Itentecan-derived topoisomerase-1 inhibitor pharmaceutical composition and application thereof

Abstract

The present invention relates to an alcohol and amine-containing avixetecan derivative, and pharmaceutically acceptable salts or solvates thereof, comprising a structure of formula (I): wherein X is as defined in the disclosure. The disclosure also relates to pharmaceutical compositions comprising these compounds and the use of these compounds and compositions in the prevention or treatment of cancer and / or tumors. # imgabs0 #

Description

Background Art

[0001] The present invention relates to topoisomerase-1 inhibitors derived from the exatecan skeleton, which can be used for tumor treatment. Topoisomerase 1 inhibitors are cytotoxic chemotherapy derivatives with a camptothecin nucleus. Documents reporting the use of the camptothecin derivative, exatecan (chemical name: (1S,9S)-1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3',4':6,7]imidazo[l,2-b]quinoline-10,13(9H,15H)-dione) are disclosed in WO2014057687, US11103593, US9808537, US7091186, US 2010 / 0062008, WO2015057699, Clinical Cancer Research (2016) 22(20):5097-5108, and Cancer Sci (2016) 107:1039-1046. See also WO2022068878, WO2017062271, CN113816969, CN112125915, and US20210353764. Summary of the Invention

[0002] The compounds of the present disclosure represent a class of topoisomerase-1 inhibitors derived from the exatecan skeleton. Specifically, the compounds are described as exatecan amides containing alcohols and amines and their analogs. The compounds are cytotoxic and can be used as chemotherapeutic drugs in tumor treatment, such as as anti-tumor agents. These compounds are powerful and novel in structure and can effectively act on a variety of cancer cell lines.

[0003] For each of the following embodiments, any variable not explicitly defined in the embodiment is defined as in formula (I). In each embodiment described herein, each variable is selected independently of the other variables unless otherwise specified.

[0004] In one embodiment, the present disclosure provides an exatecan derivative containing an alcohol and an amine, and its pharmaceutically acceptable salt, solvate, or stereoisomer, which comprises the structure of formula I:

[0005]

[0006] Wherein:

[0007] R k is selected from hydrogen, -C 1-6 alkyl, (CH2) n C(O)NHC 1-6 alkyl, (CH2) n C 6-10aryl and the alkyl and aryl are optionally substituted with 1 to 3 hydroxyl groups, -C 1-6 alkyl OH groups, and the alkyl is optionally substituted with 1 to 10 halogen groups;

[0008] R j represents hydrogen or C 1-6 alkyl, and the alkyl is optionally substituted with 1 to 10 halogens;

[0009] R 2 and R 3 are independently selected from hydrogen, -C 1-6 alkyl, OH, -C 1-6 alkyl OH, halogen, -C 1-9 haloalkyl, -(CH2) n NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -C 3-6 cycloalkyl, -(CH2) n C 6-10 aryl and -(CH2)nOC 1-6 alkyl, provided that R 2 and R 3 are not simultaneously halogen;

[0010] R 4 is selected from C 1-6 alkyl, OH, -C 1-6 alkyl OH, -CH(OH)C 1-6 alkyl, -C 1-9 haloalkyl, halogen, -C 3-6 cycloalkyl, -(CH2) n OC 1-3 alkyl, -(CH2) n OC 1-9 haloalkyl, -CR x R y C 1-6 alkyl OH, -(CH2) n NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2 and -NHC(O)C 1-6 alkyl NH2;

[0011] R x and R y represent a combination to form a C 3-6 cycloalkyl or spirocycloalkyl C 1-3 alkylene;

[0012] Each n independently represents 0, 1, 2, or 3.

[0013] When R j is hydrogen, one embodiment of the present invention is realized.

[0014] When R j is C 1-6 alkyl, another embodiment of the present disclosure is realized, and the alkyl is optionally substituted with 1 to 5 halogen groups. A sub - embodiment of this aspect of the present invention is realized when R j is C 1-6 alkyl substituted with 1 to 4 halogens.

[0015] When R k is hydrogen, one embodiment of the present disclosure is realized.

[0016] When R k is - C 1-6 alkyl, another embodiment of the present disclosure is realized, and the alkyl is optionally substituted with 1 to 10 halogens or 1 to 3 groups selected from hydroxyl and - C 1-6 alkyl OH.

[0017] When R k is (CH2) n C(O)NHC 1-6 alkyl, another embodiment of the present disclosure is realized, and the alkyl is optionally substituted with 1 to 10 halogens or 1 to 3 groups selected from hydroxyl and - C 1-6 alkyl OH.

[0018] When R k is (CH2) n C 6-10 aryl, another embodiment of the present disclosure is realized, and the aryl is optionally substituted with 1 to 3 groups selected from hydroxyl and - C 1-6 alkyl OH.

[0019] When R k is , another embodiment of the present disclosure is realized, where R 2 、R 3 and R 4 are as described herein.

[0020] One embodiment of the present invention is realized by formula I' or a pharmaceutically acceptable salt, solvate or stereoisomer thereof, and its structural formula is:

[0021]

[0022] When R 2 、R 3 and R 4An embodiment of the present disclosure is achieved when the hydrogen atoms in the alkyl substituents are not deuterated.

[0023] When R 2 , R 3 and R 4 An embodiment of the present disclosure is achieved when at least one hydrogen atom in the alkyl substituents is deuterated.

[0024] Another embodiment of the present disclosure is achieved when n is 0. Another embodiment of the present disclosure is achieved when n is 1. Another embodiment of the present disclosure is achieved when n is 2. Another embodiment of the present disclosure is achieved when n is 3.

[0025] When R x and R y combine with the atoms to which they are attached to form cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, another embodiment of the present disclosure is achieved.

[0026] When R 2 is selected from hydrogen, C 1-6 alkyl, CH2OC 1-6 alkyl, -(CH2) n OH, -CH2F, -CHF2, -CF3, -(CH2)phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -(CH2)nNH2, -NHCH3 and -N(CH3)2, another embodiment of the present disclosure is achieved. When R 2 is selected from hydrogen, C 1-6 alkyl, CH2OCH3, -OH, -CH2OH, -CH2F, -CHF2, -CF3 and -NH2, a sub - embodiment of this aspect of the present disclosure is achieved. When R 2 is hydrogen, another sub - embodiment of this aspect of the present disclosure is achieved. When R 2 is C 1-6 alkyl selected from -CH3, -CH2CH3 and -CH2CH(CH3)2, another sub - embodiment of this aspect of the present disclosure is achieved. When R 2 is (CH2) n OCH3, another sub - embodiment of this aspect of the present disclosure is achieved. When R 2 is -OH or -CH2OH, another sub - embodiment of this aspect of the present disclosure is achieved. When R 2 is -CH2F, -CHF2 or -CF3, another sub - embodiment of this aspect of the present disclosure is achieved. When R 2 is NH2, another sub - embodiment of this aspect of the present disclosure is achieved. When R 2Another sub - embodiment of this aspect of the present disclosure is achieved when any hydrogen atom in the alkyl - substituted group is not deuterated.

[0027] When R 3 is selected from hydrogen, C 1-6 alkyl, CH2OC 1-6 alkyl, -(CH2) n OH, -CH2F, -CHF2, -CF3, -(CH2)phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -(CH2)nNH2, -NHCH3, and -N(CH3)2, another embodiment of the present disclosure is achieved. When R 3 is selected from hydrogen, C 1-6 alkyl, CH2OCH3, -OH, -CH2OH, -CH2F, -CHF2, -CF3, and -NH2, a sub - embodiment of this aspect of the present disclosure is achieved. When R 3 is hydrogen, another sub - embodiment of this aspect of the present disclosure is achieved. When R 3 is C 1-6 alkyl selected from -CH3, -CH2CH3, and -CH2CH(CH3)2, another sub - embodiment of this aspect of the present disclosure is achieved. When R 3 is -CH2OCH3, another sub - embodiment of this aspect of the present disclosure is achieved. When R 3 is -OH or -CH2OH, another sub - embodiment of this aspect of the present disclosure is achieved. When R 3 is -CH2F, -CHF2, or -CF3, another sub - embodiment of this aspect of the present disclosure is achieved. When R 3 is NH2, yet another sub - embodiment of this aspect of the present disclosure is achieved. When R 3 Another sub - embodiment of this aspect of the present disclosure is achieved when any hydrogen atom in the alkyl - substituted group is not deuterated.

[0028] When R 2 and R 3 are both hydrogen, another embodiment of the present disclosure is achieved.

[0029] When at least one of R 2 and R 3 is -OH or -(CH2) n OH (where n is not 0), yet another embodiment of the present disclosure is achieved. When one of R 2 and R 3 is -OH or -(CH2) n OH (where n is not 0), one aspect of the present disclosure is achieved. When R 2 and R 3Another aspect of the present disclosure is achieved when one of them is -OH and the other is not. When R 2 and R 3 one of them is -OH and the other is selected from hydrogen and C 1-6 alkyl, another aspect of the present disclosure is achieved. When R 2 and R 3 one of them is -CH2OH and the other is not, another aspect of the present disclosure is achieved. When R 2 and R 3 one of them is -CH2OH and the other is selected from hydrogen and C 1-6 alkyl, another aspect of the present disclosure is achieved. When R 2 and R 3 one or the other of them contains C 1-6 alkyl and no hydrogen in the alkyl is deuterated, another aspect of the present disclosure is achieved.

[0030] When R 2 and R 3 at least one of them is -(CH2) n NH2, -NHCH3 or -N(CH3)2, another embodiment of the present disclosure is achieved. When R 2 and R 3 at least one of them is (CH2) n NH2, one aspect of this embodiment of the present disclosure is achieved. When R 2 and R 3 one of them is NH2 and the other is not, another aspect is achieved. When R 2 and R 3 one of them is NH2 and the other is selected from hydrogen, CH2 phenyl and C 1-6 alkyl, another aspect of the present disclosure is achieved. When R 2 and R 3 one of them is CH2NH2 and the other is not, another aspect is achieved. When R 2 and R 3 one of them is CH2NH2 and the other is selected from hydrogen, CH2 phenyl and C 1-6 alkyl, another aspect of the present disclosure is achieved. When R 2 and R 3 one or the other of them contains C 1-6 alkyl and no hydrogen in the alkyl is deuterated, another aspect of the present disclosure is achieved.

[0031] When R 4 is selected from C 1-6 alkyl, -C 1-6 alkyl OH, OH, -CH(OH)C 1-6 alkyl, -(CH2)n OC 1-3 alkyl, -C 3-6 cycloalkyl, -C 1-9 haloalkyl, -(CH2) n OC 1-9 haloalkyl, halogen, -CR x R y C 1-6 alkyl OH, -(CH2) n NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2 and -NHC(O)C 1-6 alkyl NH2, another embodiment of the present disclosure is achieved. When R 4 is selected from C 1-6 alkyl, -(CH2) n OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3, fluorine, (CH2)nOCHF2, cyclopropyl, -(CH2) n NH2, -spirocyclopropyl CH2OH and -NHC(O)CH(CH3)NH2, one aspect of this embodiment of the present disclosure is achieved. When R 4 is selected from CH3, -(CH2) n OH or -CH(CH3)OH, another aspect of the present disclosure is achieved. When R 4 is -CH3, another embodiment of this aspect of the present disclosure is achieved. When R 4 is -(CH2) n OH, another embodiment of this aspect of the present disclosure is achieved. When R 4 is -(CH2)2OH or -CH2OH, another embodiment of this aspect of the present disclosure is achieved. When R 4 is -(CH2)2OH, another embodiment of this aspect of the present disclosure is achieved. When R 4 is -CH2OH, another embodiment of this aspect of the present disclosure is achieved. When R 4 is -CH(CH3)OH, another embodiment of this aspect of the present disclosure is achieved. When R 4 is -CH2F, -CHF2 or -CF3, another embodiment of this aspect of the present disclosure is achieved. When R 4 is -(CH2) n NH2, another embodiment of this aspect of the present disclosure is achieved. When R 4 is NH2, another embodiment of this aspect of the present disclosure is achieved. When R 4 is NHC(O)CH(CH3)NH2, another embodiment of this aspect of the present disclosure is achieved. When R 4-CR x R y C 1-6 Another embodiment of this aspect of the disclosure is achieved when R is alkyl OH, where R x and R y are alkylene substituents that combine to form cyclopropyl or spirocyclopropyl. Another embodiment of this aspect of the disclosure is achieved when R 4 is -spirocyclopropyl CH2OH. Another embodiment of this aspect of the disclosure is achieved when R 4 is -(CH2)nOCHF2 or CHF2. Another aspect of the disclosure is achieved when R 4 contains C 1-6 alkyl and no hydrogen in the alkyl is deuterated.

[0032] When one of R 2 and R 3 is -OH or -(CH2) n OH (where n is not 0), and R 4 is selected from C 1-6 alkyl, -C 1-6 alkyl OH, -(CH2) n OC 1-3 alkyl, -CH(OH)C 1-6 alkyl, -C 3-6 cycloalkyl, -C 1-9 haloalkyl, -(CH2) n OC 1-9 haloalkyl, halogen, -CR x R y C 1-6 alkyl OH, -(CH2) n NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, and -NHC(O)C 1-6 alkyl NH2, another embodiment of the disclosure is achieved. When one of R 2 and R 3 is -OH or -CH2OH and the other is selected from hydrogen and C 1-6 alkyl, and R 4 is selected from CH3, -(CH2) n OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3, fluorine, (CH2)nOCHF2, cyclopropyl, -(CH2) n NH2, and -spirocyclopropyl CH2OH, -NHC(O)CH(CH3)NH2, another aspect of the disclosure is achieved. When one of R 2 and R 3 is -OH or -(CH2)n OH (where n is not 0) and the other is selected from hydrogen and C 1-6 alkyl, and R 4 is selected from CH3, -(CH2)2OH, -CH2OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3, fluorine, and -(CH2) n NH2, an embodiment of this aspect of the present disclosure is achieved. When R 2 and R 3 one of them is -OH or -(CH2) n OH (where n is not 0) and the other is selected from hydrogen and C 1-6 alkyl, and R 4 is selected from CH3, -(CH2)2OH, CH2OH, and -(CH2) n NH2, another embodiment of this aspect of the present disclosure is achieved. When n is 0, 1, or 2, an aspect of the present disclosure is achieved. When R 2 , R 3 and / or R 4 contains C 1-6 alkyl and no hydrogen in the alkyl is deuterated, another aspect of the present disclosure is achieved.

[0033] When R 2 and R 3 one of them is -(CH2) n NH2, -NHCH3, or -N(CH3)2 and the other is selected from hydrogen, CH2 phenyl, and C 1-6 alkyl, and R 4 is selected from C 1-6 alkyl, -C 1-6 alkyl OH, -CH(OH)C 1-6 alkyl, -(CH2) n OC 1-3 alkyl, -C 3-6 cycloalkyl, -C 1-9 haloalkyl, -(CH2) n OC 1-9 haloalkyl, halogen, -CR x R y C 1-6 alkyl OH, -(CH2) n NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, and -NHC(O)C 1-6 alkyl NH2, another embodiment of the present invention is achieved. When R 2 and R 3 one of them is (CH2) nNH2, -NHCH3 or -N(CH3)2 and the other is selected from hydrogen, CH2 phenyl and C 1-6 alkyl, and R 4 is selected from CH3, -(CH2) n OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3, fluorine, (CH2)nOCHF2, cyclopropyl, -(CH2) n NH2 and -spirocyclopropyl CH2OH, -NHC(O)CH(CH3)NH2, another aspect of the present disclosure is achieved. When R 2 and R 3 one of them is (CH2) n NH2, -NHCH3 or -N(CH3)2 and the other is selected from hydrogen, CH2 phenyl and C 1-6 alkyl, and R 4 is selected from CH3, -(CH2) n OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3, fluorine and -(CH2) n NH2, an embodiment of this aspect of the present disclosure is achieved. When R 2 and R 3 one of them is (CH2) n NH2, -NHCH3 or -N(CH3)2 and the other is selected from hydrogen, CH2 phenyl and C 1-6 alkyl, and R 4 is selected from (CH2) n OH or -(CH2) n NH2, another embodiment of this aspect of the present disclosure is achieved. When R 4 is -(CH2)2OH, one aspect of the present disclosure is achieved. When R 4 is CH2OH, another aspect of the present disclosure is achieved. When R 4 is (CH2) n NH2, another aspect of the present disclosure is achieved. When R 2 , R 3 and / or R 4 contains C 1-6 alkyl and no hydrogen in the alkyl is deuterated, yet another aspect of the present disclosure is achieved.

[0034] When R 2 and R 3 are both hydrogen and R 4 is selected from CH3, -(CH2) n OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3, fluorine, (CH2)nOCHF2, cyclopropyl, -(CH2) nAnother embodiment of the present disclosure is achieved when R is NH2 and - spirocyclopropyl CH2OH, - NHC(O)CH(CH3)NH2.

[0035] Another embodiment of Formula I and Formula I' of the present disclosure is represented by Structural Formula II:

[0036]

[0037] or a pharmaceutically acceptable salt or solvate thereof, wherein R 4 As described herein. When R 4 is selected from -CH3, -OH, -CH2OH, -(CH2)2OH, -CH(CH3)OH, (CH2)2OCHF2, -C(CH3)2CH2OH, -CH2F, -CHF2, -CF3, fluorine, and -(CH2) n NH2, a sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is -CH3, a sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is -OH, a sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is -CH2OH or -(CH2)2OH, a sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is -CH(CH3)OH, a sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is (CH2)2OCHF2, another sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is -C(CH3)2CH2OH, another sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is -CH2F, -CHF2, or -CF3, another sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is fluorine, a sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is -(CH2) n NH2, a sub - embodiment of Formula II of the present disclosure is achieved. When R 4 contains C 1-6 alkyl and no hydrogen in the alkyl is deuterated, another sub - embodiment of Formula II of the present disclosure is achieved.

[0038] Another embodiment of Formula I and Formula I' of the present disclosure is represented by Structural Formula III:

[0039]

[0040] or a pharmaceutically acceptable salt or solvate thereof, wherein R 4 As described herein. When R 4Selected from CH3, -CH2OH, -(CH2)2OH, -CH(CH3)OH, (CH2)2OCHF2, -C(CH3)2CH2OH, halogen, -CH2F, -CHF2, -OH, -CF3 and -(CH2) n is NH2, a sub - embodiment of Formula III of the present disclosure is achieved. When R 4 is -CH3, a sub - embodiment of Formula III of the present disclosure is achieved. When R 4 is -OH, a sub - embodiment of Formula II of the present disclosure is achieved. When R 4 is -CH2OH or -(CH2)2OH, a sub - embodiment of Formula III of the present disclosure is achieved. When R 4 is -CH(CH3)OH, a sub - embodiment of Formula III of the present disclosure is achieved. When R 4 is (CH2)2OCHF2, another sub - embodiment of Formula III of the present disclosure is achieved. When R 4 is -C(CH3)2CH2OH, another sub - embodiment of Formula III of the present disclosure is achieved. When R 4 is -CH2F, -CHF2 or -CF3, another sub - embodiment of Formula III of the present disclosure is achieved. When R 4 is fluorine, another sub - embodiment of Formula III of the present disclosure is achieved. When R 4 is -(CH2) n NH2, a sub - embodiment of Formula III of the present disclosure is achieved. When R 4 contains C 1-6 alkyl and no hydrogen in the alkyl is deuterated, another sub - embodiment of Formula III of the present disclosure is achieved.

