Novel compounds for the degradation of targeted proteins or polypeptides by polyubiquitination

Abstract

The present invention relates to a compound capable of degrading a target protein using ubiquitin, and provides a compound having a TBM (targeted protein binding moiety) structure that binds to a target protein or polypeptide and a UBM (ubiquitin binding moiety) structure that binds to ubiquitin. The compound of the present invention has an advantage that it can become a protein degrader that can degrade a target protein or polypeptide, since polyubiquitination is induced by the UBM structure present in the compound.

Description

[Technical Field]

[0001] The present invention relates to novel compounds that can degrade target proteins or polypeptides using ubiquitin, and more particularly to compounds that have a UBM (ubiquitin binding moiety) structure that can be covalently linked to a TBM (targeted protein binding moiety) structure containing a ligand that binds to a target protein, and to which ubiquitin can directly bind. [Background technology]

[0002] Ubiquitin is a molecular identifier that is covalently attached to the terminus of proteins requiring degradation. This process involves the in vivo enzymes E1 (ubiquitin-activating enzyme), E2 (ubiquitin carrier protein), and E3 (ubiquitin-protein ligase, E3 ligase). The attachment of ubiquitin to a protein is called ubiquitination. When multiple ubiquitins are attached to a protein (polyubiquitylation), the protein is degraded by the intracellular proteolytic mechanisms, including the proteasome and lysosome. Protein residues degraded in this way can be reused within the cell. The process of protein degradation by endosomes or autophagosomes, where proteins are recognized by endosomes or autophagosomes and then bound to lysosomes, is called endo / autolysosomal degradation. The process by which proteins are targeted to the proteasome for degradation is called the ubiquitin-proteasome system (UPS). The ubiquitination degradation pathway is particularly useful for targeting proteins or polypeptides, particularly when targeting kinases or specific receptors.

[0003] On the other hand, to utilize the ubiquitin-proteasome system, bifunctional compounds consisting of a moiety that binds to a target protein or polypeptide and a moiety that binds to an E3 enzyme are used, linked by a linker. This technology, which induces target protein degradation by ubiquitinating the lysine residues of the target protein, is called proteolysis-targeting chimera (PROTAC). PROTACs can degrade disease-related proteins that are difficult to control with existing small molecule compounds, antibodies, or drugs, such as transcription factors, scaffolding proteins, and tau proteins. They have the advantages of high target selectivity and reusability, allowing for low-dose administration. Furthermore, in diseases caused by ligand-receptor binding, neutralizing the action of excess ligands by blocking the ligand or using antagonists is a treatment with very low selectivity, with a high risk of patient resistance. Therefore, a more effective approach is to degrade the receptor protein itself that binds to the ligand.

[0004] However, the effectiveness of PROTAC compounds in degrading target proteins varies greatly depending on the type of E3 enzyme they bind to. Of the approximately 600 or so E3 enzymes, only about five are currently utilized in PROTAC technology, primarily CRBN, VHL, and IAP. Furthermore, PROTAC compounds, in which both the target protein-binding moiety and the E3 enzyme-binding moiety are peptides, have the disadvantage of low cell permeability. To address this issue, small molecules rather than peptides can be used as E3 enzyme-binding ligands. Some examples in WO 2020 / 251971 have reported that lysine-mimetic moieties can be ubiquitinated, enabling degradation via the ubiquitin-proteasome pathway (UPP). However, no commercial products have been developed yet, and research and development of related technologies is needed. Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide a compound having a UBM (ubiquitin binding moiety) structure to which ubiquitin can be directly bound.

[0006] Another object of the present invention is to provide a compound in which a compound having the UBM structure and a compound having a TBM (targeted protein / polypeptide binding moiety) structure are linked to each other. [Means for solving the problem]

[0007] To achieve the above object, one aspect of the present invention provides compounds represented by the following chemical formulas I to IV, or pharmaceutically acceptable salts, isomers, solvates, hydrates, or prodrugs thereof:

[0008] [Chemical formula I] TIFF2025503429000002.tif20170

[0009] [Chemical formula II] TIFF2025503429000003.tif20170

[0010] [Chemical formula III] TIFF2025503429000004.tif20170

[0011] [Chemical formula IV] TIFF2025503429000005.tif20170

[0012] In order to achieve the above object, in yet another aspect, the present invention provides a compound or a pharmaceutically acceptable salt, isomer, solvate, hydrate, or prodrug thereof, which has a chemical structure of TBM-UBM, wherein the TBM is a moiety that binds to a target protein or polypeptide, and the UBM has a chemical structure represented by any one of the following chemical formulas V to VIII:

[0013] [Chemical formula V] TIFF2025503429000006.tif20170

[0014] [Chemical formula VI] TIFF2025503429000007.tif20170

[0015] [Chemical formula VII] TIFF2025503429000008.tif20170

[0016] [Chemical formula VIII] TIFF2025503429000009.tif20170 [Effects of the Invention]

[0017] The compounds according to the present invention contain a UBM structure capable of inducing polyubiquitination of a target protein or polypeptide, and can function as degraders capable of selectively degrading the target protein or polypeptide via the proteasome or lysosome. That is, target proteins or polypeptides ubiquitinated by the UBM can be subjected to endo- or autolysosomal degradation in addition to targeted proteolysis using the UPS, as in conventional PROTACs. Therefore, the TBM-UBM compounds of the present invention have the advantage of being able to selectively target and degrade protein complexes and protein aggregates.

[0018] However, the effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the following description. [Brief explanation of the drawings]

[0019] [Figure 1]1 is a schematic diagram showing the principle of operation of a proteolyzer formed by binding of the TBM-UBM compound of the present invention to a target protein or polypeptide. [Figure 2] This shows the results of a pulldown assay carried out to confirm that the UBM compound is a receptor that directly binds to ubiquitin. [Figure 3] FIG. 1 shows mass spectrometry results from a prototype protein analysis method performed to confirm that BRD4 degrader directly binds to ubiquitin. DETAILED DESCRIPTION OF THE INVENTION

[0020] First, the terms used in the present invention will be defined.

[0021] For purposes of the present invention, the following terms have the following meanings unless otherwise specified: Any term not defined has its art-understood meaning.

[0022] Throughout this specification, unless specifically stated to the contrary, the term "comprising" an element means that it may further include other elements, rather than excluding other elements.

[0023] In the structural formulas herein, the symbol "-" connecting atoms and / or groups may mean a single bond, and the symbol "=" may mean a double bond. The symbols may be omitted or may be displayed when necessary, such as when specifying the bonded atom or bond position.

[0024] As used herein, "halogen" refers to fluoro (F), chlorine (Cl), bromine (Br), and iodine (I).

[0025] The term "alkyl" as used herein refers to a cyclic or acyclic aliphatic hydrocarbon group containing no double or triple bonds, and unless otherwise specified, the acyclic alkyl may have 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, or 1 to 10 carbon atoms, for example, 1 to 6 carbon atoms, particularly 1 to 4 carbon atoms, and may be linear or branched. Specific examples of the alkyl group include a methyl group, an ethyl group, a propyl group, an n-propyl group, an isopropyl group, a butyl group, an n-butyl group, an isobutyl group, a tert-butyl group, a sec-butyl group, a 1-methylbutyl group, a 1-ethylbutyl group, a pentyl group, an n-pentyl group, an isopentyl group, a neopentyl group, a tert-pentyl group, a hexyl group, an n-hexyl group, a 1-methylpentyl group, a 2-methylpentyl group, a 4-methyl-2-pentyl group, a 3,3-dimethyl- Examples of such alkyl groups include, but are not limited to, ethylbutyl, 2-ethylbutyl, heptyl, n-heptyl, 1-methylhexyl, octyl, n-octyl, tert-octyl, 1-methylheptyl, 2-ethylhexyl, 2-propylpentyl, n-nonyl, 2,2-dimethylheptyl, 1-ethylpropyl, 1,1-dimethylpropyl, isohexyl, 4-methylhexyl, 5-methylhexyl, and benzyl. Furthermore, in the present invention, the cyclic alkyl is particularly referred to as "cycloalkyl", and the "cycloalkyl" is a cyclic aliphatic hydrocarbon group containing no double or triple bonds, and unless otherwise specified, may have 3 to 30, 3 to 28, 3 to 26, 3 to 24, 3 to 22, 3 to 20, 3 to 18, 3 to 16, 3 to 14, 3 to 12, or 3 to 10 carbon atoms, for example, 3 to 8 carbon atoms, particularly 3 to 6 carbon atoms, and may be a monocyclic or polycyclic group. The polycyclic group refers to a group in which a cycloalkyl group is directly linked to or condensed with another cyclic group, and here, the other cyclic group may be a cycloalkyl group or may be another type of cyclic group, for example, a heterocycloalkyl group, an aryl group, or a heteroaryl group.Specific examples of the cycloalkyl group include, but are not limited to, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a 3-methylcyclopentyl group, a 2,3-dimethylcyclopentyl group, a cyclohexyl group, a 3-methylcyclohexyl group, a 4-methylcyclohexyl group, a 2,3-dimethylcyclohexyl group, a 3,4,5-trimethylcyclohexyl group, a 4-tert-butylcyclohexyl group, a cycloheptyl group, a cyclooctyl group, a cyclononyl group, a cyclodecyl group, a cycloundecyl group, a cyclododecyl group, a bicyclo[2.2.1]heptyl group, a bicyclo[2.2.2]octyl group, a bicyclo[3.2.2]nonyl group, a bicyclo[4.4.0]decyl group, and a bicyclo[4.1.0]heptyl group.

[0026] The term "alkenyl" as used herein refers to a cyclic or acyclic aliphatic hydrocarbon group containing at least one double bond, and unless otherwise specified, the acyclic alkenyl may have 2 to 20, 2 to 19, 2 to 18, 2 to 17, 2 to 16, 2 to 15, 2 to 14, 2 to 13, 2 to 12, 2 to 11, or 2 to 10 carbon atoms, for example, 2 to 6 carbon atoms, particularly 2 to 4 carbon atoms, and may be linear or branched. Specific examples of the alkenyl group include, but are not limited to, a vinyl group, a 1-propenyl group, an isopropenyl group, a 1-butenyl group, a 2-butenyl group, a 3-butenyl group, a 1-pentenyl group, a 2-pentenyl group, a 3-pentenyl group, a 3-methyl-1-butenyl group, a 1,3-butadienyl group, an allyl group, a 1-phenylvinyl-1-yl group, a 2-phenylvinyl-1-yl group, a 2,2-diphenylvinyl-1-yl group, a 2-phenyl-2-(naphthyl-1-yl)vinyl-1-yl group, a 2,2-bis(diphenyl-1-yl)vinyl-1-yl group, a stilbenyl group, and a styrenyl group. Furthermore, in the present invention, the cyclic alkenyl is particularly referred to as "cycloalkenyl," and the "cycloalkenyl" is a cyclic aliphatic hydrocarbon group containing at least one double bond, and unless otherwise specified, may have 3 to 30, 3 to 28, 3 to 26, 3 to 24, 3 to 22, 3 to 20, 3 to 18, 3 to 16, 3 to 14, 3 to 12, or 3 to 10 carbon atoms, for example, 3 to 8 carbon atoms, particularly 3 to 6 carbon atoms, and may be monocyclic or polycyclic. The polycyclic ring refers to a group in which a cycloalkenyl group is directly linked to or condensed with another cyclic group, and here, the other cyclic group may be a cycloalkyl group or may be another type of cyclic group, for example, a heterocycloalkyl group, an aryl group, or a heteroaryl group. Specific examples of the cycloalkenyl group include, but are not limited to, a cyclopentenyl group, a cyclohexenyl group, a cyclopenta-1,3-dienyl group, a cycloheptenyl group, a cyclooctenyl group, and a cycloocta-1,4-dienyl group.

[0027] The term "alkynyl" as used herein refers to an aliphatic hydrocarbon group containing at least one triple bond, and unless otherwise specified, may have 2 to 20, 2 to 19, 2 to 18, 2 to 17, 2 to 16, 2 to 15, 2 to 14, 2 to 13, 2 to 12, 2 to 11, or 2 to 10 carbon atoms, for example, 2 to 6 carbon atoms, particularly 2 to 4 carbon atoms, and may be linear or branched. Specific examples of the alkynyl group include, but are not limited to, an ethynyl group, a propynyl group, a butynyl group, a pentynyl group, and a hexynyl group.

[0028] The term "aryl" as used herein refers to an aromatic hydrocarbon group, and unless otherwise specified, may have 6 to 30, 6 to 28, 6 to 26, 6 to 24, 6 to 22, 6 to 20, 6 to 18, 6 to 16, or 6 to 14 carbon atoms, for example, 6 to 12 carbon atoms, and may be monocyclic or polycyclic. The polycyclic group refers to a group in which an aryl group is directly linked to or condensed with a different cyclic group, and here, the other cyclic group may be an aryl group, but may also be other types of cyclic groups, such as a cycloalkyl group, a heterocycloalkyl group, or a heteroaryl group. The aryl group also includes a spiro group. Specific examples of the aryl group include, but are not limited to, a phenyl group, a biphenyl group, a triphenyl group, a naphthyl group, an anthryl group, a chrysenyl group, a phenanthrenyl group, a perylenyl group, a fluoranthenyl group, a triphenylenyl group, a phenalenyl group, a pyrenyl group, a tetracenyl group, a pentacenyl group, a fluorenyl group, an indenyl group, an acenaphthylenyl group, a benzofluorenyl group, a spirobifluorenyl group, a 2,3-dihydro-1H-indenyl group, and fused ring groups thereof.

[0029] The terms "heterocycloalkyl," "heterocycloalkenyl," and "heteroaryl" used herein mean that at least one atom constituting the ring of the above-mentioned cycloalkyl, cycloalkenyl, and aryl is substituted with a heteroatom such as O, S, Se, N, or Si, respectively.

[0030] As used herein, "pharmaceutically acceptable" means capable of being approved by a federal or state regulatory agency, or preferably approved, or listed in the United States Pharmacopoeia, or recognized by other commonly used pharmacopeias, for use in animals, and more particularly, in humans, by avoiding substantial toxic effects when used in normal medicinal dosages.

[0031] As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention that is pharmaceutically acceptable and possesses the desired biological or pharmacological activity of the parent compound. Examples of such salts include, but are not limited to, acid addition salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.), as well as acid addition salts formed with acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, fumaric acid, maleic acid, ascorbic acid, trifluoroacetic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalene sulfonic acid, naphthalene disulfonic acid, and the like. These include salts formed with organic acids such as methyl methyl esters of phenylalanine, ...

[0032] As used herein, the term "hydrate" refers to a compound of the present invention or a salt thereof that contains a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces.

[0033] As used herein, the term "solvate" refers to a compound of the present invention or a salt thereof that contains a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. Preferred solvents in this regard are volatile, non-toxic, and / or suitable for administration to humans.

[0034] As used herein, the term "prodrug" refers to a substance that is transformed into a parent drug in vivo. This refers to a compound of the present invention that can be hydrolyzed, oxidized, or undergo other reactions under biological conditions (in vitro or in vivo) to provide an active compound, particularly a compound of the present invention. Examples of prodrugs include compounds that are biohydrolyzed to produce a compound of the present invention, including biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogs, but are not limited to these specific embodiments. Such prodrugs include compounds that are easily prepared based on various known literature.

[0035] As used herein, the term "isomer" refers to a compound of the present invention or a salt thereof that has the same chemical or molecular formula but is structurally or sterically different. Such isomers include structural isomers such as tautomers, stereoisomers such as R or S isomers having asymmetric carbon centers, geometric isomers (trans, cis), and enantiomers. All of these isomers and mixtures thereof are also included within the scope of the present invention.

[0036] As used herein, the term "pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient, or carrier with which a compound of the present invention is administered.

[0037] As used herein, "prevention" means reducing the risk of acquiring a disease or disorder (i.e., preventing one or more clinical symptoms of a disease from occurring in an individual who is exposed to or susceptible to developing the disease but who has not yet developed the disease or is asymptomatic of the disease).

[0038] As used herein, "treatment" means ameliorating a disease or disorder (i.e., arresting or reducing the progression of the disease or one or more clinical symptoms of the disease), or improving one or more physical parameters that are not discernible by an individual, or modulating a disease or disorder physically (e.g., stabilizing symptoms that are not discernible), or mentally (e.g., stabilizing physical parameters), or both.

[0039] The present invention will be described in detail below.

[0040] 1. UBM (ubiquitin binding moiety) compounds One aspect of the present invention provides compounds represented by the following chemical formulas I to IV, or pharmaceutically acceptable salts, isomers, solvates, hydrates or prodrugs thereof:

[0041] [Chemical formula I] TIFF2025503429000010.tif20170

[0042] [Chemical formula II] TIFF2025503429000011.tif20170

[0043] [Chemical formula III] TIFF2025503429000012.tif20170

[0044] [Chemical formula IV] TIFF2025503429000013.tif20170

[0045] In the formulas I to IV, the substituents W1, A, R1, R2, X1, X2, X3, B, W2′, Y1′, and Y2′ may each independently be defined as follows:

[0046] W1 may be any one selected from the group consisting of -alkylene-, -alkylene-O-, -alkylene-OSO2-, and -alkylene-N(R')SO2-.

[0047] W1 may be substituted or unsubstituted with at least one selected from the group consisting of deuterium, alkyl, C(O)OR', C(O)N(R')(R"), alkylene-C(O)OR', alkylene-C(O)N(R')(R"), alkylene-OR', alkylene-SR', alkylene-N(R')(R"), alkylene-OP(O)(OR')(OR"), alkylene-P(O)(OR')(OR"), and alkylene-OP(O)(R')(R")). For example, W1 may be substituted or unsubstituted with at least one selected from the group consisting of C(O)OR', C(O)N(R')(R"), alkylene-OR', alkylene-SR', and alkylene-OP(O)(OR')(OR"). Specifically, W1 may be substituted or unsubstituted with at least one selected from the group consisting of alkylene-OR' and alkylene-OP(O)(OR')(OR'').

[0048] A may be any one selected from the group consisting of aryl, heteroaryl, cycloalkyl, heterocycloalkyl, and heterocycloalkenyl. For example, A may be any one selected from the group consisting of aryl, 5- to 10-membered heteroaryl containing at least one selected from the group consisting of N, O, and S, 3- to 10-membered cycloalkyl, 3- to 10-membered heterocycloalkyl containing at least one selected from the group consisting of N, O, and S, and 3- to 10-membered heterocycloalkenyl containing at least one selected from the group consisting of N, O, and S. Specifically, A may be any one selected from the group consisting of benzene, furan, benzofuran, pyridine, thiophene, imidazole, and oxazole.

[0049] R1 and R2 each independently represent hydrogen, alkyl, alkenyl, alkynyl, halogen, oxo, CN, NO2, OR', SR', N(R')(R''), O-(alkylene)-OR', -N(R')-(alkylene)-OR', C(O)R', C(NH)R', C(S)R', C(O)OR', C(O)N(R')(R''), P(O)(R')(R''), P(O)(OR')(OR''), C(N H)N(R')(R''), C(S)OR', C(S)N(R')(R''), C(O)O-(alkylene)-OC(O)R', C(O)O-(alkylene)-methacryloyloxy, (alkylene)-OR', (alkylene)-SR', (alkylene)-N(R')(R''), (alkynylene)-N(R')(R''), (alkylene)-P(O)(R')(R'')-, (alkylene)-P(O)(OR') (OR''), (alkylene)-OP(O)(R')(R''), (alkylene)-OP(O)(OR')(OR''), (alkylene)-OP(O)(benzyloxy)2, (alkylene)-OP(O)(phenoxy)2, (alkylene)-OP(O)(phenoxy)(NH-CH(R')C(O)OR''), (alkylene)-C(O)OR', (alkenylene)-C(O)OR', (alkynylene-C(O )OR′, (alkylene)-C(O)N(R′)(R″), (alkenylene)-C(NH)N(R′)(R″), (alkynylene)-C(S)N(R′)(R″), (alkylene)-C(O)R′, (alkylene)-C(NH)R′, (alkylene)-C(S)R′, aryl, heteroaryl, heterocycloalkyl, and heterocycloalkenyl.For example, R1 and R2 are each independently hydrogen, alkyl, alkenyl, halogen, OR', N(R')(R''), O-(alkylene)-OR', C(O)R', C(O)OR', C(O)N(R')(R''), C(NH)N(R')(R''), C(O)O-(alkylene)-OC(O)R', C(O)O-(alkylene)-methacryloyloxy, (alkylene)-OR', (alkylene)-SR', (alkylene)-N(R')(R''), (a and R may be any one selected from the group consisting of (alkylene)-OP(O)(OR')(OR"), (alkylene)-OP(O)(benzyloxy), (alkylene)-OP(O)(phenoxy), (alkylene)-OP(O)(phenoxy)(NH-CH(R')C(O)OR"), (alkylene)-C(O)OR', aryl, heteroaryl, heterocycloalkyl, and heterocycloalkenyl. Specifically, R and R may each independently be any one selected from the group consisting of hydrogen, alkyl, OR', C(O)R', C(O)OR', C(O)N(R')(R"), (alkylene)-OR', (alkylene)-SR', (alkylene)-OP(O)(OR')(OR"), heteroaryl, and heterocycloalkyl.

[0050] Furthermore, R1 and R2 may each independently be substituted with one or more halogen atoms, and when R1 and R2 are aryl, heteroaryl, heterocycloalkyl, or heterocycloalkenyl, they may each independently be substituted at one or more positions with one or more halogen-substituted or unsubstituted alkyl or N(R')(R'').

[0051] X1 may be a direct bond or any one selected from the group X1a consisting of —N(R′)—, —O—, —S—, —C(O)—, —SO—, SO2—, =NO— and —ON=; the group X1b consisting of —OC(O)—, —C(O)O—, —N(R′)C(O)—, —C(O)N(R′)—, —N(R′)SO—, —SON(R′)—, —N(R′)SO2— and —SON(R′)—; and the group X1c consisting of -alkylene-, -alkenylene- and -alkynylene-, or two or more of them may be linked together. For example, X1 may be a direct bond, any one selected from the X1a group and the X1b group, or any one selected from the X1a group and the X1b group and any one selected from the X1c group linked in the form of X1a-X1c or X1b-X1c. Specifically, X1 may be a direct bond, or the X1a group constituting X1 may consist of -N(R')- and -O-, the X1b group may consist of -C(O)O-, -N(R')C(O)- and -C(O)N(R')-, or the X1c group may consist of -alkylene- and -alkynylene-.

[0052] X2 may be a direct bond or any one selected from the group X2a consisting of -N(R')-, -O-, -S-, -C(O)-, -SO-, SO2-, =NO- and -ON=; the group X2b consisting of -OC(O)-, -N(R')C(O)-, -N(R')C(O)O-, -N(R')S02-O- and -N(R')C(S)O-; the group X2c consisting of -C(O)O-, -C(O)N(R')-, -O-SO2-N(R')-, -OC(S)N(R')- and -OC(O)N(R')-; and the group X2d consisting of -alkylene-, -alkenylene- and -alkynylene-, or two or more of them may be bonded to each other. For example, X2 may be a direct bond, any one selected from the X2a group, the X2b group, and the X2c group, or two or more of them linked to each other, or any one or more selected from the X2b group, the X2c group, and the X2d group may be linked in the form of X2b-X2d-X2b, X2b-X2d-X2c, X2c-X2d-X2b, or X2c-X2d-X2c. Specifically, X2 may be a direct bond, or the X2a group constituting X2 may consist of -N(R')-, -O-, -C(O)-, and =NO-, the X2b group may consist of -N(R')C(O)-, the X2c group may consist of -C(O)N(R')-, or the X2d group may consist of alkylene-, alkenylene-, and -alkynylene-.

[0053] X3 may be a direct bond or any one selected from the group consisting of -alkylene-, -O-, -S-, -C(O)-, -N(R')-, -SO-, -SO2-, and combinations thereof. For example, X3 may be a direct bond or any one selected from the group consisting of -C(O)-, -N(R')-, and combinations thereof.

[0054] B may be any one selected from the group consisting of aryl, heteroaryl, cycloalkyl, heterocycloalkyl, and heterocycloalkenyl, each substituted or unsubstituted with at least one selected from the group consisting of hydroxy, alkoxy, and oxo, or two or more selected from the group consisting of aryl, heteroaryl, cycloalkyl, heterocycloalkyl, and heterocycloalkenyl, each substituted or unsubstituted with at least one selected from the group consisting of hydroxy, alkoxy, and oxo, linked by a direct bond, -alkylene-, -O-, -S-, -C(O)-, -N(R')-, -SO-, -SO2-, or a combination thereof. For example, B may be any one selected from the group consisting of aryl, 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, all of which may be substituted or unsubstituted with at least one selected from the group consisting of hydroxy, alkoxy, and oxo, or two or more selected from the group consisting of aryl, 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, all of which may be substituted or unsubstituted with at least one selected from the group consisting of hydroxy and oxo, linked directly by -alkylene-, -C(O)-, -N(R')-, or a combination thereof. Specifically, B is any one selected from the group consisting of 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, all of which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy and oxo, or two or more selected from the group consisting of unsubstituted 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, which are directly linked via -alkylene-, -C(O)-, -N(R')-, or a combination thereof.

[0055] W2' may be any one selected from the group consisting of hydrogen, alkyl, alkenyl, and alkynyl. For example, W2' may be any one selected from the group consisting of hydrogen, alkenyl, and alkynyl. Specifically, W2' may be any one selected from the group consisting of hydrogen and alkynyl.

[0056] In addition, W2' may be deuterium, halogen, alkyl, -alkylene-C(O)OR', alkylene-C(O)N(R')(R''), alkylene-OR', alkylene-SR', alkylene-N(R')(R''), alkylene-OP(O)(OR')(OR''), alkylene-P(O)(OR')(OR''), alkylene-OP(O)(R')(R''), -N(R')(R''), -OR', -SR', -C( W may be substituted or unsubstituted with at least one selected from the group consisting of: -O)R', -SOR', -SOR', =N-OR', -C(O)OR', -C(O)N(R')(R''), -OC(O)OR', -OC(O)N(R')(R''), -N(R')C(O)O(R''), -N(R')SO(R'), -SON(R')(R''), -N(R')SO2(R'') and -SON(R')(R''). For example, W may be substituted or unsubstituted with at least one selected from the group consisting of halogen, alkylene-OR', alkylene-OP(O)(OR')(OR''), -N(R')(R''), -OR' and -C(O)OR'.

[0057] The Y' is an alkyl, alkenyl, alkynyl, (CHCHO)-(CHCHO) n-1 CH2- and (CH3CH2O)-(CH2CH2O) n-1 CH2CH2-, and n may be 1 to 4. For example, Y1' is alkyl, alkenyl, alkynyl, (CH3CH2O)-(CH2CH2O) n-1 CH2- and (CH3CH2O)-(CH2CH2O) n-1CH2CH2-, and n may be 1 or 2. Specifically, Y1' may be any one selected from the group consisting of alkyl, alkenyl, and alkynyl.

[0058] Furthermore, Y1' may be unsubstituted or substituted with at least one selected from the group consisting of halogen, -N(R')(R''), -OR', -SR', -C(O)R', -SOR', SOR', =N-OR', -C(O)OR', -C(O)N(R')(R''), -OC(O)OR', -OC(O)N(R')(R''), -N(R')C(O)O(R''), -N(R')SO(R'), -SON(R')(R''), -N(R')SO2(R''), and -SON(R')(R''). Specifically, Y1' may be unsubstituted or substituted with at least one selected from the group consisting of halogen, -N(R')(R''), -OR', and -C(O)OR'.

[0059] Y2' may be any one selected from the group consisting of alkyl, alkenyl, alkynyl, heterocycloalkyl, and heterocycloalkenyl. For example, Y2' may be any one selected from the group consisting of alkyl, alkenyl, alkynyl, and heterocycloalkyl. Specifically, Y2' may be any one selected from the group consisting of alkyl and heterocycloalkyl.

[0060] Furthermore, Y2' may be unsubstituted or substituted with at least one selected from the group consisting of halogen, -N(R')(R''), -OR', -SR', -C(O)R', -SOR', SOR', =N-OR', -C(O)OR', -C(O)N(R')(R''), -OC(O)OR', -OC(O)N(R')(R''), -N(R')C(O)O(R''), -N(R')SO(R'), -SON(R')(R''), -N(R')SO2(R''), and -SON(R')(R''). For example, Y2' may be unsubstituted or substituted with at least one selected from the group consisting of halogen, -N(R')(R''), -OR', and -C(O)OR'.

[0061] On the other hand, in the functional group constituting the substituent, R' and R'' may each independently be hydrogen or alkyl.

[0062] In particular, specific examples of the compound represented by Formula I may be those listed in Table 1 below, but are not limited thereto.

[0063] [Table 1] TIFF2025503429000015.tif246170TIFF2025503429000016.tif242170TIFF2025503429000017.tif213170TIFF20255034290 00018.tif245170TIFF2025503429000019.tif244170TIFF2025503429000020.tif241170TIFF2025503429000021.tif235170 TIFF2025503429000022.tif242170TIFF2025503429000023.tif239170TIFF2025503429000024.tif245170TIFF20255034290 00025.tif247170TIFF2025503429000026.tif225170TIFF2025503429000027.tif244170TIFF2025503429000028.tif108170

[0064] In particular, specific examples of the compound represented by Formula II may be those listed in Table 2 below, but are not limited thereto.

[0065] [Table 2] TIFF2025503429000030.tif232170TIFF2025503429000031.tif231170TIFF2025503429000032.tif241170TIFF20255034290000 33.tif239170TIFF2025503429000034.tif243170TIFF2025503429000035.tif211170TIFF2025503429000036.tif238170TIFF202 5503429000037.tif241170TIFF2025503429000038.tif240170TIFF2025503429000039.tif242170TIFF2025503429000040.tif23 4170TIFF2025503429000041.tif230170TIFF2025503429000042.tif238170TIFF2025503429000043.tif234170TIFF20255034290 00044.tif223170TIFF2025503429000045.tif233170TIFF2025503429000046.tif241170TIFF2025503429000047.tif241170TIF F2025503429000048.tif240170TIFF2025503429000049.tif231170TIFF2025503429000050.tif224170TIFF2025503429000051.t if240170TIFF2025503429000052.tif245170TIFF2025503429000053.tif242170TIFF2025503429000054.tif228170TIFF2025503 429000055.tif244170TIFF2025503429000056.tif228170TIFF2025503429000057.tif236170TIFF2025503429000058.tif234170

[0066] In particular, specific examples of the compound represented by Formula III may be, but are not limited to, those listed in Table 3 below.

[0067] [Table 3] TIFF2025503429000060.tif233170TIFF2025503429000061.tif241170TIFF2025503429000062.tif237170TIFF202 5503429000063.tif210170TIFF2025503429000064.tif245170TIFF2025503429000065.tif242170TIFF20255034290 00066.tif235170TIFF2025503429000067.tif241170TIFF2025503429000068.tif232170TIFF2025503429000069.t if243170TIFF2025503429000070.tif209170TIFF2025503429000071.tif212170TIFF2025503429000072.tif153170

[0068] In particular, specific examples of the compound represented by Formula IV may be those listed in Table 4 below, but are not limited thereto.

[0069] [Table 4] TIFF2025503429000074.tif245170TIFF2025503429000075.tif245170TIFF2025503429000076.tif219170TIFF2025503429000077.tif210170

[0070] In particular, in the compounds represented by Chemical Formulae I to IV, the amine group (-NH2) present in the propargyl group containing W1 is the position to which ubiquitin binds. Therefore, the compounds represented by Chemical Formulae I to IV can act as degraders of other target proteins or polypeptides by themselves or by binding to a ligand that specifically binds to the target protein or polypeptide. As described above, in the compounds represented by Chemical Formulae I to IV, the amine group (-NH2) present in the propargyl group containing W1 is the position to which ubiquitin binds. When the compounds represented by Chemical Formulae I to IV are linked to a ligand for the other target protein or polypeptide, it is preferable that the ligand be linked to a position other than the amine group (-NH2) present in the propargyl group containing W1.

[0071] 2. target protein / polypeptide degrader Another aspect of the present invention provides a compound having the chemical structure of TBM-UBM or a pharmaceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof.

[0072] The UBM may have a chemical structure represented by the following chemical formulas V to VIII.

[0073] [Chemical formula V] TIFF2025503429000078.tif20170

[0074] [Chemical formula VI] TIFF2025503429000079.tif20170

[0075] [Chemical formula VII] TIFF2025503429000080.tif20170

[0076] [Chemical formula VIII] TIFF2025503429000081.tif20170

[0077] In the formulae V to VIII, the substituents W1, A, R1, R2, X1, X2, X3, and B are each independently as described in connection with the compounds represented by the formulae I to IV.

[0078] Meanwhile, in the formulae V to VIII, the substituents W2, Y1 and Y2 can be independently defined as follows:

[0079] W2 may be a direct bond or any one selected from the group consisting of -alkylene-, -alkenylene-, and -alkynylene-. For example, W2 may be a direct bond or any one selected from the group consisting of -alkenylene- and -alkynylene-. Specifically, W2 may be a direct bond or -alkynylene-.

[0080] Furthermore, W2 may be substituted or unsubstituted with at least one selected from the group consisting of deuterium, alkyl, C(O)OR', C(O)N(R')(R"), alkylene-C(O)OR', alkylene-C(O)N(R')(R"), alkylene-OR', alkylene-SR', alkylene-N(R')(R"), alkylene-OP(O)(OR')(OR"), alkylene-P(O)(OR')(OR"), and alkylene-OP(O)(R')(R")). For example, W2 may be substituted or unsubstituted with at least one selected from the group consisting of alkylene-OR', alkylene-SR', and alkylene-OP(O)(OR')(OR"). Specifically, W2 may be substituted or unsubstituted with at least one selected from the group consisting of alkylene-OR' and alkylene-OP(O)(OR')(OR'').

[0081] The Y1 is a direct bond, or an alkylene, alkenylene, alkynylene, or —(CH2CH2O) n CH2- and -(CH2CH2O) nand n may be 1 to 4. For example, Y1 is a direct bond, or an alkylene, alkenylene, alkynylene, or -(CH2CH2O) n CH2- and -(CH2CH2O) n CH2CH2-, and n may be 1 to 2. Specifically, Y1 may be any one selected from the group consisting of a direct bond, alkylene, alkenylene, and alkynylene.

[0082] Y2 may be any one selected from the group consisting of a direct bond, alkylene, alkenylene, alkynylene, heterocycloalkylene, and heterocycloalkenylene. For example, Y2 may be any one selected from the group consisting of a direct bond, alkylene, alkenylene, alkynylene, and heterocycloalkylene. Specifically, Y2 may be any one selected from the group consisting of a direct bond, alkylene, and heterocycloalkylene.

[0083] In addition, in the functional group constituting the substituent, R' and R'' may each independently be hydrogen or alkyl.

[0084] The TBM may be a moiety that binds to a target protein or polypeptide.

[0085] The target protein or polypeptide refers to a protein or polypeptide that is targeted by the compound of the present invention for binding and degradation. The target protein or polypeptide includes any protein or polypeptide, as well as fragments, analogs, and / or homologs thereof. The target protein or polypeptide may be a protein or polypeptide with any biological function or activity, including structural, regulatory, hormonal, enzymatic, genetic, immunological, contractile, storage, transport, and signal transduction. By way of example, the target protein or polypeptide may be a structural protein, a receptor, an enzyme, a cell surface protein, a protein involved in catalytic activity, aromatase activity, motility activity, helicase activity, metabolic processes (anabolic and degradative metabolism), antioxidant activity, proteolysis, biosynthesis, a protein with kinase activity, oxidoreductase activity, transferase activity, hydrolase activity, lyase activity, isomerase activity, ligase activity, enzyme regulator activity, signal transducer activity, structural molecule activity, binding activity (proteins, lipids, carbohydrates), receptor activity, cell motility, membrane fusion, cell communication, regulation of biological processes, development, cell differentiation, response to stimuli, as well as proteins involved in the integrated function of cells, behavioral proteins, cell adhesion proteins, proteins involved in cell death, transport (including protein transporter activity, nuclear transport, ion transporter activity, pathway transporter activity, carrier activity, permease activity, secretion activity, electron transporter activity), pathogenesis, chaperone regulator activity, nucleic acid binding activity, transcription regulator activity, extracellular organization and biogenesis activity, translation regulator activity. In other examples, the target protein or polypeptide may be a protein or polypeptide derived from eukaryotes or prokaryotes, including humans, microorganisms, viruses, microorganisms, fungi and parasites, livestock and other animals, as well as microorganisms and other antagonistic microorganisms and plants for the determination of targets for antibiotics, among a variety of different targets for drug therapy.

[0086] More specifically, the target protein or polypeptide may be the target of many drugs used as therapeutic agents for humans. For example, the target protein or polypeptide may be a protein involved in cell death mechanisms, including B7.1, B7, TINFRlm, TNFR2, NADPH oxidase, and Bcl2Bax; a protein involved in the MAP / ERK signaling pathway, including Ras and Raf; a protein involved in the NF-κB signaling pathway; a protein involved in the mTOR signaling pathway; a protein involved in the wnt signaling pathway; a protein involved in the AKT signaling pathway; a protein involved in the JAK-STAT signaling pathway; a protein involved in the NOX complex and its signaling pathway; a protein involved in vascular endothelial growth factor and receptors and their signaling pathways; a protein involved in TLR receptors and their signaling pathways; a protein involved in G protein-coupled receptors and their signaling pathways; a protein involved in TCR-MHC complex-associated receptors and their signaling pathways; a protein involved in cell cycle checkpoint proteins and their signaling pathways; a protein including a factor regulating gene transcription and translation; a protein involved in gene replication and its signaling pathways; Regulators of, proteins including histone protein regulators for gene expression, factors regulating epigenetics and related proteins, proteins that constitute and regulate the cytoskeleton, proteins that regulate intracellular transport and proteins that regulate it, C5a receptor, dopamine receptor, 5HT receptor, chemokine receptor, histamine receptor, CD40 / CD40L, CD23, CD124, CD4, CD5, VCAM, VLA-4 integrin, cell lectins, ICOS, ICAM1, IL-2 receptor, IL-1 receptor, TN F-alpha receptors and other related cytokine and chemokine receptors, oxytocin receptors, glycine receptors, noradrenaline reuptake receptors, endothelin receptors, estrogen receptors, androgen receptors, adenosine receptors, TrkA receptors for NGF, vitronectin receptors, integrin receptors, neurokinins and receptors, epidermal growth factor receptors, purinergic receptors (P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7), EGF receptors, HMG-coA reductase, PDEPhosphodiesterase type V, PDE IV, phosphodiesterase type 4, PDE I, PDE II, PDE III, nitric oxide (NO) synthase, cyclooxygenase 1, cyclooxygenase 2, 5-lipoxygenase, trypsinase, serine proteases, thymidylate synthase, purine nucleoside phosphorylase, trypanosomal GAPDH, glycogen phosphorylase, carbonic anhydrase and its analogs, P-glycoprotein and MRP, tyrosine kinase, p56, lck, Cat+ pathway, inosine monophosphate dehydrogenase, interleukin-1 convertase, caspase-1, glycinamide ribonucleoside formyltransferase, poly(ADP-ribose) polymerase, acetylcholinesterase, cyclin-dependent kinase, microsomal transfer protein inhibitor, bile acid transport inhibitor, 5-alpha reductase inhibitor , angiotensin 11, adenosine receptor, adenosine kinase and AMP deaminase, farnesyltransferase, geranylgeranyltransferase, beta-amyloid, tyrosine kinase Her-21 / neu, telomerase inhibition, cytosolic phospholipase A2 and tyrosine kinase, acetyl-coA carboxylase, adenylosuccinate synthase, protoporphyrinogen oxidase and enolpyruvylshikimate-phosphate synthase, ecdysone 20-monooxygenase, GABA-gated chloride pathway, voltage-sensitive sodium pathway protein, sodium pathway, calcium release pathway protein, chloride pathway protein and polypeptides derived from this protein, HCV NS3 protease, HCV NS3 RNA helicase, rhinovirus 3C protease, herpes simplex virus-1 (HSV-I) protease, cytomegalovirus (CMV) protease, multidrug resistance (MDR) protein, HIV-1 protease, HIVThe target proteins or polypeptides include bacterial or viral proteins and polypeptides derived therefrom, such as integrase, influenza A, neuraminidase, and hepatitis B reverse transcriptase, as well as proteins and polypeptides such as vasoactive intestinal peptide, prion, neuropeptide Y, and receptors. In particular, the target protein or polypeptide may be a bromodomain-containing protein, Aurora kinase A, or p38 MAPK (mitogen activated protein kinase).

[0087] As described above, the TBM is a moiety that binds to the target protein or polypeptide, and various compounds known to bind to the target protein or polypeptide can be used as the TBM without limitation. For example, the TBM can include, but is not limited to, compounds targeting bromodomain-containing proteins, compounds targeting Aurora kinase A, and compounds targeting p38 MAPK. In particular, the compound targeting bromodomain-containing proteins can be a BRD4 inhibitor, which can have the chemical structure of Formula 1 below. Alternatively, the compound targeting Aurora kinase A can be an Aurora A inhibitor, which can have the chemical structure of Formula 2 below. Alternatively, the compound targeting p38 MAPK can be a p38 inhibitor, which can have the chemical structure of Formula 3 below.

