ATR inhibitors

Abstract

The present disclosure provides a compound of formula (I): The compound of TIFF2025512543000157.tif61160 (wherein R 1 ~R 7 and L are defined herein), or a pharma- ceutically acceptable salt thereof. Also provided are pharmaceutical compositions comprising one or more compounds described herein, and methods of treating cancer in a patient, comprising administering to the patient a compound or pharmaceutical composition described herein.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Application No. 63 / 330,944, filed April 14, 2022, the disclosure of which is incorporated herein by reference in its entirety.

[0002] (Technical field) The present disclosure relates to ATR inhibitors and methods of use thereof. [Background technology]

[0003] The ataxia-telangiectasia- and Rad3-related (ATR) protein kinase is essential for the replication stress response. ATR is a member of the phosphatidylinositol 3-kinase-related kinase (PIKK) family of kinases involved in DNA damage signaling and repair. While other members of this family (ataxia-telangiectasia-mutated (ATM) and DNA-dependent protein kinase catalytic subunit (DNA-PKcs)) are required for repair of double-strand breaks (DSBs), ATR is activated in response to single-stranded DNA (ssDNA) generated at stalled replication forks or as an intermediate in DSB repair. When replication fork progression stalls, activated ATR phosphorylates the downstream kinase Chk1, stabilizing the fork and triggering cell cycle arrest, allowing time for stress to resolve and replication to continue. Disruption of the ATR-Chk1 pathway results in stalled replication forks becoming DSBs, which, if not resolved, can cause genetic instability and adversely affect cell survival during replication. Given its crucial role in replication, ATR deficiency is lethal in early mouse embryos. However, it is important to note that the profound (>90%) suppression of ATR activity by ATR mutations is well tolerated in tissues most responsive to conventional chemotherapy, such as the bone marrow and intestinal epithelium.

[0004] ATR inhibition is synthetic lethal in cancers harboring mutations that cause oncogenic stress or aberrant DNA damage response (DDR). Cancer-associated genetic mutations activate the replication stress response and other DNA damage response (DDR) pathways. Oncogenic stresses that induce such mutations include K-Ras. G12D Mutations and H-Ras G12V These include mutations and c-Myc amplification. It has been proposed that activation of the DDR by oncogenic stress contributes to the selection of p53 and ATM mutations and loss. Mutations in the tumor suppressor p53 are found in ~50% of all human cancers. While the oncogene Myc has a similar mutation frequency, mutations in Ras family genes account for ~16% of most cancers, and the DDR protein ATM to a lesser extent. Mutations in these genes lead to increased reliance on the ATR-Chk1 pathway for genome maintenance. Studies have shown that ATR inhibition causes synthetic lethality in each of the above cancer-related conditions.

[0005] Cancers with defects in parts of the homologous recombination pathway (e.g., those containing mutations in BRCA1 and BRCA2) are highly susceptible to PARP inhibition. PARP is required for the repair of single-strand breaks (SSBs), preventing them from collapsing into DSBs, while ATR stabilizes replication forks, similarly preventing their collapse and the formation of DSBs. Thus, loss of PARP and ATR activity in either case forces cells to rely on the DSB repair pathway. Because BRCA-mutated cells are unable to repair DSBs that are susceptible to PARP inhibition, it is reasonable to assume that cells with defective DDRs (e.g., those containing BRCA mutations) will also be susceptible to ATR inhibition.

[0006] There is a need for compounds that inhibit ATR. Summary of the Invention

[0007] In some embodiments, the present disclosure provides a compound of formula (I): [ka] (In the formula, R1 ~R 7 and L is defined herein), or a pharmaceutically acceptable salt thereof.

[0008] In other embodiments, the present disclosure provides pharmaceutical compositions comprising one or more compounds described herein.

[0009] In yet other embodiments, the present disclosure provides methods of treating cancer in a patient, comprising administering to the patient a compound or pharmaceutical composition described herein. DETAILED DESCRIPTION OF THE INVENTION

[0010] The present disclosure may be more fully understood by reference to the following description, including the explanation of terms and the final examples. It is understood that some features of the compositions and methods, which for clarity are described in separate embodiments, may also be provided in combination in a single embodiment.

[0011] In the following description of examples of the present invention, all documents cited in the specification of this application, including publications, patent applications, and patents, are hereby incorporated by reference to the same extent as if each was specifically indicated by reference.

[0012] Conversely, various features of the compositions and methods, which are, for brevity, described in the context of a single embodiment, may be provided separately or in any combination.

[0013] Those skilled in the art will understand that changes may be made to the above-described examples without departing from the broad inventive concept of the present application. Accordingly, it is understood that the present invention is not limited to the examples shown and described, but that modifications are intended to be included within the spirit and scope of the present invention as defined by the appended claims. For example, certain features of the examples may or may not be a part of the claimed invention, and features of disclosed embodiments may be combined. Unless otherwise specified, the terms "a," "an," and "the" should be understood to mean "at least one" rather than being limited to one element.

[0014] The range of carbon atoms used herein (e.g., C 1-6 ) is described, all ranges and individual carbon atom numbers are included. For example, "C 1-3 "C 1-3 , C 1-2 , C 2-3 , C1, C2, and C3.

[0015] The term "alkyl" refers to alkyl groups containing 1 to 12 carbon atoms ("C 1-12 "), preferably 1 to 6 carbon atoms ("C 1-6 "). Examples of alkyl groups include methyl (Me, C1 alkyl), ethyl (Et, C2 alkyl), n-propyl (C3 alkyl), isopropyl (C3 alkyl), butyl (C4 alkyl), isobutyl (C4 alkyl), sec-butyl (C4 alkyl), tert-butyl (C4 alkyl), pentyl (C5 alkyl), isopentyl (C5 alkyl), tert-pentyl (C5 alkyl), hexyl (C6 alkyl), isohexyl (C6 alkyl), and groups that would be considered equivalent to any one of the foregoing examples in light of the ordinary skill in the art and the teachings herein. Unless otherwise specified, an alkyl group may contain one or more of OH, halogen, CN, NO2, C 1-6 Alkoxy, C 3-8 It may be optionally substituted with cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

[0016] "Cycloalkyl" refers to a saturated monocyclic or polycyclic group containing carbon and hydrogen. In some embodiments, cycloalkyl includes groups having 3 to 12 ring atoms (i.e., (C 3-12 ) cycloalkyl). Specific examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl. Unless otherwise specified, cycloalkyl groups may contain one or more OH, halogen, CN, NO, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 It may be optionally substituted with cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

[0017] "Heterocycloalkyl" refers to a saturated monocyclic or polycyclic group containing carbon and hydrogen. In some embodiments, heterocycloalkyl includes groups having 3 to 12 ring atoms and one nitrogen or oxygen atom (i.e., (C 3-12 ) heterocycloalkyl). Illustrative examples of heterocycloalkyl groups include, but are not limited to, azepanyl, azetidinyl, aziridinyl, azocanyl, azolidinyl, dioxanyl, oxetanyl, oxanyl, oxepanyl, oxiranyl, oxocanyl, oxolanyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydrofuranyl, thianyl, thiepanyl, thietanyl, thiiranyl, thiocanyl, thiolanyl, and the like. In some embodiments, heterocycloalkyl is pyrrolidinyl, tetrahydrofuranyl, oxetanyl, pyranyl, piperidinyl, or azetidinyl. Unless otherwise specified, heterocycloalkyl may be selected from the group consisting of one or more of OH, halogen, CN, NO, C, and the like. 1-6 Alkyl, C 1-6 Alkoxy, C 3-8 It may be optionally substituted with cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

[0018] "Halo" refers to fluoro, chloro, bromo, or iodo. In some embodiments, "halo" is fluoro. In other embodiments, "halo" is chloro. In still other embodiments, "halo" is bromo. In still yet other embodiments, "halo" is iodo.

[0019] "Alkoxy" refers to an alkyl group having 1 to 12 carbon atoms ("C 1-12 "), preferably 1 to 6 carbon atoms ("C 1-6 "), and straight or branched chain alkyl groups having one oxygen atom. Examples of alkyl groups include methyl (OMe), ethyl (OEt), n-propyl (O n Pr), isopropyl (O i Pr), butyl (OBu), isobutyl (O i Bu), sec-butyl (O s Bu), tert-butyl (O t Bu), pentyl (O-pentyl), isopentyl (O- i pentyl), tert-pentyl (O- t Pentyl), hexyl (O-hexyl), isohexyl (O- i hexyl), and groups that would be considered equivalent to any one of the foregoing examples in light of ordinary skill in the art and the teachings of this specification. Unless otherwise specified, alkoxy includes one or more of OH, halogen, CN, NO, C 1-6 Alkyl, C 3-8 It may be optionally substituted with cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

[0020] "Aryl" means an unsaturated ring (C 6-10 In some embodiments, aryl contains 6 to 10 ring atoms. In other embodiments, aryl contains 6 to 8 ring atoms. Aryl also includes monocyclic or fused polycyclic rings. Examples of aryl include, but are not limited to, phenyl, naphthyl, or indolyl. Unless otherwise specified, aryl includes one or more of OH, halogen, CN, NO, C. 1-6 Alkyl, C 3-8It may be optionally substituted with cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

[0021] "Heteroaryl" refers to a 5-18 membered aromatic group containing one or more ring heteroatoms (nitrogen, oxygen and / or sulfur) and which is a monocyclic, bicyclic, tricyclic or tetracyclic ring system (e.g., (C 5-13) heteroaryl). In some embodiments, heteroaryl contains 5 to 12 ring atoms. In yet other embodiments, heteroaryl contains 5 to 10 ring atoms. In other embodiments, heteroaryl contains 5 to 8 ring atoms. In yet other embodiments, heteroaryl contains 5 to 6 ring atoms. Polycyclic heteroaryls can be fused or unfused. Heteroatoms in heteroaryls can be optionally oxidized, and nitrogen atoms, if present, can be optionally quaternized. Heteroaryls can be attached to the rest of the molecule through any atom of the ring. Examples of heteroaryl include, but are not limited to, benzimidazolyl, benzindolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzoxazolyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzofurazanyl, benzothiazolyl, benzothienyl, benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furazanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, and phenyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, oxoazepinyl, oxazolyl, oxiranyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyridopyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, thiapyranyl, triazolyl, tetrazolyl, triazinyl, or thiophenyl (i.e., thienyl). Unless otherwise specified, heteroaryl is a heteroaryl group consisting of one or more of OH, halogen, CN, NO, C1-6 Alkyl, C 3-8 It may be optionally substituted with cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

[0022] "Pharmaceutically acceptable" means approved or expected to be approved by a federal or state regulatory agency or a corresponding regulatory agency in a country other than the United States, or listed in the United States Pharmacopeia or other generally recognized pharmacopeia for use in animals, especially humans.

[0023] As used herein, "subject" refers to a mammal. In some embodiments, the patient or subject is a human. In other embodiments, the patient or subject is a livestock or farm animal, a pet, or an animal commonly used for clinical experiments. In yet other embodiments, the subject is a dog, cat, or primate. The terms "human," "patient," and "subject" are used interchangeably herein.

[0024] "Treatment" of any disease or disorder, in certain embodiments, refers to ameliorating the disease or disorder (i.e., preventing or reducing the progression of the disease or at least one clinical symptom thereof). In another embodiment, "treatment" refers to improving at least one physical parameter that may not be discernible by the subject. In yet another embodiment, "treatment" refers to modulating the disease or disorder physically (e.g., stabilizing discernible symptoms), physiologically (e.g., stabilizing physical parameters), or both. In yet another embodiment, "treatment" refers to delaying the onset of the disease or disorder.

[0025] The compounds described herein may contain one or more chiral centers. Therefore, the compounds described herein may refer to a particular enantiomer or diastereomer. The compounds described herein may also be provided as a mixture of enantiomers or diastereomers.

[0026] The compounds of the present disclosure may contain any isotope of any atom contained in the compound. For example, one or more hydrogen atoms may be replaced with deuterium or tritium. Methods for synthesizing isotopes are generally known in the art.

[0027] compound The present application provides a compound of formula (I): [ka] The present invention provides a compound of the formula:

[0028] In this disclosure, R 1 is H, C 1-6 Alkyl or substituted C 1-6 In some embodiments, R 1 is H. In other embodiments, R 1 is C 1-6 alkyl. For example, R 1 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. 1 is methyl. In other embodiments, R 1 is ethyl. In yet another embodiment, R 1 is propyl. In yet another embodiment, R 1 is butyl. In yet another embodiment, R 1 is pentyl. In other embodiments, R 1 is hexyl. R 1 is optionally substituted, i.e., R 1 is a substitution C 1-6 In some embodiments, R 1 is a substitution C 1-6 In another embodiment, R 1 is substituted methyl. In yet another embodiment, R 1 is substituted ethyl. In still other embodiments, R 1 is substituted propyl. In yet another embodiment, R 1 is substituted butyl. In other embodiments, R 1is substituted pentyl. In yet another embodiment, R 1 is a substituted hexyl.

[0029] In this disclosure, R 2 is H, C 1-6 Alkyl or substituted C 1-6 In some embodiments, R 2 is H. In other embodiments, R 2 is C 1-6 alkyl. For example, R 2 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. 2 is methyl. In other embodiments, R 2 is ethyl. In yet another embodiment, R 2 is propyl. In yet another embodiment, R 2 is butyl. In yet another embodiment, R 2 is pentyl. In other embodiments, R 2 is hexyl. R 2 is optionally substituted, i.e., R 2 is a substitution C 1-6 In some embodiments, R 2 is a substitution C 1-6 In another embodiment, R 2 is substituted methyl. In yet another embodiment, R 2 is substituted ethyl. In still other embodiments, R 2 is substituted propyl. In yet another embodiment, R 2 is substituted butyl. In other embodiments, R 2 is substituted pentyl. In yet another embodiment, R 2 is a substituted hexyl.

[0030] Alternatively, R 1 and R 2 is a unitary and appropriately substituted C 3-6In some embodiments, R 1 and R 2 is unsubstituted C 3-6 Forms a cycloalkyl. For example, R 1 and R 2 are taken together to form an unsubstituted cyclopropyl. In some embodiments, R 1 and R 2 are taken together to form an unsubstituted cyclobutyl. 1 and R 2 are taken together to form an unsubstituted cyclopentyl. 1 and R 2 are taken together to form an unsubstituted cyclohexyl. In another embodiment, R 1 and R 2 is a replacement C 3-6 Forms a cycloalkyl. For example, R 1 and R 2 taken together form a substituted cyclopropyl, substituted cyclobutyl, substituted cyclopentyl, or substituted cyclohexyl. 1 and R 2 are taken together to form a substituted cyclopropyl. 1 and R 2 are taken together to form a substituted cyclobutyl. In a further embodiment, R 1 and R 2 are taken together to form a substituted cyclopentyl. 1 and R 2 are taken together to form a substituted cyclohexyl. 1 and R 2 together form an unsubstituted heterocycloalkyl. For example, R 1 and R 2 taken together form an unsubstituted pyrrolidinyl, an unsubstituted tetrahydrofuranyl, an unsubstituted oxetanyl, an unsubstituted pyranyl, an unsubstituted piperidinyl, or an unsubstituted azetidinyl. 1and R 2 are taken together to form an unsubstituted pyrrolidinyl. In other embodiments, R 1 and R 2 are taken together to form an unsubstituted tetrahydrofuranyl. 1 and R 2 are taken together to form an unsubstituted oxetanyl. 1 and R 2 are taken together to form an unsubstituted pyranyl. 1 and R 2 are taken together to form an unsubstituted piperidinyl. In other embodiments, R 1 and R 2 and together form an unsubstituted azetidinyl. In still yet another embodiment, R 1 and R 2 taken together form a substituted heterocycloalkyl. For example, R 1 and R 2 taken together form a substituted pyrrolidinyl, substituted tetrahydrofuranyl, substituted oxetanyl, substituted pyranyl, substituted piperidinyl, or substituted azetidinyl. 1 and R 2 are taken together to form a substituted pyrrolidinyl. In other embodiments, R 1 and R 2 are taken together to form a substituted tetrahydrofuranyl. 1 and R 2 are taken together to form a substituted oxetanyl. 1 and R 2 are taken together to form a substituted pyranyl. 1 and R 2 are taken together to form a substituted piperidinyl. 1 and R 2 together form a substituted azetidinyl.

[0031] In this disclosure, R 3 is H, C1-6 Alkyl or substituted C 1-6 In some embodiments, R 3 is H. In other embodiments, R 3 is C 1-6 alkyl. For example, R 3 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. 3 is methyl. In other embodiments, R 3 is ethyl. In yet another embodiment, R 3 is propyl. In yet another embodiment, R 3 is butyl. In yet another embodiment, R 3 is pentyl. In other embodiments, R 3 is hexyl. R 3 is optionally substituted, i.e., R 3 is a substitution C 1-6 In some embodiments, R 3 is a substitution C 1-6 In another embodiment, R 3 is substituted methyl. In yet another embodiment, R 3 is substituted ethyl. In still other embodiments, R 3 is substituted propyl. In yet another embodiment, R 3 is substituted butyl. In other embodiments, R 3 is substituted pentyl. In yet another embodiment, R 3 is a substituted hexyl.

[0032] In this disclosure, R 4 is H, C 1-6 Alkyl or substituted C 1-6 In some embodiments, R 4 is H. In other embodiments, R 4 is C 1-6 alkyl. For example, R 4 is methyl, ethyl, propyl, butyl, pentyl, or hexyl.4 is methyl. In other embodiments, R 4 is ethyl. In yet another embodiment, R 4 is propyl. In yet another embodiment, R 4 is butyl. In yet another embodiment, R 4 is pentyl. In other embodiments, R 4 is hexyl. R 4 is optionally substituted, i.e., R 4 is a substitution C 1-6 In some embodiments, R 4 is a substitution C 1-6 In another embodiment, R 4 is substituted methyl. In yet another embodiment, R 4 is substituted ethyl. In still other embodiments, R 4 is substituted propyl. In yet another embodiment, R 4 is substituted butyl. In other embodiments, R 4 is substituted pentyl. In yet another embodiment, R 4 is substituted hexyl. In some embodiments, R 4 is NH2, NH(C 1-6 alkyl), or NH(C 1-6 Alkyl)(C 1-6 In other embodiments, R 4 is substituted with NH. In a further embodiment, R 4 is NH(C 1-6 In still other embodiments, R 4 is NH(C 1-6 Alkyl)(C 1-6 alkyl). For example, R 4 is substituted with NHCH, NHCHCH, N(CH), or N(CHCH). 4 is substituted with NHCH. In a further embodiment, R 4 is substituted with NHCH2CH3. In yet other embodiments, R 4is substituted with N(CH). In yet another embodiment, R 4 is replaced by N(CH2CH3)2.