[0041] Another embodiment of Formulas I and I' of the present disclosure is represented by Structural Formula IV:

[0042]

[0043] or a pharmaceutically acceptable salt or solvate thereof, wherein R 4 is as described herein. When R 4 is OH, -CH2F, -CHF2, -CF3, fluorine or (CH2) n NH2, a sub - embodiment of Formula IV of the present disclosure is achieved.

[0044] Another embodiment of Formulas I and I' of the present disclosure is represented by Structural Formula V:

[0045]

[0046] or a pharmaceutically acceptable salt or solvate thereof, wherein R jAs described herein, when R j is C 1-6 alkyl, and the alkyl is optionally substituted with 1 to 10 halogen groups, a sub - embodiment of Formula V of the present disclosure is achieved.

[0047] When R is hydrogen or - C 1-6 alkyl, another embodiment of Formula V is achieved.

[0048] When R k1 is - C 1-6 alkyl, and the alkyl is optionally substituted with 1 to 10 halogens and / or 1 to 3 groups selected from hydroxy and - C 1-6 alkyl OH, another embodiment of Formula V is achieved. When R k1 is - C 1-6 alkyl, and the alkyl is substituted with 1 to 3 groups selected from hydroxy and - C 1-6 alkyl OH and / or 1 to 10 halogens, another embodiment of Formula V is achieved.

[0049] When R k1 is (CH2) n C(O)NHC 1-6 alkyl, and the alkyl is optionally substituted with 1 to 10 halogens and / or 1 to 3 groups selected from hydroxy and - C 1-6 alkyl OH, another embodiment of the present disclosure is achieved.

[0050] When R k1 is (CH2) n C 6-10 aryl, and the aryl is optionally substituted with 1 to 3 groups selected from hydroxy and - C 1-6 alkyl OH, another embodiment of the present invention is achieved.

[0051] One aspect of the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula I, I', II, III, IV, V or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0052] Another aspect of the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula I, I', II, III, IV or V as described herein, or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0053] Another aspect of the present disclosure relates to a compound of formula I, I', II, III, IV or V as described herein, or a tautomer, meso form, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, for use as a medicament or pharmaceutical ingredient.

[0054] Another aspect of the present disclosure relates to a compound of formula I, I', II, III, IV or V as described herein, or a tautomer, meso form, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, for use in the preparation of a medicament for treating or preventing tumors.

[0055] In another embodiment, the compounds of the present disclosure include the compounds described in the examples in the following table and their pharmaceutically acceptable salts. Detailed Description

[0056] The compounds of the present disclosure may contain one or more asymmetric centers and may thus occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. Depending on the nature of the various substituents on the molecule, additional asymmetric centers may exist. Each such asymmetric center will independently give rise to two optical isomers, and it is intended that all possible optical isomers and diastereomers in the mixture, as well as pure or partially purified compounds, are included within the scope of the present disclosure. Unless a specific stereochemistry is indicated, the present disclosure is intended to cover all isomeric forms of these compounds.

[0057] The independent synthesis or chromatographic separation of these diastereomers can be achieved by appropriately modifying the methodologies disclosed herein, according to methods known in the art. Among other methods, their absolute stereochemistry can be determined by X-ray crystallography of the crystalline product or crystalline intermediate, and if desired, the crystalline product or crystalline intermediate can be derivatized with a reagent containing an asymmetric center of known absolute configuration.

[0058] If desired, the racemic mixture of the compound can be separated to isolate the individual enantiomers. The separation can be carried out by methods well known in the art, such as coupling the racemic mixture of the compound with an enantiomerically pure compound to form a diastereomeric mixture, and then separating the individual diastereomers by standard methods (such as fractional crystallization or chromatography). The coupling reaction is usually the formation of a salt using an enantiomerically pure acid or base. The diastereomeric derivative can then be converted to the pure enantiomer by cleavage of the added chiral residue. The racemic mixture of the compound can also be directly separated by chromatography using a chiral stationary phase, which is well known in the art.

[0059] Alternatively, any enantiomer of a compound can be obtained by stereoselective synthesis using optically pure starting materials or reagents of known configuration by methods well known in the art.

[0060] In the compounds of formula I, I', II, III, IV or V, the atoms may exhibit their natural isotopic abundances, or one or more atoms may be artificially enriched with a specific isotope having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. The present disclosure can include all suitable isotopic variants of the compounds of formula I, I', II, III, IV or V. For example, different isotopic forms of hydrogen (H) include protium ( 1 H) and deuterium ( 2 H). Protium is the major hydrogen isotope found in nature. Enrichment with deuterium can provide certain therapeutic advantages, such as increased in vivo half-life or reduced dosage requirements, or can provide compounds that can be used as standards for characterizing biological samples. For the purposes of the present invention, when a compound is referred to as "undeuterated", this means that the deuterium enrichment does not exceed the background state. Isotopically enriched compounds in formulae I, I', II, III, IV or V can be prepared by conventional techniques well known to those skilled in the art or by methods similar to those described in the schemes and examples herein, using appropriate isotopically enriched reagents and / or intermediates, without undue experimentation.

[0061] When the compounds of the present disclosure can form tautomers, all such tautomeric forms are also included within the scope of the present disclosure. For example, a compound containing a carbonyl -CH2C(O)- group (keto form) may tautomerize to form a hydroxy -CH=C(OH)- group (enol form). If present, both the keto and enol forms are included within the scope of the present disclosure.

[0062] When any variable (e.g., R 5 etc.) occurs more than once in any component, its definition at each occurrence is independent of its definition at each other occurrence. In addition, only combinations of substituents and variables that can result in a stable compound are permitted. A line drawn from a substituent into a ring system indicates that the indicated bond can be attached to any substitutable ring atom. If the ring system is bicyclic, it is intended that the bond be attached to any suitable atom on either ring of the bicyclic moiety.

[0063] It should be understood that one or more silicon (Si) atoms can be introduced into the compounds of the present disclosure to replace one or more carbon atoms, thereby providing compounds that are chemically stable and can be readily synthesized from readily available starting materials by techniques known in the art. The covalent radii of carbon and silicon are different, resulting in different bond distances and spatial arrangements when comparing similar C-element and Si-element bonds. These differences result in minor changes in the size and shape of silicon-containing compounds compared to carbon. Those of ordinary skill in the art will understand that the differences in size and shape may result in minor or drastic changes in potency, solubility, lack of off-target activity, packaging properties, etc. (Diass, J.O. et al. Organometallics (2006) 5:1188-1198; Showell, G.A. et al. Bioorganic & Medicinal Chemistry Letters (2006) 16:2555-2558).

[0064] It should be understood that those of ordinary skill in the art can select substituents and substitution patterns on the compounds of the present disclosure to provide compounds that are chemically stable and can be readily synthesized from readily available starting materials by techniques known in the art and the methods described hereinafter. If a substituent itself is substituted by more than one group, it should be understood that these multiple groups can be located on the same carbon or different carbons, as long as a stable structure is produced. The phrase "optionally substituted by one or more substituents" should be understood to mean that the group in question is unsubstituted or can be substituted by one or more substituents.

[0065] Absolute stereochemistry is illustrated by the use of dashed and solid wedge bonds. As shown in Illus-I and Illus-II. Accordingly, the methyl group in Illus-I extends out of the plane of the paper, while the ethyl group in Illus-II extends into the plane of the paper, with the cyclohexene ring in the plane of the paper. It is presumed that the hydrogen atom located on the same carbon atom as the methyl group in Illus-I extends into the plane of the paper, while the hydrogen atom located on the same carbon atom as the ethyl group in Illus-II extends out of the plane of the paper. The convention is the same, where both the dashed and solid rectangles are attached to the same carbon as in Illus-III, the methyl group extends out of the plane of the paper, the ethyl group extends into the plane of the paper, and the cyclohexene ring is in the plane of the paper.

[0066]

[0067] By convention, unless the accompanying text states otherwise, ordinary "stick" bonds or "wavy" bonds represent that all possible stereochemistries are represented, including pure compounds, mixtures of isomers, and racemic mixtures.

[0068] As used herein, unless otherwise specified, the following terms have the following meanings:

[0069] The phrase “at least one,” which is used to refer to the number of components in a composition, such as “at least one pharmaceutical excipient,” means that one member of the designated group is present in the composition and that more than one member may also be present. The components of a composition are generally aliquots of discrete pure materials that are added to the composition, where the purity level of the discrete materials added to the composition is the purity level generally accepted for that type of reagent.

[0070] Whether used to refer to substituents on a compound or components of a pharmaceutical composition, the phrase “one or more” has the same meaning as “at least one”;

[0071] “Effective amount” or “therapeutically effective amount” is intended to describe the amount of at least one compound of the present disclosure or the amount of a composition comprising at least one compound of the present disclosure that effectively treats or inhibits a disease or disorder described herein, thereby producing the desired therapeutic, ameliorating, inhibitory, or prophylactic effect. For example, in the treatment of a central nervous system disease or disorder with one or more of the compounds described herein, an “effective amount” (or “therapeutically effective amount”) means, for example, an amount of at least one compound of Formula I, Formula I’, Formula II, Formula III, Formula IV, or Formula V that results in a therapeutic response in a patient having a central nervous system disease or disorder (“condition”), including a response suitable for controlling, alleviating, ameliorating, or treating the condition or alleviating, ameliorating, reducing, or eliminating one or more symptoms attributable to the condition and / or the long-term stability of the condition, which can be determined, for example, by analyzing pharmacodynamic markers or by clinical evaluation of a patient having the condition;

[0072] “Patient” and “subject” refer to an animal, such as a mammal (e.g., a human), preferably a human;

[0073] “Prodrug” refers to a compound that is rapidly converted in vivo to the parent compound, such as by hydrolysis in the blood, for example, converting a prodrug of Formula I to Formula V to a compound of Formula I, Formula I’, Formula II, Formula III, Formula IV, or Formula V or a salt thereof; detailed discussions are provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series, and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference; the scope of the present disclosure includes prodrugs of the novel compounds of the present disclosure;

[0074] The term "substituted" means that one or more of the recited substituents may occupy one or more of the bonding positions on the substrate that are normally occupied by "-H", provided that such substitution does not exceed the normal valence rules of the atoms in the bonding configuration present in the substrate, and that the substitution ultimately provides a stable compound, that is, the substitution does not provide a compound with substituents that are mutually reactive and located geminally or vicinally to each other; and that the compound provided by the substitution is sufficiently stable to be isolated to a useful purity from the reaction mixture.

[0075] When describing optional substitution in a moiety (e.g., "optionally substituted"), the term means that if a substituent is present, one or more of the recited substituents of the specified substrate may be present at the bonding positions on the substrate that are normally occupied by the default substituent that normally occupies that position. For example, if the default substituent on a carbon atom of an alkyl moiety is a hydrogen atom, the optional substituent may replace the default substituent.

[0076] As used herein, unless otherwise specified, the following terms used to describe moieties, whether including the complete definition of the variable part of the structural representation of the compounds of the present disclosure or substituents attached to the variable part of the structural representation of the group of compounds of the present disclosure, have the following meanings, and unless otherwise specified, the definition of each term (i.e., moiety or substituent) applies to the case where the term is used alone or as a component of another term (e.g., the definition of aryl is the same for aryl and the aryl part of aralkyl, alkaryl, aralkynyl moieties, etc.); moieties are equivalently described herein by structure, printed formula representation, or chemical terms, without meaning any difference in meaning, for example, an "acyl" substituent may be described herein by the term "acyl", the printed formula representation "R'-(C=O)-" or "R'-C(O)-", or the structural representation: Equivalently described, likewise, the use of any or all of these representations does not imply any difference;

[0077] The term "alkyl" (including the alkyl part of other moieties, such as trifluoromethyl-alkyl- and alkoxy-) refers to a straight-chain or branched-chain aliphatic hydrocarbon moiety containing up to about 20 carbon atoms (e.g., the name "C 1-20 -alkyl" denotes an aliphatic hydrocarbon moiety having 1 to 20 carbon atoms). In some embodiments, alkyl preferably contains up to about 10 carbon atoms, unless the term is modified to indicate a shorter chain is contemplated, for example, an alkyl part having 1 to 8 carbon atoms is designated herein as "C 1-8 -alkyl". When the term "alkyl" is represented by two hyphens (i.e., "-alkyl-"), it means that the alkyl part is bonded in such a way that the alkyl part connects the substituents on both sides, for example, "-alkyl-OH" means that the alkyl part connects the hydroxy moiety to the substrate.

[0078] The term "cycloalkyl" refers to a moiety having a principal hydrocarbon chain that forms a monocyclic or bicyclic aliphatic moiety containing at least 3 carbon atoms (the minimum number required to provide a monocyclic moiety), up to a specified maximum number of carbon atoms, typically 8 for a monocyclic moiety and 10 for a bicyclic moiety, including spiro moieties. Examples of cycloalkyl moieties include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. The term "cycloalkyl" also includes non-aromatic fused polycyclic systems containing up to 20 carbon atoms, which may optionally be substituted in accordance with the general definition of "alkyl" herein. Suitable polycyclic cycloalkyls include, for example, but are not limited to: 1-decahydronaphthalene; norbornyl; adamantyl; etc.;

[0079] As used herein, the term "alkylene" refers to a saturated straight-chain or branched-chain aliphatic hydrocarbon group having two residues, said two residues being derived from the removal of two hydrogen atoms from the same carbon atom or two different carbon atoms of the parent alkane. Alkylene is a straight-chain or branched-chain group having 1 to 20 carbon atoms, preferably 1 to 12 carbon atoms, more preferably 1 to 6 carbon atoms. Non-limiting examples are methylene, ethylene, propylene, butylene, pentylene, etc.

[0080] As used herein, when the term "alkyl" is modified by "substituted" or "optionally substituted", it means that one or more C-H bonds in the alkyl moiety are replaced by substituents bonded to the alkyl substrate invoked in defining the moiety or may optionally be replaced.

[0081] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent having 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 8 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc. Polycyclic cycloalkyls include cycloalkyls having spiro, fused, or bridged rings.

[0082] When a bonding line terminating in the middle of the structure is used to represent the bonding between a moiety and a substrate, such as the following representation:

[0083]

[0084] Whether numbered or not, this structure indicates that, unless otherwise defined, the moiety can be bonded to the substrate through any available ring atom (e.g., the numbered atom of the exemplary moiety).

[0085] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or polycyclic fused ring having a conjugated π electron system (i.e., each ring in the system shares a pair of adjacent carbon atoms with another ring in the system), preferably a 6- to 10-membered aryl, such as phenyl, naphthyl, preferably phenyl.

[0086] The term "heteroaryl" refers to an aromatic 5- to 8-membered monocyclic, 8- to 12-membered bicyclic, or 11- to 14-membered tricyclic system, wherein the monocyclic ring has 1 to 3 heteroatoms, the bicyclic ring has 1 to 6 heteroatoms, and the tricyclic ring has 1 to 9 heteroatoms, and the heteroatoms are selected from O, N, or S (e.g., the monocyclic, bicyclic, or tricyclic ring has carbon atoms and 1 to 3, 1 to 6, or 1 to 9 N, O, or S heteroatoms, respectively). Non-limiting examples of heteroaryl are pyridyl, pyrazolyl, pyrimidinyl, furyl, oxazolyl, triazolyl, oxadiazolyl, and phenylthio. The heteroaryl described herein may also contain fused rings sharing common carbon-carbon bonds.

[0087] The term "heterocyclic group" (or heterocycloalkyl) refers to a non-aromatic saturated monocyclic or polycyclic ring system containing 3 to 10 ring atoms, preferably 5 to 10 ring atoms, wherein one or more atoms in the ring system are elements other than carbon, such as nitrogen (e.g., piperidinyl or pyrrolidinyl), oxygen (e.g., furyl and tetrahydropyranyl), or sulfur (e.g., tetrahydrothienyl and tetrahydrothiopyranyl); and wherein the heteroatoms may be alone or in combination, provided that the moiety does not contain adjacent oxygen and / or sulfur atoms present in the ring system; preferably, the heterocyclic group moiety contains 5 to 6 ring atoms; the prefixes aza, oxa, or thia before the heterocyclic group root name indicate that at least one nitrogen, oxygen, or sulfur atom, respectively, is present as a ring atom; the heterocyclic group may optionally be substituted by one or more independently selected substituents;

[0088] The nitrogen or sulfur atom of the heterocyclic group may optionally be oxidized to the corresponding N-oxide, S-oxide, or S,S-dioxide (SO2); non-limiting examples of suitable monocyclic heterocyclic group rings include piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl- (wherein, unless otherwise specified, this moiety is bonded to the substrate through any one of the ring carbon atoms C2, C3, C5, or C6), thiomorpholinyl, thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl, tetrahydrofuryl, tetrahydrothienyl, tetrahydrothiopyranyl, etc.; and polycyclic heterocyclic group compounds, e.g., moieties having the following structures:

[0089] etc.

[0090] The term "solvate" refers to a pharmaceutically acceptable solvate formed by a compound of the present disclosure with one or more solvent molecules. Non-limiting examples of solvent molecules include water, ethanol, acetonitrile, isopropanol, DMSO, ethyl acetate.

[0091] The term "halogen" means fluorine, chlorine, bromine or iodine; unless otherwise specified, preferred halogens are fluorine, chlorine and bromine, and substituents that are halogen atoms mean -F, -Cl, -Br or -I, and "halo-" means a fluorine, chlorine, bromine or iodine substituent bonded to the defined moiety. For example, "haloalkyl" means an alkyl as defined above, wherein one or more bonding positions that are normally occupied by hydrogen atoms on the alkyl moiety are occupied by halogen groups, and perhaloalkyl (or "fully halogenated" alkyl) means that all bonding positions that do not participate in bonding the alkyl substituent to the substrate are occupied by halogen. For example, when the alkyl is selected as methyl, the term perfluoroalkyl means -CF3;

[0092] The terms "hydroxyl" (hydroxyl and hydroxy) mean the HO- group, and "hydroxyalkyl" means a substituent of the formula: "HO-alkyl-", where the alkyl is bonded to the substrate and may be substituted or unsubstituted as defined above; preferred hydroxyalkyl moieties include lower alkyls; non-limiting examples of suitable hydroxyalkyls include hydroxymethyl and 2-hydroxyethyl; and when representing moieties in the text, the bonding order is indicated by a hyphen, for example -alkyl represents a single bond between the substrate and the alkyl moiety, -alkyl-X represents the alkyl bonding the "X" substituent to the substrate, and in structural representations, the bonding order is indicated by a wavy line terminated bond, for example: Indicates that the methylphenyl moiety is bonded to the substrate through the ortho-carbon atom of the methyl substituent, and a bond terminated with a wavy line and drawn into the structure without specifically indicating the atom to which it is bonded indicates that the moiety can be bonded to the substrate via any bondable atom in the moiety, as described in the above example.

[0093] A line - as a bond usually represents a mixture of possible isomers or one of them, for example containing (R)- and (S)-stereochemical configurations.

