or hydroxyl) of the original, unmodified nuclear payload (i.e., a topoisomerase inhibitor) are replaced by a direct covalent bond to L 1 . Exemplary non-hydrogen atoms include, but are not limited to, -CH3, -OCH3, -OH, =O, -NH2, -N(CH3)2, etc. In certain embodiments, one hydrogen atom of a heteroatom (e.g., N, O or S) that binds to the original, unmodified nuclear payload (i.e., a known topoisomerase inhibitor) is replaced by a covalent bond to L 1 . In certain embodiments, the term "derived from" means that one or more atoms (e.g., hydrogen, methyl or hydroxyl) are replaced by a covalent bond to L1 Direct covalent bond substitution.

[0164] In certain embodiments, one or more atoms (e.g., hydrogen, methyl, hydroxy, amino, etc.) on a nuclear payload as disclosed herein (i.e., A 1 ) are substituted to attach to the remainder of the compound (e.g., moiety -L 1 -B 1 ). In certain embodiments, a hydrogen atom on a nuclear receptor targeting epitope as disclosed herein is substituted to attach to the remainder of the compound. In certain embodiments, the hydrogen atom is on a heteroatom. In certain embodiments, the hydrogen atom is on a halogen. In certain embodiments, the hydrogen atom is on nitrogen. In certain embodiments, the hydrogen atom is on oxygen. In certain embodiments, the hydrogen atom is on carbon (e.g., methyl). The analogs are derived from known nuclear payloads as described herein (e.g., topoisomerase inhibitors or A 1 ) and are modified to optionally conjugate to at least one nuclear hormone receptor targeting epitope via a linking moiety. The analogs retain a biological activity comparable to that observed in the original, unmodified topoisomerase inhibitor even after being modified to obtain the compounds described herein. In certain embodiments, the compounds exhibit a binding activity or inhibition that is at least about 98%, about 95%, about 90%, about 85%, about 80%, about 75%, about 70%, about 65%, about 60%, about 55%, or about 50% or about 5%-50% of the binding activity or inhibition observed in the original, unmodified topoisomerase inhibitor. In certain embodiments, the compounds as described herein exhibit an IC 50 of less than about 500 nM, or less than about 400 nM, or less than about 350 nM, or less than about 300 nM, or less than about 200 nM, or less than about 100 nM, or less than about 50 nM.

[0165] In certain embodiments, the nuclear payload (i.e., A 1 ) is derived from:

[0166]

[0167]

[0168] In certain embodiments, the nuclear payload or A 1 is derived from:

[0169]

[0170]

[0171] Nuclear hormone receptor targeting epitope

[0172] In certain embodiments, B 1 is a nuclear hormone receptor targeting epitope. In certain embodiments, B 1 binds to an estrogen receptor, a glucocorticoid receptor, a progesterone receptor, or an androgen receptor. In certain embodiments, B 1 binds to an estrogen receptor. In certain embodiments, B 1 binds to a glucocorticoid receptor. In certain embodiments, B 1 binds to a progesterone receptor. In certain embodiments, B 1 binds to an androgen receptor. Exemplary estrogen receptor, glucocorticoid receptor, progesterone receptor, or androgen receptor binders are described herein.

[0173] In certain embodiments, B 1 is a nuclear steroid receptor targeting epitope. As used herein, "nuclear receptor targeting epitope" refers to the following portion of the compounds described herein (e.g., B 1 ), which is derived from a nuclear targeting agent as disclosed herein and interacts with the ligand-binding domain of a target nuclear receptor, i.e., the portion of the compound that drives the ligand-binding interaction. Nuclear receptor targeting epitopes are used to associate compounds with target nuclear receptors (e.g., nuclear steroid receptors), facilitate the localization of compounds to nuclear steroid receptor-expressing cells, and translocate nuclear payloads from the cytoplasm to the nucleus, thereby accumulating the compounds in the nucleus. The level of accumulation can be controlled by selecting an appropriate nuclear receptor targeting epitope. For example, through the nuclear translocation of nuclear steroid receptors that occurs after the epitope binds to the receptor, the compounds described herein can accumulate in the nucleus to different extents, with a high level of accumulation in the case of a full agonist (e.g., dihydrotestosterone (DHT)), a medium level of accumulation in the case of a partial agonist (e.g., bicalutamide), and a low level of accumulation in the case of an antagonist (e.g., enzalutamide).

[0174] Steroid receptor targets can be any steroid receptor, including but not limited to those overexpressed in cancer cells. In certain embodiments, at least one nuclear steroid receptor targeting epitope is capable of binding to the ligand-binding domain of a nuclear steroid receptor, such as the ligand-binding domain on an estrogen receptor, a glucocorticoid receptor, a progesterone receptor, or an androgen receptor.

[0175] Exemplary nuclear steroid receptor - targeting epitopes include those derived from: androgen receptor agonists, androgen receptor antagonists, selective androgen receptor modulators (SARMs), estrogen receptor agonists, estrogen receptor antagonists, selective estrogen receptor modulators (SERMs), glucocorticoid receptor antagonists, glucocorticoid receptor agonists, selective glucocorticoid receptor modulators (SGRMs), progesterone receptor antagonists, progesterone receptor agonists, selective progesterone receptor modulators (SPRMs), or combinations thereof.

[0176] Nuclear steroid receptor - targeting epitopes are generally capable of binding to nuclear steroid receptors, with an IC 50 less than about 500 nM, or less than about 400 nM, or less than about 300 nM, or less than about 200 nM, or less than about 100 nM, or an EC 50 less than about 1 μM, or less than about 900 nM, or less than about 800 nM, or less than about 700 nM, or less than about 600 nM, or less than about 500 nM, or less than about 400 nM, or less than about 3400 nM, or less than about 200 nM, or less than about 100 nM.

[0177] In certain embodiments, the nuclear hormone receptor - binding affinity of the compounds of the invention can be defined in terms of their affinity relative to a reference nuclear hormone receptor - binding compound. For example, some compounds of the invention can bind to the estrogen receptor. In some cases, the compounds disclosed herein bind to the human estrogen receptor with an affinity of at least 0.1%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the affinity of 17β - estradiol.

[0178] As another example, some compounds of the invention can bind to the human androgen receptor. In some cases, the compounds disclosed herein bind to the androgen receptor with an affinity of at least 0.1%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the affinity of dihydrotestosterone (DHT).

[0179] As another example, some compounds of the invention can bind to the human progesterone receptor. In some cases, the compounds disclosed herein bind to the progesterone receptor with an affinity of at least 0.1%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the affinity of progesterone.

[0180] Alternatively, for example, some compounds of the invention can bind to the human glucocorticoid receptor. In some cases, the compounds disclosed herein bind to the glucocorticoid receptor with an affinity of at least 0.1%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% of the affinity of cortisol.

[0181] In certain embodiments, the nuclear steroid receptor targeting epitope (e.g., B 1 ) is an agonist of the androgen receptor. In certain embodiments, the nuclear steroid receptor targeting epitope is an antagonist of the androgen receptor.

[0182] In certain embodiments, the nuclear steroid receptor targeting epitope (e.g., B 1 ) is steroidal (or derived from a steroidal compound) (e.g., dihydrotestosterone). In certain embodiments, the nuclear steroid receptor targeting epitope is non-steroidal (or derived from a non-steroidal compound) (e.g., enzalutamide, apalutamide, AZD9496, and bicalutamide).

[0183] Analogs are derived from known nuclear steroid receptor targeting epitopes described herein (e.g., B 1 ) and are modified to optionally conjugate to at least one nuclear steroid payload via a linking moiety. The analogs retain a biological activity comparable to the biological activity observed in the original, unmodified nuclear steroid receptor targeting epitope even after being modified to obtain the compounds described herein. In certain embodiments, the compounds exhibit a binding activity or inhibition that is at least about 98%, about 95%, about 90%, about 85%, about 80%, about 75%, about 70%, about 65%, about 60%, about 55% or about 50% or about 5%-50% of the binding activity or inhibition observed in the original, unmodified nuclear steroid receptor targeting epitope.

[0184] In certain embodiments, the analogs are derived from known nuclear receptor targeting epitopes (e.g., B 1 ), such as known nuclear steroid receptor targeting epitopes. In certain embodiments, B 1Bind to estrogen receptors, glucocorticoid receptors, progesterone receptors, or androgen receptors. In certain embodiments, as used with respect to a nuclear receptor targeting epitope, the term "derived from" means that at most one non-hydrogen atom of an original, unmodified nuclear receptor targeting compound (i.e., a known nuclear steroid receptor targeting compound) is replaced with a covalent bond to a nuclear payload (optionally via a linker moiety). Exemplary non-hydrogen atoms include, but are not limited to, -CH3, -OH, =O, and -NH2. In certain embodiments, as used with respect to a nuclear receptor targeting epitope, the term "derived from" means that at most one non-hydrogen atom of an original, unmodified nuclear receptor targeting compound (i.e., a known nuclear steroid receptor targeting compound) is replaced with a covalent bond to a nuclear payload (optionally via a linker moiety). In certain embodiments, a hydrogen atom bonded to a heteroatom (e.g., N, O, or S) of an original, unmodified nuclear receptor targeting compound (i.e., a known nuclear steroid receptor targeting compound) is replaced with a covalent bond to a nuclear payload (optionally via a linker moiety). In certain embodiments, the term "derived from" means that one or more atoms (e.g., hydrogen, methyl, or hydroxy) are replaced with a direct covalent bond to L 1 is replaced with a direct covalent bond.

[0185] In certain embodiments, the nuclear steroid receptor targeting epitope (e.g., B 1 )) is an androgen receptor targeting epitope. As used herein, the term "androgen receptor targeting epitope" is intended to mean the portion of a compound that binds to the androgen receptor and can be functionally an androgen receptor agonist or an androgen receptor antagonist (including a partial androgen receptor agonist or a partial androgen receptor antagonist), and in some embodiments, is capable of binding to the receptor and shuttling the ligand-receptor complex from the cytoplasm to the cell nucleus. "Androgen receptor" (AR) - also known as NR3C4 (nuclear receptor subfamily 3C member 4) is a type of nuclear receptor that can translocate androgens to the nucleus when activated by binding an androgen receptor binder (e.g., an androgen such as testosterone or dihydrotestosterone) in the cytoplasm.

[0186] In certain embodiments, a single atom on the nuclear receptor targeting epitope (B 1 ) as disclosed herein is replaced to attach to the remainder of the compound (e.g., moiety -L 1 -B 1 ). In certain embodiments, a halogen atom on the nuclear receptor targeting epitope as disclosed herein is replaced to attach to the remainder of the compound. In certain embodiments, a hydrogen atom on the nuclear receptor targeting epitope as disclosed herein is replaced to attach to the remainder of the compound. In certain embodiments, the hydrogen atom is on a heteroatom. In certain embodiments, the hydrogen atom is on nitrogen. In certain embodiments, the hydrogen atom is on oxygen. In certain embodiments, the hydrogen atom is on carbon.

[0187] In certain embodiments, B 1 has Formula IIA:

[0188]

[0189] wherein:

[0190] The wavy bond represents the point of attachment to L 1 ;

[0191] R 30 is hydrogen, C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, C 2-12 alkynyl, or C 3-12 cycloalkyl, wherein when valence allows, each C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, C 2-12 alkynyl, or C 3-12 cycloalkyl is optionally independently substituted with one or more R 100 ;

[0192] R 40 is hydrogen, C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, C 2-12 alkynyl, or C 3-12 cycloalkyl, wherein when valence allows, each C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, C 2-12 alkynyl, or C 3-12 cycloalkyl is optionally independently substituted with one or more R 100 ;

[0193] Each R 50 and R 51 is independently a halogen, cyano, nitro, -OR 170 , -SR 170 , -NR 170 R 180 , C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, or C 2-12 alkynyl; wherein when valence allows, each C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, or C 2-12The alkynyl group is independently optionally substituted by one or more halo groups, hydroxy groups or amino groups;

[0194] Each R 100 is independently an oxo group, a halo group, a cyano group, a nitro group, -OR 170 , -SR 170 , -SF5, -NR 170 R 180 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl, a 5- to 12-membered heteroaryl, -C(=O)R 170 , -C(=O)OR 170 , -OC(=O)OR 170 , -OC(=O)R 170 , -C(=O)NR 170 R 180 , -OC(=O)NR 170 R 180 , -NR 170 C(=O)NR 170 R 180 , -S(=O) 1-2 R 170 , -S(=O) 1-2 NR 170 R 180 , -NR 170 S(=O) 1-2 R 180 , -NR 170 S(=O) 1-2 NR 170 R 180 , -NR 170 C(=O)R 180 or -NR 170 C(=O)OR 180 , which are each independently optionally substituted by one or more substituents selected from the group consisting of: halo group, cyano group, nitro group, hydroxy group, amino group, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl and a 5- to 12-membered heteroaryl; and

[0195] Each R 170 and R 180 is independently hydrogen or C 1-12An alkyl group which is optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group when the valence allows.

[0196] Or R 170 And R 180 With R 170 And R 180 The attached atoms together form a heterocyclic group optionally substituted by a halogen group or a C 1-12 alkyl group optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group.

[0197] In certain embodiments, B 1 is

[0198] In certain embodiments, B 1 has the formula IIB' or IIB":

[0199]

[0200] Wherein:

[0201] The wavy bond represents the point of attachment to L 1 ;

[0202] R N is H or C 1-12 alkyl;

[0203] R 60 is hydrogen, -OR 101 、-NR 101 R 102 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5-12 membered heterocyclic group, C 6-12 aryl, 5-12 membered heteroaryl, -C(=O)R 101 、-C(=O)OR 101 、-OC(=O)R 101 、-OC(=O)NR 101 R 102 、-C(=O)NR 101 R 102 、-NR 101 C(=O)R 102 、-NR 101 C(=O)OR 102 , each of which is optionally independently substituted by one or more R 100 ;

[0204] R 80 is hydrogen, -OR 101 、-NR101 R 102 、 C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, 5- to 12-membered heteroaryl, -C(=O)R 101 、 -C(=O)OR 101 、 -OC(=O)R 101 、 -OC(=O)NR 101 R 102 、 -C(=O)NR 101 R 102 、 -NR 101 C(=O)R 102 、 -NR 101 C(=O)OR 102 , which are each optionally and independently substituted by one or more R 100 substituents, where valence allows;

[0205] R 81 is hydrogen, -OR 101 、 -NR 101 R 102 、 C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, 5- to 12-membered heteroaryl, -C(=O)R 101 、 -C(=O)OR 101 、 -OC(=O)R 101 、 -OC(=O)NR 101 R 102 、 -C(=O)NR 101 R 102 、 -NR 101 C(=O)R 102 、 -NR 101 C(=O)OR 102 , which are each optionally and independently substituted by one or more R 100 substituents, where valence allows;

[0206] or R 80 and R 81 attached to the atom to which R 80 and R 81 are attached together form a heterocyclic group optionally substituted by a halogen group or a C 1-12 alkyl group optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group;

[0207] R 82 is hydrogen, -OR 101 , -NR 101 R 102 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5 - 12 - membered heterocyclic group, C 6-12 aryl, 5 - 12 - membered heteroaryl, -C(=O)R 101 , -C(=O)OR 101 , -OC(=O)R 101 , -OC(=O)NR 101 R 102 , -C(=O)NR 101 R 102 , -NR 101 C(=O)R 102 , -NR 101 C(=O)OR 102 , which are each independently optionally substituted by one or more R 100 substituents when the valence allows;

[0208] Each R 101 and R 102 is independently hydrogen, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5 - 12 - membered heterocyclic group, C 6-12 aryl or 5 - 12 - membered heteroaryl, which are each independently optionally substituted by one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5 - 12 - membered heterocyclic group, C 6-12 aryl and 5 - 12 - membered heteroaryl;

[0209] Each R 100 is independently oxo, halo, cyano, nitro, -OR 170 , -SR 170 , -SF5, -NR 170 R 180 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5 - 12 - membered heterocyclic group, C 6-12Aryl, 5-12 membered heteroaryl, -C(=O)R 170 、-C(=O)OR 170 、-OC(=O)OR 170 、-OC(=O)R 170 、-C(=O)NR 170 R 180 、-OC(=O)NR 170 R 180 、-NR 170 C(=O)NR 170 R 180 、-S(=O) 1-2 R 170 、-S(=O) 1-2 NR 170 R 180 、-NR 170 S(=O) 1-2 R 180 、-NR 170 S(=O) 1-2 NR 170 R 180 、-NR 170 C(=O)R 180 or -NR 170 C(=O)OR 180 , which are each independently optionally substituted by one or more substituents selected from the group consisting of halo, cyano, nitro, hydroxyl, amino, C 1-12 Alkoxy, C 1-12 Alkyl, C 2-12 Alkenyl, C 2-12 Alkynyl, C 3-12 Cycloalkyl, 5-12 membered heterocyclic group, C 6-12 Aryl and 5-12 membered heteroaryl; and

[0210] Each R 170 and R 180 are independently hydrogen or C 1-12 Alkyl, which is optionally substituted with oxo, halo, hydroxyl or amino where valency permits,

[0211] or R 170 and R 180 With R 170 and R 180 The attached atoms together form a heterocyclyl optionally substituted by halo or a C optionally substituted by oxo, halo, hydroxy or amino. 1-12 alkyl.

[0212] In certain embodiments, B 1 It has the formula IIB'.

[0213] In certain embodiments, B 1 has formula IIB”.

[0214] In certain embodiments, R N is methyl.

[0215] In certain embodiments, B 1 has formula IIB:

[0216]

[0217] wherein:

[0218] The wavy bond represents the point of attachment to L 1 ;

[0219] R 60 is hydrogen, -OR 101 , -NR 101 R 102 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5-12 membered heterocyclic group, C 6-12 aryl, 5-12 membered heteroaryl, -C(=O)R 101 , -C(=O)OR 101 , -OC(=O)R 101 , -OC(=O)NR 101 R 102 , -C(=O)NR 101 R 102 , -NR 101 C(=O)R 102 , -NR 101 C(=O)OR 102 , each of which is optionally independently substituted with one or more R 100 ;

[0220] R 80 is hydrogen, -OR 101 , -NR 101 R 102 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5-12 membered heterocyclic group, C 6-12 aryl, 5-12 membered heteroaryl, -C(=O)R 101 , -C(=O)OR 101 , -OC(=O)R 101 , -OC(=O)NR 101R 102 、 -C(=O)NR 101 R 102 、 -NR 101 C(=O)R 102 、 -NR 101 C(=O)OR 102 , which is each optionally and independently substituted by one or more R 100 substituents;

[0221] R 81 is hydrogen, -OR 101 、 -NR 101 R 102 、 C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5 - 12 - membered heterocyclic group, C 6-12 aryl, 5 - 12 - membered heteroaryl, -C(=O)R 101 、 -C(=O)OR 101 、 -OC(=O)R 101 、 -OC(=O)NR 101 R 102 、 -C(=O)NR 101 R 102 、 -NR 101 C(=O)R 102 、 -NR 101 C(=O)OR 102 , which is each optionally and independently substituted by one or more R 100 substituents;

[0222] or R 80 and R 81 attached to R 80 and R 81 together with the atoms to which they are attached form a heterocyclic group optionally substituted by a halogen group or a C 1-12 alkyl optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group;

[0223] R 82 is hydrogen, -OR 101 、 -NR 101 R 102 、 C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5 - 12 - membered heterocyclic group, C 6-12 aryl, 5 - 12 - membered heteroaryl, -C(=O)R 101 、 -C(=O)OR 101, -OC(=O)R 101 , -OC(=O)NR 101 R 102 , -C(=O)NR 101 R 102 , -NR 101 C(=O)R 102 , -NR 101 C(=O)OR 102 , which are each independently optionally substituted by one or more R 100 substituents, where valence allows;

[0224] Each R 101 and R 102 is independently hydrogen, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, or 5- to 12-membered heteroaryl, which are each independently optionally substituted by one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, and 5- to 12-membered heteroaryl;

[0225] Each R 100 is independently oxo, halo, cyano, nitro, -OR 170 , -SR 170 , -SF5, -NR 170 R 180 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, 5- to 12-membered heteroaryl, -C(=O)R 170 , -C(=O)OR 170 , -OC(=O)OR 170 , -OC(=O)R 170 , -C(=O)NR 170 R 180 , -OC(=O)NR 170 R 180 , -NR 170 C(=O)NR 170 R 180, -S(=O) 1-2 R 170 , -S(=O) 1-2 NR 170 R 180 , -NR 170 S(=O) 1-2 R 180 , -NR 170 S(=O) 1-2 NR 170 R 180 , -NR 170 C(=O)R 180 or -NR 170 C(=O)OR 180 , which are each independently optionally substituted, when valency permits, with one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocycloalkyl, C 6-12 aryl and 5- to 12-membered heteroaryl; and

[0226] each R 170 and R 180 is independently hydrogen or C 1-12 alkyl, which is optionally substituted, when valency permits, with oxo, halo, hydroxy or amino,

[0227] or R 170 and R 180 together with the atoms to which they are attached form a heterocycloalkyl optionally substituted with halo or a C 170 alkyl optionally substituted, when valency permits, with oxo, halo, hydroxy or amino. 180 1-12

[0228] In certain embodiments, R 82 is hydrogen. In certain embodiments, R 82 is C 1-12 alkyl. In certain embodiments, R 82 is methyl.

[0229] In certain embodiments, B 1 is

[0230] In certain embodiments, B 1 is

[0231] In certain embodiments, B​​1 is

[0232] In certain embodiments, B 1 is

[0233] In certain embodiments, B 1 is

[0234] In certain embodiments, B 1 is

[0235] In certain embodiments, B 1 has the formula IIC':

[0236]

[0237] wherein:

[0238] The wavy bond represents the point of attachment to L 1 ;

[0239] A” and A”' are each independently O or S;

[0240] R a and R b are each independently CH3 or CH2CH3; or R a and R b together with the atoms to which R a and R b are attached form a C 3-6 cycloalkyl, oxirane, oxetane or tetrahydrofuran;

[0241] B, B 10 , B 2 , B 3 , B', B 1’ , B 2’ and B 3’ are each independently CR c or N;

[0242] Each R c is independently hydrogen, fluorine, CN or methyl;

[0243] D is absent, or is NH, O, S, CH2, -NH(C=O)-, -(C=O)NH- or C=O;

[0244] X” is CN, halo or NO2;

[0245] Y” is CH3, CH2R d , CHF2 or CF3;

[0246] R d is a halogenated group;

[0247] "Z” is H, C 1-2 alkyl, C2 alkenyl or NO2; or

[0248] X” and Y” together form where the dash indicates a bond to the ring;

[0249] or Y” and Z” together form where each is a single bond or a double bond, and where the dash indicates a bond to the ring; and

[0250] Z' is CH or N.

[0251] In certain embodiments, D is NH, O, S, CH2, -NH(C=O)-, -(C=O)NH- or C=O.

[0252] In certain embodiments, B 1 has formula IIC:

[0253]

[0254] where:

[0255] The wavy bond represents the point of attachment to L 1 ;

[0256] A” and A”' are each independently O or S;

[0257] R a and R b are each independently CH3 or CH2CH3; or R a and R b together with the atoms to which R a and R b are attached form a C 3-6 cycloalkyl, oxirane, oxetane or tetrahydrofuran;

[0258] B, B 10 、B 2 、B 3 、B', B 1’ 、B 2’ and B 3’ are each independently CR c or N;

[0259] Each R c is independently hydrogen, fluorine, CN or methyl;

[0260] D is NH, O, S, CH2, -NH(C=O)-, -(C=O)NH- or C=O;

[0261] "X" is CN, a halogenated group, or NO2;

[0262] "Y" is CH3, CH2R d , CHF2, or CF3;

[0263] R d is a halogenated group;

[0264] "Z" is H, C 1-2 alkyl, C2 alkenyl, or NO2; or

[0265] "X" and "Y" together form where the dash indicates the bond to the ring;

[0266] or "Y" and "Z" together form where each is a single bond or a double bond, and where the dash indicates the bond to the ring; and

[0267] Z' is CH or N.

[0268] In certain embodiments, B 1 is

[0269] In certain embodiments, B 1 is

[0270] In certain embodiments, B 1 is

[0271] In certain embodiments, B 1 has the formula IID':

[0272]

[0273] where:

[0274] W is O, S, or NH;

[0275] Each is independently a double bond or a single bond;

[0276] Each R 61 and R 62 is independently hydrogen, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, or C 3-12 cycloalkyl, where when valence allows, each C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, or C 3-12 cycloalkyl is optionally independently substituted with one or more R100 Substitute;

[0277] Each R 100 is independently an oxo group, a halo group, a cyano group, a nitro group, -OR 170 , -SR 170 , -SF5, -NR 170 R 180 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl, a 5- to 12-membered heteroaryl, -C(=O)R 170 , -C(=O)OR 170 , -OC(=O)OR 170 , -OC(=O)R 170 , -C(=O)NR 170 R 180 , -OC(=O)NR 170 R 180 , -NR 170 C(=O)NR 170 R 180 , -S(=O) 1-2 R 170 , -S(=O) 1-2 NR 170 R 180 , -NR 170 S(=O) 1-2 R 180 , -NR 170 S(=O) 1-2 NR 170 R 180 , -NR 170 C(=O)R 180 or -NR 170 C(=O)OR 180 , which are each independently optionally substituted, when valency permits, by one or more substituents selected from the group consisting of: halo group, cyano group, nitro group, hydroxy group, amino group, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl and a 5- to 12-membered heteroaryl; and

[0278] Each R 170 and R 180 is independently hydrogen or C 1-12An alkyl group which is optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group when valence allows.

[0279] Or R 170 And R 180 Attached to R 170 And R 180 The atoms together form a heterocyclic group optionally substituted by a halogen group or a C 1-12 alkyl group optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group.

[0280] In certain embodiments, B 1 Has the formula IID:

[0281]

[0282] Wherein:

[0283] W is O, S or NH;

[0284] Is a double bond or a single bond;

[0285] Each R 61 And R 62 Independently is hydrogen, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl or C 3-12 cycloalkyl, wherein when valence allows, each C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl or C 3-12 cycloalkyl is optionally independently substituted by one or more R 100 substituents;

[0286] Each R 100 Independently is an oxo group, a halogen group, a cyano group, a nitro group, -OR 170 、-SR 170 、-SF5、-NR 170 R 180 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl, a 5- to 12-membered heteroaryl, -C(=O)R 170 、-C(=O)OR 170 、-OC(=O)OR 170 、-OC(=O)R 170 、-C(=O)NR 170 R 180 、-OC(=O)NR170 R 180 、-NR 170 C(=O)NR 170 R 180 、-S(=O) 1-2 R 170 、-S(=O) 1-2 NR 170 R 180 、-NR 170 S(=O) 1-2 R 180 、-NR 170 S(=O) 1-2 NR 170 R 180 、-NR 170 C(=O)R 180 or -NR 170 C(=O)OR 180 , which are each independently optionally substituted by one or more substituents selected from the group consisting of halo, cyano, nitro, hydroxyl, amino, C 1-12 Alkoxy, C 1-12 Alkyl, C 2-12 Alkenyl, C 2-12 Alkynyl, C 3-12 Cycloalkyl, 5-12 membered heterocyclic group, C 6-12 Aryl and 5-12 membered heteroaryl;

[0287] Each R 170 and R 180 are independently hydrogen or C 1-12 Alkyl, which is optionally substituted with oxo, halo, hydroxyl or amino where valency permits,

[0288] or R 170 and R 180 With R 170 and R 180 The attached atoms together form a heterocyclyl optionally substituted by halo or a C optionally substituted by oxo, halo, hydroxy or amino. 1-12 alkyl.

[0289] In certain embodiments, B 1 yes

[0290] In certain embodiments, B 1 yes

[0291] In certain embodiments, B 1 With formula IIE:

[0292]

[0293] Wherein:

[0294] The tilde key refers to the connection point with L;

[0295] Q is wherein key a is attached to ring a and key b is attached to ring b;

[0296] R a and R b are each independently -CH3 or -CH2CH3; or R a and R b together with the atom to which R a and R b is attached form a C 3-5 cycloalkyl, oxiranyl, oxetanyl or tetrahydrofuranyl;

[0297] A and A' are each independently O or S;

[0298] E, E 1 、E 2 and E 3 are each independently CR c or N, and each R c is independently hydrogen, halo, CN or methyl;

[0299] E 4 is CF, CH or N;

[0300] Q 1 is a bond, CH2, C=O or (C=O)NH;

[0301] Q 2 is NH, O, S, CH2, NH(C=O), C(=O)NH or C=O;

[0302] R 44 、R 45 and R 46 are each independently hydrogen, CN or C 1-2 alkyl;

[0303] t is 0, 1, 2, 3 or 4;

[0304] Each R e and R f is independently halo, cyano, C 1-4 alkyl or C 1-4 haloalkyl;

[0305] R 41 is halo, CN or NO2;

[0306] R 42 is a halogen group, CH3, CH2F, CHF2 or CF3; or

[0307] R 41 and R 42 together form wherein the dash indicates the bond to ring a;

[0308] R 43 is hydrogen, a halogen group, C 1-2 alkyl, C2 alkenyl, NO2, CF3; or

[0309] R 42 and R 43 together form wherein each is a single bond or a double bond, and wherein the dash indicates the bond to ring a.

[0310] In certain embodiments, B 1 is

[0311] In certain embodiments, B 1 is

[0312] In certain embodiments, B 1 is

[0313] In certain embodiments, B 1 is

[0314] In certain embodiments, B 1Derived from progesterone, enobosarm, bicalutamide, apalutamide, testosterone, dihydrotestosterone, testosterone, 19-nortestosterone, progesterone, andarine, cortisol, prednisone, flutamide, nilutamide, enzalutamide, tamoxifen, toremifene, raloxifene, bazedoxifene, ospemifene, megestrol acetate, estramustine, abiraterone, LGD-2941, BMS-564929, ostarine, ulipristal acetate, asoprisnil (J867), mifepristone, telapristone (CDB-4124, Proellex, Progenta) or analogs thereof.

[0315] In certain embodiments, B 1 comprises a nuclear receptor targeting epitope derived from:

[0316]

[0317]

[0318]

[0319] or a stereoisomer or mixture of stereoisomers or an analog thereof, wherein at least one hydrogen atom is replaced with a direct covalent bond to A 1 (optionally via a linking moiety).

[0320] These and other Bs that can be described herein 1Selective androgen receptor modulators (SARMs) used as nuclear steroid receptor targeting epitopes can be found in: US 6,462,038, US 6,777,427, WO2001 / 027086, WO2004 / 013104, WO2004 / 000816, WO2004 / 0113309, US2006 / 0211756, US2006 / 0063819, US2005 / 245485, US2005 / 250741, US2005 / 277681, WO2006 / 060108, WO2004 / 041277, WO2003 / 034987, US2006 / 0148893, US2006 / 0142387, WO2005 / 000795, WO2005 / 085185, WO2006 / 133216, WO2006 / 044707, WO2006 / 124447, WO2007 / 002181, WO2005 / 108351, WO2005 / 115361, and US2006 / 0160845.

[0321] In certain embodiments, B 1 is a selective estrogen receptor modulator (SERM). In certain embodiments, B 1Epitopes derived from the following: anordrin, bazedoxifene, broparestrol (Acnestrol), clomifene (Clomid), cyclofenil (Sexovid), lasofoxifene (Fablyn), ormeloxifene (Centron, Novex, Novex-DS, Sevista), ospemifene (Osphena, deaminohydroxytoremifene), raloxifene (Evista), tamoxifen (Nolvadex), toremifene (Fareston; 4-chlorotamoxifen), acolbifene, afimoxifene (4-hydroxytamoxifen; metabolite of tamoxifen), elacestrant, enclomifene ((E)-clomifene), endoxifen (4-hydroxy-N-demethyltamoxifen; metabolite of tamoxifen), zuclomifene ((Z)-clomifene), bazedoxifene, arzoxifene, brilanestrant, clomifenoxide (clomifene N-oxide; metabolite of clomifene), droloxifene (3-hydroxytamoxifen), etacstil, fispemifene, GW-7604 (4-hydroxyetacstil), idoxifene (pyrrolidino-4-iodotamoxifen), levormeloxifene ((L)-ormeloxifene), miproxifene, nafoxidine, nitromifene (CI-628), panomifene, pipendoxifene (ERA-923), trioxifene, keoxifene, LY117018, onapristone, fareston (toremifene citrate) or zindoxifene (D-16726) or analogs thereof.

[0322] In certain embodiments, the SERM is structurally classified as a triphenylethylene (tamoxifen, clomiphene, toremifene, droloxifene, idoxifene, ospemifene, nafoxidine, arzoxifene, etc. or analogs thereof), a benzothiophene (raloxifene, arzoxifene, etc. or analogs thereof), an indole (bazedoxifene, indoxifene, pipendoxifene, etc. or analogs thereof), a tetrahydronaphthalene (lasofoxifene, nafoxidine, etc. or analogs thereof), or a benzopyran (acobifene, ormeloxifene, levormeloxifene, etc. or analogs thereof).