[0088] [Chemical formula 1] TIFF2025503429000082.tif39170

[0089] [Chemical formula 2] TIFF2025503429000083.tif50170

[0090] [Chemical formula 3] TIFF2025503429000084.tif45170

[0091] Meanwhile, the TBM and UBM are linked by a covalent bond, and the covalent bond site may be a direct bond, or the TBM and UBM may each be modified to form any one selected from the group consisting of -N(R')-, -O-, -C(R')(R'')-O-, -S-, -C(O)-, -SO-, SO2-, =NO-, -ON=, -OC(O)-, -C(O)O-, -N(R')C(O)-, -C(O)N(R')-, -OC(O)O-, -OC(O)N(R')-, -N(R')C(O)O-, -N(R')SO-, -SON(R')-, -N(R')SO2- and -SON(R')-. For example, the TBM and UBM may each be modified so that the covalent bond site connecting the TBM and UBM is a direct bond or any one selected from the group consisting of -N(R')-, -O-, -C(R')(R'')-O-, -OC(O)-, -OC(O)N(R')-, and -C(O)N(R')-. Specifically, the TBM and UBM may each be modified so that the covalent bond site connecting the TBM and UBM is a direct bond or any one selected from the group consisting of -O-, -C(R')(R'')-O-, -OC(O)-, -OC(O)N(R')-, and -C(O)N(R')-.Therefore, for example, when the TBM is a compound having the chemical structure of Chemical Formula 1, it can be bonded to the UBM to form a compound having the chemical structure of Chemical Formula 4 below; when the TBM is a compound having the chemical structure of Chemical Formula 2, it can be bonded to the UBM to form a compound having the chemical structure of Chemical Formula 5 below; when the TBM is a compound having the chemical structure of Chemical Formula 3, it can be bonded to the UBM to form a compound having the chemical structure of Chemical Formula 6 below; and in Chemical Formulas 4 to 6 below, Z can be independently selected from the group consisting of: or any one selected from the group consisting of —N(R′)—, —O—, —C(R′)(R″)—O—, —S—, —C(O)—, —SO—, SO—, ═NO—, —ON=, —OC(O)—, —C(O)O—, —N(R′)C(O)—, —C(O)N(R′)—, —OC(O)O—, —OC(O)N(R′)—, —N(R′)C(O)O—, —N(R′)SO—, —SON(R′)—, —N(R′)SO— and —SON(R′)—, wherein R′ and R″ are each independently hydrogen or alkyl.

[0092] [Chemical formula 4] TIFF2025503429000085.tif37170

[0093] [Chemical formula 5] TIFF2025503429000086.tif45170

[0094] [Chemical formula 6] TIFF2025503429000087.tif48170

[0095] More specifically, in the compound represented by Chemical Formula V, specific examples of the compound of Chemical Formula 1 bonded as the TBM may be those shown in Table 5 below, specific examples of the compound of Chemical Formula 2 bonded as the TBM may be those shown in Table 6 below, and specific examples of the compound of Chemical Formula 3 bonded as the TBM may be those shown in Table 7 below, but are not limited thereto.

[0096]

Table 5

[0097]

Table 6

[0098] )

Table 7

[0099] More specifically, in the compound represented by Chemical Formula VI, specific examples of the compound represented by Chemical Formula 1 bonded as the TBM may be those shown in Table 8 below, specific examples of the compound represented by Chemical Formula 2 bonded as the TBM may be those shown in Table 9 below, and specific examples of the compound represented by Chemical Formula 3 bonded as the TBM may be those shown in Table 10 below, but are not limited thereto.

[0100] [Table 8] TIFF2025503429000117.tif246170TIFF2025503429000118.tif196170TIFF2025503429000119.tif239170TIFF2025503429000120.tif195170TIFF2025503429000121.tif245170TIFF2025503429000122.tif239170TIFF2025503429000123.tif242170TIFF2025503429000124.tif243170TIFF2025503429000125.tif236170TIFF2025503429000126.tif207170TIFF2025503429000127.tif234170TIFF2025503429000128.tif219170TIFF2025503429000129.tif200170TIFF2025503429000130.tif235170TIFF2025503429000131.tif234170TIFF2025503429000132.tif239170TIFF2025503429000133.tif219170TIFF2025503429000134.tif226170TIFF2025503429000135.tif209170TIFF2025503429000136.tif230170TIFF2025503429000137.tif246170TIFF2025503429000138.tif231170TIFF2025503429000139.tif220170TIFF2025503429000140.tif215170TIFF2025503429000141.tif218170TIFF2025503429000142.tif232170TIFF2025503429000143.tif234170TIFF2025503429000144.tif219170TIFF2025503429000145.tif215170TIFF2025503429000146.tif245170TIFF2025503429000147.tif231170TIFF2025503429000148.tif238170TIFF2025503429000149.tif223170TIFF2025503429000150.tif226170TIFF2025503429000151.tif215170TIFF2025503429000152.tif243170TIFF2025503429000153.ti f218170TIFF2025503429000154.tif220170TIFF2025503429000155.tif218170TIFF2025503429000156.tif234170TIFF2025503429000157.tif22 0170TIFF2025503429000158.tif231170TIFF2025503429000159.tif238170TIFF2025503429000160.tif183170TIFF2025503429000161.tif2381 70TIFF2025503429000162.tif213170TIFF2025503429000163.tif242170TIFF2025503429000164.tif231170TIFF2025503429000165.tif225170.

[0101] [Table 9] TIFF2025503429000167.tif225170TIFF2025503429000168.tif238170

[0102] [Table 10]

[0103] More specifically, in the compound represented by Chemical Formula VII, specific examples of the compound of Chemical Formula 1 bonded as the TBM may be those shown in Table 11 below, specific examples of the compound of Chemical Formula 2 bonded as the TBM may be those shown in Table 12 below, and specific examples of the compound of Chemical Formula 3 bonded as the TBM may be those shown in Table 13 below, but are not limited thereto.

[0104]

Table 11

[0105]

Table 12

[0106]

Table 13

[0107] More specifically, in the compound represented by Chemical Formula VIII, specific examples of the compound represented by Chemical Formula 1 bonded as the TBM may be those shown in Table 14 below, and specific examples of the compound represented by Chemical Formula 2 bonded as the TBM may be those shown in Table 15 below, but are not limited thereto.

[0108] [Table 14] TIFF2025503429000198.tif240170TIFF2025503429000199.tif215170TIFF2025503429000200.tif234170TIFF20255034290 00201.tif236170TIFF2025503429000202.tif237170TIFF2025503429000203.tif194170TIFF2025503429000204.tif189170

[0109] [Table 15]

[0110] As described above, the compounds of the present invention having the chemical structure of TBM-UBM can degrade target proteins or polypeptides. Specifically, as shown in Figure 1, after the TBM of the compounds of the present invention binds to a target protein or polypeptide, ubiquitin binds to the UBM of the compounds of the present invention ((poly)ubiquitination), and the target protein or polypeptide can be degraded by the proteasome or lysosome.

[0111] 3. Pharmaceutical Compositions Another aspect of the present invention provides a pharmaceutical composition comprising, as an active ingredient, a compound represented by any one of Formulas I to VIII, or a pharmaceutically acceptable salt, isomer, solvate, hydrate, or prodrug thereof.

[0112] As described above, the compounds represented by Formulas I to IV and the compounds represented by Formulas V to VIII have activity as degraders of target proteins or polypeptides, and therefore the pharmaceutical compositions of the present invention can be used for the prevention or treatment of diseases caused by such target proteins or polypeptides.

[0113] In particular, the pharmaceutical compositions of the present invention may contain, as an active ingredient, a compound represented by any one of Formulas V to VIII, or a pharmaceutically acceptable salt, isomer, solvate, hydrate, or prodrug thereof, wherein the specific use of the pharmaceutical composition may vary depending on the type of target protein or polypeptide to which the TBM binds.

[0114] For example, when the target protein or polypeptide to which the TBM binds is a bromodomain-containing protein such as BRD2, BRD3, BRD4, or BRDT, or when the TBM is a compound that targets a bromodomain-containing protein, such as a BRD4 inhibitor, the pharmaceutical composition can be used for the prevention or treatment of a bromodomain-containing protein-associated disease.

[0115] The bromodomain-containing protein-associated disease may be, for example, a disease or condition that develops, progresses, or worsens due to overexpression or overactivation of a bromodomain-containing protein such as BRD2, BRD3, BRD4, or BRDT, and specifically may be an acute or chronic autoimmune or inflammatory disease.

[0116] In yet another embodiment of the present invention, the bromodomain-containing protein-associated disease or condition is a disease caused by congenital or acquired hyperinflammatory response and persistent inflammation, such as acquired hemophilia A, alopecia areata, ankylosing spondylitis, anti-NMDA receptor encephalitis, catastrophic antiphospholipid syndrome (CAPS), Addison's disease, autoimmune autonomic ganglionopathy (AAG), autoimmune gastrointestinal motility disorder (AGDM), and others. dysmotility;AGID), autoimmune encephalitis, acute disseminated encephalomyelitis (ADEM), autoimmune gastritis, autoimmune hemolytic anemia (AIHA), autoimmune hepatitis (AIH), autoimmune hyperlipidemia, lymphocytic hypophysitis, autoimmune inner ear disease (AIED), autoimmune lymphoproliferative syndrome (ALPS), autoimmune myelofibrosis, autoimmune myocarditis, autoimmune oophoritis, autoimmune pancreatitis (AIP), autoimmune polyglandular syndrome type 1, 2, 3 (APS type 1, APS type 2, APS type 3, APECED), autoimmune progesterone dermatitis, autoimmune retinopathy (AIR), autoimmune sensory-acoustic hearing loss (SNHL), Barlow's syndrome, Behçet's disease, shotcretinopathy, choroidal sclerosis (BSCR), bullous pemphigoid, Castleman's disease, celiac disease, Chagas' disease, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic urticaria (CU), eosinophilic granulomatosis with polyangiitis (EGPA), Cogan's syndrome, cold agglutinin disease, CREST syndrome, Crohn's disease (CD), Cronkhite-Canada syndrome (CSS), cryptogenic organizing pneumoniapneumonia, COP), dermatomyositis, neuromyelitis optica (NMO), type 1 diabetes, Dressler syndrome, atopic dermatitis, endometriosis, eosinophilic esophagitis, eosinophilic fasciitis, erythema nodosum, primary macroglobulinemia, Evans syndrome, fibrinous pneumonia, giant cell arteritis, glomerulonephritis, Goodpasture syndrome, granulomatosis with polyangiitis (GPA), Graves' disease, Guillain-Barré syndrome (GBS), chronic lymphocytic thyroiditis, Henoch-Scholein purple Plaque disease, hidradenitis suppurativa, hypogammaglobulinemia, immunoglobulin A nephropathy, immune-mediated necrotizing myopathy (IMNM), immune thrombocytopenia (ITP), inclusion body myositis, IgG4-associated multiple sclerosis (ISD), interstitial cystitis, juvenile idiopathic arthritis, juvenile polymyositis, juvenile dermatomyositis, Kawasaki disease, Lambert-Eaton myasthenic syndrome (LEMS), leukocytoclastic vasculitis, lichen planus, lichen sclerosus, conjunctivitis, linear immunoglobulin A dermatosis (LAD), linear immunoglobulin A bullous dermatosis ( LABD), lupus nephritis, lymphocytic colitis, Meniere's disease, microscopic polyangiitis (MPA), mixed connective tissue disease (MCTD), Mooren's corneal ulcer, acute pityriasis lichenoides, multifocal motor neuropathy, multiple sclerosis (MS), myalgic encephalomyelitis (ME), myasthenia gravis (MG), ocular mucous membrane pemphigoid, opsoclonus-myoclonus syndrome (OMS), relapsing rheumatoid arthritis, paraneoplastic cerebellar degeneration, paraneoplastic pemphigus, progressive facial hemistrophy (HFA), paroxysmal nocturnal syndrome Hemoglobinuria, periphlebitis, Parsonage-Turner syndrome, pemphigoid of pregnancy, pemphigus foliaceus, pemphigus vulgaris, pernicious anemia, POEMS syndrome, polyarteritis nodosa (PAN), polymyalgia rheumatica, polymyositis, postural orthostatic tachycardia syndrome (POT), primary biliary cirrhosis (PBC), psoriasis, palmoplantar pustulosis, psoriatic arthritis, idiopathic pulmonary fibrosis (IPF), pure red cell aplasia (PRCA), pyoderma gangrenosum, Raynaud's disease, reactive arthritis, complex regional pain syndromesyndrome), relapsing polychondritis, restless legs syndrome (RLS), rheumatic fever, rheumatoid arthritis, sarcoidosis, autoimmune polyendocrine syndrome type 2, scleritis, scleroderma, serpiginous choroiditis, Sjogren's syndrome, stiff-person syndrome (SPS), small fiber neuropathy, systemic lupus erythematosus (SLE), Susac syndrome, Sydenham chorea, sympathetic fiber neuropathy, Takayasu's arteritis, orchitis, Tolosa-Hunt syndrome, transverse myelitis (TM), tubulointerstitial nephritis-uveitis syndrome (TINU), ulcerative colitis (UC), undifferentiated connective tissue disease, uveitis, vasculitis, leukoplakia, and Vogt-Koyanagi-Harada disease, and may also be applied to arteriosclerosis caused by the above diseases, and may be comprehensively applied to inflammatory diseases occurring in a subject.

[0117] In yet another embodiment of the present invention, the bromodomain-containing protein-associated disease or condition may be a cell proliferative disorder. Such cell proliferative disorders include, but are not limited to, cancerous hyperproliferative disorders (e.g., brain, lung, squamous cell, bladder, stomach, pancreas, breast, head, neck, kidney, liver, renal, ovarian, prostate, colorectal, colon, epidermal, esophageal, testicular, gynecological, or thyroid cancer, acute myeloid leukemia, multiple myeloma, mesothelioma, non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), neuroblastoma, and acute lymphocytic leukemia (ALL)); non-cancerous hyperproliferative disorders (e.g., benign hyperplasia of the skin (e.g., psoriasis), restenosis, and benign prostatic hyperplasia (BPH)); and angiogenesis or neovascularization-related diseases (e.g., tumor neovascularization, hemangioma, glioma, melanoma, Kaposi's sarcoma, and ovarian, breast, lung, pancreatic, prostate, colon, and epidermal carcinoma). Cell proliferative disorders further include primary and metastatic cancers.In particular, the compound is effective in the treatment of pseudomyxoma peritonei, intrahepatic and biliary tract cancer, hepatoblastoma, liver cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, oropharyngeal-tonsillar cancer (HPV-related), oropharyngeal-hypopharyngeal cancer (HPV-unrelated), mycosis fungoides, basal cell carcinoma, epithelial ovarian cancer, ovarian germ cell tumor, male breast cancer, brain tumor, pituitary adenoma, gallbladder and biliary tract cancer, gallbladder cancer, biliary tract cancer, colon cancer, chronic bone marrow cancer, and ovarian cancer. Myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, diffuse large B-cell lymphoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, sinus cancer, non-Hodgkin's lymphoma, tongue cancer, astrocytoma, childhood brain tumor, childhood lymphoma, childhood leukemia, small intestine cancer, meningioma, esophageal cancer, neuroblastoma, renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue tumor The compounds are useful in treating cancer in a subject, including, but not limited to, tumors, malignant bone tumors, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye tumors, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, gastric carcinoid, gastrointestinal stromal tumor, Wilms' tumor, breast cancer, sarcoma, penile cancer, gestational trophoblastic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone tumors, metastatic brain tumors, mediastinal cancer, rectal cancer, neuroendocrine tumors, colon cancer, vaginal cancer, spinal cord tumors, acoustic neuroma, pancreatic cancer, salivary gland tumors, Paget's disease, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, and thymic carcinoma, as well as metastatic disease and recurrence of the disease.

[0118] For example, when the target protein or polypeptide bound by the TBM is Aurora kinase A, or when the TBM is a compound that targets Aurora kinase A, such as an Aurora A inhibitor, the pharmaceutical composition can be used for the prevention or treatment of a cell proliferative disorder, such as those described above.

[0119] For example, when the target protein or polypeptide to which the TBM binds is the p38 MAPK, or when the TBM is a compound that targets p38 MAPK, such as a p38 inhibitor, the pharmaceutical composition can be used for the prevention or treatment of inflammation and inflammation-related diseases.

[0120] The inflammation or inflammation-related disease includes diseases caused by congenital or acquired hyperinflammatory reactions and persistent inflammation, such as acquired hemophilia A, alopecia areata, ankylosing spondylitis, anti-NMDA receptor encephalitis, antiphospholipid syndrome (CAPS), Addison's disease, autoimmune autonomic ganglionopathy (AAG), and autoimmune gastrointestinal motility disorder. dysmotility;AGID), autoimmune encephalitis, acute disseminated encephalomyelitis (ADEM), autoimmune gastritis, autoimmune hemolytic anemia (AIHA), autoimmune hepatitis (AIH), autoimmune hyperlipidemia, lymphocytic hypophysitis, autoimmune inner ear disease (AIED), autoimmune lymphoproliferative syndrome (ALPS), autoimmune myelofibrosis, autoimmune myocarditis, autoimmune oophoritis, autoimmune pancreatitis (AIP), autoimmune polyglandular syndrome type 1, 2, 3 (APS type 1, APS type 2, APS type 3, APECED), autoimmune progesterone dermatitis, autoimmune retinopathy (AIR), autoimmune sensory-acoustic hearing loss (SNHL), Barlow's syndrome, Behçet's disease, shotcretinopathy, choroidal sclerosis (BSCR), bullous pemphigoid, Castleman's disease, celiac disease, Chagas' disease, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic urticaria (CU), eosinophilic granulomatosis with polyangiitis (EGPA), Cogan's syndrome, cold agglutinin disease, CREST syndrome, Crohn's disease (CD), Cronkhite-Canada syndrome (CSS), idiopathic organizing pneumonia (COP), dermatomyositis, neuromyelitis optica (NMO), type 1 diabetes, Dressler's syndrome, atopic dermatitis, endometriosis, eosinophilic Esophagitis, eosinophilic fasciitis, erythema nodosum, primary macroglobulinemia, Evans syndrome, fibrinous pneumonia, giant cell arteritis, glomerulonephritis, Goodpasture's syndrome, granulomatosis with polyangiitis (GPA), Graves' disease, Guillain-Barré syndrome (GBS), chronic lymphocytic thyroiditis, Henoch-Scholein purpura, hidradenitis suppurativa, hypogammaglobulinemia, immunoglobulin A nephropathy, immune-mediated necrotizing myopathy (IMNM), immune thrombocytopenia (ITP), inclusion body myositis, IgG4-related multiple sclerosis (ISD), interstitial cystitis, juvenile idiopathic arthritis, juvenile polymyositis, juvenile dermatomyositis, Kawasaki disease, Lambert-Eaton myasthenic syndrome (LEMS),Leukocytoclastic vasculitis, lichen planus, lichen sclerosus, conjunctivitis, linear immunoglobulin A dermatosis (LAD), linear immunoglobulin A bullous dermatosis (LABD), lupus nephritis, lymphocytic colitis, Meniere's disease, microscopic polyangiitis (MPA), mixed connective tissue disease (MCTD), Mooren's corneal ulcer, acute pityriasis lichenoides, multifocal motor neuropathy, multiple sclerosis (MS), myalgic encephalomyelitis (ME), myasthenia gravis (MG), ocular mucous membrane pemphigoid, opsoclonus Myoclonus syndrome (OMS), relapsing rheumatism, paraneoplastic cerebellar degeneration, paraneoplastic pemphigus, progressive facial hemiatrophy (HFA), paroxysmal nocturnal hemoglobinuria, periphlebitis, Parsonage-Turner syndrome, pemphigoid of pregnancy, pemphigus foliaceus, pemphigus vulgaris, pernicious anemia, POEMS syndrome, polyarteritis nodosa (PAN), polymyalgia rheumatica, polymyositis, postural orthostatic tachycardia syndrome (POT), primary biliary cirrhosis (PBC) , psoriasis, palmoplantar pustulosis, psoriatic arthritis, idiopathic pulmonary fibrosis (IPF), pure red cell aplasia (PRCA), pyoderma gangrenosum, Raynaud's disease, reactive arthritis, complex regional pain syndrome (RSD), relapsing polychondritis, restless legs syndrome (RLS), rheumatic fever, rheumatoid arthritis, sarcoidosis, autoimmune polyendocrine syndrome type 2, scleritis, scleroderma, serpiginous choroiditis, Sjogren's syndrome, stiff-person syndrome (SPS), small fiber neuropathy The term "Clinical Conditions" refers to diseases or conditions including, but not limited to, Qi, systemic lupus erythematosus (SLE), Susac syndrome, Sydenham's chorea, sympathetic nerve fibers, Takayasu's arteritis, orchitis, Tolosa-Hunt syndrome, transverse myelitis (TM), tubulointerstitial nephritis-uveitis syndrome (TINU), ulcerative colitis (UC), undifferentiated connective tissue disease, uveitis, vasculitis, leukoplakia, and Vogt-Koyanagi-Harada disease, and also applies to arteriosclerosis caused by said diseases. The inflammatory response-related mechanism of p38 is not only involved in the aforementioned diseases caused by innate or acquired immunity, but also in acute inflammation caused by infectious agents, i.e., Ebola virus disease, Marburg fever, Lassa fever, Crimean-Congo hemorrhagic fever, South American hemorrhagic fever, Rift Valley fever, new infectious diseases including COVID-19, severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), human infections caused by animal-derived influenza, new influenza, chickenpox, measles, typhoid fever, paratyphoid fever, enterohemorrhagic Escherichia coli infection, Hepatitis A infection, whooping cough, mumps, etc.Rubella, meningococcal infection, Haemophilus influenzae type b infection, pneumococcal infection, scarlet fever, vancomycin-resistant Staphylococcus aureus (VRSA) infection, carbapenem-resistant Enterobacteriaceae (CRE) infection, type E infection, tetanus, Japanese encephalitis, Legionnaires' disease, Vibrio septicemia, typhus, murine typhus, scrub typhus, leptospirosis, brucellosis, rabies, hemorrhagic fever with renal syndrome, yellow fever, dengue fever, Q fever, West Nile fever, Lyme disease, tick-borne encephalitis, and chikungunya The present invention is comprehensively applicable to infectious, congenital, or acquired inflammatory diseases occurring in subjects, including, but not limited to, flu-like fever, severe fever with thrombocytopenia syndrome (SFTS), vancomycin-resistant enterococcus (VRE) infection, methicillin-resistant Staphylococcus aureus (MRSA) infection, multidrug-resistant Pseudomonas aeruginosa (MRPA) infection, multidrug-resistant Acinetobacter (MRAB) infection, intestinal infection, acute respiratory infection, enterovirus infection, and human papillomavirus infection.

[0121] The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier or excipient.

[0122] The active ingredient of the present invention can be administered alone or in admixture with a suitable carrier, etc., and such dosage form may be a single dose or multiple doses. The pharmaceutical composition may be a solid or liquid formulation. Solid formulations include, but are not limited to, powders, granules, tablets, capsules, suppositories, etc. Solid formulations may contain, but are not limited to, carriers, flavoring agents, binders, preservatives, disintegrants, lubricants, fillers, etc. Liquid formulations include, but are not limited to, solutions such as water and propylene glycol solutions, suspensions, emulsifiers, etc., and may be prepared by adding appropriate colorants, flavoring agents, stabilizers, thickeners, etc. For example, powders can be prepared by simply mixing the active ingredient of the present invention with a suitable pharmaceutically acceptable carrier, such as lactose, starch, or amorphous cellulose. Granules can be prepared by mixing the active ingredient of the present invention, a suitable pharmaceutically acceptable carrier, and a suitable pharmaceutically acceptable binder such as polyvinylpyrrolidone, hydroxypropyl cellulose, etc., followed by wet granulation using a solvent such as water, ethanol, isopropanol, etc., or by dry granulation using compression force. Tablets can be prepared by mixing the granules with a suitable pharmaceutically acceptable lubricant such as magnesium stearate, followed by tableting using a tablet press.

[0123] The pharmaceutical composition of the present invention can be administered orally, by injection (e.g., intramuscular injection, intraperitoneal injection, intravenous injection, infusion, subcutaneous injection, implant), inhalant, nasal administration, vaginal administration, rectal administration, sublingual administration, transdermal administration, topical administration, etc., depending on the disease to be treated and the condition of the individual, but is not limited thereto. Depending on the administration route, it can be formulated into an appropriate dosage unit form containing commonly used, non-toxic pharmaceutically acceptable carriers, additives, and vehicles.

[0124] The pharmaceutical composition of the present invention can be administered daily at a dose of about 0.0001 mg / kg to about 10 g / kg, or at a daily dosage of about 0.001 mg / kg to about 1 g / kg. However, the dosage may vary depending on the degree of purification of the mixture, the patient's condition (age, sex, weight, etc.), the severity of the condition being treated, etc. If necessary, for convenience, the total daily dosage can be administered in several divided doses throughout the day.

[0125] The present invention will be described in detail below with reference to examples.

[0126] However, the following examples are provided to specifically illustrate the present invention, and the scope of the present invention is not limited to the following examples.

[0127] [Manufacturing example] First, the compounds represented by Formulas I to IV of the present invention were designed and synthesized as follows. Some of the materials used in the synthesis process are abbreviated as follows: ACN: acetonitrile, DCM: dichloromethane, HCl: hydrochloride, EtOAc: ethyl acetate, MeOH: methanol, EtOH: ethanol, DCE: dichloroethane, DMSO: dimethyl sulfoxide, DMF: N,N-dimethylformamide, THF: tetrahydrofuran, TBSCl: tert-butyldimethylsilyl chloride, TBAI: tetrabutylammonium iodide, DEA: diethylamine, TEA: triethylamine, DIPEA: N,N-diisopropylethylamine, NIS: n-iodosuccinimide, PTSA: p-toluenesulfonic acid, TFA: trifluoroacetic acid acid, BINAP:(2,2'-bis(diphenylphosphino)-1,1'-binaphthyl), NBS:n-bromosuccinimide, EDC:1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, HATU:1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate, HOBt:1-hydroxybenzotriazole hydrate, DCC:N,N'-dicyclohexylcarbodiimide, DMAP:4-dimethylaminopyridine and mCPBA:meta-chloroperoxybenzoic acid.

[0128] [Manufacturing example I] Compounds according to Formula I of the present invention were synthesized as follows.

[0129] [Production Example 1-1] 4-(3-aminoprop-1-yn-1-yl)aniline hydrochloride

[0130] TIFF2025503429000206.tif26170

[0131] Step 1: tert-butyl(3-(4-aminophenyl)prop-2-yn-1-yl)carbamate (Intermediate 1-1a)

[0132] TIFF2025503429000207.tif28170

[0133] 4-iodoaniline (1.0 mmol, 1.0 eq), n-boc propargylamine (1.5 eq), PD(PPh3)4 (0.01 eq), CuI (0.03 eq), DEA (1.5 eq), and 3.0 mL of Et2O were mixed, and the gas in the reaction vessel was replaced with Ar gas. The mixture was then stirred overnight at room temperature. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column chromatography to synthesize intermediate 1-1a.

[0134] Step 2: 4-(3-aminoprop-1-yn-1-yl)aniline hydrochloride

[0135] Intermediate 1-1a (0.5 mmol, 1.0 eq) synthesized in Step 1 was dissolved in 3.0 mL of DCM, and then HCl solution (4.0 M in dioxane, 2.0 eq) was added dropwise. After 1 hour, the resulting solid was filtered and dried to synthesize Example 1-1 without further purification.

[0136] 1H NMR (500 MHz, DMSO-d6) δ 8.41 (s, 3H), 7.24 (d, J = 8.4 Hz, 2H), 6.83 (d, J = 8.1 Hz, 2H), 3.89 (q, J = 5.6 Hz, 2H)

[0137] [Production Example 1-3] 3-(3-aminoprop-1-yn-1-yl)aniline hydrochloride

[0138] TIFF2025503429000208.tif23170

[0139] Intermediate 1-3a (tert-butyl(3-(3-aminophenyl)prop-2-yn-1-yl)carbamate) was synthesized from 3-iodoaniline in the same manner as in Production Example 1-1, and then Example 1-3 was synthesized.

[0140] 1 H NMR (500 MHz, DMSO-d6) δ 8.57 (s, 3H), 7.42 (d, J = 7.4 Hz, 1H), 7.29 (dd, J = 24.4, 14.7 Hz, 3H), 3.99 (t, J = 5.5 Hz, 2H)

[0141] [Production Example 3-1] methyl 5-amino-2-(3-aminoprop-1-yn-1-yl)benzoate hydrochloride

[0142] TIFF2025503429000209.tif28170

[0143] Intermediate 3-1a (methyl 5-amino-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate) was synthesized from methyl 5-amino-2-iodobenzoate in the same manner as in Production Example 1-1, and then Example 3-1 was synthesized.

[0144] 1 H NMR (500 MHz, DMSO-d6) δ 8.42 (s, 3H), 7.26 (d, J = 8.4 Hz, 1H), 7.13 (d, J = 2.4 Hz, 1H), 6.82 (dd, J = 8.4, 2.4 Hz, 1H), 3.90 (q, J = 5.7 Hz, 2H), 3.79 (s, 3H)

[0145] [Production Example 3-25] methyl 4-amino-2-(3-aminoprop-1-yn-1-yl)benzoate hydrochloride

[0146] TIFF2025503429000210.tif25170

[0147] Intermediate 3-25a (methyl 4-amino-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate) was synthesized from methyl 4-amino-2-iodobenzoate in the same manner as in Production Example 1-1, and then Example 3-25 was synthesized.

[0148] 1 H NMR (500 MHz, DMSO-d6) δ 8.47 (s, 3H), 7.67 (d, J = 8.7 Hz, 1H), 6.72 (dd, J = 4.9, 2.3 Hz, 1H), 6.62 (ddd, J = 8.7, 4.3, 2.2 Hz, 1H), 3.99 (dd, J = 5.8, 3.0 Hz, 2H), 3.74 (s, 3H)

[0149] [Production Example 3-49] 2-amino-4-(4-aminophenyl)but-3-yn-1-ol hydrochloride

[0150] TIFF2025503429000211.tif33170

[0151] Step 1: tert-butyl 4-ethynyl-2,2-dimethyloxazolidine-3-carboxylate (Intermediate 3-49a)

[0152] TIFF2025503429000212.tif22170

[0153] tert-Butyl 4-formyl-2,2-dimethyloxazolidine-3-carboxylate (8.7 mmol, 1.0 eq) and dimethyl-1-diazo-2-oxopropyl phosphonate (1.5 eq) were dissolved in MeOH, and then K2CO3 (2.0 eq) was added at 0°C. The mixture was stirred at 0°C for 1 hour and then at room temperature overnight. The reaction mixture was extracted with n-hexane and HO, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column to synthesize the compound.

[0154] Step 2: tert-butyl 4-((4-aminophenyl)ethynyl)-2,2-dimethyloxazolidine-3-carboxylate (Intermediate 3-49b)

[0155] TIFF2025503429000213.tif40170

[0156] Intermediate 3-49a (4.4 mmol, 1.0 eq) synthesized in Step 1, 4-iodoaniline (1.0 eq), PdCl2(PPh3)2 (0.1 eq), and CuI (0.1 eq) were dissolved in 40.0 mL of DEA and stirred overnight at room temperature. The reaction mixture was distilled under reduced pressure and purified via flash column chromatography to obtain the desired compound.

[0157] Step 3: 2-amino-4-(4-aminophenyl)but-3-yn-1-ol hydrochloride

[0158] Example 3-49 was synthesized from intermediate 3-49b synthesized in Step 3 in the same manner as in Step 2 of Production Example 1-1.

[0159] 1 H NMR (500 MHz, DMSO-d6) δ 7.98 (s, 3H), 7.38 (d, J = 8.4 Hz, 2H), 6.63 - 6.61 (m, 2H), 3.60 - 3.57 (m, 2H), 3.51 (d, J = 4.9 Hz, 1H)

[0160] [Production Example 5-116] methyl 2,4-bis(3-aminoprop-1-yn-1-yl)benzoate dihydrochloride

[0161] TIFF2025503429000214.tif23170

[0162] Step 1: methyl 4-bromo-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate (Intermediate 5-116a)

[0163] TIFF2025503429000215.tif23170

[0164] Intermediate 5-116a was synthesized from methyl 4-bromo-2-iodobenzoate in the same manner as in Step 1 of Production Example 1-1.

[0165] Step 2: methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoate (intermediate 5-116b)

[0166] TIFF2025503429000216.tif29170

[0167] Intermediate 5-116a (0.3 mmol, 1.0 eq), n-propargylphthalimide (1.5 eq), Pd(PPh3)4 (0.1 eq), CuI (0.3 eq), TBAI (3.0 eq), 3.0 mL of DMF, and 1.0 mL of TEA were mixed and the gas in the reaction vessel was replaced with Ar gas. The mixture was then heated to 110 °C for 20 min under MW. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column to give the desired compound.

[0168] Step 3: methyl 4-(3-aminoprop-1-yn-1-yl)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate (Intermediate 5-116c)

[0169] TIFF2025503429000217.tif23170

[0170] Intermediate 5-116b (0.17 mmol, 1.0 eq), hydrazine monohydrate (3.0 eq), MeOH 0.8 mL, and HO 0.2 mL were mixed and stirred at 50° C. for 2 hours. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over NaSO and then evaporated under reduced pressure to give the compound without further purification.

[0171] Step 4: methyl 2,4-bis(3-aminoprop-1-yn-1-yl)benzoate dihydrochloride

[0172] TIFF2025503429000218.tif23170

[0173] Example 5-116 was synthesized from Intermediate 5-116c in the same manner as in Step 2 of Preparation Example 1-1.

[0174] 1H NMR (500 MHz, DMSO-d6) δ 8.61 (d, J = 14.6 Hz, 6H), 7.93 - 7.88 (m, 1H), 7.61 (d, J = 1.7 Hz, 1H), 7.57 (dd, J = 8.2, 1.7 Hz, 1H), 3.98 (dd, J = 5.9, 3.0 Hz, 4H), 3.84 (s, 3H).

[0175] [Manufacturing example II] The compound of the present invention corresponding to formula II was synthesized as follows.

[0176] [Production Example 4-162] 4-amino-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)butanamide dihydrochloride

[0177] TIFF2025503429000219.tif17170

[0178] Example 4-162 was synthesized from 4-((tert-Butoxycarbonyl)amino)butanoic acid and intermediate 1-1a in the same manner as in Production Example 1-1'.

[0179] 1 H NMR (500 MHz, DMSO-d6) δ 10.46 (s, 1H), 8.45 (s, 3H), 7.99 (s, 3H), 7.68 - 7.62 (m, 2H), 7.38 - 7.32 (m, 2H), 3.92 (q, J = 5.6 Hz, 2H), 2.78 (h, J = 6.2 Hz, 2H), 2.43 (d, J = 7.2 Hz, 2H), 1.83 (p, J = 7.3 Hz, 2H).

[0180] [Production Example 4-166] 5-(3-aminoprop-1-yn-1-yl)-N-(3-aminopropyl)furan-2-carboxamide dihydrochloride

[0181] TIFF2025503429000220.tif17170

[0182] Example 4-166 was synthesized from tert-Butyl(3-aminopropyl)carbamate and intermediate 4-227b in the same manner as in Preparation Example 1-1′.

[0183] 1 H NMR (500 MHz, DMSO-d6) δ 8.79 (t, J = 5.9 Hz, 1H), 8.67 (s, 3H), 8.00 (s, 3H), 7.21 (d, J = 3.6 Hz, 1H), 6.96 (d, J = 3.6 Hz, 1H), 4.02 (q, J = 5.6 Hz, 2H), 3.24 (q, J = 6.5 Hz, 2H), 2.75 (dq, J = 11.9, 6.2 Hz, 2H), 1.78 - 1.71 (m, 2H).

[0184] Furthermore, for use in a biotin-based pulldown assay, biotin-conjugated compound 4-166 was prepared as follows.

[0185] 5-(3-aminoprop-1-yn-1-yl)-N-(3-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)propyl)furan-2-carboxamide hydrochloride

[0186] TIFF2025503429000221.tif43170

[0187] Step 1: tert-butyl(3-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)propyl)carbamate

[0188] TIFF2025503429000222.tif38170

[0189] The intermediate tert-butyl(3-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)propyl)carbamate was synthesized from biotin and tert-butyl(3-aminopropyl)carbamate in the same manner as in Step 1 of Preparation Example 1-1'.

[0190] Step 2: tert-butyl(3-(5-((3-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)propyl)carbamoyl)furan-2-yl)prop-2-yn-1-yl)carbamate

[0191] TIFF2025503429000223.tif44170

[0192] The intermediate tert-butyl(3-(5-((3-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)propyl)carbamoyl)furan-2-yl)prop-2-yn-1-yl)carbamate was synthesized in the same manner as in Step 1 of Production Example 1-1′.

[0193] Step 3: 5-(3-aminoprop-1-yn-1-yl)-N-(3-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)propyl)furan-2-carboxamide hydrochloride

[0194] A biotin-bound 4-166 compound was synthesized from the intermediate of Step 2 in the same manner as in Step 2 of Production Example 1-1.

[0195] 1H NMR (500 MHz, DMSO-d6) δ 8.61 - 8.48 (m, 4H), 7.86 (t, J = 5.7 Hz, 1H), 7.19 (d, J = 3.6 Hz, 1H), 7.00 (d, J = 3.6 Hz, 1H), 6.44 (s, 1H), 4.30 (dd, J = 7.7, 4.6 Hz, 1H), 4.14 - 4.10 (m, 1H), 4.08 (q, J = 5.6 Hz, 2H), 3.20 (q, J = 6.2 Hz, 2H), 3.12 - 3.03 (m, 3H), 2.81 (dd, J = 12.4, 5.1Hz, 1H), 2.57 (d, J = 12.4 Hz, 1H), 2.06 (t, J = 7.4 Hz, 2H), 1.63 - 1.56 (m, 3H), 1.54 - 1.43 (m, 3H), 1.33 - 1.25 (m, 2H), MS m / z [M+H] + 448.43.

[0196] [Production Example 4-228] methyl 4-(4-aminobutanamido)-2-(3-aminoprop-1-yn-1-yl)benzoate dihydrochloride

[0197] TIFF2025503429000224.tif21170

[0198] Example 4-228 was synthesized from 4-((tert-Butoxycarbonyl)amino)butanoic acid and intermediate 3-25a in the same manner as in Preparation Example 1-1' via intermediate 4-228a (methyl 4-(4-aminobutanamido)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate).

[0199] 1H NMR (500 MHz, DMSO-d6) δ 10.92 (s, 1H), 8.60 (s, 3H), 8.13 (s, 3H), 8.08 (d, J = 2.2 Hz, 1H), 7.88 (d, J = 8.7 Hz, 1H), 7.71 (dd, J = 8.7, 2.2 Hz, 1H), 4.07 - 3.99 (m, 2H), 3.83 (s, 3H), 2.96 (dt, J = 9.2, 4.6 Hz, 2H), 2.82 (q, J = 6.7 Hz, 2H), 2.53 (t, J = 7.3 Hz, 2H).

[0200] Furthermore, for use in a biotin-based pulldown assay, a biotin-conjugated 4-228 compound was prepared as follows.

[0201] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)butanamido)benzoate hydrochloride

[0202] TIFF2025503429000225.tif33170

[0203] Step 1: methyl 4-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)butanamido)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate

[0204] TIFF2025503429000226.tif31170

[0205] Intermediate 3-25a (0.2 mmol, 1.0 eq), 4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)butanoic acid (1.0 eq), and HATU (1.1 eq) were dissolved in dry DMF (0.7 mL), and the gas in the reaction vessel was replaced with Ar gas. TEA (2.0 eq) was added dropwise, and the mixture was stirred at room temperature for 6 hours. The reaction mixture was purified via flash column to synthesize the intermediate methyl 4-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)butanamido)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate.

[0206] Step 2: methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(4-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)butanamido)benzoate

[0207] TIFF2025503429000227.tif35170

[0208] The intermediate (0.06 mmol, 1.0 eq) prepared in Step 1 was dissolved in 0.3 mL of piperidine / DMF (1:4) and stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, and biotin (1.0 eq) was added. The intermediate methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(4-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)butanamido)benzoate was synthesized in the same manner as in Step 1 above.

[0209] Step 3: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)butanamido)benzoate hydrochloride

[0210] From the intermediate produced in Step 2, a biotin-bound 4-228 compound was synthesized in the same manner as in Step 2 of Production Example 1-1.

[0211] 1 H NMR (500 MHz, DMSO-d6) δ 10.43 (s, 1H), 8.36 (s, 3H), 8.13 (d, J = 2.2 Hz, 1H), 7.89 (d, J = 8.7 Hz, 1H), 7.85 (t, J = 5.5 Hz, 1H), 7.55 (dd, J = 8.7, 2.2 Hz, 1H), 6.43 (s, 1H), 6.38 (s, 1H), 4.29 (dd, J = 7.6, 4.7 Hz, 1H), 4.14 - 4.08 (m, 1H), 4.06 (q, J = 5.6 Hz, 2H), 3.83 (s, 3H), 3.10 - 3.05 (m, 3H), 2.80 (dd, J = 12.4, 5.1 Hz, 1H), 2.56 (d, J = 12.6 Hz, 1H), 2.38 - 2.32 (m, 3H), 2.05 (t, J = 7.4 Hz, 2H), 1.73 - 1.67 (m, 2H), 1.62 - 1.56 (m, 1H), 1.53 - 1.46 (m, 2H), 1.31 - 1.26 (m, 2H), MS m / z [M+H] + 516.47.

[0212] [Production Example 5-8] 4-amino-N-(4-(3-aminoprop-1-yn-1-yl)-3-(hydroxymethyl)phenyl)butanamide trifluoroacetic acid

[0213] TIFF2025503429000228.tif21170

[0214] 4-((tert-Butoxycarbonyl)amino)butanoic acid and intermediate 3-27c were synthesized in the same manner as in Step 5 of Preparation Example 3-3' and Step 2 of Preparation Example 1-1, and then further purified by prep HPLC to synthesize Example 5-8.

[0215] 1 H NMR (500 MHz, DMSO-d6) δ 7.62 (d, J = 2.2 Hz, 1H), 7.44 (dd, J = 8.4, 2.2 Hz, 1H), 7.35 (d, J = 8.3 Hz, 1H), 4.58 (s, 2H), 3.91 (s, 2H), 2.84 (t, J = 7.7 Hz, 2H), 2.40 (d, J = 7.3 Hz, 2H), 1.86 - 1.82 (m, 2H); MS m / z [M+Na] + 284.30.