[0033] In this disclosure, R 5 is H, C 1-6 Alkyl or substituted C 1-6 In some embodiments, R 5 is H. In other embodiments, R 5 is C 1-6 alkyl. For example, R 5 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. 5 is methyl. In other embodiments, R 5 is ethyl. In yet another embodiment, R 5 is propyl. In yet another embodiment, R 5 is butyl. In yet another embodiment, R 5 is pentyl. In other embodiments, R 5 is hexyl. R 5 is optionally substituted, i.e., R 5 is a substitution C 1-6 In some embodiments, R 5 is a substitution C 1-6 In another embodiment, R 5 is substituted methyl. In yet another embodiment, R 5 is substituted ethyl. In still other embodiments, R 5 is substituted propyl. In yet another embodiment, R 5 is substituted butyl. In other embodiments, R 5 is substituted pentyl. In yet another embodiment, R 5 is substituted hexyl. In some embodiments, R 5 is NH2, NH(C 1-6 alkyl), or NH(C 1-6 Alkyl)(C 1-6 In other embodiments, R 5is substituted with NH. In a further embodiment, R 5 is NH(C 1-6 In still other embodiments, R 5 is NH(C 1-6 Alkyl)(C 1-6 alkyl). For example, R 5 is substituted with NHCH, NHCHCH, N(CH), or N(CHCH). 5 is substituted with NHCH. In a further embodiment, R 5 is substituted with NHCH2CH3. In yet other embodiments, R 5 is substituted with N(CH). In yet another embodiment, R 5 is replaced by N(CH2CH3)2.

[0034] In this disclosure, R 6 is H, C 1-6 Alkyl or substituted C 1-6 In some embodiments, R 6 is H. In other embodiments, R 6 is C 1-6 alkyl. For example, R 6 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. 6 is methyl. In other embodiments, R 6 is ethyl. In yet another embodiment, R 6 is propyl. In yet another embodiment, R 6 is butyl. In yet another embodiment, R 6 is pentyl. In other embodiments, R 6 is hexyl. R 6 is optionally substituted, i.e., R 6 is a substitution C 1-6 In some embodiments, R 6 is a substitution C 1-6 In another embodiment, R 6is substituted methyl. In yet another embodiment, R 6 is substituted ethyl. In still other embodiments, R 6 is substituted propyl. In yet another embodiment, R 6 is substituted butyl. In other embodiments, R 6 is substituted pentyl. In yet another embodiment, R 6 is substituted hexyl. In some embodiments, R 6 is NH2, NH(C 1-6 alkyl), or NH(C 1-6 Alkyl)(C 1-6 In other embodiments, R 6 is substituted with NH. In a further embodiment, R 6 is NH(C 1-6 In still other embodiments, R 6 is NH(C 1-6 Alkyl)(C 1-6 alkyl). For example, R 6 is substituted with NHCH, NHCHCH, N(CH), or N(CHCH). 6 is substituted with NHCH. In a further embodiment, R 6 is substituted with NHCH2CH3. In yet other embodiments, R 6 is substituted with N(CH). In yet another embodiment, R 6 is replaced by N(CH2CH3)2.

[0035] In this disclosure, R 7 is H, C 1-6 Alkyl or substituted C 1-6 In some embodiments, R 7 is H. In other embodiments, R 7 is C 1-6 alkyl. For example, R 7 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. 7 is methyl. In other embodiments, R7 is ethyl. In yet another embodiment, R 7 is propyl. In yet another embodiment, R 7 is butyl. In yet another embodiment, R 7 is pentyl. In other embodiments, R 7 is hexyl. R 7 is optionally substituted, i.e., R 7 is a substitution C 1-6 In some embodiments, R 7 is a substitution C 1-6 In another embodiment, R 7 is substituted methyl. In yet another embodiment, R 7 is substituted ethyl. In still other embodiments, R 7 is substituted propyl. In yet another embodiment, R 7 is substituted butyl. In other embodiments, R 7 is substituted pentyl. In yet another embodiment, R 7 is a substituted hexyl. 7 is NH2, NH(C 1-6 alkyl), or NH(C 1-6 Alkyl)(C 1-6 In other embodiments, R 7 is substituted with NH. In a further embodiment, R 7 is NH(C 1-6 In still other embodiments, R 7 is NH(C 1-6 Alkyl)(C 1-6 alkyl). For example, R 7 is substituted with NHCH, NHCHCH, N(CH), or N(CHCH). 7 is substituted with NHCH. In a further embodiment, R 7 is substituted with NHCH2CH3. In yet other embodiments, R 7 is substituted with N(CH). In yet another embodiment, R 7is replaced by N(CH2CH3)2.

[0036] In this disclosure, L is C 1-20 alkylene, where C 1-20 One or more carbon atoms of the alkylene may each be optionally replaced with an oxygen atom. 1-20 In another embodiment, L is a substituted C 1-20 In still other embodiments, L is C 1-18 Alkylene, C 1-16 Alkylene, C 1-14 Alkylene, C 1-12 Alkylene, C 1-10 Alkylene, C 1-8 Alkylene, C 1-6 Alkylene, or C 1-4 In some embodiments, L is C 1-18 In other embodiments, L is C 1-16 In a further embodiment, L is C 1-14 In yet another embodiment, L is C 1-12 In yet another embodiment, L is C 1-10 In other embodiments, L is C 1-8 In a further embodiment, L is C 1-6 In yet another embodiment, L is C 1-4It is alkylene. In other embodiments, L contains one or more oxygen atoms. In still other embodiments, L contains two oxygen atoms, three oxygen atoms, four oxygen atoms, five oxygen atoms, or six oxygen atoms. In some embodiments, L contains one oxygen atom. In other embodiments, L contains two oxygen atoms. In further embodiments, L contains three oxygen atoms. In still other embodiments, L contains four oxygen atoms. In yet other embodiments, L contains five oxygen atoms. In other embodiments, L contains six oxygen atoms. In some embodiments, L is -(CH2)6-. In still other embodiments, L is -CHO(CH2)4-. In other embodiments, L is -CHO(CH2)2O(CH2)2-.

[0037] The present disclosure also provides pharmaceutically acceptable salts of compounds of formula (I). "Pharmaceutically acceptable salts" refers to salts of compounds of the present disclosure that are pharmaceutically acceptable and possess the desired pharmacological activity of the parent compound. In particular, such salts are non-toxic and can be addition salts of inorganic or organic acids, as well as inorganic or organic bases. Specifically, the salts include (1) acid addition salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.), or with organic acids (e.g., acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, (2) salts formed by replacing an acidic proton present in the parent compound with a metal ion (e.g., an alkali metal ion, an alkaline earth metal ion, or an aluminum ion) or by coordination with an organic base (e.g., ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, etc.). Further examples of salts include, by way of example, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, etc., and, if the compound contains a basic functional group, salts of non-toxic organic or inorganic acids (e.g., hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate, etc.). In some embodiments, the compound is a hydrochloride (HCl) salt.

[0038] In some embodiments, the compound is [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; or [ka] or a pharmaceutically acceptable salt thereof.

[0039] composition The compounds of the present disclosure can be used alone or in combination with one or more other active ingredients to formulate pharmaceutical compositions of the present disclosure. In some embodiments, the pharmaceutical composition comprises (a) an effective amount of at least one compound described in the present disclosure, and (b) a pharmaceutically acceptable excipient.

[0040] The composition or compound may be administered by any suitable route of delivery (e.g., oral, parenteral, rectal, topical, or ocular, or inhalation). In some embodiments, the compound or composition comprising the compound is administered orally.

[0041] For oral administration, the compounds of the present disclosure may be provided in tablet or capsule form, or as a solution, emulsion, or suspension. To prepare an oral composition, the compounds may be formulated to provide a dose of, for example, about 0.05 to about 100 mg / kg per day, or about 0.05 to about 35 mg / kg per day, or about 0.1 to about 10 mg / kg per day. For example, a total dose of about 5 mg to 5 g per day may be achieved by administering the compound once, twice, three times, or four times daily.

[0042] Oral tablets or capsules may contain the compounds described in the present disclosure mixed with pharmaceutically acceptable excipients such as inert diluents, disintegrating agents, binders, lubricants, sweetening agents, flavoring agents, coloring agents, and preservatives.

[0043] The compound of the present disclosure can be administered parenterally.For example, the composition can be formulated for rectal administration as suppository.For parenteral administration, including intravenous, intramuscular, intraperitoneal or subcutaneous administration, the compound of the present disclosure can be provided in sterile aqueous solution or suspension, or parenterally acceptable oil, which is adjusted to suitable pH and osmolality.Suitable aqueous solution vehicles include Ringer's solution and isotonic sodium chloride solution.

[0044] Treatment method The macrocyclic compounds of the present disclosure, compositions comprising the same and treatment methods are useful for treating many diseases, such as cancer.In some embodiments, the compounds are useful for treating central nervous system cancer, breast cancer, pancreatic cancer, lung cancer, ovarian cancer, leukemia, lymphoma, melanoma, kidney cancer, prostate cancer, colon cancer, brain tumor, and / or glioblastoma.In other embodiments, the method treats ocular melanoma, desmoplastic round cell tumor, chondrosarcoma, meningeal carcinomatosis, diffuse large B-cell lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancer, AIDS-related lymphoma, anal cancer, rectal cancer, appendix cancer, astrocytoma, or atypical teratoid rhabdoid tumor. In yet another embodiment, the method treats basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, osteosarcoma, malignant fibrous histiocytoma, brain tumor, breast cancer, prostate cancer, bronchial tumor, Burkitt's lymphoma, or spinal cord tumor. In yet another embodiment, the method treats carcinoid tumor, cancer of unknown primary origin, atypical teratoid rhabdoid tumor of the central nervous system, leptomeningeal disease, central nervous system embryonal tumor, central nervous system lymphoma, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloproliferative disorder, colon cancer, colorectal cancer, craniopharyngioma, or cutaneous T-cell lymphoma. In a further embodiment, the method treats endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, Ewing's sarcoma family tumors, extracranial germ cell tumor, extragonadal germ cell tumor, extrahepatic bile duct cancer, or eye cancer. In yet another embodiment, the method treats gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, or glioma. In yet another embodiment, the method treats hairy cell leukemia, head and neck cancer, hepatocellular carcinoma, histiocytosis, Hodgkin's lymphoma, or hypopharyngeal cancer. In yet another embodiment, the method treats Kaposi's sarcoma or kidney cancer (renal cell carcinoma). In other embodiments, the method treats Langerhans cell histiocytosis, laryngeal cancer, lip cancer, oral cavity cancer, liver cancer, lung cancer, non-Hodgkin's lymphoma, or primary central nervous system malignant lymphoma. In yet another embodiment, the method treats Waldenstrom's macroglobulinemia (lymphoplasmacytic lymphoma), malignant fibrous histiocytoma of bone, osteosarcoma,The method treats medulloblastoma, medulloepithelioma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous cell carcinoma of the cervix with occult primary origin, multiple endocrine neoplasia, oral cancer, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, or myeloproliferative disorders. In other embodiments, the method treats cancer. In yet other embodiments, the method treats nasal cavity and paranasal sinus cancer, nasopharyngeal carcinoma, or neuroblastoma. In still yet other embodiments, the method treats oropharyngeal cancer, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian cancer, ovarian germ cell tumor, epithelial ovarian cancer, or ovarian low malignant potential tumor. In yet another embodiment, the method treats pancreatic cancer, papilloma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, intermediate pineal parenchymal tumor, pineoblastoma and supratentorial primitive neuroectodermal tumor, pituitary tumor, pleuropulmonary blastoma, gestational cancer, breast cancer, or prostate cancer. In another embodiment, the method treats rectal cancer, kidney cancer, pelvic cancer, ureteral cancer, respiratory cancer associated with the NUT gene on chromosome 15, retinoblastoma, or rhabdomyosarcoma. In yet another embodiment, the method treats high-grade prostate cancer. In yet another embodiment, the method treats intermediate-grade prostate cancer. In yet another embodiment, the method treats low-grade prostate cancer. In another embodiment, the method treats castration-resistant prostate cancer. In some embodiments, the cancer is breast cancer, prostate cancer, pancreatic cancer, lung cancer, colorectal cancer, ovarian cancer, liver cancer, melanoma, kidney cancer, central nervous system cancer, brain tumor, glioblastoma, leukemia, or lymphoma.In yet other embodiments, the method treats salivary gland cancer, non-epithelial malignant tumor, Sézary syndrome, skin cancer, eye cancer, skin malignant tumor, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, cervical squamous cell carcinoma of unknown primary origin, or supratentorial primitive neuroectodermal tumor.In yet other embodiments, the method treats T-cell lymphoma, testicular cancer, pharyngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the kidney, renal pelvis, and ureter, or gestational trophoblastic tumor.In yet another embodiment, the method treats cancer of unknown primary origin, rare childhood cancer, urethral cancer, or cancer of unknown primary origin, such as uterine sarcoma.In other embodiments, the method treats vaginal cancer,In yet another embodiment, the method treats vulvar cancer, Wilms' tumor, or gynecological cancer. In yet another embodiment, the method treats Wilms' tumor or gynecological cancer. In yet another embodiment, the method treats brain cancer, breast cancer, central nervous system cancer, colorectal cancer, glioblastoma, melanoma, leukemia, liver cancer, lung cancer, lymphoma, pancreatic cancer, prostate cancer, ovarian cancer, or kidney cancer. In yet another embodiment, the method treats ocular melanoma, desmoplastic round cell tumor, chondrosarcoma, meningeal carcinomatosis, diffuse large B-cell lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancer, AIDS-related lymphoma, anal or rectal cancer, appendix cancer, astrocytoma, atypical teratoid rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, osteosarcoma, or malignant fibrous histiocytoma, brain tumor, breast cancer, prostate cancer, bronchial tumor, gallbladder cancer, or malignant fibrous histiocytoma. Kitt lymphoma, spinal cord tumor, carcinoid tumor, cancer of unknown primary site, atypical teratoid rhabdoid tumor of the central nervous system, leptomeningeal disease, central nervous system embryonal tumor, central nervous system lymphoma, chordoma, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloproliferative disorder, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, ependymoblastoma, ependymoma, esophageal cancer, Ewing's sarcoma family of tumors, extracranial germ cell tumor, extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancer, gallbladder cancer, stomach cancer, gastrointestinal carcinoma Id tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, glioma, hairy cell leukemia, head and neck cancer, hepatocellular carcinoma (liver cancer), histiocytosis, Hodgkin's lymphoma, hypopharyngeal cancer, Kaposi's sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, lip or oral cavity cancer, lung cancer, non-Hodgkin's lymphoma, primary central nervous system malignant lymphoma, Waldenström's macroglobulinemia (lymphoplasmacytic lymphoma), medulloblastoma, medulloepithelioma, melanoma, Merkel cell carcinoma, mesothelioma, metastasis of unknown primary Metastatic squamous cell carcinoma of the neck, multiple endocrine neoplasia, oral cancer, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, myeloproliferative disorders, nasal cavity or paranasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma, oral cancer, oropharyngeal cancer, osteosarcoma or malignant fibrous histiocytoma of bone, pancreatic cancer, papilloma, paranasal sinus or nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, intermediate pineal parenchymal tumor, pineoblastoma or supratentorial primitive neuroectodermal tumor, pituitary tumor,Pleuropulmonary blastoma, prostate cancer, rectal cancer, pelvic or ureteral cancer, respiratory cancer associated with the NUT gene on chromosome 15, retinoblastoma, rhabdomyosarcoma, high-grade prostate cancer, intermediate-grade prostate cancer, low-grade prostate cancer, castration-resistant prostate cancer, salivary gland cancer, non-epithelial malignant tumors, Sézary syndrome, skin cancer including cutaneous malignancies, eye cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous cell carcinoma of the neck of unknown primary site, supratentorial primitive neuroectodermal tumor, Treating T-cell lymphoma, testicular cancer, pharyngeal cancer, thymoma or thymic carcinoma, thyroid cancer, transitional cell carcinoma of the kidney, renal pelvis, or ureter, cancer of unknown primary such as carcinoma of unknown primary, rare cancers of childhood, urethral cancer, Wilms' tumor, or gynecological cancers such as breast cancer, cervical cancer, endometrial cancer, gestational trophoblastic tumor, ovarian cancer, ovarian germ cell tumor, epithelial ovarian cancer, ovarian low malignant potential tumor, gestational cancer, uterine sarcoma, vaginal cancer, or vulvar cancer.

[0045] The usefulness of the present method and composition is not limited to any particular animal species. In at least one embodiment, the subject treated with the composition as described in the present method can be a mammal or a non-mammal. In at least one embodiment, the mammalian subject can be any mammal, including but not limited to humans, non-human primates, rodents (e.g., mice, rats, or guinea pigs), domestic pets (e.g., cats or dogs), horses, cows, pigs, sheep, goats, or rabbits. In at least one embodiment, the non-mammalian subject can be any non-mammal, including but not limited to birds (e.g., ducks, geese, chickens, or turkeys). In at least one embodiment, the subject can be male or female and of any age. In at least one embodiment, the present composition and method can be used to prevent cancer.

[0046] The compounds of the present disclosure may be used in combination with other therapeutic chemotherapeutic agents (e.g., enzyme inhibitors, PARP inhibitors, tyrosine kinase inhibitors, DNA binders, mitotic inhibitors, alkylating agents, antimetabolites, antitumor antibiotics, topoisomerase inhibitors, microtubule inhibitors, angiogenesis inhibitors, signal transduction inhibitors, cell cycle inhibitors, bisphosphonates, telomerase inhibitors, biological response modifiers (e.g., antibodies, immunotherapeutics and peptidomimetics), antihormones, antiandrogens, gene suppressors, gene activators, and antivascular agents).