[0094] In addition, non-wedged bold lines or non-wedged dashed lines are used to depict known relative configurations in structures containing multiple stereocenters. For example:

[0095] and / or

[0096] whereas:

[0097] and / or and / or

[0098] In all cases, compound names are accompanied by the structures drawn and are intended to capture every possible stereochemical arrangement of a given structural isomer based on the synthetic operations employed in its preparation. A list of discrete stereoisomers used or linked indicates that the compound presented (e.g., “Example Number”) is isolated as a single stereoisomer and that the configuration of that stereoisomer corresponds to one of the possible configurations listed. A list of discrete stereoisomers used and linked indicates that the compound presented is isolated as a racemic mixture or a mixture of diastereomers.

[0099] A specific absolute configuration is represented by a bold wedged line or a dashed wedged line. Unless a specific absolute configuration is indicated, this disclosure is intended to cover all such stereoisomeric forms of these compounds.

[0100] In this specification, when there are multiple oxygen and / or sulfur atoms in a ring system, there cannot be adjacent oxygen and / or sulfur atoms in that ring system.

[0101] As is well known in the art, unless otherwise indicated, a bond extending from a particular atom (where no moiety is depicted at the end of the bond) represents a methyl group bonded to the atom through that bond. For example:

[0102] represents

[0103] Unfulfilled valences in the text, schemes, examples, structural formulas, and any tables herein are assumed to have one or more hydrogen atoms in a sufficient number to satisfy the valence.

[0104] One or more compounds of this disclosure may also exist in solvated forms or may optionally be converted to solvates. The preparation of solvates is well known. Thus, for example, M. Caira et al., J. Pharmaceutical Sci., 93(3) , 601 - 611 (2004) describes the preparation of solvates of the antifungal fluconazole in ethyl acetate and from water. Similar preparations of solvates and hemisolvates (including hydrates where the solvent is water or water-based) etc. are described in E.C. van Tonder et al., AAPS PharmSciTech., 5(1),described in article 12(2004) and A.L. Bingham et al., Chem. Commun., 603 - 604(2001). Typical, non - limiting procedures include dissolving the compounds of the present invention in a desired amount of a desired solvent (e.g., an organic solvent, an aqueous solvent, water, or a mixture of two or more thereof) at a temperature above ambient temperature, then cooling the solution (with or without an anti - solvent) at a rate sufficient to form crystals, and then separating the crystals by standard methods. Analytical techniques (e.g., IR spectroscopy) indicate that the solvent (including water) is present in the crystal in the form of a solvate (or a hydrate in the case of water incorporated into the crystal form).

[0105] The present disclosure also includes the compounds of the present disclosure in isolated and purified forms obtained by conventional techniques. The present disclosure is intended to include polymorphs of the compounds of formula I, formula I’, formula II, formula III, formula IV, and formula V, as well as salts, solvates, and prodrugs of the compounds of formula I, formula I’, formula II, formula III, formula IV, and formula V. Certain compounds of the present disclosure may exist in different isomeric forms (e.g., enantiomers, diastereomers, atropisomers). The compounds of the invention include all of their isomeric forms, in pure form and in mixtures of two or more, including racemic mixtures.

[0106] In the same manner, unless otherwise indicated, a structural representation of any tautomeric form of a compound exhibiting tautomerism is intended to include all such tautomeric forms of the compound. Thus, when the compounds of the present disclosure, their salts, and their solvates and prodrugs can exist in different tautomeric forms or are in equilibrium between these forms, all such forms of the compound are covered and included within the scope of the present disclosure. Examples of such tautomers include, but are not limited to, keto / enol tautomeric forms, imine / enamine tautomeric forms, and heteroaromatic forms such as the following moieties:

[0107]

[0108] As used herein, the phrase “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, and that have a reasonable benefit / risk ratio.

[0109] As used herein, the term "pharmaceutically acceptable salt" refers to derivatives of a parent compound modified by preparing acid or base salts thereof. The salts in solid form may exist in more than one crystal structure and may also exist in the form of hydrates. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues (such as amines); basic or organic salts of acidic residues (such as carboxylic acids); and the like. Pharmaceutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include salts derived from inorganic acids such as formic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, etc.; and salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, etc. Salts derived from inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganese salts, manganous salts, potassium salts, sodium salts, zinc salts, etc.

[0110] When the compounds of the present disclosure are basic, salts can be prepared from pharmaceutically acceptable non-toxic acids (including inorganic and organic acids). Such acids include acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc. In one aspect of the present disclosure, the salts are citric acid, hydrobromic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid, fumaric acid, and tartaric acid. Similarly, salts of acidic compounds are formed by reacting with a suitable inorganic or organic base.

[0111] The term "toxic drug" refers to substances that inhibit or prevent cell function and / or cause cell death or destruction. Toxic drugs include toxins and other compounds that can be used for tumor treatment.

[0112] The term "toxin" refers to any substance that may have a harmful effect on cell growth or proliferation. Toxins can be small molecule toxins and their derivatives from bacteria, fungi, plants or animals, including camptothecin derivatives such as irinotecan, maytansinoid and its derivatives (CN101573384) such as DM1, DM3, DM4, auristatin F (AF) and its derivatives such as MMAF, MMAE, 3024 (WO 2016 / 127790 Al, compound 7), diphtheria toxin, exotoxin, ricin A chain, abrin A chain, modeccin, α-patulin, Aleutites fordii toxin, dianthin, Phytolaca americana toxin, Momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin and trichothecenes.

[0113] The term "chemotherapeutic agent" refers to compounds that can be used to treat tumors. This definition also includes antihormonal agents that are used to modulate, reduce, block, or inhibit the hormonal actions that promote cancer growth, usually in the form of systemic or whole-body treatment. They can be hormones. Examples of chemotherapeutic agents include: alkylating agents such as thiotepa; cyclophosphamide (CYTOXAN™); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benaodopa, carboquone, meturedopa, and uredopa; aziridines and methylmelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, trimethylolmelamine; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, nitrobin hydrochloride; melphalan, new nitrogen mustard, estramustine phosphate, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine;Antibiotics, such as aclacinomycin, actinomycin, authramycin, azaserine, bleomycin, cactinomycin C, calicheamicin, carabicin, chromomycin, carzinophilin, chromomycin, actinomycin D, daunorubicin, detorubicin, 6-diazo-5-oxy-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, Marcello-mycin, mitomycin, mycophenolic acid, nogalamycin, olivo-mycin, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin; streptozocin, tuberculocidin, ubenimex, zinostatin, zorubicin; antimetabolites, such as methotrexate, 5-fluorouracil (5-FU); folic acid analogs, such as denopterin, methotrexate, pteropterin, trimetrexate; pterin analogs, such as fludarabine, 6-mercaptopterin, thiomethop-terin, thioguanopterin;Pyrimidine analogs, such as ancitabine, azacitidine, 6-azuridine, carmofur, cytarabine, dideoxyuridine, doxitluridine, enocitabine, floxuridine, 5-FU; androgens, such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenergic drugs, such as aminoglutethimide, mitotane, trilostane; folic acid supplements, such as folinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pintostatin; phenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazide; procarbazine; Razoxane; Sizofiran; Spirogermanium; Tenuazonic acid; Triaziquone; 2,2',2”-Trichloroethylamine; Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitolactol; Pipobroman; Gacytosine; Arabinoside (“Ara-C”); Cyclophosphamide; Thiotepa; Taxanes, such as paclitaxel, Bristol-Myers Squibb Oncology, Princeton, N.J.) and docetaxel, Rhone - Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6 - thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP - 16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT - 11; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoic acid; esperamicins; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing. The definition also includes anti - hormonal agents that modulate or inhibit the effect of hormones on tumors, such as anti - estrogens including tamoxifen, raloxifene, aromatase inhibitor 4(5) - imidazole, 4 - hydroxytamoxifen, trioxifene, keoxifene, LYll 7018, onapristone and Fareston; and anti - androgens such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing.

[0114] As used herein, the term "treatment" or "treating" (e.g., of a disease, disorder or condition or related symptoms, which together or separately may be referred to as an "indication") includes: inhibiting a disease, disorder or condition, i.e., preventing or reducing the disease or its biological processes or its progression or the development of clinical symptoms; or alleviating a disease, i.e., causing regression of the disease or its biological processes or its progression and / or clinical symptoms. "Treatment" as used herein also refers to controlling, ameliorating or reducing the risk of a subject having a disease, disorder or condition associated with a tumor. As used herein, the terms "prevent" or "preventing" a disease, disorder or condition include: preventing the development or progression of the clinical symptoms of the disease, disorder or condition in a mammal that may be exposed to or is susceptible to the disease, disorder or condition but has not yet experienced or shown symptoms of the disease, etc.

[0115] It will be apparent to those skilled in the art that the subjects treated by the methods described herein are typically mammals, including humans and non-human animals (e.g., laboratory animals and companion animals). The term "therapeutically effective amount" refers to the amount of a compound of the present invention that is capable of eliciting a biological or medical response in a tissue, system, animal or human that is sought by a researcher, veterinarian, physician or other clinician.

[0116] As used herein, the term "composition" is intended to encompass a product comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof, and a specified amount of one or more additional specified ingredients, as well as any product directly or indirectly resulting from the combination of the specified amounts of the specified ingredients. This term as related to a pharmaceutical composition is intended to encompass a product comprising an active ingredient (including a compound of the present disclosure or a pharmaceutically acceptable salt thereof), optionally together with one or more additional active ingredients, and an inert ingredient that constitutes a carrier, as well as any product directly or indirectly resulting from the combination, complexation or aggregation of any two or more ingredients, or from the dissociation of one or more ingredients, or from other types of reactions or interactions of one or more ingredients. Thus, the pharmaceutical compositions of the present disclosure encompass any composition made by mixing a compound of the present disclosure or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier. "Pharmaceutically acceptable" means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not harmful to its recipient.

[0117] As described above, other embodiments of the present disclosure each relate to a method of treating a disease, disorder or condition or one or more of its symptoms ("indication"), the method comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising said compound or salt.

[0118] In another embodiment, the present disclosure relates to a method of preparing a medicament for a subject, which comprises combining a compound of the present disclosure or a pharmaceutically acceptable salt thereof with a pharmaceutical carrier or diluent.

[0119] One such embodiment provides a method of treating or preventing cancer selected from the following in a subject in need thereof: breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, gastric cancer, endometrial cancer, salivary gland cancer, esophageal cancer, melanoma, glioma, neuroblastoma, sarcoma, lung cancer (e.g., small cell lung cancer and non-small cell lung cancer), colon cancer, rectal cancer, colorectal cancer, leukemia (e.g., acute lymphoblastic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia), bone cancer, skin cancer, thyroid cancer, pancreatic cancer, and lymphoma (e.g., Hodgkin lymphoma, non-Hodgkin lymphoma, or anaplastic large cell lymphoma), the method comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising the compound, its salt or solvate. In one such embodiment, the subject is human.

[0120] Another aspect of the present disclosure relates to a method of treating and / or preventing tumors, which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising a compound of the present disclosure.

[0121] The method also contemplates combinations with additional therapeutic agents. For example, combinations of the compounds of the present disclosure with PPAR-γ (i.e., PPAR-gamma) agonists and PPAR-δ (i.e., PPAR-delta) agonists can be used to treat certain malignancies. PPAR-γ and PPAR-δ are nuclear peroxisome proliferator-activated receptors gamma and delta. PPAR-γ agonists have been shown to inhibit the angiogenic response to VEGF in vitro; troglitazone and rosiglitazone maleate both inhibit the development of retinal neovascularization in mice (Arch. Ophthamol. 2001; 119:709-717). Examples of PPAR-γ agonists and PPAR-γ / α agonists include, but are not limited to, thiazolidinediones (e.g., DRF2725, CS-011, troglitazone, rosiglitazone, and pioglitazone), fenofibrate, gemfibrozil, clofibrate, GW2570, SB219994, AR-H039242, JTT-501, MCC-555, GW2331, GW409544, NN2344, KRP297, NP0110, DRF4158, NN622, GI262570, PNU182716, DRF552926, 2-[(5,7-dipropyl-3-trifluoromethyl-1,2-benzisoxazol-6-yl)oxy]-2-methylpropanoic acid (disclosed in USSN 09 / 782,856), and 2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-ethylchroman-2-carboxylic acid (disclosed in USSN 60 / 235,708 and 60 / 244,697) or a pharmaceutically acceptable salt thereof.

[0122] Another embodiment of the present disclosure is the use of the compounds of the present disclosure in combination with gene therapy for the treatment of cancer. For an overview of genetic strategies for treating cancer, see Hall et al. (Am. J. Hum. Genet. 61:785-789, 1997) and Kufe et al. (Cancer Medicine, 5th Ed, pp 876-889, BC Decker, Hamilton 2000). Gene therapy can be used to deliver any tumor suppressor gene. Examples of such genes include, but are not limited to, p53, which can be delivered via recombinant virus-mediated gene transfer (e.g., see U.S. Patent No. 6,069,134), uPA / uPAR antagonists ("Adenovirus-Mediated Delivery of a uPA / uPAR Antagonist Suppresses Angiogenesis-Dependent Tumor Growth and Dissemination in Mice," Gene Therapy, August 1998; 5(8):1105-13) and interferon γ (J. Immunol. 2000; 164:217-222).

[0123] The compounds of the present disclosure can also be administered in combination with inhibitors of intrinsic multidrug resistance (MDR), particularly MDR associated with high levels of transporter expression. Such MDR inhibitors include p-glycoprotein (P-gp) inhibitors such as LY335979, XR9576, OC144-093, R101922, VX853 and PSC833 (valspodar), or a pharmaceutically acceptable salt thereof.

[0124] The compounds of the present disclosure can also be administered together with immunostimulating drugs such as levamisole, isoprinosine and Zadaxin, or a pharmaceutically acceptable salt thereof.

[0125] The compounds of the present disclosure can also be combined with P450 inhibitors for treating or preventing cancer, and the P450 inhibitors include: exogenous substances, quinidine, tyramine, ketoconazole, testosterone, quinine, methyrapone, caffeine, phenelzine, doxorubicin, troleandomycin, cyclobenzaprine, erythromycin, cocaine, furafyline, cimetidine, dextromethorphan, ritonavir, indinavir, amprenavir, diltiazem, terfenadine, verapamil, cortisol, itraconazole, mibefradil, nefazodone, and nelfinavir, or pharmaceutically acceptable salts thereof.

[0126] The compounds of the present disclosure can also be combined with Pgp and / or BCRP inhibitors for treating or preventing cancer, and the Pgp and / or BCRP inhibitors include: cyclosporin A, PSC833, GF120918, cremophor EL, fumitremorgin C, Ko132, Ko134, Iressa, Imatnib mesylate, EKI-785, Cl1033, novobiocin, diethylstilbestrol, tamoxifen, resperpine, VX-710, tryprostatin A, flavonoids, ritonavir, saquinavir, nelfinavir, omeprazole, quinidine, verapamil, terfenadine, ketoconazole, nifidepine, FK506, amiodarone, XR9576, indinavir, amprenavir, cortisol, testosterone, LY335979, OC144-093, erythromycin, vincristine, digoxin, and talinolol, or pharmaceutically acceptable salts thereof.

[0127] The compounds of the present disclosure may also be used in combination with bisphosphonates for the treatment or prevention of cancer, including bone cancer. The bisphosphonates include, but are not limited to: etidronate (Didronel), pamidronate (Aredia), alendronate (Fosamax), risedronate (Actonel), zoledronate (Zometa), ibandronate (Boniva), incadronate or cimadronate, clodronate, EB-1053, minodronate, neridronate, piridronate, and tiludronate (including any and all pharmaceutically acceptable salts, derivatives, hydrates, and mixtures thereof).

[0128] The compounds of the present disclosure may also be used in combination with aromatase inhibitors for the treatment or prevention of breast cancer. Examples of aromatase inhibitors include, but are not limited to: anastrozole, letrozole, and exemestane, or pharmaceutically acceptable salts thereof.

[0129] The compounds of the present disclosure may also be used in combination with siRNA therapy for the treatment or prevention of cancer.

[0130] The compounds of the present disclosure can also be administered in combination with γ-secretase inhibitors and / or NOTCH signaling inhibitors. Such inhibitors include the compounds described in the following patents: WO 01 / 90084, WO 02 / 30912, WO 01 / 70677, WO 03 / 013506, WO 02 / 36555, WO 03 / 093252, WO 03 / 093264, WO 03 / 093251, WO 03 / 093253, WO2004 / 039800, WO 2004 / 039370, WO 2005 / 030731, WO 2005 / 014553, USSN 10 / 957,251, WO2004 / 089911, WO 02 / 081435, WO 02 / 081433, WO 03 / 018543, WO 2004 / 031137, WO 2004 / 031139, WO 2004 / 031138, WO 2004 / 101538, WO 2004 / 101539 and WO 02 / 47671 (including LY-450139), or pharmaceutically acceptable salts thereof.

[0131] In one embodiment, the specific anti-cancer agents that can be used in this combination therapy include, but are not limited to: pembrolizumab abarelix (Plenaxis ) ; aldesleukin aldesleukin Alemtuzumab alitretinoin allopurinol altretamine amifostine anastrozole arsenic trioxide asparaginase azacitidine bevacizumab bexarotene capsules bexarotene gel bleomycin bortezomib intravenous busulfan oral busulfan Calusterone Capecitabine Carboplatin Carmustine Carmustine Carmustine and Polifeprosan 20 Implant (Gliadel ); Celecoxib Cetuximab Chlorambucil Cisplatin Cladribine ( 2- ); Clofarabine Cyclophosphamide Cyclophosphamide (Cytoxan ); Cyclophosphamide (Cytoxan ); Cytarabine (Cytosar- ); Liposomal Cytarabine Dacarbazine (DTIC- ); Actinomycin D Darbepoetin alfa Liposomal Daunorubicin Daunorubicin, Adriamycin Daunorubicin, Adriamycin Denileukindiftitox Dexrazoxane Docetaxel Doxorubicin (Adriamycin ); Doxorubicin Doxorubicin (Adriamycin PFS ); Liposomal Doxorubicin Dromostanolone Propionate Dromostanolone Propionate (Masterone ); Elliott's B Solution (Elliott's B );Epirubicin Epoetin alfa Erlotinib Estramustine Etoposide phosphate Etoposide, VP-16 Exemestane Filgrastim Floxuridine (intra-arterial) Fludarabine Fluorouracil, 5-FU Fulvestrant Gefitinib Gemcitabine Gemtuzumab ozogamicin Goserelin acetate (Zoladex );Goserelin acetate Histrelin acetate (Histrelin );Hydroxyurea Ibritumomab Tiuxetan Idarubicin Ifosfamide Imatinib mesylate Interferon α2a (Roferon );Interferon α2b (Intron );Irinotecan Lenalidomide Letrozole Leucovorin Leuprolide Acetate Levamisole Lomustine CCNU Meclorethamine, Nitrogen mustard Megestrol Acetate Melphalan L-PAM Mercaptopurine, 6-MP Mesna Mesnex );Methotrexate Methoxsalen Mitomycin C Mitotane Mitoxantrone Nandrolone Phenpropionate (Durabolin- );Nelarabine Nofetumomab Oprelvekin Oxaliplatin Paclitaxel Paclitaxel Paclitaxel Protein-Bound Particles Palifermin Pamidronate Pegademase (Adagen (Pegademase Bovine) );Pegaspargase Pegfilgrastim Pemetrexed Disodium Pentostatin Pipobroman Plicamycin, Mithramycin Porfimer Sodium Procarbazine Quinacrine Rasburicase Rituximab Ridaforolimus; sargramostim sargramostim sorafenib streptozocin sunitinib maleate talc tamoxifen temozolomide teniposide VM - 26 testolactone thioguanine, 6 - TG thiotepa topotecan toremifene Tositumomab Tositumomab / I - 131 Tositumomab Trastuzumab tretinoin ATRA Uracil Mustard (Uracil Mustard );valrubicin vinblastine vincristine vinorelbine Olaparib vorinostat and zoledronate or a pharmaceutically acceptable salt thereof.