[0323] In certain embodiments, B 1 is a selective estrogen receptor downregulator (SERD). In certain embodiments, the compound comprises at least one nuclear steroid receptor targeting epitope, which independently comprises an epitope derived from: fulvestrant, blestran (ARN-810), itesetide (GW5638), AZD9496, giredestrant (GDC-9545), or GW7604.

[0324] In certain embodiments, B 1 is a selective progesterone receptor modulator (SPRM). In certain embodiments, B comprises an epitope derived from: ulipristal acetate, asoprisnil (J867), mifepristone, trimegestone (CDB-4124, Proellex, Progenta), or analogs thereof.

[0325] In certain embodiments, B 1 comprises an epitope derived from: estrogen, estetrol, estriol, estrone, progesterone, embosamol, bicalutamide, apalutamide, testosterone, dihydrotestosterone, estradiol, flutamide, nilutamide, enzalutamide, tamoxifen, toremifene, raloxifene, bazedoxifene, ospemifene, megestrol acetate, estramustine, abiraterone, LGD-2941, BMS-564929, ostarine, or analogs thereof.

[0326] In certain embodiments, at least one nuclear steroid receptor targeting epitope is an androgen receptor targeting epitope and comprises:

[0327]

[0328]

[0329] or a stereoisomer or a mixture of stereoisomers or an analog thereof, wherein the wavy line indicates the point of attachment to the nuclear payload (optionally via a linking moiety).

[0330] In certain embodiments, at least one nuclear steroid receptor targeting epitope is an estrogen receptor targeting epitope and comprises:

[0331]

[0332]

[0333] or a stereoisomer or mixture of stereoisomers or an analogue thereof, wherein the wavy line indicates the point of attachment to the nuclear payload (optionally via a linker moiety).

[0334] In certain embodiments, at least one nuclear steroid receptor targeting epitope is an estrogen receptor targeting epitope and comprises:

[0335]

[0336]

[0337] or a stereoisomer or mixture of stereoisomers or an analogue thereof, wherein the wavy line indicates the point of attachment to the nuclear payload (optionally via a linker moiety).

[0338] In certain embodiments, at least one nuclear steroid receptor targeting epitope comprises:

[0339]

[0340]

[0341]

[0342] or a stereoisomer or mixture of stereoisomers or an analogue thereof, wherein the wavy line indicates the point of attachment to the nuclear payload (optionally via a linker moiety).

[0343] In certain embodiments, at least one nuclear steroid receptor targeting epitope comprises:

[0344]

[0345]

[0346] or a stereoisomer or mixture of stereoisomers or an analogue thereof, wherein the wavy line indicates the point of attachment to the nuclear payload (optionally via a linker moiety).

[0347] In certain embodiments, the nuclear steroid receptor targeting epitope is not or does not contain a peptide, protein, nanoparticle or antibody.

[0348] Linker moiety

[0349] The "linking moiety" of any compound described herein can be biocleavable (e.g., acid-labile) or non-biocleavable. The linking moiety can be linear, branched, saturated, unsaturated, all-carbon or heteroatom-containing. The linking moiety can also contain one or more fused, saturated, unsaturated, and all-carbon or heteroatom-containing rings. In certain embodiments, the linking moiety is a non-biocleavable linking moiety. In certain embodiments, the linking moiety is a biocleavable linking moiety. In certain embodiments, the core payload is bonded to one nuclear steroid receptor targeting epitope via a non-biocleavable linking moiety and to one or more nuclear steroid receptor targeting epitopes via a biocleavable linking moiety. In certain embodiments, the biocleavable linking moiety is an acid-labile linking moiety. In some embodiments, the linking moiety includes a hydrazone linkage.

[0350] It is contemplated that any linking moiety can be used in the compounds described herein provided that it does not significantly interfere with or disrupt the desired binding of the core payload or the nuclear receptor targeting epitope.

[0351] In certain embodiments, L 1 has the formula:

[0352] -(L a ) q -,

[0353] wherein:

[0354] each L a is independently W, -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -NR 110 S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -C(O)O-, C 1-12 alkylene, C 2-12 alkenylene, C 2-12 alkynylene, C 6-12 arylene, C 3-12A subcycloalkyl group, a 5-12 membered hetero-subcycloalkyl group or a 5-12 membered hetero-aryl group, each independently optionally substituted with one or more substituents independently selected from: -OH, -NH2, -CN, oxo group, halogen group, C 1-4 alkyl group, C 1-4 haloalkyl group, C 1-4 alkoxy group, C 1-4 haloalkoxy group, C 6-12 aryl group, 5-12 membered hetero-aryl group, C 3-12 cycloalkyl group and 5-12 membered heterocyclic group,

[0355] wherein each W is independently wherein R n is independently H, C 1-4 alkyl group or C 1-4 haloalkyl group; and wherein R w is independently H, C 3-12 cycloalkyl group, C optionally substituted with one or more halogen groups or OH 6-12 aryl group or C optionally substituted with one or more substituents independently selected from: 1-4 alkyl group: halogen group, OH, -SH, -S(C 1-4 alkyl group), -CONH2, -COOH, -NHC(=NH)NH2, -NH2, -NHCOCH3, -NHCHO, -NHCONH2, C 6-12 aryl group, 5-12 membered heterocyclic group or 5-12 membered hetero-aryl group;

[0356] each R 110 is independently hydrogen, C 1-4 alkyl group, C 1-4 haloalkyl group, C 1-4 alkoxy group, C 1-4 haloalkoxy group, C 6-12 aryl group, 5-12 membered hetero-aryl group, C 3-12 cycloalkyl group or 5-12 membered heterocyclic group;

[0357] each R 120 is independently hydrogen, C 1-4 alkyl group, C 1-4 haloalkyl group, C 1-4 alkoxy group, C 1-4 haloalkoxy group, C 6-12 aryl group, 5-12 membered hetero-aryl group, C 3-12 cycloalkyl group or 5-12 membered heterocyclic group; and

[0358] q is an integer from 0 to 40. It should be understood that either end of L 1 can be connected to A 1 .

[0359] In certain embodiments, L 1 has the formula:

[0360] -(L a ) q -,

[0361] wherein:

[0362] each L a is independently -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -NR 110 S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -C(O)O-, C 1-12 alkylene, C 2-12 alkenylene, C 2-12 alkynylene, C 6-12 arylene, C 3-12 cycloalkylene, 5- to 12-membered hetero cycloalkylene or 5- to 12-membered heteroarylene, each of which is independently optionally substituted with one or more substituents independently selected from: oxo group, halo group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocyclic group;

[0363] each R 110 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocyclic group;

[0364] each R 120 is independently hydrogen, C1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; and

[0365] q is an integer from 0 to 20.

[0366] In certain embodiments, L 1 has the formula:

[0367] -Y 10 -(CHR 130 ) n’ -Y 20 -(CHR 140 ) n” -Y 30 -(CHR 150 ) m” -Y 40 -(CHR 160 ) p -Y 50 -

[0368] (CHR 170 ) p' -Y 60 -

[0369] wherein:

[0370] each Y 10 , Y 20 , Y 30 , Y 40 , Y 50 and Y 60 is independently -(W) s -, a bond, -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -NR 110 S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120-, -C(O)-, -OC(O)-, -OC(O)O-, -(CH2CH2O) 1-5 -, -C(O)O-, C 1-12 alkylene, C 2-12 alkenylene, C 2-12 alkynylene, C 6-12 arylene, C 3-12 cycloalkylene, 5- to 12-membered hetero cycloalkylene or 5- to 12-membered heteroarylene, each independently optionally substituted with one or more substituents independently selected from: -OH, -NH2, -CN, oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocycloalkyl;

[0371] Each W is independently wherein R n is independently H, C 1-4 alkyl or C 1-4 haloalkyl; and wherein R w is independently H, C 3-12 cycloalkyl, C optionally substituted with one or more halo or OH 6-12 aryl or C optionally substituted with one or more substituents independently selected from: 1-4 alkyl: halo, OH, -SH, -S(C 1-4 alkyl), -CONH2, -COOH, -NHC(=NH)NH2, -NH2, -NHCOCH3, -NHCHO, -NHCONH2, C 6-12 aryl, 5- to 12-membered heterocycloalkyl or 5- to 12-membered heteroaryl;

[0372] Each R 110 , R 120 , R 130 , R 140 , R 150 , R 160 and R 170 are independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12A cycloalkyl group or a 5-12 membered heterocyclic group, each independently optionally substituted with one or more substituents independently selected from the following: -OH, -NH2, -CN, oxo group, halogen group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5-12 membered heteroaryl, C 3-12 cycloalkyl and 5-12 membered heterocyclic group; and

[0373] n', n", m", s, p and p' are each independently 0, 1, 2, 3, 4, 5, 6, 7 or 8.

[0374] In certain embodiments, L 1 has the formula:

[0375] -Y 10 -(CHR 130 ) n '-Y 20 -(CHR 140 ) n” -Y 30 -(CHR 150 ) m” -Y 40 -

[0376] Wherein:

[0377] Each Y 10 , Y 20 , Y 30 and Y 40 is independently a bond, -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -NR 110 S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -(CH2CH2O) 1-5 -, -C(O)O-, C 1-12 alkylene, C 2-12 alkenylene, C 2-12Alkynylene, C 6-12 Arylene, C 3-12 Cycloalkylene, 5- to 12-membered hetero cycloalkylene or 5- to 12-membered heteroarylene, each independently optionally substituted with one or more substituents independently selected from: oxo, halo, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy or C 1-4 Haloalkoxy;

[0378] Each R 110 Independently is hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, C 6-12 Aryl, 5- to 12-membered heteroaryl, C 3-12 Cycloalkyl or 5- to 12-membered heterocycloalkyl;

[0379] Each R 120 Independently is hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, C 6-12 Aryl, 5- to 12-membered heteroaryl, C 3-12 Cycloalkyl or 5- to 12-membered heterocycloalkyl;

[0380] Each R 130 Independently is hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, C 6-12 Aryl, 5- to 12-membered heteroaryl, C 3-12 Cycloalkyl or 5- to 12-membered heterocycloalkyl;

[0381] Each R 140 Independently is hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, C 6-12 Aryl, 5- to 12-membered heteroaryl, C 3-12 Cycloalkyl or 5- to 12-membered heterocycloalkyl;

[0382] Each R 150 Independently is hydrogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, C6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; and

[0383] n', n” and m” are each independently 0, 1, 2, 3, 4, 5, 6, 7 or 8.

[0384] In certain embodiments, at least one W is Val. In certain embodiments, at least one W is Cit. In certain embodiments, s is 2. In certain embodiments, -(W) s - is -Val-Cit-.

[0385] In certain embodiments, L 1 has the formula:

[0386]

[0387] wherein:

[0388] each L 2 、L 3 and L 4 is independently a bond, C 1-12 alkylene, -NHC(=O)-, -C(=O)NH-, -C(=O)-O-, -O-C(=O)- or C=O;

[0389] each R 200 and R 201 is independently a halogen group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocycloalkyl; and

[0390] each s and s' is independently 0, 1, 2, 3 or 4.

[0391] In certain embodiments, L 1 has the formula:

[0392] -L 2 -L 3 -Cy1-L 4 -Cy2-L 5 -L 6 -

[0393] wherein:

[0394] each L 2 、L 3 、L 4 、L5 and L 6 is independently a bond, C 1-12 alkylene, -O-, -NHC(=O)-, -C(=O)NH-, -C(=O)-O-, -O-C(=O)- or C=O, wherein one or more carbon atoms in the C 1-12 alkylene are optionally replaced by oxygen;

[0395] Cy1 and Cy2 are each independently a bond, C 6-12 arylene, C 3-12 cycloalkylene, 5- to 12-membered hetero-cycloalkylene or 5- to 12-membered heteroarylene, wherein each group is independently optionally substituted by one or more substituents independently selected from the following: -OH, -NH2, -CN, oxo group, halogen group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocyclic group.

[0396] In certain embodiments, Cy1 is a 5- to 12-membered hetero-cycloalkylene optionally substituted by one or more substituents independently selected from the following: -OH, -NH2, -CN, oxo group, halogen group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocyclic group.

[0397] In certain embodiments, Cy1 is optionally substituted by one or more substituents independently selected from the following -OH, -NH2, -CN, oxo group, halogen group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocyclic group. In certain embodiments, Cy1 is optionally substituted by one or more substituents independently selected from the following -OH, -NH2, -CN, oxo group, halogen group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl, and 5- to 12-membered heterocycloalkyl. In certain embodiments, Cy1 is In certain embodiments, Cy1 is In certain embodiments, Cy1 is a bond. In certain embodiments, Cy1 is or a bond.

[0398] In certain embodiments, Cy2 is a 5- to 12-membered heterosubcycloalkyl optionally substituted with one or more substituents independently selected from: -OH, -NH2, -CN, oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl, and 5- to 12-membered heterocycloalkyl.

[0399] In certain embodiments, Cy2 is optionally substituted with one or more substituents independently selected from -OH, -NH2, -CN, oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl, and 5- to 12-membered heterocycloalkyl. In certain embodiments, Cy2 is optionally substituted with one or more substituents independently selected from -OH, -NH2, -CN, oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl, and 5- to 12-membered heterocycloalkyl. In certain embodiments, Cy2 is In certain embodiments, Cy2 is In certain embodiments, Cy2 is a bond. In certain embodiments, Cy2 is or a bond.

[0400] In certain embodiments, the linking moiety has the formula:

[0401]

[0402] Wherein:

[0403] Ring C is a 3- to 12-membered cycloalkylene or 3- to 12-membered hetero-cycloalkylene, each independently optionally substituted with one or more substituents independently selected from: oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy or C 1-4 haloalkoxy;

[0404] Each Y 50 and Y 60 is independently a bond, -NR 110 , -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -NR 110 S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -(CH2CH2O) 1-5 (-), -C(O)O-, C 1-12 alkylene, C 2-12 alkenylene, C 2-12 alkynylene, C 6-12 arylene, C 3-12 cycloalkylene, 5- to 12-membered hetero-cycloalkylene or 5- to 12-membered hetero-arylene, each independently optionally substituted with one or more substituents independently selected from: oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy or C 1-4 haloalkoxy;

[0405] Each R 110 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocyclo;

[0406] Each R 120Independently is hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5 - 12 - membered heteroaryl, C 3-12 cycloalkyl or 5 - 12 - membered heterocycloalkyl; and

[0407] wherein “*” and the wavy line represent covalent bonds.

[0408] In certain embodiments, each C of Y 50 and Y 60 alkylidene, C 1-12 alkenylidene, C 2-12 alkynylidene, C 2-12 arylidene, C 6-12 cycloalkylidene, 5 - 12 - membered hetero - cycloalkylidene or 5 - 12 - membered heteroarylidene is independently optionally substituted with one to five substituents independently selected from: halo, C 3-12 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy or C 1-4 haloalkoxy. 1-4

[0409] In certain embodiments, the linking moiety has the formula:

[0410]

[0411] wherein:

[0412] Ring C is a 3 - 12 - membered cycloalkylidene or 3 - 12 - membered hetero - cycloalkylidene, each of which is independently optionally substituted with one or more substituents independently selected from: oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy or C 1-4 haloalkoxy;

[0413] Each Y 50 and Y 60 is independently a bond, - NR 110 -, - O -, - S(O) 0-2 -, - NR 110 C(O) -, - C(O)NR 110 -, - NR 110 C(O)NR 110 -, - NR 110 S(O)2 -, - S(O)2NR 110 -, - NR 110 S(O)2NR 110 ​-, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -(CH2CH2O) 1-5 -, -C(O)O-, C 1-12 alkylene, C 2-12 alkenylene, C 2-12 alkynylene, C 6-12 arylene, C 3-12 cycloalkylene, 5- to 12-membered hetero-substituted cycloalkylene or 5- to 12-membered heteroarylene, each independently optionally substituted with one or more substituents independently selected from: oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy or C 1-4 haloalkoxy;

[0414] Each R 110 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl;

[0415] Each R 120 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; and

[0416] wherein "*" and the wavy line represent covalent bonds.

[0417] In certain embodiments, the linking moiety has the formula:

[0418]

[0419]

[0420]

[0421]

[0422]

[0423]

[0424]

[0425]

[0426] Wherein, "*" and the wavy line or the dotted line represent covalent bonds. It should be understood that either end can be connected to A 1 . In certain embodiments, the linking moiety has the following formula:

[0427]

[0428]

[0429]

[0430]

[0431]

[0432]

[0433] Wherein, "*" and the wavy line or the dotted line represent covalent bonds. It should be understood that "*", the wavy line or the dotted line can be connected to A 1 .

[0434] In certain embodiments, there is provided a compound as in Table 1, or a stereoisomer, a mixture of stereoisomers, a hydrate, a solvate, an isotope-enriched analogue or a pharmaceutically acceptable salt thereof.

[0435] Table 1

[0436]

[0437]

[0438]

[0439]

[0440]

[0441]

[0442]

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449] Treatment method

[0450] The present invention provides compounds that can be used to treat, prevent and / or delay the onset and / or development of cancer. Thus, in certain embodiments, a method for treating cancer is provided, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound or composition described herein. Certain embodiments provide a method for enhancing a cytotoxic cancer therapy in a subject recognized to be in need of such treatment, the method comprising administering to the subject a therapeutically acceptable amount of a compound or composition described herein.

[0451] It is contemplated that patients suffering from any cancer may benefit from treatment with the compounds and compositions described herein. Thus, in certain embodiments, the cancer is liver cancer, melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphoblastic leukemia, chronic lymphocytic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, nephroblastoma, cervical cancer, testicular cancer, soft tissue sarcoma, chronic lymphocytic leukemia, Waldenström macroglobulinemia, primary macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, malignant melanoma, small cell lung cancer, gastric cancer, colon cancer, malignant pancreatic insulinoma, malignant carcinoid, malignant melanoma, choriocarcinoma, mycosis fungoides, head and neck cancer, osteogenic sarcoma, pancreatic cancer, acute myeloid leukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma, urogenital cancer, thyroid cancer, esophageal cancer, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, essential thrombocythemia, adrenocortical carcinoma, skin cancer, trophoblastic tumor or prostate cancer. In certain embodiments, the cancer is bladder cancer, blood cancer (such as leukemia (e.g., chronic leukemia, chronic lymphocytic leukemia (CLL, etc.)) or lymphoma (e.g., Hodgkin lymphoma, non-Hodgkin lymphoma, low-grade lymphoma, high-grade lymphoma), lung cancer (e.g., small cell lung cancer), breast cancer, fallopian tube cancer, glioblastoma multiforme, head and neck cancer, esophageal cancer, ovarian cancer, pancreatic cancer, peritoneal cancer, prostate cancer, testicular cancer, skin cancer (e.g., melanoma) or uterine cancer. In certain embodiments, the cancer is bladder cancer, breast cancer, fallopian tube cancer, ovarian cancer, prostate cancer, peritoneal cancer, testicular cancer, endometrial cancer or uterine cancer.

[0452] In certain embodiments, the cancer is chronic lymphocytic leukemia (CLL), Hodgkin lymphoma, non-Hodgkin lymphoma, Waldenström macroglobulinemia, polycythemia vera, trophoblastic tumor, and ovarian cancer.

[0453] In certain embodiments, the compounds and compositions described herein are customized to target cancers that overexpress a specific receptor (such as, but not limited to, androgen receptor, estrogen receptor, progesterone receptor, and / or glucocorticoid receptor) by including an epitope that targets the specific nuclear receptor. The epitope can be derived from a steroid hormone or any non-steroid drug that targets the specific receptor.

[0454] Composition

[0455] The present disclosure encompasses compositions of any of the compounds detailed herein, including pharmaceutical compositions. Accordingly, there are provided pharmaceutical compositions comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient. The pharmaceutical compositions provided herein can take the form suitable for oral, buccal, parenteral (e.g., intravenous, intramuscular, infusion, or subcutaneous injection), nasal, topical, or rectal administration, or in a form suitable for administration by inhalation.

[0456] Kit

[0457] There are provided kits for effecting an anti-cancer effect, the kits comprising a compound or composition described herein. In certain embodiments, the kit comprises a unit dose of a compound or composition described herein and instructions for its administration. In certain aspects, the kit further comprises a second drug suitable for anti-cancer therapy or instructions for co-administering additional anti-cancer therapies (such as radiation or gene therapy). In another aspect, a kit for effecting an anti-cancer effect comprises a low dose (e.g., less than about 500 mg / day, or less than about 400 mg / day, or less than about 300 mg / day, or less than about 200 mg / day) of a compound or composition described herein and a second drug suitable for anti-cancer therapy. In yet another variant, a kit for effecting an anti-cancer effect comprises a high dose (e.g., greater than about 500 mg / day) of a compound or composition as described herein and a second drug suitable for anti-cancer therapy.

[0458] Method for preparing a drug

[0459] In another aspect of the present disclosure, there is provided the use of the compounds and compositions described herein in the preparation of medicaments. In particular, there is provided the preparation of a medicament for the treatment of cancer or a disease or disorder that can be mediated, at least in part, by blocking DNA repair and / or transcriptional activation (such as by inhibiting one or more topoisomerases). In addition, the pharmaceutical compositions of the compounds described herein are also intended for the preparation of a medicament for the treatment of a disease or disorder that can be mediated, at least in part, by inhibiting one or more topoisomerases.

[0460] Examples

[0461] The present disclosure is further illustrated by the following examples. The following examples represent only various aspects of the present disclosure non - restrictively. The solid and dashed wedge lines within the structures disclosed herein illustrate relative stereochemistry, where absolute stereochemistry is depicted only when explicitly stated or labeled.

[0462] Compounds having the structure of any of the compounds, formulas or any sub - formulas described herein can be synthesized using standard synthetic techniques known to those skilled in the art. The compounds of the present disclosure can be synthesized using the general synthetic methods set forth in the general methods or synthetic examples.

[0463] In cases where a particular enantiomer of a compound is desired, this can be achieved from a mixture of the corresponding enantiomers using any suitable conventional operation for the separation or resolution of enantiomers. Thus, for example, diastereomeric derivatives can be produced by the reaction of a mixture of enantiomers (e.g., a racemate) with a suitable chiral compound. The diastereomers can then be separated by any convenient means (e.g., by crystallization and recovery of the desired enantiomer). In another resolution method, racemates can be separated using chiral high - performance liquid chromatography. Alternatively, if desired, a specific enantiomer can be obtained by using a suitable chiral intermediate in one of the methods.

[0464] Chromatography, recrystallization and other conventional separation operations can also be used with intermediates or final products where a particular isomer of a compound is desired or the reaction product is otherwise purified.

[0465] Example S1: Preparation of (S) - 10 - ((4 - (1 - (6 - (((1r,4r) - 4 - (3 - chloro - 4 - cyanophenoxy) cyclohexyl) carbamoyl) pyridazin - 3 - yl) piperidin - 4 - carbonyl) piperazin - 1 - yl) methyl) - 4 - ethyl - 4 - hydroxy - 3,14 - dioxo - 3,4,12,14 - tetrahydro - 1H - pyrano[3',4':6,7] indazino[1,2 - b] quinoline - 9 - yl diisopropylcarbamate (Compound No. 1)

[0466]

[0467] Step 1: Preparation of tert-butyl (S)-4-((9-((diisopropylcarbamoyl)oxy)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazine-1-carboxylate (Intermediate A1)

[0468] To a stirred solution of tert-butyl (S)-4-((4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazine-1-carboxylate (Intermediate 3, 300 mg, 0.53 mmol, 1.0 equiv) in DCM (5 mL) was added DIPEA (0.5 mL, 3.8 mmol, 5 equiv) and DMAP (20 mg, 0.16 mmol, 0.25 equiv). Subsequently, diisopropylcarbamoyl chloride (SM-1, 132 mg, 0.80 mmol, 1.5 equiv) in DCM (2 mL) was added at room temperature and the mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (100 mL) was added and the aqueous reaction mixture was extracted with DCM (2 × 200 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by a combiflash column eluting with 4% methanol / DCM to give the product as a pale yellow solid (Intermediate A1, 240 mg, 65%).

[0469] 1 1H NMR (400 MHz, DMSO-d6) δ 9.02 (s, 1H), 8.14 - 8.16 (m, 1H), 7.60 - 7.63 (m, 1H), 7.38 (s, 1H), 6.54 (s, 1H), 5.46 (s, 2H), 5.32 (s, 2H), 3.88 (s, 2H), 3.26 - 3.42 (m, 7H), 2.40 - 2.45 (m, 3H), 1.84 - 1.92 (m, 2H), 1.39 (s, 9H), 1.33 (d, J = 7.5 Hz, 12H), 1.09 (t, J = 7.09 Hz, 3H). LCMS: 690.7 [M + H] + 。

[0470] Step 2: Preparation of (S)-4-Ethyl-4-hydroxy-3,14-dioxo-10-(piperazin-1-ylmethyl)-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl Diisopropylcarbamate Trifluoroacetate (Intermediate A2)

[0471] Under a nitrogen atmosphere at 0 °C, trifluoroacetic acid (TFA, 0.2 mL, 1.05 mmol, 3 eq) was added to a stirred solution of tert-butyl (S)-4-((9-((diisopropylcarbamoyl)oxy)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazine-1-carboxylate (Intermediate A1, 240 mg, 0.35 mmol, 1.0 eq) in DCM (5 mL). The reaction mixture was stirred and warmed to room temperature over 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, washed with diethyl ether (20 mL) and dried in vacuo to afford Intermediate A2 as an off-white solid (220 mg, 94%).

[0472] 1 H NMR (400 MHz, DMSO-d6) δ 9.02 (s, 1H), 8.52 (br s, 2H), 8.13 (d, J = 9.29 Hz, 1H), 7.58 (d, J = 9.29 Hz, 1H), 7.34 (s, 1H), 6.50 (br s, 1H), 5.44 (s, 2H), 5.31 (s, 2H), 4.23 (brs, 2H), 3.90 - 3.96 (m, 4H), 3.05 - 3.15 (m, 4H), 2.62 - 2.67 (m, 2H), 1.81 - 1.95 (m, 2H), 1.37 (d, J = 6.85 Hz, 6H), 1.29 (d, J = 6.36 Hz, 6H), 0.84 - 0.97 (d, J = 8.5 Hz, 3H). LCMS: 590.2 [M+H] + 。

[0473] Step 3: Preparation of (S)-10-((4-(1-(6-(((1r,4r)-4-(3-Chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl Diisopropylcarbamate

[0474] At room temperature, HATU (194 mg, 0.51 mmol, 1.5 equiv) and DIPEA (0.09 mL, 0.66 mmol, 2 equiv) were added to a stirred solution of intermediate A2 (200 mg, 0.33 mmol, 1.0 equiv) and intermediate 13 (CAS Registry No. 2740523-67-3; 164 mg, 0.33 mmol, 1.0 equiv) in DMF (5 mL). The resulting reaction mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (100 mL) was added and the aqueous mixture was extracted with ethyl acetate (2 × 50 mL). The combined organic extracts were washed with water (100 mL), brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The crude material was purified by combiflash column chromatography eluting with 3% methanol / DCM to give the title compound as a pale yellow solid (125 mg, 48%).

[0475] 1 1H NMR (400 MHz, DMSO-d6) δ 9.10 (br s, 1H), 8.57 (d, J = 8.25 Hz, 1H), 8.16 - 8.36 (m, 1H), 7.79 - 7.88 (m, 2H), 7.68 (br dd, J = 6.63, 4.50 Hz, 1H), 7.35 - 7.40 (m, 3H), 7.13 (dd, J = 8.82, 2.44 Hz, 1H), 6.39 - 6.68 (m, 1H), 5.44 (s, 2H), 5.35 (s, 2H), 4.46 - 4.56 (m, 3H), 4.26 - 4.35 (m, 4H), 3.78 - 3.96 (m, 4H), 3.33 - 3.63 (m, 3H), 2.92 - 3.20 (m, 4H), 2.04 - 2.16 (m, 2H), 1.84 - 1.97 (m, 4H), 1.46 - 1.77 (m, 9H), 1.33 (br dd, J = 12.57, 6.32 Hz, 12H), 0.90 (t, J = 7.32 Hz, 3H). LCMS: 1055.4 [M+H] + . HPLC purity 98.9%.

[0476] Example S2: Preparation of methyl (phenyl)carbamate (S)-10-((4-(1-(6-(((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-carbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (Compound No. 2)

[0477]

[0478] Step 1: Preparation of tert-butyl (S)-4-((4-ethyl-4-hydroxy-9-((methyl(phenyl)carbamoyl)oxy)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazine-1-carboxylate (Intermediate A3)

[0479] At room temperature, to a stirred solution of tert-butyl (S)-4-((4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazine-1-carboxylate (Intermediate 3, 300 mg, 0.53 mmol, 1.0 equiv) in DCM (15 mL) was added DIPEA (0.3 mL, 1.60 mmol, 3 equiv) and DMAP (20 mg, 0.16 mmol, 0.25 equiv), and then methyl(phenyl)carbamoyl chloride (SM-1, 135 mg, 0.80 mmol, 1.5 equiv) in DCM (5 mL) was added. Subsequently, the resulting reaction mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted in water (100 mL) and extracted with DCM (2 × 200 mL). The combined organic extracts were washed with brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash column using 6% methanol / DCM to give Intermediate A3 as an off-white solid (260 mg, 70%).

[0480] 1 H NMR (400 MHz, DMSO-d6) δ 8.96 (br s, 1H), 8.06 - 8.19 (m, 2H), 7.74 (d, J = 9.13 Hz, 1H), 7.55 (br d, J = 7.63 Hz, 1H), 7.44 - 7.46 (m, 1H), 7.28 - 7.35 (m, 2H), 6.57 - 6.62 (m, 1H), 6.48 - 6.54 (m, 1H), 5.42 (s, 2H), 5.29 (s, 2H), 3.69 - 3.82 (m, 1H), 3.38 - 3.40 (m, 3H), 3.16 - 3.19 (m, 3H), 2.95 (s, 3H), 2.65 (d, J = 5.13 Hz, 1H), 2.12 - 2.27 (m, 3H), 1.85 - 1.90 (m, 1H), 1.41 (s, 9H), 0.88 (t, J = 7.32 Hz, 3H). LCMS: 696.2 [M + H]+ 。

[0481] Step 2: Preparation of (S)-4-Ethyl-4-hydroxy-3,14-dioxo-10-((piperazin-1-ylmethyl)-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) methyl(phenyl)carbamate trifluoroacetate (Intermediate A4)

[0482] Under a nitrogen atmosphere at 0 °C, trifluoroacetic acid (TFA, 0.27 mL, 3.5 mmol, 10 eq) was added to a stirred solution of tert-butyl (S)-4-((4-ethyl-4-hydroxy-9-((methyl(phenyl)carbamoyl)oxy)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazine-1-carboxylate (Intermediate A3, 250 mg, 0.35 mmol, 1.0 eq) in DCM (10 mL). The reaction mixture was allowed to warm to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, washed with diethyl ether (20 mL), and dried in vacuo to give Intermediate A4 (210 mg, 98%) as an off-white solid.

[0483] 1 H NMR (400 MHz, DMSO-d6) δ 8.99 (br s, 1H), 8.56 (br s, 2H), 8.15 (br d, J = 9.29 Hz, 1H), 7.75 (br d, J = 8.80 Hz, 1H), 7.53 - 7.58 (m, 3H), 7.49 (br t, J = 7.58 Hz, 1H), 7.34 (s, 1H), 6.50 (br s, 1H), 5.43 (s, 2H), 5.29 (br s, 2H), 3.81 - 3.96 (m, 3H), 3.42 (br s, 3H), 2.98 - 3.01 (m, 5H), 2.52 - 2.56 (m, 3H), 1.80 - 1.96 (m, 2H), 0.89 (br t, J = 7.09 Hz, 3H).

[0484] Step 3: Preparation of (S)-10-((4-(1-(6-(((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl methyl(phenyl)carbamate

[0485] At room temperature, HATU (240 mg, 0.67 mmol, 1.5 equiv) and DIPEA (0.17 mL, 1 mmol, 3 equiv) were added to a stirred solution of methyl (phenyl)carbamate (S)-4-ethyl-4-hydroxy-3,14-dioxo-10-(piperazin-1-ylmethyl)-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl ester trifluoroacetate (Intermediate A4, 200 mg, 0.33 mmol, 1.0 equiv) and 1-(6-(((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carboxylic acid (Intermediate 13, 200 mg, 0.40 mmol, 1.2 equiv) in DMF (5 mL). The resulting reaction mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted in water (100 mL) and extracted with 10% methanol / DCM (2 x 50 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by preparative HPLC (in ammonium bicarbonate / water / acetonitrile mobile phase) to give the title compound as an off-white solid (75 mg, 21%).