[0216] [Production Example 5-18] methyl 2-(3-aminoprop-1-yn-1-yl)-4-((3-aminopropyl)carbamoyl)benzoate dihydrochloride

[0217] TIFF2025503429000229.tif30170

[0218] Step 1: methyl 2-bromo-4-((3-((tert-butoxycarbonyl)amino)propyl)carbamoyl)benzoate(5-18a)

[0219] TIFF2025503429000230.tif18170

[0220] Intermediate 5-18a was synthesized from tert-Butyl(3-aminopropyl)carbamate and 3-bromo-4-(methoxycarbonyl)benzoic acid in the same manner as in Step 5 of Preparation Example 4-1′.

[0221] Step 2: methyl 2-(3-aminoprop-1-yn-1-yl)-4-((3-aminopropyl)carbamoyl)benzoate dihydrochloride

[0222] Example 5-18 was synthesized from Intermediate 5-18a in the same manner as in Production Example 1-1.

[0223] 1 H NMR (500 MHz, DMSO-d6) δ 9.01 (s, 1H), 8.52 (s, 3H), 8.09 - 8.08 (m, 1H), 8.01 - 7.99 (m, 2H), 7.95 (s, 3H), 4.08 - 4.03 (m, 2H), 3.90 (s, 3H), 3.35 - 3.29 (m, 2H), 2.86 - 2.80 (m, 2H), 1.82 (p, J = 6.9 Hz, 2H).

[0224] [Production Example 5-81] 4-amino-N-(3-(3-aminoprop-1-yn-1-yl)-4-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)butanamide dihydrochloride

[0225] TIFF2025503429000231.tif21170

[0226] Example 5-81 was synthesized from 4-((tert-Butoxycarbonyl)amino)butanoic acid and intermediate 5-81c in the same manner as in Production Example 1-1'.

[0227] 1H NMR (500 MHz, DMSO-d6) δ 10.69 (s, 1H), 9.30 (s, 2H), 8.38 (s, 3H), 8.16 (d, J = 2.2 Hz, 1H), 7.97 (s, 3H), 7.77 (dd, J = 8.6, 2.2 Hz, 1H), 7.66 (d, J = 8.5 Hz, 1H), 3.94 (s, 2H), 2.85 (s, 2H), 2.53 (s, 2H), 1.90 - 1.87 (m, 2H).

[0228] [Production Example 5-109] 11-((3-(3-aminoprop-1-yn-1-yl)phenyl)amino)-11-oxoundecanoic acid hydrochloride

[0229] TIFF2025503429000232.tif16170

[0230] Undecanedioic acid, Intermediate 1-3a, Preparation Example 5-77, Preparation Example 2, and Synthesis Example 5-109.

[0231] 1 H NMR (500 MHz, DMSO-d6) δ 10.15 (s, 1H), 8.45 (s, 3H), 7.96 - 7.91 (m, 1H), 7.53 - 7.49 (m, 1H), 7.33 (t, J = 7.9 Hz, 1H), 7.11 (dt, J = 7.7, 1.2 Hz, 1H), 3.99 (s, 2H), 2.34 - 2.24 (m, 4H), 1.61 - 1.44 (m, 4H), 1.30 - 1.19 (m, 10H).

[0232] [Production Example 5-110] 11-((3-(3-aminoprop-1-yn-1-yl)-4-(methoxycarbonyl)phenyl)amino)-11-oxoundecanoic acid hydrochloride

[0233] TIFF2025503429000233.tif21170

[0234] Example 5-110 was synthesized from undecanedioic acid and intermediate 3-25a in the same manner as in Step 2 of Preparation Example 5-77'.

[0235] 1 H NMR (500 MHz, DMSO-d6) δ 10.43 (s, 1H), 8.85 (s, 1H), 8.41 (s, 3H), 8.13 (d, J = 2.2 Hz, 1H), 7.89 (d, J = 8.7 Hz, 1H), 7.56 (dd, J = 8.7, 2.2 Hz, 1H), 4.04 (s, 2H), 3.83 (s, 3H), 2.34 (t, J = 7.4 Hz, 2H), 2.25 (d, J = 7.4 Hz, 2H), 1.57 (t, J = 7.0 Hz, 2H), 1.54 - 1.42 (m, 12H).

[0236] [Production Example 5-150] methyl 4-(4-amino-N-methylbutanamido)-2-(3-aminoprop-1-yn-1-yl)benzoate hydrochloride

[0237] TIFF2025503429000234.tif32170

[0238] Example 5-150 was synthesized from 4-Aminobutanoic acid and intermediate 5-150a in the same manner as in Step 2 of Preparation Example 5-77'.

[0239] 1H NMR (500 MHz, DMSO-d6) δ 8.55 (s, 3H), 7.95 - 7.93 (m, 1H), 7.90 (s, 3H), 7.63 - 7.59 (m, 1H), 7.54 - 7.51 (m, 1H), 4.06 - 4.00 (m, 2H), 3.88 (s, 3H), 3.23 (s, 3H), 2.95 - 2.84 (m, 2H), 2.81 - 2.72 (m, 2H), 1.81 - 1.73 (m, 2H).

[0240] [Manufacturing example III] The compound of the present invention corresponding to formula III was synthesized as follows.

[0241] [Production Example 4-2] methyl 2-(3-aminoprop-1-yn-1-yl)-5-(piperazin-1-yl)benzoate hydrochloride

[0242] TIFF2025503429000235.tif15170

[0243] Example 4-2 was synthesized from methyl 2-bromo-5-fluorobenzoate in the same manner as in Production Example 5-7.

[0244] 1 H NMR (500 MHz, DMSO-d6) δ 9.40 (s, 2H), 8.46 (s, 3H), 7.42 (d, J = 8.7 Hz, 1H), 7.34 (d, J = 2.7 Hz, 1H), 7.18 (dd, J = 8.8, 2.8 Hz, 1H), 3.93 (q, J = 5.7 Hz, 2H), 3.82 (s, 3H), 3.47 (s, 4H), 3.15 (s, 4H)

[0245] [Production Example 5-7] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(piperazin-1-yl)benzoate hydrochloride

[0246] TIFF2025503429000236.tif21170

[0247] Step 1: methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-fluorobenzoate (Intermediate 5-7a)

[0248] TIFF2025503429000237.tif22170

[0249] Intermediate 5-7a was synthesized from methyl 2-bromo-4-fluorobenzoate in the same manner as in Step 2 of Preparation Example 4-179′.

[0250] Step 2: methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(piperazin-1-yl)benzoate (Intermediate 5-7b)

[0251] TIFF2025503429000238.tif21170

[0252] Intermediate 5-7a (0.6 mmol, 1.0 eq), piperazine (3.0 eq), K2CO3 (1.5 eq), and 2.0 mL of DMSO were mixed, and the gas in the reaction vessel was replaced with Ar gas. After blocking external light, the mixture was stirred at 95 °C for 18 hours. The reaction mixture was extracted with EtOAc and H2O, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column to synthesize intermediate 5-7b.

[0253] Step 3: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(piperazin-1-yl)benzoate hydrochloride

[0254] Example 5-7 was synthesized from Intermediate 5-7b in the same manner as in Step 2 of Preparation Example 1-1.

[0255] 1 H NMR (500 MHz, DMSO-d6) δ 9.35 (s, 2H), 8.50 (s, 3H), 7.79 (d, J = 8.9 Hz, 1H), 7.10 (d, J = 2.7 Hz, 1H), 7.06 (dd, J = 8.9, 2.7 Hz, 1H), 3.97 (q, J = 5.6 Hz, 2H), 3.77 (s, 3H), 3.53 (t, J = 5.3 Hz, 4H), 3.14 (d, J = 13.1 Hz, 4H).

[0256] [Production Example 5-23] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(piperidine-4-carboxamido)benzoate hydrochloride

[0257] TIFF2025503429000239.tif31170

[0258] Example 5-23 was synthesized from 1-(tert-Butoxycarbonyl)piperidine-4-carboxylic acid and intermediate 3-25a in the same manner as in Preparation Example 1-1′.

[0259] 1H NMR (500 MHz, DMSO-d6) δ 10.85 (s, 1H), 9.04 (d, J = 11.2 Hz, 1H), 8.78 (d, J = 11.0 Hz, 1H), 8.47 (s, 3H), 8.06 (d, J = 2.2 Hz, 1H), 7.86 (d, J = 8.7 Hz, 1H), 7.64 (dd, J = 8.7, 2.2 Hz, 1H), 4.00 (q, J = 5.6 Hz, 2H), 3.80 (s, 3H), 3.31 - 3.23 (m, 2H), 2.86 (q, J = 11.7 Hz, 2H), 2.72 (tt, J = 11.1, 3.8 Hz, 1H), 1.94 (d, J = 13.4 Hz, 2H), 1.83 - 1.74 (m, 2H).

[0260] [Production Example 5-28] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(3-(piperidine-4-carboxamido)prop-1-yn-1-yl)benzoate dihydrochloride

[0261] TIFF2025503429000240.tif26170

[0262] Example 5-28 was synthesized from 1-(tert-Butoxycarbonyl)piperidine-4-carboxylic acid and intermediate 5-116c in the same manner as in Preparation Example 1-1′.

[0263] 11H NMR (500 MHz, DMSO-d6) δ 8.56 (dd, J = 12.3, 6.8 Hz, 4H), 7.88 - 7.84 (m, 1H), 7.57 (d, J = 1.8 Hz, 1H), 7.51 (dd, J = 8.2, 1.7 Hz, 1H), 4.11 (d, J = 5.4 Hz, 2H), 3.98 (q, J = 5.6 Hz, 2H), 3.83 (s, 3H), 3.21 (d, J = 12.5 Hz, 2H), 2.89 - 2.77 (m, 2H), 2.42 (dt, J = 11.0, 3.9 Hz, 1H), 1.85 - 1.80 (m, 2H), 1.75 - 1.68 (m, 2H).

[0264] [Production Example 5-29] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)benzoate dihydrochloride

[0265] TIFF2025503429000241.tif34170

[0266] Step 1: (9H-fluoren-9-yl)methyl 4-(bromomethyl)piperidine-1-carboxylate (Intermediate 5-29a)

[0267] TIFF2025503429000242.tif11170

[0268] 4-(Bromomethyl)piperidine hydrochloride (1.0 eq), (9H-fluoren-9-yl)methyl carbonochloridate (1.2 eq), pyridine (1.5 eq), and dry DCM were mixed and stirred at room temperature for 2 hours. The reaction mixture was extracted with EtOAc and H2O, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified by flash column chromatography to synthesize intermediate 5-29a.

[0269] Step 2: (9H-fluoren-9-yl)methyl 4-(piperazin-1-ylmethyl)piperidine-1-carboxylate (Intermediate 5-29b)

[0270] TIFF2025503429000243.tif21170

[0271] Intermediate 5-29a (1.0 eq), piperazine (9.0 eq), and dry ACN were mixed and stirred at 80 °C overnight. The reaction mixture was extracted with EtOAc and H2O into an organic layer, which was then dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified using a flash column to synthesize intermediate 5-29b.

[0272] Step 3: (9H-fluoren-9-yl)methyl 4-((4-(3-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(methoxycarbonyl)phenyl)piperazin-1-yl)methyl)piperidine-1-carboxylate (Intermediate 5-29c)

[0273] TIFF2025503429000244.tif32170

[0274] Intermediate 5-29b (1.0 eq), Intermediate 5-116a (1.0 eq), RuPhos Pd G3 (0.1 eq), CS2CO3 (3.0 eq), and 1.5 mL of dioxane were mixed and stirred at 100 °C overnight. The reaction mixture was extracted with EtOAc and H2O into an organic layer, which was then dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified by flash column chromatography to give Intermediate 5-29c.

[0275] Step 4: methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)benzoate (intermediate 5-29d)

[0276] TIFF2025503429000245.tif34170

[0277] Intermediate 5-29c (1.0 eq), DEA (1.0 mL), and THF (1.0 mL) were mixed and stirred at room temperature for 1 hour. The reaction mixture was distilled under reduced pressure and purified through a flash column to obtain intermediate 5-29d.

[0278] Step 5: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)benzoate hydrochloride

[0279] Example 5-29 was synthesized from Intermediate 5-29d in the same manner as in Step 2 of Preparation Example 1-1.

[0280] 1H NMR (500 MHz, DMSO-d6) δ 9.12 - 8.84 (m, 2H), 8.60 (s, 3H), 7.83 (d, J = 8.9 Hz, 1H), 7.20 (d, J = 2.7 Hz, 1H), 7.13 (dd, J = 9.0, 2.7 Hz, 1H), 3.99 (s, 2H), 3.81 (s, 3H), 3.54 - 3.45 (m, 6H), 3.26 (d, J = 12.6 Hz, 2H), 3.08 (s, 4H), 2.85 (q, J = 11.8 Hz, 2H), 2.18 (s, 1H), 2.07 - 2.00 (m, 2H), 1.45 (d, J = 12.3 Hz, 2H).

[0281] [Production Example 5-39] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(2,7-diazaspiro[3.5]nonane-7-carbonyl)benzoate dihydrochloride

[0282] TIFF2025503429000246.tif21170

[0283] Example 5-39 was synthesized in the same manner as in Preparation Example 5-41 using tert-Butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate.

[0284] 1 H NMR (500 MHz, DMSO-d6) δ 8.49 (s, 3H), 7.92 (d, J = 8.0 Hz, 1H), 7.86 (d, J = 8.0 Hz, 1H), 7.28 (d, J = 1.7 Hz, 1H), 4.00 (s, 2H), 3.85 (s, 3H), 3.68 (s, 8H), 3.53 (s, 4H).

[0285] [Production Example 5-40] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(2,6-diazaspiro[3.3]heptan-2-yl)benzoate dihydrochloride

[0286] TIFF2025503429000247.tif21170

[0287] Step 1: tert-butyl 6-(3-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(methoxycarbonyl)phenyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (Intermediate 5-40a)

[0288] TIFF2025503429000248.tif29170

[0289] Intermediate 5-35a (0.5 mmol, 1.0 eq), tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (1.0 eq), SPhos Pd G3 (0.1 eq), CS2CO3 (3.0 eq), and 1.5 mL of dioxane were mixed and stirred at 100 °C overnight. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column chromatography to give Intermediate 5-40a.

[0290] Step 2: methyl 2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(2,6-diazaspiro[3.3]heptan-2-yl)benzoate trifluoroacetic acid (intermediate 5-40b)

[0291] TIFF2025503429000249.tif28170

[0292] Example 5-40b was synthesized from Intermediate 5-40a in the same manner as in Step 4 of Preparation Example 4-1′.

[0293] Step 3: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(2,6-diazaspiro[3.3]heptan-2-yl)benzoate dihydrochloride

[0294] Example 5-40 was synthesized from Intermediate 5-40b by the same methods as Step 3 of Preparation Example 5-116 and Step 2 of Preparation Example 1-1 in this order.

[0295] 1 H NMR (500 MHz, DMSO- d6) δ 9.65 (s, 2H), 8.71 (s, 3H), 7.31 (d, J = 8.7 Hz, 1H), 6.72 (d, J = 2.3 Hz, 1H), 6.67 (d, J = 9.6 Hz, 1H), 3.98 (s, 2H), 3.96 (s, 3H), 3.65 - 3.62 (m, 4H), 3.56 (s, 4H).

[0296] [Production Example 5-41] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(2,6-diazaspiro[3.3]heptane-2-carbonyl)benzoate dihydrochloride

[0297] TIFF2025503429000250.tif23170

[0298] Step 1: tert-butyl 6-(3-iodo-4-(methoxycarbonyl)benzoyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (Intermediate 5-41a)

[0299] TIFF2025503429000251.tif21170

[0300] Intermediate 5-41a was synthesized from 3-iodo-4-(methoxycarbonyl)benzoic acid and tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate in the same manner as in Step 1 of Preparation Example 4-1′.

[0301] Step 2: tert-butyl 6-(3-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(methoxycarbonyl)benzoyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (Intermediate 5-41b)

[0302] TIFF2025503429000252.tif35170

[0303] Intermediate 5-41b was synthesized from intermediate 5-41a in the same manner as in Step 2 of Preparation Example 5-85'.

[0304] Step 3: methyl 2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(2,6-diazaspiro[3.3]heptane-2-carbonyl)benzoate trifluoroacetic acid (Intermediate 5-41c)

[0305] TIFF2025503429000253.tif35170

[0306] Intermediate 5-41c was synthesized from intermediate 5-41b in the same manner as in Step 4 of Production Example 4-1′.

[0307] Step 4: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(2,6-diazaspiro[3.3]heptane-2-carbonyl)benzoate

[0308] Example 5-41 was synthesized from Intermediate 5-41c by the same methods as in Step 3 of Production Example 5-116 and Step 2 of Production Example 1-1 in that order.

[0309] 1 H NMR (500 MHz, DMSO- d6) δ 9.27 (s, 3H), 7.53 (s, 1H), 7.46 - 7.41 (m, 2H), 7.39 - 7.35 (m, 2H), 4.06 - 3.99 (m, 2H), 3.94 (s, 3H), 3.91 (s, 8H).

[0310] [Production Example 5-86] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(piperidin-4-ylamino)benzoate dihydrochloride

[0311] TIFF2025503429000254.tif25170

[0312] Step 1: methyl 4-amino-2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoate (Intermediate 5-86a)

[0313] TIFF2025503429000255.tif26170

[0314] Intermediate 5-86a was synthesized from methyl 4-amino-2-iodobenzoate in the same manner as in Step 2 of Preparation Example 5-116′.

[0315] Step 2: tert-butyl 4-((3-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(methoxycarbonyl)phenyl)amino)piperidine-1-carboxylate (Intermediate 5-86b)

[0316] TIFF2025503429000256.tif27170

[0317] Intermediate 5-86b was synthesized from intermediate 5-86a using tert-butyl 4-oxopiperidine-1-carboxylate in the same manner as in Step 1 of Preparation 5-85c.

[0318] Step 3: Methyl 2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(piperidin-4-ylamino)benzoate trifluoroacetic acid (Intermediate 5-86c)

[0319] TIFF2025503429000257.tif30170

[0320] Intermediate 5-86c was synthesized from intermediate 5-86b in the same manner as in Step 4 of Production Example 4-1′.

[0321] Step 4: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(piperidin-4-ylamino)benzoate hydrochloride

[0322] Example 5-86 was synthesized from Intermediate 5-86c by the same methods as in Step 3 of Preparation Example 5-116 and Step 2 of Preparation Example 1-1 in this order.

[0323] 11H NMR (500 MHz, DMSO-d6) δ 8.60 (s, 3H), 8.53 (s, 2H), 7.86 (d, J = 13.7 Hz, 1H), 6.79 (d, J = 50.9 Hz, 1H), 6.67 (dd, J = 24.5, 2.4 Hz, 1H), 4.02 (d, J = 7.1 Hz, 2H), 3.75 (s, 3H), 3.62 (dt, J = 10.9, 4.7 Hz, 2H), 3.31 - 3.26 (m, 2H), 3.21 (dt, J = 10.4, 6.5 Hz, 2H), 3.05 - 2.93 (m, 2H), 2.91 - 2.84 (m, 1H).

[0324] [Production Example 5-168] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(2,7-diazaspiro[3.5]nonan-2-yl)benzoate dihydrochloride

[0325] TIFF2025503429000258.tif22170

[0326] Step 1: tert-butyl 2-(3-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(methoxycarbonyl)phenyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate (Intermediate 5-168a)

[0327] TIFF2025503429000259.tif26170

[0328] Intermediate 5-35a (0.5 mmol, 1.0 eq), tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate (1.0 eq), RuPhos Pd G3 (0.1 eq), CS2CO3 (3.0 eq), and toluene (1.5 mL) were mixed and stirred at 80 °C overnight. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column chromatography to give intermediate 5-168a.

[0329] Step 2: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(2,7-diazaspiro[3.5]nonan-2-yl)benzoate dihydrochloride

[0330] From intermediate 5-168a, the same methods as in step 2 of manufacturing example 1-1 and step 3 of manufacturing example 5-116 were applied in order, and intermediate 5-168b (methyl 2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(2,7-diazaspiro[3.5]nonan-2-yl)benzoate) was used to synthesize Example 5-168.

[0331] 1 H NMR (500 MHz, DMSO-d6) δ 9.03 (d, J = 61.5 Hz, 3H), 8.54 (s, 3H), 7.67 (d, J = 8.9 Hz, 1H), 6.88 (d, J = 2.5 Hz, 1H), 6.76 (dd, J = 8.9, 2.5 Hz, 1H), 3.97 - 3.94 (m, 2H), 3.81 (s, 2H), 3.73 (s, 1H), 3.72 (s, 3H), 3.69 (s, 1H), 3.07 - 2.99 (m, 4H), 1.81 - 1.67 (m, 4H).

[0332] [Manufacturing example IV] The compound of the present invention corresponding to formula IV was synthesized as follows.

[0333] [Production Example 5-32] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(3-aminopropanoyl)piperazin-1-yl)benzoate hydrochloride

[0334] TIFF2025503429000260.tif34170

[0335] Example 5-32 was synthesized from β-alanine and intermediate 5-7b in the same manner as in Production Example 1-1′.

[0336] 1 H NMR (500 MHz, DMSO- d6) δ 8.80 (s, 3H), 8.59 (s, 3H), 7.93 (d, J = 7.5 Hz, 1H), 7.12 - 7.02 (m, 2H), 3.99 (t, J = 5.7 Hz, 2H), 3.79 (s, 3H), 3.62 (t, J = 5.3 Hz, 6H), 3.43 - 3.35 (m, 4H), 3.00 (q, J = 6.0 Hz, 2H).

[0337] [Manufacturing example I'] A TBM-UBM compound in which a compound (JQ1) exhibiting activity as a BRD4 (Bromodomain-containing protein 4) inhibitor is linked to a compound corresponding to Formula I of the present invention was synthesized as follows.

[0338] [Production Example 1-1'](S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0339] TIFF2025503429000261.tif34170

[0340] Step 1: tert-butyl(S)-(3-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)phenyl)prop-2-yn-1-yl)carbamate (Intermediate 1-1'a)

[0341] TIFF2025503429000262.tif34170

[0342] Commercially available JQ1 carboxylic acid ((S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetic acid) (1.0 mmol, 1.2 eq), intermediate 1-1a (1.0 eq), HOBt (1.5 eq), DIPEA (2.0 eq), EDC (1.5 eq), and 3 mL of dry DCM were mixed and stirred at room temperature for 16 h. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, evaporated, and purified via flash column chromatography to yield intermediate 1-1a.

[0343] Step 2: (S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0344] Example 1-1' was synthesized from Intermediate 1-1'a in the same manner as in Step 2 of Preparation Example 1-1.

[0345] 1 H NMR (500 MHz, Methanol-d4) δ 8.41 - 8.34 (m, 2H), 8.24 - 8.12 (m, 6H), 5.51 - 5.48 (m, 1H), 4.45 - 4.35 (m, 4H), 3.49 (s, 3H), 3.23 (d, J = 0.9 Hz, 3H), 2.48 (d, J = 0.8 Hz, 3H); MS m / z [M+H] + 531.35.

[0346] [Production Example 1-3'](S)-N-(3-(3-aminoprop-1-yn-1-yl)phenyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0347] TIFF2025503429000263.tif29170

[0348] Intermediate 1-3'a (tert-butyl(S)-(3-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)phenyl)prop-2-yn-1-yl)carbamate) was synthesized from Intermediate 1-3a in the same manner as in Production Example 1-1', and then Example 1-3' was synthesized.

[0349] 1H NMR (500 MHz, DMSO-d6) δ 10.53 (s, 1H), 8.39 (s, 3H), 7.89 (t, J = 1.9 Hz, 1H), 7.55 (ddd, J = 8.3, 2.2, 1.1 Hz, 1H), 7.47 - 7.44 (m, 2H), 7.41 - 7.36 (m, 2H), 7.34 (t, J = 7.9 Hz, 1H), 7.11 (dt, J = 7.9, 1.2 Hz, 1H), 4.57 (dd, J = 7.6, 6.7 Hz, 1H), 3.95 (td, J = 6.2, 5.7, 3.6Hz, 2H), 3.49 (dd, J = 7.3, 3.1 Hz, 2H), 2.58 (s, 3H), 2.39 (d, J = 0.9 Hz, 3H), 1.59 (d, J = 0.8 Hz, 3H); MS m / z [M+H] + 531.35.

[0350] [Production Example 3-1']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzoate hydrochloride

[0351] TIFF2025503429000264.tif40170

[0352] Intermediate 3-1'a (methyl(S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzoate) was synthesized from Intermediate 3-1a in the same manner as in Production Example 1-1', and then Example 3-1' was synthesized.

[0353] 1 1H NMR (500 MHz, DMSO-d6) δ 10.79 (s, 1H), 8.44 (s, 3H), 8.26 (d, J = 2.2 Hz, 1H), 7.86 (dd, J = 8.6, 2.3 Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.50 - 7.47(m, 2H), 7.45 - 7.41 (m, 2H), 4.64 (d, J = 7.8 Hz, 1H), 3.99 (d, J = 5.8 Hz, 2H), 3.56 (s, 3H), 2.63 (s, 3H), 2.43 (s Hz, 3H), 1.63 (s, 3H); MS m / z [M+H] + 587.35.

[0354] [Production Example 3-2'] (S)-2-(3-aminoprop-1-yn-1-yl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzoic acid hydrochloride

[0355] TIFF2025503429000265.tif40170

[0356] Step 1: (S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzoic acid (Intermediate 3-2'a)

[0357] TIFF2025503429000266.tif34170

[0358] Intermediate 3-1'a (0.1 mmol, 1.0 eq), LiOH·HO (3.0 eq), 0.1 mL of THF, 0.1 mL of HO, and 0.1 mL of MeOH were mixed and stirred at room temperature for 5 hours. The reaction mixture was extracted with HO, and the aqueous layer was adjusted to pH 5 or less by dropwise addition of 2N aqueous HCl, followed by extraction with a chloroform / IPA mixed solvent. The organic layer was dried over NaSO, and the mixture was filtered under reduced pressure and evaporated to afford intermediate 3-2'a without further purification.

[0359] Step 2: (S)-2-(3-aminoprop-1-yn-1-yl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzoic acid hydrochloride

[0360] Example 3-2' was synthesized from Intermediate 3-2'a in the same manner as in Step 2 of Preparation Example 1-1.

[0361] 1 H NMR (500 MHz, DMSO-d6) δ 10.77 (s, 1H), 8.35 (s, 3H), 8.28 (d, J = 2.1 Hz, 1H), 7.87 (dd, J = 8.6, 2.3 Hz, 1H), 7.57 (d, J = 8.5 Hz, 1H), 7.51 - 7.46 (m, 2H), 7.45 - 7.41 (m, 2H), 4.62 (d, J = 7.6 Hz, 1H), 4.03 (d, J = 5.3 Hz, 2H), 2.63 (s, 3H), 2.43 (s, 3H), 1.64 (s, 3H); MS m / z [M+H] + 573.35.

[0362] [Production Example 3-3'](S)-N-(4-(3-aminoprop-1-yn-1-yl)-3-(hydroxymethyl)phenyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0363] TIFF2025503429000267.tif40170

[0364] Step 1: (5-amino-2-iodophenyl)methanol (Intermediate 3-3a)

[0365] TIFF2025503429000268.tif12170

[0366] Methyl 5-amino-2-iodobenzoate (0.4 mmol, 1.0 eq) was dissolved in dry THF, and lithium aluminum hydride solution (2.0 M in THF, 2.0 eq) was slowly added dropwise. The mixture was then heated at 50°C for 2 hours. The pH of the reaction mixture was adjusted to 6-7 with 2N HCl aqueous solution, and the compound was extracted with EtOAc. The organic layer was dried over NaSO and filtered under reduced pressure. The mixture was distilled under reduced pressure and purified using a flash column to synthesize intermediate 3-3a.

[0367] Step 2: 3-(((tert-butyldimethylsilyl)oxy)methyl)-4-iodoaniline (Intermediate 3-3b)

[0368] TIFF2025503429000269.tif12170

[0369] Intermediate 3-3a (1.1 mmol, 1.0 eq) synthesized in Step 1 and imidazole (2.2 eq) were dissolved in dry DCM, and TBSCl (1.1 eq) was added. The gas in the reaction vessel was replaced with Ar, and the mixture was stirred at room temperature for 30 minutes. The mixture was distilled under reduced pressure and purified using a flash column to synthesize intermediate 3-3b.

[0370] Step 3: tert-butyl(3-(4-amino-2-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)prop-2-yn-1-yl)carbamate (Intermediate 3-3c)

[0371] TIFF2025503429000270.tif24170

[0372] Intermediate 3-3c was synthesized from intermediate 3-3b synthesized in step 2 in the same manner as in step 1 of Production Example 1-1.

[0373] Step 4: tert-butyl(S)-(3-(2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6 H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)phenyl)prop-2-yn-1-yl)carbamate (intermediate 3-3'a)

[0374] TIFF2025503429000271.tif39170

[0375] Intermediate 3-3'a was synthesized from Intermediate 3-3c in the same manner as in Step 1 of Production Example 1-1'.

[0376] Step 5: tert-butyl(S)-(3-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazol o[4,3-a][1,4]diazepin-6-yl)acetamido)-2-(hydroxymethyl)phenyl)prop-2-yn-1-yl)carbamate (intermediate 3-3'b)

[0377] TIFF2025503429000272.tif39170

[0378] Intermediate 3-3'a (0.2 mmol, 1.0 eq) was dissolved in 1.5 mL of THF, and then TBAF (1.0 M in THF, 1.5 eq) was added dropwise at 0° C. After 3 hours, the solvent was distilled under reduced pressure, and the mixture was purified by flash column chromatography to obtain Intermediate 3-3'b.

[0379] Step 6: (S)-N-(4-(3-aminoprop-1-yn-1-yl)-3-(hydroxymethyl)phenyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0380] Example 3-3' was synthesized using Intermediate 3-3'b in the same manner as in Step 2 of Preparation Example 1-1.

[0381] 11H NMR (400 MHz, DMSO-d6) δ 10.54 (s, 1H), 8.37 (s, 3H), 7.83 (s, 1H), 7.63 (d, J = 7.8 Hz, 1H), 7.48 (s, 2H), 7.44 (s, 2H), 7.36 (s, 1H), 4.64 (s, 3H), 4.00 (s, 2H), 3.54 (s, 2H), 2.61 (s, 3H), 2.43 (s, 3H), 1.64 (s, 3H); MS m / z [M+H] + 559.35..

[0382] [Production Example 3-8'] (S)-2-(3-aminoprop-1-yn-1-yl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzyl dihydrogen phosphate trifluoroacetic acid

[0383] TIFF2025503429000273.tif36170

[0384] Step 1: tert-butyl (S)-(3-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)-2-(((di-tert-butoxyphosphoryl)oxy)methyl)phenyl)prop-2-yn-1-yl)carbamate (Intermediate 3-8'a)

[0385] TIFF2025503429000274.tif35170

[0386] Intermediate 3-3'b (0.1 mmol, 1.0 eq) and tetrazole (10.0 eq) were dissolved in 1.7 mL of dry DMF, and the gas in the reaction vessel was replaced with Ar gas. Di-tert-butyl N,N-diisopropylphosphoramidite (5.0 eq) was added and stirred at room temperature for 3 hours. H2O2 (excess) was added dropwise at -20 °C, and after 10 minutes, the reaction was quenched with NaHSO3. The reaction mixture was extracted with EtOAc and H2O, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column to synthesize intermediate 3-8'a.

[0387] Step 2: (S)-2-(3-aminoprop-1-yn-1-yl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzyl dihydrogen phosphate trifluoroacetic acid

[0388] Example 3-8' was synthesized from Intermediate 3-8'a in the same manner as in Step 4 of Preparation Example 4-1'.

[0389] 1 H NMR (500 MHz, DMSO-d6) δ 10.56 (s, 1H), 8.56 (s, 3H), 7.72 (d, J = 2.2 Hz, 1H), 7.65 (dd, J = 8.4, 2.2 Hz, 1H), 7.47 - 7.42 (m, 2H), 7.40 - 7.34 (m, 3H), 4.90 (d, J = 5.5 Hz, 2H), 4.56 (t, J = 7.2 Hz, 1H), 3.93 (s, 2H), 3.53 - 3.45 (m, 2H), 2.56 (s, 3H), 2.38 (d, J = 0.9 Hz, 3H), 1.59 (d, J = 0.9 Hz, 3H). 31P NMR (202 MHz, DMSO-d6) δ -0.47; MS m / z [M+H] + 639.26.

[0390] [Production Example 3-25']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzoate hydrochloride

[0391] TIFF2025503429000275.tif31170

[0392] Intermediate 3-25'a (methyl(S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzoate) was synthesized from intermediate 3-25a in the same manner as in Preparation Example 1-1', and then Example 3-25' was synthesized.

[0393] 1H NMR (400 MHz, DMSO-d6) δ 10.89 (s, 1H), 8.46 (s, 3H), 8.10 (d, J = 2.2 Hz, 1H), 7.93 (d, J = 8.7 Hz, 1H), 7.67 (dd, J = 8.7, 2.2 Hz, 1H), 7.49 (d, J = 8.4 Hz, 2H), 7.42 (d, J = 8.5 Hz, 2H), 4.62 (t, J = 7.2 Hz, 1H), 4.03 (t, J = 5.6 Hz, 2H), 3.85 (s, 3H), 3.58 (d, J = 7.2 Hz, 2H), 2.63 (s, 3H), 2.43 (s, 3H), 1.64 (s, 3H); MS m / z [M+H] + 587.25.

[0394] [Production Example 3-26'](S)-2-(3-aminoprop-1-yn-1-yl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)benzoic acid hydrochloride

[0395] TIFF2025503429000276.tif31170

[0396] Intermediate 3-25'a is produced by the same method as Production Example 3-2', and Example 3-26' is synthesized by the same method.

[0397] 1H NMR (500 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.38 (s, 2H), 8.06 (d, J = 2.2 Hz, 1H), 7.88 (d, J = 8.7 Hz, 1H), 7.59 (dd, J = 8.7, 2.2 Hz, 1H), 7.46 (d, J = 8.8 Hz, 2H), 7.39 (d, J = 8.5 Hz, 2H), 4.66 - 4.47 (m, 1H), 4.05 - 3.89 (m, 2H), 3.62 - 3.45 (m, 2H), 2.59 (d, J = 1.1 Hz, 3H), 2.39 (d, J = 1.0 Hz, 3H), 1.65 - 1.52 (m, 3H); MS m / z [M+H] + 573.35.

[0398] [Production Example 3-27'](S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-(hydroxymethyl)phenyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0399] TIFF2025503429000277.tif29170

[0400] Step 1: tert-butyl(3-(5-amino-2-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)prop-2-yn-1-yl)carbamate (Intermediate 3-27c)

[0401] TIFF2025503429000278.tif24170

[0402] Intermediate 3-27c was synthesized from methyl 4-amino-2-iodobenzoate in the same manner as in steps 1 to 3 of Production Example 3-3'.

[0403] Step 2: (S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-(hydroxymethyl)phenyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0404] Intermediate 3-27'a (tert-butyl(S)-(3-(2-(((tert-butyldimethylsilyl)oxy)methyl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)phenyl)prop-2-yn-1-yl)carbamate) was synthesized from intermediate 3-27c in the same manner as in Preparation Example 3-3', and then Example 3-27' was synthesized.

[0405] 1 H NMR (400 MHz, DMSO-d6) δ 10.54 (s, 1H), 8.53 (s, 3H), 7.87 (s, 1H), 7.69 - 7.55 (m, 1H), 7.53 - 7.47 (m, 3H), 7.43 (d, J = 8.3 Hz, 2H), 4.88 (s, 1H), 4.62 (d, J = 14.8 Hz, 2H), 4.09 - 3.96 (m, 2H), 3.54 (td, J = 7.3, 6.8, 3.5 Hz, 2H), 2.65 (s, 3H), 2.43 (s, 3H), 1.64 (s, 3H); MS m / z [M+H] + 559.25.

[0406] [Production Example 3-49']N-(4-(3-amino-4-hydroxybut-1-yn-1-yl)phenyl)-2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0407] TIFF2025503429000279.tif45170

[0408] Example 3-49' was synthesized from Intermediate 3-49b in the same manner as in Preparation Example 1-1'.

[0409] MS m / z [M+H] + 559.35.

[0410] [Production Example 3-51']N-(3-(3-amino-4-hydroxybut-1-yn-1-yl)phenyl)-2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0411] TIFF2025503429000280.tif29170

[0412] Step 1: tert-butyl 4-((3-aminophenyl)ethynyl)-2,2-dimethyloxazolidine-3-carboxylate (Intermediate 3-51a)

[0413] TIFF2025503429000281.tif31170

[0414] Intermediate 3-51a was synthesized from the starting material 3-iodoaniline and intermediate 3-49a in the same manner as in Step 2 of Preparation Example 3-49.

[0415] Step 2: N-(3-(3-amino-4-hydroxybut-1-yn-1-yl)phenyl)-2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0416] Example 3-51' was synthesized from Intermediate 3-51a in the same manner as in Preparation Example 1-1'.

[0417] 1 H NMR (500 MHz, DMSO-d6) δ 10.57 (s, 1H), 8.43 (bs, 3H), 7.93 (t, J = 1.8 Hz, 1H), 7.59 (ddd, J = 8.3, 2.2, 1.1 Hz, 1H), 7.53 - 7.46 (m, 2H), 7.45 - 7.34 (m, 3H), 7.15 (dt, J = 7.8, 1.3 Hz, 1H), 4.61 (dd, J = 7.6, 6.8 Hz, 1H), 4.03 - 3.96 (m, 2H), 3.55 - 3.50 (m, 2H), 2.62 (s, 3H), 2.43 (s, 3H), 1.63 (s, 3H); MS m / z [M+H] + 559.35.

[0418] [Production Example 3-57']2-amino-4-(4-(2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)phenyl)but-3-yn-1-yl dihydrogen phosphate trifluoroacetic acid

[0419] TIFF2025503429000282.tif53170

[0420] Step 1: tert-butyl(4-(4-(2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)phenyl)-1-hydroxybut-3-yn-2-yl)carbamate (Intermediate 3-57a)

[0421] TIFF2025503429000283.tif45170

[0422] Example 3-49' (0.1 mmol, 1.0 eq) was dissolved in 3 mL of MeOH, and then TEA (excess) and BocO (excess) were added dropwise at 0 °C and stirred for 6 hours. The reaction mixture was extracted with DCM and H2O, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column to synthesize intermediate 3-57'a.

[0423] Step 2:2-amino-4-(4-(2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)phenyl)but-3-yn-1-yl dihydrogen phosphate trifluoroacetic acid

[0424] Example 3-57' was synthesized using intermediate 3-57'a in the same manner as in Preparation Example 3-8'.

[0425] 1H NMR (500 MHz, DMSO-d6) δ 10.53 (s, 1H), 7.70 - 7.63 (m, 2H), 7.49 - 7.42 (m, 2H), 7.42 - 7.36 (m, 4H), 4.54 (dd, J = 4.1, 2.4 Hz, 1H), 4.11 - 3.97 (m, 2H), 3.49 (d, J = 7.2 Hz, 2H), 2.56 (s, 3H), 2.38 (d, J = 1.0 Hz, 3H), 1.59 (d, J = 1.0 Hz, 3H). 31 P NMR (202 MHz, DMSO-d6) δ -0.60; MS m / z [M+H] + 639.36.

[0426] [Production Example 4-1'](S)-N-(3-(5-(3-aminoprop-1-yn-1-yl)furan-2-yl)prop-2-yn-1-yl)-2-(4-(4-chloroph enyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0427] TIFF2025503429000284.tif31170

[0428] Step 1: 2-bromo-5-iodofuran (Intermediate 4-1a)

[0429] TIFF2025503429000285.tif12170

[0430] 2-Bromofuran (2.0 mmol, 1.0 eq), NIS (1.1 eq), and PTSA (0.1 eq) were dissolved in 4.0 mL of EtOH and stirred at 50 °C for 10 minutes. The reaction was terminated with NaSO. The reaction mixture was extracted with EtOAc and H2O, and the organic layer was dried over NaSO and then evaporated under reduced pressure. The compound was synthesized without further purification.

[0431] Step 2: tert-butyl(3-(5-bromofuran-2-yl)prop-2-yn-1-yl)carbamate (Intermediate 4-1b)

[0432] TIFF2025503429000286.tif10170

[0433] Intermediate 4-1b was synthesized from Intermediate 4-1a in the same manner as in Step 1 of Production Example 1-1.

[0434] Step 3: tert-butyl(3-(5-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)furan-2-yl)prop-2-yn-1-yl)carbamate (intermediate 4-1c)

[0435] TIFF2025503429000287.tif31170

[0436] Intermediate 4-1c was synthesized from intermediate 4-1b in the same manner as in Step 2 of Production Example 5-116.

[0437] Step 4: 2-(3-(5-(3-aminoprop-1-yn-1-yl)furan-2-yl)prop-2-yn-1-yl)isoindoline-1,3-dione trifluoroacetic acid (Intermediate 4-1d)

[0438] TIFF2025503429000288.tif29170

[0439] Intermediate 4-1c (0.18 mmol, 1.0 eq) was dissolved in 3.0 mL of DCM, and TFA (excess) was added dropwise and stirred at room temperature for 1 hour. The reaction mixture was distilled under reduced pressure to give Intermediate 4-1d without further purification.

[0440] Step 5: (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6 -yl)-N-(3-(5-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)furan-2-yl)prop-2-yn-1-yl)acetamide (intermediate 4-1'a)

[0441] TIFF2025503429000289.tif31170

[0442] JQ1 carboxylic acid (0.2 mmol, 1.0 eq), intermediate 4-1d (1.2 eq), and HATU (1.5 eq) were dissolved in dry DCM, and the gas in the reaction vessel was replaced with Ar gas. DIPEA (3.0 eq) was added dropwise, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and purified via flash column to synthesize intermediate 4-1'a.