[0047] The compounds described herein can be prepared, for example, according to the procedures in Schemes 1-7. [ka] [ka] [ka]

[0048] [ka] [ka]

[0049] [ka] [ka]

[0050] [ka] [ka]

[0051] [ka] [ka]

[0052] [ka]

[0053] [ka]

[0054] All documents (including publications, patent applications, and patents) cited in the specification are herein incorporated by reference to the same extent as if each was specifically indicated by reference.

[0055] Example The following examples describe the preparation of representative compounds of the invention. Melting points are given uncorrected to degrees Celsius. Mass spectral data are given as mass per unit charge (m / z), and for high-resolution mass spectral data, the calculated mass of the uncharged formula M and the experimentally observed mass ([M + H] + ) are shown. Nuclear magnetic resonance data are given as chemical shifts in parts per million (ppm) downfield from the tetramethylsilane standard, along with parameters for solvent, nuclide, and field strength. Spin-spin homonuclear coupling constants are given as J values ​​(Hertz). Multiplicities are also indicated as follows: s for singlet, d for doublet, t for triplet, q for quartet, quint for quintet, or br for broad.

[0056] Example 1: Preparation of Compound A [ka]

[0057] (i) Preparation of Compound 2 [ka] To a solution of 2-hydroxy-3-nitro-benzaldehyde (14 g, 83.77 mmol, 1.0 equiv) in MeOH (500 mL) was added MeNH (45.04 g, 435.07 mmol, 30% purity, 5.19 equiv) and stirred at 20 °C for 20 min. The reaction mixture was concentrated under reduced pressure to give a residue. The resulting mixture was then dissolved in MeOH (500 mL), and NaBHCN (7.90 g, 125.66 mmol, 1.5 equiv) was added at 0 °C. The mixture was stirred at 20 °C for 12 h, then quenched with saturated aqueous NaCO (100 mL) and extracted with EtOAc (30 mL × 3). The combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure to give 2-(methylaminomethyl)-6-nitro-phenol (15 g, crude) as an orange solid. ESI [M+H]=183.1

[0058] (ii) Preparation of Compound 3 [ka] To a solution of 2-(methylaminomethyl)-6-nitrophenol (15 g, 82.34 mmol, 1.0 equiv) in THF (300 mL) was added NaCO (8.73 g, 82.34 mmol, 1.0 equiv) and HO (14.83 g, 823.38 mmol, 14.83 mL, 10.0 equiv) until pH = 8, followed by BocO (23.36 g, 107.04 mmol, 24.59 mL, 1.3 equiv) and stirring at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove THF. The resulting residue was diluted with saturated aqueous KHSO (100 mL), extracted with ethyl acetate (300 mL x 2), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 5:1) to give tert-butyl N-[(2-hydroxy-3-nitrophenyl)methyl]-N-methyl-carbamate (21 g, 74.39 mmol, 90.35% yield) as a yellow solid. ESI [M+H-tBu] = 227.1

[0059] (iii) Preparation of Compound 4 [ka] A mixture of tert-butyl N-[(2-hydroxy-3-nitrophenyl)methyl]-N-methyl-carbamate (20.9 g, 74.04 mmol, 1.0 equiv.), Pd / C (20.9 g, 10% purity) / EtOAc (600 mL) was degassed and purged with H three times, then stirred under H (15 Psi) at 20 °C for 12 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give tert-butyl N-[(3-amino-2-hydroxy-phenyl)methyl]-N-methyl-carbamate (18 g, crude) as an orange oil. ESI [M+H-tBu] = 197.3

[0060] (iv) Preparation of Compound 5 [ka] To a solution of tert-butyl N-[(3-amino-2-hydroxyphenyl)methyl]-N-methyl-carbamate (18 g, 71.34 mmol, 1.05 equiv) and 3-amino-6-bromo-pyrazine-2-carboxylic acid (14.81 g, 67.94 mmol, 1.0 equiv) in DMF (300 mL) was added EDCI (15.63 g, 81.53 mmol, 1.2 equiv) and HOBt (4.59 g, 33.97 mmol, 0.5 equiv) at -10 °C and stirred at 20 °C for 12 h. The reaction mixture was added to HO (300 mL) and extracted with ethyl acetate (300 mL × 2). The organic layer was washed with brine (200 mL × 2), dried over NaSO, filtered, and concentrated to give a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 2:1) to give tert-butyl N-[[3-[(3-amino-6-bromo-pyrazine-2-carbonyl)amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (22 g, 47.16 mmol, 69.41% yield, 96.96% purity) as a yellow solid. ESI [M+H-Boc] = 352.1 / 354.1

[0061] (v) Preparation of Compound 7 [ka] To a solution of heptane-1,7-diol (150 g, 1.13 mol, 1.0 equiv) in THF (3000 mL) and DMF (1000 mL), NaH (45.38 g, 1.13 mol, 60% purity, 1.0 equiv) was added at 0 °C and stirred at 20 °C for 1 h. BnBr (194.06 g, 1.13 mol, 134.77 mL, 1.0 equiv) was then added at 0 °C and stirred at 20 °C for 11 h. The reaction mixture was quenched with saturated aqueous NH Cl (3 L) and extracted with ethyl acetate (3 L × 3). The organic layer was washed with brine (2 L), dried over Na SO , filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate=100:1 to 2:1) to give 7-benzyloxyheptan-1-ol (401 g, 1.80 mol, 52.99% yield) as a yellow oil. 1 H NMR (400 MHz, chloroform-d) δ 7.29-7.15 (m, 5H), 4.42 (s, 2H), 3.51 (t, J = 6.6 Hz, 2H), 3.38 (t, J = 6.6 Hz, 2H), 1.86-1.69 (m, 1H), 1.59-1.22 (m, 10H).

[0062] (vi) Preparation of Compound 8 [ka] To a solution of (COCl) (85.64 g, 674.70 mmol, 59.06 mL, 1.5 equiv) in DCM (400 mL) was added DMSO (87.86 g, 1.12 mol, 87.86 mL, 2.5 equiv) under N atmosphere at −70 °C and stirred at −70 °C for 0.5 h. A solution of 7-benzyloxyheptan-1-ol (100 g, 449.80 mmol, 1.0 equiv) in DCM (1100 mL) was then added to the above mixture at −70 °C and stirred at −70 °C for 0.5 h. TEA (250.33 g, 2.47 mol, 344.33 mL, 5.5 equiv) was then added and stirred at −70 °C for 1 h under N atmosphere. The mixture was then slowly warmed to 20 °C for 10 h under N atmosphere. The reaction mixture was quenched with saturated aqueous NH4Cl (3000 mL) and extracted with dichloromethane (3000 mL x 3). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to give 7-benzyloxyheptanal (400 g, crude) as a yellow oil. 1 H NMR (400MHz, chloroform-d)δ 9.76(s, 1H), 7.39-7.24(m, 5H), 4.51(s, 2H), 3.48(t, J=6.5Hz, 2H), 2.43(t, J=7.3Hz, 2H), 1.64(sxt, J=6.9Hz, 4H), 1.47-1.31(m, 4H)

[0063] (vii) Preparation of Compound 9 [ka] To a solution of 7-benzyloxyheptanal (100 g, 453.91 mmol, 1.0 equiv) in THF (1000 mL) was added MeMgBr (3 M, 226.96 mL, 1.5 equiv) at -10 °C under N2 atmosphere and stirred at 20 °C for 12 h under N2 atmosphere. The reaction mixture was quenched with saturated aqueous NH4Cl (2000 mL) and extracted with ethyl acetate (2000 mL x 3). The organic layer was washed with brine (150 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 100:1 to 3:1) to give 8-benzyloxyoctan-2-ol (120 g, 507.72 mmol, 27.96% yield) as a yellow oil. 1 H NMR (400 MHz, chloroform-d) δ 7.32-7.15 (m, 5H), 4.43 (s, 2H), 3.77-3.64 (m, 1H), 3.39 (t, J = 6.6 Hz, 2H), 1.59-1.50 (m, 2H), 1.40-1.21 (m, 9H), 1.13-1.07 (m, 3H).

[0064] (viii) Preparation of Compound 10 [ka] To a solution of 8-benzyloxyoctan-2-ol (60 g, 253.86 mmol, 1.0 equiv) in DCM (1000 mL) were added DIEA (131.24 g, 1.02 mol, 176.87 mL, 4.0 equiv), DMSO (79.34 g, 1.02 mol, 79.34 mL, 4.0 equiv), and then SO₃·Py (161.62 g, 1.02 mol, 4.0 equiv) at 0 °C and stirred at 30 °C for 2 h. H₂O (1000 mL) was added to the reaction mixture, which was then extracted with dichloromethane (200 mL x 3). The organic layer was washed with brine (50 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate=100:1 to 3:1) to give 8-benzyloxyoctan-2-one (80 g, 341.39 mmol, 67.24% yield) as a yellow oil.1 H NMR (400MHz, chloroform-d)δ 7.41-7.26(m, 5H), 4.52(s, 2H), 3.48(t, J=6.5Hz, 2H), 2.43(t, J=7.5Hz, 2H), 2.15(s, 3H), 1.62(quind, J=7.2, 14.6Hz, 4H), 1.46-1.28(m, 4H)

[0065] (ix) Preparation of Compound 11 [ka] To a solution of NaH (16.64 g, 416.07 mmol, 60% purity, 1.5 equiv) in THF (1000 mL) was added trimethylsulfonium iodide (73.59 g, 360.60 mmol, 1.3 equiv) in DMSO (1000 mL) at 0 °C. 8-Benzyloxyoctan-2-one (65 g, 277.38 mmol, 1.0 equiv) was then added at 0 °C and stirred at 30 °C for 12 h. The reaction mixture was quenched with saturated aqueous NH4Cl (1000 mL), concentrated to remove the solvent, and extracted with ethyl acetate (1000 mL x 3). The organic layer was washed with brine (1000 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate=100:1 to 4:1) to give 2-(6-benzyloxyhexyl)-2-methyl-oxirane (51 g, 205.35 mmol, 74.03% yield) as a yellow oil. 1 H NMR (400 MHz, chloroform-d) δ 7.32-7.14 (m, 5H), 4.43 (s, 2H), 3.39 (t, J = 6.6 Hz, 2H), 2.57-2.45 (m, 2H), 1.60-1.46 (m, 3H), 1.46-1.20 (m, 10H).

[0066] (x) Preparation of Compound 12 [ka] To a solution of 2-(6-benzyloxyhexyl)-2-methyl-oxirane (10 g, 40.26 mmol, 1.2 equiv.) and 1-bromo-4-isopropylsulfonyl-benzene (8.83 g, 33.55 mmol, 1.0 equiv.) in THF (80 mL) was added LiHMDS (1 M, 50.33 mL, 1.5 equiv.) dropwise at −10° C. under a N atmosphere and stirred at 20° C. for 12 h. The reaction mixture was quenched with saturated aqueous NH4Cl (300 mL), concentrated, the solvent removed, and the resulting mixture was extracted with ethyl acetate (500 mL). The organic layer was washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 5:1) to give 10-benzyloxy-2-(4-bromophenyl)sulfonyl-2,4-dimethyl-decan-4-ol (8 g, 15.64 mmol, 67.44% yield) as a yellow oil. ESI [M+Na + ]=533.1 / 535.1

[0067] (xi) Preparation of Compound 13 [ka] To a solution of 10-benzyloxy-2-(4-bromophenyl)sulfonyl-2,4-dimethyl-decan-4-ol (20 g, 39.10 mmol, 1.0 equiv) in DCM (300 mL) were added DHP (65.78 g, 782.00 mmol, 71.50 mL, 20.0 equiv) and PPTS (982.59 mg, 3.91 mmol, 0.1 equiv), and the mixture was stirred at 30 °C for 12 h under a N atmosphere. The reaction mixture was diluted with HO (300 mL) and extracted with dichloromethane (250 mL × 2). The organic layer was dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 3:1) to give 2-[7-benzyloxy-1-[2-(4-bromophenyl)sulfonyl-2-methyl-propyl]-1-methyl-heptoxy]tetrahydropyran (20 g, 33.58 mmol, 85.88% yield) as a yellow oil. ESI [M+Na + ]=617.2 / 619.2

[0068] (xii) Preparation of compound 14 [ka] A mixture of 2-[7-benzyloxy-1-[2-(4-bromophenyl)sulfonyl-2-methylpropyl]-1-methylheptoxy]tetrahydropyran (10 g, 16.79 mmol, 1.0 equiv.), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (5.12 g, 20.15 mmol, 1.2 equiv.), KOAc (4.94 g, 50.37 mmol, 3.0 equiv.), and Pd(dppf)Cl (1.23 g, 1.68 mmol, 0.1 equiv.) in dioxane (200 mL) was degassed and purged with N three times, and then the mixture was stirred under N at 80 °C for 2 h. The resulting reaction solution was used directly in the next step. ESI [M+Na + ]=665.3

[0069] (xiii) Preparation of Compound 15 [ka] 2-[4-(9-benzyloxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxy-nonyl)sulfonylphenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (10.8 g, 16.80 mmol, 1.0 equiv.), tert-butyl N-[[3-[(3-amino-6-bromo-pyrazine-2-carbonyl)amino]-2-hydroxyphenyl] A mixture of [methyl]-N-methyl-carbamate (7.60 g, 16.80 mmol, 1.0 equiv), Na2CO3 (2.67 g, 25.21 mmol, 1.5 equiv), Pd(dppf)Cl2 (1.23 g, 1.68 mmol, 0.1 equiv) in dioxane (250 mL) and HO (50 mL) was degassed and purged with N2 three times, then the mixture was stirred under N2 atmosphere at 80 °C for 12 h. The reaction mixture was concentrated to remove the solvent, then HO (200 mL) was added and extracted with ethyl acetate (300 mL). The organic layer was washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 1:1) to give tert-butyl N-[[3-[[3-amino-6-[4-(9-benzyloxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (27.6 g, 31.08 mmol, 92.47% yield) as a yellow oil. ESI [M+Na + ]=910.4

[0070] (xvi) Preparation of Compound 16 [ka] A mixture of tert-butyl N-[[3-[[3-amino-6-[4-(9-benzyloxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (6 g, 6.76 mmol, 1.0 equiv), Pd / C (2.6 g, 10% purity) in 90 mL of EtOAc was degassed and purged with H three times, and then the mixture was stirred under an atmosphere of H (50 psi) at 50° C. for 12 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to provide a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:tetrahydrofuran = 100:1 to 1:1) to give tert-butyl N-[[3-[[3-amino-6-[4-(9-hydroxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (4.5 g, 5.64 mmol, 41.74% yield) as a yellow oil. ESI [M+Na + ]=820.4

[0071] (xv) Preparation of Compound 17 [ka] tert-Butyl N-[[3-[[3-amino-6-[4-(9-hydroxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (4.2 g, 5.26 mmol, 1.0 equiv.), 2-(tributyl-λ 5A mixture of (2-phosphoranylidene)acetonitrile (3.81 g, 15.79 mmol, 3.0 equiv) in toluene (80 mL) was degassed and purged with N2 three times, and then the resulting mixture was stirred at 80 °C under N2 atmosphere for 12 h. The reaction mixture was diluted with HO (100 mL) and extracted with ethyl acetate (100 mL x 2). The organic layer was washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 100:1 to 1:1) to give tert-butyl N-[(5-amino-22,24,24-trimethyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-15-oxa-25λ]. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]-N-methyl-carbamate (2.6 g, 3.33 mmol, 63.33% yield) was obtained as a yellow oil. ESI [M+Na + ]=802.4

[0072] (xvi) Preparation of Compound A [ka] tert-Butyl N-[(5-amino-22,24,24-trimethyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-15-oxa-25λ] in HCl / EtOAc (25 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14A solution of ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]-N-methyl-carbamate (1.9 g, 2.44 mmol, 1.0 equiv.) was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to give a residue. The resulting residue was triturated with EtOH (20 mL) to give 5-amino-22-hydroxy-22,24,24-trimethyl-13-(methylaminomethyl)-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to obtain hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (1.34 g, 2.08 mmol, 85.21% yield, 97.93% purity, HCl) as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ 10.40(s, 1H), 8.95-9.14(m, 3H), 8.46(dd, J=1.63, 7.88Hz, 1H), 8.37(d, J=8.63Hz, 2H), 7.89-8.07(m, 4H), 7.30-7.40(m, 2H), 4.20(br t, J=5.69Hz, 2H), 3.83-3.94(m, 2H), 2.64(br t, J=5.19Hz, 3H), 1.73-1.83(m, 3H), 1.54(s, 3H), 1.48(s, 3H), 1.29-1.40(m, 5H), 1.11-1.22(m, 5H), 0.98(br d, J=6.13Hz, 2H); ESI [M+H]=596.2

[0073] Example 2: Preparation of Compound B [ka] tert-Butyl N-[(5-amino-22-methyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-spiro[15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.12,6 .0 9,14 ]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-24,1'-cyclopropan]-13-yl)methyl]-N-methyl-carbamate (compound 10, 2 g, 2.57 mmol, 1.0 equivalent) was stirred at 25 ° C for 1 hour. The mixture was concentrated under reduced pressure, and the resulting residue was purified by preparative HPLC (column: Phenomenex Luna C18 250*50 mm*10 μm; mobile phase: [water (HCl)-ACN]; B%: 20% to 50%, 10 min) to give 5-amino-22-hydroxy-22-methyl-13-(methylaminomethyl)-25,25-dioxo-spiro[15-oxa-25λ]. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaene-24,1′-cyclopropan]-7-one (1 g, 1.59 mmol, 61.72% yield, HCl) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.33(s, 1H), 9.29-9.00(m, 3H), 8.43-8.24(m, 3H), 7.98(br d, J=8.5Hz, 3H), 7.44(dd, J=1.3, 7.8Hz, 1H), 7.36-7.27(m, 1H), 4.19(br t, J=5.7Hz, 2H), 3.97-3.72(m, 2H), 2.63(br t, J=5.3Hz, 3H), 2.10 -2.00(m, 1H), 1.99-1.89(m, 1H), 1.74(tt, J=6.1, 11.9Hz, 1H), 1.68-1.54(m, 3H), 1.54-1.43(m, 2H), 1.41-1.30(m, 1H),1.21(qd, J=5.6, 11.6Hz, 1H), 1.15-1.00(m, 2H), 0.99-0.64(m, 7H); ESI [M+H]=594.4

[0074] Example 3: Preparation of Compound C [ka] tert-Butyl N-[(5-amino-22,24,24-trimethyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-15-oxa-25λ] in HCl / EtOAc (2 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 A solution of ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]carbamate (65 mg, 84.86 μmol, 1.0 equivalent) was stirred at 25 °C for 0.5 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. The resulting residue was purified by preparative HPLC (column: Phenomenex Luna C18 80*40 mm*3 μm; mobile phase: [water (0.04% HCl)-ACN]; B%: 22%~40%, 7 min) to obtain 5-amino-13-(aminomethyl)-22-hydroxy-22,24,24-trimethyl-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to obtain hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (50 mg, 77.95 μmol, 91.86% yield, 96.38% purity, HCl) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.40(s, 1H), 9.13(s, 1H), 8.48-8.24(m, 6H), 8.09-7.89(m, 4H), 7.32(d, J=4.8Hz, 2H), 4.13(br d, J=5.9Hz, 2H), 3.95-3.83(m, 2H), 1.77(br d, J=14.1Hz, 3H), 1.54(s, 3H), 1.49(s, 3H), 1.45-1.29(m, 5H), 1.23(br d, J=6.3Hz, 1H), 1.16(s, 4H), 1.01(br s, 2H); ESI [M+H]=582.3

[0075] Example 4: Preparation of Compound D Compound D was prepared using the general procedures of Examples 1-3 and 5-19.