[0132] Accordingly, the scope of the present disclosure encompasses the combined use of the compounds of the present disclosure with a second compound selected from: estrogen receptor modulators, androgen receptor modulators, retinoic acid receptor modulators, cytotoxic / cytostatic agents, anti-proliferative agents, prenyl protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, angiogenesis inhibitors, PPAR-γ agonists, PPAR-δ agonists, inhibitors of intrinsic multidrug resistance, antiemetics, agents useful for treating anemia, agents useful for treating neutropenia, immunopotentiating drugs, inhibitors of cell proliferation and survival signal transduction, bisphosphonates, aromatase inhibitors, siRNA therapeutics, γ-secretase and / or NOTCH inhibitors, agents that interfere with receptor tyrosine kinases (RTKs), agents that interfere with cell cycle checkpoints, and any of the therapeutic agents listed above.

[0133] Another example of the present disclosure is a method of treating cancer, which comprises administering a therapeutically effective amount of a combination of a compound of the present disclosure with paclitaxel or trastuzumab.

[0134] The therapeutic combinations disclosed herein can be used in combination with one or more other active agents, said other active agents including but not limited to other anti-cancer agents for preventing, treating, controlling, ameliorating a particular disease or disorder (e.g., a cell proliferation disorder) or reducing the risk thereof. In one embodiment, the compounds of the present disclosure can be combined with one or more other anti-cancer agents for preventing, treating, controlling, ameliorating or reducing the risk of a particular disease or disorder for which the compounds of the present disclosure are applicable. Such other active agents can be administered before, simultaneously with or sequentially to the compounds of the present invention by the commonly used routes and amounts.

[0135] The present disclosure also includes a pharmaceutical composition useful for treating or preventing cancer, which comprises a therapeutically effective amount of a compound of the present disclosure and a second compound selected from: estrogen receptor modulators, androgen receptor modulators, retinoic acid receptor modulators, cytotoxic / cytostatic agents, anti-proliferative agents, prenyl protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, angiogenesis inhibitors, PPAR-γ agonists, PPAR-δ agonists, inhibitors of cell proliferation and survival signal transduction, bisphosphonates, aromatase inhibitors, siRNA therapeutics, γ-secretase and / or NOTCH inhibitors, agents that interfere with receptor tyrosine kinases (RTKs), agents that interfere with cell cycle checkpoints, and any of the therapeutic agents listed above.

[0136] The present disclosure also relates to a method of treating cancer in a human patient, which comprises administering to the patient a compound of the present disclosure and a PD-1 antagonist. The compound of the present disclosure and the PD-1 antagonist can be administered simultaneously or sequentially.

[0137] In certain embodiments, the PD-1 antagonist is an anti-PD-1 antibody or an antigen-binding fragment thereof. In alternative embodiments, the PD-1 antagonist is an anti-PD-L1 antibody or an antigen-binding fragment thereof. In some embodiments, the PD-1 antagonist is an anti-PD-1 antibody independently selected from the following: pembrolizumab, nivolumab, cemiplimab, sintilimab, tislelizumab, atezolizumab (MPDL3280A), camrelizumab, and toripalimab. In other embodiments, the PD-L1 antagonist is an anti-PD-L1 antibody independently selected from atezolizumab, durvalumab, and avelumab.

[0138] In one embodiment, the PD-1 antagonist is pembrolizumab. In a particular sub-embodiment, the method comprises administering 200 mg of pembrolizumab to the patient approximately every three weeks. In other sub-embodiments, the method comprises administering 400 mg of pembrolizumab to the patient approximately every six weeks.

[0139] In a further sub-embodiment, the method comprises administering 2 mg / kg of pembrolizumab to the patient approximately every three weeks. In a particular sub-embodiment, the patient is a pediatric patient.

[0140] In some embodiments, the PD-1 antagonist is nivolumab. In a specific sub-embodiment, the method comprises administering 240 mg of nivolumab to the patient approximately every two weeks. In other sub-embodiments, the method comprises administering 480 mg of nivolumab to the patient approximately every four weeks.

[0141] In some embodiments, the PD-1 antagonist is cemiplimab. In a specific embodiment, the method comprises administering 350 mg of cemiplimab to the patient approximately every 3 weeks.

[0142] In some embodiments, the PD-1 antagonist is atezolizumab. In a specific sub-embodiment, the method comprises administering 1200 mg of atezolizumab to the patient approximately every three weeks.

[0143] In some embodiments, the PD-1 antagonist is durvalumab. In a specific sub-embodiment, the method comprises administering 10 mg / kg of durvalumab to the patient approximately every two weeks.

[0144] In some embodiments, the PD-1 antagonist is avelumab. In a specific sub - embodiment, the method comprises administering 800 mg of avelumab to a patient approximately every two weeks.

[0145] When the compounds of the present disclosure are administered in combination with an anti - human PD - 1 antibody (or an antigen - binding fragment thereof), the anti - human PD - 1 antibody (or an antigen - binding fragment thereof) can be administered simultaneously with, before, or after the compounds of the present disclosure. The anti - human PD - 1 antibody (or an antigen - binding fragment thereof) and / or the compounds of the present disclosure or their pharmaceutically acceptable salts can be administered separately by the same or different routes of administration, or in the same pharmaceutical composition together with other agents. The weight ratio of the anti - human PD - 1 antibody (or an antigen - binding fragment thereof) to the compounds of the present disclosure can vary and depends on the therapeutically effective dose of each agent. Generally, the therapeutically effective dose of each agent will be used. A combination comprising at least one anti - human PD - 1 antibody (or an antigen - binding fragment thereof), the compounds of the present disclosure, and optionally other active agents will typically comprise a therapeutically effective dose of each active agent. In such combinations, the anti - human PD - 1 antibody (or an antigen - binding fragment thereof), the compound, and the other active agents can be administered separately or in combination. In addition, the administration of one agent can be before, simultaneous with, or after the administration of the other agents.

[0146] In one embodiment, the present disclosure provides an anti - human PD - 1 antibody (or an antigen - binding fragment thereof) and / or the compounds of the present disclosure, and at least one other active agent as a combination formulation for simultaneous, separate, or sequential use in the treatment of cancer.

[0147] The present disclosure also provides the use of the compounds of the present disclosure for the treatment of cancer, wherein the patient has been previously (e.g., within 24 hours) treated with an anti - human PD - 1 antibody (or an antigen - binding fragment thereof). The present disclosure also provides the use of an anti - human PD - 1 antibody (or an antigen - binding fragment thereof) for the treatment of a cell - proliferative disease, wherein the patient has been previously (e.g., within 24 hours) treated with an antibody - linker - payload compound (ADC), i.e., the compounds of the present disclosure.

[0148] The present disclosure also relates to a method for treating cancer, the method comprising administering to a subject in need thereof a combination therapy comprising: (a) the compounds of the present disclosure, and (b) an anti - human PD - 1 antibody (or an antigen - binding fragment thereof); wherein the anti - human PD - 1 antibody (or an antigen - binding fragment thereof) is administered once every 21 days.

[0149] In addition, the present disclosure relates to methods of treating cancer, the methods comprising administering to a subject in need thereof a combination therapy comprising: (a) a compound of the present disclosure, and (b) an anti-human PD-1 antibody (or an antigen-binding fragment thereof). In a specific embodiment, the cancer presents as one or more solid tumors or lymphomas. In a further specific embodiment, the cancer is selected from the group consisting of advanced or metastatic solid tumors and lymphomas. In yet a further specific embodiment, the cancer is selected from the group consisting of: malignant melanoma, head and neck squamous cell carcinoma, MSI-H cancer, MMR-deficient cancer, non-small cell lung cancer, urothelial cancer, gastric or gastroesophageal junction adenocarcinoma, breast cancer, and lymphoma. In another embodiment, the group consisting of: diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, small lymphocytic lymphoma, mediastinal large B-cell lymphoma, splenic marginal zone B-cell lymphoma, extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (malt), nodal marginal zone B-cell lymphoma, lymphoplasmacytic lymphoma, primary effusion lymphoma, Burkitt lymphoma, anaplastic large cell lymphoma (primary cutaneous type), anaplastic large cell lymphoma (systemic), peripheral T-cell lymphoma, angioimmunoblastic T-cell lymphoma, adult T-cell lymphoma / leukemia, nasal-type extranodal NK / T-cell lymphoma, enteropathy-associated T-cell lymphoma, γ / δ hepatosplenic T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, mycosis fungoides, and Hodgkin lymphoma. In a particular embodiment, the cell proliferative disease is a metastasized cancer, such as liver metastasis of colorectal cancer. In another embodiment, the cell proliferative disease is a cancer classified as stage III cancer or stage IV cancer. In the case of these embodiments, the cancer cannot be resected surgically.

[0150] In an embodiment of the methods disclosed herein, the anti-human PD-1 antibody (or an antigen-binding fragment thereof) is administered by intravenous infusion or subcutaneous injection.

[0151] In one embodiment, the present disclosure provides a composition comprising a compound of the present disclosure, a pharmaceutically acceptable carrier, and an anti-human PD-1 antibody (or an antigen-binding fragment thereof).

[0152] In another embodiment, the present disclosure provides a composition comprising a compound of the present disclosure, a pharmaceutically acceptable carrier, and pembrolizumab.

[0153] In one embodiment, the present disclosure provides a composition comprising a compound of the present disclosure, a pharmaceutically acceptable carrier, and two additional therapeutic agents, wherein one therapeutic agent is an anti-human PD-1 antibody (or an antigen-binding fragment thereof) and the other therapeutic agent is independently selected from the group consisting of anticancer agents.

[0154] The compounds of the present disclosure can be used in combination with antiemetics for the treatment of nausea or vomiting, including acute, delayed, late, and anticipatory vomiting, which may be caused by the use of the compounds of the present disclosure alone or in combination with radiotherapy. To prevent or treat vomiting, the compounds of the present disclosure can be used in combination with other antiemetics, especially neurokinin-1 receptor antagonists, 5HT3 receptor antagonists (such as ondansetron, granisetron, tropisetron, and zatisetron), GABAB receptor agonists (such as baclofen), corticosteroids (such as Decadron (dexamethasone), Kenalog, Aristocort, Nasalide, Preferid, Benecorten, or others, such as those disclosed in U.S. Patent Nos. 2,789,118, 2,990,401, 3,048,581, 3,126,375, 3,929,768, 3,996,359, 3,928,326, and 3,749,712), antidopaminergic drugs (such as phenothiazines (such as prochlorperazine, fluphenazine, thioridazine, and mesoridazine), metoclopramide, aprepitant, fosaprepitant, or dronabinol). In another embodiment, the use of antiemetics selected from neurokinin-1 receptor antagonists, 5HT3 receptor antagonists, and corticosteroids for combination therapy to treat or prevent vomiting that may be caused after administration of the compounds of the present disclosure is disclosed.

[0155] The compounds of the present disclosure can also be administered together with agents useful for the treatment of anemia. Such anemia-treating agents are, for example, continuous erythropoietin receptor activators (such as epoetin alfa).

[0156] The compounds of the present disclosure can also be administered together with agents for the treatment of neutropenia. Such neutropenia-treating agents are, for example, hematopoietic growth factors that regulate the production and function of neutrophils, such as human granulocyte colony-stimulating factor (G-CSF). Examples of G-CSF include filgrastim.

[0157] The compounds of the present disclosure may be useful when co-administered with other treatment modalities, including but not limited to radiotherapy, surgery, and gene therapy. Thus, in one embodiment, unless otherwise stated, the methods for treating cancer described herein may optionally include administering an effective amount of radiotherapy. For radiotherapy, gamma radiation is preferably used.

[0158] The methods for treating cancer described herein may optionally include administering an effective amount of radiation (i.e., the methods for treating cancer described herein optionally include administering radiotherapy).

[0159] The methods for treating cancer described herein include methods for treating cancer that include administering a therapeutically effective amount of a compound of formula IV in combination with radiotherapy and / or in combination with a second compound selected from: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic / cytostatic agents, anti-proliferative agents, prenyl protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, angiogenesis inhibitors, PPAR-γ agonists, PPAR-δ agonists, inhibitors of intrinsic multidrug resistance, antiemetics, agents useful for treating anemia, agents useful for treating neutropenia, immunostimulatory drugs, inhibitors of cell proliferation and survival signal transduction, bisphosphonates, aromatase inhibitors, siRNA therapeutics, γ-secretase and / or NOTCH inhibitors, agents that interfere with receptor tyrosine kinases (RTKs), agents that interfere with cell cycle checkpoints, and any additional therapeutic agents listed above.

[0160] Additional embodiments of the present disclosure include the pharmaceutical compositions, combinations, uses, and methods listed above, where it should be understood that each embodiment may be combined with one or more other embodiments to the extent that such combination is consistent with the description of the embodiments. It should also be understood that the embodiments provided above should be understood to include all embodiments, including embodiments obtained by combining the embodiments.

[0161] Kit

[0162] In one aspect, a kit is provided that contains a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt, solvate, or ester of the compound and a pharmaceutically acceptable carrier, vehicle, or diluent.

[0163] In another aspect, a kit is provided that includes an amount of a compound of the present disclosure and an amount of at least one additional therapeutic agent listed above, wherein the amounts of two or more active ingredients produce a desired therapeutic effect. In one embodiment, the compound of the present disclosure and one or more additional therapeutic agents are provided in the same container. In one embodiment, the compound of the present disclosure and one or more additional therapeutic agents are provided in separate containers.

[0164] The present disclosure includes prodrugs of the compounds of the present disclosure within its scope. Generally, such prodrugs will be functional derivatives of the compounds of the present disclosure that are readily convertible in vivo to the desired compound. Accordingly, in the methods of treatment of the present disclosure, the term "administering" or "giving" a compound shall cover the use of the specifically disclosed compound or a compound that may not be specifically disclosed but that is convertible in vivo to the designated compound upon administration to a patient. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs," ed. H. Bundgaard, Elsevier, 1985. Metabolites of these compounds include the active substances that are produced after the compounds of the present disclosure are introduced into a biological environment.

[0165] The compounds described herein or their pharmaceutically acceptable salts and / or solvates can be administered alone, in combination with other compounds of the present disclosure, and / or in combination with other therapeutic agents. The choice of therapeutic agent that can be co-administered with the compounds of the present disclosure will depend in part on the disorder being treated.

[0166] The compounds of the present disclosure can be administered by oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV, intracisternal injection or infusion, subcutaneous injection or implantation), by inhalation spray, nasal, vaginal, rectal, sublingual, buccal, or topical routes of administration, and can be formulated, alone or together, into suitable dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants, and vehicles appropriate for each route of administration. In addition to treating warm-blooded animals, the compounds of the present disclosure are also effective for humans.

[0167] A pharmaceutical composition for administering the compounds of the present disclosure can be conveniently presented in dosage unit form and can be prepared by any method well known in the pharmaceutical art. All methods include the step of combining the active ingredient with a carrier composed of one or more accessory ingredients. Generally, the pharmaceutical composition is prepared by uniformly and intimately mixing the active ingredient with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product into the desired formulation. The amount of the active compound contained in the pharmaceutical composition is sufficient to produce the desired effect on the course or condition of the disease. As used herein, the term "composition" is intended to cover a product containing the specified amounts of the specific ingredients, as well as any product directly or indirectly resulting from the combination of the specified amounts of the specific ingredients.

[0168] The pharmaceutical composition containing the active ingredient can be in a form suitable for oral use, such as tablets, lozenges, troches, aqueous or oily suspensions, dispersible powders or granules, emulsions, solutions, hard or soft gelatin capsules, or syrups or elixirs. Compositions for oral use can be prepared by any method known in the art for preparing pharmaceutical compositions, and such compositions can contain one or more agents selected from sweetening agents, flavoring agents, coloring agents, and preservatives to provide a pharmaceutically elegant and palatable preparation. Tablets contain a mixture of the active ingredient and non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets. These excipients can be, for example: inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binding agents such as starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. Tablets can be uncoated or can be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period. For example, delayed-release materials such as glyceryl monostearate or glyceryl distearate can be employed. They can also be coated by the techniques described in U.S. Patents 4,256,108, 4,166,452, and 4,265,874 to form osmotic therapeutic tablets for controlled release. Oral tablets can also be formulated as rapid-release agents, such as fast-dissolving tablets or wafers, quick-dissolving tablets, or fast-dissolving films.

[0169] The preparations for oral use can also be in the form of hard gelatin capsules, in which the active ingredient is mixed with an inert solid diluent (such as calcium carbonate, calcium phosphate, or kaolin), or in the form of soft gelatin capsules, in which the active ingredient is mixed with water or an oily medium (such as peanut oil, liquid paraffin, or olive oil).

[0170] The aqueous suspension contains a mixture of an active substance and an excipient suitable for the manufacture of an aqueous suspension. Such excipients are suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum and gum arabic; the dispersing agent or wetting agent can be a naturally occurring phospholipid, such as lecithin, or a condensation product of an alkylene oxide and a fatty acid, such as polyoxyethylene stearate, or a condensation product of ethylene oxide and a long-chain fatty alcohol, such as heptadecaethyleneoxycetanol, or a condensation product of ethylene oxide and a partial ester derived from a fatty acid and a hexitol, such as polyoxyethylene sorbitan monooleate, or a condensation product of ethylene oxide and a partial ester derived from a fatty acid and a hexitol anhydride, such as polyvinyl sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more colorants, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

[0171] The oily suspension can be formulated by suspending the active ingredient in a vegetable oil, such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil, such as liquid paraffin. The oily suspension may contain a thickening agent, such as beeswax, hard paraffin or acetyl alcohol. Sweetening agents and flavoring agents, such as those described above, can be added to provide a palatable oral preparation. These compositions can be preserved by adding an antioxidant, such as ascorbic acid.

[0172] Dispersible powders and granules suitable for the preparation of an aqueous suspension by the addition of water provide a mixture of an active ingredient and a dispersing agent or wetting agent, a suspending agent and one or more preservatives. Suitable dispersing agents or wetting agents and suspending agents are, for example, those already mentioned above. Additional excipients, such as sweetening agents, flavoring agents and colorants, may also be present.

[0173] The pharmaceutical compositions of the present disclosure may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin or a mixture thereof. Suitable emulsifying agents can be naturally occurring gums, such as gum arabic or tragacanth gum, naturally occurring phospholipids, such as soy lecithin, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of said partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweetening agents and flavoring agents.

[0174] Syrups and elixirs can be formulated with sweetening agents, such as glycerol, propylene glycol, sorbitol or sucrose. Such preparations may also contain emollients, preservatives, flavoring agents and colorants.

[0175] The pharmaceutical composition can be in the form of a sterile injectable aqueous or oily suspension. The suspension can be formulated according to known techniques using the appropriate dispersing or wetting agents and suspending agents described above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3 - butanediol. Acceptable carriers and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile non - volatile oils are commonly used as solvents or suspending media. For this purpose, any mild non - volatile oil can be employed, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can also be used in the preparation of injectables.