[0443] Step 6: (S)-N-(3-(5-(3-aminoprop-1-yn-1-yl)furan-2-yl)prop-2-yn-1-yl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0444] Example 4-1' was synthesized from Intermediate 4-1'a in the same manner as in Step 3 of Production Example 5-116 and Step 2 of Production Example 1-1.

[0445] 11H NMR (500 MHz, DMSO-d6) δ 8.83 (t, J = 5.6 Hz, 1H), 8.45 (s, 3H), 7.43 - 7.25 (m, 4H), 6.92 (d, J = 3.6 Hz, 1H), 6.86 (d, J = 3.6 Hz, 1H), 4.50 (dd, J = 8.5, 5.9 Hz, 1H), 4.27 - 4.13 (m, 2H), 4.03 (q, J = 5.6 Hz, 2H), 3.33 - 3.15 (m, 2H), 2.57 (s, 3H), 2.38 (d, J = 0.9Hz, 3H), 1.56 (d, J = 0.9 Hz, 3H); MS m / z [M+H] + 557.28.

[0446] [Production Example 4-10’] (S)-N-(3-(5-(3-aminoprop-1-yn-1-yl)thiophen-2-yl)prop-2-yn-1-yl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0447] TIFF2025503429000290.tif31170

[0448] Step 1: 2-(3-(5-(3-aminoprop-1-yn-1-yl)thiophen-2-yl)prop-2-yn-1-yl)isoindoline-1,3-dione trifluoroacetic acid (Intermediate 4-10d)

[0449] TIFF2025503429000291.tif37170

[0450] Intermediate 4-10d was synthesized from 2-bromothiophene via intermediate 4-10b (tert-butyl(3-(5-bromothiophen-2-yl)prop-2-yn-1-yl)carbamate) in the same manner as in steps 1 to 4 of Production Example 4-1′.

[0451] Step 2: (S)-N-(3-(5-(3-aminoprop-1-yn-1-yl)thiophen-2-yl)prop-2-yn-1-yl)-2-(4-(4-chloroph enyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0452] Example 4-10' was synthesized from Intermediate 4-10d in the same manner as in Step 6 of Preparation Example 4-1'.

[0453] 1 H NMR (500 MHz, DMSO-d6) δ 8.85 (s, 1H), 8.47 (s, 2H), 7.52 - 7.35 (m, 4H), 7.34 (d, J = 3.9 Hz, 1H), 7.31 (d, J = 3.8 Hz, 1H), 4.60 - 4.49 (m, 1H), 4.23 (qd, J = 18.0, 5.6 Hz, 2H), 4.10 - 4.02 (m, 2H), 3.41 - 3.28 (m, 1H), 3.27 - 3.17 (m, 1H), 2.62 (d, J = 2.9 Hz, 3H), 2.42 (d, J = 2.9 Hz, 3H), 1.61 (d, J = 5.5 Hz, 3H); MS m / z [M+H] + 573.21.

[0454] [Production Example 4-128']((S)-N-(3-(4-(3-aminoprop-1-yn-1-yl)phenyl)prop-2-yn-1-yl)-2-(4-(4-chlorophe nyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0455] TIFF2025503429000292.tif31170

[0456] Step 1: 3-(4-iodophenyl)prop-2-yn-1-amine hydrochloride (Intermediate 4-128a)

[0457] TIFF2025503429000293.tif11170

[0458] Intermediate 4-128a was synthesized from 1,4-Diiodobenzene in the same manner as in Production Example 1-1.

[0459] Step 2: (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-N-(3-(4-iodophenyl)prop-2-yn-1-yl)acetamide (Intermediate 4-128a)

[0460] TIFF2025503429000294.tif31170

[0461] Intermediate 4-128'a was synthesized from intermediate 4-128a in the same manner as in Step 5 of Preparation Example 4-1'.

[0462] Step 3: ((S)-N-(3-(4-(3-aminoprop-1-yn-1-yl)phenyl)prop-2-yn-1-yl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0463] Example 4-128' was synthesized from Intermediate 4-128'a in the same manner as in Preparation Example 1-1.

[0464] 1 H NMR (500 MHz, DMSO-d6) δ 8.82 (t, J = 5.5 Hz, 1H), 8.35 (bs, 3H), 7.51 - 7.32 (m, 8H), 4.53 (dd, J = 8.4, 5.9 Hz, 1H), 4.21 (qd, J = 17.8, MS m / z [M+H] + 567.25.

[0465] [Production Example 4-129'](S)-N-(3-(3-(3-aminoprop-1-yn-1-yl)phenyl)prop-2-yn-1-yl)-2-(4-(4-chlorophe nyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0466] TIFF2025503429000295.tif31170

[0467] Example 4-129' was synthesized from 1,3-diiodobenze in the same manner as in Preparation Example 4-128'.

[0468] 1 H NMR (500 MHz, DMSO-d6)δ 8.82 (t, J = 5.5 Hz, 1H), 8.39 (bs, 3H), 7.54 - 7.32 (m, 8H), 4.53 (dd, J = 8.4, 5.9 Hz, 1H), 4.20 (qd, J = 17.7, MS m / z [M+H] + 567.25.

[0469] [Production Example 4-147'](S)-N-(4-(5-(4-aminobut-1-yn-1-yl)furan-2-yl)but-3-yn-1-yl)-2-(4-(4-chloroph enyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0470] TIFF2025503429000296.tif36170

[0471] Step 1: tert-butyl(4-(5-bromofuran-2-yl)but-3-yn-1-yl)carbamate (Intermediate 4-147a)

[0472] TIFF2025503429000297.tif10170

[0473] Intermediate 4-147a was synthesized in the same manner as in Step 1 of Production Example 1-1 using intermediate 4-1a, tert-butyl but-3-yn-1-ylcarbamate.

[0474] Step 2: tert-butyl(4-(5-(4-(1,3-dioxoisoindolin-2-yl)but-1-yn-1-yl)furan-2-yl)but-3-yn-1-yl)carbamate (Intermediate 4-147b)

[0475] TIFF2025503429000298.tif16170

[0476] Intermediate 4-147b was synthesized in the same manner as in Step 2 of Preparation Example 5-116 using intermediate 4-147a and 2-(but-3-yn-1-yl)isoindoline-1,3-dione.

[0477] Step 3: (S)-N-(4-(5-(4-aminobut-1-yn-1-yl)furan-2-yl)but-3-yn-1-yl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0478] Example 4-147' was synthesized from Intermediate 4-147b in the same manner as in Steps 4 to 6 of Production Example 4-1'.

[0479] 1 H NMR (500 MHz, Methanol-d4) δ 7.53 - 7.47 (m, 2H), 7.43 (dd, J = 8.7, 3.0 Hz, 2H), 6.56 (d, J = 3.5 Hz, 1H), 6.50 (d, J = 3.5 Hz, 1H), 4.98 (s, 1H), 3.51 - 3.44 (m, 2H), 3.44 - 3.40 (m, 2H), 3.17 - 3.12 (m, 2H), 2.86 - 2.83 (m, 5H), 2.70 (td, J = 6.6, 3.5 Hz, 2H), 2.50 - 2.44 (m, 2H), 1.73 - 1.66 (m, 2H); MS m / z [M+H] +585.21.

[0480] [Production Example 4-177'](S)-N-(3-(5-(3-aminoprop-1-yn-1-yl)pyridin-2-yl)prop-2-yn-1-yl)-2-(4-(4-chloro phenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0481] TIFF2025503429000299.tif31170

[0482] Step 1: tert-butyl(3-(5-bromopyridin-2-yl)prop-2-yn-1-yl)carbamate (Intermediate 4-177a)

[0483] TIFF2025503429000300.tif9170

[0484] Intermediate 4-177a was synthesized using 5-Bromo-2-iodopyridine as a starting material in the same manner as in Step 1 of Production Example 1-1.

[0485] Step 2: tert-butyl(3-(5-(3-hydroxyprop-1-yn-1-yl)pyridin-2-yl)prop-2-yn-1-yl)carbamate (Intermediate 4-177b)

[0486] TIFF2025503429000301.tif9170

[0487] Intermediate 4-177a (0.7 mmol, 1.0 eq), propargyl alcohol (2.0 eq), Pd(PPh3)4 (0.1 eq), CuI (0.3 eq), TBAI (3.0 eq), TEA (0.6 mL), and dry THF (2.4 mL) were mixed and then the gas in the reaction vessel was replaced with Ar gas. The reaction was then heated at 100°C for 50 minutes in a microwave oven. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column chromatography to yield intermediate 4-177b.

[0488] Step 3: 3-(6-(3-aminoprop-1-yn-1-yl)pyridin-3-yl)prop-2-yn-1-ol hydrochloride (Intermediate 4-177c)

[0489] TIFF2025503429000302.tif9170

[0490] Intermediate 4-177c was synthesized from Intermediate 4-177b in the same manner as in Step 2 of Production Example 1-1.

[0491] Step 4: 2,5-dioxopyrrolidin-1-yl(S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetate(JQ1 NHS ester)

[0492] TIFF2025503429000303.tif31170

[0493] JQ1 carboxylic acid (1.0 mmol, 1.2 eq), N-hydroxysuccinimide (2.0 eq), EDC (2.0 eq), and 15 mL of DCM were mixed and the gas in the reaction vessel was replaced with Ar gas. The mixture was stirred overnight at room temperature. The reaction mixture was distilled under reduced pressure and purified through a flash column to synthesize the intermediate JQ1 NHS ester.

[0494] Step 5: (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]dia zepin-6-yl)-N-(3-(5-(3-hydroxyprop-1-yn-1-yl)pyridin-2-yl)prop-2-yn-1-yl)acetamide (intermediate 4-177'a)

[0495] TIFF2025503429000304.tif31170

[0496] JQ1 NHS ester (0.3 mmol, 1.0 eq) and intermediate 4-177c (1.0 eq) were dissolved in dry DCM, and the gas in the reaction vessel was replaced with Ar gas. DIPEA (2.4 eq) was added dropwise, and the mixture was stirred at room temperature overnight. The reaction mixture was distilled under reduced pressure and purified via flash column chromatography to synthesize intermediate 4-177a.

[0497] Step 6: (S)-N-(3-(5-(3-bromoprop-1-yn-1-yl)pyridin-2-yl)prop-2-yn-1-yl)-2-(4-(4-chlorophenyl)-2 ,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide (intermediate 4-177'b)

[0498] TIFF2025503429000305.tif31170

[0499] Intermediate 4-177'a (0.05 mmol, 1.0 eq) and CBr4 (2.0 eq) were dissolved in dry DCM, and the gas in the reaction vessel was replaced with Ar gas. PPh3 (2.0 eq) was slowly added, and the mixture was stirred at room temperature for 6 hours. The reaction mixture was distilled under reduced pressure and purified via flash column to synthesize intermediate 4-177'b.

[0500] Step 7: (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-y l)-N-(3-(5-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)pyridin-2-yl)prop-2-yn-1-yl)acetamide (intermediate 4-177'c)

[0501] TIFF2025503429000306.tif34170

[0502] Intermediate 4-177'b (0.02 mmol, 1.0 eq), phthalimide (1.3 eq), and CS2CO3 (2.0 eq) were dissolved in 0.6 mL of ACN and 0.2 mL of THF and stirred overnight at room temperature. The reaction mixture was distilled under reduced pressure and purified via flash column chromatography to give intermediate 4-177c'.

[0503] Step 8: (S)-N-(3-(5-(3-aminoprop-1-yn-1-yl)pyridin-2-yl)prop-2-yn-1-yl)-2-(4-(4-chlorophe nyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0504] Example 4-177' was synthesized from intermediate 4-177'c in the same manner as in Step 6 of Preparation Example 4-1'.

[0505] 1 H NMR (500 MHz, DMSO-d6) δ 8.41 (s, 3H), 7.94 (s, 1H), 7.64 (s, 1H), 7.58 (d, J = 8.1 Hz, 1H), 7.46 (d, J = 8.3 Hz, 2H), 7.40 (d, J = 8.1 Hz, 2H), 4.56 (t, J = 6.7 Hz, 1H), 4.06 - 4.00 (m, 1H), 3.86 (s, 2H), 3.75 (s, 4H), 3.53 (s, 5H), 2.57 (s, 3H), 2.38 (s, 3H), 1.59 (s, 3H); MS m / z [M+H] + 568.23.

[0506] [Production Example 4-179'](S)-N-(3-(6-(3-aminoprop-1-yn-1-yl)pyridin-3-yl)prop-2-yn-1-yl)-2-(4-(4-chloro phenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0507] TIFF2025503429000307.tif31170

[0508] Step 1: 3-(5-bromopyridin-2-yl)prop-2-yn-1-ol (Intermediate 4-179a)

[0509] TIFF2025503429000308.tif8170

[0510] 5-Bromo-2-iodopyridine (1.0 mmol, 1.0 eq), propargyl alcohol (1.1 eq), PdCl(PPh) (0.05 eq), CuI (0.1 eq), TEA (1.0 mL), and dry THF (4.0 mL) were mixed and the reaction vessel was purged with Ar gas and heated to 100 °C for 30 min under MW. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and purified via flash column chromatography to give intermediate 4-179a.

[0511] Step 2: tert-butyl(3-(6-(3-hydroxyprop-1-yn-1-yl)pyridin-3-yl)prop-2-yn-1-yl)carbamate (Intermediate 4-179b)

[0512] TIFF2025503429000309.tif9170

[0513] Intermediate 4-179b was synthesized from intermediate 4-179a and n-boc propargylamine in the same manner as in Step 2 of Preparation Example 4-177′.

[0514] Step 3: 3-(5-(3-aminoprop-1-yn-1-yl)pyridin-2-yl)prop-2-yn-1-ol hydrochloride (Intermediate 4-179c)

[0515] TIFF2025503429000310.tif9170

[0516] Intermediate 4-179c was synthesized from Intermediate 4-179b in the same manner as in Step 2 of Production Example 1-1.

[0517] ステップ4:(S)-N-(3-(6-(3-aminoprop-1-yn-1-yl)pyridin-3-yl)prop-2-yn-1-yl)-2-(4-(4-chlorophe nyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0518] Intermediate 4-179c was prepared by the same method as in Production Example 4-177' of Precision Industry 5~8, and Example 4-179' was synthesized in the same way.

[0519] 1 H NMR (500 MHz, DMSO-d6) δ 8.48 (t, J = 5.8 Hz, 1H), 8.14 (t, J = 5.7 Hz, 1H), 7.87 - 7.83 (m, 2H), 7.46 - 7.41 (m, 5H), 7.39 - 7.36 (m, 2H), 7.09 (q, J = 3.5 Hz, 2H), 4.46 (dd, J = 8.1, 6.1 Hz, 1H), 3.49 (s, 2H), 3.19 (d, J = 5.8 Hz, 3H), 3.14 - 3.09 (m, 2H), 3.05 (dq, J = 12.9, 6.2 Hz, 2H), 2.54 (d, J = 2.0 Hz, 3H), 2.35 (d, J = 3.1 Hz, 3H), 1.57 (s, 3H), 1.52 (d, J = 8.1 Hz, 2H), 1.48 (d, J = 7.6 Hz, 2H), 1.40 (s, 2H), 1.19 (s, 4H); MS m / z [M+H] + 568.23.

[0520] [Production Example 4-223'](S)-N-(3-(5-(3-(aminoxy)prop-1-yn-1-yl)thiophen-2-yl)prop-2-yn-1-yl)-2-(4-(4-chl orophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride

[0521] TIFF2025503429000311.tif31170

[0522] Step 1: tert-butyl(3-(5-(3-hydroxyprop-1-yn-1-yl)thiophen-2-yl)prop-2-yn-1-yl)carbamate (Intermediate 4-223a)

[0523] TIFF2025503429000312.tif16170

[0524] Intermediate 4-223a was synthesized from intermediate 4-10b in the same manner as in Step 2 of Preparation Example 4-177′.

[0525] Step 2: tert-butyl(3-(5-(3-((1,3-dioxoisoindolin-2-yl)oxy)prop-1-yn-1-yl)thiophen-2-yl)prop-2-yn-1-yl)carbamate (intermediate 4-223b)

[0526] TIFF2025503429000313.tif23170

[0527] PPh3 (1.2 eq) was dissolved in dry THF, then purged with Ar. Diisopropyl azodicarboxylate (1.0 eq) and intermediate 4-223a (0.2 mmol, 1.0 eq) were added at 0°C, followed by stirring for 5 minutes. N-hydroxyphthalimide (1.0 eq) was added, followed by stirring at room temperature for 5 hours. The reactant was distilled under reduced pressure and purified through a flash column to synthesize intermediate 4-223b.

[0528] Step 3: 2-((3-(5-(3-aminoprop-1-yn-1-yl)thiophen-2-yl)prop-2-yn-1-yl)oxy)isoindoline-1,3-dione trifluoroacetic acid (Intermediate 4-223c)

[0529] TIFF2025503429000314.tif18170

[0530] Intermediate 4-223c was synthesized from Intermediate 4-223b in the same manner as in Step 4 of Production Example 4-1'.

[0531] Step 4: (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)- N-(3-(5-(3-((1,3-dioxoisoindolin-2-yl)oxy)prop-1-yn-1-yl)thiophen-2-yl)prop-2-yn-1-yl)acetamide (intermediate 4-223'a)

[0532] TIFF2025503429000315.tif33170

[0533] Intermediate 4-223'a was synthesized from intermediate 4-223c in the same manner as in Step 5 of Preparation Example 4-1'.

[0534] Step 5: (S)-N-(3-(5-(3-(aminoxy)prop-1-yn-1-yl)thiophen-2-yl)prop-2-yn-1-yl)-2-(4-(4-chloro phenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide

[0535] Example 4-223' was synthesized from Intermediate 4-223'a by the same methods as in Step 3 of Preparation Example 5-116 and Step 2 of Preparation Example 1-1 in this order.

[0536] 1 H NMR (500 MHz, Methanol-d4) δ 7.50 - 7.43 (m, 2H), 7.33 - 7.26 (m, 2H), 7.22 (d, J = 3.8 Hz, 1H), 7.19 (d, J = 3.9 Hz, 1H), 4.80 - 4.74 (m, 1H), 4.49 - 4.37 (m, 2H), 4.22 (d, J = 17.8 Hz, 1H), 3.51 (dd, J = 15.1, 9.6 Hz, 1H), 3.43 - 3.32 (m, 2H), 2.79 (s, 3H), 2.47 (s, 3H), 1.70 (s, 3H); MS m / z [M+H] + 589.21.

[0537] [Production Example 5-116']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl -6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)prop-1-yn-1-yl)benzoate hydrochloride

[0538] TIFF2025503429000316.tif31170

[0539] Example 5-116' was synthesized from Intermediate 5-116c in the same manner as in Preparation Example 1-1'.

[0540] 1 H NMR (500 MHz, DMSO-d6) δ 8.81 (t, J = 5.4 Hz, 1H), 8.38 (s, 3H), 7.88 (d, J = 8.2 Hz, 1H), 7.59 (d, J = 1.7 Hz, 1H), 7.55 (dd, J = 8.2, 1.7 Hz, 1H), 7.40 - 7.36 (m, 2H), 7.32 (d, J = 8.9 Hz, 2H), 4.50 (dd, J = 8.3, 6.2 Hz, 1H), 4.19 (t, J = 5.5 Hz, 2H), 4.00 (q, J = 5.7 Hz, 2H), 3.84 (s, 3H), 3.37 - 3.17 (m, 2H), 2.56 (s, 3H), 2.37 (d, J = 0.9 Hz, 3H), 1.56 (d, J = 0.9 Hz, 3H); MS m / z [M+H] + 625.37.

[0541] [Manufacturing example II'] A TBM-UBM compound in which a compound (JQ1) exhibiting activity as a BRD4 (bromodomain-containing protein 4) inhibitor is linked to a compound corresponding to Formula II of the present invention was synthesized as follows.

[0542] [Production Example 4-124'](S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanamide hydrochloride

[0543] TIFF2025503429000317.tif45170

[0544] Step 1: (S)-3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanoic acid (intermediate 4-124'a)

[0545] TIFF2025503429000318.tif31170

[0546] Intermediate 4-124'a was synthesized from β-alanine in the same manner as in Step 5 of Preparation Example 4-177'.

[0547] Step 2: (S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanamide hydrochloride

[0548] Example 4-124' was synthesized from Intermediate 4-124'a and Intermediate 1-1a in the same manner as in Preparation Example 1-1'.

[0549] 1 H NMR (500 MHz, DMSO-d6) δ 10.29 (s, 1H), 8.38 (s, 3H), 7.73 - 7.60 (m, 2H), 7.52 (d, J = 8.6 Hz, 1H), 7.38 (tt, J = 8.7, 4.2 Hz, 5H), 4.51 (d, J = 6.9 Hz, 1H), 3.92 (d, J = 5.8 Hz, 2H), 3.40 - 3.22 (m, 4H), 3.30 - 3.00 (m, 2H), 2.63 - 2.57 (m, 3H), 2.56 - 2.50 (m, 2H), 2.38 (s, 3H), 1.60 - 1.54 (m, 3H); MS m / z [M+H] + 600.32.

[0550] [Production Example 4-125'](S)-N-(4-(3-aminoprop-1-yn-1-yl)-3-(hydroxymethyl)phenyl)-3-(2-(4-(4-chlorophenyl)- 2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanamide hydrochloride

[0551] TIFF2025503429000319.tif45170

[0552] Step 1: tert-butyl(S)-(3-(2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thi eno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanamido)phenyl)prop-2-yn-1-yl)carbamate (intermediate 4-125'a)

[0553] TIFF2025503429000320.tif46170

[0554] Intermediate 4-125'a was synthesized from Intermediate 4-124'a and Intermediate 3-3c in the same manner as in Step 1 of Preparation Example 1-1'.

[0555] Step 2: (S)-N-(4-(3-aminoprop-1-yn-1-yl)-3-(hydroxymethyl)phenyl)-3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanamide hydrochloride

[0556] TIFF2025503429000321.tif48170

[0557] Example 4-125 was synthesized from Intermediate 4-125'a in the same manner as in Steps 5 and 6 of Production Example 3-3'.

[0558] 1 H NMR (500 MHz, DMSO-d6) δ 10.24 (d, J = 11.4 Hz, 1H), 8.41 (d, J = 35.7 Hz, 3H), 7.76 (s, 1H), 7.63 - 7.57 (m, 1H), 7.42 - 7.35 (m, 4H), 7.29 (d, J = 8.3 Hz, 1H), 4.58 (s, 1H), 4.56 - 4.42 (m, 2H), 3.94 (d, J = 6.3 Hz, 2H), 3.42 - 3.30 (m, 2H), 3.29 - 3.10 (m, 2H), 2.59 (q, J = 4.9, 4.2 Hz, 3H), 2.53 (t, J = 7.0 Hz, 2H), 2.38 (s, 3H), 1.61 - 1.55 (m, 3H); MS m / z [M+H] + 630.46.

[0559] [Production Example 4-126']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-5-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanamido)benzoate hydrochloride

[0560] TIFF2025503429000322.tif39170

[0561] Example 4-126' was synthesized from Intermediate 4-124'a and Intermediate 3-1a in the same manner as in Preparation Example 1-1'.

[0562] 1 1H NMR (500 MHz, DMSO-d6) δ 10.49 (d, J = 13.2 Hz, 1H), 8.39 (d, J = 25.3 Hz, 3H), 8.24 (d, J = 2.1 Hz, 1H), 7.83 (dd, J = 8.5, 2.3 Hz, 1H), 7.50 (d, J = 8.5 Hz, 1H), 7.43 - 7.33 (m, 4H), 4.50 (d, J = 5.6 Hz, 1H), 4.01 - 3.91 (m, 2H), 3.83 (s, 3H), 3.43 - 3.30 (m, 2H), 3.27 - 3.10 (m, 2H), 2.58 (dd, J = 3.9, 1.6 Hz, 3H), 2.54 (d, J = 6.8 Hz, 2H), 2.41 - 2.34 (m, 3H), 1.58 - 1.53 (m, 3H); MS m / z [M+H] + 658.47.

[0563] [Production Example 4-148'] (S)-5-(3-aminoprop-1-yn-1-yl)-N-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)but-2-yn-1-yl)furan-2-carboxamide hydrochloride

[0564] TIFF2025503429000323.tif36170

[0565] Step 1: tert-butyl(4-aminobut-2-yn-1-yl)carbamate (Intermediate 4-148a)

[0566] TIFF2025503429000324.tif13170

[0567] Intermediate 4-148a was synthesized from But-2-yne-1,4-diamine in the same manner as in Step 1 of Preparation Example 3-57'.

[0568] Step 2: (S)-N-(4-aminobut-2-yn-1-yl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride (Intermediate 4-148a)

[0569] TIFF2025503429000325.tif31170

[0570] JQ1 Intermediate 4-148'a was synthesized from carboxylic acid and intermediate 4-148a in the same manner as in Production Example 1-1'.

[0571] Step 3: (S)-5-(3-aminoprop-1-yn-1-yl)-N-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[ 3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)but-2-yn-1-yl)furan-2-carboxamide hydrochloride

[0572] Example 4-148' was synthesized from Intermediate 4-148'a and Intermediate 4-227b in the same manner as in Preparation Example 1-1'.

[0573] 1H NMR (500 MHz, DMSO-d6) δ 8.96 (t, J = 5.7 Hz, 1H), 8.61 (t, J = 5.5 Hz, 1H), 8.41 (s, 3H), 7.45 (d, J = 8.7 Hz, 2H), 7.41 - 7.35 (m, 2H), 7.21 (d, J = 3.6 Hz, 1H), 6.97 (d, J = 3.6 Hz, 1H), 4.47 (dd, J = 8.2, 6.0 Hz, 1H), 4.08 - 3.97 (m, 3H), 3.28 - 3.12 (m, 2H), 2.56 (s, 3H), 2.37 (s, 3H), 1.58 (s, 3H). MS m / z [M+H] + 614.25.

[0574] [Production Example 4-149']((S)-5-(3-aminoprop-1-yn-1-yl)-N-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H- thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butyl)furan-2-carboxamide hydrochloride

[0575] TIFF2025503429000326.tif34170

[0576] Example 4-149' was synthesized from tert-butyl(4-aminobutyl)carbamate in the same manner as in Steps 2 and 3 of Preparation Example 4-148'.

[0577] 1H NMR (500 MHz, DMSO-d6) δ 8.57 (t, J = 5.8 Hz, 1H), 8.50 (s, 3H), 8.24 (t, J = 5.6 Hz, 1H), 7.49 (d, J = 8.8 Hz, 2H), 7.41 (d, J = 8.7 Hz, 2H), 7.18 (d, J = 3.6 Hz, 1H), 6.99 (d, J = 3.6 Hz, 1H), 4.52 (dd, J = 8.3, 6.0 Hz, 1H), 4.08 (s, 2H), 3.26 - 3.18 (m, 4H), 3.17 - 3.08 (m, 2H), 2.61 (s, 3H), 2.41 (d, J = 0.9 Hz, 3H), 1.61 (d, J = 0.9 Hz, 3H), 1.52 (d, J = 7.0 Hz, 2H), 1.47 (d, J = 7.1 Hz, 2H); + 618.16.

[0578] [Production Example 4-161'](S)-N-(3-(3-aminoprop-1-yn-1-yl)phenyl)-3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanamide hydrochloride

[0579] TIFF2025503429000327.tif37170

[0580] Intermediate 4-124'a and Intermediate 1-3a are synthesized by the same method as Production Example 1-1' and Example 4-161'.

[0581] MS m / z [M+H] + 600.36.

[0582] [Production Example 4-162'](S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide hydrochloride

[0583] TIFF2025503429000328.tif36170

[0584] Step 1: (S)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanoic acid (Intermediate 4-162a)

[0585] TIFF2025503429000329.tif31170

[0586] Intermediate 4-162'a was synthesized from 4-Aminobutanoic acid in the same manner as in Step 5 of Preparation Example 4-177'.

[0587] Step 2: (S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide hydrochloride

[0588] Example 4-162' was synthesized from Intermediate 4-162'a and Intermediate 1-1a in the same manner as in Preparation Example 1-1'.

[0589] 11H NMR (500 MHz, DMSO-d6) δ 10.19 (s, 1H), 8.33 (s, 3H), 8.25 (d, J = 5.7 Hz, 1H), 7.66 - 7.60 (m, 2H), 7.42 (d, J = 8.9 Hz, 2H), 7.39 - 7.32 (m, 4H), 4.48 (dd, J = 8.2, 6.1 Hz, 1H), 3.93 (q, J = 5.6 Hz, 2H), 3.27 - 3.05 (m, 4H), 2.57 (s, 3H), 2.39 - 2.36 (m, 3H), 2.36 - 2.30 (m, 2H), 1.74 (q, J = 7.3 Hz, 2H), 1.59 - 1.54 (m, 3H); MS m / z [M+H] + 614.36.

[0590] [Production Example 4-163'] (S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)pentanamide hydrochloride

[0591] TIFF2025503429000330.tif48170

[0592] Step 1: (S)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)pentanoic acid (Intermediate 4-163'a)

[0593] TIFF2025503429000331.tif36170

[0594] Intermediate 4-163'a was synthesized from 5-Aminopentanoic acid in the same manner as in Step 5 of Preparation Example 4-177'.

[0595] Step 2: (S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)pentanamide hydrochloride

[0596] Example 4-163' was synthesized from Intermediate 4-163'a and Intermediate 1-1a in the same manner as in Preparation Example 1-1'.

[0597] 1 H NMR (500 MHz, DMSO-d6) δ 10.24 (d, J = 2.3 Hz, 1H), 8.42 (s, 3H), 8.24 (t, J = 5.8 Hz, 1H), 7.64 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 8.0 Hz, 2H), 7.39 - 7.29 (m, 4H), 4.51 (d, J = 6.7 Hz, 1H), 3.95 - 3.86 (m, 2H), 3.27 - 3.01 (m, 4H), 2.63 - 2.54 (m, 3H), 2.41 - 2.36 (m, 3H), 2.33 (t, J = 7.4 Hz, MS m / z [M+H] + 628.46.

[0598] [Production Example 4-164'](S)-N-(3-(3-aminoprop-1-yn-1-yl)phenyl)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)pentanamide hydrochloride

[0599] TIFF2025503429000332.tif47170

[0600] Intermediate 4-163'a and Intermediate 1-3a are synthesized by the same method as Production Example 1-1' and Example 4-164'.

[0601] 1 H NMR (500 MHz, DMSO-d6) δ 10.16 (d, J = 3.5 Hz, 1H), 8.40 (s, 3H), 8.24 (t, J = 5.8 Hz, 1H), 7.90 (d, J = 1.9 Hz, 1H), 7.49 (dd, J = 8.3, 1.9 Hz, 1H), 7.46 - 7.41 (m, 2H), 7.40 - 7.34 (m, 2H), 7.28 (t, J = 7.9 Hz, 1H), 7.07 (dt, J = 7.7, 1.3 Hz, 1H), 4.49 (dd, J = 8.3, 5.7 Hz, 1H), 3.97 - 3.85 (m, 2H), 3.26 - 2.98 (m, 4H), 2.58 (d, J = 1.1 Hz, 3H), 2.37 (d, J = 0.9 Hz, 3H), 2.31 (q, J = 8.5, 7.9 Hz, 2H), 1.58 (s, 2H), 1.56 (d, J = 0.9 Hz, 3H), 1.44 (p, J = 7.1 Hz, 2H); MS m / z [M+H] + 628.46.

[0602] [Production Example 4-165'](S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-6-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)hexanamide hydrochloride

[0603] TIFF2025503429000333.tif31170

[0604] Step 1: (S)-6-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)hexanoic acid (Intermediate 4-165a)

[0605] TIFF2025503429000334.tif34170

[0606] Intermediate 4-165'a was synthesized from 6-aminohexanoic acid in the same manner as in Step 5 of Preparation Example 4-177'.

[0607] Step 2: (S)-N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-6-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)hexanamide hydrochloride

[0608] Example 4-165' was synthesized from Intermediate 4-165'a and Intermediate 1-1a in the same manner as in Preparation Example 1-1'.

[0609] 1H NMR (500 MHz, DMSO-d6) δ 10.10 (s, 1H), 8.25 (s, 3H), 8.17 (s, 1H), 7.66 - 7.58 (m, 2H), 7.50 - 7.42 (m, 2H), 7.40 - 7.29 (m, 4H), 4.49 - 4.42 (m, 1H), 3.94 (d, J = 5.7 Hz, 2H), 3.24 - 3.02 (m, 4H), 2.55 (d, J = 3.2 Hz, 3H), 2.37 (s, 3H), 2.34 - 2.25 (m, 2H), 1.57 (s, 3H), 1.56 (d, J = 10.1 MS m / z [M+H] + 642.46.

[0610] [Production Example 4-166']((S)-5-(3-aminoprop-1-yn-1-yl)-N-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H- thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)furan-2-carboxamide hydrochloride

[0611] TIFF2025503429000335.tif34170

[0612] Step 2 of Preparation Example 5-77' and Step 2 of Preparation Example 1-1 were applied sequentially from tert-Butyl(3-aminopropyl)carbamate to obtain intermediate 4-166'a ((S)-N-(3-aminopropyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride), and then Example 4-166' was synthesized from this intermediate 4-166'a and 4-227b in the same manner as in Preparation Example 1-1'.

[0613] 1 H NMR (500 MHz, DMSO-d6) δ 8.57 (t, J = 5.8 Hz, 1H), 8.52 (s, 3H), 8.31 (t, J = 5.7 Hz, 1H), 7.48 (d, J = 8.8 Hz, 2H), 7.42 (d, J = 8.5 Hz, 2H), 7.19 (d, J = 3.6 Hz, 1H), 7.00 (d, J = 3.6 Hz, 1H), 4.53 (dd, J = 8.2, 6.1 Hz, 1H), 4.08 (t, J = 5.7 Hz, 2H), 3.29 - 3.20 (m, 4H), 3.19 - 3.11 (m, 2H), 2.61 (s, 3H), 2.41 (d, J = 0.9 Hz, 3H), 1.68 (t, J = 7.0 Hz, 2H), 1.63 - 1.60 (m, 3H); MS m / z [M+H] + 604.16.

[0614] [Production Example 4-167'](S)-5-(3-aminoprop-1-yn-1-yl)-N-(5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)pentyl)furan-2-carboxamide hydrochloride

[0615] TIFF2025503429000336.tif26170

[0616] Tert-butyl (5-aminopentyl) carbamate is produced by the same method as in Production Example 4-166 and Synthesis in Example 4-167.

[0617] 1H NMR (500 MHz, DMSO-d6) δ 8.53 (dd, J = 10.3, 4.4 Hz, 4H), 8.24 (t, J = 5.5 Hz, 1H), 7.50 (d, J = 8.8 Hz, 2H), 7.42 (d, J = 8.5 Hz, 2H), 7.18 (d, J = 3.6 Hz, 1H), 6.98 (d, J = 3.6 Hz, 1H), 4.54 (dd, J = 8.2, 6.0 Hz, 2H), 4.07 (d, J = 5.6 Hz, 2H), 3.25 - 3.17 (m, 4H), 3.11 (dt, J = 30.0, 6.9 Hz, 2H), 2.61 (s, 3H), 2.41 (d, J = 1.0 Hz, 3H), 1.63 - 1.60 (m, 4H), 1.52 (q, J = 7.3 Hz, 2H), 1.46 (q, J = 7.4 Hz, 2H), 1.32 (q, J = 8.6, 8.1 Hz, 2H); MS m / z [M+H] + 632.27.

[0618] [Synthesis 4-225'](S)-5-(3-aminoprop-1-yn-1-yl)-N-(9-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H- thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)nonyl)furan-2-carboxamide hydrochloride

[0619] TIFF2025503429000337.tif37170

[0620] tert-butyl(9-aminononyl)carbamate has a range of 4-166' and a range of 4-225'.

[0621] 1H NMR (500 MHz, DMSO-d6) δ 8.53 - 8.44 (m, 3H), 7.45 (d, J = 8.7 Hz, 2H), 7.40 - 7.37 (m, 2H), 7.36 (d, J = 8.7 Hz, 1H), 7.13 (d, J = 3.6 Hz, 1H), 7.10 (d, J = 3.1 Hz, 1H), 6.94 (d, J = 3.6 Hz, 1H), 4.49 (d, J = 2.4 Hz, 1H), 4.04 (p, J = 5.9 Hz, 2H), 3.34 - 3.30 (m, 2H), 3.17 (t, J = 9.3 Hz, 6=4H), MS m / z [M+H] + 688.42.

[0622] [Production Example 4-228'](methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoate hydrochloride

[0623] TIFF2025503429000338.tif31170

[0624] Intermediate 3-25a is produced by the same method as Production Example 5-77' and Example 4-228' is produced by the same method.

[0625] 1H NMR (500 MHz, DMSO-d6) δ 10.50 (s, 1H), 8.40 (s, 3H), 8.29 (t, J = 5.6 Hz, 1H), 8.09 (d, J = 2.1 Hz, 1H), 7.85 (d, J = 8.7 Hz, 1H), 7.55 (dd, J = 8.7, 2.2 Hz, 1H), 7.43 - 7.36 (m, 4H), 4.49 (dd, J = 7.9, 6.4 Hz, 1H), 4.01 (q, J = 5.6 Hz, 2H), 3.79 (s, 3H), 3.53 (d, J = 4.6 Hz, 1H), 3.34 (q, J = 7.0 MS m / z [M+H] + 672.34.

[0626] [Production Example 4-235']((S)-5-(3-aminoprop-1-yn-1-yl)-N-(8-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H- thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)octyl)furan-2-carboxamide hydrochloride

[0627] TIFF2025503429000339.tif45170

[0628] Example 4-235' was synthesized from tert-butyl(8-aminooctyl)carbamate in the same manner as in Preparation Example 4-166'.

[0629] 11H NMR (500 MHz, DMSO-d6) δ 8.53 - 8.44 (m, 4H), 8.16 (t, J = 5.7 Hz, 1H), 7.47 - 7.43 (m, 2H), 7.41 - 7.36 (m, 2H), 7.13 (d, J = 3.6 Hz, 1H), 6.94 (d, J = 3.6 Hz, 1H), 4.48 (dd, J = 8.2, 6.0 Hz, 1H), 4.04 (q, J = 5.7 Hz, 2H), 3.37 - 3.17 (m, 2H), 3.17 - 2.99 (m, 4H), 2.57 (d, J = 1.1 Hz, 3H), 2.40 - 2.36 (m, 3H), 1.59 - 1.56 (m, 3H), 1.40 (dd, J = 15.3, 8.4 Hz, 4H), 1.28 - 1.16 (m, 8H); MS m / z [M+H] + 674.42.

[0630] [Production Example 5-1'] ethyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoate hydrochloride

[0631] TIFF2025503429000340.tif31170

[0632] Step 1: ethyl (S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoate (Intermediate 5-1'a)

[0633] TIFF2025503429000341.tif34170

[0634] Intermediate 5-53'a (0.02 mmol, 1.0 eq) was dissolved in 0.5 mL of dry DMF, K2CO3 (3.0 eq), and the gas in the reaction vessel was replaced with Ar gas. The reaction mixture was stirred at room temperature for 30 minutes, after which iodoethane (3.0 eq) was added dropwise and the mixture was stirred at room temperature overnight. The reaction mixture was purified by flash column chromatography to synthesize intermediate 5-1'a.

[0635] Step 2: ethyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoate hydrochloride

[0636] Example 5-1' was synthesized from Intermediate 5-1'a in the same manner as in Step 2 of Preparation Example 1-1.

[0637] 1H NMR (500 MHz, DMSO-d6) δ 10.44 (s, 1H), 8.37 (s, 3H), 8.30 (t, J = 5.7 Hz, 1H), 8.17 (d, J = 2.1 Hz, 1H), 7.89 (d, J = 8.7 Hz, 1H), 7.53 (dd, J = 8.7, 2.3 Hz, 1H), 7.45 (d, J = 8.9 Hz, 2H), 7.40 (d, J = 8.7 Hz, 2H), 4.50 (dd, J = 7.9, 6.3 Hz, 1H), 4.29 (q, J = 7.1 Hz, 2H), 4.04 (q, J = 5.6 Hz, 2H), 3.23 - 3.13 (m, 4H), 2.59 (s, 3H), 2.43 (dd, J = 7.7, 2.3 Hz, 2H), 2.41 (s, 3H), 1.78 (q, J = 7.3 Hz, 2H), 1.61 (s, 3H), 1.32 (d, J = 7.2 Hz, 3H); MS m / z [M+H] + 686.48.

[0638] [Production Example 5-2']isopropyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoate hydrochloride

[0639] TIFF2025503429000342.tif31170

[0640] Example 5-2' was synthesized from intermediate 5-53'a and 2-iodopropane in the same manner as in Example 5-1'.

[0641] 1H NMR (500 MHz, DMSO-d6) δ 10.44 (d, J = 2.3 Hz, 1H), 8.37 (s, 3H), 8.30 (s, 1H), 8.17 (t, J = 2.3 Hz, 1H), 7.86 (dd, J = 8.7, 2.3 Hz, 1H), 7.52 (d, J = 8.6 Hz, 1H), 7.44 (dd, J = 8.8, 2.3 Hz, 2H), 7.40 (dd, J = 8.6, 2.3 Hz, 2H), 5.09 (ddd, J = 12.6, 6.3, 2.4 Hz, 1H), 4.54 - 4.43 (m, 1H), 4.06 - 4.01 (m, 2H), 3.24 - 3.15 (m, 4H), 2.59 (d, J = 2.3 Hz, 3H), 2.42 (s, 2H), 2.41 (d, J = 2.3 Hz, 3H), 1.79 - 1.74 (m, 2H), 1.61 (d, J = 2.3 Hz, 3H), 1.31 (dd, J = 6.3, 2.3 Hz, 6H); MS m / z [M+H] + 700.39.

[0642] [Production Example 5-4']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanamido)benzoate hydrochloride

[0643] TIFF2025503429000343.tif45170

[0644] Intermediate 4-124'a and Intermediate 3-25a are synthesized by the same method as Production Example 1-1' and Example 5-4'.