[0076] Example 5: Preparation of Compound E [ka] 5-amino-22-hydroxy-22,24,24-trimethyl-13-(methylaminomethyl)-25,25-dioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 To a solution of hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-7-one (50 mg, 79.09 μmol, 1.0 equiv., HCl) in MeOH (2 mL), DIEA (30.66 mg, 237.26 μmol, 41.33 μL, 3.0 equiv.), AcOH (4.75 mg, 79.09 μmol, 4.52 μL, 1.0 equiv.), and HCHO (12.84 mg, 158.17 μmol, 11.78 μL, 37% purity, 2.0 equiv.) were added and stirred at 25 °C for 0.5 h. NaBHCN (4.97 mg, 79.09 μmol, 1.0 equiv.) was then added and stirred at 25 °C for 0.5 h. To this mixture was added saturated aqueous Na2CO3 (3 mL) and extracted with dichloromethane (5 mL x 2). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 10% to 40%, 8 min) to obtain 5-amino-13-[(dimethylamino)methyl]-22-hydroxy-22,24,24-trimethyl-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14] to obtain hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-7-one (30.91 mg, 47.83 μmol, 60.48% yield, 100% purity, HCl) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.42-10.29(m, 2H), 9.24-9.07(m, 2H), 8.51-8.43(m, 1H), 8.40-8.35(m, 2H), 7.92(br d, J=8.4Hz, 4H), 7.51-7.40(m, 1H), 7.34(t, J=8.1Hz, 1H), 4.32(br d, J=5.1Hz, 2H), 3.90-3.81(m, 2H), 2.77(br t, J=5.9Hz, 6H), 1.84-1.73(m, 3H), 1.59(brd, J=14.1Hz, 1H), 1.53(s, 3H), 1.48(s, 3H), 1.33(br t, J=7.5Hz, 4H), 1.24-1.14(m, 5H), 1.03-0.93(m, 2H); ESI [M+H]=610.2

[0077] Example 6: Preparation of Compound F [ka] 5-amino-22-hydroxy-22-methyl-13-(methylaminomethyl)-25,25-dioxo-spiro[15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14To a solution of ]hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-24,1'-cyclopropan]-7-one (61 mg, 96.80 μmol, 1.0 equiv., HCl) in MeOH (2 mL), DIEA (37.53 mg, 290.39 μmol, 50.58 μL, 3.0 equiv.), AcOH (5.81 mg, 96.80 μmol, 5.54 μL, 1.0 equiv.), and HCHO (15.71 mg, 193.59 μmol, 14.41 μL, 37% purity, 2.0 equiv.) were added and stirred at 25 °C for 0.5 h. NaBHCN (6.08 mg, 96.80 μmol, 1.0 equiv.) was then added and stirred at 25 °C for 1 h. To this mixture was added saturated aqueous Na2CO3 (3 mL) and extracted with dichloromethane (5 mL x 2). The organic layer was dried over Na2SO4, filtered, and concentrated to obtain a residue. The resulting residue was then purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 10%~40%, 8 min) to obtain 5-amino-13-[(dimethylamino)methyl]-22-hydroxy-22-methyl-25,25-dioxo-spiro[15-oxa-25λ]. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaene-24,1′-cyclopropan]-7-one (46.45 mg, 72.10 μmol, 74.49% yield, 100% purity, HCl) was obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ 10.34(s, 1H), 10.12(br s, 1H), 9.09(s, 1H), 8.42-8.27(m, 3H), 7.98(br d, J=7.9Hz, 4H), 7.52-7.39(m, 1H), 7.34(br t, J=7.8Hz, 1H), 4.34(br s, 2H), 3.88-3.77(m, 2H), 2.79(br s, 6H), 2.09-2.01(m, 1H),1.98-1.89(m, 1H), 1.74(br d, J=8.3Hz, 1H), 1.68-1.56(m, ESI [M+H]=608.4

[0078] Example 7: Preparation of Compound H-2 [ka]

[0079] (i) Preparation of Compound 2 [ka] To a solution of oct-7-enylacetic acid (9.4 g, 55.21 mmol, 1.0 equiv) in DCM (500 mL) was added m-CPBA (33.63 g, 165.64 mmol, 85% purity, 3.0 equiv) at 0 °C and stirred at 25 °C for 12 h. The reaction mixture was quenched with saturated aqueous NaSO (500 mL) and extracted with dichloromethane (800 mL × 2). The organic layer was washed with saturated aqueous NaHCO (300 mL × 2), dried over NaSO, filtered, and concentrated to give 6-(oxiran-2-yl)hexylacetic acid (10.2 g, crude) as a yellow oil. 1H NMR (400MHz, chloroform-d) δ 3.99(t, J=6.7Hz, 2H), 2.87-2.81(m, 1H), 2.68(t, J=4.4Hz, 1H), 2.40(dd, J=2.7, 4.8Hz, 1H), 1.98(s, 3H), 1.60-1.52(m, 2H), 1.51-1.37(m, 4H), 1.31(br d, J=3.6Hz, 4H)

[0080] (ii) Preparation of Compound 3 [ka] To a solution of 6-(oxiran-2-yl)hexyl acetate (10.2 g, 54.77 mmol, 1.0 equiv) in MeOH (150 mL) was added K2CO3 (11.35 g, 82.15 mmol, 1.5 equiv) and stirred at 25 °C for 1 h. The mixture was filtered and concentrated to give a residue. The resulting residue was diluted with HO (100 mL) and extracted with dichloromethane:methanol = 8:1 (300 mL × 2). The organic layer was dried over Na2SO4, filtered, and concentrated to give 6-(oxiran-2-yl)hexan-1-ol (7.9 g, crude) as a yellow oil. 1 H NMR (400 MHz, chloroform-d) δ 3.58 (t, J = 6.5 Hz, 2H), 2.88-2.79 (m, 1H), 2.68 (t, J = 4.5 Hz, 1H), 2.40 (dd, J = 2.8, 4.8 Hz, 1H), 1.56-1.31 (m, 10H).

[0081] (iii) Preparation of Compound 4 [ka] To a solution of 6-(oxiran-2-yl)hexan-1-ol (7.9 g, 54.78 mmol, 1.0 equiv) in DCM (100 mL) was added TEA (16.63 g, 164.34 mmol, 22.87 mL, 3.0 equiv) and DMAP (669.25 mg, 5.48 mmol, 0.1 equiv). TBDPSCl (18.07 g, 65.74 mmol, 16.89 mL, 1.2 equiv) was then added at 0 °C, and the mixture was stirred at 25 °C for 12 h. The reaction mixture was diluted with HO (50 mL) and extracted with dichloromethane (100 mL x 2). The organic layer was dried over NaSO, filtered, and concentrated to give a residue. The resulting residue was then purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 100:1 to 0:1) to give tert-butyl-[6-(oxiran-2-yl)hexoxy]-diphenyl-silane (14.9 g, 38.94 mmol, yield 71.09%) as a yellow oil. 1 H NMR (400MHz, chloroform-d) δ 7.59(dd, J=1.5, 7.8Hz, 4H), 7.38-7.27(m, 6H), 3.58(t, J=6.4Hz, 2H), 2.85 -2.79(m, 1H), 2.67(t, J=4.5Hz, 1H), 2.38(dd, J=2.8, 5.0Hz, 1H), 1.52-1.26(m, 10H), 0.97(s, 9H)

[0082] (iv) Preparation of Compound 5 [ka] To a mixture of tert-butyl-[6-(oxiran-2-yl)hexoxy]-diphenyl-silane (6.08 g, 15.88 mmol, 1.1 equiv) and 1-bromo-4-isopropylsulfonylbenzene (3.8 g, 14.44 mmol, 1.0 equiv) in THF (80 mL) was added LiHMDS (1 M, 21.66 mL, 1.5 equiv) dropwise at -10 °C and stirred at 15 °C under a N atmosphere for 5 h. The mixture was quenched with saturated aqueous NH Cl (60 mL) at 0 °C and extracted with ethyl acetate (100 mL × 2). The organic layer was dried over Na SO , filtered, and concentrated to give a residue. The resulting residue was then purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 0:1) to give 2-(4-bromophenyl)sulfonyl-10-[tert-butyl(diphenyl)silyl]oxy-2-methyl-decan-4-ol (14.1 g, 21.83 mmol, 75.60% yield) as a yellow oil. 1 H NMR (400MHz, chloroform-d) δ 7.78-7.68(m, 8H), 7.48-7.38(m, 6H), 3.90(br d, J=1.2Hz, 1H), 3.68(br t, J=6.3Hz, 2H), 1.59(br d, J=2.4Hz, 2H), 1.48-1.28(m, 16H), 1.08(s, 9H)

[0083] (v) Preparation of Compound 6 [ka] To a solution of 2-(4-bromophenyl)sulfonyl-10-[tert-butyl(diphenyl)silyl]oxy-2-methyl-decan-4-ol (14.1 g, 21.83 mmol, 1.0 equiv) in DCM (200 mL) was added DHP (45.92 g, 545.87 mmol, 49.91 mL, 25.0 equiv) and PPTS (548.71 mg, 2.18 mmol, 0.1 equiv) and stirred at 40 °C for 12 h under a N atmosphere. HO (100 mL) was added to the mixture, which was then extracted with dichloromethane (200 mL × 2). The organic layer was dried over NaSO, filtered, and concentrated to give a residue. The resulting residue was then purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 0:1) to give [9-(4-bromophenyl)sulfonyl-9-methyl-7-tetrahydropyran-2-yloxy-decoxy]-tert-butyl-diphenyl-silane (13.5 g, 18.50 mmol, 84.71% yield) as a yellow oil. 1 H NMR (400MHz, chloroform-d) δ 7.77-7.66(m, 8H), 7.48-7.37(m, 6H), 4.62-4.57(m, 1H), 3.95-3.84(m, 2H),3.79-3.71(m, 1H), 3.68(dt, ESI [M+Na + ]=753.2

[0084] (vi) Preparation of Compound 7 [ka] A mixture of [9-(4-bromophenyl)sulfonyl-9-methyl-7-tetrahydropyran-2-yloxy-decoxy]-tert-butyl-diphenyl-silane (2 g, 2.74 mmol, 1.0 equiv.), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (765.42 mg, 3.01 mmol, 1.1 equiv.), Pd(dppf)Cl (200.50 mg, 274.02 μmol, 0.1 equiv.), and KOAc (537.86 mg, 5.48 mmol, 2.0 equiv.) in dioxane (30 mL) was degassed and purged with N three times, then stirred at 80 °C under N for 1 h. The reaction solution was used directly in the next step. ESI [M+Na + ]=799.4

[0085] (vii) Preparation of Compound 8 [ka] tert-Butyl-[9-methyl-7-tetrahydropyran-2-yloxy-9-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]sulfonyldecoxy]-diphenyl-silane (2.1 g, 2.70 mmol, 1.0 equiv.), tert-butyl N-[[3-[(3-amino-6-bromo-pyrazine-2-carbonyl)amino]-2-hydroxy- A mixture of [phenyl]methyl]carbamate (1.07 g, 2.43 mmol, 0.9 equiv), Na2CO3 (572.96 mg, 5.41 mmol, 2.0 equiv), and Pd(PPh3)4 (624.67 mg, 540.58 μmol, 0.2 equiv) in dioxane (30 mL) and HO (6 mL) was degassed and purged with N2 three times, and the resulting mixture was stirred at 80 °C under a N2 atmosphere for 12 h. The mixture was filtered, concentrated, diluted with HO (50 mL), and extracted with ethyl acetate (100 mL × 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated to give a residue. The resulting residue was then purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 0:1) to give tert-butyl N-[[3-[[3-amino-6-[4-[9-[tert-butyl(diphenyl)silyl]oxy-1,1-dimethyl-3-tetrahydropyran-2-yloxynonyl]sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]carbamate (2.3 g, 2.28 mmol, 84.39% yield) as a yellow solid. 1H NMR (400MHz, クロロホルム-d) δ 10.61(br s, 1H), 9.77(br s, 1H), 8.69(d, J=3.5Hz, 1H), 8.27(br s, 1H), 8.13 -8.06(m, 2H), 7.92(d, J=8.1Hz, 2H), 7.59(br d, J=7.4Hz, 4H), 7.38-7.27(m, 6H), 6.88-6.80(m, 2H), 4.57-4.39(m, 1H),4.20(d, J=6.6Hz, 2H), 3.92-3.66(m, 2H), 3.57(dt, J=3.0, 6.4Hz, 2H), 3.45-3.28(m, 1H), 2.03-1.93(m, 2H), 1.89-1.77(m, 1H), 1.75-1.54(m, 3H), 1.53-1.39(m, 14H), 1.35-1.16(m, 13H), 0.97(d, J=2.6Hz, 9H); ESI [M+H-THP-Boc]=824.4

[0086] (viii) Production of Compound 9

change

[0087] (ix) Preparation of Compound 10 [ka] To a solution of tert-butyl N-[[3-[[3-amino-6-[4-(9-hydroxy-1,1-dimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]carbamate (750 mg, 974.10 μmol, 1.0 equivalent) in toluene (80 mL) was added a solution of 2-(tributyl-5-phosphoranylidene)acetonitrile (705.30 mg, 2.92 mmol, 3.0 equivalent) in toluene (3 mL) at 0° C., and the mixture was stirred at 80° C. for 12 hours. The mixture was concentrated, and the resulting residue was purified by column chromatography (SiO2, petroleum ether:tetrahydrofuran=100:1 to 1:1) and purified with tert-butyl N-[(5-amino-24,24-dimethyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonane-13-yl)methyl]carbamate (800 mg, crude) was obtained as a yellow oil. ESI [M+H-THP-Boc] = 568.3

[0088] (x) Preparation of Compound 11 [ka] tert-Butyl N-[(5-amino-24,24-dimethyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14To a solution of [Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]carbamate (700 mg, 930.94 μmol, 1.0 equiv.) in EtOH (10 mL) was added 4-methylbenzenesulfonic acid hydrate (177.08 mg, 930.94 μmol, 1.0 equiv.) and stirred at 30 °C for 3 h. The mixture was quenched with saturated aqueous Na2CO3 (10 mL) at 0 °C and extracted with ethyl acetate (30 mL x 3). The organic layer was dried over Na2SO4, filtered, and concentrated to give a residue. The resulting residue was then purified by column chromatography (SiO2, petroleum ether:tetrahydrofuran = 100:1 to 0:1) and purified with tert-butyl N-[(5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]carbamate (440 mg, 658.87 μmol, 70.77% yield) was obtained as a yellow solid. ESI [M+H-Boc] = 568.2

[0089] (xi) Preparation of Compounds 12A and 12B [ka] The racemate was purified by SFC (column: DAICEL Chiralpak AD (250 mm * 30 mm, 10 μm); mobile phase: [0.1% NH3H2O / EtOH]; B%: 46% - 46%, 8 min) and arbitrarily assigned as follows: tert-Butyl N-[[(22R)-5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]carbamate (peak 1, retention time = 1.813 min, 70 mg, 104.82 μmol, 35.00% yield, ee% = 100%, yellow solid), and tert-Butyl N-[[(22S)-5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]carbamate (Peak 2, retention time = 2.038 min, 80 mg, 119.79 μmol, 40.00% yield, ee% = 96.02%, yellow solid) ESI [M+H-Boc]=568.2

[0090] (xii) Preparation of Compound H-2 [ka] tert-Butyl N-[[(22R)-5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ in HCl / EtOAc (2 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]carbamate (70 mg, 104.82 μmol, 1.0 equiv.) was stirred at 20 ° C. for 1 hour. The mixture was concentrated, and the resulting residue was purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 20% to 70%, 8 min) to give (22R)-5-amino-13-(aminomethyl)-22-hydroxy-24,24-dimethyl-25,25-dioxo-15-oxa-25λ. 6-Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to obtain hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (46.27 mg, 76.59 μmol, 73.06% yield, 100% purity, HCl) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.40(s, 1H), 9.13(s, 1H), 8.49-8.30(m, 6H), 8.15-7.84(m, 4H), 7.37-7.27(m, 2H), 4.12(q, J=5.5Hz, 2H), 3.94-3.88(m, 2H), 3.60-3.53(m, 1H), 1.78(quin, J=7.3Hz, 2H), 1.68(br d, J=13.9Hz, 1H), 1.52 -1.38(m, 7H), 1.34(s, 4H), 1.26-1.16(m, 2H), 1.15-1.06(m, 2H); ESI [M+H]=568.4

[0091] Example 8: Preparation of Compound H-1 [ka] tert-Butyl N-[[(22S)-5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ in HCl / EtOAc (2 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]carbamate (80 mg, 119.79 μmol, 1.0 equiv.) was stirred at 20 ° C. for 1 hour. The mixture was concentrated, and the resulting residue was purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 20% to 70%, 8 min) to give (22S)-5-amino-13-(aminomethyl)-22-hydroxy-24,24-dimethyl-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to obtain hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (42.58 mg, 70.48 μmol, 58.83% yield, 100% purity, HCl) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.40(s, 1H), 9.13(s, 1H), 8.46(dd, J=2.3, 7.3Hz, 1H), 8.41-8.28(m, 5H), 8.12-7.90(m, 4H), 7.36-7.28(m, 2H), 4.12(q, J=5.4Hz, 2H), 3.96-3.82(m, 2H), 3.55(br d, J=9.4Hz, 1H), 1.83-1.74(m, 2H), 1.68(br d, J=13.8Hz, 1H), 1.51-1.38(m, 7H), 1.38-1.31(m, 4H), 1.20(td, J=7.3, 14.5Hz, 2H), 1.16-1.07(m, 2H); ESI [M+H]=568.4