[0176] The compounds of the present disclosure can also be administered in the form of suppositories for rectal drug administration. These compositions can be prepared by mixing the drug with a suitable non - irritant excipient that is solid at room temperature but liquid at rectal temperature, and thus will melt in the rectum and release the drug. Such materials are cocoa butter and polyethylene glycol.

[0177] For topical use, creams, ointments, gels, solutions, or suspensions containing the compounds of the present disclosure can be used. Similarly, transdermal patches can also be used for topical administration.

[0178] The pharmaceutical compositions and methods of the present disclosure can also contain other therapeutically active compounds commonly used for treating the above - mentioned pathological conditions as described herein.

[0179] In the treatment, prevention, control, improvement, or reduction of the risk of the conditions disclosed herein, the appropriate dosage level of the compounds of the present disclosure is generally about 0.01 to 500 mg per kilogram of patient body weight per day, which can be administered in single or multiple doses. Suitable dosage levels can be about 0.01 to 250 mg / kg per day, about 0.05 to 100 mg / kg per day, or about 0.1 to 50 mg / kg per day. Within this range, the dosage can be 0.05 to 0.5, 0.5 to 5, or 5 to 50 mg / kg per day. For oral administration, the composition can be provided in tablet form, containing 1.0 to 1000 mg of the active ingredient, particularly 1.0, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 mg of the active ingredient for symptomatic adjustment of the dosage administered to the patient to be treated. The compound can be administered according to a regimen of 1 to 4 times per day, or can be administered once or twice a day.

[0180] However, it should be understood that for any particular patient, the specific dosage levels and frequency of administration may vary and depend on multiple factors, including the activity of the specific compound employed, the metabolic stability and duration of action of the compound, age, body weight, general health status, sex, diet, mode and time of administration, excretion rate, drug combination, the severity of the particular disorder, and the host being treated.

[0181] The following schemes and examples illustrate methods for preparing the compounds of the present disclosure. The starting materials are prepared according to procedures known in the art or as illustrated herein.

[0182] Preparation Example

[0183] The compounds of the present disclosure can be prepared according to the following schemes and specific examples or modifications thereof, using readily available starting materials, reagents, and conventional synthetic procedures. Variations known to those of ordinary skill in the art but not detailed herein can also be utilized. The general methods for preparing the compounds claimed in the present disclosure can be readily understood by those skilled in the art by reviewing the following schemes and descriptions. Abbreviations used in the experiments may include, but are not limited to, the following:

[0184]

[0185]

[0186] General experimental information:

[0187] Unless otherwise stated, all reactions were carried out with magnetic stirring. Unless otherwise stated, all reagents and solvents were purchased from commercial sources and used as received. When applicable, MeCN / water gradients with TFA, formic acid, or NH4HCO3 modifiers were used, and the reaction progress and analysis of synthetic intermediates were evaluated by LCMS (UV detection with ESI, APCI, or other mass spectrometric detectors). Silica gel and reverse-phase flash column chromatography were performed using commercially available pre-packed columns. Reverse-phase preparative HPLC purification was carried out on a preparative HPLC instrument equipped with UV and MS detection, using MeCN / water gradients with TFA, formic acid, or NH4OH modifiers. Collections were made at room temperature 1 1H NMR spectra, and chemical shifts were reported in ppm relative to the residual protium-solvent signal, and multiplicities, coupling constants (if applicable), and signal integrations were listed. Unless otherwise stated, all EC 50 data shown in the tables refer to the CellTiter- 2.0 cytotoxicity assay described in the biological assay section.

[0188] Synthetic schemes, intermediates, and examples

[0189] The compounds of the present disclosure can be prepared by methods known in the art of organic synthesis, and are given in part by the following general synthetic schemes and specific preparation examples. The starting materials are commercially available or can be prepared by known methods.

[0190] Example 1

[0191] Preparation of Compound 1

[0192]

[0193] To a solution of (R)-3-hydroxydihydrofuran-2(3H)-one (1b, 200 mg, 2.0 mmol) in MeOH (0.20 mL) and DMF (0.40 mL) was added a mixture of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ammonium methanesulfonate (1a, 0.40 mL, 0.12 - 0.16 mmol, ~0.35 - 0.40 M in DMF containing Hunig’s base). The reaction mixture was heated to 70 °C for 5 nights. The mixture was cooled to room temperature, filtered through a syringe filter, and then subjected to reverse-phase column chromatography (25 - 60% MeCN / water, containing 0.1% formic acid modifier) to afford (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,4-dihydroxybutyramide (1) as a solid (Peak 1 was the desired product, and minor isomer product Peak 2 was also detected). MS: m / z = 538 [M+H]. 1 1H NMR (500 MHz, DMSO-d6): δ 8.44 (d, J = 9.0 Hz, 1H), 7.76 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.59–5.54 (m, 1H), 5.41 (s, 2H), 5.24–5.07 (m, 2H), 4.11 (dd, J = 9.1, 3.4 Hz, 1H), 3.59–3.54 (m, 2H), 3.25–3.08 (m, 3H), 2.38 (s, 3H), 2.23–2.10 (m, 2H), 2.03–1.94 (m, 1H), 1.91–1.74 (m, 3H), 0.87 (t, J = 7.3 Hz, 3H).

[0194] The following compounds of the present disclosure in Table 1 were prepared using a method similar to that described in Example 1 with different reaction times (2 - 3 days, and 60 - 65 °C) and replacing the appropriate reactants and / or reagents:

[0195] Table 1

[0196]

[0197] Intermediate I-1c

[0198] Preparation of Intermediate I-1c

[0199]

[0200] To a solution of (R)-3-hydroxy-dihydrofuran-2(3H)-one (1b, 2.2 g, 22 mmol) in MeOH (20 mL) was added Dowex 50W X8 (hydrogen form, strongly acidic), 200 - 400 mesh resin (1.1 g). The reactants were stirred at room temperature for 2 hours and 20 minutes. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting (R)-methyl 2,4-dihydroxybutyrate (I-1a) was used directly as an oil in the next step. 1H NMR (500 MHz, CDCl3): δ 4.40 (dd, J = 7.8, 3.9 Hz, 1H), 3.91–3.81 (m, 2H), 3.81 (s, 3H), 2.13–2.04 (m, 1H), 1.96–1.88 (m, 1H). 1 1H NMR (500 MHz, CDCl3): δ 4.40 (dd, J = 7.8, 3.9 Hz, 1H), 3.91–3.81 (m, 2H), 3.81 (s, 3H), 2.13–2.04 (m, 1H), 1.96–1.88 (m, 1H).

[0201] Step B - Synthesis of Intermediate I-1b

[0202] To a mixture of (R)-methyl 2,4-dihydroxybutyrate (I-1a, 2.0 g, 15 mmol) and potassium acetate (6.0 g, 61 mmol) in DCM (10 mL) and water (10 mL) was added (bromodifluoromethyl)trimethylsilane (4.0 mL, 26 mmol). The reaction was stirred vigorously at room temperature for three nights. The mixture was diluted with saturated NaHCO3, DCM, and 3:1 CHCl3:IPA (100 ml). The resulting mixture was shaken and then passed through a hydrophobic membrane phase separator. The remaining aqueous layer was re-extracted with 3:1 CHCl3:IPA (100 ml), and the mixture was passed through a hydrophobic membrane phase separator. The combined organic layers were concentrated under reduced pressure. The crude material was subjected to silica gel column chromatography (0 - 100% EtOAc / hexane) to give (R)-methyl 4-(difluoromethoxy)-2-hydroxybutyrate (I-1b) as an oil. 1HNMR(500MHz,CDCl3)δ6.18(t,J=74.9Hz,1H),4.35–4.29(m,1H),4.06–3.98(m,2H),3.81(s,3H),2.85(d,J=4.9Hz,1H),2.20–2.13(m,1H),2.02–1.93(m,1H).

[0203] Step C - Synthesis of Intermediate I-1c

[0204] To a solution of methyl (R)-4-(difluoromethoxy)-2-hydroxybutyrate (I-1b, 19 mg, 0.10 mmol) in DCE (2.0 mL) was added trimethyltin hydroxide (75 mg, 0.41 mmol). The reaction mixture was stirred at 60 °C overnight. The mixture was cooled, diluted with EtOAc and 1 M HCl and shaken. The organic layer was separated and passed through a hydrophobic membrane phase separator. Then the organic layer was concentrated under reduced pressure. The resulting (R)-4-(difluoromethoxy)-2-hydroxybutyric acid (I-1c) was used directly as a crude product in the next reaction described in Example 3.

[0205] Intermediate I-2d

[0206] Preparation of Intermediate I-2d

[0207]

[0208] Step A - Synthesis of Intermediate I-2b

[0209] To a mixture of 3-(tert-butoxy)-2-cyclopropyl-3-oxopropanoic acid (I-2a, 0.10 g, 0.50 mmol) and CDI (0.12 g, 0.75 mmol) was added DCM (2.0 mL). The reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with DCM and water, extracted, and then passed through a hydrophobic membrane phase separator. The organic layer was concentrated under reduced pressure to give crude tert-butyl 2-cyclopropyl-3-(1H-imidazol-1-yl)-3-oxopropanoate (I-2b), which was used directly in the next reaction. 1 H NMR(500MHz,CDCl3)δ8.18(s,1H),7.49(s,1H),7.11(s,1H),3.08(d,J=9.7Hz,1H),1.60–1.52(m,1H),1.42(s,9H),0.82–0.72(m,2H),0.53–0.46(m,1H),0.30–0.24(m,1H).

[0210] Step B - Synthesis of Intermediate I-2c

[0211] To a solution of NaBH4 (0.060 g, 1.5 mmol) in THF (2.0 mL) and water (1.5 mL) was added dropwise a solution of tert-butyl 2-cyclopropyl-3-(1H-imidazol-1-yl)-3-oxopropionate (I-2b, 0.13 g, 0.5 mmol) in THF (2.0 mL). The reaction was stirred overnight at room temperature. Then the reaction was diluted with DCM, water and 1 M HCl (3.0 mL). The mixture was extracted and passed through a hydrophobic membrane phase separator. The organic layer was concentrated under reduced pressure to give crude tert-butyl 2-cyclopropyl-3-hydroxypropionate (I-2c), which was used directly in the next step. 1 H NMR (500 MHz, CDCl3) δ 3.86–3.78 (m, 2H), 1.74–1.68 (m, 1H), 1.48 (s, 9H), 0.95–0.86 (m, 1H), 0.57–0.52 (m, 2H), 0.40–0.35 (m, 1H), 0.20–0.16 (m, 1H).

[0212] Step C - Synthesis of Intermediate I-2d

[0213] To a solution of tert-butyl 2-cyclopropyl-3-hydroxypropionate (I-2c, 0.090 g, 0.50 mmol) in DCM (4.0 mL) was added HCl (0.40 mL, 1.6 mmol, 4.0 M in dioxane). The reaction was stirred overnight at room temperature. The mixture was concentrated under reduced pressure, and the resulting crude 2-cyclopropyl-3-hydroxypropanoic acid (I-2d) was used directly in the next step described in Example 3.

[0214] Intermediates I-3d and I-3e

[0215] Preparation of Intermediates I-3d and I-3e

[0216]

[0217] SFC separation of enantiomers gave I-3b and I-3c

[0218] Racemic 2-(((benzyloxy)carbonyl)amino)-3,3-difluoro-2-methylpropanoic acid (I-3a, 1.2 g, 4.4 mmol) was subjected to preparative chiral SFC (AD-H 2x25 cm, 10% (EtOH + 0.1% DEA) / CO2, 100 bar, 70 mL / min) to give enantiomer 1 (peak 1, 95% ee, absolute configuration unknown) and enantiomer 2 (peak 2, 99% ee, absolute configuration unknown) as the diethylamine salts.

[0219] To obtain the free acid, the diethylamine salt of enantiomer 1 was dissolved in EtOAc and shaken with 1 M aqueous HCl. The organic layer was back-extracted twice with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give 2-(((benzyloxy)carbonyl)amino)-3,3-difluoro-2-methylpropanoic acid (I-3b, enantiomer 1, absolute configuration unknown). 1 H NMR (500 MHz, DMSO-d6) δ 13.20 (s, 1H), 8.10 (s, 1H), 7.42–7.27 (m, 5H), 6.28 (t, J = 56.1 Hz, 1H), 5.03 (s, 2H), 1.34 (s, 3H).

[0220] The free acid of 2-(((benzyloxy)carbonyl)amino)-3,3-difluoro-2-methylpropanoic acid (I-3c, enantiomer 2, absolute configuration unknown) was obtained in a similar manner.

[0221] Step A - Synthesis of intermediates I-3d and I-3e

[0222] A mixture of 2-(((benzyloxy)carbonyl)amino)-3,3-difluoro-2-methylpropanoic acid (I-3b, enantiomer 1, 290 mg, 1.1 mmol), 10 wt% Pd / C (430 mg, 0.41 mmol Pd), and MeOH (20 mL) was stirred overnight under H2 (1 atm). The reaction mixture was then filtered, rinsed with MeOH and DCM, and the solvent was removed under reduced pressure to give 2-amino-3,3-difluoro-2-methylpropanoic acid (I-3d, enantiomer 1, absolute configuration unknown) as a solid. H NMR (500 MHz, DMSO-d6) δ 7.71 (br s, 3H), 6.18 (t, J = 54.4 Hz, 1H), 1.27 (s, 3H). 1

[0223] 2-Amino-3,3-difluoro-2-methylpropanoic acid (I-3e, enantiomer 2, absolute configuration unknown) was obtained from I-3c in a similar manner.

[0224] Intermediate I-4b

[0225]

[0226]

[0227] ​​A mixture of ((9H-fluoren-9-yl)methoxy)carbonyl)-L-alanine (I-4a, 31 mg, 0.10 mmol), HATU (36 mg, 0.095 mmol) and Hunig's base (18 μL, 0.10 mmol) in DMF (0.50 mL) was stirred at room temperature for 2 minutes. At this time, 2-amino-3,3-difluoro-2-methylpropanoic acid (I-3d, 14 mg, 0.10 mmol) and additional Hunig's base (18 μL, 0.10 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour and then subjected to reverse-phase column chromatography (20 - 60% MeCN / H2O, containing 0.1% formic acid modifier) to give 2-((S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-3,3-difluoro-2-methylpropanoic acid (I-4b) as a solid. MS: m / z = 455 [M+Na]. 1 1H NMR (500 MHz, DMSO-d6) δ 13.02 (s, 1H), 8.47 (s, 1H), 7.89 (d, J = 7.5 Hz, 2H), 7.73 (t, J = 7.6 Hz, 2H), 7.53 (s, 1H), 7.42 (t, J = 7.4 Hz, 2H), 7.33 (t, J = 7.4 Hz, 2H), 6.29 (t, J = 58.5 Hz, 1H), 4.31–4.17 (m, 3H), 4.15–4.04 (m, 1H), 1.37 (s, 3H), 1.21 (d, J = 7.1 Hz, 3H).

[0228] Intermediate I-5b

[0229] Preparation of Intermediate I-5b

[0230]

[0231] Step A - Synthesis of Compound I-5b

[0232] At -78 °C, a stirred solution of 1,1-difluoropropan-2-ol (I-5a, 0.21 mL, 2.5 mmol) in DCM (1.2 mL) was treated successively with triethylamine (1.1 mL, 7.5 mmol) and nonafluorobutanesulfonyl fluoride (0.76 mL, 4.3 mmol). The reaction mixture was gradually warmed to room temperature and stirred at room temperature overnight. Then the mixture was poured into aqueous Na2CO3 and extracted twice with DCM. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure to give 1,1-difluoropropan-2-yl 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulfonate (I-5b), which was used directly as a crude product.

[0233] Example 2

[0234] Preparation of Compound 4

[0235]

[0236] To a solution of 2,3-dihydroxy-2-methylpropanoic acid (4a, 11 mg, 0.090 mmol) in DMF (380 μl) was added (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ium methanesulfonate (1a, 200 μl, 0.070 - 0.08 mmol, in ~0.35 - 0.40 M in DMF, 3.0 eq. Et3N), HATU (34 mg, 0.090 mmol), and DIPEA (39 μl, 0.23 mmol). The resulting reaction mixture was stirred at room temperature for 1 hour. The mixture was then purified directly by reverse-phase column chromatography (20 - 60% MeCN / water, containing 0.1% formic acid modifier) to afford N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,3-dihydroxy-2-methylpropanamide (4, peak 2) as a solid. MS: m / z = 538 [M+H]. 1 1H NMR (500 MHz, DMSO-d6) δ 8.14 (d, J = 8.8 Hz, 1H), 7.77 (d, J = 10.9 Hz, 1H), 7.30 (s, 1H), 5.54–5.50 (m, 1H), 5.42 (s, 2H), 5.28 (d, J = 19.1 Hz, 1H), 5.16 (d, J = 19.1 Hz, 1H), 3.64 (d, J = 10.7 Hz, 1H), 3.39 (d, J = 10.7 Hz, 1H), 3.16 (br t, J = 6.0 Hz, 2H), 2.39 (s, 3H), 2.25–2.19 (m, 1H), 2.15–2.08 (m, 1H), 1.90–1.81 (m, 2H), 1.24 (s, 3H), 0.87 (t, J = 7.3 Hz, 3H).

[0237] The following compounds of the present disclosure in Table 2 were prepared using a similar method as described in Example 2 with different reaction times (10 minutes to 1 hour) and replacing the appropriate reactants and / or reagents:

[0238] Table 2

[0239]

[0240]

[0241]

[0242] Example 3

[0243] Preparation of Compound 21

[0244]

[0245] A solution of (R)-4-(difluoromethoxy)-2-hydroxybutyric acid (I-1c, 18 mg, 0.10 mmol), HATU (39 mg, 0.10 mmol) and N-methylmorpholine (0.020 mL, 0.18 mmol) in DMF (1 mL) was stirred at room temperature for about 10 minutes, then a mixture of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3,4:6,7]indazino[1,2-b]quinolin-1-ammonium methanesulfonate (1a, 0.25 mL, 0.089 - 0.10 mmol, ~0.35 - 0.40 M in DMF, containing 3.0 eq. Et3N) was added. The reaction mixture was stirred at room temperature for about 1 hour 40 minutes. The material was filtered and then subjected to reverse-phase column chromatography (25 - 70% MeCN / water, containing 0.1% formic acid modifier) to give (R)-4-(difluoromethoxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-hydroxybutyramide (21) as a solid. MS: m / z = 588 [M+H]. 11H NMR (500 MHz, DMSO-d6) δ 8.53 (d, J = 9.0 Hz, 1H), 7.79 (d, J = 10.7 Hz, 1H), 7.31 (s, 1H), 6.68 (t, J = 76.3 Hz, 1H, partially overlapping with the doublet at 6.51 ppm), 6.51 (d, J = 5.5 Hz, 1H, partially overlapping with the triplet at 6.68 ppm), 5.77 (d, J = 5.8 Hz, 1H), 5.61–5.54 (m, 1H), 5.42 (s, 2H), 5.25–5.12 (m, 2H), 4.11–4.05 (m, 1H), 4.01–3.94 (m, 2H), 2.40 (s, 3H), 2.23–2.11 (m, 3H), 2.03–1.92 (m, 2H), 1.92–1.81 (m, 2H), 0.87 (t, J = 7.2 Hz, 3H).