[0645] 1H NMR (500 MHz, DMSO-d6) δ 10.62 (s, 1H), 8.54 - 8.35 (m, 4H), 8.14 (d, J = 2.2 Hz, 1H), 7.90 (d, J = 8.7 Hz, 1H), 7.63 (dd, J = 8.7, 2.2 Hz, 1H), 7.47 - 7.37 (m, 4H), 4.53 (dd, J = 8.1, 6.0 Hz, 1H), 4.05 (q, J = 5.6 Hz, 2H), 3.84 (s, 3H), 3.41 (dd, J = 10.0, 6.1 Hz, 2H), 3.29 - 3.16 (m, 2H), 2.61 (s, 5H), 2.42 (d, J = 0.9 Hz, 3H), 1.61 (s, 3H); MS m / z [M+H] + 658.38.

[0646] [Production Example 5-5']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)pentanamido)benzoate hydrochloride

[0647] TIFF2025503429000344.tif47170

[0648] Example 5-5' was synthesized from intermediate 3-25a in the same manner as in Step 2 of Preparation Example 5-77'.

[0649] 1H NMR (500 MHz, DMSO-d6) δ 10.39 (s, 1H), 8.33 (s, 3H), 8.25 (t, J = 5.6 Hz, 1H), 8.15 (d, J = 2.2 Hz, 1H), 7.89 (d, J = 8.7 Hz, 1H), 7.53 (dd, J = 8.7, 2.2 Hz, 1H), 7.46 (d, J = 8.8 Hz, 2H), 7.40 (d, J = 8.6 Hz, 2H), 4.50 (dd, J = 8.4, 5.9 Hz, 1H), 4.06 (q, J = 5.7 Hz, 2H), 3.83 (s, 3H), 3.26 (dd, J = 15.0, 8.4 Hz, 2H), 3.17 (dd, J = 10.5, 4.4 Hz, 2H), 2.59 (d, J = 2.8 Hz, 3H), 2.41 - 2.40 (m, 3H), 2.39 - 2.36 (m, 2H), 1.63 (d, J = 8.2 Hz, 2H), 1.60 (d, J = 0.8 Hz, 3H), 1.51 - 1.47 (m, 2H); MS m / z [M+H] + 686.47.

[0650] [Production Example 5-6']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(6-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)hexanaido)benzoate hydrochloride

[0651] TIFF2025503429000345.tif45170

[0652] Intermediate 4-165'a and Intermediate 3-25a were synthesized by the same method as Production Example 5-77' and Example 5-6'.

[0653] 1H NMR (500 MHz, DMSO-d6) δ 10.36 (s, 1H), 8.32 (s, 3H), 8.21 (t, J = 5.7 Hz, 1H), 8.16 (d, J = 2.2 Hz, 1H), 7.89 (d, J = 8.6 Hz, 1H), 7.52 (dd, J = 8.7, 2.2 Hz, 1H), 7.49 (d, J = 8.7 Hz, 2H), 7.42 (d, J = 8.5 Hz, 2H), 4.50 (dd, J = 8.1, 6.1 Hz, 1H), 4.05 (dd, J = 11.2, 5.6 Hz, 2H), 3.83 (s, 3H), 3.25 - 3.18 (m, 2H), 2.67 - 2.62 (m, 2H), 2.59 (s, 3H), 2.41 (s, 3H), 2.38 - 2.33 (m, 4H), 1.62 (s, 3H), 1.46 (q, J = 7.4 Hz, 2H), 1.35 (d, J = 7.1 Hz, 2H); MS m / z [M+H] + 700.47.

[0654] [Production Example 5-8'](S)-N-(4-(3-aminoprop-1-yn-1-yl)-3-(hydroxymethyl)phenyl)-4-(2-(4-(4-chlorophenyl)- 2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide hydrochloride

[0655] TIFF2025503429000346.tif31170

[0656] Intermediate 3-3c and 4-162'a were synthesized using the same method as in Production Example 4-125' and Example 5-8'.

[0657] 1H NMR (500 MHz, DMSO-d6) δ 10.15 (s, 1H), 8.30 (d, J = 5.3 Hz, 4H), 7.76 (d, J = 2.2 Hz, 1H), 7.63 (dd, J = 8.4, 2.2 Hz, 1H), 7.51 - 7.42 (m, 2H), 7.41 (d, J = 8.7 Hz, 2H), 7.33 (d, J = 8.4 Hz, 1H), 4.77 (s, 1H), 4.62 (s, 2H), 4.51 (dd, J = 8.1, 6.2 Hz, 1H), 4.01 (q, J = 5.6 Hz, 2H), 3.20 - 3.08 (m, MS m / z [M+H] + 644.45.

[0658] [Production Example 5-9'](S)-N-(4-(3-aminoprop-1-yn-1-yl)-3-(hydroxymethyl)phenyl)-5-(2-(4-(4-chlorophenyl)-2 ,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)pentanamide hydrochloride

[0659] TIFF2025503429000347.tif53170

[0660] Intermediate 4-163'a and Intermediate 3-3c are synthesized by the same method as Production Example 4-125' and Example 5-9'.

[0661] 11H NMR (500 MHz, DMSO-d6) δ 10.10 (s, 1H), 8.29 (s, 3H), 8.21 (t, J = 5.6 Hz, 1H), 7.70 (d, J = 2.2 Hz, 1H), 7.60 (dd, J = 8.4, 2.2 Hz, 1H), 7.43 (d, J = 8.8 Hz, 2H), 7.37 (d, J = 8.5 Hz, 2H), 7.28 (d, J = 8.4 Hz, 1H), 4.73 (s, 1H), 4.57 (s, 2H), 4.47 (dd, J = 8.4, 5.8 Hz, 1H), 3.96 (q, J = 5.7 Hz, 2H), 3.22 (dd, J = 15.0, 8.5 Hz, 2H), 3.13 (dd, J = 10.5, 4.5 Hz, 2H), 2.55 (s, 3H), 2.38 - 2.35 (m, 3H), 2.34 - 2.29 (m, 2H), 1.56 (d, J = 0.9 Hz, 3H), 1.48 - 1.39 (m, 4H); MS m / z [M+H] + 658.34.

[0662] [Production Example 5-12’] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butoxy)benzoate hydrochloride

[0663] TIFF2025503429000348.tif39170

[0664] Step 1: methyl 4-hydroxy-2-iodobenzoate (Intermediate 5-12a)

[0665] TIFF2025503429000349.tif15170[[ID=!9]]

[0666] To a mixture of methyl 4-amino-2-iodobenzoate (1.0 mmol, 1.0 eq), 0.3 mL of H2SO4, and 3.0 mL of H2O, 1.0 eq of Na2NO3 dissolved in a small amount of H2O was added dropwise while stirring at 0 °C. After 30 minutes, the reaction mixture was filtered and the filtrate was stirred at reflux for 30 minutes. The reaction temperature was then lowered to room temperature and stirred for 18 hours. The reaction mixture was extracted with Et2O, and the organic layer was dried over MgSO4, filtered under reduced pressure, and evaporated under reduced pressure to give intermediate 5-12a without further purification.

[0667] Step 2: methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-hydroxybenzoate (Intermediate 5-12b)

[0668] TIFF2025503429000350.tif26170

[0669] Intermediate 5-12b was synthesized from Intermediate 5-12a in the same manner as in Step 1 of Production Example 1-1.

[0670] Step 3: (S)-N-(4-bromobutyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide (Intermediate 5-12'a)

[0671] TIFF2025503429000351.tif31170

[0672] Intermediate 5-12'a was synthesized from 4-Bromobutan-1-amine hydrochloride in the same manner as in Step 5 of Preparation Example 4-1'.

[0673] Step 4: methyl(S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-tr imethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butoxy)benzoate (intermediate 5-12'b)

[0674] TIFF2025503429000352.tif31170

[0675] Dry DMF was added dropwise to intermediate 5-12'a (0.03 mmol, 1.0 eq), intermediate 5-12b (4.8 eq), K2CO3 (4.8 eq), and TBAI (0.1 eq). The gas in the reaction vessel was replaced with Ar gas and the reaction was carried out at 100 °C. After 2 hours, H2O was added to the reaction mixture to terminate the reaction. The compound was extracted into the organic layer using DCM, and the organic layer was dried over MgSO4. After vacuum filtration and distillation, the mixture was purified using a flash column to synthesize intermediate 5-12'b.

[0676] Step 5: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butoxy)benzoate hydrochloride

[0677] Example 5-12' was synthesized from Intermediate 5-12'b in the same manner as in Step 2 of Preparation Example 1-1.

[0678] 11H NMR (500 MHz, DMSO-d6) δ 8.35 (s, 3H), 8.27 (t, J = 5.8 Hz, 1H), 7.89 (d, J = 8.6 Hz, 1H), 7.43 (q, J = 8.8 Hz, 4H), 7.11 - 7.06 (m, 2H), 4.50 (dd, J = 8.3, 6.0 Hz, 1H), 4.11 - 4.00 (m, 4H), 3.82 (s, 3H), 3.28 - 3.14 (m, 4H), 2.59 (s, 3H), 2.41 (s, 3H), 2.03 - 1.95 (m, 2H), 1.78 (q, J = 7.3 Hz, 2H), 1.63 - 1.56 (m, 3H); MS m / z [M+H] + 659.54.

[0679] [Production Example 5-18'] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-((3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)carbamoyl)benzoate hydrochloride

[0680] TIFF2025503.jpg34170

[0681] Step 1: methyl (S)-2-bromo-4-((3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)carbamoyl)benzoate (Intermediate 5-18'a)

[0682] TIFF2025503.jpg34170

[0683] Intermediate 5-18'a was synthesized from intermediate 4-166'a and 3-bromo-4-(methoxycarbonyl)benzoic acid in the same manner as in Step 5 of Preparation 4-1'.

[0684] Step 2: methyl(S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-((3-(2-(4-(4-chlorophenyl)-2,3,9-trimet hyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)carbamoyl)benzoate (intermediate 5-18'b)

[0685] TIFF2025503429000355.tif34170

[0686] Intermediate 5-18'b was synthesized from intermediate 5-18'a in the same manner as in Step 2 of Preparation Example 4-179'.

[0687] Step 3: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-((3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)carbamoyl)benzoate hydrochloride

[0688] Example 5-18' was synthesized from Intermediate 5-18'b in the same manner as in Step 2 of Preparation Example 1-1.

[0689] 11H NMR (500 MHz, DMSO-d6) δ 8.80 (t, J = 5.6 Hz, 1H), 8.42 (s, 3H), 8.30 (t, J = 5.7 Hz, 1H), 8.07 (t, J = 1.2 Hz, 1H), 7.99 - 7.97 (m, 2H), 7.49 - 7.39 (m, 4H), 4.51 (dd, J = 8.1, 6.1 Hz, 1H), 4.06 (q, J = 5.7 Hz, 2H), 3.89 (s, 3H), 3.38 - 3.10 (m, 6H), 2.60 (s, 3H), 2.41 (s, 3H), 1.72 (p, J = 6.9 Hz, 2H), 1.60 (s, 3H); MS m / z [M+H] + 672.42.

[0690] [Production Example 5-35'] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-((4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)cyclohexyl)amino)benzoate trifluoroacetic acid

[0691] TIFF2025503429000356.tif39170

[0692] Step 1: methyl 4-bromo-2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoate (Intermediate 5-35a)

[0693] TIFF2025503429000357.tif25170

[0694] Methyl 4-bromo-2-iodobenzoate (2.0 mmol, 1.0 eq), n-propargylphthalimide (1.1 eq), Pd(PPh3)4 (0.01 eq), CuI (0.05 eq), and 7.0 mL of ACN were mixed, and the gas in the reaction vessel was replaced with Ar gas. The reaction mixture was stirred at room temperature for 3 hours, then filtered through Celite under reduced pressure and evaporated under reduced pressure. The mixture was purified by flash column chromatography to synthesize intermediate 5-35a.

[0695] Step 2: methyl 4-((4-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoate (intermediate 5-35b)

[0696] TIFF2025503429000358.tif23170

[0697] Intermediate 5-35b was synthesized from intermediate 5-35a and tert-butyl(4-aminocyclohexyl)carbamate in the same manner as in Step 2 of Preparation Example 5-40.

[0698] Step 3: methyl(S)-4-((4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]d iazepin-6-yl)acetamido)cyclohexyl)amino)-2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoate (intermediate 5-35'a)

[0699] TIFF2025503429000359.tif46170

[0700] Intermediate 5-35'a was synthesized from Intermediate 5-35b in the same manner as in Steps 4 and 5 of Production Example 4-1'.

[0701] Step 4: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-((4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)cyclohexyl)amino)benzoate trifluoroacetic acid

[0702] Example 5-35' was synthesized from intermediate 5-35'a in the same manner as in Step 5 of Preparation Example 5-85'.

[0703] 1 H NMR (500 MHz, DMSO-d6) δ 8.26 (s, 3H), 8.13 (t, J = 7.2 Hz, 1H), 7.67 (dd, J = 8.8, 4.8 Hz, 1H), 7.53 - 7.32 (m, 4H), 6.77 - 6.53 (m, 2H), 4.46 (dd, J = 8.4, 5.9 Hz, 1H), 3.99 (q, J = 5.7 Hz, 2H), 3.71 (s, 2H), 3.16 - 3.09 (m, 5H) 2.55 (s, 3H), 2.37 (s, 3H), 1.65 (d, J = 23.1 Hz, 8H), 1.98 - 1.84 (m, 2H), 1.57 (d, J = 4.7 Hz, 3H); MS m / z [M+H] + 684.61.

[0704] [Production Example 5-53'](S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoic acid hydrochloride

[0705] TIFF2025503429000360.tif31170

[0706] Step 1: (S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-t rimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoic acid (intermediate 5-53'a)

[0707] TIFF2025503429000361.tif29170

[0708] Intermediate 5-53'a was synthesized from intermediate 4-228'a in the same manner as in Step 1 of Preparation Example 3-2'.

[0709] Step 2: (S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoic acid hydrochloride

[0710] Example 5-53' was synthesized from Intermediate 5-53'a in the same manner as in Step 2 of Preparation Example 1-1.

[0711] 11H NMR (500 MHz, DMSO-d6) δ 10.39 (s, 1H), 8.34 (s, 4H), 8.15 (d, J = 2.3 Hz, 1H), 7.88 (d, J = 8.6 Hz, 1H), 7.46 - 7.41 (m, 5H), 4.51 (dd, J = 8.0, 6.4 Hz, 1H), 4.04 (q, J = 5.7 Hz, 2H), 3.67 (dd, J = 4.8, 1.4 Hz, 2H), 3.26 - 3.23 (m, 2H), 3.21 - 3.17 (m, 2H), 2.60 (s, 3H), 2.41 (s, 3H), 1.78 (d, J = 7.5 Hz, 2H), 1.61 (s, 3H); MS m / z [M+H] + 658.37.

[0712] [Production Example 5-59’] (S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzamide hydrochloride

[0713] TIFF2025503429000362.tif34170

[0714] Step 1: tert-butyl (S)-(3-(2-carbamoyl-5-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)phenyl)prop-2-yn-1-yl)carbamate (Intermediate 5-59’a)

[0715] TIFF2025503429000363.tif31170

[0716] Intermediate 5-53'a (0.04 mmol, 1.0 eq) was dissolved in 1.0 mL of DCM, and a small amount of SOCl2 was added dropwise. The mixture was then stirred at reflux for 1 hour. The reaction temperature was lowered to 0°C, and 2.0 mL of aqueous ammonia was added dropwise. The mixture was then stirred at room temperature for 18 hours. The reaction mixture was dried over Na2SO4 and filtered under reduced pressure. The mixture was evaporated under reduced pressure and purified using a flash column to obtain intermediate 5-59'a.

[0717] Step 2: (S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzamide hydrochloride

[0718] Example 5-59' was synthesized from Intermediate 5-59'a in the same manner as in Step 2 of Preparation Example 1-1.

[0719] 1 H NMR (500 MHz, DMSO-d6) δ 10.27 (s, 1H), 8.31 (dd, J = 11.8, 6.3 Hz, 4H), 8.06 (d, J = 2.2 Hz, 1H), 7.65 (s, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.49 - 7.45 (m, 3H), 7.43 - 7.41 (m, 3H), 4.51 (dd, J = 7.9, 6.4 Hz, 1H), 4.02 (t, J = 5.6 Hz, 2H), 3.50 - 3.48 (m, 2H), 3.21 (ddd, J = 27.8, 11.7, 7.6 Hz, MS m / z [M+H] + 657.44.

[0720] [Production Example 5-61'](S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thien o[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)-N,N-dimethylbenzamide hydrochloride

[0721] TIFF2025503429000364.tif31170

[0722] Step 1: tert-butyl(S)-(3-(5-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a ][1,4]diazepin-6-yl)acetamido)butanamido)-2-(dimethylcarbamoyl)phenyl)prop-2-yn-1-yl)carbamate (intermediate 5-61'a)

[0723] TIFF2025503429000365.tif31170

[0724] Intermediate 5-53'a (0.03 mmol, 1.0 eq) was dissolved in 0.8 mL of dry DCE, and the gas in the reaction vessel was replaced with Ar gas. 0.01 mL of DEA (2.0 M in THF) was added dropwise and stirred at room temperature for 5 minutes. DCC (1.5 eq) and HOBt (1.5 eq) were added and the mixture was reacted at 70°C for 3 hours. The reaction mixture was distilled under reduced pressure and purified using a flash column to synthesize intermediate 5-61'a.

[0725] Step 2: (S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)-N,N-dimethylbenzamide hydrochloride

[0726] Example 5-61' was synthesized from Intermediate 5-61'a in the same manner as in Step 2 of Preparation Example 1-1.

[0727] 1 H NMR (500 MHz, DMSO-d6) δ 10.27 (s, 1H), 8.36 (s, 3H), 8.33 - 8.30 (m, 1H), 8.03 (d, J = 2.1 Hz, 1H), 7.51 (dd, J = 8.4, 2.2 Hz, 1H), 7.46 (d, J = 8.8 Hz, 2H), 7.42 (d, J = 8.6 Hz, 2H), 7.26 (d, J = 8.4 Hz, 1H), 4.54 - 4.51 (m, 1H), 4.01 (d, J = 5.7 Hz, 2H), 3.71 - 3.67 (m, 2H), 3.27 - 3.16 (m, MS m / z [M+H] + 685.61.

[0728] [Production Example 5-63'](S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-methoxyphenyl)-4-(2-(4-(4-chlorophenyl)-2,3, 9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide trifluoroacetic acid

[0729] TIFF2025503429000366.tif31170

[0730] Step 1: tert-butyl(3-(5-amino-2-methoxyphenyl)prop-2-yn-1-yl)carbamate (Intermediate 5-63a)

[0731] TIFF2025503429000367.tif26170

[0732] Intermediate 5-63a was synthesized from 3-iodo-4-methoxyaniline in the same manner as in Step 1 of Production Example 1-1.

[0733] Step 2: tert-butyl(S)-(3-(5-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo [4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)-2-methoxyphenyl)prop-2-yn-1-yl)carbamate (intermediate 5-63'a)

[0734] TIFF2025503429000368.tif31170

[0735] Intermediate 5-63'b was synthesized from intermediates 4-162'a and 5-63a in the same manner as in Step 5 of Preparation Example 4-1'.

[0736] Step 3: (S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-methoxyphenyl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide trifluoroacetic acid

[0737] Example 5-63' was synthesized from Intermediate 5-63'a in the same manner as in Step 2 of Preparation Example 1-1'.

[0738] 1 H NMR (500 MHz, DMSO-d6) δ 9.89 (s, 1H), 8.27 (d, J = 6.5 Hz, 3H), 7.89 (d, J = 2.7 Hz, 1H), 7.47 - 7.39 (m, 6H), 7.04 (d, J = 9.1 Hz, 1H), 4.50 (dd, J = 8.2, 6.2 Hz, 1H), 4.03 (d, J = 5.6 Hz, 2H), 3.78 (s, 3H), 3.20 (ddt, J = 37.0, 23.1, 6.8 Hz, 6H), 2.60 (s, 3H), 2.41 (d, J = 0.9 Hz, 4H), 2.34 (t, J = 7.5 Hz, 2H), 1.79 - 1.73 (m, 2H), 1.62 - 1.61 (m, 3H); MS m / z [M+H] + 644.45.

[0739] [Production Example 5-65'](S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-methylphenyl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide hydrochloride

[0740] TIFF2025503429000369.tif34170

[0741] Example 5-65' was synthesized from 3-iodo-4-methylaniline in the same manner as in Preparation Example 5-77'.

[0742] 1H NMR (500 MHz, DMSO-d6) δ10.162 (s, 1H), 8.531 (brs, 3H), 8.351(t, J = 1.3Hz, 1H), 7.878(d, J = 0.2 Hz), 7.481 - 7.411 (m, 5H), 7.203(d, 1.1 Hz, 1H), 4.572 (t, J = 1.6 Hz, 1H), 4.016 (d, J = 0.2 Hz, 2H), 3.282 - 3.163 (m, 4H), 2.644 (s, 3H), 2.416 (s, 3H), 2.396 - 2.357 (m, 2H), 2.338 (s, 3H), 1.798 - 1.743 (m, 2H), 1.612 (s, 3H); MS m / z [M+H] + 628.4.

[0743] [Production Example 5-75'](S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-(trifluoromethyl)phenyl)-4-(2-(4-(4-chlorophenyl) -2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide hydrochloride

[0744] TIFF2025503429000370.tif36170

[0745] Example 5-75' was synthesized from 3-iodo-4-(trifluoromethyl)aniline in the same manner as in Preparation Example 5-77'.

[0746] 11H NMR (500 MHz, DMSO-d6) δ 10.18 (s, 1H), 9.03 (s, 1H), 8.75 (s, 1H), 8.62 (s, 1H), 8.34 (s, 3H), 8.20 (s, 1H), 7.95 (s, 2H), 7.94 (s, 1H), 7.71 (s, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.25 (d, J = 8.5 Hz, 1H), 7.10 (s, 1H), 7.08 (s, 1H), 4.55 (s, 2H), 3.92 (s, 2H), 3.80 (s, 3H), 3.13 (s, 2H), 2.38 (s, 2H), 1.85 (s, 2H); MS m / z [M+H] + 682.41.

[0747] [Production Example 5-77’] (S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-fluorophenyl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide hydrochloride

[0748] TIFF2025503429000371.tif34170

[0749] Step 1: tert-butyl(3-(5-amino-2-fluorophenyl)prop-2-yn-1-yl)carbamate (Intermediate 5-77a)

[0750] TIFF2025503429000372.tif21170

[0751] 4-fluoro-3-iodoaniline (1.0 mmol, 1.0 eq), n-boc propargylamine (1.5 eq), Pd(PPh3)4 (0.1 eq), and CuI (0.2 eq) were added dropwise to 3.0 mL of TEA. The gas in the reaction vessel was replaced with Ar gas, and the mixture was stirred at 70 °C overnight. The mixture was filtered under reduced pressure through Celite and evaporated. The mixture was purified by flash column chromatography to give intermediate 5-77a.

[0752] Step 2: (S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-fluorophenyl)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide hydrochloride

[0753] Example 5-77' was synthesized from intermediates 4-162'a and 5-77a by applying the same methods as in Step 5 of Preparation Example 4-1' and Step 2 of Preparation Example 1-1 in that order.

[0754] 1 H NMR (400 MHz, DMSO-d6) δ 10.368 (s, 1H), 8.530 (brs, 3H), 8.354 - 8.341 (m, 1H), 7.994 - 7.972 (m, 1H), 7.574 (s, 1H), 7.475 - 7.407 (m, 4H), 7.288 - 7.242 (m, 1H), 4.573 - 4.538 (m, 1H), 4.044 - 4.031 (m, 2H), 3.275 -3.132 (m, 4H), 2.632 (s, 3H), 2.414 (s, 3H), 2.390 - 2.371 (m, 2H), 1.779 - 1.747 (m, 2H), 1.611 (s, 3H); 19 F NMR (400 MHz, DMSO-d6) -117.332; MS m / z [M+H] +632.10.

[0755] [Production Example 5-78']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H -thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)-6-methylbenzoate hydrochloride

[0756] TIFF2025503429000373.tif31170

[0757] Step 1: methyl 4-bromo-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-6-methylbenzoate (Intermediate 5-78a)

[0758] TIFF2025503429000374.tif26170

[0759] Intermediate 5-78a was synthesized from methyl 4-bromo-2-iodo-6-methylbenzoate in the same manner as in Step 1 of Production Example 1-1.

[0760] Step 2: methyl 4-((tert-butoxycarbonyl)amino)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-6-methylbenzoate (intermediate 5-78b)

[0761] TIFF2025503429000375.tif29170

[0762] Intermediate 5-78a (0.5 mmol, 1.0 eq), tert-butyl carbamate (3.0 eq), BrettPhos Pd G3 (0.1 eq), CS2CO3 (3.0 eq), and 1.5 mL of dioxane were mixed and stirred at 100 °C overnight. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column chromatography to give intermediate 5-78b.

[0763] Step 3: methyl 4-amino-2-(3-aminoprop-1-yn-1-yl)-6-methylbenzoate (Intermediate 5-78c)

[0764] TIFF2025503429000376.tif26170

[0765] Intermediate 5-78c was synthesized from intermediate 5-78b in the same manner as in Step 2 of Production Example 1-1.

[0766] Step 4: methyl 4-amino-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-6-methylbenzoate (Intermediate 5-78d)

[0767] TIFF2025503429000377.tif26170

[0768] Intermediate 5-78c (0.1 mmol, 1.0 eq) was dissolved in 3.0 mL of DCM, and then TEA (3.0 eq) and BocO (1.0 eq) were added dropwise at 0 °C and stirred for 2 hours. The reaction mixture was extracted with DCM and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and purified via flash column to give intermediate 5-78d.

[0769] Step 5: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)-6-methylbenzoate hydrochloride

[0770] Example 5-78' was synthesized from intermediate 5-78d in the same manner as in Step 2 of Preparation Example 5-77'.

[0771] 1 H NMR (400 MHz, DMSO-d6) δ 10.38 (s, 1H), 8.50 (s, 3H), 8.34 (t, J = 14 Hz, 1H), 7.86 (s, 1H), 7.48 - 7.41 (m, 5H), 4.56 - 4.53 (m, 1H), 4.00 - 3.98 (m, 2H), 3.86 (s, 3H), 3.27 - 3.15 (m, 2H), 2.62 (s 3H), 2.41 (s, 5H), 2.25 (s, 3H), 1.80 -1.76 (m, 2H), 1.61 (s, 3H); MS m / z [M+H] + 686.4.

[0772] [Production Example 5-81'](S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)-4-(2-(4-(4-chlo rophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide hydrochloride

[0773] TIFF2025503429000378.tif36170

[0774] Step 1: tert-butyl(3-(2-bromo-5-nitrophenyl)prop-2-yn-1-yl)carbamate (5-81a)

[0775] TIFF2025503429000379.tif26170

[0776] Intermediate 5-81a was synthesized from Bromo-2-iodo-4-nitrobenzene in the same manner as in Step 1 of Preparation Example 5-89'.

[0777] Step 2: tert-butyl(3-(5-nitro-2-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)prop-2-yn-1-yl)carbamate (Intermediate 5-81b)

[0778] TIFF2025503429000380.tif29170

[0779] Intermediate 5-81a (3.0 mmol, 1.0 eq), (2-(trifluoromethyl)pyrimidin-5-yl)boronic acid (1.1 eq), Pd(dppf)Cl (0.1 eq), KCO (2.0 eq), and 10.0 mL of dioxane / HO were mixed and stirred at 50 °C for 12 hours. The reaction mixture was purified by flash column chromatography to give intermediate compound 5-81b.

[0780] Step 3: tert-butyl(3-(5-amino-2-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)prop-2-yn-1-yl)carbamate (Intermediate 5-81c)

[0781] TIFF2025503429000381.tif29170

[0782] Intermediate 5-81b (1.5 mmol, 1.0 eq), Fe (3.0 eq), NH4Cl (5.0 eq), and EtOH / H2O were stirred at 80 °C for 2 hours. The reaction mixture was purified by flash column chromatography to give Intermediate 5-81c.

[0783] Step 4: (S)-N-(3-(3-aminoprop-1-yn-1-yl)-4-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)-4-(2-(4-(4-chloro phenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamide hydrochloride

[0784] Example 5-81' was synthesized from Intermediate 5-81c in the same manner as in Steps 2 and 3 of Preparation Example 5-77'.

[0785] 1 H NMR (500 MHz, DMSO-d6) δ 10.36 (s, 1H), 9.25 (s, 2H), 8.28 (dd, J = 13.5, 7.6 Hz, 4H), 8.18 (t, J = 1.3 Hz, 1H), 7.62 (d, J = 1.8 Hz, 2H), 7.42 (d, J = 8.9 Hz, 2H), 7.40 - 7.35 (m, 2H), 4.48 (dd, J = 8.1, 6.2 Hz, 1H), 3.91 (q, J = 5.6 Hz, 2H), 3.51 (s, 1H), 3.27 - 3.07 (m, 4H), 2.56 (s, 3H), 2.43 - 2.34 (m, 5H), 1.76 (q, J = 7.3 Hz, 2H), 1.58 (d, J = 0.9 Hz, 3H); MS m / z [M+H] + 760.44.

[0786] [Production Example 5-89']methyl(S)-3-(3-aminoprop-1-yn-1-yl)-5-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoate hydrochloride

[0787] TIFF2025503429000382.tif36170

[0788] Step 1: methyl 3-amino-5-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate (Intermediate 5-89a)

[0789] TIFF2025503429000383.tif33170

[0790] Methyl 3-amino-5-bromobenzoate (4.0 mmol, 1.0 eq), n-boc propargylamine (2.0 eq), Xphos Pd G4 (0.03 eq), CuI (0.1 eq), Cs2CO3 (3.0 eq), and 10.0 mL of dry toluene were mixed and stirred at 100 °C for 12 hours. The reaction mixture was filtered through Celite under reduced pressure. The mixture was evaporated under reduced pressure and purified by flash column chromatography to give intermediate 5-89a.

[0791] Step 2: methyl(S)-3-(3-aminoprop-1-yn-1-yl)-5-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoate hydrochloride

[0792] Example 5-89' was synthesized from Intermediate 5-89a in the same manner as in Steps 2 and 3 of Preparation Example 5-77'.

[0793] 1 H NMR (500 MHz, DMSO-d6) δ 10.26 (s, 1H), 8.26 (t, J= 5.8 Hz, 1H), 8.13 (s, 1H), 7.96 (s, 1H), 7.54 (s, 1H), 7.44 - 7.35 (m, 6H), 4.46 (dd, J= 8.3, 6.0 Hz, 1H), 3.81 (s, 3H), 3.57 (s, 3H), 3.17 (d, J= 6.4 Hz, 2H), 3.11 - 3.08 (m, 2H), 2.55 (s, 3H), 2.36 (s, 3H), 1.74 (q, J= 7.2 Hz, 3H), 1.57 (s, 4H); MS m / z [M+H] + 672.52.

[0794] [Production Example 5-90']methyl(S)-4-(3-aminoprop-1-yn-1-yl)-2-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoate hydrochloride

[0795] TIFF2025503429000384.tif39170

[0796] Example 5-90' was synthesized from methyl 2-amino-4-iodobenzoate in the same manner as in Example 5-77'.

[0797] 11H NMR (500 MHz, DMSO-d6) δ 10.61 (s, 1H), 8.36 (d, J = 1.7 Hz, 1H), 8.29 (t, J = 5.7 Hz, 1H), 7.87 (d, J = 8.2 Hz, 1H), 7.49 - 7.38 (m, 6H), 7.16 (dd, J = 8.2, 1.7 Hz, 1H), 4.49 (dd, J = 8.3, 5.9 Hz, 1H), 3.81 (s, 3H), 3.52 (s, 2H), 3.26 - 3.22 (m, 2H), 3.14 (ddd, J = 16.9, 7.9, 5.6 Hz, 2H), 2.58 (d, J = 2.6 Hz, 3H), 2.40 (d, J = 0.9 Hz, 4H), 1.80 (dd, J = 13.9, 6.7 Hz, 3H), 1.61 (t, J = 1.2 Hz, 4H); MS m / z [M+H] + 672.52.

[0798] [Production Example 5-96'] methyl (S)-3-(3-aminoprop-1-yn-1-yl)-6-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)picolinate hydrochloride

[0799] TIFF2025503429000385.tif34170

[0800] Step 1: methyl 6-amino-3-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)picolinate (Intermediate 5-96a)

[0801] TIFF2025503429000386.tif18170

[0802] Intermediate 5-96a was synthesized in the same manner as in Step 1 of Preparation Example 5-89′ using methyl 6-amino-3-bromopicolinate.

[0803] Step 2: methyl(S)-3-(3-aminoprop-1-yn-1-yl)-6-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)picolinate hydrochloride

[0804] Example 5-96' was synthesized from Intermediate 5-96a in the same manner as in Steps 2 and 3 of Preparation Example 5-77'.

[0805] 1 H NMR (500 MHz, DMSO-d6) δ 10.99 (s, 1H), 8.30 (d, J = 8.8 Hz, 1H), 8.26 (s, 4H), 8.22 (t, J = 5.7 Hz, 1H), 7.97 (d, J = 8.7 Hz, 1H), 7.45 - 7.39 (m, 2H), 7.39 - 7.33 (m, 2H), 4.46 (dd, J = 8.3, 6.0 Hz, 1H), 4.02 (q, J = 5.6 Hz, 2H), 3.85 (s, 3H), 3.26 - 3.04 (m, 4H), 2.55 (s, 5H), 2.37 (d, J = 0.8 MS m / z [M+H] + 673.42.

[0806] [Production Example 5-104'](S)-N-(3-(3-aminoprop-1-yn-1-yl)phenyl)-6-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)hexanamide hydrochloride

[0807] TIFF2025503429000387.tif48170

[0808] Example 5-104' was synthesized from Intermediate 4-165'a and Intermediate 1-3a in the same manner as in Step 2 of Preparation Example 5-77'.

[0809] 1 H NMR (500 MHz, DMSO-d6) δ 10.09 (s, 1H), 8.35 (s, 3H), 8.19 (s, 1H), 7.92 (s, 1H), 7.46 (d, J = 8.8 Hz, 4H), 7.38 (d, J = 8.5 Hz, 2H), 7.29 (t, J = 7.9 Hz, 1H), 7.07 (d, J = 10.1 Hz, 1H), 4.50 - 4.46 (m, 1H), 3.96 (t, J = 5.6 Hz, 2H), 3.24 - 3.12 (m, 2H), 3.10 - 3.03 (m, 2H), 2.57 (s, 3H), 2.37 (s, 3H), 2.28 (t, J = 7.5 Hz, 2H), 1.57 (d, J = 7.2 Hz, 6H), 1.47 - 1.40 (m, 2H), 1.34 - 1.27 (m, 2H); MS m / z [M+H] + 642.45.

[0810] [Production Example 5-106']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl -6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethoxy)-5-oxopentanamido)benzoate hydrochloride

[0811] TIFF2025503429000388.tif45170

[0812] Step 1: methyl(S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetate (Intermediate 5-106a)

[0813] TIFF2025503429000389.tif31170

[0814] JQ1 carboxylic acid (1.5 mmol, 1.0 eq) was dissolved in 8.0 mL of MeOH, and then SOCl2 (3.0 eq) was added dropwise. After stirring at room temperature for 3 hours, the solvent was removed under reduced pressure, followed by the dropwise addition of DCM, and the remaining solvent was removed again under reduced pressure. The compound was dissolved in a small amount of EtOAc, and the resulting solid was recrystallized using n-hexane. The resulting solid was filtered, and intermediate 5-106a was synthesized without further purification.

[0815] Step 2: (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethan-1-ol (Intermediate 5-106'b)

[0816] TIFF2025503429000390.tif31170

[0817] Intermediate 5-106'a (0.3 mmol, 1.0 eq) was dissolved in 3.0 mL of dry THF and stirred at 0 °C. 1.0 M lithium aluminum hydride solution in THF (1.5 eq) was slowly added dropwise over 10 min. After 5 min, ice water was added to terminate the reaction. 2 N aqueous HCl was added dropwise to the mixture to acidify it to pH 3. After extraction with EtOAc and H2O, the organic layer was washed with NaHCO3 and aqueous NaCl. The organic layer was dried over Na2SO4, filtered under reduced pressure, and evaporated to give intermediate 5-106'b without further purification.

[0818] Step 3: (S)-5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethoxy)-5-oxopentanoic acid (Intermediate 5-106'c)

[0819] TIFF2025503429000391.tif34170

[0820] Intermediate 5-106'b (0.1 mmol, 1.0 eq) and glutaric acid (2.0 eq) were dissolved in dry DCM and stirred at room temperature. DCC (1.1 eq) and DMAP (1.1 eq) were added dropwise and the mixture was stirred for 18 hours. The precipitate formed after the reaction was filtered, and the filtrate was purified by flash column to synthesize intermediate 5-106'c.

[0821] Step 4: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(5-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethoxy)-5-oxopentanamido)benzoate hydrochloride

[0822] Example 5-106' was synthesized from Intermediate 5-106'c and Intermediate 3-25a in the same manner as in Step 2 of Preparation Example 5-77'.

[0823] 1 H NMR (500 MHz, DMSO-d6) δ 7.89 - 7.82 (m, 1H), 7.50 - 7.45 (m, 7H), 7.21 (s, 1H), 6.67 (s, 1H), 4.44 - 4.39 (m, 2H), 4.18 - 4.15 (m, 1H), 4.05 (s, 2H), 3.79 (s, 3H), 2.65 - 2.63 (m, 2H), 2.59 (s, 3H), 2.36 (s, 3H), 2.18 (t, J = 7.3 Hz, 2H), 2.02 (s, 2H), 1.60 (s, 3H); MS m / z [M+H] + 687.31.

[0824] [Production Example 5-107'](S)-N-(3-(3-aminoprop-1-yn-1-yl)phenyl)-8-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)octanamide hydrochloride

[0825] TIFF2025503429000392.tif48170

[0826] Step 1: methyl(S)-8-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)octanoate (Intermediate 5-107a)

[0827] TIFF2025503429000393.tif31170

[0828] Intermediate 5-107'a was synthesized from methyl 8-aminooctanoate in the same manner as in Step 5 of Preparation Example 4-1'.

[0829] Step 2: (S)-8-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)octanoic acid (Intermediate 5-107b)

[0830] TIFF2025503429000394.tif34170

[0831] Intermediate 5-107'b was synthesized from intermediate 5-107'a in the same manner as in step 1 of 3-2'.

[0832] Step 3: (S)-N-(3-(3-aminoprop-1-yn-1-yl)phenyl)-8-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)octanamide hydrochloride

[0833] Example 5-107' was synthesized from Intermediate 5-107'b and Intermediate 1-3a in the same manner as in Step 2 of Preparation Example 5-77'.

[0834] 1H NMR (500 MHz, DMSO-d6) δ 10.12 (s, 1H), 8.42 (s, 3H), 8.17 (t, J = 5.6 Hz, 1H), 7.90 (t, J = 1.9 Hz, 1H), 7.50 - 7.43 (m, 3H), 7.41 - 7.36 (m, 2H), 7.28 (t, J = 7.9 Hz, 1H), 7.07 (dt, J = 7.7, 1.3 Hz, 1H), 4.49 (dd, J = 8.0, 6.1 Hz, 1H), 3.95 (q, J = 5.7 Hz, 2H), 3.24 - 3.12 (m, 2H), 3.06 (dq, J = 22.0, 6.4 Hz, 2H), 2.58 (s, 3H), 2.37 (d, J = 1.0 Hz, 3H), 2.27 (t, J = 7.5 Hz, 2H), 1.58 (d, J = 1.0 Hz, 3H), 1.56 - 1.50 (m, 2H), 1.42 - 1.36 (m, 2H), 1.25 (s, 6H); MS m / z [M+H] + 670.53.

[0835] [Production Example 5-108'](S)-N-(3-(3-aminoprop-1-yn-1-yl)phenyl)-11-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)undecanamide hydrochloride

[0836] TIFF2025503429000395.tif53170

[0837] Methyl 11-aminoundecanoate is produced by the same method as in Production Example 5-107, and is synthesized in Example 5-108.

[0838] 11H NMR (500 MHz, DMSO-d6) δ 10.05 (s, 1H), 8.34 (s, 3H), 8.15 (t, J = 5.6 Hz, 1H), 7.91 (s, 1H), 7.44 (d, J = 8.8 Hz, 4H), 7.38 (d, J = 8.6 Hz, 2H), 7.29 (t, J = 7.9 Hz, 1H), 7.07 (d, J = 7.7 Hz, 1H), 4.50 - 4.45 (m, 1H), 3.96 (q, J = 5.7 Hz, 2H), 3.25 - 3.17 (m, 2H), 3.15 - 3.05 (m, 2H), 2.56 (s, 3H), 2.37 (s, 3H), 2.26 (t, J = 7.5 Hz, 2H), 1.58 (s, 3H), 1.56 - 1.50 (m, 2H), 1.41 - 1.36 (m, 2H), 1.22 (s, 14H); MS m / z [M+H] + 712.58.

[0839] [Production Example 5-109’] (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethyl 11-((3-(3-aminoprop-1-yn-1-yl)phenyl)amino)-11-oxoundecanoate hydrochloride

[0840] TIFF2025503429000396.tif50170

[0841] Using intermediate 5-106'b, undecanedioic acid, and intermediate 1-3a, Example 5-109' was synthesized via intermediate 5-109'a ((S)-11-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethoxy)-11-oxoundecanoic acid) in the same manner as in steps 3 and 4 of Preparation Example 5-106'.