[0092] Example 9: Preparation of Compound G [ka]

[0093] (i) Preparation of Compound 2 [ka] tert-Butyl N-[(5-amino-24,24-dimethyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-15-oxa-25λ] in HCl / EtOAc (3 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 [Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]-N-methyl-carbamate (270 mg, 352.50 μmol, 1.0 equiv.) was stirred at 15° C. for 1 hour. The combined organic layer was concentrated to give 5-amino-22-hydroxy-24,24-dimethyl-13-(methylaminomethyl)-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to give hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (210 mg, crude, HCl) as a yellow solid. ESI [M+H]=582.4

[0094] (ii) Preparation of Compound G [ka] 5-amino-22-hydroxy-24,24-dimethyl-13-(methylaminomethyl)-25,25-dioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14To a solution of hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (200 mg, 323.53 μmol, 1.0 equiv., HCl) in MeOH (5 mL), DIEA (125.44 mg, 970.58 μmol, 3.0 equiv.), AcOH (19.43 mg, 323.53 μmol, 1.0 equiv.), and HCHO (52.51 mg, 647.05 μmol, 37% purity, 2.0 equiv.) were added and stirred at 25 °C for 0.5 h. NaBHCN (20.33 mg, 323.53 μmol, 1.0 equiv.) was then added and stirred at 25 °C for 1 h. Saturated aqueous NaCO solution (5 mL) was added to the mixture, which was then extracted with dichloromethane (10 mL x 2). The organic layer was dried over Na2SO4, filtered, and concentrated to obtain a residue, which was then purified by preparative HPLC (column: Phenomenex C18 80*40mm*3μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 35%~65%, 8 min) to obtain 5-amino-13-[(dimethylamino)methyl]-22-hydroxy-24,24-dimethyl-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (42.42 mg, 70.58 μmol, 21.82% yield, 99.127% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.39(s, 1H), 9.10(s, 1H), 8.41-8.30(m, 3H), 8.22-7.83(m, 4H), 7.17(d, J=5.3Hz, 2H), 4.50(d, J=5.6Hz, 1H), 3.96-3.80(m, 2H), 3.55(br d, J=4.0Hz, 1H), 3.50-3.40(m, 2H), 2.19(s, 6H), 1.80-1.65(m, 3H), 1.52-1.46(m, 4H), 1.46-1.37(m, 3H), 1.37-1.30(m, 4H), 1.25-1.07(m, 4H); ESI [M+H]=596.2

[0095] Example 10: Preparation of Compound G-2 [ka] (i) Preparation of Compound 2 [ka] A mixture of [9-(4-bromophenyl)sulfonyl-9-methyl-7-tetrahydropyran-2-yloxy-decoxy]-tert-butyl-diphenyl-silane (2 g, 2.74 mmol, 1.0 equiv.), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (765.42 mg, 3.01 mmol, 1.1 equiv.), Pd(dppf)Cl2 (200.50 mg, 274.02 μmol, 0.1 equiv.), and KOAc (537.86 mg, 5.48 mmol, 2.0 equiv.) in dioxane (30 mL) was stirred at 80 °C under a N2 atmosphere for 1 hour. The resulting reaction solution was used directly in the next step. ESI [M+Na + ]=799.4

[0096] (iii) Preparation of Compound 3 [ka] tert-Butyl N-[[3-[(3-amino-6-bromo-pyrazine-2-carbonyl)amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (1.10 g, 2.43 mmol, 0.9 equiv.), tert-butyl-[9-methyl-7-tetrahydropyran-2-yloxy-9-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 A mixture of [-yl)phenyl]sulfonyl-decoxy]-diphenyl-silane (2.1 g, 2.70 mmol, 1.0 equiv.), Na2CO3 (572.35 mg, 5.40 mmol, 2.0 equiv.), and Pd(dppf)Cl2 (395.12 mg, 540.00 μmol, 0.2 equiv.) in dioxane (30 mL) and HO (6 mL) was stirred at 80 °C under a N2 atmosphere for 12 h. The mixture was filtered and concentrated, and the resulting residue was diluted with HO (50 mL) and extracted with ethyl acetate (100 mL × 2). The organic layer was then dried over Na2SO4, filtered, and concentrated to give a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 0:1) to give tert-butyl N-[[3-[[3-amino-6-[4-[9-[tert-butyl(diphenyl)silyl]oxy-1,1-dimethyl-3-tetrahydropyran-2-yloxynonyl]sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (2.5 g, 2.45 mmol, 90.57% yield) as a yellow oil. 1H NMR (400MHz, クロロホルム-d) δ 10.73(br s, 1H), 10.30-10.04(m, 1H), 8.76(br s, 1H), 8.45(br d, J=3.7Hz, 1H), 8.18(br d, J=1.6Hz, 2H), 7.99(br d, J=2.6Hz, 2H), 7.67(br s, 4H), 7.40(br s, 6H), 6.92(br d, J=1.7Hz, 2H), 4.67-4.46(m, 1H), 4.37(br s, 2H), 3.98-3.77(m, 2H), 3.64(br s, 2H), 3.54-3.34(m, 1H), 2.92(br s, 3H), 2.08-2.01(m, 2H), 1.86(br d, J=1.7Hz, 4H), 1.81-1.63(m, 6H), 1.51(br s, 14H), 1.41-1.32(m, 7H), 1.04(br s, 9H); ESI [M+H-THP-Boc]=838.4

[0097] (iii) Production of compound 4

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[0098] (iv) Preparation of Compound 5 [ka] To a solution of tert-butyl N-[[3-[[3-amino-6-[4-(9-hydroxy-1,1-dimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (300 mg, 382.67 μmol, 1.0 equiv.) in toluene (5 mL) was added 2-(tributyl-λ)- 5 A solution of 277.07 mg (1.15 mmol, 3.0 equivalents) of N-phosphoranylidene acetonitrile in toluene (1 mL) was added at 0°C, and the mixture was stirred at 80°C for 12 hours under a N2 atmosphere. The combined organic layers were concentrated and purified by column chromatography (SiO2, petroleum ether:tetrahydrofuran = 100:1 to 0:1) to obtain tert-butyl N-[(5-amino-24,24-dimethyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-15-oxa-25λ].6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]-N-methyl-carbamate (280 mg, crude) was obtained as a yellow viscous material. ESI [M+H-THP-Boc]=582.3

[0099] (v) Preparation of Compound 6 [ka] tert-Butyl N-[(5-amino-24,24-dimethyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 To a solution of [Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]-N-methylcarbamate (1.15 g, 1.50 mmol, 1.0 equiv) in EtOH (10 mL) was added 4-methylbenzenesulfonic acid hydrate (285.59 mg, 1.50 mmol, 1.0 equiv) and stirred at 30 °C for 3 h. The mixture was diluted with ice water, adjusted to pH 7 with saturated aqueous Na2CO3 (10 mL), and extracted with ethyl acetate (50 mL x 2). The reaction mixture was then dried over Na2SO4, filtered, and concentrated to give a residue. The resulting residue was purified by column chromatography (SiO2, petroleum ether:tetrahydrofuran = 50:1 to 0:1) to obtain tert-butyl N-[(5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]-N-methyl-carbamate (630 mg, 923.97 μmol, 61.54% yield) was obtained as a yellow solid. ESI [M+H-Boc]=582.4

[0100] (vi) Preparation of Compounds 7A and 7B [ka] The racemate was purified by SFC (column: Phenomenex-Cellulose-2 (250 mm*30 mm, 10 μm); mobile phase: [MeOH-ACN]; B%: 70%-70%, 15 min) and arbitrarily assigned as follows: tert-Butyl N-[[(22R)-5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]-N-methyl-carbamate (peak 1, retention time = 2.255 min, 90 mg, 132.00 μmol, 45.00% yield, ee% = 99.64%, yellow solid) and tert-Butyl N-[[(22S)-5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]-N-methyl-carbamate (Peak 2, Retention time = 3.724 min, 110 mg, 161.33 μmol, 55.00% yield, ee% = 99%, yellow solid) ESI [M+H-Boc] = 582.4

[0101] (vii) Preparation of Compound G-2 [ka] tert-Butyl N-[[(22R)-5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 A solution of ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]-N-methyl-carbamate (80 mg, 117.33 μmol, 1.0 equiv.) in HCl / EtOAc (1 mL, 4 M) was stirred for 1 h at 20 ° C. The mixture was concentrated and purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 20% to 70%, 8 min) to give (22R)-5-amino-22-hydroxy-24,24-dimethyl-13-(methylaminomethyl)-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to obtain hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (56.83 mg, 91.93 μmol, 78.35% yield, 100% purity, HCl) as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ 10.39(s, 1H), 9.16-9.03(m, 3H), 8.50-8.45(m, 1H), 8.38(d, J=8.4Hz, 2H), 8.10-7.96(m, 1H), 7.92(d, J=8.4Hz, 2H), 7.41-7.29(m, 2H), 4.19(br t, J=5.8Hz, 2H), 3.95-3.81(m, 2H), 3.61-3.50(m, 1H), 2.63(t, J=5.3Hz, 3H), 1.85-1.72(m, 2H), 1.67(brd, J=13.8Hz, 1H), 1.52-1.30(m, 11H), 1.19(td, J=6.8, 13.7Hz, 2H), 1.15-1.06(m, 2H); ESI [M+H]=582.4

[0102] Example 11: Preparation of Compound G-1 [ka] tert-Butyl N-[[(22S)-5-amino-22-hydroxy-24,24-dimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 A solution of ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]-N-methyl-carbamate (100 mg, 146.66 μmol, 1.0 equiv.) in HCl / EtOAc (1 mL, 4 M) was stirred for 1 h at 20 ° C. The mixture was concentrated and purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water(HCl)-ACN]; B%: 20%~70%, 8 min) to give (22S)-5-amino-22-hydroxy-24,24-dimethyl-13-(methylaminomethyl)-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14]ヘントリアコンタ-1(28),2(31),3,5,9(14),10,12,26,29-ノナエン- 7-オン (53.37 mg, 91.74 μmol, yield 62.56%, purity 100%) was obtained as a yellow solid. 1 H NMR(400MHz, DMSO-d6) δ 10.39(s, 1H), 9.14(s, 1H), 9.08(br d, J=3.4Hz, 2H), 8.48(dd, J=1.4, 8.1Hz, 1H), 8.38(d, J=8.5Hz, 2H), 8.10-7.96(m, 1H), 7.92(d, J=8.5Hz, 2H), 7.48-7.25(m, 2H), 4.19(br t, J=5.8Hz, 2H), 3.96-3.79(m, 2H), 3.61-3.50(m, 1H), 2.63(t, J=5.3Hz, 3H), 1.88-1.73(m, 2H), 1.67(br d, J=13.8Hz, 1H), 1.51-1.30(m, 11H), 1.20(td, J=7.0, 13.9Hz, 2H), 1.14-1.03(m, 2H); ESI [M+H]=582.4

[0103] Example 12: Production of Compound C-2

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[0104] (i) Production of Compound 2

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[0105] (ii) Preparation of Compound 3 [ka] A mixture of tert-butyl N-[(2-hydroxy-3-nitrophenyl)methyl]carbamate (2.4 g, 8.95 mmol, 1.0 equiv.), Pd / C (10% purity) / EtOAc (160 mL) was degassed and purged with H three times, and the resulting mixture was stirred under H (15 psi) at 30 °C for 12 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give tert-butyl N-[(3-amino-2-hydroxyphenyl)methyl]carbamate (2.2 g, crude) as a yellow oil. ESI [M+H-tBu] = 183.1

[0106] (iii) Preparation of Compound 4 [ka] To a solution of 3-amino-6-bromo-pyrazine-2-carboxylic acid (1.8 g, 8.26 mmol, 1.0 equiv) and tert-butyl N-[(3-amino-2-hydroxyphenyl)methyl]carbamate (2.16 g, 9.08 mmol, 1.1 equiv) in DMF (40 mL) was added EDCI (1.90 g, 9.91 mmol, 1.2 equiv) and HOBt (557.81 mg, 4.13 mmol, 0.5 equiv) at -10 °C and stirred at 20 °C for 12 h. The reaction mixture was diluted with HO (50 mL) and extracted with ethyl acetate (50 mL × 2). The organic layer was washed with brine (20 mL × 3), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:tetrahydrofuran = 100:1 to 0:1) and then triturated with EtOH (20 mL) to give tert-butyl N-[[3-[(3-amino-6-bromo-pyrazine-2-carbonyl)amino]-2-hydroxy-phenyl]methyl]carbamate (1.5 g, 3.42 mmol, 41.45% yield) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.18(s, 1H), 9.62(br s, 1H), 8.47(s, 1H), 8.08(br d, J=7.0Hz, 1H), 7.82(br s, 2H), 7.53(br t, J=5.1Hz, 1H), 6.91(q, J=7.8Hz, 2H), 4.16(br d, J=6.1Hz, 2H), 1.43(s, 9H); ESI [M+H-tBu]=382.1 / 384.1

[0107] (iv) Preparation of Compound 6 [ka] A mixture of 2-[7-benzyloxy-1-[2-(4-bromophenyl)sulfonyl-2-methylpropyl]-1-methylheptoxy]tetrahydropyran (1.25 g, 2.10 mmol, 1.0 equiv), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (639.51 mg, 2.52 mmol, 1.2 equiv), KOAc (617.89 mg, 6.30 mmol, 3.0 equiv), and Pd(dppf)Cl (153.56 mg, 209.86 μmol, 0.1 equiv) in dioxane (20 mL) was degassed and purged with N three times, and the resulting mixture was stirred at 80 °C under N for 2 h. This reaction solution was used directly in the next step. + ]=665.3

[0108] (v) Preparation of Compound 7 [ka] To a solution of 2-[4-(9-benzyloxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxy-nonyl)sulfonylphenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.35 g, 2.10 mmol, 1.0 equiv.) and tert-butyl N-[[3-[(3-amino-6-bromo-pyrazine-2-carbonyl)amino]-2-hydroxyphenyl]methyl]carbamate (920.61 mg, 2.10 mmol, 1.0 equiv.) in dioxane (25 mL) and HO (5 mL) was added Pd(dppf)Cl (153.70 mg, 210.05 μmol, 0.1 equiv.) and NaCO (333.95 mg, 3.15 mmol, 1.5 equiv.) and stirred at 80 °C for 12 h. The reaction mixture was concentrated to remove the solvent, then HO (50 mL) was added and extracted with ethyl acetate (50 mL). The organic layer was washed with brine (50 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:ethyl acetate = 50:1 to 1:1) to give tert-butyl N-[[3-[[3-amino-6-[4-(9-benzyloxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxyphenyl]methyl]carbamate (1.45 g, crude) as a yellow solid. ESI [M+H + ]=874.3

[0109] (vi) Preparation of Compound 8 [ka] A mixture of tert-butyl N-[[3-[[3-amino-6-[4-(9-benzyloxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]carbamate (50 mg, 57.20 μmol, 1.0 equiv), Pd / C (50 mg, 10% purity) in 2 mL of EtOAc was degassed and purged with H three times, and the resulting mixture was stirred under an atmosphere of H (30 psi) at 35° C. for 5 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to provide a residue. The resulting residue was purified by column chromatography (SiO, petroleum ether:tetrahydrofuran = 50:1 to 1:1) to give tert-butyl N-[[3-[[3-amino-6-[4-(9-hydroxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxyphenyl]methyl]carbamate (45 mg, crude) as a yellow solid. ESI [M+ Na + ]=806.4

[0110] (vii) Preparation of Compound 9 [ka] tert-Butyl N-[[3-[[3-amino-6-[4-(9-hydroxy-1,1,3-trimethyl-3-tetrahydropyran-2-yloxynonyl)sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]carbamate (70 mg, 89.29 μmol, 1.0 equiv.), 2-(tributyl-λ 5A mixture of (-phosphoranylidene)acetonitrile (43.10 mg, 178.58 μmol, 2.0 equiv) in toluene (2 mL) was degassed and purged with N three times, and the resulting mixture was stirred at 80 °C under N atmosphere for 12 h. The reaction mixture was diluted with HO (20 mL) and extracted with ethyl acetate (30 mL × 2). The organic layer was dried over NaSO, filtered, concentrated under reduced pressure, and purified by tert-butyl N-[(5-amino-22,24,24-trimethyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-15-oxa-25λ]. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]carbamate (65 mg, crude) was obtained as a yellow solid. ESI [M+ Na + ]=788.4

[0111] (viii) Preparation of Compound 11 [ka] 5-amino-13-(aminomethyl)-22-hydroxy-22,24,24-trimethyl-25,25-dioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14To a solution of hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (80 mg, 129.41 μmol, 1.0 equiv., HCl) in MeOH (5 mL) and HO (1 mL), NaCO (13.72 mg, 129.41 μmol, 1.0 equiv.) and BocO (31.07 mg, 142.35 μmol, 32.70 μL, 1.1 equiv.) were added and stirred at 20 °C for 1 h. This mixture was added to HO (20 mL) and extracted with ethyl acetate (40 mL x 2). The organic layer was dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 100:1 to 1:2) to obtain tert-butyl N-[(5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]carbamate (85 mg, 124.66 μmol, 96.33% yield) was obtained as a yellow solid. ESI [M+ Na + ]=704.3

[0112] (ix) Preparation of Compounds 12A and 12B [ka] The racemate was purified by SFC (column: DAICEL Chiralpak AD (250 mm * 30 mm, 10 μm); mobile phase: [0.1% NH3H2O / IPA]; B%: 50% - 50%, 11 min) and arbitrarily assigned as follows: tert-Butyl N-[[(22R)-5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]carbamate (peak 1, retention time = 1.487 min, 45 mg, 66.00 μmol, 52.94% yield, ee% = 100%, yellow solid) and tert-Butyl N-[[(22S)-5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]carbamate (Peak 2, retention time = 2.137 min, 40 mg, 58.66 μmol, 47.06% yield, ee% = 98.44%, yellow solid) ESI [M+Na + ]=704.3