[0486] 1 H NMR (400 MHz, DMSO-d6) δ 8.99 (br s, 1H), 8.59 (d, J = 8.25 Hz, 1H), 8.14 (d, J = 9.13 Hz, 1H), 7.84 (dd, J = 10.94, 9.19 Hz, 2H), 7.76 (d, J = 9.26 Hz, 1H), 7.55 (br d, J = 7.75 Hz, 2H), 7.47 (t, J = 7.75 Hz, 2H), 7.31 - 7.41 (m, 4H), 7.13 (dd, J = 8.76, 2.38 Hz, 1H), 6.51 (s, 1H), 5.43 (s, 2H), 5.30 (s, 2H), 4.44 - 4.57 (m, 3H), 3.72 - 3.94 (m, 3H), 3.42 (s, 3H), 3.38 - 3.41 (m, 4H), 3.07 - 3.18 (m, 2H), 2.91 - 3.01 (m, 1H), 2.07 - 2.29 (m, 5H), 1.80 - 1.97 (m, 4H), 1.43 - 1.76 (m, 9H), 0.89 (t, J = 7.32 Hz, 3H). LCMS: 1061.4 [M+H] + . HPLC purity 98.2%.

[0487] Example S3: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl 3-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (Compound No. 3)

[0488]

[0489] Step 1: Preparation of tert-butyl 6-(chlorocarbonyl)-3,6-diazabicyclo[3.1.1]heptane-3-carboxylate (Intermediate 14)

[0490] To a stirred solution of tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate (SM-1, 1.0 g, 5.05 mmol, 1.0 equiv) in DCM (10 mL) at 0 °C was added dropwise a solution of pyridine (0.81 mL, 10.10 mmol, 2 equiv) and triphosgene (0.45 g, 1.51 mmol, 0.3 equiv) in DCM (5 mL) over 10 min. The reaction mixture was warmed to room temperature and stirred for 2 h. The progress of the reaction was monitored by TLC (non-polar spots were observed). After completion of the reaction, the reaction mixture was poured into ice water (20 mL) and extracted with DCM (2 x 15 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give Intermediate 14 (1.0 g, crude) as a pale yellow solid, which was used in the next step without further purification.

[0491] 1 H NMR (400 MHz, DMSO-d6) δ 4.63 - 4.76 (m, 2H), 3.57 - 3.84 (m, 4H), 2.91 - 2.99 (m, 1H), 1.70 - 1.78 (m, 1H), 1.44 (s, 9H).

[0492] Step 2: Preparation of 3-(tert-butyl) 3,6-diazabicyclo[3.1.1]heptane-3,6-dicarboxylate · 6-((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 15)

[0493] At 0 °C, DIPEA (2.12 mL, 11.87 mmol, 5 equiv) was added to a stirred solution of tert-butyl 6-(chlorocarbonyl)-3,6-diazabicyclo[3.1.1]heptane-3-carboxylate (Intermediate 16, 0.92 g, 3.56 mmol, 1.5 equiv) and (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-2, 1 g, 2.37 mmol, 1 equiv) in THF (10 mL) and DMF (10 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with water (20 mL) and extracted with ethyl acetate (2 × 20 mL). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product. The obtained crude product was purified by combiflash column chromatography eluting with 6% methanol / DCM to give Intermediate 15 (650 mg, 42%) as a pale yellow solid.

[0494] 1 H NMR (400 MHz, DMSO-d6) δ 8.96 (s, 1H), 8.10 (d, J = 8.63 Hz, 1H), 7.51 (d, J = 9.01 Hz, 1H), 7.34 (s, 1H), 6.51 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 3.70 - 3.87 (m, 2H), 3.55 - 3.66 (m, 6H), 2.69 - 2.77 (m, 1H), 2.12 - 2.24 (m, 6H), 1.81 - 1.93 (m, 2H), 1.54 - 1.59 (m, 1H), 1.47 (s, 9H), 0.89 (t, J = 7.38 Hz, 3H). LCMS: 646.50 [M+H] + 。

[0495] Step 3: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate trifluoroacetate (Intermediate 16)

[0496] At 0 °C, trifluoroacetic acid (TFA, 0.57 mL, 7.55 mmol, 7.5 eq) was added to a stirred solution of 3-(tert-butyl) 3,6-diazabicyclo[3.1.1]heptane-3,6-dicarboxylate · 6-((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 17, 650 mg, 1.0 mmol, 1.0 eq) in DCM (10 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure to give the crude product, which was triturated with ethyl acetate (30 mL), filtered, and the collected solid was dried in vacuo to give Intermediate 16 as a yellow solid (500 mg, 75%).

[0497] 1 H NMR (400 MHz, D2O) δ 8.99 (s, 1H), 8.33 (d, J = 9.38 Hz, 1H), 7.95 (d, J = 9.26 Hz, 1H), 7.63 (s, 1H), 5.29 - 5.60 (m, 4H), 4.92 - 5.00 (m, 2H), 3.97 - 4.24 (m, 3H), 3.72 - 3.88 (m, 3H), 3.19 - 3.31 (m, 2H), 3.05 (s, 3H), 3.02 (s, 4H), 1.93 - 2.09 (m, 3H), 0.98 (t, J = 7.19 Hz, 3H). LCMS: 546.2 & 547.2 [M+H] + 。

[0498] Step C1: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methylamino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 17)

[0499] At 0 °C, KOAc (20.6 g, 210 mmol, 10 eq) and iodine (13.1 g, 105 mmol, 5 eq) were added to a stirred solution of SM-1 (10 g, 21 mmol, 1.0 eq) in methanol (150 mL) and THF (150 mL). The reaction mixture was warmed to room temperature and stirred for 3 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with sodium thiosulfate (Na2S2O3) (50 g in 30 mL of water) and extracted with ethyl acetate (2 × 200 mL). The combined organic extracts were washed with brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford Intermediate 17 as an off-white solid (8.0 g, 82%), which was used in the next step without further purification.

[0500] 1 H NMR (400 MHz, DMSO-d6) δ 11.91 (br s, 1H), 6.91 (d, J = 8.31 Hz, 2H), 6.44 (d, J = 8.31 Hz, 2H), 5.67 (s, 1H), 4.37 (m, 1H), 2.75 (s, 2H), 2.61 (d, J = 4.40 Hz, 3H), 2.30 - 2.40 (m, 1H), 2.07 - 2.16 (s, 5H), 1.99 (s, 6H), 1.63 - 1.77 (m, 2H), 1.21 - 1.45 (m, 5H), 0.86 (t, J = 6.60 Hz, 1H), 0.16 - 0.28 (m, 3H). LCMS: 462.28 [M+H] + 。

[0501] Step C2: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((6-hydroxyhexyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 18)

[0502] At room temperature, to a solution of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methylamino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 17, 4 g, 8.67 mmol, 1.0 equiv) and 6-bromohexan-1-ol (SM-2, 7.81 g, 43.38 mmol, 5 equiv) in ethanol (40 mL) and water (40 mL) was added NaHCO3 (7.37 g, 86.76 mmol, 10 equiv). The reaction mixture was heated to 80 °C and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through a pad of diatomaceous earth and washed with ethyl acetate (40 mL). The filtrate was concentrated under reduced pressure, diluted with water (120 mL) and extracted with ethyl acetate (2 × 200 mL). The combined organic extracts were washed with brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash chromatography eluting with 70% ethyl acetate / heptane to give Intermediate 18 as an off-white solid (2.6 g, 53%).

[0503] 1 H NMR (400 MHz, DMSO-d6) δ 6.98 (d, J = 7.89 Hz, 2H), 6.58 (d, J = 7.89 Hz, 2H), 5.67 (br s, 1H), 4.24 - 4.51 (m, 2H), 3.36 (d, J = 5.70 Hz, 2H), 3.23 (d, J = 6.58 Hz, 2H), 2.69 - 2.86 (m, 4H), 2.55 (s, 3H), 2.29 - 2.44 (m, 1H), 2.05 - 2.26 (m, 5H), 1.87 - 2.04 (m, 6H), 1.63 - 1.77 (m, 2H), 1.34 - 1.49 (m, 6H), 1.27 (br s, 6H), 0.23 (br s, 3H). LCMS: 562.40 [M+H] + 。

[0504] Step C3: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(6-oxohexyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate C)

[0505] At 0 °C, Dess-Martin periodinane (DMP) (1.1 g, 2.67 mmol, 3 eq) was added portionwise to a stirred solution of Intermediate 18 (500 mg, 0.891 mmol, 1 eq) in ethyl acetate (40 mL). The reaction mixture was heated to 80 °C and maintained for 2 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with 50% aqueous Na2S2O3 solution (10 mL), saturated NaHCO3 solution (15 mL) and extracted with ethyl acetate (2 X 25 mL). The combined organic extracts were washed with brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give Intermediate C (450 mg, 92%) as a brown solid.

[0506] 1 H NMR (400 MHz, DMSO-d6) δ 9.64 (s, 1H), 6.98 (d, J = 8.31 Hz, 2H), 6.58 (d, J = 8.80 Hz, 2H), 5.67 (s, 1H), 4.39 (d, J = 5.87 Hz, 1H), 3.22 (t, J = 6.60 Hz, 2H), 2.55 - 2.80 (m, 5H), 2.51–2.54 (m, 2H), 2.40 (t, J = 7.09 Hz, 2H), 1.96 - 2.15 (m, 12H), 1.56 - 1.69 (m, 2H), 1.11 - 1.59 (m, 10H), 0.23 (s, 3H). LCMS: 560.4 [M+H] + 。

[0507] Step 4: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 3-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate

[0508] At room temperature, glacial acetic acid (0.1 mL) was added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(6-oxohexyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate C, 350 mg, 0.531 mmol, 1 equiv) and (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate trifluoroacetate (Intermediate 16, 296 mg, 0.531 mmol, 1 equiv) in methanol (7 mL), and the mixture was stirred for an additional 2 h. Then, sodium cyanoborohydride (65.8 mg, 1.06 mmol, 2 equiv) was added at 0 °C, and the reaction mixture was warmed to room temperature and stirred for 16 h. The reaction progress was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, quenched with ice water (20 mL), and extracted with 10% methanol / DCM (2 x 20 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product. The obtained crude product was purified by preparative HPLC eluting with mobile phase A: 0.1% FA / water and mobile phase B: acetonitrile to afford the title compound as an off-white solid (20 mg, 3%).

[0509] 11H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.13 (d, J = 9.26 Hz, 1H), 7.70 (d, J = 9.13 Hz, 1H), 7.35 (s, 1H), 6.97 (d, J = 8.63 Hz, 2H), 6.58 (d, J = 8.63 Hz, 2H), 6.52 (s, 1H), 5.62 - 5.70 (m, 1H), 5.43 (s, 2H), 5.32 (s, 2H), 4.35 - 4.43 (m, 1H), 3.88 - 3.97 (m, 1H), 3.72 - 3.85 (m, 2H), 3.52 - 3.71 (m, 4H), 3.34 - 3.42 (m, 1H), 3.21 - 3.28 (m, 2H), 2.82 (s, 3H), 2.65 - 2.79 (m, 2H), 2.54 - 2.64 (m, 2H), 2.28 - 2.39 (m, 1H), 2.19 (s, 6H), 2.11 - 2.17 (m, 3H), 2.09 (s, 3H), 1.96 - 2.01 (m, 4H), 1.83 - 1.93 (m, 3H), 1.63 - 1.77 (m, 2H), 1.54 - 1.61 (m, 1H), 1.42 - 1.53 (m, 2H), 1.21 - 1.41 (m, 11H), 0.89 (t, J = 7.32 Hz, 3H), 0.22 (s, 3H), -0.06 (s, 2H). LCMS: 1089.57 [M + H] + . HPLC purity 86.1%

[0510] Example S4: Preparation of 2-(2-(((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)-N-methylacetamido)-N-(((S)-4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)-N,2-dimethylpropanamide (Compound No. 4)

[0511]

[0512] Step 1: Preparation of 2-methyl-2-(methylamino)propanoic acid trifluoroacetate (Intermediate 19)

[0513] Under a nitrogen atmosphere, at 0 °C, trifluoroacetic acid (TFA, 1.7 mL) was added to a stirred solution of 2-((tert-butoxycarbonyl)(methyl)amino)-2-methylpropanoic acid (SM-1, 500 mg, 2.30 mmol, 1.0 equiv) in DCM (10 mL). The reaction mixture was warmed to room temperature and stirred for 3 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, the reaction mixture was washed with diethyl ether (2 × 20 mL) and dried under vacuum to give Intermediate 19 as an off-white solid (450 mg, 93%).

[0514] 1 1H NMR (400 MHz, DMSO-d6) δ 14.125 (br s, 1H), 9.09 (br s, 2H), 2.51 (d, J = 11.25 Hz, 3H), 1.40 (s, 6H). LCMS: 118.08 [M+H] + 。

[0515] Step 2: Preparation of (S)-N-((4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]-indazino[1,2-b]quinolin-10-yl)methyl)-N,2-dimethyl-2-(methylamino)propanamide (Intermediate 20)

[0516] At room temperature, HATU (703 mg, 1.96 mmol, 2.0 equiv) and DIPEA (0.5 mL, 2.94 mmol, 3 equiv) were added to a stirred solution of (S)-4-ethyl-4,9-dihydroxy-10-((methylamino)methyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione (Intermediate 2, 400 mg, 0.98 mmol, 1.0 equiv) and 2-methyl-2-(methylamino)propanoic acid trifluoroacetate (Intermediate 19, 137 mg, 1.17 mmol, 1.2 equiv) in DMF (2 mL), and the resulting reaction mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was washed with water (60 mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give a crude product. The obtained crude product was purified by combiflash column eluting with 6% methanol / DCM to give Intermediate 20 as an off-white solid (210 mg, 42%).

[0517] 11H NMR (400 MHz, DMSO-d6) δ 8.43 (br s, 1H), 8.27 (br d, J = 7.83 Hz, 2H), 8.04 (br d, J = 8.80 Hz, 1H), 7.60 (br d, J = 9.29 Hz, 1H), 7.18 - 7.36 (m, 1H), 6.50 (br s, 1H), 5.41 (br s, 2H), 5.23 (br s, 2H), 5.09 (br s, 2H), 3.33 - 3.40 (m, 2H), 3.00 (br s, 3H), 2.33 (br s, 3H), 1.40 (s, 6H), 0.87 (t, J = 7.32 Hz, 3H). LCMS: 507.45 [M+H] + 。

[0518] Step 3: Preparation of 2-(2-(((5S,8R,9S,10S,13S,14S,17S)-10,13-Dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)-N-methylacetamido)-N-(((S)-4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)-N,2-dimethylpropanamide

[0519] At room temperature, HATU (212 mg, 0.59 mmol, 2.0 equiv) and DIPEA (0.15 mL, 0.88 mmol, 3 equiv) were added to a stirred solution of (S)-N-((4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)-N,2-dimethyl-2-(methylamino)acrylamide (Intermediate 20, 150 mg, 0.29 mmol, 1.0 equiv) and 2-(((5S,8R,9S,10S,13S,14S,17S)-10,13-dimethyl-3-oxohexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)acetic acid Intermediate 4 (113 mg, 0.32 mmol, 1.1 equiv) in DMF (2 mL). The reaction mixture was heated to 50 °C and stirred for 4 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (10 mL) was added and the aqueous reaction mixture was extracted with ethyl acetate (2 × 100 mL). The combined organic extracts were washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by preparative HPLC (in ammonium bicarbonate / water / acetonitrile mobile phase) to give the title compound as an off-white solid (7 mg, 3%).

[0520] 1 1H NMR (400 MHz, DMSO-d6) δ 10.50 (d, J = 4.38 Hz, 1H), 8.57 (s, 1H), 7.99 (d, J = 9.13 Hz, 1H), 7.52 (d, J = 9.13 Hz, 1H), 7.27 (s, 1H), 6.46 (s, 1H), 5.41 (br s, 2H), 5.30 (br s, 2H), 5.09 (br d, J = 12.38 Hz, 1H), 4.90 (br d, J = 12.38 Hz, 1H), 3.79 - 3.93 (m, 2H), 2.81 (s, 3H), 2.59 (s, 3H), 2.25 (br d, J = 14.13 Hz, 1H), 2.12 (br d, J = 15.26 Hz, 1H), 1.79 - 1.93 (m, 4H), 1.48 - 1.50 (m, 3H), 1.45 (s, 6H), 1.21 - 1.33 (m, 4H), 1.06 - 1.20 (m, 4H), 0.92 - 1.03 (m, 2H), 0.91 (s, 3H), 0.75 - 0.90 (m, 6H), 0.40 - 0.52 (m, 1H), 0.22 - 0.39 (m, 2H), 0.17 (s, 3H). LCMS: 837.35 [M + H] + 。The HPLC purity is 98.8%.

[0521] Example S5: Preparation of (S)-4-ethyl-4-hydroxy-10-((4-methylpiperazin-1-yl)methyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylate (Compound No. 5)

[0522]

[0523] Step 1: Preparation of (S)-4-ethyl-4,9-dihydroxy-10-((4-methylpiperazin-1-yl)methyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione (Intermediate 1)

[0524] Under ambient temperature and argon atmosphere, formaldehyde (3.95 mL, 41.20 mmol, 1.5 equiv, 37%, in water) and 1-methylpiperazine (SM-2, 4.12 g, 41.20 mmol, 1.5 equiv) were added to a stirred solution of (S)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indolo[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-1, 10 g, 27.47 mmol, 1.0 equiv) in AcOH (100 mL, 10 vol). The resulting reaction mixture was stirred at 80 °C until TLC indicated complete consumption of the starting material. Then the reaction mixture was diluted with ice water (250 mL), extracted with 10% MeOH / DCM (2 x 250 mL), the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The product was washed with ether (100 mL) and dried to give (S)-4-ethyl-4,9-dihydroxy-10-((4-methylpiperazin-1-yl)methyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione (Intermediate 1, 6 g, 46%) as an off-white solid.

[0525] 1 H NMR (400 MHz, DMSO-d6) δ 8.73 (s, 1H), 7.95 - 8.00 (m, 1H), 7.39 - 7.44 (m, 1H), 7.24 - 7.27 (m, 1H), 6.43 - 6.52 (m, 1H), 5.39 - 5.44 (m, 2H), 5.22 - 5.26 (m, 2H), 4.07 - 4.12 (m, 2H), 2.72 - 2.78 (m, 1H), 2.57 - 2.64 (m, 2H), 2.25 - 2.38 (m, 4H), 2.12 - 2.19 (m, 4H), 1.82 - 1.91 (m, 3H), 0.85 - 0.92 (m, 3H). LCMS: 477.3 [M+H] + 。

[0526] Step 2: Preparation of (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate · 4-(4-ethyl-4-hydroxy-10-((4-methylpiperazin-1-yl)methyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2)

[0527] At 0 °C under an argon atmosphere, pyridine (5.47 mL, 67.2 mmol, 2.5 equiv) and triphosgene (3.99 g, 13.44 mmol, 0.5 equiv) were added to a stirred solution of tert-butyl piperazine-1-carboxylate (SM-3, 5 g, 26.88 mmol, 1.0 equiv) in DCM (50 mL, 10 vol). The reaction mixture was warmed to room temperature and stirred for 2 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with 1N HCl (~200 mL) and extracted with DCM (2 × 200 mL). The combined organic layers were washed with an aqueous sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give crude 4-(chlorocarbonyl)piperazine-1-carboxylic acid tert-butyl ester as a semi-solid. At 0 °C under an argon atmosphere, DIPEA (6.19 mL, 33.6 mmol, 2.5 equiv) and DMAP (327 mg, 2.68 mmol, 0.1 equiv) were added to a second flask containing (S)-4-ethyl-4,9-dihydroxy-10-((4-methylpiperazin-1-yl)methyl)-1,12-dihydro-14H-pyrano[3',4':6,7]-indazino[1,2-b]quinoline-3,14(4H)-dione (Intermediate 1, 6.3 g, 13.44 mmol, 0.5 equiv) in DCM (25 mL, 10 vol). The above carbonyl chloride was dissolved in DCM (25 mL) and added dropwise to the reaction mixture at 0 °C under an argon atmosphere. Subsequently, the resulting reaction mixture was stirred at ambient temperature until TLC indicated complete consumption of the starting material. Then the reaction mixture was concentrated under reduced pressure, diluted with ice water (100 mL), the resulting precipitate was filtered and the crude product was dried and purified by flash column (silica gel, 100 - 200 mesh) eluting with 2 - 5% MeOH / DCM. The pure fractions were combined and concentrated under reduced pressure to give (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate · 4-(4-ethyl-4-hydroxy-10-((4-methylpiperazin-1-yl)methyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]-indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2, 1.56 g, 26%) as an off-white solid.

[0528] 1 H NMR (400 MHz, DMSO-d6) δ 8.98 (s, 1H), 8.12 (d, J = 9.29 Hz, 1H), 7.65 (d, J = 9.17 Hz, 1H), 7.34 (s, 1H), 5.75 (s, 1H), 5.42 (s, 2H), 5.30 (s, 2H), 3.66

[0529] -3.88 (m, 4H), 3.40 - 3.56 (m, 6H), 2.51 - 2.57 (m, 2H), 2.46 - 2.49 (m, 1H), 2.14

[0530] -2.29 (m, 4H), 1.85 - 1.91 (m, 1H), 1.82 - 1.93 (m, 1H), 1.40 - 1.46 (m, 11H), 0.89 (t, J = 7.27 Hz, 3H). LCMS: 689.4 [M+H] + 。

[0531] Step 3: Preparation of (S)-4-Ethyl-4-hydroxy-10-((4-methylpiperazin-1-yl)methyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate 3)

[0532] At ambient temperature, trifluoroacetic acid (TFA) (1.66 mL, 21.8 mmol, 10 eq) was added to a stirred solution of piperazine-1,4-dicarboxylic acid (S)-1-(tert-butyl) ester·4-(4-ethyl-4-hydroxy-10-((4-methylpiperazin-1-yl)methyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2, 1.5 g, 2.18 mmol, 1.0 eq) in DCM (15 mL, 10 vol). The reaction mixture was stirred at ambient temperature until TLC indicated complete consumption of the starting material. The reaction mixture was then concentrated under reduced pressure, saturated sodium bicarbonate solution (100 mL) was added, and the aqueous mixture was extracted with 10% MeOH / DCM (2 × 250 mL). The combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure and triturated with ether (100 mL) to afford (S)-4-ethyl-4-hydroxy-10-((4-methylpiperazin-1-yl)methyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate as an off-white solid.

[0533] 11H NMR (400 MHz, DMSO-d6) δ 8.91 - 8.99 (m, 1H), 8.04 - 8.13 (m, 1H), 7.59 - 7.64 (m, 1H), 7.30 - 7.35 (m, 1H), 6.48 - 6.53 (m, 1H), 5.38 - 5.46 (m, 2H), 5.26 - 5.33 (m, 2H), 3.81 - 3.86 (m, 2H), 3.56 - 3.68 (m, 3H), 3.36 - 3.42 (m, 2H), 3.12 - 3.18 (m, 2H), 2.74 - 2.85 (m, 5H), 2.16 - 2.34 (m, 4H), 2.07 - 2.13 (m, 4H), 1.81 - 1.92 (m, 2H), 0.83 - 0.91 (m, 3H). LCMS: 589.2 [M+H] + 。

[0534] Step 4: Preparation of (S)-4-ethyl-4-hydroxy-10-((4-methylpiperazin-1-yl)methyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylate

[0535] At 0 °C under an argon atmosphere, (S)-4-ethyl-4-hydroxy-10-((4-methylpiperazin-1-yl)methyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl piperazine-1-carboxylate (Intermediate 3, 2.0 g, 3.40 mmol, 1.0 equiv) in a stirred solution of MeOH (20 mL, 10 vol) was added (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(6-oxohexyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate C, 3.8 g, 6.80 mmol, 2.0 equiv) and acetic acid (1 mL, catalytic amount). The reaction mixture was stirred at room temperature for 1 h and NaCNBH3 (428 mg, 6.80 mmol, 2.0 equiv) was added at 0 °C under an argon atmosphere. The resulting reaction mixture was then stirred at room temperature until TLC indicated complete consumption of the starting material. The reaction mixture was then quenched with saturated sodium bicarbonate solution (250 mL), extracted with ethyl acetate (2 × 500 mL), and the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (column: spherical-C18, 40 um, 100 A; mobile phase A: 0.1% FA / water; mobile phase B: acetonitrile; flow rate: 25 mL / min, diluent: THF + DMSO). The pure fractions were lyophilized under reduced pressure to give (S)-4-ethyl-4-hydroxy-10-((4-methylpiperazin-1-yl)methyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylate as an off-white solid (486 mg, 12%).

[0536] 11H NMR (400 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.14 - 8.15 (m, 2H), 8.09 - 8.13 (m, 1H), 7.63 (d, J = 9.26 Hz, 1H), 7.34 (s, 1H), 6.96 - 7.00 (m, 2H), 6.58 - 6.62 (m, 3H), 6.48 - 6.54 (m, 2H), 5.66 - 5.69 (m, 1H), 5.42 - 5.44 (m, 2H), 5.28 - 5.33 (m, 2H), 4.38 - 4.42 (m, 1H), 4.38 - 4.42 (m, 1H), 3.82 - 3.86 (m, 2H), 3.66 - 3.72 (m, 2H), 3.44 - 3.50 (m, 4H), 3.22 - 3.27 (m, 5H), 2.29 - 2.37 (m, 7H), 2.08 - 2.18 (m, 11H), 1.42 - 1.51 (m, 5H), 1.26 - 1.35 (m, 7H), 0.86 - 0.91 (m, 3H), 0.22 - 0.25 (m, 3H). LCMS: 1133.7 [M + H] + . HPLC purity 91.5%.

[0537] Example S6: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(7-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)heptyl)piperazine-1-carboxylate (Compound No. 6)

[0538]

[0539] Step 1: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((7-hydroxyheptyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1)

[0540] At 0 °C under an argon atmosphere, 7-bromoheptanol (SM-2, 10.56 g, 54.2 mmol, 5.0 equiv) and NaHCO3 (2.73 g, 32.52 mmol, 3.0 equiv) were added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methylamino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (SM-1, 5.0 g, 10.84 mmol, 1.0 equiv) in EtOH (50 mL, 10 vol) and H2O (25 mL, 5 vol). The resulting reaction mixture was stirred at 80 °C until TLC indicated complete consumption of the starting material. The reaction mixture was then diluted with ice water (250 mL), extracted with EtOAc (2 × 500 mL), the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting crude product was purified by flash column (silica gel, 100 - 200 mesh) eluting with 20 - 50% EtOAc / hexane. The pure fractions were combined and concentrated under reduced pressure to afford (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((7-hydroxyheptyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1, 2 g, 32%) as an off-white solid.

[0541] 1H NMR (400 MHz, DMSO-d6) δ 6.97 (d, J = 8.31 Hz, 2H), 6.57 (d, J = 8.31 Hz, 2H), 5.64 - 5.70 (m, 1H), 4.36 - 4.41 (m, 1H), 4.27 - 4.32 (m, 1H), 3.33 - 3.38 (m, 3H), 3.18 - 3.26 (m, 2H), 2.78 - 2.82 (m, 3H), 2.64 - 2.77 (m, 2H), 2.54 - 2.64 (m, 2H), 2.30 - 2.41 (m, 1H), 2.19 - 2.24 (m, 1H), 2.11 - 2.15 (m, 1H), 2.06 - 2.11 (m, 3H), 1.97 - 2.03 (m, 4H), 1.85 - 1.93 (m, 1H), 1.64 - 1.75 (m, 2H), 1.35 - 1.50 (m, 6H), 1.19 - 1.32 (m, 8H), 0.17 - 0.26 (m, 3H). LCMS: 576.96 [M+H] + 。

[0542] Step 2: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(7-oxoheptyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 2)

[0543] Under an argon atmosphere at 0 °C, Dess-Martin periodinane (DMP, 4.4 g, 10.4 mmol, 2.0 equiv) was added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((7-hydroxyheptyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1, 3 g, 5.2 mmol, 1.0 equiv) in EtOAc (60 mL, 20 vol). The resulting reaction mixture was stirred at 80 °C until TLC indicated complete consumption of the starting material. Subsequently, the reaction mixture was quenched with a mixture of Na2S2O3 and saturated sodium bicarbonate solution (1:1, 250 mL), extracted with EtOAc (2 × 500 mL), and the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford crude (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(7-oxoheptyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 2, 3 g, crude) as an off-white solid, which was used in the next step without further purification.

[0544] 1 1H NMR (400 MHz, DMSO-d6) δ 9.65 (s, 1H), 6.97 (d, J = 7.82 Hz, 1H), 6.58 (d, J = 8.80 Hz, 1H), 5.67 (s, 1H), 4.39 (d, J = 6.36 Hz, 1H), 4.03 (q, J = 6.85 Hz, 1H), 3.17 - 3.26 (m, 2H), 2.65 - 2.84 (m, 4H), 2.52 - 2.65 (m, 2H), 2.28 - 2.44 (m, 3H), 2.05 - 2.25 (m, 6H), 1.84 - 2.04 (m, 7H), 1.62 - 1.76 (m, 2H), 1.34 - 1.54 (m, 5H), 1.21 - 1.31 (m, 5H), 1.15 - 1.20 (m, 2H), 0.21 - 0.26 (m, 3H). LCMS: 574.72 [M+H]+ 。

[0545] Step 3: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(7-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)heptyl)piperazine-1-carboxylate

[0546] At 0 °C under an argon atmosphere, acetic acid (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(7-oxoheptyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl ester (Intermediate 2, 3.2 g, 5.62 mmol, 1.0 equiv) and acetic acid (1 mL, catalytic amount) were added to a stirred solution of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate A, 3.0 g, 5.62 mmol, 1.0 equiv) in MeOH (30 mL, 10 vol). The reaction mixture was stirred at room temperature for 1 h, and then NaCNBH3 (705 mg, 11.24 mmol, 2.0 equiv) was added at 0 °C under an argon atmosphere. Subsequently, the resulting reaction mixture was stirred at room temperature until TLC indicated complete consumption of the starting materials. The reaction mixture was then quenched with saturated sodium bicarbonate solution (250 mL), extracted with ethyl acetate (2 × 500 mL), the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was purified by preparative HPLC (column: spherical-C18, 40 um, 100 A; mobile phase A: 0.1% FA / water; mobile phase B: acetonitrile; flow rate: 25 mL / min, diluent: THF + DMSO). The pure fractions were combined and lyophilized under reduced pressure to obtain (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl 4-(7-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)heptyl)piperazine-1-carboxylate as an off-white solid (2.35 g, 38%).

[0547] 11H NMR (400 MHz, DMSO-d6) δ 8.96 - 9.00 (m, 1H), 8.09 - 8.16 (m, 1H), 7.60 - 7.65 (m, 1H), 7.32 - 7.36 (m, 1H), 6.97 - 7.01 (m, 2H), 6.57 - 6.62 (m, 2H), 6.49 - 6.51 (m, 1H), 5.66 - 5.70 (m, 1H), 5.42 - 5.44 (m, 2H), 5.30 - 5.32 (m, 2H), 4.37 - 4.42 (m, 1H), 3.80 - 3.87 (m, 2H), 3.64 - 3.73 (m, 2H), 3.42 - 3.51 (m, 2H), 3.24 - 3.25 (m, 3H), 2.80 - 2.84 (m, 4H), 2.41 - 2.43 (m, 2H), 2.31 - 2.34 (m, 4H), 2.08 - 2.15 (m, 12H), 1.99 - 2.01 (m, 5H), 1.85 - 1.92 (m, 4H), 1.65 - 1.73 (m, 3H), 1.41 - 1.51 (m, 5H), 1.27 - 1.34 (m, 6H), 0.86 - 0.91 (m, 4H), 0.23 - 0.26 (m, 3H). LCMS: 1091.1 [M + H] + 。

[0548] Example S7: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(8-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)octyl)piperazine-1-carboxylate (Compound No. 7)

[0549]

[0550] Step 1: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((8-hydroxyoctyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1)

[0551] At 0 °C under an argon atmosphere, 8-bromooctanol (SM-2, 22.66 g, 108.45 mmol, 5.0 equiv) and NaHCO3 (5.46 g, 61.07 mmol, 3.0 equiv) were added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methylamino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (SM-1, 10.0 g, 21.69 mmol, 1.0 equiv) in EtOH (100 mL, 10 vol) and H2O (50 mL, 5 vol). The resulting reaction mixture was stirred at 80 °C until TLC indicated complete consumption of the starting material. The reaction mixture was then diluted with ice water (250 mL), extracted with EtOAc (2 × 500 mL), the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting crude product was purified by flash column (silica gel, 100 - 200 mesh) eluting with 20 - 50% EtOAc / hexanes. The pure fractions were combined and concentrated under reduced pressure to afford (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((8-hydroxyoctyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1, 5 g, 39%) as an off-white solid.