[0842] 1 H NMR (500 MHz, DMSO-d6) δ 10.03 (s, 1H), 8.28 (s, 3H), 7.97 (d, J = 2.0 Hz, 1H), 7.49 (t, J = 9.1 Hz, 6H), 7.33 (t, J = 7.9 Hz, 1H), 7.11 (dt, J = 7.6, 1.3 Hz, 1H), 4.48 - 4.42 (m, 1H), 4.38 (dt, J = 11.1, 6.2 Hz, 1H), 4.14 (dd, J = 8.6, 5.6 Hz, 1H), 4.01 (q, J = 5.6 Hz, 3H), 2.59 (s, 3H), 2.39 (d, J = 0.9 Hz, 4H), 2.29 (d, J = 7.3 Hz, 2H), 2.26 (d, J = 7.2 Hz, 2H), 1.61 (d, J = 0.9 Hz, 3H), 1.56 (s, 2H), 1.44 (s, 2H), 1.26 (d, J = 7.8 Hz, 4H), 1.18 (s, 6H); MS m / z [M+H] + 713.58.

[0843] [Production Example 5-110']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(11-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl- 6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethoxy)-11-oxoundecanamido)benzoate hydrochloride

[0844] TIFF2025503429000397.tif43170

[0845] Intermediate 5-109'a and Intermediate 3-25a were synthesized using the same method as in Production Example 5-77' and Example 5-110' was used.

[0846] 1 H NMR (500 MHz, DMSO-d6) δ 10.37 (d, J = 16.6 Hz, 1H), 8.35 (s, 2H), 8.09 (q, J = 2.4 Hz, 1H), 7.88 - 7.83 (m, 1H), 7.54 - 7.38 (m, 5H), 4.41 (dt, J = 10.8, 7.0 Hz, 1H), 4.34 (dt, J = 11.2, 6.1 Hz, 1H), 4.11 (dd, J = 8.6, 5.6 Hz, 1H), 4.04 - 3.98 (m, 2H), 3.79 (s, 3H), 2.64 - 2.58 (m, 2H), 2.56 (d, J = 1.3 Hz, 3H), 2.35 (s, 3H), 2.30 (t, J = 7.3 Hz, 2H), 2.21 (t, J = 7.3 Hz, 2H), 1.57 (s, 3H), 1.52 (t, J = 7.2 Hz, 2H), 1.39 (d, J = 8.0 Hz, 2H), 1.20 (s, 4H), 1.14 (s, 6H); MS m / z [M+H] + 771.53.

[0847] [Production Example 5-115']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetoxy)butanamido)benzoate hydrochloride

[0848] TIFF2025503429000398.tif37170

[0849] Step 1: tert-butyl(S)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetoxy)butanoate (Intermediate 5-115a)

[0850] TIFF2025503429000399.tif29170

[0851] Intermediate 5-115'a was synthesized from tert-butyl 4-hydroxybutanoate in the same manner as in Step 5 of Preparation Example 4-1'.

[0852] Step 2: (S)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetoxy)butanoic acid (Intermediate 5-115b)

[0853] TIFF2025503429000400.tif34170

[0854] Intermediate 5-115'b was synthesized from intermediate 5-115'a in the same manner as in Step 4 of Preparation Example 4-1'.

[0855] Step 3: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetoxy)butanamido)benzoate hydrochloride

[0856] Example 5-115' was synthesized from Intermediate 5-115'b and Intermediate 3-25a in the same manner as in Steps 2 and 3 of Preparation Example 5-77'.

[0857] 1 H NMR (500 MHz, DMSO-d6) δ 10.40 (s, 1H), 8.31 (s, 2H), 8.15 (d, J = 2.1 Hz, 1H), 7.89 (d, J = 8.7 Hz, 1H), 7.53 - 7.39 (m, 7H), 4.51 - 4.44 (m, 2H), 4.16 (d, J = 5.9 Hz, 3H), 4.07 (d, J = 5.6 Hz, 3H), 3.83 (d, J = 1.5 Hz, 3H), 3.65 - 3.59 (m, 1H), 3.47 - 3.37 (m, 3H), 3.14 (dd, J = 7.3, 4.3 Hz, MS m / z [M+H] + 673.42.

[0858] [Production Example 5-148']methyl(S)-2-(3-acetamidoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)butanamido)benzoate

[0859] TIFF2025503429000401.tif33170

[0860] Example 4-228' (0.02 mmol, 1.0 eq) was dissolved in 0.6 mL of DCM, and then 9 μL of TEA and 2.4 μL of AcO were added dropwise at 0°C and stirred at room temperature for 40 minutes. The reactant was distilled under reduced pressure and purified by flash column to synthesize Example 5-148'.

[0861] 1 H NMR (400 MHz, DMSO-d6) δ 10.31 (s, 1H), 8.42 (t, J = 5.5 Hz, 1H), 8.30 (t, J = 5.7 Hz, 1H), 7.91 (d, J = 2.2 Hz, 1H), 7.84 (d, J = 8.7 Hz, 1H), 7.60 (dd, J = 8.7, 2.2 Hz, 1H), 7.47 - 7.43 (m, 2H), 7.43 - 7.39 (m, 2H), 4.51 (dd, J = 8.1, 6.2 Hz, 1H), 4.13 (d, J = 5.5 Hz, 2H), 3.80 (s, 3H), 3.28 - 3.13 (m, 4H), 2.60 (s, 3H), 2.43 - 2.39 (m, 5H), 1.86 (s, 3H), 1.77 (p, J = 7.6 Hz, 2H), 1.61 (d, J = 0.9 Hz, 3H); MS m / z [M+H] + 714.48.

[0862] [Production Example 5-149']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H -thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-N-methylacetamido)butanamido)benzoate hydrochloride

[0863] TIFF2025503429000402.tif48170

[0864] Step 1: (S)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-N-methylacetamido)butanoic acid (Intermediate 5-149a)

[0865] TIFF2025503429000403.tif34170

[0866] Intermediate 5-149'a was synthesized from methyl 4-(methylamino)butanoate hydrochloride by the same methods as in Step 5 of Production Example 4-1' and Step 1 of Production Example 3-2'.

[0867] Step 2: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-N-methylacetamido)butanamido)benzoate hydrochloride

[0868] Example 5-149' was synthesized from Intermediate 5-149'a and Intermediate 3-25a in the same manner as in Step 2 of Preparation Example 5-77'.

[0869] 1 H NMR (400 MHz, DMSO-d6) δ 10.667 - 10.521 (m, 1H), 8.566 (brs, 3H), 8.160 - 8.138 (m, 1H), 7.889 - 7.823 (m, 1H), 7.525 - 7.512 (m, 1H). 7.508 - 7.409 (m, 4H), 4.572 (m, 1H), 4.052 - 4.032 (m, 2H), 3.841 (s, 3H), 3.713 - 3.501 (m, 2H), 3.407 - 3.391 (m, 2H), 3.148 (s, 3H), 2.640 (s, 3H), 2.499 (s, 3H), 2.430 - 2.360 (m, 2H), 1.827 - 1.810 (m, 1H), 1.793 - 1.635 (m, 1H), 1.626 (s, 3H); MS m / z [M+H] + 686.2.

[0870] [Production Example 5-150'] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)-N-methylbutanamido)benzoate hydrochloride

[0871] TIFF2025503429000404.tif42170

[0872] Step 1: methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(methylamino)benzoate (Intermediate 5-150a)

[0873] TIFF2025503429000405.tif26170

[0874] Intermediate 3-25a (0.6 mmol, 1.0 eq), MeB(OH)2 (2.5 eq), Cu(OAc)2 (2.5 eq), pyridine (3.5 eq), and 6.0 mL of dioxane were mixed, and the gas in the reaction vessel was replaced with Ar gas. The reaction mixture was stirred at 110 °C overnight. The reaction mixture was filtered under reduced pressure through celite and then evaporated. The mixture was purified by flash column chromatography to synthesize intermediate 5-150a.

[0875] Step 2: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)-N-methylbutanamido)benzoate hydrochloride

[0876] Example 5-150' was synthesized from intermediate 5-150a in the same manner as in Step 2 of Example 5-77'.

[0877] 1 H NMR (400 MHz, DMSO-d6) δ 8.57 (brs, 3H), 8.26 (t, J= 13 Hz, 1H), 7.93 (d, J = 21 Hz, 1H), 7.61 (m, 1H), 7.49 (m, 3H), 7.42 (m, 2H), 4.54 (m, 1H), 4.02 (m, 2H), 3.88 (s, 3H), 3.25 (m, 5H), 3.16 (m, 2H), 2.64 (s, 3H), 2.42 (s, 3H), 2.33 (m, 2H), 1.68 (m, 2H), 1.62 (s, 3H); MS m / z [M+H] + 686.3.

[0878] [Manufacturing example III'] A TBM-UBM compound in which a compound (JQ1) exhibiting activity as a BRD4 (bromodomain-containing protein 4) inhibitor is linked to a compound corresponding to formula III of the present invention was synthesized as follows.

[0879] [Production Example 4-2']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-5-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl -6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazin-1-yl)benzoate hydrochloride

[0880] TIFF2025503429000406.tif29170

[0881] Step 1: tert-butyl 4-(3-(methoxycarbonyl)phenyl)piperazine-1-carboxylate (Intermediate 4-2a)

[0882] TIFF2025503429000407.tif16170

[0883] Intermediate 4-2a was synthesized from methyl 3-iodobenzoate in the same manner as in Step 1 of Preparation Example 4-226'.

[0884] Step 2: tert-butyl 4-(4-bromo-3-(methoxycarbonyl)phenyl)piperazine-1-carboxylate (Intermediate 4-2b)

[0885] TIFF2025503429000408.tif18170

[0886] Intermediate 4-2a (2.0 mmol, 1.0 eq) was dissolved in dry DCM, and then NBS (1.2 eq) was added and stirred at room temperature for 30 minutes. The reaction mixture was extracted with DCM and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and purified via flash column to give intermediate 4-2b.

[0887] Step 3: methyl 2-bromo-5-(piperazin-1-yl)benzoate hydrochloride (Intermediate 4-2c)

[0888] TIFF2025503429000409.tif16170

[0889] Intermediate 4-2c was synthesized from intermediate 4-2b in the same manner as in Step 2 of Production Example 1-1.

[0890] Step 4: methyl(S)-2-bromo-5-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazin-1-yl)benzoate (intermediate 4-2'a)

[0891] TIFF2025503429000410.tif34170

[0892] Intermediate 4-2'a was synthesized from intermediate 4-2c in the same manner as in Step 5 of Production Example 4-1'.

[0893] Step 5: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-5-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazin-1-yl)benzoate hydrochloride

[0894] Example 4-2' was synthesized from Intermediate 4-2'a in the same manner as in Steps 4 and 5 of Production Example 5-140'.

[0895] MS m / z [M+H] + 656.41.

[0896] [Production Example 4-226'](S)-1-(4-(4-(3-aminoprop-1-yn-1-yl)phenyl)piperazin-1-yl)-2-(4-(4-chlorophen yl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethane-1-one hydrochloride

[0897] TIFF2025503429000411.tif31170

[0898] Step 1: tert-butyl 4-phenylpiperazine-1-carboxylate (Intermediate 4-226a)

[0899] TIFF2025503429000412.tif18170

[0900] Iodobenzene (1.4 mmol, 1.0 eq), tert-butyl piperazine-1-carboxylate (1.1 eq), Pd(OH) (0.1 eq), BINAP (0.1 eq), CSCO (2.0 eq), and dry toluene were mixed, and the gas in the reaction vessel was replaced with Ar gas. The mixture was then stirred overnight at 100°C. The reaction mixture was filtered through Celite, concentrated under reduced pressure, and purified using a flash column to synthesize intermediate 4-226a.

[0901] Step 2: tert-butyl 4-(4-iodophenyl)piperazine-1-carboxylate (Intermediate 4-226b)

[0902] TIFF2025503429000413.tif18170

[0903] Intermediate 4-226a (2.0 mmol, 1.0 eq) was dissolved in dry DCM, and then NIS (1.2 eq) was added and stirred at room temperature for 30 minutes. The reaction mixture was extracted with DCM and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and purified via flash column to give intermediate 4-226b.

[0904] Step 3: 1-(4-iodophenyl)piperazine hydrochloride (Intermediate 4-226c)

[0905] TIFF2025503429000414.tif18170

[0906] Intermediate 4-226c was synthesized from Intermediate 4-226b in the same manner as in Step 2 of Production Example 1-1.

[0907] Step 4: (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-1-(4-(4-iodophenyl)piperazin-1-yl)ethan-1-one (Intermediate 4-226a)

[0908] TIFF2025503429000415.tif36170

[0909] Intermediate 4-226'a was synthesized from intermediate 4-226c in the same manner as in Step 5 of Preparation Example 4-1'.

[0910] Step 5: (S)-1-(4-(4-(3-aminoprop-1-yn-1-yl)phenyl)piperazin-1-yl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethan-1-one hydrochloride

[0911] Example 4-226' was synthesized from Intermediate 4-226'a in the same manner as in Preparation Example 1-1.

[0912] 1 H NMR (500 MHz, DMSO-d6) δ 8.34 - 8.12 (s, 3H), 7.48 - 7.39 (m, 4H), 7.33 - 7.25 (m, 1H), 7.03 - 6.98 (m, 1H), 6.97 - 6.94 (m, 1H), 4.59 (tt, J = 6.9, 2.6 Hz, 1H), 4.06 - 4.02 (m, 2H), 3.82 - 3.76 (m, 1H), 3.66 - 3.56 (m, 2H), 3.53 (s, 3H), 3.51 - 3.41 (m, 2H), 3.40 - 3.28 (m, 2H), 3.19 (d, J = 26.3 Hz, 2H), 2.60 (s, 3H), 2.38 (s, 3H), 1.59 (s, 3H); MS m / z [M+H] + 598.45.

[0913] [Production Example 4-227'](S)-1-(4-(5-(3-aminoprop-1-yn-1-yl)furan-2-carbonyl)piperazin-1-yl)-2-(4-(4-chlor diazepin-6-yl)ethane-1-one hydrochloride

[0914] TIFF2025503429000416.tif34170

[0915] Step 1: methyl 5-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)furan-2-carboxylate (Intermediate 4-227a)

[0916] TIFF2025503429000417.tif13170

[0917] Intermediate 4-227a was synthesized from methyl 5-bromo-2-furancarboxylate in the same manner as in Step 2 of Preparation Example 4-179′.

[0918] Step 2: 5-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)furan-2-carboxylic acid (Intermediate 4-227b)

[0919] TIFF2025503429000418.tif13170

[0920] Intermediate 4-227b was synthesized from intermediate 4-227a in the same manner as in Step 1 of Production Example 3-2'.

[0921] Step 3: (S)-1-(4-(5-(3-aminoprop-1-yn-1-yl)furan-2-carbonyl)piperazin-1-yl)-2-(4-(4-chloroph enyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ethan-1-one hydrochloride

[0922] Example 4-227' was synthesized from Intermediate 5-140'b and Intermediate 4-227b in the same manner as in Preparation Example 5-7'.

[0923] 11H NMR (500 MHz, DMSO-d6) δ 8.41 (s, 3H), 7.51 - 7.45 (m, 2H), 7.45 - 7.38 (m, 2H), 7.09 (d, J = 3.6 Hz, 1H), 7.01 (d, J = 3.6 Hz, 1H), 4.56 (t, J = 6.7 Hz, 1H), 4.06 (q, J = 5.7 Hz, 2H), 3.75 (d, J = 4.4 Hz, 2H), 3.66 - 3.62 (m, 4H), 3.43 - 3.36 (m, 4H), 2.57 (s, 3H), 2.38 (s, 3H), 1.59 (s, 3H); MS m / z [M+H] + 616.33.

[0924] [Production Example 5-7'] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazin-1-yl)benzoate hydrochloride

[0925] TIFF2025503429000419.tif31170

[0926] Step 1: methyl (S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazin-1-yl)benzoate (Intermediate 5-7'a)

[0927] TIFF2025503429000420.tif34170

[0928] Intermediate 5-7'a was synthesized from intermediate 5-7b in the same manner as in Step 5 of Production Example 4-1'.

[0929] Step 2: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazin-1-yl)benzoate hydrochloride

[0930] Example 5-7' was synthesized from Intermediate 5-7'a in the same manner as in Step 2 of Preparation Example 1-1.

[0931] 1 H NMR (500 MHz, DMSO-d6) δ 8.33 (s, 3H), 7.84 (d, J = 9.0 Hz, 1H), 7.48 (d, J = 8.8 Hz, 2H), 7.43 (d, J = 8.7 Hz, 2H), 7.10 - 7.07 (m, 1H), 7.03 (d, J = 2.6 Hz, 1H), 4.60 (t, J = 6.8 Hz, 1H), 4.04 (d, J = 5.7 Hz, 2H), 3.80 (s, 3H), 3.71 - 3.61 (m, 8H), 3.39 - 3.34 (m, 2H), 2.60 (d, J = 1.1 Hz, 3H), 2.42 (d, J = 0.9 Hz, 3H), 1.63 (d, J = 1.0 Hz, 3H); MS m / z [M+H] + 656.42.

[0932] [Production Example 5-22']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H- thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)carbamoyl)benzoate hydrochloride

[0933] TIFF2025503429000421.tif30170

[0934] Step 1: tert-butyl 4-(3-bromo-4-(methoxycarbonyl)benzamido)piperidine-1-carboxylate (Intermediate 5-22a)

[0935] TIFF2025503429000422.tif15170

[0936] Intermediate 5-22a was synthesized in the same manner as in Step 5 of Production Example 4-1′ using 3-Bromo-4-(methoxycarbonyl)benzoic acid (0.2 mmol, 1.0 eq) and tert-butyl 4-aminopiperidine-1-carboxylate as starting materials.

[0937] Step 2: methyl 2-bromo-4-(piperidin-4-ylcarbamoyl)benzoate hydrochloride (Intermediate 5-22b)

[0938] TIFF2025503429000423.tif15170

[0939] Intermediate 5-22b was synthesized from Intermediate 5-22a in the same manner as in Step 2 of Production Example 1-1.

[0940] Step 3: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-th ieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)carbamoyl)benzoate hydrochloride

[0941] Intermediate 5-22'a (methyl(S)-2-bromo-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)carbamoyl)benzoate) was synthesized from intermediate 5-22b in the same manner as in steps 4 and 5 of Preparation Example 4-2', and then Example 5-22' was synthesized.

[0942] 1H NMR (500 MHz, DMSO-d6) δ 8.75 (dd, J = 7.6, 3.6 Hz, 1H), 8.56 (s, 3H), 8.10 (dd, J = 5.2, 1.7 Hz, 1H), 8.04 - 7.96 (m, 2H), 7.53 - 7.50 (m, 2H), 7.46 (dd, J = 8.6, 4.8 Hz, 2H), 4.65 - 4.62 (m, 1H), 4.35 (d, J = 13.0 Hz, 1H), 4.17 (s, 1H), 4.06 (d, J = 5.7 Hz, 2H), 3.91 (d, J = 1.5 Hz, 3H), 3.68 (dd, J = 16.5, 7.5 Hz, 1H), 3.44 (dt, J = 16.4, 6.6 Hz, 1H), 3.28 (t, J = 12.6 Hz, 1H), 2.79 (q, J = 12.4, 12.0 Hz, 1H), 2.64 (s, 3H), MS m / z [M+H] + 698.50.

[0943] [Production Example 5-23']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(1-(2-(4-(4-chlorophenyl))-2,3,9-trimethyl-6H- thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)benzoate hydrochloride

[0944] TIFF2025503429000424.tif36170

[0945] Intermediate 5-25'b and Intermediate 3-25a were synthesized by the same method as Production Example 5-7' and Example 5-23'.

[0946] 1 1H NMR (500 MHz, DMSO-d6) δ 10.57 (d, J = 8.6 Hz, 1H), 8.41 (s, 3H), 8.17 (dd, J = 20.9, 2.2 Hz, 1H), 7.91 (dd, J = 8.7, 3.5 Hz, 1H), 7.61 (ddd, J = 24.0, 8.7, 2.2 Hz, 1H), 7.51 (d, J = 8.7 Hz, 2H), 7.45 (t, J = 8.9 Hz, 2H), 4.60 (t, J = 6.7 Hz, 1H), 4.42 (d, J = 3.7 Hz, 1H), 4.23 (d, J = 13.5 Hz, 1H), 4.06 (q, J = 5.6 Hz, 2H), 3.84 (d, J = 1.8 Hz, 3H), 3.64 (ddd, J = 20.5, 16.4, 7.4 Hz, 1H), 3.50 - 3.34 (m, 1H), 3.22 (q, J = 11.5 Hz, 1H), 2.71 (t, J = 12.5 Hz, 2H), 2.61 (s, 3H), 2.42 (s, �H), 1.95 (d, J = 13.3 Hz, 1H), 1.84 (s, 1H), 1.70 (t, J = 11.0 Hz, 1H), 1.64 (s, 3H), 1.52 - 1.42 (m, 1H); MS m / z [M+H] + 698.3.

[0947] [Production Example 5-25’] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-(3-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)propanamido)benzoate hydrochloride

[0948] TIFF2025503429000425.tif30170

[0949] Step 1: 3-(1,3-dioxoisoindolin-2-yl)propanoic acid (Intermediate 5-25a)

[0950] TIFF2025503429000426.tif18170

[0951] β-Alanine (2.0 mmol, 2.0 eq) and phthalic anhydride (1.0 eq) were dissolved in 0.6 mL of acetic acid and stirred at 120 °C overnight. The reaction mixture was extracted with DCM and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and purified via flash column to synthesize intermediate 5-25a.

[0952] Step 2: methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(3-(1,3-dioxoisoindolin-2-yl)propanamido)benzoate (Intermediate 5-25b)

[0953] TIFF2025503429000427.tif21170

[0954] Intermediate 5-25b was synthesized from Intermediate 5-25a and Intermediate 3-25a in the same manner as in Step 5 of Production Example 4-1'.

[0955] Step 3: methyl 4-(3-aminopropanamido)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate (Intermediate 5-25c)

[0956] TIFF2025503429000428.tif18170

[0957] Intermediate 5-25c was synthesized from intermediate 5-25b in the same manner as in Step 3 of Preparation Example 5-116.

[0958] Step 4: methyl(S)-1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxylate (Intermediate 5-25a)

[0959] TIFF2025503429000429.tif34170

[0960] Intermediate 5-25'a was synthesized from methyl piperidine-4-carboxylate in the same manner as in Step 5 of Preparation Example 4-1'.

[0961] Step 5: (S)-1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxylic acid (Intermediate 5-25'b)

[0962] TIFF2025503429000430.tif31170

[0963] Intermediate 5-25'b was synthesized from intermediate 5-25'a in the same manner as in Step 1 of Production Example 3-2'.

[0964] Step 6: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(3-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[ 3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)propanamido)benzoate hydrochloride

[0965] Intermediate 5-25'b and Intermediate 5-25c were synthesized by the same method as Production Example 5-7' and Example 5-25'.

[0966] 1 H NMR (500 MHz, DMSO-d6) δ 10.44 (s, 1H), 8.34 (s, 3H), 8.13 (t, J = 2.3 Hz, 1H), 8.00 (dd, J = 10.2, 5.4 Hz, 1H), 7.90 (d, J = 8.7 Hz, 1H), 7.56 (dd, J = 8.7, 2.2 Hz, 1H), 7.50 (d, J = 4.7 Hz, 2H), 7.43 (t, J = 7.3 Hz, 2H), 4.57 (t, J = 6.7 Hz, 1H), 4.33 (t, J = 14.5 Hz, 1H), 4.14 (s, 1H), 4.06 (t, J = 5.7 Hz, 2H), 3.83 (s, 3H), 3.62 (s, 1H), 3.58 (s, 2H), 3.34 (d, J = 6.6 Hz, 2H), 3.14 (s, 1H), 2.63 (dp, J = 5.5, 1.8 Hz, 2H), 2.60 (d, J = 2.0 Hz, 3H), 2.54 (d, J = 2.7 Hz, 1H), 2.42 (d, J = 0.8 Hz, 3H), 2.36 (p, J = 1.9 Hz, 1H), 2.00 (d, J = 7.6 Hz, 1H), 1.76 (d, J = 13.0 Hz, 1H), 1.68 (d, J = 4.8 Hz, 1H), 1.63 (d, J = 0.9 Hz, 3H); MS m / z [M+H] + 769.43.

[0967] [Production Example 5-26']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thie no[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)butanamido)benzoate hydrochloride

[0968] TIFF2025503429000431.tif31170

[0969] Step 1: methyl 4-aminobutanoate (intermediate 5-26a)

[0970] TIFF2025503429000432.tif15170

[0971] 4-Aminobutanoic acid (14.0 mmol, 1.0 eq) was dissolved in 20.0 mL of MeOH, and the gas in the reaction vessel was replaced with Ar gas. 1.7 mL of SOCl2 was added dropwise to the reaction mixture at 0 °C, and the mixture was stirred overnight at room temperature. The reaction mixture was distilled under reduced pressure and recrystallized from EtOAc to synthesize intermediate 5-26a.

[0972] Step 2: (S)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)butanoic acid (Intermediate 5-26'b)

[0973] TIFF2025503429000433.tif31170

[0974] Intermediate 5-26'b was synthesized from Intermediate 5-25'b and Intermediate 5-26a in the same manner as in Steps 4 and 5 of Production Example 5-25'.

[0975] ステップ3:methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(1-(2-(4-(4-chlorophenyl))-2,3,9-trimethyl-6H-thieno [3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)butanamido)benzoate hydrochloride

[0976] Intermediate 5-26'b and Intermediate 3-25a were synthesized by the same method as Production Example 5-7' and Example 5-26'.

[0977] 1 H NMR (500 MHz, DMSO-d6) δ 10.50 (s, 1H), 8.39 (s, 3H), 8.08 (s, 1H), 7.95 - 7.88 (m, 1H), 7.84 (d, J = 8.6 Hz, 1H), 7.59 - 7.53 (m, 1H), 7.50 - 7.38 (m, 4H), 4.59 - 4.52 (m, 1H), 4.30 (s, 2H), 4.02 (s, 2H), 3.77 (s, 3H), 3.63 - 3.47 (m, 1H), 3.43 - 3.30 (m, 1H), 3.16 - 3.01 (m, 4H), 2.62 - 2.56 (m, 3H), 2.37 (s, 3H), 1.67 (s, 4H), 1.57 (s, 3H), 1.32 (dd, J = 10.4, 6.7 Hz, 2H) 1.26 - 1.13 (m, 2H), 1.09 - 0.99 (m, 2H); MS m / z [M+H] + 783.31.

[0978] [Production Example 5-27']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-((3-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[ 3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)propyl)carbamoyl)benzoate hydrochloride

[0979] TIFF2025503429000434.tif31170

[0980] Step 1: (S)-N-(3-aminopropyl)-1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamide trifluoroacetic acid (intermediate 5-27'b)

[0981] TIFF2025503429000435.tif31170

[0982] Intermediate 5-27'b was synthesized from intermediate 5-25'b and tert-butyl(3-aminopropyl)carbamate in the same manner as in steps 1 and 2 of Preparation Example 5-140'.

[0983] Step 2: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-((3-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3, 2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)propyl)carbamoyl)benzoate hydrochloride

[0984] Example 5-27' was synthesized from intermediate 5-27'b, 3-iodo-4-(methoxycarbonyl)benzoic acid, in the same manner as in Step 5 of Example 5-7' and Example 1-1.

[0985] 1 H NMR (500 MHz, DMSO-d6) δ 8.76 (d, J = 3.9 Hz, 1H), 8.37 (s, 3H), 8.02 (d, J = 1.9 Hz, 1H), 7.97 - 7.85 (m, 3H), 7.45 (d, J = 8.7 Hz, 2H), 7.40 (dd, J = 12.3, 8.5 Hz, 2H), 4.54 (t, J = 6.7 Hz, 1H), 4.32 (s, 1H), 4.11 (s, 1H), 4.02 (q, J = 5.6 Hz, 2H), 3.85 (s, 3H), 3.58 - 3.47 (m, 4H), 3.25 (d, J = 6.0 Hz, 2H), 3.12 - 3.00 (m, 4H), 2.56 (s, 3H), 2.38 (s, 3H), 1.77 - 1.71 (m, 1H), 1.63 (d, J = 3.7 Hz, 2H), 1.59 (s, 3H), 1.41 - 1.30 (m. 1H), 1.25 - 1.18 (m, 1H); MS m / z [M+H] + 783.61.

[0986] [Production Example 5-28']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(3-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno [3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)prop-1-yn-1-yl)benzoate hydrochloride

[0987] TIFF2025503429000436.tif37170

[0988] Example 5-28' was synthesized from Intermediate 5-25'b and Intermediate 5-116c in the same manner as in Preparation Example 5-7'.

[0989] 1 H NMR (500 MHz, DMSO-d6) δ 8.45 (s, 4H), 7.86 (dd, J = 8.2, 2.8 Hz, 1H), 7.57 (t, J = 2.0 Hz, 1H), 7.55 - 7.50 (m, 1H), 7.49 - 7.45 (m, 3H), 7.41 (dd, J = 12.6, 8.4 Hz, 1H), 4.57 (t, J = 6.7 Hz, 1H), 4.33 (t, J = 13.2 Hz, 1H), 4.13 (dd, J = 7.7, 5.6 Hz, 4H), 4.00 (t, J = 5.7Hz, 4H), 3.83 (s, 4H), 3.62 - 3.37 (m, 3H), 3.13 (q, J = 11.1, 9.5 Hz, 1H), 2.59 (s, 3H), 2.40 - 2.38 (m, 3H), 1.59 (s, 3H); MS m / z [M+H] + 736.46.

[0990] [Production Example 5-29']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thie no[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)methyl)piperazin-1-yl)benzoate hydrochloride

[0991] TIFF2025503429000437.tif39170

[0992] Example 5-29' was synthesized from JQ1 carboxylic acid and intermediate 5-29d in the same manner as in Preparation Example 5-7'.

[0993] 1 H NMR (500 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.31 (s, 3H), 7.85 (d, J = 8.9 Hz, 1H), 7.53 - 7.47 (m, 1H), 7.44 (dd, J = 8.5, 2.6 Hz, 2H), 7.15 (d, J = 9.0 Hz, 1H), 7.10 (s, 1H), 4.58 (t, J = 6.7 Hz, 1H), 4.38 (d, J = 12.7 Hz, 1H), 4.20 (s, 1H), 4.04 (dd, J = 10.6, 6.3 Hz, 1H), 3.81 (s, 2H), 3.65 (s, MS m / z [M+H] + 753.54.

[0994] [Production Example 5-39']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(2-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[ 3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)-2,7-diazaspiro[3.5]nonane-7-carbonyl)benzoate trifluoroacetic acid

[0995] TIFF2025503429000438.tif29170

[0996] Example 5-39' was synthesized from intermediate 5-39c in the same manner as in steps 4 and 5 of Preparation Example 5-85'.

[0997] 1H NMR (500 MHz, DMSO-d6) δ 8.29 (s, 3H), 7.94 (d, J = 8.0 Hz, 1H), 7.56 (d, J = 1.7 Hz, 1H), 7.52 (dd, J = 8.0, 1.7 Hz, 1H), 7.45 (d, J = 9.6 Hz, 2H), 7.39 (s, 2H), 4.03 (s, 3H), 3.85 (s, 3H), 3.61 (s, 2H), 3.22 (t, J = 14.2 Hz, 4H), 3.10 (d, J = 21.5 Hz, 1H), 2.56 (s, 3H), 2.37 (s, 3H), 1.79 (s, 2H), 1.69 (s, 2H), 1.58 (s, 3H); MS m / z [M+H] + 724.47.

[0998] [Production Example 5-40']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(6-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thie no[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)-2,6-diazaspiro[3.3]heptan-2-yl)benzoate trifluoroacetic acid

[0999] TIFF2025503429000439.tif39170

[1000] Example 5-40' was synthesized from Intermediate 5-40b in the same manner as in Steps 4 and 5 of Preparation Example 5-85'.

[1001] 1H NMR (500 MHz, DMSO-d6) δ 8.26 (s, 3H), 7.76 (s, 1H), 7.47 (d, J = 8.6 Hz, 2H), 7.39 (d, J = 8.2 Hz, 2H), 6.54 - 6.43 (m, 2H), 4.59 - 4.40 (m, 3H), 4.15 - 3.96 (m, 8H), 3.73 (s, 3H), 3.22 (dd, J = 15.5, 7.2 Hz, 1H), 3.09 (dd, J = 15.8, 7.0 Hz, 1H), 2.56 (s, 3H), 2.37 (s, 3H), 1.58 (s, 3H); MS m / z [M+H] + 668.53.

[1002] [Production Example 5-41']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(6-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[ 3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)-2,6-diazaspiro[3.3]heptane-2-carbonyl)benzoate trifluoroacetic acid

[1003] TIFF2025503429000440.tif31170

[1004] Example 5-41' was synthesized from Intermediate 5-41c in the same manner as in Steps 4 and 5 of Preparation Example 5-85'.

[1005] 11H NMR (500 MHz, DMSO-d6) δ 8.29 (s, 3H), 7.94 (dd, J = 8.3, 4.8 Hz, 1H), 7.80 (s, 1H), 7.73 (d, J = 8.2 Hz, 1H), 7.44 (dd, J = 18.8, 8.3 Hz, 2H), 7.41 - 7.36 (m, 2H), 4.55 - 4.40 (m, 3H), 4.25 (s, 8H), 4.07 - 4.00 (m, 3H), 3.85 (s, 3H), 3.22 - 3.13 (m, 1H), 3.13 - 3.03 (m, 1H), 2.56 (s, 3H), 2.37 (s, 3H), 1.57 (s, 3H); MS m / z [M+H] + 696.60.

[1006] [Production Example 5-44'] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(5-(2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)benzoate trifluoroacetic acid

[1007] TIFF2025503429000441.tif37170

[1008] Step 1: tert-butyl 5-(3-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(methoxycarbonyl)phenyl)hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (Intermediate 5-44a)

[1009] TIFF2025503429000442.tif32170

[1010] Intermediate 5-44a was synthesized in the same manner as in Step 1 of Preparation Example 5-168, using intermediate 5-35a, tert-butyl hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate as a starting material.

[1011] Step 2: methyl 2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)benzoate hydrochloride (Intermediate 5-44b)

[1012] TIFF2025503429000443.tif32170

[1013] Intermediate 5-44b was synthesized using Intermediate 5-44a as a starting material in the same manner as in Step 2 of Production Example 1-1.

[1014] Step 3: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(5-(2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1 ,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)benzoate trifluoroacetic acid

[1015] Example 5-44' was synthesized using Intermediate 5-44b as a starting material in the same manner as in Steps 4 and 5 of Preparation Example 5-85'.

[1016] 1H NMR (500 MHz, DMSO-d6) δ 8.26 (s, 3H), 7.82 - 7.74 (m, 1H), 7.49 - 7.44 (m, 1H), 7.42 - 7.37 (m, 2H), 7.37 - 7.32 (m, 2H), 6.63 - 6.58 (m, 1H), 4.52 - 4.49 (m, 1H), 4.00 (s, 2H), 3.74 (d, J = 2.0 Hz, 3H), 3.58 (td, J = 13.3, 12.0, 6.4 Hz, 4H), 3.34 - 3.28 (m, 2H), 3.21 (h, J = 5.6 Hz, 2H), 3.15 (s, 1H), 3.03 (s, 1H), 2.55 (d, J = 1.0 Hz, 3H), 2.38 - 2.36 (m, 3H), 1.60 - 1.58 (m, 3H); MS m / z [M+H] + 682.41.

[1017] [Production Example 5-85'] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-(((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)methyl)amino)benzoate trifluoroacetic acid

[1018] TIFF2025503429000444.tif36170

[1019] Step 1: tert-butyl 4-(((3-iodo-4-(methoxycarbonyl)phenyl)amino)methyl)piperidine-1-carboxylate (Intermediate 5-85a)

[1020] TIFF2025503429000445.tif12170

[1021] Methyl 4-amino-2-iodobenzoate (0.9 mmol, 1.0 eq) and boc-piperidine carboxaldehyde (1.2 eq) were dissolved in dry DCM and stirred at room temperature for 15 minutes. Sodium triacetoxyborohydride (1.4 eq) was added and stirred at room temperature for 15 hours. The reaction mixture was extracted with DCM and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and purified via flash column to synthesize intermediate 5-85a.

[1022] Step 2: tert-butyl 4-(((3-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(methoxycarbonyl)phenyl)amino)methyl)piperidine-1-carboxylate (Intermediate 5-85b)

[1023] TIFF2025503429000446.tif26170

[1024] Intermediate 5-85a (0.2 mmol, 1.0 eq), n-propargylphthalimide (1.5 eq), Pd(PPh3)4 (0.01 eq), CuI (0.03 eq), TEA (0.2 mL), and DMF (0.8 mL) were mixed, and the gas in the reaction vessel was replaced with Ar gas. The mixture was then stirred overnight at room temperature. The reaction mixture was extracted with DCM and HO, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column to yield intermediate 5-85b.

[1025] Step 3: Methyl 2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-((piperidin-4-ylmethyl)amino)benzoate trifluoroacetic acid (Intermediate 5-85c)

[1026] TIFF2025503429000447.tif28170

[1027] Intermediate 5-85c was synthesized from intermediate 5-85b in the same manner as in Step 4 of Production Example 4-1′.

[1028] Step 4: methyl(S)-4-(((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diaze pin-6-yl)acetyl)piperidin-4-yl)methyl)amino)-2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoate (intermediate 5-85'a)

[1029] TIFF2025503429000448.tif39170

[1030] Intermediate 5-85'a was synthesized from intermediate 5-85c in the same manner as in Step 5 of Preparation Example 4-1'.

[1031] Step 5: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thi eno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)methyl)amino)benzoate trifluoroacetic acid

[1032] Intermediate 5-85'a (0.1 mmol, 1.0 eq), hydrazine monohydrate (3.0 eq), and 1.0 mL of EtOH were stirred for 16 hours at 40° C. The reaction product was distilled under reduced pressure and then purified by prep HPLC to synthesize Example 5-85'.

[1033] 11H NMR (500 MHz, DMSO-d6) δ 8.27 (s, 3H), 7.72 (dd, J = 8.7, 2.0 Hz, 1H), 7.49 (dd, J = 8.9, 1.8 Hz, 2H), 7.43 (dd, J = 8.6, 6.6 Hz, 2H), 6.82 (s, 1H), 6.71 - 6.65 (m, 2H), 4.58 (t, J = 6.7 Hz, 1H), 4.39 (d, J = 12.9 Hz, 1H), 4.18 (d, J = 13.6 Hz, 1H), 4.03 (q, J = 5.6 Hz, 2H), 3.75 (s, 3H), 3.69 - 3.58 (m, 2H), 3.12 (s, 2H), 3.00 (s, 2H), 2.63 (ddt, J = 7.5, 3.7, 1.9 Hz, 2H), 2.60 (d, J = 2.6 Hz, 3H), 2.42 (d, J = 1.0 Hz, 3H), 1.85 (s, 2H), 1.76 (s, 1H), 1.63 (s, 3H); MS m / z [M+H] + 684.51.

[1034] [Production Example 5-86’] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)amino)benzoate

[1035] TIFF2025503429000449.tif31170

[1036] Step 1: methyl(S)-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]di azepin-6-yl)acetyl)piperidin-4-yl)amino)-2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoate (intermediate 5-86'a)

[1037] TIFF2025503429000450.tif32170

[1038] Intermediate 5-86'a was synthesized from intermediate 5-86c in the same manner as in Step 5 of Preparation Example 4-1'.

[1039] Step 2: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)amino)benzoate

[1040] Example 5-86' was synthesized from Intermediate 5-86'a in the same manner as in Step 3 of Preparation Example 5-116.

[1041] 11H NMR (500 MHz, DMSO-d6) δ 10.48 (s, 1H), 8.36 (s, 3H), 8.28 (t, J = 5.7 Hz, 1H), 8.19 (d, J = 2.2 Hz, 1H), 7.71 (d, J = 8.8 Hz, 1H), 7.58 (d, J = 9.2 Hz, 1H), 7.39 (q, J = 8.8 Hz, 5H), 4.47 (dd, J = 8.1, 6.2 Hz, 1H), 4.04 (q, J = 5.6 Hz, 2H), 3.23 - 3.10 (m, 4H), 2.56 (s, 3H), 2.41 - 2.39 (m, 2H), 2.37 (d, J = 0.9 Hz, 3H), 1.74 (q, J = 7.2 Hz, 2H), 1.57 (d, J = 0.9 Hz, 3H); MS m / z [M+H] + 670.53.

[1042] [Production Example 5-139'] (S)-2-(3-aminoprop-1-yn-1-yl)-4-(((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)methyl)amino)benzoic acid trifluoroacetic acid

[1043] TIFF2025503429000451.tif36170

[1044] Step 1: (S)-4-(((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)methyl)amino)-2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoic acid (Intermediate 5-139'a)

[1045] TIFF2025503429000452.tif37170

[1046] Intermediate 5-139'a was synthesized from intermediate 5-85'a in the same manner as in Step 1 of Preparation Example 3-2'.

[1047] Step 2: (S)-2-(3-aminoprop-1-yn-1-yl)-4-(((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thien o[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)methyl)amino)benzoic acid trifluoroacetic acid

[1048] Example 5-139' was synthesized from Intermediate 5-139'a in the same manner as in Step 5 of Preparation Example 5-85'.

[1049] 11H NMR (500 MHz, DMSO-d6) δ 8.31 (s, 3H), 7.84 - 7.79 (m, 1H), 7.49 (dd, J = 8.7, 2.8 Hz, 3H), 7.43 (dd, J = 8.7, 7.1 Hz, 3H), 7.17 (s, 1H), 6.86 (dd, J = 8.8, 2.3 Hz, 1H), 6.67 (d, J = 2.1 Hz, 1H), 6.58 (s, 1H), 4.61 - 4.54 (m, 1H), 4.40 (d, J = 12.9 Hz, 1H), 4.19 (d, J = 12.9 Hz, 1H), 3.92 (d, J = 5.8 Hz, 2H), 3.66 - 3.61 (m, 2H), 3.09 (s, 3H), 2.62 (d, J = 1.8 Hz, 2H), 2.60 (d, J = 3.5 Hz, 3H), 2.42 (d, J = 1.0 Hz, 3H), 1.89 - 1.76 (m, 3H), 1.63 (d, J = 0.9 Hz, 3H); MS m / z [M+H] + 670.53.