[0113] (x) Preparation of Compound C-2 [ka] tert-Butyl N-[[(22R)-5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ in HCl / EtOAc (1 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]carbamate (45 mg, 66.00 μmol, 1.0 equivalent) was stirred at 20 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. The obtained residue was purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 15% to 40%, 8 min) and purified with (22R)-5-amino-13-(aminomethyl)-22-hydroxy-22,24,24-trimethyl-25,25-dioxo-15-oxa-25λ.6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to obtain hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (19 mg, 30.74 μmol, 46.57% yield, 100% purity, HCl) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.40(s, 1H), 9.12(s, 1H), 8.47-8.41(m, 1H), 8.36(d, J=8.5Hz, 2H), 8.29(br s, 3H), 8.10-7.88(m, 4H), 7.35-7.28(m, 2H), 4.36-4.19(m, 1H), 4.16-4.08(m, 2H), 3.95-3.82(m, 2H), 1.86-1.73(m, 3H), 1.53(s, 3H), 1.48(s, 3H), 1.43-1.31(m, 4H), 1.28-1.10(m, 6H), 1.07-0.95(m, 2H); ESI [M+H]=582.3

[0114] Example 13: Preparation of Compound C-1 [ka] tert-Butyl N-[[(22S)-5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ in HCl / EtOAc (1 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl]methyl]carbamate (40 mg, 58.66 μmol, 1 equivalent) was stirred at 20 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. The obtained residue was purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 15% to 40%, 8 min) to obtain (22S)-5-amino-13-(aminomethyl)-22-hydroxy-22,24,24-trimethyl-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to obtain hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-7-one (16.43 mg, 26.58 μmol, 45.30% yield, 100% purity, HCl) as a yellow solid. 1 H NMR(400MHz, DMSO-d6)δ 10.39(s, 1H), 9.12(s, 1H), 8.46-8.33(m, 6H), 8.13-7.86(m, 4H), 7.38-7.27(m, 2H), 4.15-4.08(m, 2H), ESI [M+H]=582.3

[0115] Example 14: Preparation of Compound A-2 [ka]

[0116] (i) Preparation of Compound 20 [ka] 5-amino-22-hydroxy-22,24,24-trimethyl-13-(methylaminomethyl)-25,25-dioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 To a solution of hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-7-one (250 mg, 395.44 μmol, 1.0 equiv., HCl) in MeOH (10 mL) and HO (2 mL), NaCO (41.91 mg, 395.44 μmol, 1.0 equiv.) and BocO (94.93 mg, 434.98 μmol, 99.93 μL, 1.1 equiv.) were added and stirred at 20 °C for 1 h. HO (40 mL) was added and the mixture was extracted with ethyl acetate (80 mL x 2). The organic layer was dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The resulting residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 100:1 to 0:1) to obtain tert-butyl N-[(5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-13-yl)methyl]-N-methyl-carbamate (260 mg, 373.63 μmol, 94.49% yield) was obtained as a yellow solid. ESI [M+Na + ]=718.4

[0117] (ii) Preparation of Compounds 21A and 21B [ka] The racemate was purified by SFC (column: Phenomenex-Cellulose-2 (250 mm*30 mm, 10 um); mobile phase: [EtOH / ACN]; B%: 60%-60%, 20 min) and arbitrarily assigned as follows: tert-Butyl N-[[(22R)-5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-13-yl]methyl]-N-methyl-carbamate (peak 1, retention time = 1.103 min, 130 mg, 186.82 μmol, yield 50.00%, ee% = 100%, yellow solid) and tert-Butyl N-[[(22S)-5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-13-yl]methyl]-N-methyl-carbamate (peak 2, retention time = 2.602 min, 120 mg, 172.45 μmol, 46.15% yield, ee% = 100%, yellow solid) ESI [M+Na + ]=718.4

[0118] (iii) Preparation of Compound A-2 [ka] tert-Butyl N-[[(22R)-5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ in HCl / EtOAc (3 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14]Hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-13-yl]methyl]-N-methyl-carbamate (130 mg, 186.82 μmol, 1.0 equivalent) was stirred at 20 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. The obtained residue was purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 10% to 40%, 8 min) and purified with (22R)-5-amino-22-hydroxy-22,24,24-trimethyl-13-(methylaminomethyl)-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to obtain hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-7-one (79.75 mg, 126.14 μmol, 67.52% yield, 100% purity, HCl) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.39(s, 1H), 9.12(s, 3H), 8.46(dd, J=1.4, 8.1Hz, 1H), 8.37(d, J=8.6Hz, 2H), 8.15-7.83(m, 4H), 7.42-7.38(m, 1H), 7.36-7.29(m, 1H), 4.19(br t, J=5.7Hz, 2H), 3.93-3.84(m, 2H), 2.65-2.61(m, 3H), 1.87-1.72(m, 3H), 1.60(brd, J=14.1Hz, 1H), 1.54(s, 3H), 1.48(s, 3H), 1.41-1.28(m, 4H), 1.26-1.09(m, 5H), 1.07-0.90(m, 2H); ESI [M+H]=596.2

[0119] Example 15: Preparation of Compound A-1 [ka] tert-Butyl N-[[(22S)-5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15-oxa-25λ in HCl / EtOAc (3 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-13-yl]methyl]-N-methyl-carbamate (120 mg, 172.45 μmol, 1.0 equivalent) was stirred at 20 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue. The obtained residue was purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 10% to 40%, 8 min) to obtain (22S)-5-amino-22-hydroxy-22,24,24-trimethyl-13-(methylaminomethyl)-25,25-dioxo-15-oxa-25λ. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ] to obtain hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-7-one (81.45 mg, 128.83 μmol, 74.71% yield, 100% purity, HCl) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.39(s, 1H), 9.16-8.92(m, 3H), 8.46(dd, J=1.6, 7.9Hz, 1H), 8.37(d, J=8.5Hz, 2H), 8.14-7.88(m, 4H), 7.41-7.29(m, 2H), 4.20(br t, J=5.1Hz, 2H), 3.94-3.82(m, 2H), 2.64(br t, J=5.2Hz, 3H), 1.87-1.72(m, 3H), 1.60(br d, J=14.0Hz, 1H), 1.54(s, 3H), 1.48(s, 3H), 1.41-1.30(m, 4H), 1.27-1.10(m, 5H), 1.07-0.90(m, 2H); ESI [M+H]=596.2

[0120] Example 16: Preparation of Compound B-2 [ka] (i) Preparation of Compound 2 [ka] To a solution of 4-bromobenzenethiol (34 g, 179.83 mmol, 1.0 equiv) in DMSO (400 mL), t-BuOK (40.36 g, 359.65 mmol, 2.0 equiv) and bromocyclopropane (43.51 g, 359.65 mmol, 28.81 mL, 2.0 equiv) were added and stirred at 100 °C for 20 h. Ice water (1000 mL) was added to the mixture, which was then extracted with MTBE (1000 mL x 3). The organic layer was dried over Na SO , filtered, and concentrated to give 1-bromo-4-cyclopropylsulfanyl-benzene (32.5 g, crude) as a yellow oil. 1 H NMR (400MHz, methanol-d4) δ 7.31(d, J=8.5Hz, 2H), 7.17(d, J=8.5Hz, 2H), 2.17-2.04(m, 1H), 1.04-0.95(m, 2H), 0.55-0.47(m, 2H)

[0121] (ii) Preparation of Compound 3 [ka] To a solution of 1-bromo-4-cyclopropylsulfanyl-benzene (30 g, 130.93 mmol, 1.0 equiv) in MeOH (750 mL) was added a solution of oxone (185.12 g, 301.13 mmol, 2.3 equiv) in HO (750 mL) at 0 °C and stirred at 25 °C for 1 h. The mixture was concentrated to remove MeOH and then extracted with ethyl acetate (1000 mL × 2). The combined organic layers were dried over NaSO, filtered, and concentrated to give a residue. The resulting residue was triturated with MTBE (400 mL) to give 1-bromo-4-cyclopropylsulfonyl-benzene (28 g, 107.22 mmol, 81.90% yield) as a white solid. 1H NMR (400MHz, methanol-d4)δ 7.85(s, 4H), 2.73(tt, J=4.8, 7.9Hz, 1H), 1.30-1.24(m, 2H), 1.15-1.08(m, 2H); ESI [M+H]=261.0 / 263.0

[0122] (iii) Preparation of Compound 5 [ka] To a mixture of 2-(6-benzyloxyhexyl)-2-methyl-oxirane (11.41 g, 45.95 mmol, 1.2 equiv.) and 1-bromo-4-cyclopropylsulfonyl-benzene (10 g, 38.29 mmol, 1.0 equiv.) in THF (80 mL) was added LiHMDS (1 M, 57.44 mL, 1.5 equiv.) dropwise at −20° C. under a N atmosphere. The mixture was stirred at 25° C. for 12 hours under a N atmosphere. The reaction mixture was quenched with saturated aqueous NH4Cl (150 mL) at 0° C. and extracted with ethyl acetate (200 mL×2). The organic layer was dried over Na2SO4, filtered, concentrated, and purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 100:1 to 0:1) to give 8-benzyloxy-1-[1-(4-bromophenyl)sulfonylcyclopropyl]-2-methyl-octan-2-ol (9.42 g, 18.49 mmol, 48.28% yield) as a yellow oil. 1 H NMR (400MHz, chloroform-d) δ 7.78-7.72(m, 2H), 7.67-7.62(m, 2H), 7.30-7.17(m, 5H), 4.42(s, 2H), 3.97(s, 1H), 3.39(t, J=6.6Hz, 2H), ESI [M+Na + ]=531.2 / 533.2

[0123] (iv) Preparation of Compound 6 [ka] To a solution of 8-benzyloxy-1-[1-(4-bromophenyl)sulfonylcyclopropyl]-2-methyl-octan-2-ol (10 g, 19.63 mmol, 1.0 equiv) in DCM (100 mL) was added DHP (36.80 g, 437.47 mmol, 40.00 mL, 22.29 equiv) and PPTS (493.23 mg, 1.96 mmol, 0.1 equiv) and stirred at 30 °C for 12 h under a N atmosphere. HO (50 mL) was added to the mixture, which was then extracted with dichloromethane (100 mL x 2). The organic layer was dried over NaSO, filtered, and concentrated to give a residue. The resulting residue was then purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 100:1 to 0:1) to give 2-[7-benzyloxy-1-[[1-(4-bromophenyl)sulfonylcyclopropyl]methyl]-1-methyl-heptoxy]tetrahydropyran (13 g, crude) as a yellow oil. ESI [M+Na + ]=615.3 / 617.3

[0124] (v) Preparation of Compound 7 [ka] A mixture of 2-[7-benzyloxy-1-[[1-(4-bromophenyl)sulfonylcyclopropyl]methyl]-1-methylheptoxy]tetrahydropyran (13 g, 21.90 mmol, 1.0 equiv.), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (6.12 g, 24.09 mmol, 1.1 equiv.), Pd(dppf)Cl2 (1.60 g, 2.19 mmol, 0.1 equiv.), and KOAc (4.30 g, 43.80 mmol, 2.0 equiv.) in dioxane (230 mL) was stirred at 80 °C under a N2 atmosphere for 2 hours. This reaction solution was used directly in the next step. ESI [M+Na + ]=663.4

[0125] (vi) Preparation of Compound 8 [ka] tert-Butyl N-[[3-[(3-amino-6-bromo-pyrazine-2-carbonyl)amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (9.88 g, 21.85 mmol, 1.0 equiv.), 2-[4-[1-(8-benzyloxy-2-methyl-2-tetrahydropyran-2-yloxy-octyl)cyclopropyl]sulfonylphenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (14 g, 21.85 mmol, 1.0 equiv.), Na2CO3 (3.47 g, 32.78 mmol, 1.5 equiv.), Pd(dppf)Cl2 (1.60 A mixture of (g, 2.19 mmol, 0.1 equiv) in dioxane (230 mL) and HO (45 mL) was stirred at 80 °C under a N atmosphere for 12 h. The reaction mixture was concentrated, then water (200 mL) was added and extracted with ethyl acetate (200 mL × 2). The combined organic layers were dried over NaSO, filtered, and concentrated to give a residue. The residue was then purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 1:1) to give tert-butyl N-[[3-[[3-amino-6-[4-[1-(8-benzyloxy-2-methyl-2-tetrahydropyran-2-yloxyoctyl)cyclopropyl]sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxyphenyl]methyl]-N-methyl-carbamate (18.2 g, 20.54 mmol, 93.99% yield) as a yellow oil. ESI [M+H]=886.5

[0126] (vii) Preparation of Compound 9 [ka] A mixture of tert-butyl N-[[3-[[3-amino-6-[4-[1-(8-benzyloxy-2-methyl-2-tetrahydropyran-2-yloxyoctyl)cyclopropyl]sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (6.5 g, 7.34 mmol, 1.0 equiv), Pd / C (6 g, 10% purity) in MeOH (300 mL) and cyclohexene (60 mL) was stirred at 70° C. for 12 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (SiO, petroleum ether:tetrahydrofuran = 100:1 to 0:1) to give tert-butyl N-[[3-[[3-amino-6-[4-[1-(8-hydroxy-2-methyl-2-tetrahydropyran-2-yloxyoctyl)cyclopropyl]sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxyphenyl]methyl]-N-methyl-carbamate (6.2 g, crude) as a yellow solid. ESI [M+H-Boc] = 696.4

[0127] (viii) Preparation of Compound 10 [ka] To a solution of tert-butyl N-[[3-[[3-amino-6-[4-[1-(8-hydroxy-2-methyl-2-tetrahydropyran-2-yloxyoctyl)cyclopropyl]sulfonylphenyl]pyrazine-2-carbonyl]amino]-2-hydroxy-phenyl]methyl]-N-methyl-carbamate (6.2 g, 7.79 mmol, 1.0 equiv) in toluene (110 mL) was added 2-(tributyl-λ 5A solution of tert-butyl N-[(5-amino-22-methyl-7,25,25-trioxo-22-tetrahydropyran-2-yloxy-spiro[15-oxa-25λ]-2,5-dihydropyran-2-yloxy ... 6 Thia 4,8,31 Triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta (1(28),2(31),3,5,9(14),10,12,26,29-nonaene-24,1'-cyclopropan)-13-yl)methyl]-N-methyl-carbamate (5.2 g, 6.68 mmol, 85.81% yield) was obtained as a yellow solid. ESI [M+H-Boc] = 678.4

[0128] (ix) Preparation of Compound 13 [ka] 5-amino-22-hydroxy-22-methyl-13-(methylaminomethyl)-25,25-dioxo-spiro[15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14To a solution of ]hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-24,1'-cyclopropan]-7-one (180 mg, 285.63 μmol, 1.0 equiv., HCl) in MeOH (10 mL) and HO (2 mL), NaCO (30.27 mg, 285.63 μmol, 1.0 equiv.) and BocO (68.57 mg, 314.19 μmol, 72.18 μL, 1.1 equiv.) were added and stirred at 20 °C for 1 h. HO (20 mL) was added and the mixture was extracted with ethyl acetate (50 mL x 2). The organic layer was dried over NaSO, filtered, and concentrated to give a residue. The resulting residue was then purified by column chromatography (SiO2, petroleum ether:ethyl acetate=100:1 to 0:1) and purified with tert-butyl N-[(5-amino-22-hydroxy-22-methyl-7,25,25-trioxo-spiro[15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaene-24,1'-cyclopropan]-13-yl)methyl]-N-methyl-carbamate (160 mg, 230.60 μmol, 80.73% yield) was obtained as a yellow solid. ESI [M+H-Boc]=594.4

[0129] (x) Preparation of Compounds 14A and 14B [ka] The racemate was purified by SFC (column: DAICEL Chiralpak AD (250 mm * 30 mm, 10 μm); mobile phase: [0.1% NH3H2O / EtOH]; B%: 54% - 54%, 8 min) and arbitrarily assigned as follows: tert-Butyl N-[[(22R)-5-amino-22-hydroxy-22-methyl-7,25,25-trioxo-spiro[15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14]hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaene-24,1'-cyclopropan]-13-yl]methyl]-N-methyl-carbamate (peak 1, retention time = 1.484 min, 70 mg, 100.89 μmol, 43.75% yield, ee% = 100%, yellow solid) and tert-Butyl N-[[(22S)-5-amino-22-hydroxy-22-methyl-7,25,25-trioxo-spiro[15-oxa-25λ] 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-24,1'-cyclopropan]-13-yl]methyl]-N-methyl-carbamate (peak 2, retention time = 1.888 min, 90 mg, 129.71 μmol, 56.25% yield, ee% = 99.66%, yellow solid) [M+H-Boc] = 594.3

[0130] (xi) Preparation of Compound B-2 [ka] tert-Butyl N-[[(22R)-5-amino-22-hydroxy-22-methyl-7,25,25-trioxo-spiro[15-oxa-25λ] in HCl / EtOAc (2 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14]Hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-24,1'-cyclopropan]-13-yl]methyl]-N-methyl-carbamate (65 mg, 93.68 μmol, 1.0 equiv.) was stirred at 20 ° C. for 1 hour. The reaction mixture was concentrated, and the resulting residue was purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 10% to 50%, 8 min) and purified as (22R)-5-amino-22-hydroxy-22-methyl-13-(methylaminomethyl)-25,25-dioxo-spiro[15-oxa-25λ]. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaene-24,1′-cyclopropan]-7-one (31.72 mg, 50.33 μmol, 53.73% yield, 100% purity, HCl) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 10.33(s, 1H), 9.08(s, 3H), 8.37-8.28(m, 3H), 7.98(br d, J=8.3Hz, 3H), 7.42(d, J=7.8Hz, 1H), 7.35-7.29(m, 1H), 4.19(br t, J=5.1Hz, 3H), 3.91-3.76(m, 2H), 2.64(br t, J=4.9Hz, 3H), 2.09-2.01(m, 1H),1.97-1.90(m, 1H), 1.81-1.69(m, 1H), 1.68-1.54(m, 3H), 1.53-1.43(m, 2H), 1.36(dt, J=6.3, 11.7Hz, 1H), 1.28-1.17(m,1H), 1.15-1.00(m, 2H), 0.99-0.67(m, 7H); ESI [M+H]=594.3