[0552] 1 H NMR (400 MHz, DMSO-d6) δ 6.97 (d, J = 7.83 Hz, 2H), 6.57 (d, J = 7.83 Hz, 2H), 5.74 - 5.77 (m, 1H), 5.65 - 5.69 (m, 1H), 4.36 - 4.42 (m, 1H), 4.26 - 4.35 (m, 2H), 3.33 - 3.40 (m, 6H), 3.18 - 3.26 (m, 1H), 2.80 (s, 3H), 2.67 - 2.77 (m, 1H), 2.54 - 2.64 (m, 2H), 2.31 - 2.41 (m, 1H), 2.12 - 2.24 (m, 2H), 2.10 (s, 3H), 1.89 - 2.04 (m, 3H), 1.61 - 1.78 (m, 1H), 1.35 - 1.45 (m, 6H), 1.21 - 1.30 (m, 10H), 0.23 (s, 3H). LCMS: 590.5 [M+H] + 。

[0553] Step 2: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(8-oxooctyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 2)

[0554] Under an argon atmosphere at 0 °C, DMP (5.75 g, 13.5 mmol, 2.0 eq) was added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((8-hydroxyoctyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1, 4 g, 6.79 mmol, 1.0 eq) in EtOAc (80 mL, 20 vol). The resulting reaction mixture was stirred at 80 °C until TLC indicated complete consumption of the starting material. Subsequently, the reaction mixture was quenched with a mixture of Na2S2O3 and saturated sodium bicarbonate solution (1:1, 250 mL) and extracted with EtOAc (2 × 500 mL). The combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(8-oxooctyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 2, 4 g, crude) as an off-white solid, which was used in the next step without further purification.

[0555] 11H NMR (400 MHz, DMSO-d6) δ 9.66 (s, 1H), 6.97 (d, J = 7.82 Hz, 1H), 6.57 (d, J = 7.82 Hz, 1H), 5.66 - 5.70 (m, 1H), 4.35 - 4.45 (m, 1H), 3.99 - 4.06 (m, 2H), 3.21 (d, J = 6.36 Hz, 1H), 2.79 - 2.82 (m, 2H), 2.66 - 2.78 (m, 2H), 2.54 - 2.64 (m, 2H), 2.37 - 2.44 (m, 3H), 2.29 - 2.36 (m, 1H), 2.18 - 2.26 (m, 1H), 2.09 - 2.17 (m, 4H), 1.97 - 2.01 (m, 5H), 1.85 - 1.92 (m, 1H), 1.66 - 1.74 (m, 1H), 1.41 - 1.56 (m, 6H), 1.22 - 1.28 (m, 8H), 1.17 (t, J = 7.09 Hz, 3H), 0.20 - 0.25 (m, 2H). LCMS: 588.66 [M+H] + 。

[0556] Step 3: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(8-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)octyl)piperazine-1-carboxylate

[0557] At 0 °C under an argon atmosphere, (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl piperazine-1-carboxylate (Intermediate A, 4.0 g, 7.50 mmol, 1.0 equiv) in a stirred solution of MeOH (40 mL, 10 vol) was added (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(8-oxooctyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 2, 4.4 g, 7.50 mmol, 1.0 equiv) and acetic acid (1 mL, catalytic amount). The reaction mixture was stirred at room temperature for 1 h, and then NaCNBH3 (945 mg, 15.0 mmol, 2.0 equiv) was added at 0 °C under an argon atmosphere. Subsequently, the resulting reaction mixture was stirred at room temperature until TLC indicated complete consumption of the starting materials, quenched with saturated sodium bicarbonate solution (250 mL) and extracted with ethyl acetate (2 × 500 mL). The combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was purified by preparative HPLC (column: spherical-C18, 40 um, 100 A; mobile phase A: 0.1% FA / water; mobile phase B: acetonitrile; flow rate: 25 mL / min, diluent: THF + DMSO). The pure fractions were combined and lyophilized under reduced pressure to obtain (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(8-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)octyl)piperazine-1-carboxylate as an off-white solid (1.19 g, 14%).

[0558] 11H NMR (400 MHz, DMSO-d6) δ 8.92 - 8.97 (m, 1H), 8.08 - 8.15 (m, 2H), 7.61 - 7.66 (m, 1H), 7.33 - 7.35 (m, 1H), 6.96 - 7.00 (m, 2H), 6.57 - 6.61 (m, 2H), 6.49 - 6.54 (m, 1H), 5.66 - 5.69 (m, 1H), 5.42 - 5.45 (m, 2H), 5.30 - 5.33 (m, 2H), 4.38 - 4.41 (m, 1H), 3.67 - 3.78 (m, 4H), 3.44 - 3.51 (m, 2H), 3.20 - 3.27 (m, 4H), 2.82 (s, 3H), 2.39 - 2.46 (m, 3H), 2.31 - 2.37 (m, 3H), 2.19 - 2.21 (m, 7H), 2.08 - 2.10 (m, 4H), 1.98 - 2.01 (m, 4H), 1.98 - 2.02 (m, 4H), 1.83 - 1.93 (m, 3H), 1.62 - 1.78 (m, 1H), 1.42 - 1.51 (m, 5H), 1.28 (br s, 10H), 0.86 - 0.92 (m, 4H), 0.24 - 0.25 (m, 3H). LCMS: 1105.5 [M+H] + 。

[0559] Example S8: Preparation of (S)-10-((Dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(5-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)pentyl)piperazine-1-carboxylate (Compound No. 8)

[0560]

[0561] Step 1: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((5-hydroxypentyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1)

[0562] At 0 °C under an argon atmosphere, 5-bromopentan-1-ol (SM-2, 16.29 g, 97.6 mmol, 5.0 eq) and NaHCO3 (4.91 g, 58.56 mmol, 3.0 eq) were added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methylamino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (SM-1, 9.0 g, 19.52 mmol, 1.0 eq) in EtOH (90 mL, 10 vol) and H2O (45 mL, 5 vol). The resulting reaction mixture was stirred at 80 °C until TLC indicated complete consumption of the starting material. Then the reaction mixture was diluted with ice water (250 mL), extracted with EtOAc (2 × 500 mL), the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting crude product was purified by flash column (silica gel, 100 - 200 mesh) eluting with 20 - 50% EtOAc / hexane. The pure fractions were combined and concentrated under reduced pressure to afford (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((5-hydroxypentyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1, 2.2 g, 20%) as an off-white solid.

[0563] 1 H NMR (400 MHz, DMSO-d6) δ 6.98 (d, J = 8.31 Hz, 2H), 6.55 - 6.60 (m, 2H), 5.67 (s, 1H), 4.29 - 4.42 (m, 2H), 4.00 - 4.07 (m, 1H), 3.36 (q, J = 5.87 Hz, 2H), 3.19 - 3.24 (m, 2H), 2.81 (s, 3H), 2.66 - 2.77 (m, 2H), 2.54 - 2.64 (m, 2H), 2.31 - 2.40 (m, 1H), 2.19 - 2.24 (m, 1H), 2.12 - 2.18 (m, 2H), 2.07 - 2.10 (m, 3H), 1.94 - 2.02 (m, 5H), 1.86 - 1.94 (m, 1H), 1.65 - 1.75 (m, 2H), 1.39 - 1.49 (m, 5H), 1.23 - 1.33 (m, 2H), 1.17 (t, J = 7.09 Hz, 1H), 0.23 (s, 3H). LCMS: 548.4 [M+H] + .

[0564] Step 2: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(5-oxopentyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 2)

[0565] Under an argon atmosphere at 0 °C, Dess-Martin periodinane (DMP, 3.09 g, 7.3 mmol, 2.0 eq) was added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((5-hydroxypentyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1, 2 g, 3.65 mmol, 1.0 eq) in EtOAc (40 mL, 20 vol). The resulting reaction mixture was stirred at 80 °C until TLC indicated complete consumption of the starting material. Subsequently, the reaction mixture was quenched with a mixture of Na2S2O3 and saturated sodium bicarbonate solution (1:1, 250 mL), extracted with EtOAc (2 × 500 mL), and the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(5-oxopentyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 2, 2 g, crude) as an off-white solid, which was used in the next step without further purification.

[0566] 1 1H NMR (400 MHz, DMSO-d6) δ 9.64 (s, 1H), 6.98 (d, J = 8.31 Hz, 2H), 6.59 (d, J = 8.31 Hz, 2H), 5.67 (s, 1H), 4.36 - 4.43 (m, 1H), 3.98 - 4.07 (m, 1H), 3.19 - 3.27 (m, 2H), 2.81 (s, 3H), 2.53 - 2.78 (m, 5H), 2.29 - 2.49 (m, 3H), 2.12 - 2.25 (m, 3H), 2.10 (s, 3H), 1.84 - 2.05 (m, 7H), 1.59 - 1.80 (m, 2H), 1.42 - 1.56 (m, 3H), 1.17 (t, J = 7.09 Hz, 1H), 0.23 (s, 3H). LCMS: 546.68 [M+H]+ 。

[0567] Step 3: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(5-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)pentyl)piperazine-1-carboxylate

[0568] At 0 °C under an argon atmosphere, (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl piperazine-1-carboxylate (Intermediate A, 4.0 g, 7.50 mmol, 1.0 equiv) in a stirred solution of MeOH (40 mL, 10 vol) was added (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(5-oxopentyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 2, 4.1 g, 7.50 mmol, 1.0 equiv) and acetic acid (1 mL, catalytic amount). The reaction mixture was stirred at room temperature for 1 h and NaCNBH3 (945 mg, 15.0 mmol, 2.0 equiv) was added at 0 °C under an argon atmosphere. The resulting reaction mixture was then stirred at room temperature until TLC indicated complete consumption of the starting materials. The reaction mixture was then quenched with saturated sodium bicarbonate solution (250 mL) and extracted with ethyl acetate (2 × 500 mL). The combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting crude product was purified by preparative HPLC (column: spherical-C18, 40 um, 100 A; mobile phase A: 0.1% FA / water; mobile phase B: acetonitrile; flow rate: 25 mL / min, diluent: THF + DMSO). The pure fractions were combined and lyophilized under reduced pressure to obtain (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(5-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)pentyl)piperazine-1-carboxylate as an off-white solid (1.12 g, 14%).

[0569] 11H NMR (400 MHz, DMSO-d6) δ ppm 8.94 (s, 1H), 8.16 (s, 2H), 8.11 (d, J = 9.29 Hz, 1H), 7.63 (d, J = 8.80 Hz, 1H), 7.34 (s, 1H), 6.99 (d, J = 7.82 Hz, 2H), 6.60 (d, J = 7.83 Hz, 2H), 5.66 - 5.69 (m, 1H), 5.43 (s, 2H), 5.31 (br s, 2H), 4.37 - 4.42 (m, 1H), 3.76 (br s, 2H), 3.67 - 3.72 (m, 2H), 3.44 - 3.48 (m, 3H), 3.21 - 3.30 (m, 5H), 2.83 (s, 3H), 2.52 - 2.79 (m, 5H), 2.29 - 2.44 (m, 6H), 2.20 (s, 6H), 2.10 (s, 3H), 2.00 (s, 3H), 1.80 - 1.94 (m, 3H), 1.64 - 1.77 (m, 1H), 1.39 - 1.57 (m, 4H), 1.24 - 1.38 (m, 3H), 0.89 (t, J = 6.85 Hz, 3H), 0.21 - 0.27 (m, 3H). LCMS: 1063.2 [M+H] + 。

[0570] Example S9: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoyl)piperazine-1-carboxylate (Compound No. 9)

[0571]

[0572] Step 1: Preparation of 4-((2-carboxypropan-2-yl)amino)-2-fluorobenzoic acid (Intermediate 1)

[0573] At room temperature, 2-amino-2-methylpropanoic acid (SM-2, 14.1 g, 136.98 mmol, 3.0 equiv), N,N-dimethylglycine (2.35 g, 22.83 mmol, 0.5 equiv), K2CO3 (31.5 g, 228.3 mmol, 5.0 equiv), Cu powder (575 mg, 9.13 mmol, 0.2 equiv), and copper(I) iodide (1.73 g, 9.13 mmol, 0.2 equiv) were added to a stirred solution of 4-bromo-2-fluorobenzoic acid (SM-1, 10.0 g, 45.66 mmol, 1.0 equiv) in DMF (100 mL, 10 vol) and water (10 mL, 1 vol). The reaction mixture was stirred at 110 °C until TLC indicated complete consumption of the starting material. Subsequently, the reaction mixture was diluted with ice water (500 mL) and acidified to pH ~4 with 6N HCl. Then the resulting aqueous solution was extracted with ethyl acetate (2 × 1 L), and the combined organic layers were washed with brine solution (300 mL), dried over sodium sulfate, filtered, concentrated under reduced pressure, and recrystallized from DCM to give 4-((2-carboxypropan-2-yl)amino)-2-fluorobenzoic acid as an off-white solid (Intermediate 1, 6.2 g, 56%).

[0574] 1 H NMR (400 MHz, DMSO-d6) δ 12.52 (br s, 2H), 7.59 (t, J = 8.79 Hz, 1H), 6.96 (s, 1H), 6.33 (dd, J = 8.79, 1.85 Hz, 1H), 6.15 (dd, J = 14.57, 1.62 Hz, 1H), 1.44 (s, 6H). LCMS: 242.15 [M+H] + 。

[0575] Step 2: Preparation of methyl 2-fluoro-4-((1-methoxy-2-methyl-1-oxopropan-2-yl)amino)benzoate (Intermediate 2)

[0576] At room temperature, MeI (3.1 mL, 51.45 mmol, 2.0 equiv) and K2CO3 (53.1 g, 385.5 mmol, 15.0 equiv) were added to a stirred solution of 4-((2-carboxypropan-2-yl)amino)-2-fluorobenzoic acid (Intermediate 1, 6.2 g, 25.72 mmol, 1.0 equiv) in DMF (70 mL, 10 vol). The reaction mixture was stirred at ambient temperature until TLC indicated complete consumption of the starting material. Subsequently, the reaction mixture was diluted with ice water (500 mL), filtered, and dried to give methyl 2-fluoro-4-((1-methoxy-2-methyl-1-oxopropan-2-yl)amino)benzoate as an off-white solid (Intermediate 2, 4.82 g, 69%).

[0577] 1 1H NMR (400 MHz, DMSO-d6) δ 7.61 (t, J = 8.80 Hz, 1H), 7.11 (s, 1H), 6.29 (dd, J = 8.80, 2.45 Hz, 1H), 6.14 (dd, J = 14.67, 1.96 Hz, 1H), 3.74 (s, 3H), 3.63 (s, 3H), 1.48 (s, 6H). LCMS: 270.10 [M+H] + 。

[0578] Step 3: Preparation of methyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoate (Intermediate 3)

[0579] To a stirred solution of methyl 2-fluoro-4-((1-methoxy-2-methyl-1-oxopropan-2-yl)amino)benzoate (Intermediate 2, 4.8 g, 17.84 mmol, 1.0 eq) in DMSO (7.2 mL, 1.5 vol) at room temperature was added 4-isothiocyanato-2-(trifluoromethyl)benzonitrile (SM-3, 8.5 g, 37.59 mmol, 2.1 eq). The reaction mixture was stirred at 90 °C until TLC indicated complete consumption of the starting material. Subsequently, the reaction mixture was diluted with ice water (500 mL), extracted with ethyl acetate (2 × 1 L), and the combined organic layers were washed with brine solution (300 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting crude product was purified by column chromatography (silica gel, 100 - 200 mesh) eluting with 20 - 30% ethyl acetate / hexane. The pure fractions were combined and concentrated under reduced pressure to give methyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoate (Intermediate 3, 5 g, 60%) as an off-white solid.

[0580] 1 1H NMR (400 MHz, DMSO-d6) δ 8.41 (d, J = 8.31 Hz, 1H), 8.29 (d, J = 1.47 Hz, 1H), 8.06 - 8.11 (m, 2H), 7.51 (dd, J = 11.25, 1.96 Hz, 1H), 7.41 (dd, J = 8.31, 1.96 Hz, 1H), 3.90 (s, 3H), 1.55 (s, 6H). LCMS: 466.50 [M+H] + 。

[0581] Step 4: Preparation of 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoic acid (Intermediate 4)

[0582] Under ambient temperature and argon atmosphere, to a flask containing methyl 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoate (Intermediate 3, 3 g, 6.45 mmol, 1.0 equiv) in MeOH:THF:H2O (1:1:1, 30 mL, 10 vol) was added LiOH (810 mg, 19.35 mmol, 3.0 equiv). The reaction mixture was stirred at ambient temperature until TLC indicated complete consumption of the starting material. The reaction mixture was concentrated under reduced pressure, diluted with water (10 mL), acidified to pH ~ 3 with citric acid, the resulting solid was filtered and dried to give 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoic acid (Intermediate 4, 2.6 g, 89%) as an off-white solid.

[0583] 1 1H NMR (400 MHz, DMSO-d6) δ 8.40 (d, J = 8.31 Hz, 1H), 8.29 (s, 1H), 8.08 (d, J = 8.31 Hz, 1H), 7.94 (t, J = 8.07 Hz, 1H), 7.36 (d, J = 10.76 Hz, 1H), 7.29 (d, J = 8.31 Hz, 1H), 1.54 (s, 6H). LCMS: 452.20 [M+H] + 。

[0584] Step 5: Preparation of ethyl 6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoate (Intermediate 5)

[0585] At room temperature, ethyl 6-aminohexanoate hydrochloride (SM-4, 1.68 g, 8.64 mmol, 1.5 eq), EDC·HCl (1.65 g, 8.64 mmol, 1.5 eq), HOBt (1.67 g, 8.64 mmol, 1.5 eq), DMAP (70 mg, 0.57 mmol, 0.1 eq) and DIPEA (3.18 mL, 17.28 mmol, 3.0 eq) were added to a stirred solution of 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzoic acid (Intermediate 4, 2.6 g, 5.76 mmol, 1.0 eq) in DMF (26 mL, 10 vol). The reaction mixture was stirred at ambient temperature until TLC indicated complete consumption of the starting material, diluted with ice water (100 mL) and extracted with ethyl acetate (2 × 250 mL). The combined organic layers were washed with brine solution (200 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. Purification was carried out by flash column chromatography (silica gel, 100 - 200 mesh) eluting with 40 - 60% ethyl acetate / hexane. The pure fractions were combined and concentrated under reduced pressure to give ethyl 6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoate (Intermediate 5, 2.39 g, 70%) as an off-white solid.

[0586] 1 H NMR (400 MHz, DMSO-d6) δ 8.48 (t, J = 5.38 Hz, 1H), 8.39 (d, J = 8.31 Hz, 1H), 8.28 (d, J = 1.22 Hz, 1H), 8.07 (dd, J = 8.31, 1.47 Hz, 1H), 7.74 (t, J = 8.07 Hz, 1H), 7.41 (dd, J = 10.64, 1.59 Hz, 1H), 7.31 (dd, J = 8.07, 1.71 Hz, 1H), 4.04 (q, J = 7.09 Hz, 2H), 3.24 (q, J = 6.60 Hz, 2H), 2.28 (t, J = 7.34 Hz, 2H), 1.55 - 1.59 (m, 2H), 1.53 (s, 6H), 1.47 - 1.52 (m, 2H), 1.28 - 1.38 (m, 2H), 1.16 (t, J = 7.09 Hz, 3H). LCMS: 593.2 [M+H] + 。

[0587] Step 6: Preparation of 6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoic acid (Intermediate 6)

[0588] Under ambient temperature and argon atmosphere, LiOH (509 mg, 12.15 mmol, 3.0 eq) was added to a flask containing ethyl 6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoate (Intermediate 5, 2.4 g, 4.05 mmol, 1.0 eq) in EtOH:THF:H2O (1:1:1, 24 mL, 10 vol). The reaction mixture was stirred at ambient temperature until TLC indicated complete consumption of the starting material. The reaction mixture was concentrated under reduced pressure, diluted with water (10 mL), acidified to pH ~ 3 with citric acid, and the resulting solid was filtered and dried to give 6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoic acid as an off-white solid (Intermediate 6, 2.0 g, 87%).

[0589] 1 H NMR (400 MHz, DMSO-d6) δ 11.99 (s, 1H), 8.49 (t, J = 5.38 Hz, 1H), 8.40 (d, J = 8.31 Hz, 1H), 8.29 (s, 1H), 8.08 (d, J = 8.31 Hz, 1H), 7.75 (t, J = 8.07 Hz, 1H), 7.42 (d, J = 10.27 Hz, 1H), 7.33 (dd, J = 8.07, 1.22 Hz, 1H), 3.26 (q, J = 6.36 Hz, 2H), 2.22 (t, J = 7.34 Hz, 2H), 1.47 - 1.59 (m, 10H), 1.29 - 1.39 (m, 2H). LCMS: 565.10 [M+H] + 。

[0590] Step 7: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoyl)piperazine-1-carboxylate

[0591] Under ambient temperature and argon atmosphere, piperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (Intermediate A, 945 mg, 1.77 mmol, 1.0 equiv), HATU (1.0 g, 2.65 mmol, 1.5 equiv) and DIPEA (0.97 mL, 3.98 mmol, 3.0 equiv) were added to a flask containing 6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoic acid (Intermediate 6, 1.0 g, 1.77 mmol, 1.0 equiv) in DMF (10 mL). The reaction mixture was stirred at ambient temperature until TLC indicated complete consumption of the starting materials, diluted with ice water (100 mL) and extracted with ethyl acetate (2 × 200 mL). The combined organic layers were washed with brine solution (200 mL), dried over sodium sulfate, filtered, concentrated under reduced pressure and purified by flash column chromatography (silica gel, 100 - 200 mesh) eluting with 0 - 5% MeOH / DCM. The pure fractions were combined and concentrated under reduced pressure to give 4-(6-(4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl)-2-fluorobenzamido)hexanoyl)piperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (605 mg, 31%) as an off-white solid.

[0592] 11H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.47 - 8.54 (m, 1H), 8.40 (d, J = 8.38 Hz, 1H), 8.29 (s, 1H), 8.04 - 8.16 (m, 2H), 7.76 (t, J = 7.63 Hz, 1H), 7.66 (d, J = 8.63 Hz, 1H), 7.43 (d, J = 10.76 Hz, 1H), 7.30 - 7.38 (m, 2H), 6.51 (s, 1H), 5.40 - 5.46 (m, 2H), 5.28 - 5.35 (m, 2H), 3.41 - 3.85 (m, 10H), 2.35 - 2.44 (m, 2H), 2.20 (s, 6H), 1.80 - 1.95 (m, 2H), 1.50 - 1.64 (m, 10H), 1.33 - 1.44 (m, 2H), 1.20 - 1.30 (m, 2H), 0.89 (t, J = 6.69 Hz, 3H). LCMS: 1080.30 [M+H] + . HPLC purity 96.1%.

[0593] Example S10: Preparation of (S)-10-((4-(1-(6-(((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-carbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-methylpiperazine-1-carboxylate (Compound No. 10)

[0594]

[0595] Step 1: Preparation of (S)-10-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-methylpiperazine-1-carboxylate (Intermediate A5)

[0596] At room temperature, to a stirred solution of tert-butyl (S)-4-((4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazine-1-carboxylate (Intermediate 3, 300 mg, 0.53 mmol, 1.0 equiv) in DCM (15 mL) was added DIPEA (0.3 mL, 1.60 mmol, 3 equiv) and DMAP (20 mg, 0.16 mmol, 0.25 equiv). Subsequently, 4-methylpiperazine-1-carbonyl chloride (SM-1, 129 mg, 0.80 mmol, 1.5 equiv) in DCM (5 mL) was added and the mixture was stirred for 16 h. The reaction progress was monitored by TLC. After completion of the reaction, the reaction mixture was washed with water (100 mL) and extracted with DCM (2 × 200 mL). The combined organic extracts were washed with brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash column eluting with 6% methanol / DCM to give Intermediate A5 (260 mg, 70%) as an off-white solid.

[0597] 1 H NMR (400 MHz, DMSO-d6) δ 9.01 (s, 1H), 8.12 (d, J = 9.29 Hz, 1H), 7.63 (d, J = 8.80 Hz, 1H), 7.34 (s, 1H), 6.51 (s, 1H), 5.43 (s, 2H), 5.30 (s, 2H), 3.87 (s, 2H), 3.68 - 3.70 (m, 2H), 3.48 - 3.50 (m, 2H), 3.20 - 3.25 (m, 2H), 2.38 - 2.42 (m, 8H), 2.25 (s, 3H), 1.84 - 1.91 (m, 2H), 1.39 (s, 9H), 1.25 (br dd, J = 11.98, 6.11 Hz, 2H), 0.89 (br t, J = 7.09 Hz, 3H). LCMS: 689.4 [M + H] + 。

[0598] Step 2: Preparation of (S)-4-ethyl-4-hydroxy-3,14-dioxo-10-(piperazin-1-ylmethyl)-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl 4-methylpiperazine-1-carboxylate trifluoroacetate (Intermediate A6)

[0599] Under a nitrogen atmosphere at 0 °C, trifluoroacetic acid (TFA, 0.3 mL, 3.5 mmol, 10 eq) was added to a stirred solution of (S)-10-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-methylpiperazine-1-carboxylate (Intermediate A6, 250 mg, 0.36 mmol, 1.0 eq) in DCM (10 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. After completion of the reaction, the solvent was evaporated under reduced pressure, washed with diethyl ether (20 mL), and dried in vacuo to give Intermediate A6 as an off-white solid (210 mg, 98%).

[0600] 1 H NMR (400 MHz, DMSO-d6) δ 9.04 (s, 1H), 8.66 (br s, 2H), 8.18 (d, J = 9.29 Hz, 1H), 7.69 (d, J = 9.29 Hz, 1H), 7.35 (s, 1H), 6.55 (br s, 1H), 5.44 (s, 2H), 5.31 (s, 2H), 4.40 (brs, 1H), 4.16 (br s, 1H), 3.96 - 3.99 (m, 3H), 3.49 - 3.54 (m, 3H), 3.20 - 3.22 (m, 2H), 3.01 - 3.05 (m, 4H), 2.90 (s, 3H), 2.62 - 2.68 (m, 4H), 1.82 - 1.93 (m, 2H), 0.89 (br t, J = 7.09 Hz, 3H). LCMS: 589.2 [M+H] + 。

[0601] Step 3: Preparation of (S)-10-((4-(1-(6-(((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-methylpiperazine-1-carboxylate

[0602] At room temperature, to a stirred solution of (S)-4-ethyl-4-hydroxy-3,14-dioxo-10-(piperazin-1-ylmethyl)-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-methylpiperazine-1-carboxylate trifluoroacetate (Intermediate A6, 200 mg, 0.34 mmol, 1.0 equiv) and 1-(6-(((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carboxylic acid (Intermediate 13, 185 mg, 0.37 mmol, 1.1 equiv) in DMF (5 mL) was added HATU (243 mg, 0.68 mmol, 2 equiv) and DIPEA (0.177 mL, 1.02 mmol, 3 equiv), and the mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was washed with water (60 mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic extracts were washed with brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash column eluting with 5% methanol / DCM to give the title compound as an off-white solid (86 mg, 24%).

[0603] 1 H NMR (400 MHz, DMSO-d6) δ 9.05 (s, 1H), 8.57 (br d, J = 8.25 Hz, 1H), 8.14 (d, J = 9.26 Hz, 1H), 7.83 (dd, J = 15.76, 9.13 Hz, 2H), 7.65 (d, J = 9.26 Hz, 1H), 7.30 - 7.45 (m, 3H), 7.13 (dd, J = 8.82, 2.44 Hz, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.32 (s, 2H), 4.40 - 4.58 (m, 3H), 3.84 - 3.94 (m, 3H), 3.69 - 3.71 (m, 2H), 3.37 - 3.56 (m, 6H), 3.11 (br t, J = 11.94 Hz, 3H), 2.38 - 2.42 (m, 6H), 2.26 (s, 3H), 2.05

[0604] - 2.14 (m, 2H), 1.84 - 1.95 (m, 4H), 1.46 - 1.76 (m, 10H), 0.89 (t, J = 7.32 Hz, 3H). LCMS: 1055.65 [M + H] + 。HPLC purity 96.0%.

[0605] Example S11: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl 4-(1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)-cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)piperazine-1-carboxylate (Compound No. 11)

[0606]

[0607] Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate A)

[0608]

[0609] Step A1: Preparation of tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate (Intermediate A1)

[0610] To a stirred solution of tert-butyl piperazine-1-carboxylate (SM-1, 5 g, 26.8 mmol, 1.0 equiv) in DCM (100 mL) at 0 °C was added pyridine (2.97 g, 37.6 mmol, 1.4 equiv) and triphosgene (3.19 g, 10.7 mmol, 0.4 equiv). The reaction mixture was warmed to room temperature and stirred for 2 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was washed with water (100 mL) and extracted with DCM (2 × 100 mL). The combined organic extracts were washed again with water (200 mL), brine (200 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give Intermediate A1 as a crude oil (6.0 g, 90%).

[0611] 1 H NMR (400 MHz, DMSO-d6) δ 3.64 (d, J = 4.40 Hz, 2H), 3.52 (br s, 2H), 3.27 - 3.47 (m, 3H), 2.93 - 3.21 (m, 1H), 1.41 (s, 9H).

[0612] Step A2: Preparation of (S)-1-(tert-butyl) 4-(10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl)piperazine-1,4-dicarboxylate (Intermediate A2)

[0613] To a stirred solution of (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-2, 10 g, 23.7 mmol, 1.0 equiv) in DCM (250 mL) at 0 °C was added DIPEA (15.3 g, 118 mmol, 5 equiv) and DMAP (724 mg, 5.9 mmol, 0.25 equiv), followed by the dropwise addition of tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate (Intermediate A1, 5.89 g, 23.7 mmol, 1 equiv) in DCM (100 mL) over 10 min. The resulting reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the mixture was washed with water (100 mL) and extracted with DCM (3 × 100 mL). The combined organic extracts were washed again with water (200 mL), brine (200 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude was purified by combiflash column using 7% methanol / DCM to give Intermediate A2 as an off-white solid (10 g, 66%).

[0614] 1 H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.11 (d, J = 9.29 Hz, 1H), 7.65 (d, J = 9.29 Hz, 1H), 7.34 (s, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 3.67 - 3.79 (m, 4H), 3.41 - 3.55 (m, 6H), 2.20 (s, 6H), 1.80 - 1.93 (m, 2H), 1.44 (s, 9H), 0.89 (t, J = 7.34 Hz, 3H). LCMS: 634.2 [M+H] + .

[0615] Step A3: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate A)

[0616] Under a nitrogen atmosphere at 0 °C, trifluoroacetic acid (TFA, 3 mL) was added to a stirred solution of piperazine-1,4-dicarboxylic acid (S)-1-(tert-butyl) ester · 4-(10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate A2, 1 g, 15 mmol, 1.0 equiv) in DCM (20 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, saturated NaHCO3 solution (50 mL) was added, and the aqueous solution was extracted with EtOAc (2 × 100 mL). The solvent was evaporated under reduced pressure to give Intermediate A as an off-white solid (1.02 g, crude).

[0617] 1 1H NMR (400 MHz, DMSO-d6) δ 10.12 (br s, 1H), 9.03 - 9.27 (m, 2H), 8.35 (d, J = 9.29 Hz, 1H), 7.86 (d, J = 9.29 Hz, 1H), 7.37 (s, 1H), 6.57 (br s, 1H), 5.45 (br s, 2H), 5.33 (br s, 2H), 4.86 (br s, 2H), 3.90 - 3.95 (m, 2H), 3.68 - 3.71 (m, 2H), 3.30 (d, J = 12.23 Hz, 2H), 2.89 (s, 6H), 1.80 - 1.95 (m, 3H), 1.08 (t, J = 7.09 Hz, 1H), 0.89 (br t, J = 6.85 Hz, 3H). LCMS: 534.2 [M+H] + 。

[0618] Step 1: Preparation of tert-butyl ((1r,4r)-4-((3-chloro-4-cyanophenyl)amino)cyclohexyl)carbamate (Intermediate 1)

[0619] At room temperature, ((1r,4r)-4-aminocyclohexyl)carbamic acid tert-butyl ester (SM-2, 5.5 g, 25 mmol, 1.0 equiv) and K2CO3 (7.1 g, 51 mmol, 2 equiv) were added to a stirred solution of 2-chloro-4-fluorobenzonitrile (SM-1, 4 g, 25 mmol, 1.0 equiv) in DMSO (40 mL). The reaction mixture was heated to 90 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with ice water (200 mL) and extracted with ethyl acetate (2 × 400 mL). The combined organic extracts were washed with water (200 mL), brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by a combiflash column eluting with 64% ethyl acetate / heptane to give Intermediate 1 (7.1 g, 78%) as an off-white solid.