[0246] The following compounds of the present disclosure in Table 3 were prepared using a similar method as described in Example 3, where the reaction time was different (up to 4 hours) and appropriate reactants and / or reagents were substituted:

[0247] Table 3

[0248]

[0249] Example 4

[0250] Preparation of Compound 24

[0251]

[0252] To a solution of 3-hydroxy-2,2-dimethylpropanoic acid (24a, 22 mg, 0.19 mmol) in DMF (380 μl) was added DIPEA (99 μl, 0.56 mmol) and (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ium mesylate (1a, 250 μl, 0.089 - 0.10 mmol, ~0.35 - 0.40 M in DMF, containing 3.0 eq. Et3N), and then 1-propane phosphonic anhydride solution (in DMF) (150 μl, 0.25 mmol) was added. The resulting mixture was stirred overnight at room temperature and then purified by reverse-phase column chromatography (5 - 60% MeCN / water, containing 0.1% formic acid as modifier) to give N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2,2-dimethylpropanamide (24) as a solid. MS: m / z = 536 [M+H]. 1 1H NMR (500 MHz, DMSO-d6) δ 7.99 (d, J = 8.5 Hz, 1H), 7.79 (d, J = 11.0 Hz, 1H), 7.31 (s, 1H), 6.51 (s, 1H), 5.60–5.53 (m, 1H), 5.42 (s, 2H), 5.26–5.12 (m, 2H), 4.87 (t, J = 5.1 Hz, 1H), 3.45 (dd, J = 10.4, 5.0 Hz, 1H), 3.38 (dd, J = 10.5, 4.9 Hz, 1H), 3.18–3.12 (m, 2H), 2.40 (s, 3H), 2.20–2.06 (m, 2H), 1.92–1.80 (m, 2H), 1.11 (d, J = 9.8 Hz, 6H), 0.87 (t, J = 7.3 Hz, 3H).

[0253] The following compounds of the present disclosure in Table 4 were prepared using a method similar to that described in Example 4, where the reaction time was different (15 minutes to overnight) and appropriate reactants and / or reagents were substituted:

[0254] Table 4

[0255]

[0256]

[0257] Example 5

[0258] Preparation of Compound 28

[0259]

[0260] Step A - Compound 28b 1a Synthesis

[0261] To a solution of methyl (S)-2,2-dimethyl-1,3-dioxolane-4-carboxylate (28a, 0.15 g, 0.94 mmol) in DCE (3.0 mL) was added trimethyltin hydroxide (0.25 g, 1.4 mmol). The reaction mixture was stirred at 80 °C for 2.5 h. The mixture was cooled and diluted with EtOAc and aqueous dilute HCl (1.5 mmol). After shaking the mixture, the organic layer was separated, passed through a hydrophobic membrane phase separator, and concentrated under reduced pressure to give (S)-2,2-dimethyl-1,3-dioxolane-4-carboxylic acid (28b) as an oil, which was used directly in the next step without further purification.

[0262] Step B - Synthesis of Compound 28

[0263] To a solution of (S)-2,2-dimethyl-1,3-dioxolane-4-carboxylic acid (28b, 22 mg, 0.15 mmol) in DMF (1.0 mL) was added Hunig's base (0.050 mL, 0.29 mmol), followed by HATU (57 mg, 0.15 mmol). The reaction mixture was stirred at room temperature for 10 minutes, then a mixture of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl-1-ammonium mesylate (1a, 0.30 mL, 0.11 - 0.12 mmol, ~0.35 - 0.40 M in DMF, containing Hunig's base) was added. The reaction was stirred at room temperature for 1 hour 40 minutes, then another portion of (R)-2,2-dimethyl-1,3-dioxolane-4-carboxylic acid (30 mg, 0.21 mmol), HATU (70 mg, 0.18 mmol) and Hunig's base (0.050 mL, 0.29 mmol) in DMF (0.30 mL) was added. The reaction was stirred at room temperature for 20 minutes. Then, HCl (0.20 mL, 0.80 mmol, 4.0 M in dioxane) in water (0.20 mL) was added and the reaction was stirred at room temperature overnight. Another portion of HCl (0.20 mL, 4.0 M in dioxane) was added and the last portion of HCl (0.10 mL, 4.0 M in dioxane) was added 2 hours 20 minutes later. The reaction was stirred at room temperature for an additional 4.5 hours. The mixture was then filtered and subjected to reverse phase column chromatography (25 - 60% MeCN / water, containing 0.1% formic acid modifier) to afford (1S,9S)-1-((S)-2,3-dihydroxypropanamido)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-7-ium hexafluorophosphate (V) (28, peak 1, PF6 salt) as a solid. MS: m / z = 524 [M+H]. 11H NMR (500 MHz, DMSO-d6) δ 8.34 (d, J = 8.8 Hz, 1H), 7.79 (d, J = 10.9 Hz, 1H), 7.31 (s, 1H), 6.52 (s, 1H), 5.57–5.51 (m, 1H), 5.47 (d, J = 5.6 Hz, 1H), 5.42 (s, 2H), 5.22 (q, J = 18.9 Hz, 2H), 4.72 (t, J = 5.7 Hz, 1H), 4.03–3.99 (m, 1H), 3.66–3.60 (m, 1H), 3.57–3.51 (m, 1H), 3.23–3.10 (m, 2H), 2.40 (s, 3H), 2.27–2.19 (m, 1H), 2.12 (d, J = 4.7 Hz, 1H), 1.92–1.81 (m, 2H), 0.87 (t, J = 7.4 Hz, 3H).

[0264] The following compounds of the present disclosure in Table 5 were made using a method similar to that described in Example 5, substituting appropriate reactants and / or reagents:

[0265] Table 5

[0266]

[0267] Example 6

[0268] Preparation of Compound 30

[0269]

[0270] Step A - Synthesis of Compound 30b

[0271] A mixture of 2-(1-((benzyloxy)methyl)cyclopropyl)acetic acid (30a, 3.0 g, 14 mmol), MeOH (23 mL), and sulfuric acid (16 mL, 0.31 mmol) was stirred at room temperature for 3 days. The solvent was removed under reduced pressure. The residue was treated with DCM and water, then neutralized to pH 7 with sodium bicarbonate. The two layers were separated and the aqueous layer was washed twice more with DCM. The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure to give crude methyl 2-(1-((benzyloxy)methyl)cyclopropyl)acetate (30b) as an oil.

[0272] Step B - Synthesis of Compound 30c

[0273] At -78 °C, under argon, a solution of methyl 2-(1-((benzyloxy)methyl)cyclopropyl)acetate (30b, 300 mg, 1.3 mmol) in THF (4.0 mL) was added dropwise to a stirred solution of KHMDS (3.8 mL, 1.9 mmol, 0.5 M in toluene). The reaction mixture was stirred at -78 °C for 30 minutes, at which point a solution of 3-phenyl-2-(phenylsulfonyl)-1,2-oxaziridine (502 mg, 1.9 mmol) in THF (2.0 mL) was added dropwise. The reaction mixture was stirred at -78 °C for 30 minutes, then quenched with isopropanol and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 - 50% EtOAc / hexane) to give methyl 2-(1-((benzyloxy)methyl)cyclopropyl)-2-hydroxyacetate (30c).

[0274] Step C - Synthesis of Compound 30d

[0275] A mixture of methyl 2-(1-((benzyloxy)methyl)cyclopropyl)-2-hydroxyacetate (30c, 100 mg, 0.40 mmol), 10 wt% Pd / C (21 mg, 0.020 mmol Pd), and MeOH (2.0 mL) was stirred under H2 (1.0 atm) overnight. Then the reaction mixture was filtered through filter paper and rinsed with MeOH. The solvent was removed under reduced pressure to give crude 7-hydroxy-5-oxaspiro[2.4]heptan-6-one (30d). This crude mixture was used directly in the next step.

[0276] Step D - Synthesis of Compound 30e

[0277] A mixture of crude 7-hydroxy-5-oxaspiro[2.4]heptan-6-one (30d, 51 mg, 0.40 mmol), lithium hydroxide (9.6 mg, 0.40 mmol), and anhydrous MeOH (1.6 mL) was stirred at room temperature overnight. The solvent was removed under reduced pressure. The residue was treated with DMF (1.6 mL), imidazole (123 mg, 1.8 mmol), and tert-butyldimethylsilyl chloride (133 mg, 0.88 mmol). The reaction mixture was stirred at room temperature for 3 days. After removal of the solvent under reduced pressure, the mixture was diluted with EtOAc and washed with 1 M aqueous HCl. The aqueous layer was back-extracted twice with EtOAc. The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure to give crude 2-(1-(((tert-butyldimethylsilyl)oxy)methyl)cyclopropyl)-2-hydroxyacetic acid (30e) as an oil, which was used directly in the next step.

[0278] Step E - Synthesis of Compound 30f

[0279] To a stirred mixture of crude 2-(1-(((tert-butyldimethylsilyl)oxy)methyl)cyclopropyl)-2-hydroxyacetic acid (30e, 84 mg, 0.32 mmol), Hunig's base (73 μL, 0.42 mmol), (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ammonium methanesulfonate (1a, 380 μL, 0.13 - 0.15 mmol, in DMF ~0.35 - 0.40 M, containing 3.0 eq. Et3N) and DMF (0.30 mL) was added HATU (75 mg, 0.20 mmol). The reaction mixture was stirred at room temperature for 20 minutes, filtered through a syringe filter, and then subjected to reverse-phase column chromatography (10 - 95% MeCN / H2O, containing 0.1% formic acid modifier) to afford 2-(1-(((tert-butyldimethylsilyl)oxy)methyl)cyclopropyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-hydroxyacetamide (30f).

[0280] Synthesis of Compound 30 in Step F

[0281] A mixture of crude 2-(1-(((tert-butyldimethylsilyl)oxy)methyl)cyclopropyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-hydroxyacetamide (30f, 10 mg, 0.015 mmol), TFA (18 μL, 0.23 mmol), DMF (300 μL, 0.42 mmol) and MeOH (100 μL) was stirred at room temperature for 5 h. The resulting suspension was then dissolved in DMSO and subjected to reverse-phase column chromatography (10 - 100% MeCN / H2O, containing 0.1% formic acid modifier) to afford N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-hydroxy-2-(1-(hydroxymethyl)cyclopropyl)acetamide (30, a 1:1 mixture of diastereomers) as a solid. MS: m / z = 564 [M+H].1 1H NMR (500 MHz, DMSO-d6) δ 8.62 (d, J = 8.4 Hz, 1H), 8.30 (d, J = 8.8 Hz, 1H), 7.79 (apparent t, J = 11.7 Hz, 2H), 7.31 (s, 2H), 6.52 (s, 2H), 5.64–5.51 (m, 2H), 5.42 (s, 4H), 5.31–5.07 (m, 4H), 4.26 (d, J = 11.0 Hz, 1H), 3.90 (s, 1H), 3.77 (s, 1H), 3.44 (s, 2H, overlapping with water peak), 3.23–3.11 (m, 5H), 2.41 (s, 3H), 2.40 (s, 3H), 2.27–2.05 (m, 4H), 1.93–1.78 (m, 4H), 0.92–0.83 (m, 6H), 0.78–0.72 (m, 1H), 0.70–0.60 (m, 3H), 0.55–0.46 (m, 2H), 0.45–0.37 (m, 2H).

[0282] Example 7

[0283] Preparation of Compound 31

[0284]

[0285] Step A - Synthesis of Compound 31a

[0286] A mixture of (R)-3-hydroxydihydrofuran-2(3H)-one (1b, 600 mg, 5.9 mmol), methyl iodide (1.8 mL, 29 mmol), silver oxide (1.5 g, 6.6 mmol) and acetonitrile (12 mL) was stirred overnight at 75 °C under argon, in the dark. Then the mixture was filtered through filter, rinsed with acetonitrile, and the solvent was removed under reduced pressure. The residue was redissolved in DCM, refiltered, and concentrated under reduced pressure to give crude (R)-3-methoxydihydrofuran-2(3H)-one (31a) as an oil.

[0287] Step B - Synthesis of Compound 31b

[0288] A mixture of (R)-3-methoxydihydrofuran-2(3H)-one (31a, 350 mg, 3.0 mmol), lithium hydroxide (76 mg, 3.2 mmol) and methanol (6.0 mL) was stirred at room temperature for 3 days. Then the solvent was removed thoroughly under reduced pressure. To the resulting residue was added DMF (6.0 mL), imidazole (620 mg, 9.0 mmol) and tert-butyldimethylchlorosilane (590 mg, 3.9 mmol). The reaction mixture was stirred at room temperature overnight, at which time most of the solvent was removed under reduced pressure. The residue was diluted with DCM and washed with 1M aqueous HCl solution. The aqueous layer was back-extracted twice more with DCM. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure to give crude (R)-4-((tert-butyldimethylsilyl)oxy)-2-methoxybutyric acid (31b) as an oil, which was used directly in the next step.

[0289] Synthesis of Compound 31 in Step C

[0290] To a stirred mixture of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ammonium methanesulfonate (1a, 290 μL, 0.10 - 0.12 mmol, ~0.35 - 0.40 M in DMF, containing 3.0 eq. Et3N), crude (R)-4-((tert-butyldimethylsilyl)oxy)-2-methoxybutyric acid (31b, 140 mg, 0.57 mmol), Hunig's base (35 μL, 0.20 mmol) and DMF (0.10 mL) was added HATU (65 mg, 0.17 mmol). The mixture was stirred at room temperature for 10 minutes, at which time MeOH (0.10 mL) and TFA (77 μL, 1.0 mmol) were added. The mixture was stirred at room temperature for 2 hours. Then the resulting suspension was diluted with DMSO (0.30 mL) and subjected to reverse-phase column chromatography (10 - 70% MeCN / H2O, containing 0.1% formic acid as modifier) to give (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methoxybutyramide (31) as a solid. MS: m / z = 552 [M+H]. 11H NMR (500 MHz, DMSO-d6) δ 8.62 (d, J = 8.9 Hz, 1H), 7.75 (d, J = 10.9 Hz, 1H), 7.29 (s, 1H), 6.51 (s, 1H), 5.64–5.54 (m, 1H), 5.41 (s, 2H), 5.12 (s, 2H), 4.54 (s, 1H), 3.82 (dd, J = 8.5, 4.4 Hz, 1H), 3.51 (t, J = 6.3 Hz, 2H), 3.31 (s, 3H), 3.24–3.09 (m, 2H), 2.37 (s, 3H), 2.21–2.09 (m, 2H), 1.94–1.75 (m, 4H), 0.87 (t, J = 7.3 Hz, 3H).

[0291] Example 8

[0292] Preparation of Compound 32

[0293]

[0294] A mixture of methyl (S)-3-hydroxy-2-methylpropionate (32a, 17 mg, 0.14 mmol) and lithium hydroxide (3.4 mg, 0.14 mmol) was dissolved in DMF (190 μl) and stirred overnight at room temperature. Then, (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ammonium methanesulfonate (1a, 270 μl, 0.095 - 0.11 mmol, in DMF ~0.35 - 0.40 M, 3.0 eq. Et3N) was added, followed by HATU (43 mg, 0.11 mmol) and DIPEA (33 μl, 0.19 mmol). The resulting mixture was stirred at room temperature for 20 minutes and then purified by reverse-phase column chromatography (20 - 60% MeCN / water, containing 0.1% formic acid modifier) to afford (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2-methylpropanamide (32) as a solid. MS: m / z = 522 [M+H]. 11H NMR (500 MHz, DMSO-d6) δ 8.38 (d, J = 8.7 Hz, 1H), 7.77 (d, J = 11.0 Hz, 1H), 7.29 (s, 1H), 6.52 (s, 1H), 5.60–5.53 (m, 1H), 5.42 (s, 2H), 5.21 (s, 2H), 4.62 (s, 1H), 3.61–3.51 (m, 1H), 3.36–3.34 (m, 1H), 3.22–3.10 (m, 2H), 2.47–2.40 (m, 1H), 2.38 (s, 3H), 2.20–2.05 (m, 2H), 1.94–1.79 (m, 2H), 1.01 (d, J = 6.9 Hz, 3H), 0.87 (t, J = 7.3 Hz, 3H).

[0295] The following compounds of the present disclosure in Table 6 were prepared using a method similar to that described in Example 8 with different reaction times (2 hours for the LiOH phase and 15 - 30 minutes for the coupling phase) and replacing the appropriate reactants and / or reagents:

[0296] Table 6

[0297]

[0298]

[0299] Example 9

[0300] Preparation of Compound 38

[0301]

[0302] Step A - Synthesis of Compound 38b

[0303] Ethanol (2.0 mL) was added to a mixture of 3,3 - dimethyldihydrofuran - 2(3H) - one (38a, 0.068 g, 0.60 mmol) and lithium hydroxide (0.014 g, 0.60 mmol). The reaction mixture was heated to 60 °C overnight. The reaction mixture was cooled and concentrated under reduced pressure. The resulting crude product, lithium 4 - hydroxy - 2,2 - dimethylbutyrate (38b), which was a solid, was used directly in the next step without further purification.

[0304] Step B - Synthesis of Compound 38c

[0305] To a mixture of lithium 4-hydroxy-2,2-dimethylbutyrate (38b, 0.079 g, 0.60 mmol), DMAP (7.0 mg, 0.060 mmol), and TBSCl (0.10 g, 0.66 mmol) in DMF (3.0 mL) was added TEA (0.10 mL, 0.72 mmol). The reaction mixture was stirred overnight at room temperature. The mixture was diluted with DCM, water, and 1 M HCl (1.5 mL). The mixture was extracted and passed through a hydrophobic membrane phase separator. The organic layer was concentrated under reduced pressure, and the resulting crude 4-((tert-butyldimethylsilyl)oxy)-2,2-dimethylbutyric acid (38c) was used directly in the next step.

[0306] Step C - Synthesis of Compound 38

[0307] To a solution of 4-((tert-butyldimethylsilyl)oxy)-2,2-dimethylbutyric acid (38c, 150 mg, 0.60 mmol) and N-methylmorpholine (0.050 mL, 0.46 mmol) in DMF (1.0 mL) was added HATU (170 mg, 0.45 mmol). The reaction mixture was stirred at room temperature for 5 minutes, then a mixture of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ammonium methanesulfonate (1a, 0.30 mL, 0.11 - 0.12 mmol, ~0.35 - 0.40 M in DMF, containing 3.0 eq. Et3N) was added. The reaction mixture was stirred at room temperature for 2 hours 40 minutes. Then, MeOH (0.15 mL) was added, followed by TFA (0.20 mL). The reaction mixture was stirred at room temperature for 1 hour. Then TEA (0.30 mL) was added, the mixture was filtered and subjected to reverse-phase column chromatography (25 - 65% MeCN / water, containing 0.1% formic acid modifier) to give N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2,2-dimethylbutyramide (38) as a solid. MS: m / z = 550 [M+H]. 11H NMR (500 MHz, DMSO-d6): δ 8.07 (d, J = 8.4 Hz, 1H), 7.79 (d, J = 10.9 Hz, 1H), 7.31 (s, 1H), 5.60–5.54 (m, 1H), 5.42 (s, 2H), 5.22–5.08 (m, 2H), 3.46–3.41 (m, 4H) (signals overlapped with broad water peak), 3.20–3.11 (m, 3H) (signals overlapped with broad water peak), 2.40 (s, 3H), 2.16–2.06 (m, 2H), 1.90–1.81 (m, 2H), 1.72 (t, J = 7.4 Hz, 2H), 1.17 (d, J = 6.6 Hz, 6H), 0.88 (t, J = 7.3 Hz, 3H).