[0131] Example 17: Preparation of Compound B-1 [ka] tert-Butyl N-[[(22S)-5-amino-22-hydroxy-22-methyl-7,25,25-trioxo-spiro[15-oxa-25λ] in HCl / EtOAc (2 mL, 4 M) 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]Hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaen-24,1'-cyclopropan]-13-yl]methyl]-N-methyl-carbamate (85 mg, 122.50 μmol, 1.0 equiv.) was stirred at 20 ° C. for 1 hour. The reaction mixture was concentrated, and the resulting residue was purified by preparative HPLC (column: Phenomenex Luna 80*30 mm*3 μm; mobile phase: [water (HCl)-ACN]; B%: 10% to 50%, 8 min) to give (22S)-5-amino-22-hydroxy-22-methyl-13-(methylaminomethyl)-25,25-dioxo-spiro[15-oxa-25λ]. 6 -Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2,4,6(31),9(14),10,12,26,29-nonaene-24,1′-cyclopropan]-7-one (37.42 mg, 59.38 μmol, 48.47% yield, 100% purity, HCl) was obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ 10.33(s, 1H), 9.09(s, 1H), 8.96(br d, J=5.8Hz, 2H), 8.40-8.27(m, 3H), 7.99(br d, J=8.4Hz, 3H), 7.42-7.29(m, 2H), 4.28-4.15(m, 3H), 3.90-3.76(m, 2H), 2.65(br t, J=5.1Hz, 3H), 2.08-2.01(m, 1H), 1.98 -1.89(m, 1H), 1.83-1.70(m, 1H), 1.67-1.55(m, 3H), 1.53-1.42(m, 2H), 1.41-1.30(m, 1H), 1.27-1.17(m, 1H), 1.16-1.02(m, 2H), 0.89(s, 7H); ESI [M+H]=594.3

[0132] Example 18: Preparation of Compound J-1 [ka]

[0133] (i) Preparation of Compound 2 [ka] To a solution of tert-butyl N-[(32-amino-36,36,37-trimethyl-34,53,53-trioxo-37-tetrahydropyran-2-yloxy-48,49-dioxa-53thia-40,41,42-triazatetracyclohentriaconta-6(8),7(27),9(25),10(12),11(26),13(40),28(30),29(41),31-nonaen-27-yl)methyl]carbamate (120 mg, 156.26 μmol, 1.0 equiv.) in EtOH (5 mL) was added TsOH·HO (5.94 mg, 31.25 μmol, 0.2 equiv.) and stirred at 30 °C for 1 h. The reaction mixture was concentrated and tert-butyl N-[(5-amino-22-hydroxy-22,24,24-trimethyl-7,25,25-trioxo-15,20-dioxa-25λ] 6-Thia-4,8,31-triazatetracyclo[24.2.2.1 2,6 .0 9,14 ]hentriaconta-1(28),2(31),3,5,9(14),10,12,26,29-nonaen-13-yl)methyl]carbamate (110 mg, 160.86 μmol, 102.94% yield) was obtained as a yellow solid. ESI [M+H]=684.3

[0134] (ii) Preparation of Compounds 3A and 3B [ka] The racemate was purified by SFC (column: DAICEL Chiralpak IC (250 mm * 50 mm, 10 um); mobile phase: [0.1% NH3H2O / MeOH]; B%: 50% - 50%, 40 min) and arbitrarily assigned as follows: tert-Butyl N-[[(32S)-28-amino-32-hydroxy-31,31,32-trimethyl-29,47,47-trioxo-44,45-dioxa-47thia-35,36,37-triazatetracyclohentriaconta-6(8),7(23),9(21),10(12),11(22),13(35),24(26),25(36),27-nonaen-23-yl]methyl]carbamate (peak 1, retention time = 3.89 min, 50 mg, 73.12 μmol, yield 93.58%, ee% = 100%, yellow solid) and tert-Butyl N-[[(32R)-28-amino-32-hydroxy-31,31,32-trimethyl-29,47,47-trioxo-44,45-dioxa-47thia-35,36,37-triazatetracyclohentriaconta-6(8),7(23),9(21),10(12),11(22),13(35),24(26),25(36),27-nonaen-23-yl]methyl]carbamate (Peak 2, Retention time = 6.16 min, 50 mg, 73.12 μmol, Yield 93.58%, ee% = 100%, Yellow solid) ESI [M+H] = 684.3

[0135] (iii) Preparation of Compound J-1 [ka] A solution of tert-butyl N-[[(32S)-28-amino-32-hydroxy-31,31,32-trimethyl-29,47,47-trioxo-44,45-dioxa-47thia-35,36,37-triazatetracyclohentriaconta-6(8),7(23),9(21),10(12),11(22),13(35),24(26),25(36),27-nonaen-23-yl]methyl]carbamate (50 mg, 73.12 μmol, 1.0 equiv) in HCl / MeOH (1 mL, 4 M) was stirred at 30 °C for 0.5 h. The reaction mixture was concentrated, and the resulting residue was purified by preparative HPLC (column: Phenomenex Luna C18 100*30mm*5μm; mobile phase: [water (0.04% HCl)-ACN]; B%: 15%-45%, 10 min) to obtain (28S)-25-amino-20-(aminomethyl)-28-hydroxy-27,27,28-trimethyl-40,40-dioxo-38,39-dioxa-40thia-31,32,33-triazatetracyclohentriaconta-3(5),4(20),6(18),7(9),8(19),10(31),21(23),22(32),24-nonaen-26-one (29.3 mg, 47.13 μmol, yield 64.46%, purity 99.76%, HCl) as a yellow solid. 1H NMR(400MHz, DMSO-d6) δ 10.44(s, 1H), 9.05(s, 1H), 8.61(br s, 3H), 8.48-8.41(m, 1H), 8.28(d, J=8.4Hz, 2H), 7.89(d, J=8.4Hz, 3H), 7.39(d, J=7.7Hz, 1H), 7.33-7.23(m, 1H), 4.09(br d, J=5.4Hz, 2H), 3.96(br d, J=10.1Hz, 2H), 3.41-3.30(m, 1H), 3.27-3.16(m, 1H), 3.15-3.01(m, 2H), 1.98(br d, J=14.3Hz, 1H), 1.91-1.80(m, 2H), 1.73(br d, J=14.2Hz, 1H), 1.77-1.69(m, 1H), 1.67-1.55(m, 2H), 1.49(br d, J=12.2Hz, 6H), 1.12(s, 3H); ESI [M+H]=584.3

[0136] Manufacturing of Compound J-2

change

[0137] Example 20: Assay A. Cell seeding Cells were seeded for a specific assay or set of assays. Different cell lines require different numbers of cells per cm due to the different surface areas per cell. The number of cells / cm required for each assay and cell line is 2 is shown in Table 1. [Table 1] The cells to be seeded are passaged two or three times after thawing. 1. Preheat the cell culture medium in a 37°C water bath and add 0.05% or 0.25% trypsin. 2. The cells to be seeded were observed under bright field at 10x and 40x magnification for their apparent health and cell coverage. Cells should be seeded when they are in the logarithmic growth phase, approximately 80% confluent. 3. Aspirate the cell culture medium from the flask and add 75 cm 2 The cells were washed twice with 15 mL (for flasks) or 10 mL (for 10 cm cell culture dishes) of 1x PBS. 4. 0.05% or 0.25% trypsin medium (1.5 mL) was added, and the flask / dish was tilted so that all cells were in contact with the trypsin medium. The flask / dish was returned to the incubator and left to stand for 5 minutes. 5. To inactivate trypsin while detaching the cells from the surface of the Petri dish, add three times the volume of pre-warmed cell culture medium (4.5 mL) of the trypsin added above. Carefully pipette the cells to wash them off the surface. The total volume of the cell suspension was 6 mL. 6. (When multiple plates were used) The cell suspensions from all plates were combined and collected in a 50 mL centrifuge tube. 7. 40 μL of the cell suspension was removed and added to trypan blue (40 μL) in an Eppendorf tube (1.5 mL). 8. The cell suspension / dye mixture (10 μL) was injected into each end of the hemocytometer, and the number of cells was counted using an automated cell counter. 9. The number of viable cells was recorded for each block of the hemocytometer. If the viability counts were greater than 95% and within 10% of each other, the counts were averaged to calculate the total cell count. If these criteria were not met, this step was repeated. If the criteria were again not met, step 8 was repeated. The remaining cell suspension / dye mixture was discarded. 10. The total number of cells in the cell suspension was calculated as follows: Total cell number = (number of cells / mL from step 9) * (total volume of cell suspension (mL)) 11. The number of cells required for the experiment was calculated as follows: Required number of cells = (number of wells to be seeded + 2) * (seeding density / well in Table 1) 12. The volume of cell suspension required for the experiment was calculated as follows: (Number of cells required) / (Cell density in step 9) = Required cell suspension (mL) 13. The volume of cell suspension required in step 12 was gently pelleted in a 50 mL centrifuge tube (1000 rpm, 5 minutes). 14. The volume of medium needed to resuspend the cell pellet was calculated as follows: Medium (mL) = number of wells to be seeded 15. The cells were pelleted, the supernatant removed and resuspended in the calculated volume of pre-warmed cell culture medium. 16. Freshly pre-warmed cell culture medium (1 mL) was added to each well of an appropriate number of sterile, labeled 6-well plates to be used for seeding. In the standard exposure method, two 6-well plates are required per compound. 17. The cell resuspension (1 mL) was added to each well. The tube containing the cell suspension was gently swirled or pipetted to prevent the cells from settling at the bottom of the tube. The plate was gently shaken in a figure-eight motion to distribute the cells evenly throughout the wells. 18. The remaining cell suspension was replated at the appropriate density for later use. 19.Atrize TMFor the ELISA and cell proliferation assays, compounds can be exposed immediately. For all other assays, cells can be incubated for 24 hours before compound exposure. 20. The plate was incubated (37°C, 5% CO2).

[0138] B. Cell Proliferation Assay - Range Determination (i) Cell seeding and treatment (day 0) The cell culture medium was preheated in a 37°C water bath and 0.05% trypsin was added. Compound stock solutions were prepared at a concentration of 1 mM. Compounds with low pharmacological activity (dosed at >100 nM) did not require further dilution and were used at the stock concentration. For compounds with high pharmacological activity (dosed at <100 nM), the compound was diluted 1:10 in DMSO in an Eppendorf tube [DMSO (90 μL) + 1 mM compound stock solution (10 μL)] to a concentration of 100 μM. The diluted, highly pharmacologically active compound was divided into five compound working aliquots (20 μL). The aliquots were stored at -20°C. A new aliquot was used for counting and re-plating the cells at a later date. The dilution factor was 2x the final concentration. [Table 2] The following formula (based on CC1VV1=CC2VV2) was used to determine the volume of compound stock solution or compound working aliquot to add to the Spiking Solution: Volume of 1 mM compound stock solution to add to 8 mL of highest concentration medium: [ka] Volume of 100 μM compound working aliquot when added to highest concentration medium (8 mL): [ka] To perform the range-finding assay, 18 wells (three 6-well plates) per drug compound (drug) were required for three identical replicates. The appropriate loading solution (1 mL) was added to each well and the plate was placed in an incubator. The cell culture medium was removed from the dish / flask by suction and washed twice with 5 mL of 1x PBS. 1.5 mL of 0.05% trypsin medium was added, and the dish / flask was tilted so that all cells were in contact with the trypsin medium. The flask / dish was returned to the incubator and left undisturbed for 5 minutes. To inactivate trypsin while detaching cells from the surface of the dish, pre-warmed cell culture medium was added in a volume three times the volume of trypsin added above (4.5 mL). The cells were washed off the surface by careful pipetting. The total volume of the cell suspension (per dish / flask) was 6 mL. When multiple dishes were used, the cell suspensions from all dishes were combined.

[0139] Trypan blue (1 mL) was centrifuged at maximum speed for 5 minutes. The supernatant was transferred to an Eppendorf tube in a safety cabinet before use. 20 μL of the cell suspension was removed and added to 20 μL of trypan blue in a 1.0 mL Eppendorf tube. 10 μL of the cell suspension / dye mixture was injected into each end of a hemocytometer, and the cells were counted twice (two samples) using an automated cell counter. The number of viable cells was recorded for each block of the hemocytometer. If the counted viability was greater than 90% and within 10% of each other, the counts were averaged. If these criteria were not met, this step was repeated. Cells were prepared according to the following steps: 1. The total number of cells in the cell suspension was calculated as follows: Total cell count = (number of cells / mL) * (total volume of cell suspension (mL)) 2. The number of cells required for the experiment was calculated as follows: Required number of cells = (number of wells to be seeded + 2) * (seeding density / well in Table 3) [Table 3] 3. The volume of cell suspension required for the experiment was calculated as follows: (Number of cells required) / (Average cell density on hemocytometer) = Required cell suspension (mL) 4. The volume of cells in suspension required in step 3 was pelleted into a 50 mL centrifuge tube (1000 rpm, 5 minutes). 5. The volume of medium needed to resuspend the cell pellet was calculated as follows: Medium (mL) = number of wells to be seeded + 2 6. Pellet the cells in a 50 mL centrifuge tube, remove the supernatant, and resuspend in pre-warmed cell culture medium (volume calculated in step 5). The tube containing the cell suspension was gently swirled to disperse the cells in the medium. On day 0 only, the cell resuspension (1 mL) was added to each of the 18 wells. Note that on days 2 and 4, the volume of cell suspension (and the additional fresh medium) added to the wells will vary depending on the calculation. The plate was then placed in the incubator.

[0140] (ii) Counting and Replating (Days 2 and 4) Preheat the cell culture medium in a 37°C water bath and add 0.05% or 0.25% trypsin. Prepare the additive solution as described on Day 0. Remove the cell culture medium from the wells by aspiration and wash once with 2 mL of 1x PBS. Add 250 μL of 0.05% trypsin medium and tilt the dish so that all cells are exposed to the trypsin medium. Return the flask / dish to the incubator and let it sit for 5 minutes. To inactivate trypsin while detaching the cells from the surface of the Petri dish, pre-warmed cell culture medium was added in a volume three times the volume of trypsin added above (750 μL). The cells were washed off the surface by careful pipetting. The total volume of the cell suspension was 1 mL per well. The contents of each well were transferred to an appropriate 1.5 mL tube. For each tube, 30 μL of cell suspension was removed and added to 30 μL of trypan blue in a correspondingly labeled Eppendorf tube (1.0 mL). 10 μL of the cell suspension / dye mixture was injected through the edge of the hemocytometer, and the cells were counted using an automated cell counter. This process was repeated for each well / tube. If there were not enough cells to meet the seeding density in Table 1, back-calculate the number of cells to seed per well and return ∼850 μL of cell suspension (along with 150 μL of fresh medium) to the well. The spreadsheet formula was adjusted to reflect the actual number of cells seeded. All three wells under the same seeding condition should have the same cell number. Loading solution, cell suspension, and fresh medium were added to the wells as calculated.

[0141] (iii) Counting only (day 6) or counting and replating as needed (day 6 or 8) Each well was considered to be an independent experiment, and cell suspensions were not pooled at any time. Cell numbers were counted in the same manner as described in (ii). If significant death was observed in the high-dose group, the cell suspension was discarded.

[0142] (iv) Formula The cell proliferation number on day 2 was calculated as follows: [ka] *"Cell count" is the average of the cell counts on the second day for three cells under the same addition conditions. The subsequent cell proliferation was calculated as follows: [ka] *"Cell count" is the average of three cell counts under the same addition conditions on the relevant day, and "previous count" is the average of three cell counts under the same addition conditions on the previous day.

[0143] C. Cell proliferation assay (two drugs) (i) Cell seeding and treatment (day 0) Compound stock solutions were prepared at a concentration of 1 mM. Compounds with low pharmacological activity (dosed at >100 nM) did not require further dilution and were used at the stock concentration. For compounds with high pharmacological activity (dosed at <100 nM), the compound was diluted 1:10 in DMSO in an Eppendorf tube [DMSO (90 μL) + 1 mM compound stock solution (10 μL)] to a concentration of 100 μM. The diluted pharmacologically active compounds were divided into five compound working aliquots (20 μL). The aliquots were stored at -20°C. A new aliquot was used for counting and re-plating the cells at a later date. Each compound was diluted in 4 mL of cell culture medium to provide a 4-fold dilution of the final concentration of compound added to the cells. The compound diluted in cell culture medium (1 mL) constituted 25% of the volume of each well. See Table 4 for the final composition of the wells. Depending on the amount added, compounds as listed in Table 4 or DMS as a vehicle and cell culture medium were added to three centrifuge tubes (15 mL). The cell culture medium was preheated in a 37°C water bath and 0.05% trypsin was added. [Table 4] The following formula (based on CC1VV1=CC2VV2) was used to determine the volume of compound stock solution or compound working aliquot to add to the Spiking Solution: Volume of 1 mM compound stock solution to add to 4 mL of medium: [ka] Volume of 100 μM compound working aliquot when added to medium (4 mL): [ka] To perform the assay for two drug compounds (drugs), 12 wells (two 6-well plates) were required for three identical replicates. Addition solutions and fresh medium were added to the wells as described in Table 5, and the plates were then incubated. [Table 5] The cell culture medium was removed from the dish / flask by suction and washed twice with 1x PBS (5 mL). 0.05% trypsin medium (1.5 mL) was added, and the dish / flask was tilted so that all cells were in contact with the trypsin medium. The flask / dish was returned to the incubator and left to stand for 5 minutes. To inactivate trypsin while detaching cells from the surface of the dish, pre-warmed cell culture medium was added in a volume three times the volume of trypsin added above (4.5 mL). The cells were washed off the surface by careful pipetting. The total volume of the cell suspension (per dish / flask) was 6 mL. When multiple dishes were used, the cell suspensions from all dishes were combined.