[1050] [Production Example 5-140'] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazine-1-carbonyl)benzoate hydrochloride

[1051] TIFF2025503429000453.tif34170

[1052] Step 1: tert-butyl(S)-4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazine-1-carboxylate (Intermediate 5-140a)

[1053] TIFF2025503429000454.tif36170

[1054] Intermediate 5-140'a was synthesized from tert-butyl piperazine-1-carboxylate in the same manner as in Step 5 of Preparation Example 4-1'.

[1055] Step 2: (S)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)-1-(piperazin-1-yl)ethane-1-one trifluoroacetic acid (intermediate 5-140'b)

[1056] TIFF2025503429000455.tif36170

[1057] Example 5-140'b was synthesized from Intermediate 5-140'a in the same manner as in Step 4 of Preparation Example 4-1'.

[1058] Step 3: methyl(S)-2-bromo-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazine-1-carbonyl)benzoate (Intermediate 5-140c)

[1059] TIFF2025503429000456.tif36170

[1060] Intermediate 5-140'c was synthesized from intermediate 5-140'b and 3-bromo-4-(methoxycarbonyl)benzoic acid in the same manner as in Step 5 of Preparation 4-1'.

[1061] Step 4: methyl(S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethy l-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazine-1-carbonyl)benzoate (intermediate 5-140'd)

[1062] TIFF2025503429000457.tif34170

[1063] Intermediate 5-140'd was synthesized from intermediate 5-140'c in the same manner as in Step 2 of Preparation Example 4-179'.

[1064] Step 5: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperazine-1-carbonyl)benzoate hydrochloride

[1065] Example 5-140' was synthesized from Intermediate 5-140'd in the same manner as in Step 2 of Preparation Example 1-1.

[1066] 11H NMR (500 MHz, DMSO-d6) δ 8.41 (s, 3H), 7.94 (s, 1H), 7.64 (s, 1H), 7.58 (d, J = 8.1 Hz, 1H), 7.46 (d, J = 8.3 Hz, 2H), 7.40 (d, J = 8.1 Hz, 2H), 4.56 (t, J = 6.7 Hz, 1H), 4.06 - 4.00 (m, 2H), 3.86 (s, 3H), 3.81 - 3.68 (m, 4H), 3.54 - 3.43 (m, 5H), 3.27 - 3.19 (m, 1H), 2.57 (s, 3H), 2.38 (s, 3H), 1.59 (s, 3H); MS m / z [M+H] + 684.51.

[1067] [Production Example 5-152'] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)-N-methylpiperidine-4-carboxamido)benzoate trifluoroacetic acid

[1068] TIFF2025503429000458.tif31170

[1069] Step 1: methyl (S)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)-2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoate (Intermediate 5-152'a)

[1070] TIFF2025503429000459.tif34170

[1071] Example 5-23' (0.04 mmol, 1.0 eq) and N-hydroxyphthalimide (1.0 eq) were dissolved in 2.0 mL of 0.1 M phosphate buffer (pH 7.0) and stirred at room temperature for 16 hours. The reaction mixture was extracted with DCM and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and then purified via flash column to synthesize intermediate 5-152'a.

[1072] Step 2: methyl(S)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepine- 6-yl)acetyl)-N-methylpiperidine-4-carboxamido)-2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)benzoate (intermediate 5-152'b)

[1073] TIFF2025503429000460.tif34170

[1074] Intermediate 5-152'b was synthesized from intermediate 5-152'a in the same manner as in Step 1 of Preparation Example 5-153'.

[1075] Step 3: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[ 3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)-N-methylpiperidine-4-carboxamido)benzoate trifluoroacetic acid

[1076] Example 5-152' was synthesized from intermediate 5-152'b in the same manner as in Step 5 of Preparation Example 5-85'.

[1077] 1 1H NMR (500 MHz, DMSO-d6) δ 7.96 (s, 1H), 7.57 (s, 1H), 7.52 - 7.44 (m, 4H), 7.39 (s, 1H), 4.57 - 4.51 (m, 1H), 4.32 - 4.31 (m, 1H), 4.09 (s, 2H), 3.87 (s, 3H), 3.72 - 3.56 (m, 4H), 3.44 (s, 3H), 2.58 (s, 3H), 2.39 (s, 3H), 1.77 - 1.67 (m, 1H), 1.60 (s, 3H), 1.42 - 1.31 (m, 1H), 1.00 - 0.95 (m, 1H), 0.89 - 0.80 (s, 1H); MS m / z [M+H] + 712.58.

[1078] [Production Example 5-153’] methyl (S)-2-(3-aminoprop-1-yn-1-yl)-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)(methyl)carbamoyl)benzoate trifluoroacetic acid

[1079] TIFF2025503429000461.tif31170

[1080] Step 1: methyl (S)-2-bromo-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)(methyl)carbamoyl)benzoate (Intermediate 5-153’a)

[1081] TIFF2025503429000462.tif29170

[1082] Intermediate 5-22'a (0.06 mmol, 1.0 eq) was dissolved in 0.5 mL of dry DMF, followed by the addition of NaH (1.3 eq) and MeI (5.0 eq) and stirring at room temperature for 2 hours. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, evaporated, and purified via flash column chromatography to give intermediate 5-153'a.

[1083] Step 2: methyl(S)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H- thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)(methyl)carbamoyl)benzoate (intermediate 5-153'b)

[1084] TIFF2025503429000463.tif29170

[1085] Intermediate 5-153'b was synthesized from intermediate 5-153'a in the same manner as in Step 2 of Preparation Example 4-179'.

[1086] Step 3: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-((1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno [3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidin-4-yl)(methyl)carbamoyl)benzoate trifluoroacetic acid

[1087] Intermediate 5-153'b (0.5 mmol, 1.0 eq) was dissolved in 3.0 mL of DCM, and then HCl solution (4.0 M in dioxane, excess) was added dropwise and stirred at room temperature for 2 hours. The reaction mixture was distilled under reduced pressure and purified by prep HPLC to synthesize Example 5-153'.

[1088] 1 H NMR (500 MHz, DMSO-d6) δ 8.30 (s, 3H), 7.98 (d, J = 8.1 Hz, 1H), 7.62 - 7.55 (m, 2H), 7.52 - 7.40 (m, 5H), 4.58 (s,2H), 4.07 (d, J = 5.6 Hz, 2H), 3.89 (s, 3H), 3.68 (d, J = 9.6 Hz, 2H), 3.51 (s, 3H), 2.89 (d, J = 13.5 Hz, 2H), 2.75 (s, 2H), 2.60 (s, 3H), 2.42 (s, 3H), 1.63 (s, 3H); MS m / z [M+H] + 712.48.

[1089] [Production Example 5-154']methyl(S)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4, 3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)-2-(3-(sulfamoylamino)prop-1-yn-1-yl)benzoate trifluoroacetic acid

[1090] TIFF2025503429000464.tif41170

[1091] To a mixture of Example 5-23' (0.043 mmol, 1.0 eq), TEA (2.5 eq), and 0.8 mL of DCM, sulfamoyl chloride (0.048 mmol, 1.1 eq) dissolved in 0.2 mL of DCM was added dropwise at 0 °C and stirred overnight at room temperature. The reaction mixture was extracted with EtOAc and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and purified by prep HPLC to synthesize Example 5-154'.

[1092] 1 H NMR (500 MHz, DMSO-d6) δ 10.36 (d, J = 15.9 Hz, 1H), 7.95 (d, J = 2.1 Hz, 1H), 7.87 (dd, J = 8.7, 3.7 Hz, 1H), 7.68 - 7.60 (m, 1H), 7.51 - 7.43 (m, 4H), 7.14 - 7.11 (m, 1H), 6.67 (s, 3H), 4.59 (t, J = 6.7 Hz, 1H), 4.45 - 4.39 (m, 1H), 4.25 - 4.20 (m, 1H), 3.97 (d, J = 5.7 Hz, 2H), 3.82 (s, 3H), 3.65 - 3.56 (m, 1H), 3.22 - 3.20 (m, 1H), 2.73 - 2.62 (m, 3H), 2.60 (s, 3H), 2.42 (s, 3H), 1.96 - 1.92 (m, 1H), 1.87 - 1.81 (m, 1H), 1.73 - 1.66 (m, 1H), 1.64 (s, 3H), 1.47 (t, J = 14.9 Hz, 1H), MS m / z [M+H] + 777.32,

[1093] [Production Example 5-155']methyl(S)-2-((1-aminocyclopropyl)ethynyl)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl- 6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)benzoate trifluoroacetic acid

[1094] TIFF2025503429000465.tif37170

[1095] Step 1: methyl 4-amino-2-((1-((tert-butoxycarbonyl)amino)cyclopropyl)ethynyl)benzoate (Intermediate 5-155a)

[1096] TIFF2025503429000466.tif26170

[1097] Example 5-155a was synthesized from methyl 4-amino-2-iodobenzoate and tert-butyl(1-ethynylcyclopropyl)carbamate in the same manner as in Preparation Example 1-1.

[1098] Step 2: methyl(S)-2-((1-((tert-butoxycarbonyl)amino)cyclopropyl)ethynyl)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimeth yl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)benzoate (intermediate 5-155'a)

[1099] TIFF2025503429000467.tif31170

[1100] Intermediate 5-155'a was synthesized from Intermediate 5-25'b and Intermediate 5-155a in the same manner as in Step 5 of Preparation Example 4-1'.

[1101] Step 3: methyl(S)-2-((1-aminocyclopropyl)ethynyl)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)benzoate trifluoroacetic acid

[1102] Example 5-155' was synthesized from Intermediate 5-155'a in the same manner as in Step 3 of Preparation Example 5-153'.

[1103] 11H NMR (500 MHz, DMSO-d6) δ 10.43 (d, J = 10.9 Hz, 1H), 8.77 (s, 2H), 8.14 (dd, J = 22.6, 2.2 Hz, 1H), 7.92 (dd, J = 8.7, 3.5 Hz, 1H), 7.59 - 7.48 (m, 3H), 7.45 (t, J = 8.6 Hz, 2H), 4.60 (d, J = 6.8 Hz, 1H), 3.82 (d, J = 2.3 Hz, 3H), 3.61 (dd, J = 16.3, 7.5 Hz, 1H), 3.44 - 3.36 (m, 1H), 3.22 (q, J = 11.2 Hz, 1H), 2.69 (t, J = 12.1 Hz, 2H), 2.61 (s, 3H), 2.42 (d, J = 1.0 Hz, 3H), 1.94 (d, J = 12.6 Hz, 1H), 1.83 (s, 1H), 1.74 - 1.66 (m, 1H), 1.64 (s, 3H), 1.48 (d, J = 15.8 Hz, 1H), 1.42 - 1.38 (m, 2H), 1.36 - 1.33 (m, 2H), 1.26 - 1.21 (m, 2H); MS m / z [M+H] + 724.47.

[1104] [Production Example 5-157'] methyl 4-(1-(2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)-2-(3-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)prop-1-yn-1-yl)benzoate

[1105] TIFF2025503429000468.tif43170

[1106] To a mixture of Example 5-23' (0.027 mmol, 1.0 eq), HOBt (1.0 eq), DIPEA (3.0 eq), and 0.5 mL of DMF, 1-(((4-nitrophenoxy)carbonyl)oxy)ethyl isobutyrate (1.0 eq) was added at 0°C and stirred for 20 minutes. The reaction product was filtered under reduced pressure, purified by flash column, and further purified by prep HPLC to synthesize Example 5-157'.

[1107] 1 H NMR (500 MHz, DMSO-d6) δ 10.33 (d, J = 16.3 Hz, 1H), 8.07 - 8.01 (m, 1H), 7.93 (dd, J = 6.5, 2.2 Hz, 1H), 7.86 (dd, J = 8.7, 3.6 Hz, 1H), 7.63 (dt, J = 8.7, 2.5 Hz, 1H), 7.50 (d, J = 8.4 Hz, 2H), 7.48 - 7.41 (m, 2H), 6.70 (q, J = 5.4 Hz, 1H), 4.59 (t, J = 6.7 Hz, 1H), 4.46 - 4.38 (m, 1H), 4.27 - 4.19 (m, 1H), 4.13 - 4.02 (m, 2H), 3.80 (d, J = 1.2 Hz, 3H), 3.22 (q, J = 11.6 Hz, 1H), 2.75 - 2.64 (m, 2H), 2.61 (s, 3H), 2.42 (s, 3H), 1.97 - 1.91 (m, 1H), 1.87 - 1.80 (m, 1H), 1.74 - 1.67 (m, 1H), 1.64 (s, 3H), 1.52 - 1.44 (m, 1H), 1.41 (d, J = 5.4 Hz, 3H), 1.06 (dd, J = 7.0, 4.5 Hz, 6H); MS m / z [M+H] + 856.44

[1108] [Production Example 5-158']methyl(S)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4 ,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)-2-(3-(sulfamoyloxy)prop-1-yn-1-yl)benzoate trifluoroacetic acid

[1109] TIFF2025503429000469.tif45170

[1110] Step 1: tert-butyl 4-((3-iodo-4-(methoxycarbonyl)phenyl)carbamoyl)piperidine-1-carboxylate (Intermediate 5-158a)

[1111] TIFF2025503429000470.tif15170

[1112] Intermediate 5-158'a was synthesized in the same manner as in Step 5 of Preparation Example 4-1' using 1-(tert-Butoxycarbonyl)piperidine-4-carboxylic acid and methyl 4-amino-2-iodobenzoate as starting materials.

[1113] Step 2: methyl 2-iodo-4-(piperidine-4-carboxamido)benzoate dihydrochloride (Intermediate 5-158b)

[1114] TIFF2025503429000471.tif14170

[1115] Intermediate 5-158'b was synthesized from Intermediate 5-158'a in the same manner as in Step 2 of Production Example 1-1.

[1116] Step 3: methyl(S)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)-2-iodobenzoate (intermediate 5-158'c)

[1117] TIFF2025503429000472.tif32170

[1118] Intermediate 5-158'c was synthesized from JQ1 carboxylic acid and intermediate 5-158'b in the same manner as in Step 5 of Preparation Example 4-1'.

[1119] Step 4: methyl(S)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a] [1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)-2-(3-hydroxyprop-1-yn-1-yl)benzoate (intermediate 5-158'd)

[1120] TIFF2025503429000473.tif36170

[1121] Intermediate 5-158'c (0.3 mmol, 1 eq), propargyl alcohol (3.0 eq), Pd(PPh3)4 (0.02 eq), CuI (0.1 eq), TEA (1.0 mL), and DMF (1.0 mL) were mixed, and the gas in the reaction vessel was replaced with Ar gas. The mixture was stirred overnight at room temperature. The mixture was purified via flash column to synthesize intermediate 5-158'd.

[1122] Step 5: methyl(S)-4-(1-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3- a][1,4]diazepin-6-yl)acetyl)piperidine-4-carboxamido)-2-(3-(sulfamoyloxy)prop-1-yn-1-yl)benzoate trifluoroacetic acid

[1123] Example 5-158' was synthesized from Intermediate 5-158'd in the same manner as in Preparation Example 5-154'.

[1124] 1 H NMR (500 MHz, DMSO-d6) δ 10.40 (d, J = 16.9 Hz, 1H), 8.00 (dd, J = 7.6, 2.2 Hz, 1H), 7.91 (dd, J = 8.7, 3.8 Hz, 1H), 7.72 (s, 2H), 7.69 (dd, J = 8.5, 2.5 Hz, 1H), 7.53 - 7.42 (m, 4H), 7.28 (s, 1H), 6.69 (s, 1H), 5.02 (s, 2H), 4.59 (t, J = 6.7 Hz, 1H), 4.42 (s, 1H), 4.23 (s, 1H), 3.82 (d, J = 1.1 Hz, 3H), 3.67 - 3.54 (m, 1H), 2.68 (s, 2H), 2.60 (s, 3H), 2.42 (s, 3H), 1.91 (s, 1H), 1.84 (s, 1H), 1.70 (d, J = 13.0 Hz, 1H), 1.64 (s, 3H), 1.47 (d, J = 6.3 Hz, 1H).

[1125] MS m / z [M+H] + 778.42.

[1126] [Production Example 5-162']methyl 4-(6-(2-((S)-4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)ac etyl)-2,6-diazaspiro[3.3]heptan-2-yl)-2-(3-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)prop-1-yn-1-yl)benzoate

[1127] TIFF2025503429000474.tif39170

[1128] Example 5-40 is the same as Manufacturing Example 5-157, and the same method is used in Example 5-162.

[1129] 1 H NMR (500 MHz, DMSO-d6) δ 7.98 (t, J = 5.8 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.50 - 7.45 (m, 2H), 7.42 - 7.37 (m, 2H), 6.66 (q, J = 5.4 Hz, 1H), 6.46 - 6.41 (m, 2H), 4.57 - 4.47 (m, 2H), 4.45 (t, J = 7.1 Hz, 1H), 4.11 (d, J = 3.1 Hz, 2H), 4.09 - 4.06 (m, 3H), 4.05 - 4.03 (m, 3H), 3.70 (s, 3H), 3.21 (dd, J = 15.5, 6.9 Hz, 1H), 3.11 (dd, J = 15.5, 7.2 Hz, 1H), 2.57 (s, 3H), 2.37 (d, J = 1.1 Hz, 3H), 1.59 (d, J = 1.0 Hz, 3H), 1.37 (d, J = 5.4 Hz, 3H), 1.03 (dd, J = 7.0, 3.9 Hz, 6H).

[1130] [Production Example 5-168']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(7-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thi eno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetyl)-2,7-diazaspiro[3.5]nonan-2-yl)benzoate hydrochloride

[1131] TIFF2025503429000475.tif37170

[1132] Preparation 5-168′ was synthesized from JQ1 carboxylic acid and intermediate 5-168b in the same manner as in steps 4 and 5 of Preparation 5-85′.

[1133] 1 H NMR (400 MHz, DMSO-d6) δ 7.76 (d, J = 2.3 Hz, 1H), 7.49 (s, 2H), 7.45 (s, 2H), 6.45 (s, 2H), 4.58 (t, J = 1.7 Hz, 1H), 3.74 (m, 6H), 3.64 (m, 5H), 3.48 (m, 2H), 3.28 (m, 2H), 2.60 (s, 3H), 2.42 (s, 3H), 1.88 (s, 2H), 1.70 (s, 2H), 1.63 (s, 3H); MS m / z [M+H] + 696.2.

[1134] [Manufacturing example IV'] A TBM-UBM compound in which a compound (JQ1) exhibiting activity as a BRD4 (bromodomain-containing protein 4) inhibitor is linked to a compound corresponding to Formula IV of the present invention was synthesized as follows.

[1135] [Production Example 5-32']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(3-(2-(4-(4-chlorophenyl))-2,3,9-trimethyl-6H- thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanoyl)piperazin-1-yl)benzoate hydrochloride

[1136] TIFF2025503429000476.tif31170

[1137] Intermediate 4-124'a and Intermediate 5-7b are synthesized by the same method as Production Example 5-7' and Example 5-32'.

[1138] 1 H NMR (500 MHz, DMSO-d6) δ 8.34 (s, 3H), 8.24 (t, J = 5.8 Hz, 1H), 7.78 (d, J = 8.9 Hz, 1H), 7.46 - 7.43 (m, 2H), 7.39 (d, J = 8.6 Hz, 2H), 7.00 (dd, J = 9.0, 2.7 Hz, 1H), 6.97 (d, J = 2.6 Hz, 1H), 4.47 (t, J = 7.1 Hz, 1H), 3.98 (q, J = 5.6 Hz, 2H), 3.75 (s, 3H), 3.59 - 3.54 (m, 3H), 3.39 - 3.26 (m, 9H), 3.19 (d, J = 7.2 Hz, 2H), 2.55 (s, 3H), 2.37 (s, 3H), 1.58 (s, 3H); MS m / z [M+H] + 727.47.

[1139] [Production Example 5-33']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-5-(4-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H- thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanoyl)piperazin-1-yl)benzoate hydrochloride

[1140] TIFF2025503429000477.tif31170

[1141] Step 1: methyl 5-bromo-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate (Intermediate 5-33a)

[1142] TIFF2025503429000478.tif18170

[1143] Intermediate 5-33a was synthesized from methyl 5-bromo-2-iodobenzoate in the same manner as in Step 1 of Production Example 1-1.

[1144] Step 2: methyl 2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-5-(piperazin-1-yl)benzoate (Intermediate 5-33b)

[1145] TIFF2025503429000479.tif18170

[1146] Intermediate 5-33b was synthesized from intermediate 5-33a and piperazine in the same manner as in Step 1 of Preparation Example 5-40.

[1147] Step 3: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-5-(4-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-th ieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propanoyl)piperazin-1-yl)benzoate hydrochloride

[1148] Example 5-33' was synthesized from Intermediate 4-124'a and Intermediate 5-33b in the same manner as in Preparation Example 1-1'.

[1149] 1 H NMR (500 MHz, DMSO-d6) δ 8.26 (d, J = 26.6 Hz, 3H), 7.45 (dd, J = 8.9, 1.7 Hz, 2H), 7.43 - 7.36 (m, 3H), 7.29 (d, J = 2.8 Hz, 1H), 7.13 (dd, J = 8.8, 2.6 Hz, 1H), 7.07 (s, 1H), 4.48 (t, J = 7.1 Hz, 1H), 3.95 (q, J = 5.6 Hz, 1H), 3.81 (s, 3H), 3.36 - 3.27 (m, 5H), 3.27 - 3.18 (m, 5H), 2.56 (d, J = MS m / z [M+H] + 727.47.

[1150] [Production Example 5-111'](S)-5-(4-(3-aminoprop-1-yn-1-yl)phenyl)-N-(7-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl -6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)heptyl)furan-2-carboxamide hydrochloride

[1151] TIFF2025503429000480.tif29170

[1152] Step 1: (S)-N-(7-aminoheptyl)-2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamide hydrochloride (Intermediate 5-111a)

[1153] TIFF2025503429000481.tif31170

[1154] Intermediate 5-111'a was synthesized from tert-butyl(7-aminopropyl)carbamate in the same manner as in Production Example 1-1'.

[1155] Step 2: (S)-5-(4-bromophenyl)-N-(7-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)heptyl)furan-2-carboxamide (Intermediate 5-111b)

[1156] TIFF2025503429000482.tif29170

[1157] Intermediate 5-111'b was synthesized from intermediate 5-111'a and 5-(4-Bromophenyl)-2-furoic acid in the same manner as in Step 5 of Preparation Example 4-1'.

[1158] ステップ3:(S)-5-(4-(3-aminoprop-1-yn-1-yl)phenyl)-N-(7-(2-(4-(4-chlorophenyl))-2,3,9-trimethyl-6 H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)heptyl)furan-2-carboxamide hydrochloride

[1159] Intermediate 5-111'b is produced by the same method as in Production Example 4-179' of Preventive Materials 2~3, and Example 5-111' is synthesized by the same method.

[1160] 1 H NMR (500 MHz, DMSO-d6) δ 8.48 (t, J = 5.8 Hz, 1H), 8.14 (t, J = 5.7 Hz, 1H), 7.87 - 7.83 (m, 2H), 7.46 - 7.41 (m, 5H), 7.39 - 7.36 (m, 2H), 7.09 (q, J = 3.5 Hz, 2H), 4.46 (dd, J = 8.1, 6.1 Hz, 1H), 3.49 (s, 2H), 3.19 (d, J = 5.8 Hz, 3H), 3.14 - 3.09 (m, 2H), 3.05 (dq, J = 12.9, 6.2 Hz, 2H), 2.54 (d, J = 2.0 Hz, 3H), 2.35 (d, J = 3.1 Hz, 3H), 1.57 (s, 3H), 1.52 (d, J = 8.1 Hz, 2H), 1.48 (d, J = 7.6 Hz, 2H), 1.40 (s, 2H), 1.19 (s, 4H); MS m / z [M+H] + 736.56.

[1161] [Production Example 5-164']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6 H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)piperazin-1-yl)benzoate hydrochloride

[1162] TIFF2025503429000483.tif39170

[1163] Step 1: methyl 4-(4-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propyl)piperazin-1-yl)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate (intermediate 5-164a)

[1164] TIFF2025503429000484.tif26170

[1165] Intermediate 5-7b (0.1 mmol, 1.0 eq) was dissolved in 3.0 mL of DCM, and then NaBH(OAc) (3.0 eq), (9H-fluoren-9-yl)methyl(3-oxopropyl)carbamate (1.2 eq), and acetic acid (1.0 eq) were slowly added at 0 °C and stirred for 12 hours. The reaction mixture was extracted with DCM and HO, and the organic layer was dried over NaSO. The mixture was filtered under reduced pressure, distilled, and purified via flash column to yield intermediate 5-164a.

[1166] Step 2: methyl 4-(4-(3-aminopropyl)piperazin-1-yl)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate formic acid (intermediate 5-164b)

[1167] TIFF2025503429000485.tif30170

[1168] Intermediate 5-164a (0.1 mmol, 1.0 eq) was dissolved in 2.0 mL of piperidine / DMF (1:1) and stirred at room temperature for 0.5 hours. The reaction mixture was concentrated under reduced pressure and purified by prep HPLC to synthesize intermediate 5-164b.

[1169] Step 3: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-4-(4-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-t hieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)piperazin-1-yl)benzoate hydrochloride

[1170] Example 5-164' was synthesized from JQ1 carboxylic acid and intermediate 5-164b in the same manner as in Step 2 of Preparation Example 5-77'.

[1171] 1 H NMR (400 MHz, DMSO-d6) δ 11.37 (s, 1H), 8.65 - 8.47 (m, 3H), 7.84 (d, J = 2.2 Hz, 1H), 7.53 (d, J = 2.2 Hz, 2H), 7.44 (d, J = 2.1 Hz, 2H), 7.19 (s, 1H), 7.18 - 7.12 (m, 1H), 4.59 - 4.57 (m, 1H), 4.05 - 3.99 (m, 4H), 3.81 (s, 3H), 3.61 - 3.56 (m, 2H), 3.43 - 3.39 (m, 2H), 3.30 - 3.27 (m, 3H), 3.17 - 3.12 (m, 5H), 2.64 (s, 3H), 2.42 (m, 3H), 1.96 - 1.91 (m, 2H), 1.62 (s, 3H); MS m / z [M+H]+ 713.5.

[1172] [Production Example 5-165']methyl(S)-2-(3-aminoprop-1-yn-1-yl)-5-(4-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6 H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)piperazin-1-yl)benzoate hydrochloride

[1173] TIFF2025503429000486.tif34170

[1174] Step 1: methyl 5-(4-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propyl)piperazin-1-yl)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate (intermediate 5-165a)

[1175] TIFF2025503429000487.tif19170

[1176] Starting from 5-33b, intermediate 5-165a was synthesized in the same manner as in Step 1 of Preparation 5-164'.

[1177] Step 2: methyl 5-(4-(3-aminopropyl)piperazin-1-yl)-2-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)benzoate (intermediate 5-165b)

[1178] TIFF2025503429000488.tif19170

[1179] Starting from Intermediate 5-165a, Intermediate 5-165b was synthesized in the same manner as in Step 4 of Example 5-29.

[1180] Step 3: methyl(S)-2-(3-aminoprop-1-yn-1-yl)-5-(4-(3-(2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-t hieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetamido)propyl)piperazin-1-yl)benzoate hydrochloride

[1181] Starting from intermediate 5-165b, Example 5-165' was synthesized in the same manner as in Step 2 of Example 5-77'.

[1182] 1 H NMR (400 MHz, DMSO-d6) δ 8.21 (t, J = 1.4 Hz, 1H), 7.48 (d, J = 2.1 Hz, 2H), 7.42 (d, J = 2.1 Hz, 2H), 7.32 (d, J = 2.2 Hz, 1H), 7.25 (s, 1H), 7.11 (m, 1H), 4.50 (t, J = 1.7 Hz, 1H), 3.82 (s, 3H), 3.48 (s, 2H), 3.24 - 3.20 (m, 6H), 3.15 (m, 2H), 2.59 (s, 3H), 2.41 (s, 4H), 2.37 (s, 3H), 2.34 (m, 2H), 1.62 (s, 5H); MS m / z [M+H] + 713.4.

[1183] [Manufacturing example I''] A TBM-UBM compound in which a compound (alisertib) exhibiting activity as an Aurora A kinase inhibitor is linked to a compound corresponding to Formula I of the present invention was synthesized as follows.

[1184] [Production Example 1-1'']N-(4-(3-aminoprop-1-yn-1-yl)phenyl)-4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide trifluoroacetic acid

[1185] TIFF2025503429000489.tif37170

[1186] Example 1-1″ was synthesized from commercially available Alisertib (4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzoic acid) and intermediate 1-1a in the same manner as in steps 2 and 3 of Preparation Example 5-155′.

[1187] 1 H NMR (500 MHz, DMSO-d6) δ 10.23 (s, 1H), 10.10 (s, 1H), 8.71 (d, J = 3.6 Hz, 1H), 8.27 (d, J = 8.5 Hz, 1H), 8.22 (s, 3H), 7.98 (s, 1H), 7.79 (dd, J = 9.5, 6.9 Hz, 3H), 7.70 (d, J = 8.5 Hz, 1H), 7.41 (dddd, J = 12.3, 9.9, 8.5, 1.8 Hz, 4H), 3.98 - 3.94 (m, 2H), 3.92 (s, 3H); MS m / z [M+H] + 647.45.

[1188] [Production Example 1-3'']N-(3-(3-aminoprop-1-yn-1-yl)phenyl)-4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide trifluoroacetic acid

[1189] TIFF2025503429000490.tif40170

[1190] Example 1-3'' was synthesized from Alisertib and Intermediate 1-3a in the same manner as in Steps 2 and 3 of Preparation Example 5-155'.

[1191] 1 H NMR (500 MHz, DMSO-d6) δ 10.27 (s, 1H), 10.07 (s, 1H), 8.74 (s, 1H), 8.31 (d, J = 8.8 Hz, 4H), 8.11 (s, 1H), 8.01 (s, 1H), 7.83 (dd, J = 8.5, 2.2 Hz, 1H), 7.74 (d, J = 8.6 Hz, 1H), 7.66 - 7.63 (m, 1H), 7.48 (dd, J = 8.6, 1.9 Hz, 1H), 7.45 - 7.37 (m, 2H), 7.23 (s, 1H), 7.18 (d, J = 7.7 Hz, 1H), 4.03 (d, J = 5.6 Hz, 2H), 3.96 (s, 6H), 3.80 (s, 2H); MS m / z [M+H] + 647.45.

[1192] [Production Example 3-25'']methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)benzoate trifluoroacetic acid

[1193] TIFF2025503429000491.tif40170

[1194] Example 3-25'' was synthesized from Alisertib and intermediate 3-25a in the same manner as in steps 2 and 3 of Preparation Example 5-155'.

[1195] 1 H NMR (500 MHz, DMSO-d6) δ 10.30 (d, J = 4.6 Hz, 2H), 8.75 (s, 1H), 8.35 (s, 2H), 8.30 (d, J = 2.6 Hz, 3H), 8.02 (s, 1H), 7.93 (d, J = 8.6 Hz, 1H), 7.83 (dd, J = 8.5, 2.2 Hz, 1H), 7.73 (td, J = 6.4, 3.3 Hz, 2H), 7.48 (dd, J = 8.6, 1.9 Hz, 1H), 7.43 (td, J = 8.4, 6.8 Hz, 1H), 7.23 (s, 1H), 7.04 - 6.77 (m, 1H), 4.08 (s, 2H), 3.96 (s, 6H), 3.85 (s, 5H); MS m / z [M+H] + 705.39.

[1196] [Production Example 3-49'']N-(4-(3-amino-4-hydroxybut-1-yn-1-yl)phenyl)-4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide trifluoroacetic acid

[1197] TIFF2025503429000492.tif39170

[1198] Example 3-49'' was synthesized from Alisertib and intermediate 3-49b in the same manner as in steps 2 and 3 of Preparation Example 5-155'.

[1199] 1 H NMR (500 MHz, DMSO-d6) δ 10.23 (s, 1H), 10.10 (s, 1H), 8.70 (s, 1H), 8.40 (d, J = 5.3 Hz, 3H), 8.27 (d, J = 8.5 Hz, 1H), 7.97 (s, 1H), 7.82 - 7.76 (m, 4H), 7.70 (d, J = 8.6 Hz, 1H), 7.46 - 7.35 (m, 5H), 7.19 (s, 1H), 4.32 - 4.26 (m, 1H), 3.92 (s, 3H), 3.74 (dd, J = 11.3, 4.4 Hz, 1H), 3.63 (dd, J = 11.2, 7.0 Hz, 1H); MS m / z [M+H] + 677.52.

[1200] [Production Example 4-1'']N-(3-(5-(3-aminoprop-1-yn-1-yl)furan-2-yl)prop-2-yn-1-yl)-4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide trifluoroacetic acid

[1201] TIFF2025503429000493.tif40170

[1202] Example 4-1'' was synthesized from Alisertib using Intermediate 4-1d in the same manner as in Steps 4 and 5 of Preparation Example 5-85'.

[1203] 1H NMR (500 MHz, DMSO-d6) δ 10.22 (s, 1H), 8.70 (s, 1H), 8.52 (t, J = 5.7 Hz, 1H), 8.28 (s, 2H), 8.26 (s, 3H), 7.96 (s, 1H), 7.83 (d, J = 8.7 Hz, 1H), 7.79 (dd, J = 8.5, 2.2 Hz, 1H), 7.42 - 7.34 (m, 3H), 7.19 (s, 1H), 6.91 (d, J = 3.6 Hz, 1H), 6.84 (d, J = 3.6 Hz, 1H), 4.32 (d, J = 5.7 Hz, 2H), 4.04 (d, J = 5.5 Hz, 2H), 3.90 (s, 6H); MS m / z [M+H] + 675.42.

[1204] [Production Example 5-116'']methyl 2-(3-aminoprop-1-yn-1-yl)-4-(3-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)prop-1-yn-1-yl)benzoate trifluoroacetic acid

[1205] TIFF2025503429000494.tif40170

[1206] Example 5-116'' was synthesized from Alisertib and intermediate 5-116c in the same manner as in Steps 2 and 3 of Preparation Example 5-155'.

[1207] 1H NMR (500 MHz, DMSO-d6) δ 10.21 (d, J = 2.6 Hz, 1H), 8.70 (s, 1H), 8.52 (t, J = 5.7 Hz, 1H), 8.29 - 8.19 (m, 5H), 7.96 (s, 1H), 7.85 (dd, J = 11.4, 8.4 Hz, 2H), 7.79 (dd, J = 8.5, 2.3 Hz, 1H), 7.59 - 7.51 (m, 2H), 7.42 - 7.35 (m, 3H), 7.19 (s, 1H), 4.33 (d, J = 5.7 Hz, 2H), 4.01 (q, J = 5.6 Hz, 2H), 3.91 (s, 3H), 3.82 (s, 3H); MS m / z [M+H] + 743.45.

[1208] [Manufacturing example II''] A TBM-UBM compound in which a compound (alisertib) exhibiting activity as an Aurora A kinase inhibitor is linked to a compound corresponding to Formula II of the present invention was synthesized as follows.

[1209] [Production Example 4-165'']N-(6-((4-(3-aminoprop-1-yn-1-yl)phenyl)amino)-6-oxohexyl)-4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide trifluoroacetic acid

[1210] TIFF2025503429000495.tif38170

[1211] Step 1: methyl 6-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)hexanoate (Intermediate 4-165''a)

[1212] TIFF2025503429000496.tif41170

[1213] Intermediate 4-165''a was synthesized using alisertib and methyl 6-aminohexanoate as starting materials in the same manner as in Step 5 of Preparation Example 4-1'.

[1214] Step 2: Potassium 6-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)hexanoate (Intermediate 4-165''b)

[1215] TIFF2025503429000497.tif45170

[1216] Intermediate 4-165''a (0.04 mmol, 1.0 eq), potassium trimethylsilanolate (4.0 eq), and 0.5 mL of THF were mixed and stirred at room temperature for 3 hours. The reaction mixture was filtered under reduced pressure, and intermediate 4-165''b was synthesized without further purification.

[1217] Step 3: tert-butyl(3-(4-(6-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)hexanamido)phenyl)prop-2-yn-1-yl)carbamate (Intermediate 4-165''c)

[1218] TIFF2025503429000498.tif39170

[1219] Starting from intermediate 4-165″b and 1-1a, intermediate 4-165″c was synthesized in the same manner as in Step 5 of Preparation Example 4-1′.

[1220] Step 4: N-(6-((4-(3-aminoprop-1-yn-1-yl)phenyl)amino)-6-oxohexyl)-4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide trifluoroacetic acid

[1221] TIFF2025503429000499.tif42170

[1222] Example 4-165'' was synthesized from intermediate 4-165''c in the same manner as in Step 3 of Preparation 5-153'.

[1223] 1 H NMR (500 MHz, DMSO-d6) δ 10.15 (s, 1H), 10.07 (s, 1H), 8.68 (s, 1H), 8.26 (d, J = 8.6 Hz, 1H), 8.20 (s, 3H), 8.01 (s, 1H), 7.91 (s, 1H), 7.80 - 7.77 (m, 1H), 7.77 - 7.74 (m, 2H), 7.63 - 7.60 (m, 2H), 7.38 (d, J = 7.0 Hz, 1H), 7.37 - 7.34 (m, 4H), 3.94 (d, J = 5.7 Hz, 2H), 3.87 (s, 6H), 2.31 (d, J = 7.7 Hz, 4H), 1.60 (d, J = 7.4 Hz, 2H), 1.51 (t, J = 7.3 Hz, 2H), 1.32 (d, J = 7.2 Hz, 2H); MS m / z [M+H] + 760.44.

[1224] [Production Example 4-166'']5-(3-aminoprop-1-yn-1-yl)-N-(3-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)propyl)furan-2-carboxamide trifluoroacetic acid

[1225] TIFF2025503429000500.tif41170

[1226] Step 1: tert-butyl(3-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)propyl)carbamate (Intermediate 4-166''a)

[1227] TIFF2025503429000501.tif43170

[1228] Intermediate 4-166''a was synthesized in the same manner as in Step 1 of Preparation Example 4-165'' using tert-Butyl(3-aminopropyl)carbamate.

[1229] Step 2: N-(3-aminopropyl)-4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide hydrochloride (Intermediate 4-166''b)

[1230] TIFF2025503429000502.tif43170

[1231] Intermediate 4-166''b was synthesized from Intermediate 4-166''a in the same manner as in Step 2 of Preparation Example 1-1.

[1232] Step 3: tert-butyl(3-(5-((3-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azep in-2-yl)amino)-2-methoxybenzamido)propyl)carbamoyl)furan-2-yl)prop-2-yn-1-yl)carbamate (intermediate 4-166''c)

[1233] TIFF2025503429000503.tif45170

[1234] Intermediate 4-166''c was synthesized from Intermediate 4-166''b and Intermediate 4-227b in the same manner as in Step 1 of Production Example 1-1'.

[1235] Step 4: 5-(3-aminoprop-1-yn-1-yl)-N-(3-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)propyl)furan-2-carboxamide trifluoroacetic acid

[1236] Example 4-166'' was synthesized from intermediate 4-166''c in the same manner as in Step 3 of Preparation 5-153'.

[1237] 11H NMR (500 MHz, DMSO-d6) δ 10.20 (s, 1H), 8.72 (s, 1H), 8.60 (t, J = 6.0 Hz, 1H), 8.39 - 8.25 (m, 6H), 7.97 (s, 1H), 7.84 - 7.81 (m, 2H), 7.44 - 7.40 (m, 2H), 7.22 (s, 1H), 7.19 (d, J = 3.6 Hz, 1H), 7.01 (d, J = 3.6 Hz, 1H), 4.11 (d, J = 5.6 Hz, 2H), 3.94 (s, 6H), 3.30 (dt, J = 12.9, 6.4 Hz, 6H), 1.70 (q, J = 6.7 Hz, 2H); MS m / z [M+H] + 722.47.

[1238] [Production Example 4-228''] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)butanamido)benzoate trifluoroacetic acid

[1239] TIFF2025503429000504.tif35170

[1240] Step 1: 2,5-dioxopyrrolidin-1-yl 4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzoate (Intermediate 4-228''a)

[1241] TIFF2025503429000505.tif35170

[1242] Intermediate 4-228''a was synthesized from Alisertib in the same manner as in Step 4 of Preparation Example 4-177'.

[1243] Step 2: 4-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)butanoic acid (Intermediate 4-228''b)

[1244] TIFF2025503429000506.tif35170

[1245] Intermediate 4-228''b was synthesized from intermediate 4-228''a and 4-aminobutanoic acid in the same manner as in Step 5 of Preparation Example 4-177'.

[1246] Step 3: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)butanamido)benzoate trifluoroacetic acid

[1247] Example 4-228'' was synthesized from Intermediate 4-228''b and Intermediate 3-25a in the same manner as in Steps 2 and 3 of Preparation Example 5-155'.

[1248] 1H NMR (500 MHz, DMSO-d6) δ 10.40 (s, 1H), 10.19 (s, 1H), 8.72 (s, 1H), 8.29 (d, J = 8.6 Hz, 5H), 8.16 - 8.13 (m, 2H), 7.94 (s, 1H), 7.89 (d, J = 8.6 Hz, 1H), 7.84 - 7.79 (m, 2H), 7.52 (dd, J = 8.7, 2.2 Hz, 1H), 7.44 - 7.39 (m, 2H), 7.22 (s, 1H), 4.07 (d, J = 5.7 Hz, 2H), 3.91 (s, 3H), 3.81 (s, MS m / z [M+H] + 760.51.

[1249] [Production Example 5-8'']N-(4-((4-(3-aminoprop-1-yn-1-yl)-3-(hydroxymethyl)phenyl)amino)-4-oxobutyl)-4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide trifluoroacetic acid

[1250] TIFF2025503429000507.tif39170

[1251] Example 5-8'' was synthesized from Alisertib in the same manner as in Example 5-8', and further purified by prep HPLC.