[0620] 1 1H NMR (400 MHz, DMSO-d6) δ 7.48 (d, J = 8.80 Hz, 1H), 6.86 (d, J = 6.85 Hz, 1H), 6.80 (d, J = 6.85 Hz, 1H), 6.75 (br s, 1H), 6.59 (d, J = 8.80 Hz, 1H), 3.20 - 3.23 (m, 2H), 1.91 (d, J = 11.25 Hz, 2H), 1.79 (d, J = 10.76 Hz, 2H), 1.38 (s, 9H), 1.13 - 1.33 (m, 4H).

[0621] Step 2: Preparation of ((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamic acid tert-butyl ester (Intermediate 2)

[0622] Under a nitrogen atmosphere at 0 °C, NaH (63%, 500 mg, 21 mmol, 1.3 equiv) was added portionwise to a stirred solution of ((1r,4r)-4-((3-chloro-4-cyanophenyl)amino)cyclohexyl)carbamic acid tert-butyl ester (Intermediate 1, 5.5 g, 15 mmol, 1.0 equiv) in DMF (25 mL). The reaction mixture was warmed to room temperature and stirred for 30 min, then methyl iodide (1.1 mL, 21 mmol, 1.3 equiv) was added dropwise at 0 °C. The reaction mixture was warmed to room temperature and stirred for 3 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (150 mL) was added and the mixture was extracted with ethyl acetate (2 × 200 mL). The solvent was evaporated under reduced pressure to give the crude product, which was purified by a combiflash column eluting with 15% ethyl acetate / heptane to give Intermediate 2 (3.2 g, 55%) as an off-white solid.

[0623] LCMS: 364.2 [M+H] + 。

[0624] Step 3: Preparation of 4-(((1R,4r)-4-aminocyclohexyl)(methyl)amino)-2-chlorobenzonitrile trifluoroacetate (Intermediate 3)

[0625] Under a nitrogen atmosphere at 0 °C, TFA (10 mL, 10 vol) was added to a stirred solution of tert-butyl ((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamate Intermediate 2 (1.1 g, 3 mmol, 1.0 equiv) in DCM (10 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, washed with diethyl ether (20 mL), and dried in vacuo to give Intermediate 3 (852 mg, 74%) as an off-white solid.

[0626] LCMS: 264.1 [M+H] + 。

[0627] Step 4: Preparation of 6-chloro-N-((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)pyridazine-3-carboxamide (Intermediate 4)

[0628] At room temperature, HATU (1.21 g, 12 mmol, 1.5 equiv) and DIPEA (0.74 mL, 16 mmol, 2 equiv) were added to a stirred solution of 4-(((1r,4r)-4-aminocyclohexyl)(methyl)amino)-2-chlorobenzonitrile trifluoroacetate (Intermediate 3, 800 mg, 8 mmol, 1.0 equiv) and 6-chloropyridazine-3-carboxylic acid (SM-3, 336 mg, 8 mmol, 1.0 equiv) in DMF (3 mL), and the resulting reaction mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (50 mL) was added and the mixture was extracted with ethyl acetate (2 × 100 mL). The combined organic extracts were washed with water (100 mL), brine (100 mL), and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to give a crude product, which was purified by a combiflash column eluting with 4% methanol / DCM to give Intermediate 4 (700 mg, 81%) as an off-white solid.

[0629] 11H NMR (400 MHz, DMSO-d6) δ 8.44 (d, J = 9.25 Hz, 1H), 8.21 (d, J = 9.25 Hz, 1H), 7.95 (s, 1H), 7.68 (dd, J = 8.32, 4.62 Hz, 1H), 7.58 (d, J = 9.25 Hz, 1H), 6.80 (dd, J = 9.25, 2.31 Hz, 1H), 3.79 - 3.89 (m, 1H), 3.70 - 3.73 (m, 1H), 2.69 (s, 3H), 1.84 - 1.87 (m, 2H), 1.51 - 1.77 (m, 6H). LCMS: 404.1 [M+H] + 。

[0630] Step 5: Preparation of Ethyl 1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamoyl)-pyridazin-3-yl)piperidine-4-carboxylate (Intermediate 5)

[0631] To a stirred solution of 6-chloro-N-((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)-pyridazine-3-carboxamide (Intermediate 4, 250 mg, 0.6 mmol, 1.0 equiv) and ethyl piperidine-4-carboxylate (SM-4, 0.1 mL, 0.6 mmol, 1.0 equiv) in DMF (2 mL) at room temperature was added K2CO3 (129 mg, 0.9 mmol, 1.5 equiv). The reaction mixture was heated to 80 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (50 mL) was added and the aqueous mixture was extracted with ethyl acetate (2 × 100 mL). The combined organic extracts were washed with water (100 mL), brine (100 mL) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to give the crude product, which was purified by combiflash column eluting with 62% ethyl acetate / heptane to give Intermediate 5 as a pale yellow solid (198 mg, 60%).

[0632] 11H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.82 (d, J = 9.78 Hz, 1H), 7.60 (d, J = 9.29 Hz, 1H), 7.36 (d, J = 9.29 Hz, 1H), 6.94 (d, J = 2.45 Hz, 1H), 6.83 (dd, J = 9.29, 2.45 Hz, 1H), 4.35 - 4.37 (m, 2H), 4.08 (q, J = 7.34 Hz, 2H), 3.76 - 3.88 (m, 2H), 3.12 - 3.22 (m, 2H), 2.85 (s, 3H), 2.69 - 2.76 (m, 4H), 1.89 - 1.95 (m, 3H), 1.64 - 1.76 (m, 4H), 1.54 - 1.60 (m, 2H), 1.19 (t, J = 7.09 Hz, 3H). LCMS: 525.2 [M+H] + 。

[0633] Step 6: Preparation of 1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamoyl)-pyridazin-3-yl)piperidine-4-carboxylic acid (Intermediate 6)

[0634] At room temperature, LiOH (27 mg, 1.1 mmol, 3 eq) was added to a stirred solution of ethyl 1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)-carbamoyl)pyridazin-3-yl)piperidine-4-carboxylate (Intermediate 5, 190 mg, 0.36 mmol, 1.0 eq) in THF (2 mL) and water (1 mL), and the mixture was stirred for 5 h. The reaction progress was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, water (10 mL) was added, and the reaction mixture was acidified to pH 6 with 1 M HCl (6 mL). The precipitate was filtered off and dried in vacuo to give Intermediate 6 (158 mg, 87%) as a pale yellow solid.

[0635] 11H NMR (400 MHz, DMSO-d6) δ 12.26 (br s, 1H), 8.50 (d, J = 8.31 Hz, 1H), 7.82 (d, J = 9.29 Hz, 1H), 7.60 (d, J = 8.80 Hz, 1H), 7.36 (d, J = 9.78 Hz, 1H), 6.94 (br s, 1H), 6.83 (d, J = 8.80 Hz, 1H), 4.34 - 4.36 (m, 2H), 3.73 - 3.93 (m, 1H), 3.16 (t, J = 11.74 Hz, 2H), 2.85 (s, 3H), 2.61 (t, J = 10.27 Hz, 1H), 1.92 (d, J = 10.76 Hz, 4H), 1.48 - 1.80 (m, 8H), 1.14 - 1.28 (m, 1H). LCMS: 497.2 [M+H] + 。

[0636] Step 7: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl 4-(1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)piperazine-1-carboxylate

[0637] To a stirred solution of 1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carboxylic acid (Intermediate 6, 110 mg, 0.22 mmol, 1.0 equiv) and (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate A, 118 mg, 0.22 mmol, 1.0 equiv) in DMF (3 mL) at room temperature was added HATU (126 mg, 0.33 mmol, 1.5 equiv) and DIPEA (0.12 mL, 0.66 mmol, 3 equiv). The mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (25 mL) was added and the aqueous mixture was extracted with ethyl acetate (2 × 30 mL). The combined organic extracts were washed with water (50 mL), brine (50 mL) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to obtain the crude product, which was purified by combiflash column chromatography eluting with 10% methanol / DCM to give the title compound as an off-white solid (65 mg, 19%).

[0638] 1 1H NMR (400 MHz, DMSO-d6) δ 8.96 (s, 1H), 8.48 (d, J = 7.82 Hz, 1H), 8.13 (d, J = 9.29 Hz, 1H), 7.84 (d, J = 9.78 Hz, 1H), 7.67 (d, J = 8.80 Hz, 1H), 7.60 (d, J = 9.29 Hz, 1H), 7.38 (d, J = 9.29 Hz, 1H), 7.34 (s, 1H), 6.93 - 6.96 (m, 1H), 6.80 - 6.86 (m, 1H), 6.51 (s, 1H), 5.43 (s, 2H), 5.32 (s, 2H), 4.47 - 4.57 (m, 2H), 3.82 - 3.90 (m, 1H), 3.78 (s, 3H), 3.43 - 3.72 (m, 6H), 3.04 - 3.21 (m, 4H), 2.85 (s, 3H), 2.22 (s, 6H), 1.85 - 1.97 (m, 5H), 1.72 - 1.84 (m, 3H), 1.54 - 1.71 (m, 7H), 0.89 (t, J = 7.09 Hz, 3H). LCMS: 1010.60 [M-H] - 。The HPLC purity is 95.8%.

[0639] Example S12: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-(heptanoyloxy)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylate (Compound No. 12)

[0640]

[0641] Step 1-1: Preparation of tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate

[0642] At 0 °C, pyridine (2.97 g, 37.6 mmol, 1.4 eq) and triphosgene (3.19 g, 10.7 mmol, 0.4 eq) were added to a stirred solution of tert-butyl piperazine-1-carboxylate (SM-2, 5 g, 26.8 mmol, 1.0 eq) in DCM (100 mL). The reaction mixture was warmed to room temperature and stirred for 2 h. The reaction progress was monitored by TLC. After completion of the starting material, the reaction mixture was washed with water (100 mL) and extracted with DCM (2 × 100 mL). The combined organic extracts were washed again with water (200 mL), brine (200 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate as a crude oil (6.0 g, 90%).

[0643] 1 H NMR (400 MHz, DMSO-d6) δ 3.64 (br d, J = 4.40 Hz, 2H), 3.52 (br s, 2H), 3.27 - 3.47 (m, 3H), 2.93 - 3.21 (m, 1H), 1.41 (s, 9H).

[0644] Step 1-2: Preparation of (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate · 4-(10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 1)

[0645] At 0 °C, DIPEA (15.3 g, 118 mmol, 5 equiv) and DMAP (724 mg, 5.9 mmol, 0.25 equiv) were added to a stirred solution of (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-1, 10 g, 23.7 mmol, 1.0 equiv) in DCM (100 mL, 10 vol). Subsequently, tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate (5.89 g, 23.7 mmol, 1 equiv) in DCM (100 mL, 10 vol) was added dropwise over 10 min. The resulting reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the mixture was washed with water (500 mL), extracted with DCM (3 × 100 mL), and the combined organic extracts were washed again with water (200 mL) and brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The obtained crude product was purified by combiflash column eluting with 7% methanol / DCM to give Intermediate 1 (10 g, 66%) as an off-white solid.

[0646] 1 H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.11 (d, J = 9.29 Hz, 1H), 7.65 (d, J = 9.29 Hz, 1H), 7.34 (s, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 3.67 - 3.79 (m, 4H), 3.41 - 3.55 (m, 6H), 2.20 (s, 6H), 1.80 - 1.93 (m, 2H), 1.44 (s, 9H), 0.89 (br t, J = 7.34 Hz, 3H). LCMS: 634.2 [M+H] + 。

[0647] Step 2: Preparation of (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate · 4-(10-((dimethylamino)methyl)-4-ethyl-4-(heptyloxy)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2)

[0648] At 0 °C under an argon atmosphere, heptanoic acid (SM-3, 2.15 g, 16.57 mmol, 1.5 eq), EDC·HCl (3.168 g, 16.57 mmol, 1.5 eq) and DMAP (606 mg, 4.97 mmol, 0.45 eq) were added to a stirred solution of (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate·4-(10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2, 7.0 g, 11.05 mmol, 1.0 eq) in DCM (70 mL, 10.0 vol). The reaction mixture was stirred at room temperature until TLC indicated complete consumption of the starting material. The reaction mixture was then diluted with ice water (250 mL), extracted with DCM (2 × 250 mL), the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product obtained was purified by flash column and the pure fractions were concentrated under reduced pressure to give (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate·4-(10-((dimethylamino)methyl)-4-ethyl-4-(heptanoyloxy)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester as an off-white solid (Intermediate 2, 6 g, 73%).

[0649] 1 H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.07 (d, J = 8.80 Hz, 1H), 7.65 (d, J = 9.29 Hz, 1H), 7.01 - 7.04 (m, 1H), 5.46 - 5.50 (m, 2H), 5.30 - 5.34 (m, 2H), 3.68 - 3.78 (m, 4H), 3.44 - 3.53 (m, 6H), 3.24 - 3.30 (m, 1H), 3.07 - 3.15 (m, 1H), 2.45 - 2.49 (m, 1H), 2.20 (s, 7H), 1.53 - 1.61 (m, 2H), 1.42 - 1.48 (m, 11H), 1.20 - 1.32 (m, 4H), 0.92 (t, J = 7.09 Hz, 3, 0.74 - 0.79 (m, 3H). LCMS: 746.5 [M+H] + 。

[0650] Step 3: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-(heptyloxycarbonyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate 3)

[0651] To a stirred solution of (S)-1-(tert-butyl) 4-(10-((dimethylamino)methyl)-4-ethyl-4-(heptyloxycarbonyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) piperazine-1,4-dicarboxylate (Intermediate 3, 5 g, 6.71 mmol, 1.0 equiv) in DCM (50 mL, 10 vol) at ambient temperature was added TFA (5.1 mL, 67.1 mmol, 10.0 equiv). The reaction mixture was stirred at room temperature until TLC indicated complete consumption of the starting material. The reaction mixture was then evaporated under reduced pressure, diluted with DCM (500 mL) and washed with saturated bicarbonate solution (2 × 250 mL), brine (250 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give (S)-10-((dimethylamino)methyl)-4-ethyl-4-(heptyloxycarbonyl)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate 3, 3 g, 69%) as a brown solid.

[0652] 1 H NMR (400 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.04 - 8.10 (m, 1H), 7.61 - 7.67 (m, 1H), 7.00 - 7.06 (m, 1H), 5.45 - 5.51 (m, 2H), 5.28 - 5.35 (m, 2H), 3.56 - 3.81 (m, 4H), 3.36 - 3.46 (m, 2H), 2.73 - 2.89 (m, 4H), 2.52 - 2.57 (m, 1H), 2.44 - 2.48 (m, 1H), 2.14 - 2.21 (m, 8H), 1.51 - 1.61 (m, 2H), 1.17 - 1.34 (m, 7H), 0.89 - 0.95 (m, 3H), 0.74 - 0.79 (m, 3H). LCMS: 646.5 [M+H] + 。

[0653] Step 4: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-(heptanoyloxy)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylate

[0654] At 0 °C under an argon atmosphere, acetic acid (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(6-oxohexyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl ester (Intermediate C, 2.59 g, 4.65 mmol) and acetic acid (1 mL, catalytic amount) were added to a stirred solution of (S)-10-((dimethylamino)methyl)-4-ethyl-4-(heptyloxy)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate 3, 3.0 g, 4.65 mmol, 1.0 equiv) in MeOH (30 mL, 10 vol). The reaction mixture was stirred at room temperature for 1 h, and NaCNBH3 (586 mg, 9.3 mmol) was added at 0 °C under an argon atmosphere. The resulting reaction mixture was stirred at room temperature until TLC showed complete consumption of the starting materials. Subsequently, the reaction mixture was quenched with saturated sodium bicarbonate solution (250 mL), extracted with ethyl acetate (2 × 500 mL), the combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was purified by preparative HPLC (column: Spherical-C18, 40 um, 100 A; mobile phase A: 0.1% FA / water; mobile phase B: acetonitrile; flow rate: 25 mL / min, diluent: THF + DMSO). The pure fractions were combined and lyophilized under reduced pressure to give (S)-10-((dimethylamino)methyl)-4-ethyl-4-(heptyloxy)-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylate as an off-white solid (1.51 g, 27%).

[0655] 1¹H NMR (400 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.07 (d, J = 9.26 Hz, 1H), 7.63 (d, J = 9.26 Hz, 1H), 6.96 - 7.04 (m, 3H), 6.56 - 6.62 (m, 2H), 5.67 (s, 1H), 5.48 (d, J = 1.25 Hz, 2H), 5.32 (d, J = 3.00 Hz, 2H), 4.40 (d, J = 6.88 Hz, 1H), 3.67 - 3.79 (m, 4H), 3.44 - 3.49 (m, 2H), 3.25 (d, J = 7.25 Hz, 3H), 2.64 - 2.83 (m, 6H), 2.53 - 2.60 (m, 3H), 2.31 - 2.48 (m, 6H), 2.08 - 2.22 (m, 15H), 1.86 - 2.01 (m, 5H), 1.65 - 1.74 (m, 2H), 1.41 - 1.62 (m, 7H), 1.18 - 1.36 (m, 12H), 0.92 (t, J = 7.44 Hz, 3H), 0.74 - 0.79 (m, 3H), 0.24 (s, 3H). LCMS: 1189.1 [M+H] + 。

[0656] Example S13: Preparation of (S)-10-((4-(1-(6-(((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl [1,4'-bipiperidine]-1'-carboxylate (Compound No. 13)

[0657]

[0658] Step 1: Preparation of (S)-10-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl [1,4'-bipiperidine]-1'-carboxylate (Intermediate 2)

[0659] At 0 °C, triethylamine (0.14 mL, 1.06 mmol, 3 eq) and triphosgene (105 mg, 0.35 mmol, 1 eq) were added to a stirred solution of SM-1 (60 mg, 0.35 mmol, 1.0 equiv) in DCM (10 mL). The reaction mixture was warmed to room temperature and stirred for 1 h. After 1 h, intermediate 1 (200 mg, 0.35 mmol, 1 eq), K2CO3 (49 mg, 0.35 mmol, 1 eq) and a catalytic amount of DMAP (20 mg) were added to the reaction mixture, and stirring was continued for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the mixture was diluted with water (100 mL) and extracted with DCM (2 × 200 mL). The combined organic extracts were washed with water (100 mL), brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash column using 4% methanol / DCM to give intermediate 2 (200 mg, 74%) as an off-white solid.

[0660] 1 H NMR (400 MHz, DMSO-d6) δ 9.01 (s, 1H), 8.11 (br d, J = 9.29 Hz, 1H), 7.63 (br d, J = 9.29 Hz, 1H), 7.34 (s, 1H), 6.51 (br s, 1H), 5.43 (br s, 2H), 5.30 (br s, 2H), 4.26 - 4.28 (m, 1H), 4.06 - 4.08 (m, 3H), 3.86 (br s, 2H), 3.25 - 3.28 (m, 6H), 3.16 - 3.18 (m, 4H), 2.85 - 2.99 (m, 1H), 2.39 - 2.43 (m, 4H), 1.80 - 1.89 (m, 6H), 1.35 - 1.46 (m, 4H), 1.32 (s, 9H), 0.89 (brt, J = 7.34 Hz, 3H). LCMS: 757.4 [M+H] + 。

[0661] Step 2: Preparation of (S)-4-ethyl-4-hydroxy-3,14-dioxo-10-(piperazin-1-ylmethyl)-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl [1,4'-bipiperidine]-1'-carboxylate (Intermediate 3)

[0662] Under a nitrogen atmosphere, at 0 °C, TFA (0.2 mL, 2.64 mmol, 10 eq) was added to a stirred solution of intermediate 2 (200 mg, 0.26 mmol, 1.0 eq) in DCM (10 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, washed with diethyl ether (25 mL) and pentane (20 mL), and dried in vacuo to give intermediate 3 (170 mg, 98%) as an off-white solid.

[0663] 1 H NMR (400 MHz, DMSO-d6) δ 9.73 (br d, J = 8.80 Hz, 1H), 9.03 (s, 1H), 8.82 (br s, 2H), 8.16 (br d, J = 9.29 Hz, 1H), 7.66 (br d, J = 9.29 Hz, 1H), 7.34 (s, 1H), 5.44 (s, 2H), 5.31 (s, 2H), 4.39 - 4.42 (m, 1H), 4.13 - 4.28 (m, 1H), 3.96 (br s, 2H), 3.47 (br d, J = 10.76 Hz, 2H), 2.84 - 3.27 (m, 8H), 2.67 - 2.71 (m, 4H), 2.14 - 2.18 (m, 2H), 1.61 - 2.08 (m, 10H), 1.44 (br d, J = 12.72 Hz, 1H), 0.89 (t, J = 7.34 Hz, 3H). LCMS: 657.20 [M+H] + 。

[0664] Step 3: Preparation of (S)-10-((4-(1-(6-(((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)piperazin-1-yl)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl [1,4'-bipiperidine]-1'-carboxylate

[0665] At room temperature, HATU (185 mg, 0.51 mmol, 2.0 equiv) and DIPEA (0.13 mL, 0.77 mmol, 3 equiv) were added to a stirred solution of intermediate 3 (170 mg, 0.25 mmol, 1.0 equiv) and intermediate 13 (125 mg, 0.25 mmol, 1.0 equiv) in DMF (5 mL), and the reaction mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic extracts were washed with water (100 mL), brine (100 mL) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to give the crude product. The crude product was purified by combiflash column eluting with 13% methanol / DCM to give the title compound as an off-white solid (108 mg, 37%).

[0666] 1 1H NMR (400 MHz, DMSO-d6) δ 9.04 (s, 1H), 8.57 (br d, J = 8.13 Hz, 1H), 8.43 (s, 1H), 8.13 (d, J = 9.13 Hz, 1H), 7.81 - 7.89 (m, 1H), 7.64 (d, J = 9.13 Hz, 1H), 7.30 - 7.40 (m, 3H), 7.13 (dd, J = 8.82, 2.31 Hz, 1H), 6.55 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 4.42 - 4.59 (m, 3H), 4.24 - 4.36 (m, 1H), 4.02 - 4.13 (m, 1H), 3.81 - 3.93 (m, 3H), 3.50 - 3.52 (m, 2H), 3.10 - 3.13 (m, 3H), 2.87 - 3.05 (m, 3H), 2.54 - 2.57 (m, 3H), 2.46 - 2.50 (m, 4H), 2.40 - 2.42 (m, 3H), 2.08 - 2.11 (m, 2H), 1.80 - 1.93 (m, 6H), 1.42 - 1.74 (m, 14H), 1.34 - 1.37 (m, 2H), 0.89 (br t, J = 7.32 Hz, 3H). LCMS: 1122.4 [M + H] + . HPLC purity 92.4%.

[0667] Example S14: Preparation of (S)-10-((dimethylamino)methyl)-4-((dimethylglycyl)oxy)-4-ethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylate (Compound 14)

[0668]

[0669] Step 1-1: Preparation of tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate

[0670] To a stirred solution of tert-butyl piperazine-1-carboxylate (SM-2, 5 g, 26.8 mmol, 1.0 equiv) in DCM (100 mL) at 0 °C was added pyridine (2.97 g, 37.6 mmol, 1.4 equiv) and triphosgene (3.19 g, 10.7 mmol, 0.4 equiv). The reaction mixture was warmed to room temperature and stirred for 2 h. The progress of the reaction was monitored by TLC. After completion of the starting material, the reaction mixture was washed with water (100 mL) and extracted with DCM (2 × 100 mL). The combined organic extracts were washed again with water (200 mL), brine (200 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate as a crude oil (6.0 g, 90%).

[0671] 1 1H NMR (400 MHz, DMSO-d6) δ 3.64 (br d, J = 4.40 Hz, 2H), 3.52 (br s, 2H), 3.27 - 3.47 (m, 3H), 2.93 - 3.21 (m, 1H), 1.41 (s, 9H).

[0672] Step 1-2: Preparation of (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate · 4-(10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 1)

[0673] At 0 °C, DIPEA (15.3 g, 118 mmol, 5 equiv) and DMAP (724 mg, 5.9 mmol, 0.25 equiv) were added to a stirred solution of (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-1, 10 g, 23.7 mmol, 1.0 equiv) in DCM (100 mL, 10 vol). Subsequently, tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate (5.89 g, 23.7 mmol, 1 equiv) in DCM (100 mL, 10 vol) was added dropwise over 10 min. The resulting reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the mixture was washed with water (500 mL), extracted with DCM (3 × 100 mL), and the combined organic extracts were washed again with water (200 mL) and brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The obtained crude product was purified by combiflash column eluting with 7% methanol / DCM to give Intermediate 1 (10 g, 66%) as an off-white solid.

[0674] 1 H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.11 (d, J = 9.29 Hz, 1H), 7.65 (d, J = 9.29 Hz, 1H), 7.34 (s, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 3.67 - 3.79 (m, 4H), 3.41 - 3.55 (m, 6H), 2.20 (s, 6H), 1.80 - 1.93 (m, 2H), 1.44 (s, 9H), 0.89 (br t, J = 7.34 Hz, 3H). LCMS: 634.2 [M+H] + 。

[0675] Step 2: Preparation of (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate · 4-(10-((dimethylamino)methyl)-4-((dimethylglycyl)oxy)-4-ethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2)

[0676] At 0 °C under an argon atmosphere, dimethylglycine (SM-3, 1.7 g, 16.57 mmol, 1.5 equiv), DCC (3.41 g, 16.57 mmol, 1.5 equiv) and DMAP (134 mg, 1.105 mmol, 0.1 equiv) were added to a stirred solution of piperazine-1,4-dicarboxylic acid (S)-1-(tert-butyl) ester·4-(10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 1, 7.0 g, 11.05 mmol, 1.0 equiv) in DCM (70 mL, 10.0 vol). The reaction mixture was stirred at ambient temperature until TLC indicated complete consumption of the starting material. The reaction mixture was then diluted with ice water (250 mL) and extracted with DCM (2 × 250 mL). The combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product obtained was purified by flash column chromatography, the pure fractions were combined and concentrated under reduced pressure to give piperazine-1,4-dicarboxylic acid (S)-1-(tert-butyl) ester·4-(10-((dimethylamino)methyl)-4-((dimethylglycyl)oxy)-4-ethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2, 6 g, 75%) as an off-white solid.

[0677] 1 H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.09 (d, J = 8.31 Hz, 1H), 7.65 (d, J = 9.29 Hz, 1H), 7.06 (s, 1H), 5.41 - 5.59 (m, 2H), 5.26 - 5.39 (m, 2H), 3.76 (s, 2H), 3.65 - 3.73 (m, 2H), 3.40 - 3.56 (m, 5H), 3.23 - 3.31 (m, 1H), 3.07 - 3.15 (m, 1H), 2.94 (s, 2H), 2.24 - 2.28 (m, 4H), 2.19 - 2.22 (m, 4H), 2.11 - 2.18 (m, 1H), 1.55 - 1.76 (m, 1H), 1.44 (s, 9H), 1.40 (s, 3H), 0.93 (t, J = 7.09 Hz, 3H). LCMS: 719.5 [M+H] + 。

[0678] Step 3: Preparation of (S)-10-((dimethylamino)methyl)-4-((dimethylglycyl)oxy)-4-ethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate 3)

[0679] To a stirred solution of (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate · 4-(10-((dimethylamino)methyl)-4-((dimethylglycyl)oxy)-4-ethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2, 5 g, 6.96 mmol, 1.0 equiv) in DCM (50 mL, 10 vol) at ambient temperature was added TFA (5.32 mL, 69.6 mmol, 10.0 equiv). The reaction mixture was stirred at room temperature until TLC indicated complete consumption of the starting material. The reaction mixture was then concentrated under reduced pressure, diluted with DCM (500 mL), washed with saturated bicarbonate solution (2 × 250 mL), brine (250 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford (S)-10-((dimethylamino)methyl)-4-((dimethylglycyl)oxy)-4-ethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate 3, 2.2 g, 51%) as a pale yellow solid.

[0680] 1 H NMR (400 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.08 (d, J = 9.26 Hz, 1H), 7.63 (d, J = 9.26 Hz, 1H), 7.06 (s, 1H), 5.50 (s, 2H), 5.32 (s, 2H), 3.76 (s, 2H), 3.63 (s, 2H), 3.37 - 3.47 (m, 4H), 2.72 - 2.87 (m, 5H), 2.55 - 2.64 (m, 1H), 2.26 (s, 5H), 2.20 (s, 6H), 2.16 (dd, J = 7.44, 2.19 Hz, 2H), 0.93 (t, J = 7.38 Hz, 3H). LCMS: 619.3 [M+H] + .

[0681] Step 4: Preparation of (S)-10-((dimethylamino)methyl)-4-((dimethylglycyl)oxy)-4-ethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylate

[0682] At 0 °C under an argon atmosphere, acetic acid (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(6-oxohexyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl ester (Intermediate C, 2.26 g, 4.04 mmol) and acetic acid (1 mL, catalytic amount) were added to a stirred solution of (S)-10-((dimethylamino)methyl)-4-((dimethylglycyl)oxy)-4-ethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate (Intermediate 3, 2.5 g, 4.04 mmol, 1.0 equiv) in MeOH (25 mL, 10 vol). The reaction mixture was stirred at room temperature for 1 h, and then NaCNBH3 (509 mg, 8.08 mmol) was added at 0 °C under an argon atmosphere. The resulting reaction mixture was stirred at room temperature until TLC showed complete consumption of the starting materials. The reaction mixture was quenched with saturated sodium bicarbonate solution (250 mL) and extracted with ethyl acetate (2 × 500 mL). The combined organic layers were washed with brine solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was purified by preparative HPLC (column: spherical-C18, 40 um, 100 A; mobile phase A: 0.1% FA / water; mobile phase B: acetonitrile; flow rate: 25 mL / min, diluent: THF + DMSO). The pure fractions were combined and lyophilized under reduced pressure to obtain 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-((dimethylglycyl)oxy)-4-ethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (663 mg, 14%).

[0683] 11H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.09 (d, J = 9.29 Hz, 1H), 7.63 (d, J = 9.29 Hz, 1H), 7.06 (s, 1H), 6.99 (d, J = 8.31 Hz, 2H), 6.59 (d, J = 8.31 Hz, 2H), 5.67 (s, 1H), 5.50 (s, 2H), 5.30 - 5.35 (m, 2H), 4.38 - 4.43 (m, 1H), 3.67 - 3.77 (m, 4H), 3.37 - 3.47 (m, 6H), 3.22 - 3.27 (m, 4H), 2.82 (s, 3H), 2.61 - 2.82 (m, 3H), 2.53 - 2.58 (m, 2H), 2.41 (s, 2H), 2.30 - 2.37 (m, 3H), 2.26 (s, 5H), 2.16 - 2.21 (m, 10H), 2.10 (s, 4H), 2.00 (s, 4H), 1.86 - 1.98 (m, 2H), 1.62 - 1.77 (m, 1H), 1.39 - 1.52 (m, 5H), 1.26 - 1.36 (m, 5H), 0.90 - 0.95 (m, 3H), 0.24 (s, 3H). LCMS: 1162.1 [M + H] + 。

[0684] Example S15: Preparation of N-((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)-6-(4-(4-(((S)-4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)-piperazine-1-carbonyl)piperidin-1-yl)pyridazine-3-carboxamide (Compound No. 15)

[0685]

[0686] (S)-4-Ethyl-4,9-dihydroxy-10-(piperazin-1-ylmethyl)-1,12-dihydro-14H-pyrano[3',4':6,7]-indazino[1,2-b]quinoline-3,14(4H)-dione trifluoroacetate (Intermediate B) Preparation

[0687] Step B1: Preparation of (S)-4-((4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (Intermediate B1)

[0688] Under an inert atmosphere at room temperature, 37% formaldehyde solution (0.29 mL, 8.24 mmol, 1.2 equiv) was added to a stirred solution of (S)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-1, 2.5 g, 6.86 mmol, 1.0 equiv) and tert-butyl piperazine-1-carboxylate (SM-2, 1.85 g, 10.3 mmol, 1.5 equiv) in acetic acid (10 mL). The reaction mixture was heated to 80 °C in a sealed tube for 2 h. The reaction progress was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure to obtain the crude product, which was basified to pH 9 with ammonia water. The resulting solid was filtered, washed with water (10 mL), and dried in vacuo to give Intermediate B1 (1.9 g, 50%) as a yellow solid.

[0689] LCMS: 463.46 [M - 100] + (Boc group cleavage was observed in LCMS).

[0690] Step B2: Preparation of (S)-4-ethyl-4,9-dihydroxy-10-(piperazin-1-ylmethyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione trifluoroacetate (Intermediate B)

[0691] Under a nitrogen atmosphere at 0 °C, TFA (2.7 mL, 35 mmol, 10 equiv) was added to a stirred solution of (S)-4-((4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-10-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (Intermediate B1, 2 g, 3.5 mmol, 1.0 equiv) in DCM (20 mL). The reaction mixture was warmed to room temperature and stirred for 2 h. The reaction progress was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, and the resulting residue was washed with ether (10 mL) and dried in vacuo to give Intermediate B (1.4 g, 83%) as a yellow solid.