[0144] Trypan blue (1 mL) was centrifuged at maximum speed for 5 minutes. The supernatant was transferred to an Eppendorf tube in a safety cabinet before use. 20 μL of the cell suspension was removed and added to trypan blue (20 μL) in an Eppendorf tube (1.0 mL). The cell suspension / dye mixture (10 μL) was injected into each end of the hemocytometer, and the cells were counted twice (two samples) using an automated cell counter. The number of viable cells was recorded for each block of the hemocytometer. If the counted viability was greater than 90% and within 10% of each other, the counts were averaged. If these criteria were not met, this step was repeated. The remaining cell suspension / dye mixture was discarded. The cells were prepared according to the following steps: 1. The total number of cells in the cell suspension was calculated as follows: Total cell number = (number of cells / mL) * (total volume of cell suspension (mL)) 2. The number of cells required for the experiment was calculated according to Table 3. Required number of cells = (number of wells to be seeded + 2) * (seeding density / well in Table 3) 3. The volume of cell suspension required for the experiment was calculated as follows: (Number of cells required) / (Average cell density on hemocytometer) = Required cell suspension (mL) 4. The volume of cells in suspension required in step 3 was pelleted into a 50 mL centrifuge tube (1000 rpm, 5 minutes). 5. The volume of medium needed to resuspend the cell pellet was calculated as follows: Medium (mL) = number of wells to be seeded + 2 6. Pellet the cells in a 50 mL centrifuge tube, remove the supernatant, and resuspend in pre-warmed cell culture medium (volume calculated in step 5). The tube containing the cell suspension was gently agitated to disperse the cells in the medium. On day 0 only, the cell resuspension (1 mL) was added to each of 12 wells. On days 2 and 4, the volume of cell suspension (and fresh medium) added to the wells was calculated according to the values ​​in Table 3. The plate was placed in an incubator.

[0145] (ii) Counting and replating (days 2 and 4, and day 6 as appropriate) Preheat the cell culture medium in a 37°C water bath and add 0.05% or 0.25% trypsin. Each well is considered a separate experiment; cell suspensions may not be pooled at any time. The cell culture medium was aspirated from the wells and washed once with 1x PBS (2 mL). 250 μL of 0.05% trypsin medium was added, and the dish was tilted so that all cells were in contact with the trypsin medium. The flask / dish was returned to the incubator and left undisturbed for 5 minutes. To inactivate trypsin while detaching the cells from the surface of the dish, three times the volume of pre-warmed cell culture medium (750 μL) was added. The cells were washed off the surface by gentle pipetting. The total volume of the cell suspension was 1 mL per well. The contents of each well were transferred to an appropriately labeled 1.5 mL tube. For each tube, 30 μL of cell suspension was removed and added to 30 μL of trypan blue in a correspondingly labeled Eppendorf tube (1.0 mL). 10 μL of the cell suspension / dye mixture was injected through the edge of the hemocytometer, and the cells were counted using an automated cell counter. This process was repeated for each well / tube. If there were not enough cells to meet the seeding density in Table 1, the number of cells to seed per well was back-calculated and ∼850 μL of cell suspension (along with 150 μL of fresh medium) was added back to the well. The spreadsheet formula was adjusted to reflect the actual number of cells seeded. Three wells with identical loading conditions were calculated and should have the same cell number. Loading solution, cell suspension, and fresh medium were added to the wells as calculated.

[0146] (iii) Counting only or counting and replating as needed (day 6 or 8) Each well was considered a separate experiment, and cell suspensions were not pooled at any time. Cell numbers were counted using the same method as described in (ii). These results were entered into a spreadsheet created for the experiment. If there was no significant decrease in proliferation in the wells where Compound 1 and Compound 2 were combined, the cell numbers were entered into the growth curve in the spreadsheet, the tube containing the cell suspension was incubated, and the additive solution described on Day 0 was prepared. Other procedures were then carried out on Days 2 and 4. If significant cell death was observed in the combination group, the cell suspension was discarded.

[0147] (iv) Formula The cell proliferation number on day 2 was calculated as follows: [ka] "Cell count" is the average of the cell counts on the second day for three cells under the same addition conditions. The subsequent cell proliferation was calculated as follows: [ka] "Cell count" is the average of three cell counts under the same addition conditions on the relevant day, and "previous count" is the average of three cell counts under the same addition conditions on the previous day.

[0148] D. Protein Quantification A BSA standard solution was prepared and aliquoted. 1. Remove the prepared BSA standard solution from the freezer and allow it to warm to room temperature. Vortex each aliquot briefly. 2. Remove the collected treated cell samples from the freezer and allow them to warm to room temperature. Vortex each sample briefly. 3. Standard solutions and samples were added to a 96-well flat-bottom plate according to the following steps. 4. BSA standard solution was added. 25 μL of BSA standard solution (1500 μg / mL) was added to wells A1 and B1, respectively. 25 μL of BSA standard solution (1000 μg / mL) was added to wells A2 and B2, respectively. 25 μL of BSA standard solution (750 μg / mL) was added to wells A3 and B3, respectively. 25 μL of BSA standard solution (500 μg / mL) was added to wells A4 and B4, respectively. 25 μL of BSA standard solution (250 μg / mL) was added to wells A5 and B5, respectively. 25 μL of BSA standard solution (125 μg / mL) was added to wells A6 and B6, respectively. 25 μL of BSA standard solution (25 μg / mL) was added to wells A7 and B7, respectively. 25 μL of BSA standard solution (0 μg / mL [blank]) was added to wells A8 and B8, respectively. 5. Add the harvested cell samples (row D is identical to row C). The steps below list the concentrations shown in the standard addition scheme. If a different addition method is used, adjust accordingly. Note that for untreated APH+ samples, the treated addition range is combined. · 25 μL of sample (NT) was added to wells C1 and D1, respectively. 25 μL of sample (APH+) was added to wells C2 and D2, respectively. 25 μL of sample (100 nM) was added to wells C3 and D3, respectively. 25 μL of sample (30 nM) was added to wells C4 and D4, respectively. 25 μL of sample (10 nM) was added to wells C5 and D5, respectively. 25 μL of sample (3 nM) was added to wells C6 and D6, respectively. 25 μL of sample (1 nM) was added to wells C7 and D7, respectively. 25 μL of sample (300 pM) was added to wells C8 and D8, respectively. 25 μL of sample (100 pM) was added to wells C9 and D9, respectively. 25 μL of sample (30 pM) was added to wells C10 and D10, respectively. 25 μL of sample (10 pM) was added to wells C11 and D11, respectively. 25 μL of sample (APH+) was added to wells C12 and D12, respectively. 6. The volume of Reagent A in the kit: Calculate the number of required wells x 200 μL + 10%, and add this amount to a 15 mL centrifuge tube. 7.1 / 50 (i.e., 0.02x) volume of Reagent B from the kit was added to the centrifuge tube and mixed well by inversion. 8. The reagent mixture was added to the reagent well. 9. The reagent mixture (200 μL) was transferred to each well of the standard solution and sample in the 96-well plate. Mix well by pipetting. 10. The plate was covered. 11. Incubated for 30 minutes. 12. Remove from incubator and allow to return to room temperature in a safety cabinet. 13. The results were analyzed using a microplate reader.

[0149] E. IC by Western blotting 50 Cell processing for assay Compounds were stored as 1 mM stock solutions and prepared according to the compound SOP method. The stock solutions were diluted in two separate 1:10 dilution series into cell culture medium. The dilution series of the added compounds are listed in Tables 6 and 7. [Table 6] [Table 7]

[0150] 1. Twelve 15 mL centrifuge tubes were arranged in two rows on a rack and labeled according to dilution series 1 and dilution series 2 shown in Tables 1 and 2, respectively. 2. Pre-warmed cell culture medium (2.5 mL) was added to the 100 nM tube (series 1) and the 300 nM tube (series 2), respectively. Pre-warmed cell culture medium (2.25 mL) was added to the remaining tubes. 3. 7.5 μL of 1 mM compound stock solution was added to the 3 μM tube and mixed well. 4. 2.5 μL of 1 mM compound stock solution was added to the 1 μM tube and mixed well. 5. For each series (separately), remove 250 μL from the left-most tube and transfer to the adjacent tube on the right. Repeat this for each tube until the series is complete. 6. The 1 μM, 3 μM, and 300 nM tubes were set aside for disposal. 7. The cells to be treated were seeded into 6-well plates labeled for each cell line. The cells were removed from the incubator and labeled with the dose to be added. 8. The dilution tubes were arranged in a rack to correspond to the layout of the 6-well plate to avoid confusion and ensure that the solutions were added to the appropriate wells. 9. The three wells in the top row of the first plate (NT, APH+, and APH+) were aspirated and replaced with 2 mL of fresh medium (without drug compound). 10. Aspirate the three wells in the bottom row of the first plate and replace each well with 2 mL of solution from the appropriate tube. 11. Step 10 was repeated with the wells in the next row (top row of the second plate) and then the last row (bottom row of the second plate). 12. Both plates were incubated for 30 minutes. 13. After 30 minutes, the plate was removed from the incubator. 14. APH stock solution (5 mM) was prepared and added (2 μL) to the center of each well except for the well labeled NT. 15. The plate was carefully and gently agitated to distribute the APH throughout each well. 16. The plates were again incubated for 4 hours and then the cells were harvested for analysis. 17. 20 minutes before cell collection (or 3 hours and 40 minutes after APH administration), start heating the heat block to heat the Eppendorf tubes to the appropriate temperature. The temperature was set to 95°C. 18. Prepare a number of Eppendorf tubes equal to the number of wells to be treated (i.e., 12 for two drug addition plates) and label each tube with the cell line, compound added, dose, and date. Leave the tube caps open. 19. Ion-exchanged water (200 mL) was added to a beaker. 20. After 4 hours, the treated plates were removed from the incubator. 21. The medium was aspirated from all wells. 22. Each well was washed with 1x PBS (2 mL) and the solution was removed by aspiration. 23. Spray the vacuum nozzle with 70% ethanol to sterilize the vacuum line. Turn off the vacuum. 24. Laemmli buffer was added to each well: 300 μL / well for HCT116 Bcl / xl, OVCAR-3, and 22RV1 cells, and 250 μL / well for OVCAR-8 and SKOV-3 cells. 25. The first well was completely sampled and placed in deionized water for use. 26. Using a 1000 μL pipette, carefully transfer the lysed cell debris mixture into the original tube. This mixture was viscous. The tube was capped. 27. Steps 25 and 26 were repeated for each well. 28. When sampling all wells, all tubes were placed on a heat block. A heavy, heat-resistant object was placed on top of the tubes to keep the lids closed. 29. After 5 minutes, remove the tubes from the heat block and turn it off. Vortex each tube for 10 seconds, taking care not to pop the tubes open. 30. The tubes were allowed to come to room temperature and then analyzed or frozen at -20°C for storage.

[0151] F. Results Compound X was synthesized as described in US Pat. No. 9,663,535, incorporated herein by reference. VE822 and AZD6738 are ATR inhibitors used in the field. [Table 8-1] [Table 8-2] [Table 8-3]

[0152] While the present invention has been described in conjunction with its preferred specific embodiments, it should be understood that the foregoing description and the examples that follow are illustrative of the scope of the invention and are not intended to limit it. It will be understood that various modifications and substitutions of equivalents may be made without departing from the invention, and that other aspects, advantages, and modifications will be apparent to those skilled in the art to which the invention pertains. In addition to the embodiments described herein, the present disclosure relates to and claims inventions that result from combinations of features of the invention referred to herein and features of the cited prior art documents that supplement features of the invention. Similarly, it is understood that any described substance, feature, or item may be used in combination with any other substance, feature, or item, and such combinations are considered to be within the scope of the invention.

[0153] The disclosures of each patent, patent application, and publication cited or described in this application are incorporated herein in their entirety for all purposes.

Claims

1. Formula (I): 【Chemistry 1】 [In the formula, R 1 is H, C 1-6 Alkyl or substituted C 1-6 is alkyl; R 2 is H, C 1-6 Alkyl or substituted C 1-6 is alkyl, or R 1 and R 2 are C which may be substituted together as appropriate 3-6 forms a cycloalkyl or optionally substituted heterocycloalkyl; R 3 is H, C 1-6 Alkyl or substituted C 1-6 is alkyl; R 4 , R 5 , R 6 , and R 7 are independently H, C 1-6 Alkyl or substituted C 1-6 is alkyl; L is C 1-20 alkylene, where C 1-20 One or more carbon atoms of the alkylene may each be optionally replaced by an oxygen atom. or a pharmaceutically acceptable salt thereof.

2. In the formula, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , and R 7 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein at least one of is H.

3. In the formula, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , and / or R 7 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: is independently selected from methyl, ethyl, propyl, butyl, pentyl, or hexyl.

4. In the formula, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , and / or R 7 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is independently selected from substituted methyl, substituted ethyl, substituted propyl, substituted butyl, substituted pentyl, or substituted hexyl.

5. In the formula, R 1 and R 2 However, together they form unsubstituted or substituted C 3-6 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, which forms a cycloalkyl.

6. In the formula, R 1 and R 2 and R 1 and R 2 are independently substituted or unsubstituted heterocycloalkyl; and R 3 and R 4 are independently substituted or unsubstituted heterocycloalkyl; and R 5 and R 6 are independently substituted or unsubstituted heterocycloalkyl; and R 6 and R 7 are independently substituted or unsubstituted heterocycloalkyl; and R 8 and R 9 are independently substituted or unsubstituted heterocycloalkyl; and R 10 and R 11 and R 12 and R 13 and R 14 and R 15 and R

7. In the formula, R 1 and R 2 and R are taken together to form unsubstituted or substituted pyrrolidinyl, unsubstituted or substituted tetrahydrofuranyl, unsubstituted or substituted oxetanyl, unsubstituted or substituted pyranyl, unsubstituted or substituted piperidinyl, or unsubstituted or substituted azetidinyl, or a pharmaceutically acceptable salt thereof.

8. In the formula, R 4 , R 5 , R 6 , R 7 , and / or R 8 But independently, NH 2 , NH(C 1-6 alkyl), or N(C 1-6 Alkyl)(C 1-6 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is substituted with (alkyl).

9. In the formula, R 4 , R 5 , R 6 , R 7 , and / or R 8 But independently, NHCH 3 , NHCH 2 CH 3 , N(CH 3 ) 2 , N(CH 2 CH 3 ) 2 , or N.H. 2 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, substituted with:

10. wherein L is unsubstituted C 1-20 Alkylene, unsubstituted C 1-18 Alkylene, unsubstituted C 1-16 Alkylene, unsubstituted C 1-14 Alkylene, unsubstituted C 1-12 Alkylene, unsubstituted C 1-10 Alkylene, unsubstituted C 1-8 Alkylene, unsubstituted C 1-6 Alkylene or unsubstituted C 1-4 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

11. wherein L is a substituent C 1-20 Alkylene, substituted C 1-18 Alkylene, substituted C 1-16 Alkylene, substituted C 1-14 Alkylene, substituted C 1-12 Alkylene, substituted C 1-10 Alkylene, substituted C 1-8 Alkylene, substituted C 1-6 Alkylene, or substituted C 1-4 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

12. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein L contains 1 to 6 oxygen atoms.

13. The chemical formula is 【Chemistry 2】 【Transformation 3】 【Chemistry 4】 2. The compound of claim 1, wherein:

14. 14. The compound of any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein the salt is an HCl salt.

15. A pharmaceutical composition comprising one or more compounds according to any one of claims 1 to 13 or pharmaceutically acceptable salts thereof.

16. A pharmaceutical composition for treating cancer, comprising the compound of any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof.

17. The cancer in question is brain tumor, breast cancer, central nervous system cancer, colorectal cancer, glioblastoma, melanoma, leukemia, liver cancer, lung cancer, lymphoma, pancreatic cancer, prostate cancer, ovarian cancer, kidney cancer, melanoma of the eye, desmoplastic round cell tumor, chondrosarcoma, leptomeningeal disease, diffuse large B-cell lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancer, AIDS-related lymphoma, anal or rectal cancer, appendix cancer, astrocytoma, atypical teratoid rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, osteosarcoma or malignant fibrous histiocytoma, bronchial tumor, Burkitt's disease, or rectal cancer. Lymphoma, spinal cord tumor, carcinoid tumor, cancer of unknown primary site, atypical teratoid rhabdoid tumor of the central nervous system, leptomeningeal disease, central nervous system embryonal tumor, central nervous system lymphoma, chordoma, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloproliferative disorder, colon cancer, craniopharyngioma, cutaneous T-cell lymphoma, ependymoblastoma, ependymoma, esophageal cancer, Ewing's sarcoma family of tumors, extracranial germ cell tumor, extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancer, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, glioma, hairy cell leukemia, head and neck cancer, hepatocellular carcinoma alveolar carcinoma, histiocytosis, Hodgkin's lymphoma, hypopharyngeal cancer, Kaposi's sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, lip or oral cavity cancer, non-Hodgkin's lymphoma, primary central nervous system malignant lymphoma, Waldenstrom's macroglobulinemia, medulloblastoma, medulloepithelioma, Merkel cell carcinoma, mesothelioma, metastatic squamous cell carcinoma of the neck of unknown primary, multiple endocrine neoplasia, mouth cancer, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, myeloproliferative disorders, nasal cavity or paranasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma, oral cavity cancer, oropharyngeal cancer, osteosarcoma or bone marrow cancer malignant fibrous histiocytoma, papilloma, paranasal sinus or nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, intermediate pineal parenchymal tumor, pineoblastoma or supratentorial primitive neuroectodermal tumor, pituitary tumor, pleuropulmonary blastoma, rectal cancer, pelvic or ureteral cancer, respiratory cancer associated with the NUT gene on chromosome 15, retinoblastoma, rhabdomyosarcoma, high-grade prostate cancer, intermediate-grade prostate cancer, low-grade prostate cancer, castration-resistant prostate cancer, salivary gland cancer, non-epithelial malignant tumor, Sezary syndrome, skin cancer, skin malignant tumor, eye tumor, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous cell carcinoma of the neck of unknown primary origin,17. The pharmaceutical composition according to claim 16, wherein the cancer is selected from supratentorial primitive neuroectodermal tumor, T-cell lymphoma, testicular cancer, pharyngeal cancer, thymoma or thymic carcinoma, thyroid cancer, transitional cell carcinoma of the kidney or renal pelvis or ureter, cancer of unknown primary, epithelial malignant tumor of unknown primary, rare cancer of childhood, urethral cancer, Wilms' tumor, or gynecological cancer selected from cervical cancer, endometrial cancer, gestational trophoblastic tumor, ovarian germ cell tumor, epithelial ovarian cancer, ovarian low malignant potential tumor, gestational cancer, uterine sarcoma, vaginal cancer, and vulvar cancer.

18. 16. The pharmaceutical composition of claim 15, which is administered orally.

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