[0308] Example 10

[0309] Preparation of Compounds 39 and 40

[0310]

[0311] Step A - Synthesis of Compound 39b

[0312] At room temperature, dihydrofuran-2(3H)-one (39a, 10 g, 116 mmol) and ethyl formate (8.6 g, 120 mmol) were added dropwise to a stirred mixture of NaH (4.7 g, 120 mmol, 60 wt%) in hexane (100 mL). After adding about 10% of the mixture, a small amount of absolute ethanol (0.70 mL) was added to initiate the reaction, and then the mixture was refluxed at 70 °C for 2 hours. The mixture was filtered, and the filter cake was washed with hexane (60 mL) and dried in vacuo to give crude sodium (Z)-(2-oxodihydrofuran-3(2H)-ylidene)methanolate (39b) as a solid, which was used directly in the next step of the reaction.

[0313] Step B - Synthesis of Compound 39c

[0314] Sodium (Z)-(2-oxodihydrofuran-3(2H)-ylidene)methanolate (39b, 15 g, 110 mmol) (freshly prepared) in CH3I (43 mL) in a round-bottom flask equipped with a reflux condenser was heated to reflux (60 °C) under a nitrogen atmosphere for 48 hours. The reaction mixture was cooled to room temperature and filtered. The flask and the filter residue were washed with chloroform (CHCl3), and the combined filtrate was evaporated under reduced pressure. The filtrate was purified by silica gel column chromatography (1:1 petroleum ether:EtOAc) to give 3-methyl-2-oxotetrahydrofuran-3-carbaldehyde (39c) as an oil.

[0315] Step C - Synthesis of Compound 39d

[0316] DAST (19 mL, 140 mmol) was added dropwise to a solution of 3-methyl-2-oxotetrahydrofuran-3-carbaldehyde (39c, 6.0 g, 47 mmol) in DCM (60 mL) at 0 °C. The reaction mixture was stirred at room temperature for 18 h. The mixture was added to saturated aqueous NaHCO3 (300 mL), then adjusted to pH 8, and the mixture was extracted with DCM (3 × 60 mL). The combined organic fractions were washed with brine (20 mL), dried over Na2SO4, filtered, and the solvent was evaporated under reduced pressure to give crude 3-(difluoromethyl)-3-methyldihydrofuran-2(3H)-one (39d) as an oil. This material was used directly in the next step.

[0317] Step D - Synthesis of compound 39e

[0318] At room temperature, LiOH (0.96 g, 40 mmol) was added to a stirred mixture of 3-(difluoromethyl)-3-methyldihydrofuran-2(3H)-one (39d, 2.0 g, 13 mmol) in MeOH / THF / H2O (1:1:1) (10 mL), and the mixture was stirred at room temperature for 18 h. The solvent was evaporated under reduced pressure to give crude lithium 2-(difluoromethyl)-4-hydroxy-2-methylbutanoate (39e) as a solid, which was used directly in the next step.

[0319] Step E - Synthesis of compound 39f

[0320] At room temperature, TBSCl (0.43 g, 2.9 mmol) was added to a stirred mixture of imidazole (0.32 g, 4.8 mmol) and lithium 2-(difluoromethyl)-4-hydroxy-2-methylbutanoate (39e, 0.40 g, 2.4 mmol) in DMF (6.0 mL), and the mixture was stirred at room temperature for 18 h. The material was purified by silica gel column chromatography (10% EtOAc / petroleum ether) to give 4-((tert-butyldimethylsilyl)oxy)-2-(difluoromethyl)-2-methylbutanoic acid (39f) as an oil, which was used directly in the next step.

[0321] Step F - Synthesis of compounds 39g and 39h

[0322] At room temperature, DIPEA (0.10 mL, 0.57 mmol) and HATU (86 mg, 0.23 mmol) were added to a stirred mixture of 4-((tert-butyldimethylsilyl)oxy)-2-(difluoromethyl)-2-methylbutanoic acid (39f, 500 mg, 1.8 mmol) in DMF (2.0 mL). The mixture was stirred for 5 minutes and then (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3,4:6,7]indazino[1,2-b]quinolin-1-ium methanesulfonate (1a, 100 mg, 0.19 mmol) was added. The mixture was stirred at room temperature for 30 minutes. The material was purified by reverse-phase column chromatography (75 - 100% MeCN / water, containing 0.1% formic acid modifier) to afford 4-((tert-butyldimethylsilyl)oxy)-2-(difluoromethyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)butanamide (39g, peak 1) as a solid, and 4-((tert-butyldimethylsilyl)oxy)-2-(difluoromethyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)butanamide (39h, peak 2) as a solid. MS: m / z = 700 [M+H].

[0323] Synthesis of Compounds 39 and 40, Step G

[0324] At room temperature, TFA (0.050 mL, 0.65 mmol) was added to a stirred mixture of 4-((tert-butyldimethylsilyl)oxy)-2-(difluoromethyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-methylbutanamide (40 mg, 0.057 mmol, 39 g, peak 1) in DMF / MeOH (0.75 mL), and the mixture was stirred at room temperature for 1 hour. Then, more TFA (0.10 mL) was added, and the mixture was stirred at room temperature for 0.5 hour. The residue was purified by reverse-phase column chromatography (30 - 60% MeCN / water, containing 0.1% formic acid modifier), and then again by reverse-phase column chromatography (26 - 56% MeCN / water, containing 0.1% formic acid modifier) to afford 2-(difluoromethyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methylbutanamide (40) as a solid. MS: m / z = 586 [M+H]. 1 H NMR (DMSO-d6, 400 MHz) δ 8.36 (br d, 1H, J = 8.1 Hz), 7.81 (d, 1H, J = 11.0 Hz), 7.32 (s, 1H), 6.52 (s, 1H), 6.1–6.5 (m, 1H), 5.6–5.7 (m, 1H), 5.42 (s, 2H), 5.2–5.3 (m, 1H), 5.05 (br d, 1H, J = 18.8 Hz), 4.65 (t, 1H, J = 4.8 Hz), 3.40–3.70 (m, 2H), 3.11–3.20 (m, 2H), 2.41 (s, 3H), 2.01–2.21 (m, 2H), 1.84–1.88 (m, 3H), 1.60–1.71 (m, 1H), 1.22 (s, 3H), 0.88 (brt, 3H, J = 7.3 Hz).

[0325] At room temperature, TFA (0.050 ml, 0.65 mmol) was added to a stirred mixture of 4-((tert-butyldimethylsilyl)oxy)-2-(difluoromethyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-methylbutanamide (39h, 45 mg, 0.064 mmol, peak 2), and the mixture was stirred at room temperature for 1 hour. The residue was purified by reverse-phase column chromatography (30 - 60% MeCN / water, containing 0.1% formic acid modifier) to give 2-(difluoromethyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methylbutanamide (39) as a solid. MS: m / z = 586 [M+H]. 1 1H NMR (DMSO-d6, 400 MHz) δ 8.42 (d, 1H, J = 8.1 Hz), 7.86 (d, 1H, J = 11.0 Hz), 7.36 (s, 1H), 6.59 (s, 1H), 6.27 (br t, 1H, J = 56.3 Hz), 5.6–5.8 (m, 1H), 5.47 (s, 2H), 5.3–5.4 (m, 1H), 5.07 (d, 1H, J = 18.8 Hz), 4.70 (t, 1H, J = 4.9 Hz), 3.5–3.6 (m, 2H), 3.1–3.3 (m, 2H), 2.45 (s, 3H), 2.07–2.17 (m, 2H), 1.84–1.93 (m, 3H), 1.61–1.67 (m, 1H), 1.27 (s, 3H), 0.87 (t, 3H, J = 7.2 Hz).

[0326] Example 11

[0327] Preparation of Compound 41

[0328]

[0329] Step A - Synthesis of Compound 41b

[0330] To a stirred mixture of 2-((tert-butoxycarbonyl)amino)-2-methylpropanoic acid (41a, 31 mg, 0.15 mmol), Hunig's base (35 μL, 0.20 mmol), (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ium methanesulfonate (1a, 290 μL, 0.10 - 0.12 mmol, ~0.35 - 0.40 M in DMF, containing 3 eq. Et3N) and DMF (0.20 mL) was added HATU (57 mg, 0.15 mmol). The reaction mixture was stirred at room temperature for 10 minutes, filtered through a syringe filter, and then subjected to reverse-phase column chromatography (20 - 100% MeCN / H2O, containing 0.1% formic acid modifier) to afford tert-butyl (1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-2-methyl-1-oxopropan-2-yl)carbamate (41b) as a solid. MS: m / z = 621 [M+H].

[0331] Step B - Synthesis of Compound 41

[0332] A mixture of tert-butyl (1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-2-methyl-1-oxopropan-2-yl)carbamate (41b, 58 mg, 0.094 mmol), 4 M HCl in dioxane (0.35 mL, 1.4 mmol) and dioxane (0.65 mL) was stirred at room temperature for 2.5 hours and monitored by LCMS. The mixture was then diluted with H2O and subjected to reverse-phase column chromatography (10 - 70% MeCN / H2O, containing 0.1% formic acid modifier) to afford 2-amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-methylpropanamide formate (41) as a solid. MS: m / z = 521 [M+H]. 11H NMR (500 MHz, DMSO-d6) δ 8.55 (s, 1H), 8.27 (s, 1H), 7.78 (d, J = 10.9 Hz, 1H), 7.30 (s, 1H), 5.53 (t, J = 5.2 Hz, 1H), 5.41 (s, 2H), 5.16 (s, 2H), 3.16 (t, J = 6.1 Hz, 2H), 2.39 (s, 3H), 2.22–2.08 (m, 2H), 1.93–1.78 (m, 2H), 1.37 (s, 3H), 1.33 (s, 3H), 0.87 (t, J = 7.3 Hz, 3H).

[0333] The following compounds of the present disclosure in Table 7 were prepared using a similar method as described in Example 11, where the reaction times were different (10 - 30 minutes for Step A and 3 - 7 hours for Step B) and appropriate reactants and / or reagents were replaced:

[0334] Table 7

[0335]

[0336] Example 12

[0337] Preparation of Compound 45

[0338]

[0339] Step A - Synthesis of Compound 45

[0340] To a stirred mixture of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ium methanesulfonate (1a, 270 μL, 0.095 - 0.11 mmol, ~0.35 - 0.40 M in DMF, containing 3 eq. Et3N), (((9H-fluoren-9-yl)methoxy)carbonyl)-L-serine (45a, 39 mg, 0.12 mmol), Hunig's base (18 μL, 0.10 mmol) and DMF (270 μL) was added HATU (46 mg, 0.12 mmol). The reaction mixture was stirred at room temperature for 5 minutes, at which point 4-methylpiperidine (59 μL, 0.50 mmol) was added. The mixture was stirred at room temperature for an additional 45 minutes and monitored by LCMS. Then, the reaction mixture was subjected to reverse phase column chromatography (10 - 60% MeCN / H2O, containing 0.1% formic acid modifier) to afford (S)-2-amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide formate (45) as a solid. MS: m / z = 523 [M+H]. 1 1H NMR (500 MHz, DMSO-d6) δ 8.70 (s, 1H), 8.23 (s, 1H), 7.79 (d, J = 10.9 Hz, 1H), 7.30 (s, 1H), 5.57 (broad s, 1H), 5.42 (s, 2H), 5.27–5.14 (m, 2H), 3.63–3.44 (m, 3H), 3.24–3.08 (m, 2H), 2.39 (s, 3H), 2.26–2.18 (m, 1H), 2.17–2.05 (m, 1H), 1.97–1.76 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0341] The following compounds of the present disclosure in Table 8 were prepared using a similar method as described in Example 12 with different reaction times (coupling stage from 5 minutes to 2 hours, Fmoc deprotection stage from 30 minutes to 14 hours and up to 40 °C) and substituting appropriate reactants and / or reagents:

[0342] Table 8

[0343]

[0344] Example 13

[0345] Preparation of Compound 49

[0346]

[0347] Step - A - Synthesis of Compound 49b

[0348] To a mixture of 3,3 - difluoropyrrolidin - 2 - one (49a, 1.6 g, 13 mmol), Boc - anhydride (6.1 mL, 26 mmol) and DMF (13 mL) was added DMAP (0.16 g, 1.3 mmol). The reaction mixture was stirred overnight at room temperature. Then the mixture was diluted with DCM, washed three times with brine, dried over MgSO4, filtered and concentrated under reduced pressure to give crude tert - butyl 3,3 - difluoro - 2 - oxopyrrolidine - 1 - carboxylate (49b) as an oil, which was used directly in the next step.

[0349] Step B - Synthesis of Compound 49c

[0350] A mixture of crude tert - butyl 3,3 - difluoro - 2 - oxopyrrolidine - 1 - carboxylate (49b, 181 mg, 0.82 mmol), lithium hydroxide (9.6 mg, 0.40 mmol) and DMF (0.40 mL) was stirred at 60 °C for 30 minutes and then stirred overnight at room temperature. To the resulting suspension was added (1S,9S) - 9 - ethyl - 5 - fluoro - 9 - hydroxy - 4 - methyl - 10,13 - dioxo - 2,3,9,10,13,15 - hexahydro - 1H,12H - benzo[de]pyrano[3',4':6,7]indazino[1,2 - b]quinolin - 1 - ammonium methanesulfonate (1a, 540 μL, 0.19 - 0.22 mmol, in DMF ∼0.35 - 0.40 M, containing 3.0 eq. Et3N) and HATU (140 mg, 0.36 mmol). The mixture was stirred at room temperature for 20 minutes. Then, the mixture was dissolved in DMSO (1.0 mL) and subjected to reverse - phase column chromatography (10 - 100% MeCN / H2O, containing 0.1% formic acid modifier) to give (4 - ((((1S,9S) - 9 - ethyl - 5 - fluoro - 9 - hydroxy - 4 - methyl - 10,13 - dioxo - 2,3,9,10,13,15 - hexahydro - 1H,12H - benzo[de]pyrano[3',4':6,7]indazino[1,2 - b]quinolin - 1 - yl)amino) - 3,3 - difluoro - 4 - oxobutyl)carbamic acid tert - butyl ester (49c) as a solid.

[0351] Step C - Synthesis of Compound 49

[0352] A mixture of tert-butyl (4-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3,3-difluoro-4-oxobutyl)carbamate (49c, 30 mg, 0.045 mmol), TFA (35 μL, 0.45 mmol) and DCM (0.30 mL) was stirred at room temperature for 5 h. The mixture was then diluted with DMSO and subjected to reverse-phase column chromatography (10 - 70% MeCN / H2O, containing 0.1% formic acid modifier) to afford 4-amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,2-difluorobutanamide (49) as a solid. MS: m / z = 557 [M+H]. 1 1H NMR (500 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.81 (d, J = 10.9 Hz, 1H), 7.32 (s, 1H), 6.54 (br s, 1H), 5.60 (t, J = 5.5 Hz, 1H), 5.42 (s, 2H), 5.25 (d, J = 18.6 Hz, 1H), 5.10 (d, J = 18.6 Hz, 1H), 3.24–3.11 (m, 2H), 2.81 (t, J = 6.9 Hz, 2H), 2.40 (s, 3H), 2.38–2.26 (m, 2H), 2.25–2.13 (m, 2H), 1.96–1.79 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0353] Example 14

[0354] Preparation of Compound 50

[0355]

[0356] To a stirred mixture of (S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-((tert-butoxycarbonyl)amino)propanoic acid (50a, 51 mg, 0.12 mmol), Hunig's base (21 μL, 0.12 mmol), (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ium mesylate (1a, 290 μL, 0.10 - 0.12 mmol, ~0.35 - 0.40 M in DMF, containing 3.0 eq. Et3N) and DMF (0.30 mL) was added HATU (46 mg, 0.12 mmol). The reaction mixture was stirred at room temperature for 20 min, at which point 4-methylpiperidine (24 μL, 0.20 mmol) was added. After stirring at 40 °C for 2 h, the reaction mixture was subjected to reverse phase column chromatography (10 - 50% MeCN / H2O, containing 0.1% formic acid modifier) to afford ((S)-2-amino-3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3-oxopropyl)carbamic acid tert-butyl ester (50b) as a solid.

[0357] Step B - Synthesis of Compound 50

[0358] A mixture of tert-butyl ((S)-2-amino-3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3-oxopropyl)carbamate (50b, 44 mg, 0.071 mmol), TFA (0.11 mL, 1.4 mmol) and DCM (0.30 mL) was stirred at room temperature for 45 minutes. The mixture was then quenched with N-methylmorpholine (0.16 mL, 1.4 mmol), diluted with water and subjected to reverse phase column chromatography (10 - 50% MeCN / H2O, containing 0.1% formic acid modifier) to afford (S)-2,3-diamino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-propanamide formate (50) as a solid. MS: m / z = 522 [M+H]. 1 1H NMR (500 MHz, DMSO-d6) δ 8.65 (br s, 1H), 8.14 (s, 1H), 7.81 (d, J = 10.9 Hz, 1H), 7.32 (s, 1H), 6.67 (br s, 3H), 5.59–5.50 (m, 1H), 5.42 (s, 2H), 5.26 (d, J = 18.7 Hz, 1H), 5.20 (d, J = 18.8 Hz, 1H), 3.48 (dd, J = 8.9, 4.6 Hz, 1H), 3.23–3.16 (m, 2H), 3.08 (dd, J = 12.7, 4.6 Hz, 1H), 2.81 (dd, J = 12.7, 9.0 Hz, 1H), 2.41 (s, 3H), 2.27–2.18 (m, 1H), 2.18–2.07 (m, 1H), 1.93–1.78 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0359] Example 15

[0360] Preparation of Compound 51

[0361]

[0362] Step A - Preparation of Compound 51

[0363] A mixture of (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-ammonium mesylate (1a, 270 μL, 0.095 - 0.11 mmol, ~0.35 - 0.40 M in DMF, containing 3.0 eq. TEA), iodoethanol (16 μL, 0.020 mmol), DIPEA (35 μL, 0.20 mmol) and DMF (0.20 mL) was stirred at 40 °C for 3 days. The mixture was then purified directly by reverse-phase column chromatography (10 - 90% MeCN / water, containing 0.1% formic acid modifier) to give (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((2-hydroxyethyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione formate (51). MS: m / z = 480 [M+H]. 1 1H NMR (500 MHz, DMSO-d6) δ 8.13 (s, 1H), 7.73 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.51 (s, 1H), 5.45–5.31 (m, 4H), 4.33 (s, 1H), 3.58–3.48 (m, 2H), 3.24–3.13 (m, 1H), 3.02 (dt, J = 17.2, 5.0 Hz, 1H), 2.88 (dt, J = 11.6, 6.2 Hz, 1H), 2.77 (dt, J = 11.5, 5.6 Hz, 1H), 2.37 (s, 3H), 2.32–2.23 (m, 1H), 2.11–2.00 (m, 1H), 1.95–1.78 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0364] The following compounds of the present disclosure in Table 9 were prepared using a similar method as described in Example 15 with different reaction times (2 hours to 1 week) and temperatures (23 - 50 °C) and by replacing the appropriate alkyl electrophiles:

[0365] Table 9

[0366]

[0367]

[0368]

[0369] Example 16

[0370] Preparation of Compound 60

[0371]

[0372] Step A - Synthesis of Compound 60

[0373] At 0 °C, formaldehyde (37 wt% in water, 26 μL, 0.35 mmol), acetic acid (4.0 μL, 0.070 mmol), and sodium triacetoxyborohydride (15 mg, 0.070 mmol) were added to a suspension of (1S,9S)-1-((2,2-difluoroethyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione (I-60a, 17 mg, 0.035 mmol) in MeOH (0.70 mL). After 6 hours, DCM (0.60 mL) was added to dissolve the mixture, and then formaldehyde (37 wt% in water, 26 μL, 0.35 mmol), acetic acid (4.0 μL, 0.070 mmol), and sodium triacetoxyborohydride (15 mg, 0.070 mmol) were added. The mixture was stirred at room temperature for 3 days. At this time, formaldehyde (37 wt% in water, 52 μL, 0.70 mmol), acetic acid (8.0 μL, 0.14 mmol), and sodium triacetoxyborohydride (29 mg, 0.14 mmol) were added, and the mixture was stirred at room temperature for 30 minutes. This step was repeated once more. Then the mixture was subjected to reverse-phase column chromatography (30 - 90% MeCN / H2O, containing 0.1% formic acid modifier) to obtain (1S,9S)-1-((2,2-difluoroethyl)(methyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione as a solid. MS: m / z = 514 [M+H]. 1HNMR(500MHz,DMSO-d6)δ7.75(d,J=10.8Hz,1H),7.31(s,1H),6.49(s,1H),6.29(tt,J=56.3,4.1Hz,1H),5.42(s,2H),5.37(d,J=19.6Hz,1H),5.27(d,J=19.8Hz,1H),4.45(dd,J=10.9,4.6Hz,1H),3.07–2.85(m,2H),2.37(s,3H),2.34–2.22(m,4H),2.11–1.99(m,1H),1.94–1.79(m,2H),0.88(t,J=7.3Hz,3H).