[0692] 11H NMR (400 MHz, DMSO-d6) δ 8.86 (br s, 2H), 8.10 (d, J = 9.25 Hz, 1H), 7.58 (d, J = 9.25 Hz, 1H), 7.21 - 7.32 (m, 2H), 6.97 - 7.15 (m, 1H), 6.34 - 6.64 (m, 1H), 5.42 (s, 2H), 5.26 (s, 2H), 4.40 (s, 2H), 3.10 - 3.30 (m, 8H), 1.85 - 1.89 (m, 2H), 0.88 (t, J = 7.17 Hz, 3H).

[0693] Preparation of N-((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)-6-(4-(4-(((S)-4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)-piperazine-1-carbonyl)piperidin-1-yl)pyridazine-3-carboxamide

[0694] At room temperature, HOBt (138 mg, 0.90 mmol, 1.5 equiv), EDCI·HCl (173 mg, 0.90 mmol, 1.5 equiv) and DIPEA (155.7 mg, 1.20 mmol, 2 equiv) were added to a stirred solution of 1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamoyl)-pyridazin-3-yl)piperidine-4-carboxylic acid (Intermediate 6 of Example S11, 300 mg, 0.60 mmol, 1.0 equiv) and (S)-4-ethyl-4,9-dihydroxy-10-(piperazin-1-ylmethyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione trifluoroacetate (Intermediate B, 278 mg, 0.60 mmol, 1.0 equiv) in DCM (10 mL), and the mixture was stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (50 mL) was added and the aqueous solution was extracted with DCM (2 × 100 mL). The combined organic extracts were washed with water (100 mL) and brine (100 mL) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (in a mobile phase of ammonium bicarbonate / water and acetonitrile) to give the title compound as an off-white solid (60 mg, 10%).

[0695] 11H NMR (400 MHz, DMSO-d6) δ 8.77 (s, 1H), 8.48 (d, J = 8.26 Hz, 1H), 7.99 (d, J = 9.26 Hz, 1H), 7.82 (d, J = 9.51 Hz, 1H), 7.60 (d, J = 9.01 Hz, 1H), 7.47 (d, J = 9.13 Hz, 1H), 7.35 (d, J = 9.76 Hz, 1H), 7.26 (s, 1H), 6.94 (d, J = 2.38 Hz, 1H), 6.83 (dd, J = 9.07, 2.44 Hz, 1H), 6.48 (s, 1H), 5.42 (s, 2H), 5.26 (s, 2H), 4.48 (br d, J = 13.01 Hz, 2H), 4.03 (s, 2H), 3.75 - 3.88 (m, 2H), 3.41 - 3.62 (m, 3H), 2.96 - 3.16 (m, 3H), 2.85 (s, 3H), 2.51 - 2.54 (m, 2H), 1.83 - 1.97 (m, 5H), 1.51 - 1.79 (m, 12H), 0.88 (t, J = 7.32 Hz, 3H). (No 1H exchangeable hydrogen was observed in the spectrum). LCMS: 941.4 [M+H] + . HPLC: 94.2%.

[0696] Example S16: Preparation of N-((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)-6-(4-(4-((((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl)oxy)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyridazine-3-carboxamide (Compound No. 16)

[0697]

[0698] Step 1: Preparation of tert-butyl (S)-4-(((10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl)oxy)methyl)piperidine-1-carboxylate (Intermediate 1)

[0699] Under a nitrogen atmosphere at room temperature, potassium carbonate (1.31 g, 9.5 mmol, 2.0 equiv) and tert-butyl 4-(bromomethyl)piperidine-1-carboxylate (SM-2, 1.71 g, 6.17 mmol, 1.3 equiv) were added to a stirred solution of (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-1, 2 g, 4.75 mmol, 1.0 equiv) in DMF (10 mL). The reaction mixture was heated to 50 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with ice water (20 mL) and extracted with ethyl acetate (2 × 50 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give Intermediate 1 (1.0 g, crude) as a yellow solid, which was used in the next step without further purification.

[0700] LCMS: 617.2 [M-H] - 。

[0701] Step 2: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-9-(piperidin-4-ylmethoxy)-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione trifluoroacetate (Intermediate 2)

[0702] At 0 °C, TFA (10 mL) was added to a stirred solution of Intermediate 1 (1.0 g, 1.41 mmol, 1.0 equiv) in DCM (50 mL). The reaction mixture was warmed to room temperature and stirred for 12 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure to give the crude product, which was triturated with ether:pentane (80:20) three times (25 mL each time) and dried in vacuo to give Intermediate 2 (800 mg, crude) as a pale yellow solid, which was used in the next step without further purification.

[0703] LCMS: 519.26 [M+H] + 。

[0704] Preparation of N-((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)-6-(4-(4-((((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl)oxy)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyridazine-3-carboxamide

[0705] To a stirred solution of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-9-(piperidin-4-ylmethoxy)-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione (Intermediate 2, 370 mg, 0.772 mmol, 1.0 equiv) in DMF (3 mL) was added DIPEA (0.27 mL, 1.54 mmol, 2.0 equiv) and HATU (440 mg, 1.15 mmol, 1.5 equiv). After 5 - 10 minutes, Intermediate 13 (400 mg, 0.772 mmol, 1.0 equiv) was added at room temperature and stirring was continued for an additional 16 h. The reaction progress was monitored by TLC. After completion of the reaction, the solvent was evaporated and the resulting residue was triturated with ether (20 mL), filtered, dried and purified by preparative HPLC elution (mobile phase A: 0.1% TFA / water, mobile phase B: acetonitrile) to give the title compound as a white solid (50 mg, 6%).

[0706] 1 1H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1H), 8.58 (d, J = 8.13 Hz, 1H), 8.31 (s, 2H), 8.07 - 8.16 (m, 1H), 7.69 - 7.90 (m, 2H), 7.29 - 7.41 (m, 2H), 7.07 - 7.22 (m, 1H), 6.40 - 6.54 (m, 1H), 5.37 - 5.45 (m, 2H), 5.21 - 5.31 (m, 2H), 4.43 - 4.58 (m, 4H), 4.05 - 4.23 (m, 3H), 3.86 (brs, 3H), 3.06 - 3.20 (m, 5H), 2.62 (br s, 1H), 2.21 (s, 6H), 2.07 - 2.14 (m, 2H), 1.79 - 1.99 (m, 6H), 1.70 - 1.79 (m, 2H), 1.47 - 1.70 (m, 6H), 1.39 (d, J = 10.88 Hz, 1H), 1.21 - 1.30 (m, 1H), 0.82 - 0.95 (m, 3H). LCMS: 984.3 [M+H] + . HPLC purity 96.3%.

[0707] Example S17: Preparation of (2R,5S)-4-(1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (Compound 17)

[0708]

[0709] Step 1: Preparation of (2S,5R)-4-(chlorocarbonyl)-2,5-dimethylpiperazine-1-carboxylic acid tert-butyl ester (Intermediate 1)

[0710] To a stirred solution of (2S,5R)-2,5-dimethylpiperazine-1-carboxylic acid tert-butyl ester (SM-1, 3.0 g, 13.99 mmol, 1.0 equiv) in DCM (25 mL) at 0 °C was added dropwise a solution of pyridine (1.7 mL, 20.99 mmol, 1.5 equiv) and triphosgene (1.24 g, 41.99 mmol, 0.3 equiv) in DCM (5 mL) over 10 min. The reaction mixture was warmed to room temperature and stirred for 30 min. The progress of the reaction was monitored by TLC (non-polar spots were observed). After completion of the reaction, the reaction mixture was poured into ice water (50 mL) and extracted with DCM (2 x 30 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give Intermediate 1 (3.7 g, crude) as a pale brown gum, which was used in the next step without further purification.

[0711] 1 H NMR (400 MHz, DMSO-d6) δ 4.09 - 4.55 (m, 2H), 3.46 - 3.69 (m, 2H), 3.13 - 3.41 (m, 2H), 1.40 (s, 9H), 0.98 - 1.28 (m, 6H).

[0712] Step 2: Preparation of (2S,5R)-2,5-dimethylpiperazine-1,4-dicarboxylic acid 1-(tert-butyl) ester · 4-((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2)

[0713] At 0 °C, DIPEA (5.67 mL, 32.6 mmol, 5 equiv) was added to a solution of (2S,5R)-tert-butyl 4-(chlorocarbonyl)-2,5-dimethylpiperazine-1-carboxylate (Intermediate 1, 3.6 g, 13.05 mmol, 2 equiv) and (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-2, 3.1 g, 6.52 mmol, 1 equiv) in THF (30 mL) and DMF (30 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with water (30 mL) and extracted with ethyl acetate (2 × 25 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash column chromatography eluting with 10% methanol / DCM to give Intermediate 2 as a pale yellow foam (2.5 g, 58%).

[0714] 1 H NMR (400 MHz, DMSO-d6) δ 8.95 (br s, 1H), 8.11 (d, J = 9.29 Hz, 1H), 7.54 - 7.76 (m, 1H), 7.34 (s, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 4.19–4.41 (m, 2H), 3.61 - 3.81 (m, 2H), 3.17 (d, J = 12.91 Hz, 1H), 2.20 (d, J = 6.36 Hz, 6H), 1.80 - 1.95 (m, 3H), 1.26 - 1.41 (m, 14H), 1.18 (d, J = 6.36 Hz, 3H), 0.89 (t, J = 6.36 Hz, 3H). LCMS: 662.4 [M+H] + 。

[0715] Step 3: Preparation of (2R,5S)-tert-butyl 2,5-dimethylpiperazine-1-carboxylate (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (Intermediate 3)

[0716] At 0 °C, TFA (3 mL) was added to a stirred solution of (2S,5R)-2,5-dimethylpiperazine-1,4-dicarboxylic acid 1-(tert-butyl) ester·4-((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate 2, 1.5 g, 2.26 mmol, 1.0 equiv) in DCM (20 mL). The reaction mixture was warmed to room temperature and stirred for 5 h / 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, saturated NaHCO3 (60 mL) was added, and the aqueous solution was extracted with ethyl acetate (2 × 100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give Intermediate 3 as a pale yellow solid (1.20 g, 78%).

[0717] 1 H NMR (400 MHz, DMSO-d6) δ 10.01 (br s, 1H), 9.08 (br s, 3H), 8.32 (d, J = 9.00 Hz, 1H), 7.80 (d, J = 9.39 Hz, 1H), 7.34 (s, 1H), 6.53 (br s, 1H), 5.41 (s, 2H), 5.30 (s, 2H), 4.79 (br s, 2H), 3.91 (br s, 2H), 3.69 (br s, 2H), 3.11 (d, J = 12.91 Hz, 1H), 2.84 (br s, 6H), 1.86 - 1.92 (m, 2H), 1.38 (br s, 6H), 1.21 (d, J = 6.65 Hz, 1H), 0.85 (t, J = 7.24 Hz, 3H). LCMS: 562.5 [M+H] + 。

[0718] Step 4: Preparation of (2R,5S)-4-(1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester

[0719] At room temperature, HATU (323 mg, 0.85 mmol, 1.5 eq) and DIPEA (0.20 mL, 1.5 mmol, 2 eq) were added to a stirred solution of (2R,5S)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (Intermediate 3, 380 mg, 0.67 mmol, 1.2 eq) and 1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)-carbamoyl)pyridazin-3-yl)piperidine-4-carboxylic acid (Intermediate 6, 280 mg, 0.56 mmol, 1.0 eq) in DMF (3 mL), and the mixture was allowed to stir for an additional 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (50 mL) was added, and the aqueous solution was extracted with ethyl acetate (2 × 200 mL). The combined organic extracts were washed with water (100 mL), brine (100 mL) and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (in ammonium bicarbonate / water and acetonitrile mobile phases) to give the title compound as an off-white solid (108 mg, 37%).

[0720] 1 H NMR (400 MHz, DMSO-d6) δ 8.96 (s, 1H), 8.49 (br d, J = 8.25 Hz, 1H), 8.13 (br d, J = 9.01 Hz, 1H), 7.84 (d, J = 9.63 Hz, 1H), 7.55 - 7.73 (m, 2H), 7.29 - 7.46 (m, 2H), 6.95 (d, J = 2.38 Hz, 1H), 6.83 (dd, J = 9.07, 2.31 Hz, 1H), 6.52 (s, 1H), 5.43 (s, 2H), 5.32 (s, 2H), 4.70 - 4.79 (m, 1H), 4.42 - 4.64 (m, 3H), 4.16 - 4.38 (m, 1H), 3.57 - 4.03 (m, 6H), 3.10 - 3.14 (m, 3H), 2.85 (s, 3H), 2.21 (d, J = 6.82 Hz, 6H), 1.82 - 1.99 (m, 5H), 1.60 - 1.81 (m, 9H), 1.42 (br dd, J = 10.51, 6.75 Hz, 2H), 1.32 (br d, J = 6.75 Hz, 1H), 1.21 - 1.29 (m, 3H), 1.12 (br dd, J = 10.38, 6.88 Hz, 1H), 0.89 (t, J = 7.32 Hz, 3H). LCMS: 1041.7 [M+H]+ 。HPLC: 99.42%.

[0721] Example S18: Preparation of (S)-10-((Dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-((1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)methyl)piperazine-1-carboxylate (Compound No. 18)

[0722]

[0723] At room temperature, piperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl ester (Intermediate A, 312 mg, 0.58 mmol, 1.0 equiv) and acetic acid (0.1 mL) were added to a stirred solution of N-((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)-6-(4-formylpiperidin-1-yl)pyridazine-3-carboxamide (Intermediate 6, 280 mg, 0.58 mmol, 1.0 equiv) in methanol (5 mL), and the mixture was stirred for 2 h. At 0 °C, NaCNBH3 (58 mg, 0.9 mmol, 1.5 equiv) was added portionwise to the reaction mixture. The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, cold water (50 mL) was added and the mixture was extracted with ethyl acetate (2 × 200 mL). The combined organic extracts were washed with water (100 mL), brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash column eluting with 8% methanol / DCM to give the title compound as an off-white solid (125 mg, 23%).

[0724] 11H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.51 (br d, J = 8.31 Hz, 1H), 8.12 (d, J = 8.80 Hz, 1H), 7.82 (d, J = 9.29 Hz, 1H), 7.59 - 7.66 (m, 2H), 7.31 - 7.38 (m, 2H), 6.95 (br s, 1H), 6.83 (br d, J = 9.29 Hz, 1H), 6.54 (s, 1H), 5.44 (s, 2H), 5.32 (br s, 2H), 4.48 - 4.52 (m, 2H), 3.70 - 3.81 (m, 5H), 3.47 - 3.50 (m, 2H), 2.99 - 3.08 (m, 3H), 2.85 (s, 3H), 2.48 - 2.51 (m, 4H) 2.20 - 2.22 (m, 2H), 2.19 (s, 6H) 1.82 - 1.96 (m, 6H), 1.62 - 1.79 (m, 5H), 1.11 - 1.23 (m, 4H), 0.88 (br t, J = 6.85 Hz, 3H). LCMS: 998.2 [M+H] + . HPLC purity: 96.8%.

[0725] Example S19: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((2-(4-((((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl)oxy)methyl)-piperidin-1-yl)-2-oxoethyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Compound No. 19)

[0726]

[0727] Step 1: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methylamino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1)

[0728] At 0 °C, potassium acetate (20.6 g, 210 mmol, 10 equiv) and iodine (13.1 g, 105 mmol, 5 equiv) were added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-(dimethylamino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (SM-1, 10 g, 21 mmol, 1.0 equiv) in methanol (150 mL) and THF (150 mL). The reaction mixture was warmed to room temperature and stirred for 3 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with sodium thiosulfate (Na2S2O3) (50 g in 30 mL of water) and extracted with ethyl acetate (2 × 200 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give Intermediate 1 as an off-white solid (8.0 g, 82%), which was used in the next step without further purification.

[0729] 1 H NMR (400 MHz, DMSO-d6) δ 11.91 (br s, 1H), 6.91 (d, J = 8.31 Hz, 2H), 6.44 (d, J = 8.31 Hz, 2H), 5.67 (s, 1H), 4.37 (m, 1H), 2.75 (s, 2H), 2.61 (d, J = 4.40 Hz, 3H), 2.30 - 2.40 (m, 1H), 2.07 - 2.16 (s, 5H), 1.99 (s, 6H), 1.63 - 1.77 (m, 2H), 1.21 - 1.45 (m, 5H), 0.86 (t, J = 6.60 Hz, 1H), 0.16 - 0.28 (m, 3H). LCMS: 462.28 [M+H] + 。

[0730] Step 2: Preparation of tert-butyl N-(4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylglycinate (Intermediate 2)

[0731] At room temperature, SM-2 (0.32 mL, 2.17 mmol, 1 equiv) and NaHCO3 (911 mg, 10.85 mmol, 5 equiv) were added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methylamino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate 1, 1 g, 2.17 mmol, 1 equiv) in EtOH:H2O (20 mL, 1:1). The reaction mixture was heated to 80 °C and stirred for 2 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with cold water (50 mL) and extracted with ethyl acetate (2 × 50 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product. The obtained crude product was triturated with n-heptane (2 × 30 mL), filtered, and dried in vacuo to give Intermediate 2 as a pale yellow solid (900 mg, 72%).

[0732] 1 H NMR (400 MHz, DMSO-d6) δ 7.00 (d, J = 8.31 Hz, 2H), 6.56 (d, J = 8.31 Hz, 2H), 5.67 (s, 1H), 5.67 (s, 1H), 4.41 (d, J = 6.85 Hz, 1H), 4.00 (s, 2H), 2.91 (s, 3H), 2.58 - 2.82 (m, 3H), 2.54 - 2.57 (m, 1H), 2.29 - 2.38 (m, 1H), 2.06 - 2.23 (m, 6H), 1.84 - 2.03 (m, 5H), 1.61 - 1.79 (m, 2H), 1.34 - 1.46 (m, 3H), 1.30 (s, 9H), 0.22 (s, 3H). LCMS: 576.2 [M+H] + 。

[0733] Step 3: Preparation of N-(4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylglycine (Intermediate 3)

[0734] At 0 °C, trimethylchlorosilane (1.31 mL, 10.43 mmol, 20 equiv) was added dropwise to a stirred solution of tert-butyl N-(4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylglycinate (Intermediate 2, 300 mg, 0.521 mmol, 1 equiv) in trifluoroethanol (5 mL). The reaction mixture was warmed to room temperature and stirred for 4 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated, and the resulting residue was washed with pentane (2 × 20 mL) and dried in vacuo to give Intermediate 3 as a pale yellow solid (250 mg, 92%).

[0735] 1 H NMR (400 MHz, DMSO-d6) δ 6.99 (d, J = 8.50 Hz, 2H), 6.55 (d, J = 8.76 Hz, 2H), 5.67 (s, 1H), 4.40 (d, J = 6.63 Hz, 1H), 4.01 (s, 2H), 2.92 (s, 2H), 2.59 - 2.79 (m, 3H), 2.55 (br s, 2H), 2.32 - 2.39 (m, 1H), 2.02–2.22 (m, 1H), 2.13 - 2.18 (m, 2H), 2.09 (s, 3H), 1.99 (s, 4H), 1.87 - 1.97 (m, 2H), 1.65–1.75 (m, 2H), 1.33 - 1.40 (m, 2H), 1.24 - 1.32 (m, 3H), 0.84 - 0.87 (m, 2H). LCMS: 520.2 [M+H] + 。

[0736] Step 4: Preparation of (8S,11R,13S,14S,17R)-17-acetyl-11-(4-((2-(4-((((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl)oxy)methyl)piperidin-1-yl)-2-oxoethyl)(methyl)amino)phenyl)-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate

[0737] At room temperature, to a stirred solution of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-9-(piperidin-4-ylmethoxy)-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione TFA salt (Intermediate C, 350 mg, 0.675 mmol, 1.0 equiv) and N-(4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)-N-methylglycine (Intermediate 3, 350 mg, 0.675 mmol, 1.0 equiv) in DMF (4 mL) was added DIPEA (0.35 mL, 2.02 mmol, 3.0 equiv) and HATU (384 mg, 1.01 mmol, 1.5 equiv), and stirring was continued for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted in cold water (30 mL), the resulting solid was filtered, and purification was carried out by preparative HPLC eluting with mobile phase A: 0.1% FA / water and mobile phase B: acetonitrile to give the title compound as an off-white solid (24 mg, 3%).

[0738] 1 H NMR (400 MHz, DMSO-d6) δ 9.34 (br s, 1H), 8.94 (s, 1H), 8.33 (d, J = 9.29 Hz, 1H), 7.91 (d, J = 9.29 Hz, 1H), 7.30 (s, 1H), 6.93 (d, J = 7.83 Hz, 2H), 6.51 (d, J = 8.80 Hz, 3H), 5.64 (s, 1H), 5.41 (s, 2H), 5.25 (s, 2H), 4.75 (br s, 2H), 4.36 (br s, 2H), 4.10 - 4.24 (m, 4H), 3.86 - 3.97 (m, 2H), 2.88 (br s, 9H), 2.51–2.55 (m, 3H), 2.18–2.59 (m, 2H), 2.07 (s, 8H), 1.96 (s, 9H), 1.60 - 1.73 (m, 3H), 1.24 - 1.41 (m, 5H), 0.86 (t, J = 6.85 Hz, 3H), 0.20 (s, 2H). LCMS: 1020.5 [M+H] + . HPLC purity 94.3%.

[0739] Example S20: Preparation of (2R,5S)-4-((1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)-(methyl)amino)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)methyl)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (Compound No. 20)

[0740]

[0741] Step D1: Preparation of tert-butyl (2S,5R)-4-(chlorocarbonyl)-2,5-dimethylpiperazine-1-carboxylate (Intermediate D1)

[0742] To a stirred solution of tert-butyl (2S,5R)-2,5-dimethylpiperazine-1-carboxylate (SM-1, 3.0 g, 13.99 mmol, 1.0 equiv) in DCM (25 mL) at 0 °C was added dropwise a solution of pyridine (1.7 mL, 20.99 mmol, 1.5 equiv) and triphosgene (1.24 g, 41.99 mmol, 0.3 equiv) in DCM (5 mL) over 10 min. The reaction mixture was warmed to room temperature and stirred for 30 min. The progress of the reaction was monitored by TLC (non-polar spots were observed). After completion of the reaction, the reaction mixture was poured into ice water (50 mL) and extracted with DCM (2 x 30 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give Intermediate D1 (3.7 g, crude) as a light brown gum, which was used in the next step without further purification.

[0743] 1 1H NMR (400 MHz, DMSO-d6) δ 4.09 - 4.55 (m, 2H), 3.46 - 3.69 (m, 2H), 3.13 - 3.41 (m, 2H), 1.40 (s, 9H), 0.98 - 1.28 (m, 6H).

[0744] Step D2: Preparation of 1-(tert-butyl) (2S,5R)-2,5-dimethylpiperazine-1,4-dicarboxylate · 4-((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate D2)

[0745] At 0 °C, DIPEA (5.67 mL, 32.6 mmol, 5 eq) was added to a solution of (2S,5R)-tert-butyl 4-(chlorocarbonyl)-2,5-dimethylpiperazine-1-carboxylate (Intermediate D1, 3.6 g, 13.05 mmol, 2 eq) and (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-2, 3.1 g, 6.52 mmol, 1 eq) in THF (30 mL) and DMF (30 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with water (30 mL) and extracted with ethyl acetate (2 × 25 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash column chromatography eluting with 10% methanol / DCM to give Intermediate D2 as a pale yellow foam (2.5 g, 58%).

[0746] 1 H NMR (400 MHz, DMSO-d6) δ 8.95 (br s, 1H), 8.11 (d, J = 9.29 Hz, 1H), 7.54 - 7.76 (m, 1H), 7.34 (s, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 4.19–4.41 (m, 2H), 3.61 - 3.81 (m, 2H), 3.17 (d, J = 12.91 Hz, 1H), 2.20 (d, J = 6.36 Hz, 6H), 1.80 - 1.95 (m, 3H), 1.26 - 1.41 (m, 14H), 1.18 (d, J = 6.36 Hz, 3H), 0.89 (t, J = 6.36 Hz, 3H). LCMS: 662.4 [M+H] + 。

[0747] Step D3: Preparation of (2R,5S)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (Intermediate D)

[0748] At 0 °C, trifluoroacetic acid (TFA, 3 mL) was added to a stirred solution of (2S,5R)-2,5-dimethylpiperazine-1,4-dicarboxylic acid 1-(tert-butyl) ester 4-((S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate D2, 1.5 g, 2.26 mmol, 1.0 equiv) in DCM (20 mL). The reaction mixture was warmed to room temperature and stirred for 5 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, saturated NaHCO3 solution (50 mL) was added, and the aqueous solution was extracted with EtOAc (2 × 60 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give Intermediate D as a pale yellow solid (1.20 g, 78%).

[0749] 1 1H NMR (400 MHz, DMSO-d6) δ 10.01 (br s, 1H), 9.08 (br s, 3H), 8.32 (d, J = 9.00 Hz, 1H), 7.80 (d, J = 9.39 Hz, 1H), 7.34 (s, 1H), 6.53 (br s, 1H), 5.41 (s, 2H), 5.30 (s, 2H), 4.79 (br s, 2H), 3.91 (br s, 2H), 3.69 (br s, 2H), 3.11 (d, J = 12.91 Hz, 1H), 2.84 (br s, 6H), 1.86 - 1.92 (m, 2H), 1.38 (br s, 6H), 1.21 (d, J = 6.65 Hz, 1H), 0.85 (t, J = 7.24 Hz, 3H). LCMS: 562.5 [M+H] + 。

[0750] Step 1: Preparation of N-((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)-6-(4-(hydroxymethyl)piperidin-1-yl)pyridazine-3-carboxamide (Intermediate 5)

[0751] At room temperature, piperidin-4-ylmethanol (SM-3, 0.690 g, 6 mmol, 1.2 equiv) and K2CO3 (1.1 g, 8 mmol, 1.6 equiv) were added to a stirred solution of intermediate 4 (2.0 g, 5 mmol, 1.0 equiv) in DMF (10 mL). The reaction mixture was heated to 90 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, cold water (50 mL) was added to the reaction mixture and the mixture was extracted with ethyl acetate (2 × 150 mL). The combined organic layer extracts were washed with water (100 mL), brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash column eluting with 100% ethyl acetate to give intermediate 5 (1.3 g, 54%) as an off-white solid.

[0752] 1 H NMR (400 MHz, DMSO-d6) δ 8.49 (br d, J = 8.31 Hz, 1H), 7.80 (d, J = 9.78 Hz, 1H), 7.61 (d, J = 8.80 Hz, 1H), 7.33 (d, J = 9.78 Hz, 1H), 6.94 (d, J = 1.96 Hz, 1H), 6.78 - 6.86 (m, 1H), 4.42 - 4.55 (m, 3H), 3.72 - 3.92 (m, 2H), 3.20 - 3.27 (m, 3H), 2.98 (br t, J = 12.47 Hz, 2H), 2.85 (s, 3H), 1.91 (br d, J = 9.29 Hz, 2H), 1.64 - 1.78 (m, 8H), 1.08 - 1.21 (m, 2H). LCMS: 483.2 [M+H] + 。

[0753] Step 2: Preparation of N-((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)-6-(4-formylpiperidin-1-yl)pyridazine-3-carboxamide (Intermediate 6)

[0754] Under a nitrogen atmosphere at 0 °C, Dess-Martin periodinane (400 mg, 0.92 mmol, 1.3 equiv) was added portionwise to a stirred solution of N-((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)-6-(4-(hydroxymethyl)piperidin-1-yl)pyridazine-3-carboxamide (Intermediate 5, 350 mg, 0.72 mmol, 1.0 equiv) in DCM (6 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, water (50 mL) was added and the aqueous reaction mixture was extracted with DCM (2 × 50 mL). The combined organic extracts were washed with saturated NaHCO3 (50 mL) and saturated sodium thiosulfate (50 mL) solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product Intermediate 6 (320 mg, 91%) as an off-white solid, which was used in the next step without purification.

[0755] 1 H NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 8.51 (br d, J = 8.31 Hz, 1H), 7.82 (d, J = 9.78 Hz, 1H), 7.61 (d, J = 8.80 Hz, 1H), 7.29 - 7.41 (m, 1H), 6.94 (s, 1H), 6.82 (br d, J = 7.83 Hz, 1H), 4.31 (br d, J = 13.21 Hz, 1H), 3.98 - 4.08 (m, 1H), 3.72 - 3.91 (m, 1H), 3.21 - 3.31 (m, 2H), 2.85 (s, 3H), 2.65 - 2.68 (m, 1H), 1.86 - 2.03 (m, 4H), 1.60 - 1.82 (m, 5H), 1.45 - 1.60 (m, 2H), 1.17 (t, J = 7.09 Hz, 2H). LCMS: 481.2 [M+H] + 。

[0756] Step 3: Preparation of (2R,5S)-4-((1-(6-(((1r,4r)-4-((3-chloro-4-cyanophenyl)(methyl)amino)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)methyl)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester

[0757] At room temperature, to a stirred solution of (2R,5S)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (Intermediate 6, 320 mg, 0.66 mmol, 1.0 equiv) in methanol (5 mL) was added Intermediate D (486 mg, 0.86 mmol, 1.3 equiv) and acetic acid (0.1 mL), and the reaction mixture was stirred for an additional 2 h. At 0 °C, NaCNBH3 (64 mg, 1 mmol, 1.5 equiv) was added portionwise to the reaction mixture. The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, cold water (50 mL) was added and the mixture was extracted with ethyl acetate (2 × 200 mL). The combined organic extracts were washed with water (100 mL), brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The obtained crude product was purified by combiflash column eluting with 6% methanol / DCM to give the title compound as an off-white solid (369 mg, 54%).

[0758] 1 H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.48 (br d, J = 8.38 Hz, 1H), 8.11 (d, J = 9.13 Hz, 1H), 7.81 (d, J = 9.51 Hz, 1H), 7.57 - 7.65 (m, 2H), 7.31 - 7.38 (m, 2H), 6.95 (d, J = 2.38 Hz, 1H), 6.83 (dd, J = 9.13, 2.50 Hz, 1H), 6.51 (s, 1H), 5.28 - 5.46 (m, 4H), 4.43 - 4.57 (m, 2H), 3.71 - 3.89 (m, 5H), 2.96 - 3.11 (m, 4H), 2.85 (s, 3H), 2.80 - 2.82 (m, 2H), 2.41 (br d, J = 11.63 Hz, 1H), 2.20 - 2.35 (m, 3H), 2.14 (s, 6H), 1.58 - 2.03 (m, 14H), 1.30 - 1.45 (m, 2H), 1.10 - 1.22 (m, 1H), 0.96 - 1.12 (m, 3H), 0.89 (t, J = 7.32 Hz, 3H). LCMS: 1026.4 [M+H] + . HPLC purity: 94.5%.

[0759] Example S21: Preparation of (2R,5S)-4-(1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl ester formate (Compound No. 21)

[0760]

[0761] Step E1: Preparation of tert-butyl ((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamate (Intermediate E1)

[0762] Under a nitrogen atmosphere at 0 °C, NaH (4.08 g, 102 mmol, 2.2 equivalents) was added to a stirred solution of tert-butyl ((1r,4r)-4-hydroxycyclohexyl)carbamate (SM-2, 10 g, 46.5 mmol, 1.0 equivalent) in DMF (100 mL). The mixture was warmed to room temperature and stirred for 30 min. At room temperature, 2-chloro-4-fluoro-3-methylbenzonitrile (SM-1, 7.86 g, 46.51 mmol, 1.0 equivalent) was added portionwise to the reaction mixture over 10 min, and the resulting reaction mixture was stirred for 3 h. The progress of the reaction was monitored by TLC. After completion of the reaction, ice water (100 mL) was added, and the formed precipitate was filtered and dried to give Intermediate E1 as an off-white solid (15 g, 88%).

[0763] 1 1H NMR (400 MHz, DMSO-d6) δ 7.73 (d, J = 8.31 Hz, 1H), 7.22 (d, J = 8.80 Hz, 1H), 6.82 (s, 1H), 4.37 - 4.55 (m, 2H), 2.21 (s, 3H), 1.98 - 2.11 (m, 2H), 1.76 - 1.88 (m, 2H), 1.42 - 1.51 (m, 4H), 1.38 (s, 9H). LCMS: 309 [M - 56 + H] + 。

[0764] Step E2: Preparation of 4-(((1r,4r)-4-aminocyclohexyl)oxy)-2-chloro-3-methylbenzonitrile hydrochloride (Intermediate E2)

[0765] At 0 °C, 4 M HCl / 1,4-dioxane (75 mL) was added to a solution of intermediate E1 (15 g, 41.1 mmol) in DCM (25 mL). The reaction mixture was warmed to room temperature and stirred for 16 h. After completion of the reaction, the volatiles were evaporated under reduced pressure, and the resulting residue was washed with diethyl ether (2 × 80 mL) to give intermediate E2 (15 g, crude) as an off-white solid, which was used in the next step.

[0766] 1 H NMR (400 MHz, DMSO-d6) δ 8.30 (br s, 3H), 7.75 (d, J = 8.80 Hz, 1H), 7.28 (d, J = 8.80 Hz, 1H), 4.44 - 4.55 (m, 1H), 3.00 - 3.15 (m, 1H), 2.21 (s, 3H), 2.07 - 2.15 (m, 2H), 1.97 - 2.06 (m, 2H), 1.41 - 1.63 (m, 4H). LCMS: 264.75 [M+H] + 。

[0767] Step E3: Preparation of 6-chloro-N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)pyridazine-3-carboxamide (Intermediate E3)

[0768] At 0 °C, HATU (32.4 g, 85.44 mmol, 1.5 equiv) and DIPEA (49 mL, 284 mmol, 5.0 equiv) were added to a mixture of 4-(((1r,4r)-4-aminocyclohexyl)oxy)-2-chloro-3-methylbenzonitrile hydrochloride (Intermediate E2, 15 g, 56.9 mmol, 1.0 equiv) and 6-chloropyridazine-3-carboxylic acid (SM-3, 9 g, 56.9 mmol, 1.0 equiv) in DMF (100 mL). The reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the volatiles were evaporated under reduced pressure, diluted with water (500 mL), and extracted with ethyl acetate (3 × 300 mL). The combined organic extracts were washed with water (400 mL), brine (400 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a crude product, which was purified by column chromatography eluting with 60 - 80% ethyl acetate / hexane to give intermediate E3 (6 g, 27%) as an off-white solid.

[0769] 11H NMR (400 MHz, DMSO-d6) δ 9.12 (d, J = 8.31 Hz, 1H), 8.22 (d, J = 8.80 Hz, 1H), 8.10 (d, J = 8.80 Hz, 1H), 7.77 (d, J = 8.80 Hz, 1H), 7.27 (d, J = 8.80 Hz, 1H), 4.43 - 4.57 (m, 1H), 3.85 - 4.01 (m, 1H), 2.23 (s, 3H), 2.08 - 2.17 (m, 2H), 1.86 - 1.96 (m, 2H), 1.64 - 1.77 (m, 2H), 1.49 - 1.62 (m, 2H). LCMS: 404.9 [M+H] + 。

[0770] Step E4: Preparation of Ethyl 1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)-pyridazin-3-yl)piperidine-4-carboxylate (Intermediate E4)

[0771] At room temperature, potassium carbonate (5.86 g, 42 mmol, 2.5 equiv) was added to a stirred solution of 6-chloro-N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-pyridazine-3-carboxamide (Intermediate E3, 7 g, 17 mmol, 1.0 equiv) and ethyl piperidine-4-carboxylate (SM-4, 4 g, 25 mmol, 1.5 equiv) in DMF (70 mL). The reaction mixture was heated to 80 °C for 12 h. The progress of the reaction was monitored by TLC. After completion of the starting materials, the reaction mixture was diluted with ice water (300 mL), stirred for 10 minutes, and the precipitated solid was filtered and dried to give Intermediate E4 as an off-white solid (6 g, 66%).

[0772] 1 1H NMR (400 MHz, DMSO-d6) δ 8.57 (d, J = 7.82 Hz, 1H), 7.80 (d, J = 9.29 Hz, 1H), 7.75 (d, J = 8.80 Hz, 1H), 7.35 (d, J = 9.78 Hz, 1H), 7.24 (d, J = 8.80 Hz, 1H), 4.44 - 4.55 (m, 1H), 4.30 - 4.41 (m, 2H), 4.00 - 4.11 (m, 2H), 3.80 - 3.92 (m, 1H), 3.08 - 3.23 (m, 2H), 2.64 - 2.76 (m, 1H), 2.22 (s, 3H), 2.05 - 2.15 (m, 2H), 1.85 - 1.97 (m, 4H), 1.44 - 1.71 (m, 6H), 1.17 (t, J = 7.34 Hz, 3H). LCMS: 526.2 [M+H] +。

[0773] Step E5: Preparation of 1-(6-(((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carboxylic acid (Intermediate E)

[0774] At 0 °C, LiOH·H2O (2.49 g, 57.9 mmol, 5.0 eq) was added to a solution of ethyl 1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carboxylate (Intermediate E4, 6.0 g, 11.5 mmol, 1.0 eq) in THF (20 mL) and water (5 mL), and the reaction mixture was stirred for 5 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the volatiles were removed under reduced pressure, diluted with water (80 mL), acidified to pH 3 with 1N aqueous HCl and extracted with ethyl acetate (3 x 80 mL). The combined organic extracts were washed with water (200 mL), brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give Intermediate E as an off-white solid (5.4 g, 94%).

[0775] LCMS: 498.37 [M+H] + 。

[0776] Preparation of (2R,5S)-4-(1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carbonyl)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl ester formate

[0777] At 0 °C, HATU (1.14 g, 3.018 mmol, 1.5 eq) and DIPEA (1.75 mL, 10.06 mmol, 5.0 eq) were added to a stirred solution of (2S,5R)-2,5-dimethylpiperazine-1-carboxylic acid (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl ester (Intermediate D, 1.19 g, 2.012 mmol, 1.0 eq) and 1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)-pyridazin-3-yl)piperidine-4-carboxylic acid (Intermediate E, 1 g, 2.012 mmol, 1.0 eq) in DMF (10 mL), and the reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water (60 mL) and extracted with 10% methanol / DCM (2 × 60 mL). The combined organic extracts were washed with water (80 mL), brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product, which was purified by preparative HPLC (0.5% aqueous formic acid / CH3CN) to give the title compound as an off-white solid (600 mg, 28%).

[0778] 1 H NMR (400 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.59 (d, J = 8.13 Hz, 1H), 8.14 (dd, J = 8.94, 2.56 Hz, 1H), 7.81 - 7.85 (m, 1H), 7.77 (d, J = 8.76 Hz, 1H), 7.60 - 7.72 (m, 1H), 7.38 (dd, J = 9.51, 3.38 Hz, 1H), 7.35 (s, 1H), 7.26 (d, J = 8.88 Hz, 1H), 6.52 (s, 1H), 5.41 - 5.46 (m, 2H), 5.28 - 5.35 (m, 2H), 4.40 - 4.65 (m, 4H), 3.72 - 4.04 (m, 6H), 2.98 - 3.22 (m, 4H), 2.07 - 2.36 (m, 12H), 1.81 - 1.98 (m, 3H), 1.38 - 1.78 (m, 10H), 1.22 - 1.34 (m, 4H), 1.08 - 1.16 (m, 2H), 0.89 (t, J = 7.25 Hz, 3H). LCMS: 1041.4 [M+H] + . HPLC purity 98.6%.

[0779] Example S22: Preparation of methyl (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-((1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)(methyl)amino)hexyl)piperazine-1-carboxylate formate (Compound No. 22)

[0780]

[0781] Step F1: Preparation of tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate (Intermediate F1)

[0782] To a stirred solution of tert-butyl piperazine-1-carboxylate (SM-1, 5 g, 26.8 mmol, 1.0 equiv) in DCM (100 mL) at 0 °C was added pyridine (2.97 g, 37.6 mmol, 1.4 equiv) and triphosgene (3.19 g, 10.7 mmol, 0.4 equiv). The mixture was warmed to room temperature and stirred for 2 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the mixture was diluted with water (100 mL) and extracted with DCM (2 × 100 mL). The combined organic extracts were washed again with water (200 mL) and brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give Intermediate F1 as an oil (6.0 g, 90%).

[0783] 1 H NMR (400 MHz, DMSO-d6) δ 3.64 (d, J = 4.40 Hz, 2H), 3.52 (br s, 2H), 3.27 - 3.47 (m, 3H), 2.93 - 3.21 (m, 1H), 1.41 (s, 9H).

[0784] Step F2: Preparation of (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate · 4-(10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl) ester (Intermediate F2)

[0785] At 0 °C, DIPEA (15.3 g, 118 mmol, 5 eq) and DMAP (724 mg, 5.9 mmol, 0.25 eq) were added to a stirred solution of (S)-10-((dimethylamino)methyl)-4-ethyl-4,9-dihydroxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione hydrochloride (SM-2, 10 g, 23.7 mmol, 1.0 eq) in DCM (250 mL). Subsequently, tert-butyl 4-(chlorocarbonyl)piperazine-1-carboxylate (Intermediate F1, 5.89 g, 23.7 mmol, 1 eq) in DCM (20 mL) was added dropwise over 10 min, and the mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the mixture was diluted with water (100 mL) and extracted with DCM (3 × 100 mL). The combined organic extracts were washed again with water (200 mL) and brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product, which was purified by combi flash column using 7% methanol / DCM to give Intermediate F2 as an off-white solid (10 g, 66%).

[0786] 1 H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.11 (d, J = 9.29 Hz, 1H), 7.65 (d, J = 9.29 Hz, 1H), 7.34 (s, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 3.67 - 3.79 (m, 4H), 3.41 - 3.55 (m, 6H), 2.20 (s, 6H), 1.80 - 1.93 (m, 2H), 1.44 (s, 9H), 0.89 (t, J = 7.34 Hz, 3H). LCMS: 634.2 [M+H] + 。

[0787] Step F3: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl piperazine-1-carboxylate hydrochloride (Intermediate F)

[0788] Under a nitrogen atmosphere at 0 °C, 4 M HCl / 1,4-dioxane (50 mL) was added to (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate · 4-(10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-9-yl) ester (Intermediate F2, 5 g, 7.89 mmol, 1.0 equivalent). It was warmed to room temperature and stirred for 3 h. The reaction progress was monitored by TLC. After completion of the reaction, the volatiles were evaporated under reduced pressure, and the residue was triturated with diethyl ether (3 × 100 mL) to give Intermediate F (5 g, crude) as an off-white solid.

[0789] LCMS: 534.2 [M+H] + 。

[0790] Step 1: Preparation of tert-butyl (1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)(methyl)carbamate (Intermediate 1)

[0791] At room temperature, potassium carbonate (4.10 g, 29.7 mmol, 2.0 equivalents) was added to a stirred solution of 6-chloro-N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-pyridazine-3-carboxamide (Intermediate E3, 6 g, 14.85 mmol, 1.0 equivalent) and tert-butyl methyl(piperidin-4-yl)carbamate (SM-1, 4.76 g, 22.27 mmol, 1.5 equivalents) in DMF (60 mL). Subsequently, the reaction mixture was heated to 80 °C and stirred for 4 h. The reaction progress was monitored by TLC. After completion of the reaction, the mixture was diluted with ice water (300 mL), stirred for 10 minutes, and the precipitated solid was filtered and dried to give Intermediate 1 (6 g, 27%) as an off-white solid.

[0792] 11H NMR (400 MHz, DMSO-d6) δ 8.59 (d, J = 8.25 Hz, 1H), 7.82 (d, J = 9.63 Hz, 1H), 7.77 (d, J = 8.76 Hz, 1H), 7.38 (d, J = 9.63 Hz, 1H), 7.26 (d, J = 8.88 Hz, 1H), 4.56 - 4.64 (m, 2H), 4.46 - 4.55 (m, 1H), 3.97 - 4.23 (m, 1H), 3.81 - 3.94 (m, 1H), 2.95 - 3.07 (m, 2H), 2.64 (s, 3H), 2.24 (s, 3H), 2.08 - 2.15 (m, 2H), 1.86 - 1.95 (m, 2H), 1.49 - 1.71 (m, 8H), 1.39 (s, 9H). LCMS: 583.1 [M+H] + 。

[0793] Step 2: Preparation of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(methylamino)piperidin-1-yl)pyridazine-3-carboxamide hydrochloride (Intermediate 2)

[0794] To a solution of tert-butyl (1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)(methyl)carbamate (Intermediate 1, 7.0 g, 12.02 mmol, 1.0 equiv) at 0 °C was added 4 M HCl / 1,4-dioxane (70 mL), and the mixture was stirred at room temperature for 16 h. After completion of the reaction, the volatiles were evaporated under reduced pressure, and the resulting residue was washed with diethyl ether (2 × 80 mL) to give Intermediate 2 (7 g, crude) as an off-white solid, which was used without further purification.

[0795] 1 1H NMR (400 MHz, DMSO-d6) δ 9.13 (br s, 2H), 8.61 (d, J = 8.31 Hz, 1H), 7.88 (d, J = 9.78 Hz, 1H), 7.77 (d, J = 8.31 Hz, 1H), 7.47 (d, J = 9.78 Hz, 1H), 7.26 (d, J = 8.80 Hz, 1H), 4.45 - 4.56 (m, 2H), 3.83 - 3.96 (m, 1H), 3.57 (s, 3H), 3.08 (t, J = 12.23 Hz, 2H), 2.54 (t, J = 5.14 Hz, 2H), 2.24 (s, 3H), 2.06 - 2.18 (m, 4H), 1.86 - 1.96 (m, 2H), 1.48 - 1.72 (m, 6H). LCMS: 483.41 [M+H] +。

[0796] Step 3: Preparation of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-((6-hydroxyhexyl)-(methyl)amino)piperidin-1-yl)pyridazine-3-carboxamide (Intermediate 3)

[0797] At room temperature, NaHCO3 (6.1 g, 72.61 mmol, 10 eq) was added to a solution of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(methylamino)-piperidin-1-yl)pyridazine-3-carboxamide hydrochloride (Intermediate 2, 3.5 g, 7.26 mmol, 1.0 eq) and 6-bromohexan-1-ol (SM-2, 4.7 mL, 36.3 mmol, 5 eq) in ethanol (70 mL) and water (70 mL). The reaction mixture was heated to 100 °C and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the mixture was filtered through a pad of diatomaceous earth and washed with ethyl acetate (40 mL). The filtrate was concentrated under reduced pressure, diluted with water (120 mL) and extracted with ethyl acetate (2 × 200 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product, which was purified by combi flash chromatography eluting with 2-5% methanol / DCM to give Intermediate 3 as an off-white solid (4 g, 47%).

[0798] 1 1H NMR (400 MHz, DMSO-d6) δ 8.59 (d, J = 8.31 Hz, 1H), 7.82 (d, J = 9.29 Hz, 1H), 7.77 (d, J = 8.80 Hz, 1H), 7.37 (d, J = 9.29 Hz, 1H), 7.26 (d, J = 8.80 Hz, 1H), 4.46 - 4.62 (m, 3H), 4.29 - 4.35 (m, 1H), 3.83 - 3.94 (m, 1H), 3.34 - 3.41 (m, 3H), 3.00 (t, J = 12.23 Hz, 2H), 2.23 - 2.25 (m, 4H), 2.07 - 2.16 (m, 3H), 1.74 - 1.96 (m, 4H), 1.52 - 1.71 (m, 5H), 1.35 - 1.51 (m, 6H), 1.20 - 1.33 (m, 6H). LCMS: 583.17 [M+H] + 。

[0799] Step 4: Preparation of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(methyl(6-oxohexyl)amino)piperidin-1-yl)pyridazine-3-carboxamide (Intermediate 4)

[0800] At 0 °C, pyridinium chlorochromate (PCC, 2.21 g, 10.30 mmol, 1.5 eq) was added portionwise to a stirred solution of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-((6-hydroxyhexyl)(methyl)amino)piperidin-1-yl)pyridazine-3-carboxamide (Intermediate 3, 4.0 g, 6.87 mmol, 1 eq) in dichloromethane (50 mL), and the reaction mixture was stirred at room temperature for 4 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through a pad of diatomaceous earth, and the filtrate was concentrated under vacuum to give Intermediate 4 (4.2 g, crude) as a brown solid, which was used as such in the next step.

[0801] LCMS: 582.2 [M+H] + 。

[0802] Step 5: Preparation of (S)-10-((dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl 4-(6-((1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)(methyl)amino)hexyl)piperazine-1-carboxylate formate

[0803] (S)-10-((Dimethylamino)methyl)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-9-yl piperazine-1-carboxylate hydrochloride (Intermediate F, 3.67 gmg, 6.89 mmol, 1 equivalent) was added to a solution of methanol and dichloromethane (40, 160 mL), and Et3N (0.9 mL, 6.89 mmol, 1.0 equivalent) was added, and the resulting solution was stirred for 30 minutes. N-((1r,4r)-4-(3-Chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(methyl(6-oxohexyl)amino)piperidin-1-yl)pyridazine-3-carboxamide (Intermediate 4, 4 g, 6.89 mmol, 1 equivalent) and glacial acetic acid (0.1 mL) were added to this solution at room temperature, and the mixture was stirred for 2 h. NaBH(OAc)3 (5.81 mg, 27.58 mmol, 4 equivalents) was added to the reaction mixture at 0 °C. The reaction mixture was warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the volatiles were evaporated under reduced pressure, quenched with ice water (100 mL), and extracted with 10% methanol / DCM (2 × 80 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product, which was purified by preparative HPLC using mobile phase A: 0.1% HCOOH / water and mobile phase B: acetonitrile to give the title compound as an off-white solid (260 mg).

[0804] 11H NMR (400 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.57 (d, J = 7.83 Hz, 1H), 8.08 - 8.19 (m, 3H), 7.81 (d, J = 9.29 Hz, 1H), 7.76 (d, J = 8.31 Hz, 1H), 7.63 (d, J = 8.80 Hz, 1H), 7.32 - 7.40 (m, 2H), 7.21 - 7.28 (m, 1H), 6.51 (brs, 1H), 5.39 - 5.45 (m, 2H), 5.27 - 5.35 (m, 2H), 4.45 - 4.60 (m, 3H), 3.81 - 3.94 (m, 2H), 3.61 - 3.80 (m, 4H), 3.44 - 3.58 (m, 4H), 2.92 - 3.06 (m, 3H), 2.72 - 2.84 (m, 1H), 2.29 - 2.47 (m, 6H), 2.17 - 2.26 (m, 10H), 2.06 - 2.15 (m, 2H), 1.76 - 1.96 (m, 6H), 1.38 - 1.69 (m, 10H), 1.23 - 1.35 (m, 5H), 0.89 (t, J = 6.11 Hz, 3H). LCMS: 1098.0 [M+H] + . HPLC purity 95.6%.

[0805] Example S23: Preparation of (S)-4-Ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-4-yl 4-((1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)methyl)piperazine-1-carboxylate (Compound No. 23)

[0806]

[0807] Step 1: Preparation of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(hydroxymethyl)piperidin-1-yl)pyridazine-3-carboxamide (Intermediate 1)

[0808] To a flask was added 6-chloro-N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)pyridazine-3-carboxamide (Intermediate E3, 1 g, 2.46 mmol, 1.0 equiv), piperidin-4-ylmethanol (SM-1, 425 mg, 3.70 mmol, 1.5 equiv), K2CO3 (1 g, 7.40 mmol, 3.0 equiv) and DMF (10 mL, 10 vol). The reaction mixture was stirred at ambient temperature under a nitrogen atmosphere until TLC indicated complete consumption of the starting material. The reaction mixture was diluted with water (100 mL), extracted with ethyl acetate (2 × 200 mL), and the combined organic layers were washed with a brine solution (100 mL) and dried over sodium sulfate. The organic layer was filtered, concentrated under reduced pressure and purified by flash column (silica gel, 50 - 60% ethyl acetate / hexane). The pure fractions were combined and concentrated under reduced pressure to afford N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(hydroxymethyl)piperidin-1-yl)pyridazine-3-carboxamide (Intermediate 1, 1 g, 84%) as an off-white solid.

[0809] 1 H NMR (400 MHz, DMSO-d6) δ 8.58 (d, J = 8.31 Hz, 1H), 7.95 (s, 1H), 7.78 (t, J = 9.05 Hz, 2H), 7.33 (d, J = 9.78 Hz, 1H), 7.26 (d, J = 8.80 Hz, 1H), 4.42 - 4.55 (m, 5H), 3.81 - 3.94 (m, 1H), 3.27 (t, J = 5.14 Hz, 2H), 2.23 (s, 3H), 2.11 (d, J = 10.76 Hz, 2H), 1.90 (d, J = 10.27 Hz, 2H), 1.46 - 1.81 (m, 7H), 1.06 - 1.22 (m, 2H). LCMS: 484.4 [M+H] + 。

[0810] Step 2: Preparation of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-formylpiperidin-1-yl)pyridazine-3-carboxamide (Intermediate 2)

[0811] Add N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(hydroxymethyl)piperidin-1-yl)pyridazine-3-carboxamide (Intermediate 1, 800 mg, 1.65 mmol, 1.0 eq), Dess-Martin periodinane (1 g, 2.47 mmol, 1.5 eq) and DCM (10 mL, 10 vol) to a flask. Stir the reaction mixture under a nitrogen atmosphere at ambient temperature until TLC indicates complete consumption of the starting material. Quench the reaction mixture with saturated bicarbonate solution (100 mL), and extract the desired compound with DCM (2 x 200 mL). Wash the combined organic layers with brine solution (100 mL) and dry over sodium sulfate. Filter the organic solvent and concentrate it under reduced pressure to obtain N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-formylpiperidin-1-yl)pyridazine-3-carboxamide (Intermediate 2, 750 mg, 94%) as an off-white solid, which can be used without further purification.

[0812] 1 H NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 8.60 (d, J = 7.83 Hz, 1H), 7.8 (d, J = 8.40 Hz, 1H), 7.37 (d, J = 9.78 Hz, 1H), 7.26 (d, J = 8.80 Hz, 1H), 4.48–4.50 (m, 1H), 4.31 (d, J = 13.21 Hz, 2H), 3.86 - 3.89 (m, 1H), 3.26 (t, J = 11.98 Hz, 2H), 2.61 - 2.76 (m, 1H), 2.20 - 2.30 (m, 4H), 2.10–2.12 (m, 2H), 1.81 - 2.00 (m, 4H), 1.46 - 1.70 (m, 6H). LCMS: 482.4 [M+H] + 。

[0813] Step 3: Preparation of tert-butyl 4-((1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)methyl)piperazine-1-carboxylate (Intermediate 3)

[0814] At room temperature, catalytic amount of glacial acetic acid (0.4 mL) was added to a stirred solution of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-formylpiperidin-1-yl)pyridazine-3-carboxamide (Intermediate 2, 1.8 g, 3.74 mmol, 1 equiv) and tert-butyl piperazine-1-carboxylate (SM-2, 1.38 g, 7.46 mmol, 2 equiv) in methanol (18 mL) and stirred for 3 h. At 0 °C, NaBH3CN (462 mg, 7.46 mmol, 2 equiv) was added to the reaction mixture. The reaction mixture was warmed to room temperature and stirred for 16 h. The reaction progress was monitored by TLC and LCMS. After completion of the reaction, the solvent was evaporated under reduced pressure, quenched with ice water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give the crude product. The obtained crude product was purified by combiflash column chromatography eluting with 80% EtOAc / heptane to give Intermediate 3 (1.0 g, 41%) as an off-white solid.

[0815] 1 H NMR (400 MHz, DMSO-d6) δ 8.58 (d, J = 7.34 Hz, 1H), 7.78 (t, J = 7.83 Hz, 2H), 7.32 (d, J = 8.31 Hz, 1H), 7.26 (d, J = 8.31 Hz, 1H), 4.42 - 4.57 (m, 3H), 3.81 - 3.94 (m, 1H), 3.27 - 3.31 (m, 4H), 2.99 (t, J = 12.23 Hz, 2H), 2.26 - 2.32 (m, 4H), 2.24 (s, 3H), 2.07 - 2.18 (m, 4H), 1.75 - 1.96 (m, 5H), 1.48 - 1.72 (m, 4H), 1.39 (s, 9H), 1.02 - 1.19 (m, 2H). LCMS: 652.64 [M+H] + 。

[0816] Step 4: Preparation of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(piperazin-1-ylmethyl)piperidin-1-yl)pyridazine-3-carboxamide trifluoroacetate (Intermediate 4)

[0817] Under a nitrogen atmosphere at 0 °C, trifluoroacetic acid (TFA, 1.17 mL) was added to a stirred solution of tert-butyl 4-((1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)methyl)piperazine-1-carboxylate (Intermediate 3, 500 mg, 0.77 mmol, 1.0 equiv.) in DCM (10 mL), and the reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure to give the crude product. The obtained crude product was triturated twice with n-pentane (2 x 30 mL) to give N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(piperazin-1-ylmethyl)piperidin-1-yl)pyridazine-3-carboxamide trifluoroacetate (Intermediate 4, 348 mg, 82%) as an off-white solid.

[0818] LCMS: 552.68 [M+H] + 。

[0819] Step 5: Preparation of (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-4-yl 4-((1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidin-4-yl)methyl)piperazine-1-carboxylate

[0820] To a solution of (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-4-yl 4-nitrobenzoate (Intermediate 4, 600 mg, 1.1 mmol, 1 equiv.) and N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(piperazin-1-ylmethyl)piperidin-1-yl)pyridazine-3-carboxamide trifluoroacetate (Intermediate 5, 730 mg, 1.32 mmol, 1.2 equiv.) in acetonitrile (10 mL) was added K2CO3 (457 mg, 3.31 mmol, 3 equiv.). The resulting reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by LCMS / TLC. After completion of the reaction, the mixture was diluted with water (40 mL) and extracted with ethyl acetate (2 x 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC using mobile phase A: 0.1% FA / water and mobile phase B: acetonitrile to give the title compound (95 mg, 8%) as an off-white solid.

[0821] 1 1H NMR (400 MHz, DMSO-d6) δ 8.55 - 8.59 (m, 2H), 8.06 (d, J = 9.01 Hz, 1H), 7.80 (d, J = 9.63 Hz, 1H), 7.77 (d, J = 8.88 Hz, 1H), 7.49 - 7.55 (m, 2H), 7.33 (d, J = 9.76 Hz, 1H), 7.26 (d, J = 8.88 Hz, 1H), 7.00 (s, 1H), 5.44 (d, J = 3.88 Hz, 2H), 5.29 (s, 2H), 4.42 - 4.57 (m, 3H), 3.95 (s, 3H), 3.83 - 3.93 (m, 1H), 3.67 - 3.78 (m, 1H), 3.55 - 3.65 (m, 1H), 3.18 - 3.26 (m, 2H), 2.94 - 3.06 (m, 2H), 2.53 - 2.58 (m, 1H), 2.29 - 2.37 (m, 1H), 2.24 (s, 3H), 2.07 - 2.22 (m, 8H), 1.86 - 1.96 (m, 3H), 1.75 - 1.85 (m, 2H), 1.48 - 1.71 (m, 4H), 1.05 - 1.19 (m, 2H), 0.91 (t, J = 7.44 Hz, 3H). LCMS: 956.2 [M+H] + . HPLC purity 95.7%.

[0822] Example S24: Preparation of (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-4-carboxylate formate of 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylic acid (Compound No. 24)

[0823]

[0824] Step 1: Preparation of (S)-4-ethyl-4-hydroxy-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione (Intermediate 1)

[0825] At 0 °C, K2CO3 (752 mg, 5.49 mmol, 2 equiv) was added to a stirred solution of 10-hydroxycamptothecin (SM-1, 1 g, 2.75 mmol, 1 equiv) in DMF (10 mL), followed by the addition of methyl iodide (0.25 mL, 4.12 mmol, 1.5 equiv). The reaction mixture was stirred at 80 °C for 3 h. The reaction progress was monitored by TLC / LCMS. After completion of the reaction, the mixture was poured into ice water (30 mL) and the resulting solid was filtered, washed with cold water and dried to give (S)-4-ethyl-4-hydroxy-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione (Intermediate 1, 0.93 g, 90%) as an off-white solid.

[0826] 1 1H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.06 (d, J = 9.78 Hz, 1H), 7.47 - 7.53 (m, 2H), 7.28 (s, 1H), 6.49 (s, 1H), 5.41 (s, 2H), 5.25 (s, 2H), 3.94 (s, 3H), 1.78 - 1.95 (m, 2H), 0.88 (t, J = 7.34 Hz, 3H). LCMS: 379.36 [M+H] + 。

[0827] Step 2: Preparation of (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-4-yl carbonate · (4-nitrophenyl) ester (Intermediate 2)

[0828] At 0 °C, to a stirred solution of (S)-4-ethyl-4-hydroxy-9-methoxy-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione (Intermediate 1, 1.5 g, 3.96 mmol, 1 equiv) in DCM (15 mL) was added 4-nitrophenyl chloroformate (1.19 g, 5.9 mmol, 1.5 equiv), DMAP (98 mg, 0.79 mmol, 0.2 equiv) and triethylamine (1.10 mL, 7.9 mmol, 2 equiv). The reaction mixture was stirred at room temperature for 4 h. The reaction progress was monitored by TLC and LCMS. After completion of the reaction, the mixture was quenched with ice water (20 mL) and extracted with 10% MeOH / DCM (2 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained crude product was triturated with diethyl ether twice (2 x 20 mL) to give (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-4-yl carbonate·(4-nitrophenyl) ester (Intermediate 2, 2.0 g, crude) as a pale yellow solid.

[0829] 1 H NMR (400 MHz, DMSO-d6) δ 8.49 (d, J = 6.85 Hz, 1H), 8.27 (t, J = 8.80 Hz, 1H), 8.08 (d, J = 8.80 Hz, 2H), 8.01 - 8.04 (m, 1H), 7.44 - 7.50 (m, 2H), 6.86 (d, J = 8.80 Hz, 2H), 5.38 - 5.51 (m, 2H), 5.18 - 5.26 (m, 2H), 3.92 (s, 3H), 2.10 - 2.32 (m, 2H), 0.85 - 0.95 (m, 3H).

[0830] Step 3: Preparation of (S)-1-(tert-butyl) 4-(4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-4-yl) piperazine-1,4-dicarboxylate (Intermediate 3)

[0831] To a stirred solution of (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-4-carboxylic acid (4-nitrophenyl) ester (Intermediate 2, 200 mg, 0.36 mmol, 1 equiv) in acetonitrile (5 mL) was added tert-butyl piperazine-1-carboxylate (82 mg, 0.44 mmol, 1.2 equiv) and K2CO3 (149 mg, 1.08 mmol, 3 equiv). The reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the mixture was diluted with water (20 mL) and extracted with 5% MeOH / DCM (2 x 20 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude product was purified by combiflash column chromatography eluting with 100% ethyl acetate to afford (S)-1-(tert-butyl) piperazine-1,4-dicarboxylate 4-(4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-4-yl) ester (Intermediate 3, 110 mg, 50%) as an off-white solid.

[0832] 1 H NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1H), 8.05 (d, J = 9.29 Hz, 1H), 7.46 - 7.56 (m, 2H), 7.01 (s, 1H), 5.42 - 5.47 (m, 2H), 5.28 (s, 2H), 3.94 (s, 3H), 3.58 - 3.74 (m, 2H), 3.37 - 3.55 (m, 2H), 3.21 - 3.28 (m, 4H), 2.10 - 2.20 (m, 2H), 1.41 (s, 9H), 0.91 (t, J = 7.34 Hz, 3H). LCMS: 591.58 [M+H] + 。

[0833] Step 4: Preparation of (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-4-yl piperazine-1-carboxylate (Intermediate 4)

[0834] Under a nitrogen atmosphere, at 0 °C, trifluoroacetic acid (TFA, 1.6 mL) was added to a stirred solution of piperazine-1,4-dicarboxylic acid (S)-1-(tert-butyl) ester · 4-(4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-4-yl) ester (Intermediate 3, 480 mg, 0.81 mmol, 1.0 equiv) in DCM (5 mL), and the reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, saturated NaHCO3 solution (20 mL) was added, and the aqueous solution was extracted with DCM (2 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was triturated twice with n-pentane (2 x 30 mL) to give piperazine-1-carboxylic acid (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-4-yl ester (Intermediate 4, 380 mg, 95%) as an off-white solid.

[0835] 1 H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.06 (d, J = 8.80 Hz, 1H), 7.46 - 7.54 (m, 2H), 6.97 (s, 1H), 5.36 - 5.48 (m, 2H), 5.26 (s, 2H), 3.93 (s, 3H), 3.50 - 3.63 (m, 2H), 3.09 - 3.22 (m, 2H), 2.70 - 2.89 (m, 2H), 2.53 - 2.66 (m, 2H), 2.07 - 2.16 (m, 2H), 0.88 (t, J = 7.09 Hz, 3H). No exchangeable protons were observed. LCMS: 491.49 [M+H] + 。

[0836] Step 5: Preparation of (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-4-yl 4-(6-((4-((8S,11R,13S,14S,17R)-17-acetoxy-17-acetyl-13-methyl-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-11-yl)phenyl)(methyl)amino)hexyl)piperazine-1-carboxylate formate

[0837] At room temperature, triethylamine (0.14 mL) and glacial acetic acid (0.06 mL) were added to a stirred solution of (8S,11R,13S,14S,17R)-17-acetyl-13-methyl-11-(4-(methyl(6-oxohexyl)amino)phenyl)-3-oxo-2,3,6,7,8,11,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl acetate (Intermediate C, 855 mg, 1.53 mmol, 1.5 equiv) and (S)-4-ethyl-9-methoxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]-indazino[1,2-b]quinolin-4-yl piperazine-1-carboxylate (Intermediate 4, 500 mg, 1.02 mmol, 1 equiv) in methanol (10 mL), and the mixture was stirred for 2 h. NaCNBH3 (126 mg, 2.04 mmol, 2 equiv) was added to the reaction mixture at 0 °C, warmed to room temperature and stirred for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated under reduced pressure, quenched with ice water (20 mL) and extracted with 10% methanol / DCM (2 x 20 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product obtained was purified by preparative HPLC eluting with mobile phase A: 0.1% FA / water and mobile phase B: acetonitrile to give the title compound as an off-white solid (90 mg, 8%).