[0374] Example 17

[0375] CellTiter- 2.0 Cytotoxicity Assay

[0376] Step 1: Seed a 384-well plate for the assay on Day 0 (45 μL per well).

[0377] Quickly thaw the Jeko-1 cells in the cryovial by incubating them in a 37 °C water bath for less than 1 minute until only a small amount of ice remains in the vial. Immediately remove the vial and wipe it with 70% ethanol. Transfer the cells from the vial to a sterile centrifuge tube containing 8 mL of pre-warmed cell medium. Rinse the vial with an additional 1 mL of medium to ensure complete transfer of the cells to the centrifuge tube. Then centrifuge the cells at 150 x g for 5 minutes. Aspirate the supernatant and resuspend the cell pellet in 10 - 20 mL of cell medium. Count the cells using Vi-cell and prepare 6.6×10 4 cells / ml, 3000 cells / 45 μL / well. Then add 45 μL / well of cells to a 384-well low-edge white flat-bottom polystyrene TC-treated microplate (Corning, Cat# 3570) (if needed, dispense 1 blank plate in 20 μL to help standardize the Combi, using medium speed). Spin the plate at 150 x g for 30 seconds.

[0378] Step 2: Add the compound on Day 1

[0379] Retrieve the compound plate and the reference compound stock solution and thaw at room temperature. Centrifuge the tubes at 2000 x g for 30 seconds. Prepare a 10X intermediate assay plate (Greiner plate, Cat# 781280) using an Echo liquid handler. Prepare serial dilutions using the appropriate buffer (HBSS (Gibco, Cat# 14025 - 092) + 10 mM HEPES (Gibco, Cat# 15630 - 080) + 0.1% BSA (Sigma, Cat# A9576)). Max_E uses medium (cell-free). Transfer the compounds (5 μL 10X) from the intermediate plate to the detection plate using a Bravo liquid handler very slowly so as not to disrupt the cell monolayer. Spin the plate at 150 x g for 30 seconds.

[0380] Step 3: Perform a CellTiter - Glo 2.0 assay (Promega, Cat# G9242) on Day 4 or Day 5 (the CellTiter - Glo kit is stored at -70 °C)

[0381] Thaw the CellTiter - 2.0 reagent overnight at 4 °C, taking care not to expose the reagent to temperatures above 25 °C. Equilibrate the kit at room temperature for approximately 30 minutes. Using a Standard Cassette Combi, add the CellTiter - 2.0 reagent (20 μl) to 50 μl of medium containing cells. Mix the contents on an orbital shaker for 2 - 3 minutes to induce cell lysis. Spin the plate at 150 x g for 30 seconds. Incubate the plate at room temperature for 5 minutes to stabilize the luminescence signal. Record the luminescence to calculate the EC 50 value, using an integration time of 0.25 - 1 second per well as a guide.

[0382] The exemplary compounds of the present disclosure were tested in the assay of Example 15 above, and the results are provided in Table 9 below:

[0383] Table 9

[0384]

[0385]

[0386]

Claims

1. A compound of structural formula I or a pharmaceutically acceptable salt or solvate thereof: Wherein: R k selected from hydrogen, -C 1-6 alkyl, (CH2) n C(O)NHC 1-6 alkyl, (CH2) n C 6-10 aryl and said alkyl and aryl are optionally substituted by 1 to 3 hydroxy groups, -C 1-6 alkyl OH groups, and said alkyl is further optionally substituted by 1 to 10 halogen groups; R j represents hydrogen or C 1-6 alkyl, said alkyl being optionally substituted by 1 to 10 halogens; R 2 and R 3 are independently selected from hydrogen, -C 1-6 alkyl, OH, -C 1-6 alkylOH, halogen, -C 1-9 haloalkyl, -(CH2) n NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -C 3-6 cycloalkyl, -(CH2) n C 6-10 aryl and -(CH2) n OC 1-6 alkyl, provided that R 2 and R 3 are not both halogen; R 4 Selected from C 1-6 alkyl, OH, -C 1-6 alkyl OH, -CH(OH)C 1-6 alkyl, -C 1-9 haloalkyl, halogen, -C 3-6 cycloalkyl, -(CH2) n OC 1-3 alkyl, -(CH2) n OC 1-9 haloalkyl, -CR x R y C 1-6 alkyl OH, -(CH2) n NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2 and -NHC(O)C 1-6 alkyl NH2; R x and R y represent a combination to form C 3-6 an alkylene group of a cycloalkyl or spirocycloalkyl; 1-3 an alkylene group; Each n independently represents 0, 1, 2 or 3.

2. The compound according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein any hydrogen atom in the alkyl group of R 2 , R 3 and / or R 4 is not deuterated.

3. The compound according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein R 2 is selected from hydrogen, C 1-6 alkyl, CH2OC 1-6 alkyl, -CH2OH, -O(CH2)2OH, -CH2F, -CHF2, -CF3, -(CH2)phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -(CH2) n NH2, -NHCH3 and -N(CH3)2.

4. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt or solvate thereof, wherein R 2 is selected from hydrogen, C 1-6 alkyl, (CH2) n OCH3, -OH, -CH2OH, -CH2F, -CHF2, -CF3, and -NH2.

5. The compound according to any one of claims 1-4, or a pharmaceutically acceptable salt or solvate thereof, wherein R 3 is selected from hydrogen, C 1-6 alkyl, CH2OC 1-6 alkyl, -CH2OH, -(CH2)2OH, -CH2F, -CHF2, -CF3, -(CH2)phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -(CH2) n NH2, -NHCH3 and -N(CH3)2.

6. The compound according to any one of claims 1-5, or a pharmaceutically acceptable salt or solvate thereof, wherein R 3 is selected from hydrogen, C 1-6 alkyl, (CH2) n OCH3, -OH, -CH2OH, -CH2F, -CHF2, -CF3, and -NH2.

7. The compound according to any one of claims 1-6, or a pharmaceutically acceptable salt or solvate thereof, wherein R 2 and R 3 One of them is selected from -OH, -CH2OH, -(CH2)2OH and (CH2) n NH2, and the other is selected from hydrogen, CH2 phenyl and C 1-6 alkyl.

8. The compound according to any one of claims 1-7, or a pharmaceutically acceptable salt or solvate thereof, wherein R 4 is selected from CH3, OH, -CH2OH, -(CH2)2OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3, (CH2) n OCHF2, cyclopropyl, -(CH2) n NH2 and -cyclopropylCH2OH, -NHC(O)CH(CH3)NH2.

9. The compound according to claims 1-8 or a pharmaceutically acceptable salt or solvate thereof, wherein R 4 is OH, -(CH2) n OH, -CH(CH3)OH, -(CH2) n NH2, or R 4 is -CR x R y C 1-6 alkyl OH, wherein R x and R y are alkylene substituents that combine to form cyclopropyl.

10. The compound according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein R 2 and R 3 one of which is -OH or -C 1-6 alkyl OH, and the other is selected from hydrogen, CH2 phenyl and C 1-6 alkyl, and R 4 is selected from CH3, OH, -C 1-6 alkyl OH, -C 1-3 haloalkyl, -CR x R y C 1-6 alkyl OH, -(CH2) n NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2 and -NHC(O)C 1-6 alkyl NH2.

11. The compound according to claim 10, or a pharmaceutically acceptable salt or solvate thereof, wherein one of R 2 and R 3 is -OH or -C 1-6 alkyl OH, and the other is selected from hydrogen, CH2 phenyl, and C 1-6 alkyl, and R 4 is selected from -CH2OH, -(CH2)2OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3, and -(CH2) n NH2.

12. The compound according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein R 2 and R 3 One of them is -(CH2) n NH2, -NHCH3 or -N(CH3)2, and the other is selected from hydrogen, CH2 phenyl and C 1-6 alkyl, and R 4 is selected from CH3, OH, -CH2OH, -(CH2)2OH, -C 1-3 haloalkyl, -CR x R y C 1-6 alkyl OH, -(CH2) n NH2, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2 and -NHC(O)C 1-6 alkyl NH2.

13. The compound according to claim 12, or a pharmaceutically acceptable salt or solvate thereof, wherein R 2 and R 3 one of which is (CH2) n NH2, and the other is selected from hydrogen, CH2 phenyl and C 1-6 alkyl, and R 4 is selected from -CH2OH, -(CH2)2OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3 and -(CH2) n NH2.

14. The compound or a pharmaceutically acceptable salt or solvate thereof according to claims 1-13, which is represented by structural formula II: wherein R 4 is selected from -CH2OH, -(CH2)2OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3 and -(CH2) n NH2 15. The compound or a pharmaceutically acceptable salt or solvate thereof according to claims 1-13, which is represented by structural formula III: wherein R 4 is selected from CH3, -CH2OH, -(CH2)2OH, -CH(CH3)OH, -CH2F, -CHF2, -CF3 and -(CH2) n NH2 16. The compound or a pharmaceutically acceptable salt or solvate thereof according to claims 1-13, which is represented by structural formula IV: wherein R 4 is -CH2F, -CHF2, -CF3 or -(CH2) n NH2 17. A compound or a pharmaceutically acceptable salt or solvate thereof, which is: N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,4-dihydroxybutyramide; (R)-N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,4-dihydroxybutyramide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,4-dihydroxybutyramide; (S)-N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,4-dihydroxybutyramide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,4-dihydroxy-3,3-dimethylbutyramide; (R)-N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,4-dihydroxy-3,3-dimethylbutyramide; (S)-N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,4-dihydroxy-3,3-dimethylbutyramide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,3-dihydroxy-2-methylpropanamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,3-dihydroxy-2-methylpropanamide; N-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,3-dihydroxy-2-methylpropanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,3-dihydroxy-2-methylpropanamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,3-dihydroxy-2-methylpropanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2-(hydroxymethyl)-2-methylpropanamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2-(hydroxymethyl)-2-methylpropanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,2-difluoro-3-hydroxybutyramide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,2-difluoro-3-hydroxybutyramide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,2-difluoro-3-hydroxybutyramide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,2-difluoro-3-hydroxybutyramide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,2-difluoro-3-hydroxypropanamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,2-difluoro-3-hydroxypropanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoro-2-(hydroxymethyl)acrylamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoro-2-(hydroxymethyl)acrylamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoro-2-(hydroxymethyl)acrylamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoro-2-(hydroxymethyl)acrylamide; 1-((9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3,3-difluoro-2-methyl-1-oxopropan-2-aminium; 1-(((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3,3-difluoro-2-methyl-1-oxopropan-2-aminium; 1-((-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3,3-difluoro-2-methyl-1-oxopropan-2-aminium; 1-(((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3,3-difluoro-2-methyl-1-oxopropan-2-aminium; 3-((9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-1,1,1-trifluoro-3-oxopropan-2-aminium; 3-(((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-1,1,1-trifluoro-3-oxopropan-2-aminium; ((9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-1,1,1-trifluoro-3-oxopropan-2-aminium; (((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-1,1,1-trifluoro-3-oxopropan-2-aminium; 2-Amino-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoro-2-methylpropanamide; 2-Amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoro-2-methylpropanamide; 2-Amino-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoro-2-methylpropanamide; 2-Amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoro-2-methylpropanamide; 2-Amino-2-benzyl-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoropropanamide; 2-Amino-2-benzyl-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoropropanamide; 2-Amino-2-benzyl-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoropropanamide; 2-Amino-2-benzyl-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3,3,3-trifluoropropanamide; 2-Amino-2-cyclopropyl-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)acrylamide; 2-Amino-2-cyclopropyl-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)acrylamide; 4-(Difluoromethoxy)-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-hydroxybutyramide; (R)-4-(Difluoromethoxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-hydroxybutyramide; 2-Cyclopropyl-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide; 2-Cyclopropyl-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide; 2-Cyclopropyl-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide; 2-Cyclopropyl-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide; N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2,2-dimethylpropanamide; N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2,2-dimethylpropanamide; (S)-N-(-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxybutyramide; (S)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxybutyramide; N-((9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxybutyramide; (R)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxybutyramide; N-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide; 1-(2,3-Dihydroxypropanamido)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-7-ium hexafluorophosphate(V); (1S,9S)-1-((S)-2,3-Dihydroxypropanamido)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-7-ium hexafluorophosphate(V); 1-(2,3-Dihydroxypropanamido)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-7-ium hexafluorophosphate(V); (1S,9S)-1-((R)-2,3-Dihydroxypropanamido)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-7-ium hexafluorophosphate(V); N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-hydroxy-2-(1-(hydroxymethyl)cyclopropyl)acetamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-hydroxy-2-(1-(hydroxymethyl)cyclopropyl)acetamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methoxybutanamide; (R)-N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methoxybutanamide; (S)-N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methoxybutanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2-methylpropanamide; (S)-N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2-methylpropanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2-methylpropanamide; (R)-N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxy-2-methylpropanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-fluoro-3-hydroxypropanamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-fluoro-3-hydroxypropanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-fluoro-3-hydroxypropanamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-fluoro-3-hydroxypropanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-fluoro-3-hydroxy-2-methylpropanamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-fluoro-3-hydroxy-2-methylpropanamide; (1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-1-((2-hydroxypropyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-fluoro-3-hydroxy-2-methylpropanamide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-fluoro-3-hydroxy-2-methylpropanamide; N-(9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2,2-dimethylbutyramide; N-((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2,2-dimethylbutanamide; 2-(Difluoromethyl)-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methylbutanamide; 2-(Difluoromethyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methylbutanamide; 2-(Difluoromethyl)-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methylbutanamide; 2-(Difluoromethyl)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-4-hydroxy-2-methylbutanamide; 2-Amino-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-methylpropanamide; 2-Amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2-methylpropanamide; 1-((9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-4-methyl-1-oxopentan-2-aminium; 1 - (((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-4-methyl-1-oxopentan-2-aminium; 3 - ((9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-2-hydroxy-3-oxopropan-1-aminium; (R)-3 - (((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-2-hydroxy-3-oxopropan-1-aminium; 3 - ((9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-2-hydroxy-3-oxopropan-1-aminium; (S)-3 - (((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-2-hydroxy-3-oxopropan-1-aminium; 2-amino-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide; (S)-2-amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-3-hydroxypropanamide; 1 - ((9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3-hydroxy-1-oxopropan-2-aminium; (R)-1-(((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3-hydroxy-1-oxopropan-2-aminium; 1-((1-((9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3,3-difluoro-2-methyl-1-oxopropan-2-yl)amino)-1-oxopropan-2-aminium; (2R)-1-((1-(((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3,3-difluoro-2-methyl-1-oxopropan-2-yl)amino)-1-oxopropan-2-aminium; (2S)-1-((1-(((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3,3-difluoro-2-methyl-1-oxopropan-2-yl)amino)-1-oxopropan-2-aminium; 1-((9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3-methoxy-2-methyl-1-oxopropan-2-aminium; 1-(((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3-methoxy-2-methyl-1-oxopropan-2-aminium; 1-(((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-3-methoxy-2-methyl-1-oxopropan-2-aminium; 4-Amino-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,2-difluorobutyramide; 4-Amino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-2,2-difluorobutyramide; 2,3-Diamino-N-(9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-propanamide; (S)-2,3-Diamino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-propanamide; (R)-2,3-Diamino-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)-propanamide; (1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-1-((2-hydroxyethyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-10,13-dione; (1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-1-((3-hydroxypropyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-10,13-dione; (1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-1-((1-hydroxypropan-2-yl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-10,13-dione; (1S,9S)-1-((3,3-difluoro-2-hydroxypropyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione; 2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinolin-1-yl)amino)-N-isopropylethanamide; (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-1-((2-(hydroxymethyl)benzyl)amino)-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione; (1S,9S)-1-((2,2-difluoropropyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione; (1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1-((2,2,3,3-tetrafluoropropyl)amino)-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione; (1S,9S)-1-((1,1-difluoropropan-2-yl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione; or (1S,9S)-1-((2,2-difluoroethyl)(methyl)amino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indazino[1,2-b]quinoline-10,13-dione.

18. The compound according to claim 17 or a pharmaceutically acceptable salt or solvate thereof, which is:

19. A pharmaceutical composition comprising the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

20. Use of the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt or solvate thereof, or the pharmaceutical composition according to claim 19, in the preparation of a medicament for the treatment or prevention of cancer or tumor.

21. A method for treating or preventing cancer in a subject in need thereof, said cancer being selected from breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, gastric cancer, endometrial cancer, salivary gland cancer, esophageal cancer, melanoma, glioma, neuroblastoma, sarcoma, lung cancer (e.g., small cell lung cancer and non-small cell lung cancer), colon cancer, rectal cancer, colorectal cancer, leukemia (e.g., acute lymphoblastic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia), bone cancer, skin cancer, thyroid cancer, pancreatic cancer, and lymphoma (e.g., Hodgkin lymphoma, non-Hodgkin lymphoma, or anaplastic large cell lymphoma), said method comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound or a pharmaceutically acceptable salt or solvate thereof according to claims 1 to 18, or a pharmaceutical composition comprising said compound, salt, or solvate.

22. A method for treating and / or preventing a tumor, which comprises administering to a patient in need thereof a therapeutically effective amount of a compound or a pharmaceutically acceptable salt or solvate thereof according to claims 1 to 18, or a pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt or solvate thereof according to claims 1 to 18.

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