[0838] 1 H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.04 (d, J = 9.13 Hz, 1H), 7.44 - 7.56 (m, 2H), 6.93 - 7.02 (m, 3H), 6.58 (d, J = 8.76 Hz, 2H), 5.66 (s, 1H), 5.37 - 5.49 (m, 2H), 5.28 (s, 2H), 4.39 (d, J = 6.75 Hz, 1H), 3.94 (s, 3H), 3.49 - 3.81 (m, 2H), 3.16 - 3.25 (m, 4H), 2.81 (s, 3H), 2.64 - 2.76 (m, 2H), 2.52 - 2.61 (m, 4H), 2.42 - 2.47 (m, 2H), 2.21 - 2.38 (m, 5H), 2.11 - 2.19 (m, 5H), 2.09 (s, 3H), 1.99 (s, 2H), 1.85 - 1.95 (m, 2H), 1.61 - 1.77 (m, 3H), 1.18 - 1.53 (m, 12H), 0.91 (t, J = 7.32 Hz, 3H), 0.23 (s, 3H). LCMS: 1034.4 [M+H] +。The HPLC purity is 93.8%.

[0839] Example S25: Preparation of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-(4-(((S)-4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazine-1-carbonyl)piperidin-1-yl)pyridazine-3-carboxamide (Compound No. 25)

[0840]

[0841] To a stirred solution of 1-(6-(((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)carbamoyl)pyridazin-3-yl)piperidine-4-carboxylic acid (Intermediate E, 4 g, 8.26 mmol, 1 equiv.) in DMF (40 mL) was added EDC·HCl (2.37 g, 12.39 mmol, 1.5 equiv.), HOBT (1.89 g, 12.39 mmol, 1.5 equiv.) and DIPEA (3.19 g, 24.78 mmol, 3.0 equiv.). At 0 °C, (S)-4-ethyl-4,9-dihydroxy-10-(piperazin-1-ylmethyl)-1,12-dihydro-14H-pyrano[3',4':6,7]indazino[1,2-b]quinoline-3,14(4H)-dione trifluoroacetate (Intermediate B, 5.7 g, 9.91 mmol, 1.2 equiv.) was added and the reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with ice water (120 mL), and the formed solid was filtered and dried to give 7.6 g of a crude product as a pale yellow solid. A portion of the crude product (2 g) was purified by preparative HPLC using mobile phase A: 0.1% FA / water and mobile phase B: acetonitrile to give the title compound (508 mg) as a pale yellow solid.

[0842] 11H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1H), 8.59 (d, J = 7.82 Hz, 1H), 8.00 (d, J = 9.29 Hz, 1H), 7.74 - 7.85 (m, 2H), 7.48 (d, J = 8.80 Hz, 1H), 7.36 (d, J = 9.29 Hz, 1H), 7.24 - 7.29 (m, 2H), 6.49 (s, 1H), 5.37 - 5.45 (m, 2H), 5.23 - 5.30 (m, 2H), 4.43 - 4.57 (m, 3H), 4.04 (s, 2H), 3.83 - 3.95 (m, 1H), 3.43 - 3.63 (m, 5H), 2.97 - 3.18 (m, 4H), 2.54 - 2.64 (m, 2H), 2.24 (s, 3H), 2.07 - 2.17 (m, 2H), 1.80 - 1.96 (m, 4H), 1.48 - 1.78 (m, 9H), 0.88 (t, J = 7.09 Hz, 3H). LCMS: 942.3 [M+H] + . HPLC purity 96.5%.

[0843] Example S26: Preparation of N-((1r,4r)-4-(3-chloro-4-cyano-2-methylphenoxy)cyclohexyl)-6-(4-((4-(((S)-4-ethyl-4,9-dihydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indazino[1,2-b]quinolin-10-yl)methyl)piperazin-1-yl)methyl)piperidin-1-yl)pyridazine-3-carboxamide formate (Compound No. 26)

[0844]

[0845] At room temperature, trie...

Claims

1. A compound of formula I, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof: A 1 -L 1 -B 1 I Wherein: B 1 is a nuclear receptor targeting epitope; L 1 is a covalent bond or a linking moiety; and A 1 having the formula IA: Wherein: R 1 、R 2 、R 3 、R 4 and R 5 each independently is hydrogen, a halogen group, a cyano group, a nitro group, -OR 15 、-SR 15 、-NR 15 R 16 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl, a 5- to 12-membered heteroaryl, -C(=O)R 15 、-C(=O)OR 15 、-OC(=O)R 15 、-OC(=O)NR 15 R 16 、-C(=O)NR 15 R 16 、-NR 15 C(=O)R 16 、-NR 15 C(=O)OR 16 、-S(=O) 1-2 R 15 、-S(=O) 1-2 NR 15 R 16 、-NR 15 S(=O) 1-2 R 16 、-Si(R 15 )3 or -C=NOR 15 ,and each independently is optionally substituted by one or more R 10 when the valence allows; or R 1 and R 2 with R 1 and R 2 together with the attached atom form C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted by one or more R 10 substituted; or R 2 and R 3 with R 2 and R 3 together with the attached atom form a C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted by one or more R 10 substituents; or R 3 and R 4 with R 3 and R 4 together with the attached atom form a C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted by one or more R 10 substituents; Each R 10 is independently a halogen group, a cyano group, a nitro group, -OR 17 , -SR 17 , -SF5, -NR 17 R 18 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5-12 membered heterocyclic group, C 6-12 aryl, 5-12 membered heteroaryl, -C(=O)R 17 , -C(=O)OR 17 , -OC(=O)OR 17 , -OC(=O)R 17 , -C(=O)NR 17 R 18 , -OC(=O)NR 17 R 18 , -NR 7 C(=O)NR 17 R 18 , -S(=O) 1- 2R 17 , -S(=O) 1-2 NR 17 R 18 , -NR 17 S(=O) 1-2 R 18 , -NR 17 S(=O) 1-2 NR 17 R 18 , -NR 17 C(=O)R 18 , -NR 17 C(=O)OR 18 , -Si(R 17 )3 or -C=NOR 17 , which are each independently optionally substituted, when valence permits, by one or more substituents selected from the group consisting of: halogen group, cyano group, nitro group, hydroxy group, amino group, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5-12 membered heterocyclic group, C 6-12 aryl and 5-12 membered heteroaryl; Each R 15 and R 16 are each independently hydrogen, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocycloalkyl, C 6-12 aryl, or 5- to 12-membered heteroaryl, each of which is independently optionally substituted, where valency permits, with one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocycloalkyl, C 6-12 aryl, and 5- to 12-membered heteroaryl; or R 15 and R 16 with R 15 and R 16 together with the attached atom form a 5- to 12-membered heterocyclic group, which is optionally substituted, where valency permits, with one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl and 5- to 12-membered heteroaryl; and Each R 17 and R 18 independently is hydrogen, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocycloalkyl, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted, when valency permits, with one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocycloalkyl, C 6-12 aryl and 5- to 12-membered heteroaryl; or R 17 and R 18 with R 17 and R 18 together with the attached atom(s) form a 5- to 12-membered heterocyclic group which is optionally substituted, where valency permits, by one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl and 5- to 12-membered heteroaryl; wherein one or more atoms of formula IA (e.g., hydrogen, methyl or hydroxyl) are replaced by a direct covalent bond with L 1 is replaced.

2. The compound according to claim 1, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein the compound is not a compound selected from the group of compounds in Table 1X, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof.

3. The compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen.

4. The compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 1 is C 1-12 alkyl, which is optionally substituted by one or more R 10 substituents.

5. The compound according to claim 1 or 2, or a stereoisomer, a mixture of stereoisomers, a hydrate, a solvate, an isotope-enriched analogue or a pharmaceutically acceptable salt thereof, wherein R 1 is -Si(R 15 )3, which is optionally substituted by one or more R 10 substituents.

6. The compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 1 is -C=NOR 15 , which is optionally substituted by one or more R 10 .

7. A compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 1 is ethyl.

8. A compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 1 is 9. A compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 1 is 10. A compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 1 is 11. The compound according to claim 1 or 2, or a stereoisomer, a mixture of stereoisomers, a hydrate, a solvate, an isotope-enriched analogue or a pharmaceutically acceptable salt thereof, wherein R 1 is 12. A compound according to any one of claims 1-11, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 2 is hydrogen.

13. A compound according to any one of claims 1-11, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 2 is C 1-12 alkyl, which is optionally substituted by one or more R 10 substituents.

14. A compound according to any one of claims 1-11, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 2 is 15. A compound according to any one of claims 1-11, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 2 is nitro.

16. A compound according to any one of claims 1-11, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 2 is 17. A compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 1 and R 2 together with the atoms to which R 1 and R 2 are attached form a C 3-12 cycloalkyl which is optionally substituted by one or more R 10 substituents.

18. A compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 1 and R 2 together with the atoms to which R 1 and R 2 is attached form 19. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is -OR 15 , which is optionally substituted by one or more R 10 substituents.

20. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is C 1-12 alkyl, which is optionally substituted with one or more R 10 substituents.

21. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is -OC(=O)NR 15 R 16 , which is optionally substituted by one or more R 10 .

22. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is -OH.

23. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is methyl.

24. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is 25. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is 26. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is 27. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is 28. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is 29. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is a methoxy group.

30. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 is hydrogen.

31. A compound according to any one of claims 1 - 30, or a stereoisomer thereof, a mixture of stereoisomers, a hydrate, a solvate, an isotope - enriched analogue or a pharmaceutically acceptable salt, wherein R 4 is hydrogen.

32. A compound according to any one of claims 1-30, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 4 is a halogenated group.

33. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 and R 4 together with the atoms to which R 3 and R 4 are attached form a 5- to 12-membered heterocyclic group, which is optionally substituted with one or more R 10 substituents.

34. A compound according to any one of claims 1-18, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 3 and R 4 together with the atoms to which R 3 and R 4 is attached form 35. A compound according to any one of claims 1-34, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 5 is hydrogen.

36. A compound according to any one of claims 1-34, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 5 is -C(=O)R 15 , which is optionally substituted by one or more R 10 .

37. A compound according to any one of claims 1 - 34, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R 5 is 38. A compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein A 1 is derived from:

39. A compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein A 1 is derived from:

40. A compound according to any one of claims 1-39, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein the hydrogen atom of formula IA is replaced by a direct covalent bond with L 1 is replaced.

41. A compound according to any one of claims 1 - 39, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein the methyl group of formula IA is replaced by a direct covalent bond with L 1 is replaced.

42. A compound according to any one of claims 1 - 39, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein the hydroxyl group of formula IA is replaced by a direct covalent bond with L 1 is replaced.

43. A compound according to any one of claims 1-39, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein L 1 is linked to 1 the nitrogen atom of A.

44. A compound according to any one of claims 1-39, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein L 1 is attached to an oxygen atom of A 1 .

45. A compound according to claim 1 or 2, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein A 1 is:

46. The compound according to claim 1 or 2, or a stereoisomer thereof, a mixture of stereoisomers, a hydrate, a solvate, an isotope-enriched analogue or a pharmaceutically acceptable salt thereof, wherein A 1 is:

47. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 binds to the estrogen receptor.

48. A compound according to any one of claims 1 - 47, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 binds to the glucocorticoid receptor.

49. A compound according to any one of claims 1-48, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 binds to the progesterone receptor.

50. A compound according to any one of claims 1 - 49, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 binds to the androgen receptor.

51. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 has formula IIA: Wherein: The wavy key represents the connection point with L 1 ; R 30 is hydrogen, C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, C 2-12 alkynyl or C 3-12 cycloalkyl, where each C, when valency permits, 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, C 2-12 alkynyl or C 3-12 cycloalkyl is optionally and independently substituted by one or more R 100 substituents; R 40 is hydrogen, C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, C 2-12 alkynyl or C 3-12 cycloalkyl, where each C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl, C 2-12 alkynyl or C 3-12 cycloalkyl is optionally and independently substituted by one or more R 100 substituents; Each R 50 and R 51 independently is a halogen group, a cyano group, a nitro group, -OR 170 , -SR 170 , -NR 170 R 180 , C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl or C 2-12 alkynyl; provided that when the valence allows, each C 1-12 alkyl, C 1-12 haloalkyl, C 2-12 alkenyl or C 2-12 alkynyl is independently optionally substituted by one or more halogen groups, hydroxyl groups or amino groups; Each R 100 is independently an oxo group, a halogen group, a cyano group, a nitro group, -OR 170 、-SR 170 、-SF5, -NR 170 R 180 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl, a 5- to 12-membered heteroaryl, -C(=O)R 170 、-C(=O)OR 170 、-OC(=O)OR 170 、-OC(=O)R 170 、-C(=O)NR 170 R 180 、-OC(=O)NR 170 R 180 、-NR 170 C(=O)NR 170 R 180 、-S(=O) 1-2 R 170 、-S(=O) 1-2 NR 170 R 180 、-NR 170 S(=O) 1-2 R 180 、-NR 170 S(=O) 1-2 NR 170 R 180 、-NR 170 C(=O)R 180 or -NR 170 C(=O)OR 180 , each of which is independently optionally substituted, when valence permits, by one or more substituents selected from the group consisting of: a halogen group, a cyano group, a nitro group, a hydroxyl group, an amino group, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl and a 5- to 12-membered heteroaryl; and Each R 170 and R 180 is independently hydrogen or C 1-12 alkyl, which is optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group when valence permits, or R 170 and R 180 with R 170 and R 180 Together with the attached atom, form a heterocyclic group optionally substituted with a halogen group or a C optionally substituted with an oxo group, a halogen group, a hydroxyl group or an amino group 1-12 alkyl group.

52. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 53. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 has formula IIB': Wherein: The wavy key represents the connection point with L 1 ; R N is H or C 1-12 alkyl; R 60 is hydrogen, -OR 101 、-NR 101 R 102 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, 5- to 12-membered heteroaryl, -C(=O)R 101 、-C(=O)OR 101 、-OC(=O)R 101 、-OC(=O)NR 101 R 102 、-C(=O)NR 101 R 102 、-NR 101 C(=O)R 102 、-NR 101 C(=O)OR 102 , each of which is optionally and independently substituted by one or more R 100 substituents where valency permits; R 80 is hydrogen, -OR 101 、-NR 101 R 102 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, 5- to 12-membered heteroaryl, -C(=O)R 101 、-C(=O)OR 101 、-OC(=O)R 101 、-OC(=O)NR 101 R 102 、-C(=O)NR 101 R 102 、-NR 101 C(=O)R 102 、-NR 101 C(=O)OR 102 , each of which is optionally and independently substituted by one or more R 100 substituents when the valence allows; R 81 is hydrogen, -OR 101 、-NR 101 R 102 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl, 5- to 12-membered heteroaryl, -C(=O)R 101 、-C(=O)OR 101 、-OC(=O)R 101 、-OC(=O)NR 101 R 102 、-C(=O)NR 101 R 102 、-NR 101 C(=O)R 102 、-NR 101 C(=O)OR 102 which, where valency permits, are each independently optionally substituted by one or more R 100 substituents; or R 80 and R 81 with R 80 and R 81 together with the attached atom form a heterocyclic group optionally substituted by a halogen group or a C optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group 1-12 alkyl; R 82 is hydrogen, -OR 101 、-NR 101 R 102 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5 - 12 - membered heteroaryl, C 6-12 aryl, 5 - 12 - membered heteroaryl, -C(=O)R 101 、-C(=O)OR 101 、-OC(=O)R 101 、-OC(=O)NR 101 R 102 、-C(=O)NR 101 R 102 、-NR 101 C(=O)R 102 、-NR 101 C(=O)OR 102 and each of them is optionally and independently substituted by one or more R 100 substituents, when the valence allows. Each R 101 and R 102 independently is hydrogen, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl or 5- to 12-membered heteroaryl, each of which is independently optionally substituted, when valency permits, by one or more substituents selected from the group consisting of: halo, cyano, nitro, hydroxy, amino, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, 5- to 12-membered heterocyclic group, C 6-12 aryl and 5- to 12-membered heteroaryl; Each R 100 is independently an oxo group, a halogen group, a cyano group, a nitro group, -OR 170 , -SR 170 , -SF5, -NR 170 R 180 , C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl, a 5- to 12-membered heteroaryl, -C(=O)R 170 , -C(=O)OR 170 , -OC(=O)OR 170 , -OC(=O)R 170 , -C(=O)NR 170 R 180 , -OC(=O)NR 170 R 180 , -NR 170 C(=O)NR 170 R 180 , -S(=O) 1-2 R 170 , -S(=O) 1-2 NR 170 R 180 , -NR 170 S(=O) 1-2 R 180 , -NR 170 S(=O) 1-2 NR 170 R 180 , -NR 170 C(=O)R 180 or -NR 170 C(=O)OR 180 , which are each independently optionally substituted, where valency permits, by one or more substituents selected from the group consisting of: a halogen group, a cyano group, a nitro group, a hydroxy group, an amino group, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl and a 5- to 12-membered heteroaryl; and Each R 170 and R 180 is independently hydrogen or C 1-12 alkyl, which is optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group when valency permits, or R 170 and R 180 with R 170 and R 180 Together with the attached atom, form a heterocyclic group optionally substituted by a halogen group or a C 1-12 alkyl group optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group.

54. The compound according to claim 53, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein R N is methyl.

55. A compound according to any one of claims 1 - 46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 56. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 57. A compound according to any one of claims 1 - 46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 58. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 59. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 has formula IIC': Wherein: The wavy key represents the connection point with L 1 ; A” and A”' are each independently O or S; R a and R b are each independently CH3 or CH2CH3; or R a and R b together with the atom to which R a and R b is attached form a C 3-6 cycloalkyl, oxirane, oxetane or tetrahydrofuran; B, B 10 , B 2 , B 3 , B', B 1’ , B 2’ and B 3’ each independently is CR c or N; Each R c is independently hydrogen, fluorine, CN or methyl; D is absent, or is NH, O, S, CH2, -NH(C=O)-, -(C=O)NH- or C=O; X ” is CN, a halogenated group or NO2; Y ” is CH3, CH2R d , CHF2 or CF3; R d is a halogen; "Z" is H, C 1-2 alkyl, C2 alkenyl or NO2; or "X" and "Y" together form where the dash indicates the bond to the ring; or Y” and Z” together form where each is a single bond or a double bond, and where the dash indicates a bond to the ring; and Z' is CH or N.

60. The compound according to claim 59, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein D is NH, O, S, CH2, -NH(C=O)-, -(C=O)NH- or C=O.

61. A compound according to any one of claims 1 - 46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 62. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 63. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 has formula IID': Wherein: W is O, S or NH; Each is independently a double bond or a single bond; Each R 61 and R 62 is independently hydrogen, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl or C 3-12 cycloalkyl, where when valency permits, each C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl or C 3-12 cycloalkyl is optionally independently substituted by one or more R 100 substituents; Each R 100 is independently an oxo group, a halogen group, a cyano group, a nitro group, -OR 170 、-SR 170 、-SF5, -NR 170 R 180 、C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl, a 5- to 12-membered heteroaryl, -C(=O)R 170 、-C(=O)OR 170 、-OC(=O)OR 170 、-OC(=O)R 170 、-C(=O)NR 170 R 180 、-OC(=O)NR 170 R 180 、-NR 170 C(=O)NR 170 R 180 、-S(=O) 1-2 R 170 、-S(=O) 1-2 NR 170 R 180 、-NR 170 S(=O) 1-2 R 180 、-NR 170 S(=O) 1-2 NR 170 R 180 、-NR 170 C(=O)R 180 or -NR 170 C(=O)OR 180 , each of which is independently optionally substituted, when valency permits, by one or more substituents selected from the group consisting of: a halogen group, a cyano group, a nitro group, a hydroxyl group, an amino group, C 1-12 alkoxy, C 1-12 alkyl, C 2-12 alkenyl, C 2-12 alkynyl, C 3-12 cycloalkyl, a 5- to 12-membered heterocyclic group, C 6-12 aryl and a 5- to 12-membered heteroaryl; and Each R 170 and R 180 is independently hydrogen or C 1-12 alkyl, which is optionally substituted by an oxo group, a halogen group, a hydroxyl group or an amino group when valence permits, or R 170 and R 180 with R 170 and R 180 Together with the attached atom, form a heterocyclic group optionally substituted with a halogen group or a C 1-12 alkyl group optionally substituted with an oxo group, a halogen group, a hydroxyl group or an amino group.

64. The compound according to claim 63, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 has the formula IID:

65. A compound according to any one of claims 1 - 46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 66. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 67. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 has formula IIE: Wherein: The wavy bond refers to the point of attachment to L; Q is wherein key a is attached to loop a and key b is attached to loop b; R a and R b each independently is -CH3 or -CH2CH3; or R a and R b together with the atom to which R a and R b is attached form a C 3-5 cycloalkyl, oxiranyl, oxetanyl or tetrahydrofuranyl; A and A' are each independently O or S; E, E 1 , E 2 and E 3 are each independently CR c or N, and each R c is independently hydrogen, a halogen group, CN or methyl; E 4 is CF, CH or N; Q 1 is a bond, CH2, C=O or (C=O)NH; Q 2 is NH, O, S, CH2, NH(C=O), C(=O)NH or C=O; R 44 , R 45 and R 46 are independently hydrogen, CN or C 1-2 alkyl; t is 0, 1, 2, 3 or 4; Each R e and R f is independently a halogen group, a cyano group, a C 1-4 alkyl group or a C 1-4 haloalkyl group; R 41 is a halogenated group, CN or NO2; R 42 is a halogenated group, CH3, CH2F, CHF2 or CF3; or R 41 and R 42 together form where the dash indicates the bond to ring a; R 43 is hydrogen, a halogenated group, C 1-2 alkyl, C2 alkenyl, NO2, CF3; or R 42 and R 43 together form where each is a single or double bond, and where the dash indicates the bond to ring a.

68. A compound according to any one of claims 1 - 46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 69. A compound according to any one of claims 1 - 46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 70. A compound according to any one of claims 1 - 46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 71. A compound according to any one of claims 1 - 46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is 72. A compound according to any one of claims 1-46, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein B 1 is derived from progesterone, embosamore, bicalutamide, apalutamide, testosterone, dihydrotestosterone, testosterone, 19-nortestosterone, progesterone, ardaline, cortisol, prednisone, flutamide, nilutamide, enzalutamide, tamoxifen, toremifene, raloxifene, bazedoxifene, ospemifene, megestrol acetate, estramustine, abiraterone, LGD-2941, BMS-564929, ostarine, ulipristal acetate, asoprisnil (J867), mifepristone, telapristone (CDB-4124, Proellex, Progenta) or an analogue thereof.

73. A compound according to any one of claims 1-72, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein L 1 has the formula: -(L a ) q -， Wherein: Each L a is independently W, -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -NR 110 S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -C(O)O-, C 1-12 alkylene, C 2-12 alkenylene, C 2-12 alkynylene, C 6-12 arylene, C 3-12 cycloalkylene, 5- to 12-membered hetero-substituted cycloalkylene or 5- to 12-membered heteroarylene, each of which is independently optionally substituted with one or more substituents independently selected from: -OH, -NH2, -CN, oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocyclic group, where each W is independently where R n is independently H, C 1-4 alkyl or C 1-4 haloalkyl; and where R w is independently H, C 3-12 cycloalkyl, C optionally substituted with one or more halo groups or OH 6-12 aryl or C optionally substituted with one or more substituents independently selected from 1-4 alkyl: halo group, OH, -SH, -S(C 1-4 alkyl), -CONH2, -COOH, -NHC(=NH)NH2, -NH2, -NHCOCH3, -NHCHO, -NHCONH2, C 6-12 alkyl, 5-12 membered heterocyclic group or 5-12 membered heteroaryl; Each R 110 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5-12 membered heteroaryl, C 3-12 cycloalkyl or 5-12 membered heterocycloalkyl; Each R 120 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; and q is an integer from 0 to 40.

74. A compound according to any one of claims 1-72, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein L 1 has the formula: -(L a ) q -， Wherein: Each L a is independently -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -NR 110 S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -C(O)O-, C 1-12 alkylene, C 2-12 alkenylene, C 2-12 alkynylene, C 6-12 arylene, C 3-12 cycloalkylene, 5- to 12-membered hetero-substituted cycloalkylene or 5- to 12-membered heteroarylene, each of which is independently optionally substituted with one or more substituents independently selected from: oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocyclic group; Each R 110 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; Each R 120 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5-12 membered heteroaryl, C 3-12 cycloalkyl or 5-12 membered heterocycloalkyl; and q is an integer from 0 to 20.

75. A compound according to any one of claims 1-72, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein L 1 has the formula: -Y 10 -(CHR 130 ) n’ -Y 20 -(CHR 140 ) n” -Y 30 -(CHR 150 ) m” -Y 40 -(CHR 160 ) p -Y 50 -(CHR 170 ) p' -Y 60 -, Wherein: Each Y 10 、Y 20 、Y 30 、Y 40 、Y 50 and Y 60 is independently -(W) s -、a key、-NR 110 -、-O-、-S(O) 0-2 -、-NR 110 C(O)-、-C(O)NR 110 -、-NR 110 C(O)NR 110 -、-NR 110 S(O)2-、-S(O)2NR 110 -、-NR 110 S(O)2NR 110 -、-CR 120 =N-NR 110 -、-NR 110 -N=CR 120 -、-C(O)-、-OC(O)-、-OC(O)O-、-(CH2CH2O) 1-5 -、-C(O)O-、C 1-12 alkylene、C 2-12 alkenylene、C 2-12 alkynylene、C 6-12 arylene、C 3-12 cycloalkylene、a 5- to 12-membered hetero cycloalkylene or a 5- to 12-membered heteroarylene, each of which is independently optionally substituted by one or more substituents independently selected from the following: -OH, -NH2, -CN, an oxo group, a halogen group, C 1-4 alkyl、C 1-4 haloalkyl、C 1-4 alkoxy、C 1-4 haloalkoxy、C 6-12 aryl、a 5- to 12-membered heteroaryl、C 3-12 cycloalkyl and a 5- to 12-membered heterocyclic group; Each W is independently wherein R n is independently H, C 1-4 alkyl or C 1-4 haloalkyl; and wherein R w is independently H, C 3-12 cycloalkyl, C optionally substituted by one or more halo groups or OH 6-12 aryl or C optionally substituted by one or more substituents independently selected from 1-4 alkyl: halo group, OH, -SH, -S(C 1-4 alkyl), -CONH2, -COOH, -NHC(=NH)NH2, -NH2, -NHCOCH3, -NHCHO, -NHCONH2, C 6-12 aryl, 5-12 membered heterocyclic group or 5-12 membered heteroaryl; Each R 110 、R 120 、R 130 、R 140 、R 150 、R 160 and R 170 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5-12 membered heteroaryl, C3-12 cycloalkyl or 5-12 membered heterocyclic group, each of which is independently optionally substituted with one or more substituents independently selected from: -OH, -NH2, -CN, oxo group, halo group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5-12 membered heteroaryl, C 3-12 cycloalkyl and 5-12 membered heterocyclic group; and n', n”, m”, s, p and p' are each independently 0, 1, 2, 3, 4, 5, 6, 7 or 8.

76. A compound according to any one of claims 1-72, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein L 1 has the formula: -Y 10 -(CHR 130 ) n ’-Y 20 -(CHR 140 ) n” -Y 30 -(CHR 150 ) m” -Y 40 - Wherein: Each Y 10 、Y 20 、Y 30 and Y 40 is independently a key, -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -NR 110 S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -(CH2CH2O) 1-5 -, -C(O)O-, C 1-12 alkylene, C 2-12 alkenylene, C 2-12 alkynylene, C 6-12 arylene, C 3-12 cycloalkylene, 5-12 membered hetero-substituted cycloalkylene or 5-12 membered hetero-arylene, each independently optionally substituted with one or more substituents independently selected from: oxo group, halogen group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy or C 1-4 haloalkoxy; Each R 110 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; Each R 120 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; Each R 130 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; Each R 140 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; Each R 150 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl or 5- to 12-membered heterocycloalkyl; and n', n” and m” are each independently 0, 1, 2, 3, 4, 5, 6, 7 or 8.

77. A compound according to any one of claims 1 - 72, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein L 1 has the formula: -L 2 -L 3 -Cy1-L 4 -Cy2-L 5 -L 6 - Wherein: Each L 2 、L 3 、L 4 、L 5 and L 6 is independently a key, C 1-12 alkylene, -O-, -NHC(=O)-, -C(=O)NH-, -C(=O)-O-, -O-C(=O)- or C=O, where one or more carbon atoms in C 1-12 alkylene are optionally replaced by oxygen; Cy1 and Cy2 are each independently a bond, C 6-12 arylene, C 3-12 cycloalkylene, 5- to 12-membered hetero-cycloalkylene or 5- to 12-membered hetero-arylene, where each group is independently optionally substituted with one or more substituents independently selected from: -OH, -NH2, -CN, oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocycloalkyl.

78. The compound according to claim 77, or a stereoisomer thereof, a mixture of stereoisomers, a hydrate, a solvate, an isotope-enriched analogue or a pharmaceutically acceptable salt thereof, wherein Cy1 is a 5-12 membered hetero-subcyclic group optionally substituted by one or more substituents independently selected from: -OH, -NH2, -CN, oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5-12 membered heteroaryl, C 3-12 cycloalkyl and 5-12 membered heterocyclic group.

79. The compound according to claim 77, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein Cy1 is a bond.

80. A compound according to any one of claims 77 - 79, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein Cy2 is a 5 - 12 membered heteroarylene optionally substituted with one or more substituents independently selected from: -OH, -NH2, -CN, oxo, halo, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5 - 12 membered heteroaryl, C 3-12 cycloalkyl and 5 - 12 membered heterocycloalkyl.

81. The compound according to any one of claims 77 - 79, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein Cy2 is a bond.

82. A compound according to any one of claims 1-72, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein L 1 has the formula: Wherein: Each L 2 , L 3 and L 4 is independently a bond, C 1-12 alkylene, -NHC(=O)-, -C(=O)NH-, -C(=O)-O-, -O-C(=O)- or C=O; Each R 200 and R 201 is independently a halogen group, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 6-12 aryl, 5- to 12-membered heteroaryl, C 3-12 cycloalkyl and 5- to 12-membered heterocyclic group; and Each s and s' is independently 0, 1, 2, 3 or 4.

83. The compound according to any one of claims 1 - 72, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein the linking moiety has the following formula: Wherein "*” and the wavy line or dashed line represent covalent bonds.

84. The compound according to any one of claims 1 - 72 or 82, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, wherein the linking moiety has the following formula: Wherein "*” and the wavy line or dashed line represent covalent bonds.

85. A compound, or a stereoisomer, mixture of stereoisomers, hydrate, solvate, isotopically enriched analogue or pharmaceutically acceptable salt thereof, said compound being selected from the compounds in Table 1.

86. A pharmaceutical composition comprising a compound or a stereoisomer thereof, a mixture of stereoisomers, a hydrate, a solvate, an isotope-enriched analogue or a pharmaceutically acceptable salt according to any one of claims 1-85, and a pharmaceutically acceptable excipient.

87. A method for treating cancer, comprising administering to an individual in need thereof an effective amount of a compound according to claims 1-85, or a pharmaceutical composition according to claim 86.

88. The method according to claim 87, wherein the cancer is liver cancer, melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphoblastic leukemia, chronic lymphocytic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, nephroblastoma, cervical cancer, testicular cancer, soft tissue sarcoma, chronic lymphocytic leukemia, Waldenström macroglobulinemia, primary macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, malignant melanoma, small cell lung cancer, gastric cancer, colon cancer, malignant pancreatic insulinoma, malignant carcinoid, malignant melanoma, choriocarcinoma, mycosis fungoides, head and neck cancer, osteogenic sarcoma, pancreatic cancer, acute myeloid leukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma, urogenital cancer, thyroid cancer, esophageal cancer, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, essential thrombocythemia, adrenocortical carcinoma, skin cancer, trophoblastic tumor or prostate cancer.

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