[1252] 1H NMR (500 MHz, DMSO-d6) δ 10.19 (s, 1H), 10.16 (s, 1H), 8.72 (s, 1H), 8.30 (d, J = 8.5 Hz, 1H), 8.25 (s, 4H), 8.14 (t, J = 5.8 Hz, 1H), 7.95 (s, 1H), 7.84 - 7.77 (m, 2H), 7.76 (d, J = 2.2 Hz, 1H), 7.62 (dd, J = 8.4, 2.2 Hz, 1H), 7.41 (ddd, J = 8.6, 6.9, 1.7 Hz, 3H), 7.33 (d, J = 8.4 Hz, 1H), 7.21 MS m / z [M+H] + 762.54.

[1253] [Production Example 5-18'']methyl 2-(3-aminoprop-1-yn-1-yl)-4-((3-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)propyl)carbamoyl)benzoate hydrochloride

[1254] TIFF2025503429000508.tif43170

[1255] Example 5-18'' was synthesized from Alisertib in the same manner as in Preparation Example 5-18'.

[1256] 1H NMR (500 MHz, DMSO-d6) δ 10.22 (s, 1H), 8.89 (t, J = 5.8 Hz, 1H), 8.73 (s, 1H), 8.45 (s, 3H), 8.34 - 8.27 (m, 2H), 8.09 (t, J = 1.2 Hz, 1H), 7.99 (d, J = 1.2 Hz, 2H), 7.96 (s, 1H), 7.88 - 7.79 (m, 3H), 7.48 - 7.38 (m, 3H), 7.25 (s, 1H), 4.06 (q, J = 5.6 Hz, 2H), 3.94 (s, 6H), 3.89 (s, 3H), 3.87 (s, 2H), 3.34 (q, J = 7.4 Hz, 4H), 1.75 (t, J = 6.7 Hz, 2H); MS m / z [M+H] + 790.41.

[1257] [Production Example 5-35'']methyl 2-(3-aminoprop-1-yn-1-yl)-4-((4-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)cyclohexyl)amino)benzoate trifluoroacetic acid

[1258] TIFF2025503429000509.tif34170

[1259] Example 5-35'' was synthesized from Alisertib in the same manner as in Preparation Example 5-35'.

[1260] 1H NMR (500 MHz, DMSO-d6) δ 10.19 (s, 1H), 8.69 (d, J = 3.5 Hz, 1H), 8.29 - 8.21 (m, 6H), 7.99 (d, J = 7.1 Hz, 2H), 7.81 (d, J = 8.6 Hz, 1H), 7.78 (dd, J = 8.5, 2.2 Hz, 2H), 7.68 (dd, J = 8.9, 1.5 Hz, 1H), 7.42 - 7.35 (m, 3H), 6.69 (s, 2H), 3.99 (q, J = 5.7 Hz, 2H), 3.95 (s, 3H), 3.89 (s, 1H), 3.71 (s, 1H), 3.44 (s, 1H), 2.59 (dp, J = 5.5, 1.8 Hz, 2H), 1.94 (d, J = 11.0 Hz, 2H), 1.71 (t, J = 13.3 Hz, 4H), 1.49 (d, J = 9.0 Hz, 2H); MS m / z [M+H] + 802.50.

[1261] [Picture 5-81'']N-(4-((3-(3-aminoprop-1-yn-1-yl)-4-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)amino)-4-oxobutyl) -4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide trifluoroacetic acid

[1262] TIFF2025503429000510.tif41170

[1263] Alisertib is available as a 5-81'

[1264] 1H NMR (500 MHz, DMSO-d6) δ 10.34 (s, 1H), 10.19 (s, 1H), 9.28 (s, 2H), 8.72 (s, 1H), 8.29 (d, J = 8.6 Hz, 1H), 8.26 - 8.17 (m, 4H), 8.14 (t, J = 5.8 Hz, 1H), 7.94 (s, 1H), 7.85 - 7.76 (m, 2H), 7.66 - 7.60 (m, 2H), 7.45 - 7.37 (m, 3H), 7.22 (s, 1H), 3.98 - 3.93 (m, 2H), 3.92 (s, 6H), 3.37 (d, J = 6.4 Hz, 4H), 2.42 (d, J = 7.4 Hz, 2H), 1.87 (t, J = 7.1 Hz, 2H); + 878.42.

[1265] [Production Example 5-104'']N-(6-((3-(3-aminoprop-1-yn-1-yl)phenyl)amino)-6-oxohexyl)-4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamide trifluoroacetic acid

[1266] TIFF2025503429000511.tif39170

[1267] Example 5-104'' was synthesized from Intermediate 4-165''b and Intermediate 1-3a in the same manner as in Steps 3 and 4 of Preparation Example 4-165''.

[1268] 1H NMR (500 MHz, DMSO-d6) δ 10.15 (s, 1H), 10.01 (s, 1H), 8.68 (s, 1H), 8.26 (d, J = 8.5 Hz, 2H), 8.23 ​​(s, 3H), 8.01 (t, J = 5.8 Hz, 1H), 7.93 (t, J = 1.9 Hz, 1H), 7.91 (s, 1H), 7.78 (dd, J = 8.5, 2.2 Hz, 1H), 7.75 (d, J = 8.6 Hz, 1H), 7.43 - 7.39 (m, 2H), 7.37 (dt, J = 8.7, 1.8 Hz, 2H), 7.29 (t, J = 7.9 Hz, 1H), 7.07 (dt, J = 7.6, 1.4 Hz, 1H), 3.97 (d, J = 5.7 Hz, 2H), 3.86 (s, 3H), 2.29 (t, J = 7.4 Hz, 4H), 1.60 (d, J = 7.5 Hz, 2H), 1.50 (d, J = 7.5 Hz, 2H), 1.32 (d, J = 7.4 Hz, 2H); MS m / z [M+H] + 760.44.

[1269] [Production Example 5-150'']methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)-N-methylbutanamido)benzoate hydrochloride

[1270] TIFF2025503429000512.tif35170

[1271] Alisertib is produced by the same method as in Production Example 5-150' and is synthesized in Example 5-150'.

[1272] 1H NMR (500 MHz, DMSO-d6) δ 10.15 (s, 1H), 8.69 (s, 1H), 8.29 - 8.23 ​​(m, 5H), 7.99 (s, 1H), 7.89 (s, 1H), 7.87 (d, J = 8.4 Hz, 1H), 7.79 (dd, J = 8.5, 2.2 Hz, 1H), 7.71 (d, J = 8.6 Hz, 1H), 7.54 (d, J = 2.3 Hz, 1H), 7.47 (dd, J = 8.5, 2.2 Hz, 1H), 7.42 - 7.34 (m, 3H), 7.19 (s, 1H), 4.02 (q, J = 5.6 MS m / z [M+H] + 804.39

[1273] [Manufacturing example III''] A TBM-UBM compound in which a compound (alisertib) exhibiting activity as an Aurora A kinase inhibitor is linked to a compound corresponding to formula III of the present invention was synthesized as follows.

[1274] [Production Example 5-23'']methyl 2-(3-aminoprop-1-yn-1-yl)-4-(1-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzoyl)piperidine-4-carboxamido)benzoate hydrochloride

[1275] TIFF2025503429000513.tif37170

[1276] Example 5-23'' was synthesized from Alisertib in the same manner as in Preparation Example 5-23'.

[1277] 1 H NMR (500 MHz, DMSO-d6) δ 10.46 (s, 1H), 10.04 (s, 1H), 8.66 (s, 1H), 8.33 (s, 3H), 8.26 (d, J = 8.5 Hz, 2H), 8.12 (d, J = 16.3 Hz, 1H), 7.86 (d, J = 8.7 Hz, 2H), 7.78 (dd, J = 8.5, 2.2 Hz, 1H), 7.54 - 7.52 (m, 1H), 7.39 (td, J = 8.4, 6.9 Hz, 2H), 7.34 (s, 1H), 7.19 (d, J = 10.7 Hz, 1H), 7.13 - 7.02 (m, 1H), 4.51 (s, 1H), 4.01 (t, J = 5.6 Hz, 2H), 3.79 (s, 3H), 3.77 (s, 3H), 2.76 (t, J = 12.4 Hz, 2H), 2.59 (dp, J = 5.5, 1.9 Hz, 2H), 1.87 (s, 2H), 1.71 (s, 2H); MS m / z [M+H] + 816.48.

[1278] [Production Example 5-28'']methyl 2-(3-aminoprop-1-yn-1-yl)-4-(3-(1-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzoyl)piperidine-4-carboxamido)prop-1-yn-1-yl)benzoate trifluoroacetic acid

[1279] TIFF2025503429000514.tif37170

[1280] Example 5-28'' was synthesized from Alisertib in the same manner as in Preparation Example 5-28'.

[1281] 1H NMR (500 MHz, DMSO-d6) δ 10.02 (s, 1H), 8.65 (s, 1H), 8.40 (s, 1H), 8.26 (s, 3H), 8.24 (s, 2H), 7.86 (d, J = 8.1 Hz, 1H), 7.84 (s, 1H), 7.79 - 7.75 (m, 1H), 7.55 (s, 1H), 7.52 (d, J = 8.2 Hz, 1H), 7.41 - 7.35 (m, 1H), 7.33 (s, 1H), 7.17 (d, J = 7.5 Hz, 1H), 7.16 - 7.07 (m, 1H), 7.12 - 7.00 (m, 1H), 4.46 (s, 1H), 4.11 (s, 2H), 4.01 (q, J = 5.6 Hz, 2H), 3.82 (s, 3H), 3.76 (s, 6H), 2.93 (d, J = 47.2 Hz, 2H), 2.72 (t, J = MS m / z [M+H] + 854.44.

[1282] [Production Example 5-29'']methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-((1-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzoyl)piperidin-4-yl)methyl)piperazin-1-yl)benzoate trifluoroacetic acid

[1283] TIFF2025503429000515.tif37170

[1284] Example 5-29'' was synthesized from Alisertib and intermediate 5-29d in the same manner as in steps 2 and 3 of Preparation Example 5-155'.

[1285] 11H NMR (500 MHz, DMSO-d6) δ 10.04 (s, 1H), 9.60 (s, 1H), 8.66 (s, 1H), 8.30 (s, 3H), 8.25 (d, J = 8.5 Hz, 1H), 7.81 (d, J = 8.9 Hz, 2H), 7.77 (d, J = 8.8 Hz, 1H), 7.39 (td, J = 8.4, 6.8 Hz, 1H), 7.33 (s, 1H), 7.17 (d, J = 9.3 Hz, 1H), 7.11 (d, J = 2.6 Hz, 1H), 7.09 (d, J = 2.6 Hz, 1H), 7.06 (d, J = 2.7 Hz, 2H), 4.48 (d, J = 12.7 Hz, 1H), 4.00 (d, J = 5.6 Hz, 3H), 3.77 (s, 6H), 3.40 (d, J = 13.0 Hz, 2H), 3.18 (s, 3H), 3.08 (s, 5H), 2.91 (s, 1H), 2.73 (t, J = 12.4 Hz, 1H), 2.08 (s, 1H), 1.83 (d, J = 11.0 Hz, 1H), 1.64 (d, J = 24.6 Hz, 1H), 1.16 (d, J = 37.6 Hz, 3H), 1.00 (d, J = 6.1 Hz, 1H); MS m / z [M+H] + 871.53.

[1286] [Production Example 5-40''] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(6-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzoyl)-2,6-diazaspiro[3.3]heptan-2-yl)benzoate trifluoroacetic acid

[1287] TIFF2025503429000516.tif37170

[1288] Example 5-40'' was synthesized from Alisertib in the same manner as in Preparation Example 5-40'.

[1289] 1 H NMR (500 MHz, DMSO-d6) δ 10.08 (s, 1H), 8.67 (s, 1H), 8.30 - 8.16 (m, 5H), 7.87 (s, 1H), 7.80 - 7.73 (m, 3H), 7.42 - 7.35 (m, 1H), 7.32 (dd, J = 8.5, 1.8 Hz, 1H), 7.25 (d, J = 8.4 Hz, 1H), 7.18 (s, 1H), 6.45 (dd, J = 8.8, 2.5 Hz, 1H), 6.41 (d, J = 2.4 Hz, 1H), 4.17 (s, 2H), 4.12 (s, 2H), 4.09 (d, J = 8.7 Hz, 2H), 4.05 (s, 2H), 3.99 (q, J = 5.6 Hz, 2H), 3.81 (s, 3H), 3.73 (s, 3H); MS m / z [M+H] + 786.41.

[1290] [Manufacturing example IV''] A TBM-UBM compound in which a compound (alisertib) exhibiting activity as an Aurora A kinase inhibitor is linked to a compound corresponding to Formula IV of the present invention was synthesized as follows.

[1291] [Production Example 5-32'']methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(3-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)propanoyl)piperazin-1-yl)benzoate trifluoroacetic acid

[1292] TIFF2025503429000517.tif43170

[1293] Step 1: 3-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)propanoic acid (intermediate 5-32''a)

[1294] TIFF2025503429000518.tif40170

[1295] Intermediate 5-32''a was synthesized from Alisertib in the same manner as in Step 1 of Preparation Example 4-124'.

[1296] Step 2: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(4-(3-(4-((9-chloro-7-(2-fluoro-6-methoxyphenyl)-5H-benzo[c]pyrimido[4,5-e]azepin-2-yl)amino)-2-methoxybenzamido)propanoyl)piperazin-1-yl)benzoate trifluoroacetic acid

[1297] Example 5-32'' was synthesized from Intermediate 5-32''a and Intermediate 5-7b in the same manner as in Production Example 1-1', and then further purified by prep HPLC.

[1298] 1H NMR (500 MHz, DMSO-d6) δ 10.18 (s, 1H), 8.69 (s, 1H), 8.39 (t, J = 5.9 Hz, 1H), 8.27 (s, 1H), 8.25 (s, 3H), 7.94 (s, 1H), 7.83 (d, J = 8.6 Hz, 1H), 7.80 (d, J = 2.2 Hz, 1H), 7.77 (d, J = 8.9 Hz, 2H), 7.41 - 7.38 (m, 1H), 7.37 (dd, J = 8.5, 1.8 Hz, 2H), 7.19 (s, 1H), 7.01 (dd, J = 9.1, 2.7 Hz, 1H), 6.95 (d, J = 2.7 Hz, 1H), 3.99 (q, J = 5.6 Hz, 2H), 3.88 (s, 3H), 3.74 (s, 6H), 3.50 (d, J = 6.3 Hz, 8H), 3.34 (s, 2H), 3.31 (d, J = 5.5 Hz, 2H), 2.62 - 2.59 (m, 2H); MS m / z [M+H] + 845.55.

[1299] [Manufacturing example I'''] A TBM-UBM compound in which a compound (N-cyclopropyl-4'-(4-(2-hydroxyethoxy)benzoyl)-6-methyl-[1,1'-biphenyl]-3-carboxamide) exhibiting p38 MAPK inhibitor activity is linked to a compound corresponding to Formula I of the present invention was synthesized as follows.

[1300] [Production Example 4-166''']2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl(3-(5-(3-aminoprop-1-yn-1-yl)furan-2-carboxamido)propyl)carbamate trifluoroacetic acid

[1301] TIFF2025503429000519.tif42170

[1302] Step 1: tert-butyl(3-(5-((3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propyl)carbamoyl)furan-2-yl)prop-2-yn-1-yl)carbamate (Intermediate 4-166a)

[1303] TIFF2025503429000520.tif12170

[1304] Intermediate 4-166a was synthesized in the same manner as in Step 5 of Production Example 4-1′ using (9H-fluoren-9-yl)methyl(3-aminopropyl)carbamate as a starting material.

[1305] Step 2: tert-butyl(3-(5-((3-aminopropyl)carbamoyl)furan-2-yl)prop-2-yn-1-yl)carbamate (Intermediate 4-166b)

[1306] TIFF2025503429000521.tif12170

[1307] Starting from Intermediate 4-166a, Intermediate 4-166b was synthesized in the same manner as in Step 4 of Preparation Example 5-29.

[1308] Step 3: (4-bromophenyl)(4-(2-hydroxyethoxy)phenyl)methanone (Intermediate 4-166''a)

[1309] TIFF2025503429000522.tif33170

[1310] (4-Bromophenyl)(4-hydroxyphenyl)methanone (1.0 mmol, 1.0 eq), 2-bromoethan-1-ol (1.5 eq), and K2CO3 (2.0 eq) were stirred in acetone at 85 °C for 16 hours. The reaction mixture was purified via flash column chromatography to give intermediate 4-166'''a.

[1311] Step 4: N-cyclopropyl-4'-(4-(2-hydroxyethoxy)benzoyl)-6-methyl-[1,1'-biphenyl]-3-carboxamide (Intermediate 4-166''b)

[1312] TIFF2025503429000523.tif38170

[1313] Intermediate 4-166'''a (0.7 mmol, 1.0 eq), N-cyclopropyl-4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (1.5 eq), Pd(PPh3)4 (0.1 eq), K2CO3 (2.0 eq), and 3.0 mL of DMF were mixed and the reaction vessel was purged with Ar gas. The mixture was then heated at 90 °C for 16 h. The reaction mixture was extracted with EtOAc and H2O, and the organic layer was dried over Na2SO4. The mixture was filtered under reduced pressure, distilled, and purified via flash column chromatography to give intermediate 4-166'''b.

[1314] Step 5: 2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl(4-nitrophenyl)carbonate (intermediate 4-166'''c)

[1315] TIFF2025503429000524.tif43170

[1316] Intermediate 4-166'''b (0.7 mmol, 1.0 eq), 4-nitrophenyl chloroformate (1.1 eq), and TEA (2.0 eq) were stirred in THF at 0 °C for 2 hours. The reaction mixture was purified via flash column chromatography to give intermediate 4-166'''c.

[1317] Step 6: tert-butyl(3-(5-((3-(((2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethoxy)carbonyl)amino)propyl)carbamoyl)furan-2-yl)prop-2-yn-1-yl)carbamate (intermediate 4-166'''d)

[1318] TIFF2025503429000525.tif42170

[1319] Intermediate 4-166'''c (0.5 mmol, 1.0 eq), Intermediate 4-166b (1.0 eq), and DIPEA (3.0 eq) were stirred in DMF at room temperature for 90 minutes. The reaction mixture was purified via flash column chromatography to give Intermediate 4-166'''d.

[1320] Step 7: 2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl(3-(5-(3-aminoprop-1-yn-1-yl)furan-2-carboxamido)propyl)carbamate trifluoroacetic acid

[1321] Starting from Intermediate 4-166'''d, Example 4-166''' was synthesized in the same manner as in Step 3 of Preparation 5-155'.

[1322] 1H NMR (500 MHz, DMSO-d6) δ 8.50 (t, J = 5.8 Hz, 1H), 8.46 (d, J = 4.2 Hz, 1H), 8.35 (s, 2H), 7.79 (dd, J = 8.5, 6.4 Hz, 5H), 7.74 (d, J = 1.9 Hz, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.42 (d, J = 8.0 Hz, 1H), 7.34 (t, J = 5.7 Hz, 1H), 7.17 - 7.12 (m, 3H), 6.99 (d, J = 3.7 Hz, 1H), 4.33 - 4.26 (m, 4H), 4.10 (s, 2H), 3.21 (q, J = 6.7 Hz, 2H), 3.04 - 2.98 (m, 2H), 2.85 (td, J = 7.3, 3.8 Hz, 1H), 2.31 (s, 3H), 1.62 (p, J = 7.0 Hz, 2H), 0.68 (td, J = 7.1, 4.7 Hz, 2H), 0.59 - 0.54 (m, 2H); MS m / z [M+H] + 663.53.

[1323] [Production Example 5-106’’’] methyl 2-(3-aminoprop-1-yn-1-yl)-4-(5-(2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethoxy)-5-oxopentanamido)benzoate hydrochloride

[1324] TIFF2025503429000526.tif23170

[1325] Step 1: 5-((3-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)-4-(methoxycarbonyl)phenyl)amino)-5-oxopentanoic acid (Intermediate 5-106a)

[1326] TIFF2025503429000527.tif19170

[1327] Starting from Intermediate 3-25a, Intermediate 5-106a was synthesized in the same manner as in Step 3 of Preparation Example 5-106′.

[1328] Step 2: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(5-(2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethoxy)-5-oxopentanamido)benzoate hydrochloride

[1329] Example 5-106''' was synthesized from Intermediate 5-106a and Intermediate 4-166'''c in the same manner as in Step 2 of Preparation Example 5-77'.

[1330] 1H NMR (500 MHz, DMSO-d6) δ 10.36 (s, 1H), 8.42 (d, J = 4.2 Hz, 1H), 8.08 (d, J = 2.2 Hz, 1H), 7.84 (d, J = 8.7 Hz, 1H), 7.78 - 7.72 (m, 5H), 7.70 (d, J = 2.0 Hz, 1H), 7.55 - 7.50 (m, 2H), 7.48 (dd, J = 8.7, 2.2 Hz, 1H), 7.38 (d, J = 8.0 Hz, 1H), 7.12 - 7.06 (m, 2H), 4.39 - 4.35 (m, 2H), 4.31 - 4.26 (m, 2H), 3.99 (s, 2H), 3.78 (s, 3H), 2.82 (tt, J = 7.3, 4.0 Hz, 1H), 2.41 - 2.35 (m, 4H), 2.27 (s, 3H), 1.81 (p, J = 7.5 Hz, 2H), 0.67 - 0.62 (m, 2H), 0.54 - 0.50 (m, 2H); MS m / z [M+H] + 716.48.

[1331] [Manufacturing example II'''] A TBM-UBM compound in which a compound (N-cyclopropyl-4'-(4-(2-hydroxyethoxy)benzoyl)-6-methyl-[1,1'-biphenyl]-3-carboxamide) exhibiting p38 MAPK inhibitor activity is linked to a compound corresponding to Formula II of the present invention was synthesized as follows.

[1332] [Production Example 1-1''']2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl(4-(3-aminoprop-1-yn-1-yl)phenyl)carbamate hydrochloride

[1333] TIFF2025503429000528.tif35170

[1334] Step 1: 2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl(4-(3-((tert-butoxycarbonyl)amino)prop-1-yn-1-yl)phenyl)carbamate(intermediate 1-1'''a)

[1335] TIFF2025503429000529.tif39170

[1336] Intermediate 1-1'''a was synthesized in the same manner as in Step 4 of Preparation Example 5-140''' using Intermediate 4-166'''c and Intermediate 1-1a as starting materials.

[1337] Step 2: 2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl(4-(3-aminoprop-1-yn-1-yl)phenyl)carbamate

[1338] Intermediate 1-1''a is produced by the same method as Production Example 1-1 and Example 1-1'' is synthesized.

[1339] 1H NMR (500 MHz, DMSO-d6) δ 8.42 (d, J = 4.3 Hz, 1H), 8.24 (s, 3H), 7.90 (t, J = 5.7 Hz, 1H), 7.86 (d, J = 8.2 Hz, 1H), 7.75 (ddd, J = 8.4, 4.5, 2.4 Hz, 5H), 7.71 (d, J = 2.0 Hz, 1H), 7.57 (d, J = 1.7 Hz, 1H), 7.52 (dd, J = 6.8, 4.8 Hz, 3H), 7.38 (d, J = 8.0 Hz, 1H), 7.13 - 7.08 (m, 2H), 4.33 (d, J = 4.7 Hz, 2H), 4.27 (s, 2H), 4.06 (d, J = 5.7 Hz, 2H), 4.00 (s, 2H), 3.83 (d, J = 3.7 Hz, 3H), 2.81 (td, J = 7.3, 3.8 Hz, 1H), 2.28 (s, 3H), 0.67 - 0.63 (m, 2H), 0.52 (dt, J = 6.9, 4.4 Hz, 2H); MS m / z [M+H] + 684.51

[1340] [Manufacturing Example 3-49''']2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl(4-(3-amino-4-hydroxybut-1-yn-1-yl)phenyl)carbamate hydrochloride

[1341] TIFF2025503429000530.tif49170

[1342] Step 1: 2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl(4-(3-amino-4-hydroxybut-1-yn-1-yl)phenyl)carbamate hydrochloride

[1343] Example 3-49''' was synthesized using Intermediate 3-49b as a starting material in the same manner as in Steps 1 and 2 of Preparation Example 1-1'''.

[1344] 1H NMR (500 MHz, DMSO-d6) δ 10.13 (s, 1H), 8.46 (d, J = 4.3 Hz, 1H), 8.40 (s, 2H), 7.80 (dd, J = 12.2, 8.5 Hz, 5H), 7.74 (d, J = 1.9 Hz, 1H), 7.55 (dd, J = 16.1, 8.6 Hz, 4H), 7.41 (dd, J = 11.5, 8.4 Hz, 3H), 7.16 (d, J = 8.9 Hz, 2H), 5.74 (t, J = 5.3 Hz, 1H), 4.52 - 4.45 (m, 2H), 4.41 - 4.35 (m, 2H), 4.31 (s, 1H), 3.79 - 3.73 (m, 1H), 3.67 - 3.60 (m, 1H), 2.88 - 2.82 (m, 1H), 2.31 (s, 3H), 0.68 (dd, J = 7.1, 2.4 Hz, 2H), 0.56 (dd, J = 4.1, 2.4 Hz, 2H); MS m / z [M+H] + 618.47.

[1345] [Production Example 5-116''']methyl 2-(3-aminoprop-1-yn-1-yl)-4-(3-(((2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethoxy)carbonyl)amino)prop-1-yn-1-yl)benzoate hydrochloride

[1346] TIFF2025503429000531.tif28170

[1347] Step 1: methyl 2-(3-aminoprop-1-yn-1-yl)-4-(3-(((2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethoxy)carbonyl)amino)prop-1-yn-1-yl)benzoate hydrochloride

[1348] Example 5-116''' was synthesized using Intermediate 5-116c as a starting material in the same manner as in Steps 1 and 2 of Preparation Example 1-1'''.

[1349] 1H NMR (500 MHz, DMSO-d6) δ 8.42 (d, J = 4.3 Hz, 1H), 8.24 (s, 3H), 7.90 (t, J = 5.7 Hz, 1H), 7.86 (d, J = 8.2 Hz, 1H), 7.75 (ddd, J = 8.4, 4.5, 2.4 Hz, 5H), 7.71 (d, J = 2.0 Hz, 1H), 7.57 (d, J = 1.7 Hz, 1H), 7.52 (dd, J = 6.8, 4.8 Hz, 3H), 7.38 (d, J = 8.0 Hz, 1H), 7.13 - 7.08 (m, 2H), 4.33 (d, J = 4.7 Hz, 2H), 4.27 (s, 2H), 4.06 (d, J = 5.7 Hz, 2H), 4.00 (s, 2H), 3.83 (d, J = 3.7 Hz, 3H), 2.81 (td, J = 7.3, 3.8 Hz, 1H), 2.28 (s, 3H), 0.67 - 0.63 (m, 2H), 0.52 (dt, J = 6.9, 4.4 Hz, 2H); MS m / z [M+H] + 684.51

[1350] [Manufacturing example III'''] A TBM-UBM compound in which a compound (N-cyclopropyl-4'-(4-(2-hydroxyethoxy)benzoyl)-6-methyl-[1,1'-biphenyl]-3-carboxamide) exhibiting p38 MAPK inhibitor activity is linked to a compound corresponding to Formula III of the present invention was synthesized as follows.

[1351] [Production Example 5-39''']2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl 7-(3-(3-aminoprop-1-yn-1-yl)-4-(methoxycarbonyl)benzoyl)-2,7-diazaspiro[3.5]nonane-2-carboxylate trifluoroacetic acid

[1352] TIFF2025503429000532.tif34170

[1353] Starting from intermediate 5-39c, Example 5-39''' was synthesized in the same manner as in Steps 4 and 5 of Preparation Example 5-140'''.

[1354] 1H NMR (500 MHz, DMSO-d6) δ 8.46 (d, J = 4.3 Hz, 1H), 8.31 (s, 2H), 7.82 - 7.76 (m, 5H), 7.74 (d, J = 1.9 Hz, 1H), 7.61 - 7.55 (m, 3H), 7.52 - 7.46 (m, 2H), 7.42 (d, J = 8.3 Hz, 1H), 7.14 (d, J = 8.9 Hz, 2H), 4.33 (s, 2H), 4.29 (s, 2H), 3.99 (s, 2H), 3.66 (s, 7H), 3.17 (d, J = 8.5 Hz, 2H), 2.85 MS m / z [M+H] + 783.51.

[1355] [Production Example 5-140''']2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl 4-(3-(3-aminoprop-1-yn-1-yl)-4-(methoxycarbonyl)benzoyl)piperazine-1-carboxylate trifluoroacetic acid

[1356] TIFF2025503429000533.tif52170

[1357] Step 1: tert-butyl 4-(3-iodo-4-(methoxycarbonyl)benzoyl)piperazine-1-carboxylate (Intermediate 5-140a)

[1358] TIFF2025503429000534.tif20170

[1359] Intermediate 5-140a was synthesized in the same manner as in Step 5 of Production Example 4-1′ using tert-butyl piperazine-1-carboxylate and 3-iodo-4-(methoxycarbonyl)benzoic acid as starting materials.

[1360] Step 2: tert-butyl 4-(3-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(methoxycarbonyl)benzoyl)piperazine-1-carboxylate (Intermediate 5-140b)

[1361] TIFF2025503429000535.tif38170

[1362] Intermediate 5-140b was synthesized from Intermediate 5-140a in the same manner as in Step 1 of Preparation Example 5-35'.

[1363] Step 3: Methyl 2-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(piperazine-1-carbonyl)benzoate trifluoroacetic acid (Intermediate 5-140c)

[1364] TIFF2025503429000536.tif38170

[1365] Intermediate 5-140c was synthesized from Intermediate 5-140b in the same manner as in Step 4 of Production Example 4-1′.

[1366] Step 4: 2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl 4-(3-(3-(1,3-dioxoisoindolin-2-yl)prop-1-yn-1-yl)-4-(methoxycarbonyl)benzoyl)piperazine-1-carboxylate (intermediate 5-140'''a)

[1367] TIFF2025503429000537.tif59170

[1368] Intermediate 5-140'''a was synthesized from intermediates 5-140c and 4-166'''c in the same manner as in Step 5 of Preparation Example 4-1'.

[1369] Step 5: 2-(4-(5'-(cyclopropylcarbamoyl)-2'-methyl-[1,1'-biphenyl]-4-carbonyl)phenoxy)ethyl 4-(3-(3-aminoprop-1-yn-1-yl)-4-(methoxycarbonyl)benzoyl)piperazine-1-carboxylate trifluoroacetic acid

[1370] Example 5-140''' was synthesized from Intermediate 5-140'''a in the same manner as in Step 5 of Preparation 5-85'.

[1371] 1H NMR (500 MHz, DMSO-d6) δ 8.42 (d, J = 4.3 Hz, 1H), 8.26 (s, 3H), 8.17 (d, J = 8.5 Hz, 1H), 8.06 - 8.00 (m, 1H), 7.92 (d, J = 8.1 Hz, 1H), 7.78 - 7.73 (m, 5H), 7.71 (d, J = 2.0 Hz, 1H), 7.57 (d, J = 1.7 Hz, 1H), 7.53 (d, J = 8.2 Hz, 2H), 7.38 (d, J = 8.1 Hz, 1H), 7.13 (dd, J = 8.9, 2.6 Hz, 1H), 4.36 (d, J = 5.0 Hz, 2H), 4.30 (s, 2H), 4.03 (q, J = 5.7 Hz, 2H), 3.84 (s, 3H), 3.34 (s, 4H), 3.24 (s, 4H), 2.81 (dt, J = 7.4, 3.7 MS m / z [M+H] + 743.55.

[1372] [Experimental Example] [Experimental Example 1] Confirmation of the degradative effect of BRD4 degraders in the MM.1S cell line To confirm that the compounds (BRD4 degraders) of Preparation Examples I' to IV' reduce the amount of BRD4 protein, an experiment was carried out using the MM.1S cell line, which is a B cell line derived from the blood of a multiple myeloma patient.

[1373] Specifically, MM.1S cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and penicillin / streptomycin (100 U / ml) and then transferred to 6-well plates. After 24 hours, BRD4 degraders were diluted in the culture medium and treated at 10 μM increments. After treatment, the cells were cultured in a CO2 incubator for 24 hours. The degradative activity of each compound against intracellular BRD4 protein was confirmed by protein immunoblotting (Western blotting). After incubation according to the experimental conditions, the cells were harvested and centrifuged at 1,800 rpm for 3 minutes. The precipitate was lysed with RIPA (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS, 1x Protease Inhibitor Cocktail), 5x sample buffer was added, and the cells were heated at 100°C for 15 minutes. The prepared samples were electrophoresed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the proteins in the gel were transferred to a PVDF membrane using a Trans-Blot Turbo. The membrane was blocked with 5% bovine serum albumin (BSA) solution for 2 hours at room temperature. Antibodies to BRD4 and β-actin were then diluted in 5% BSA and incubated with the membrane overnight at 4°C. After washing five times with PBST (0.5% Tween 20), the membrane was incubated with horse radish peroxidase-conjugated (HRP) secondary antibodies diluted in PBST for 2 hours at room temperature and washed five times with PBST. After performing an enzyme-substrate reaction using HRP substrate, the signal was measured using chemi-doc equipment, and the band corresponding to the BRD4 protein detected from the compound of the Preparation Example was quantitatively analyzed using densitometry. The results are shown in Tables 16 to 19 below: + if the decomposition ability of each BRD4 decomposer decreased by less than 40%, ++ if it decreased by 40% to less than 70%, and +++ if it decreased by 70% or more.As shown in the table below, it was confirmed that the compounds of Preparation Examples I' to IV' according to the present invention have excellent resolution.

[1374] [Table 16]

[1375] [Table 17] TIFF2025503429000540.tif110170

[1376] [Table 18]

[1377] [Table 19]

[1378] [Experimental Example 2] Confirmation of the decomposition effect of Aurora A decomposer in the MM.1S cell line

[1379] To confirm that the compounds (Aurora A degraders) of Preparation Examples I" to IV" reduce the amount of Aurora A protein, experiments were carried out using the MM.1S cell line, which is a B cell line derived from the blood of a multiple myeloma patient.

[1380] Specifically, MM.1S cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS), 1% penicillin, and streptomycin (100 U / ml) and then transferred to a 6-well plate. After 24 hours, Aurora A degraders were diluted in RPMI medium and treated at a concentration of 5 μM. The cells were then cultured in a CO2 incubator for 24 hours. Protein immunoblotting (Western blotting) was used to confirm the degradative activity of each compound against intracellular Aurora A protein. After incubation under the specified conditions, the cells were harvested and centrifuged at 1,800 rpm for 3 minutes. The precipitate was lysed in RIPA (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS, 1x Protease Inhibitor Cocktail), 5x sample buffer was added, and the cells were heated at 100°C for 15 minutes. The prepared samples were electrophoresed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the proteins in the gel were transferred to a PVDF membrane using a Trans-Blot Turbo. The membrane was blocked with 5% bovine serum albumin (BSA) solution for 2 hours at room temperature. After that, Aurora A and β-actin antibodies were diluted in 5% BSA solution and incubated with the membrane overnight at 4°C. After washing five times with PBST (0.5% Tween 20), HRP-conjugated (horse radish peroxidase-conjugated) secondary antibodies were diluted in PBST and incubated at room temperature for 2 hours, followed by another five washes with PBST. After performing an enzyme-substrate reaction using HRP substrate, the signal was measured using chemi-doc equipment, and the bands corresponding to the Aurora A protein detected in the compounds of the Preparation Examples were quantitatively analyzed using a densitometer. The results are shown in Tables 20 to 23 below: + if the decomposition ability of each Aurora A decomposer decreased by less than 40%, ++ if it decreased by 40% to less than 70%, and +++ if it decreased by 70% or more.The control compound of Chemical Formula 2 showed 0% resolution, whereas the compounds of Preparation Examples I'' to IV'' according to the present invention were confirmed to have excellent resolution.

[1381] [Table 20]

[1382] [Table 21]

[1383] [Table 22]

[1384] [Table 23]

[1385] [Experimental Example 3] Confirmation of the decomposition effect of p38 degraders in macrophages To confirm that the compounds (p38 degraders) of Preparations I''' to III''' reduce the amount of p-p38 (phospho-p38) protein, experiments were performed using macrophages Raw 264.7. p38 MAPK alpha is activated by upstream regulators such as MKK3 and MKK6 through phosphorylation of threonine dinucleotide 180 (Thr180) and tyrosine 182 (Tyr182). Generally, phosphorylation of these residues is detected using an antibody (3D7 rabbit monoclonal antibody, Cellsignaling) that specifically recognizes p-p38 (phospho-p38) MAPK, which activates p38 MAPK alpha.

[1386] Specifically, RAW 264.7 cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS), 1% penicillin, and streptomycin (100 U / ml) and then transferred to a 6-well plate. After 24 hours, p38 degraders were diluted in DMEM medium and treated at 10 μM. After treatment, the cells were incubated in a CO2 incubator for 24 hours. Protein immunoblotting (Western blotting) was used to confirm the degradative activity of each compound against intracellular p38 and p-p38 proteins. After incubation according to the experimental conditions, the cells were harvested and centrifuged at 1,800 rpm for 3 minutes. The precipitate was lysed with RIPA (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS, 1x Protease Inhibitor Cocktail), 5x sample buffer was added, and the cells were heated at 100°C for 15 minutes. The prepared samples were electrophoresed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the proteins in the gel were transferred to a PVDF membrane using a Trans-Blot Turbo. The membrane was blocked with 5% bovine serum albumin (BSA) solution for 2 hours at room temperature. Antibodies against p-p38, p38, and β-actin were diluted in 5% BSA solution and incubated with the membrane overnight at 4°C. After washing five times with PBST (phosphate-based saline containing 0.5% Tween 20), the membrane was incubated with horse radish peroxidase (HRP)-conjugated secondary antibodies diluted in PBST for 2 hours at room temperature and then washed five times with PBST. After performing an enzyme-substrate reaction using HRP substrate, signals were measured using chemi-doc equipment, and the bands corresponding to p-p38 and p38 protein detected in the compounds of the Preparation Examples were quantitatively analyzed using densitometry. The results are shown in Tables 24 to 26 below: + if the decomposition ability of each p38 decomposer decreased by less than 40%, ++ if it decreased by 40% to less than 70%, and +++ if it decreased by 70% or more.As shown in the table below, it was confirmed that the compounds of Preparation Examples I''' to III''' according to the present invention have excellent resolution.

[1387] [Table 24]

[1388] [Table 25]

[1389] [Table 26]

[1390] [Experimental Example 4] Confirmation of direct ubiquitin binding using pulldown assay To confirm whether the compounds represented by Formulas I to IV of the present invention can bind to ubiquitin and undergo polyubiquitination by various E3 ligase complexes, an in vitro ubiquitination assay was performed using ITCH / AIP4, NEDD4, and cIAP-1 / HUAP-2 proteins (manufactured by R&D Systems) as E3 ligases.

[1391] Specifically, the first lane contained 10X buffer and 100 μg of ubiquitin. Lanes two through six contained 10X buffer, 100 μg of ubiquitin, 5 nM E1, 100 nM E2, 14 nM E3, and 1 mM ATP. The third lane contained biotin at 40 μM as a negative control. The fourth and fifth lanes contained biotin-conjugated compounds (Examples 4-228 and 4-166) at 40 μM. Each sample was incubated in a 37°C incubator for 3 hours. After incubation, 5 μl of each sample was added to 5X sample buffer and heated at 100°C for 10 minutes to determine the input results. The remaining sample solution was added to streptavidin-conjugated agarose beads and incubated at 4°C for 3 hours. After washing five ti...

Claims

1. A compound represented by the following formula I to formula IV, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof: [Chemical formula I] [Chemical formula II] [Chemical formula III] [Chemical formula IV] In the formulas I to IV, each independently represents W １ is -alkylene-, -alkylene-O-, -alkylene-OSO which is unsubstituted or substituted with at least one selected from the group consisting of deuterium, alkyl, C(O)OR', C(O)N(R')(R"), alkylene-C(O)OR', alkylene-C(O)N(R')(R"), alkylene-OR', alkylene-SR', alkylene-N(R')(R"), alkylene-OP(O)(OR')(OR"), alkylene-P(O)(OR')(OR"), and alkylene-OP(O)(R')(R") ２ - and - alkylene-N(R')SO ２ - any one selected from the group consisting of A is any one selected from the group consisting of aryl, heteroaryl, cycloalkyl, heterocycloalkyl, and heterocycloalkenyl; R １ and R ２ are each independently hydrogen, alkyl, alkenyl, alkynyl, halogen, oxo, CN, NO ２ , OR', SR', N(R')(R''), O-(alkylene)-OR', -N(R')-(alkylene)-OR', C(O)R', C(NH)R', C(S)R', C(O)OR', C(O)N(R')(R''), P(O)(R')(R''), P(O)(OR')(OR''), C(NH)N(R')(R''), C(S)OR', C(S)N(R')(R''), C(O)O-(alkylene)-OC(O)R', C(O)O-(A (alkylene)-methacryloyloxy, (alkylene)-OR', (alkylene)-SR', (alkylene)-N(R')(R'', (alkynylene)-N(R')(R'', (alkylene)-P(O)(R')(R'')-, (alkylene)-P(O)(OR')(OR'', (alkylene)-OP(O)(R')(R'', (alkylene)-OP(O)(OR')(OR'', (alkylene)-OP(O)(benzyloxy) ２ , (alkylene)-OP(O)(phenoxy) ２ , (alkylene)-OP(O)(phenoxy)(NH-CH(R')C(O)OR'', (alkylene)-C(O)OR', (alkenylene)-C(O)OR', (alkynylene)-C(O)OR', (alkylene)-C(O)N(R')(R'', (alkenylene)-C(NH)N(R')(R'', (alkynylene)-C(S)N(R')(R'', (alkylene)-C(O)R', (alkylene)-C(NH)R', (alkylene)-C(S)R', aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein R １ and R ２ may each independently be substituted with one or more halogens, and each independently represents １ and R ２ is aryl, heteroaryl, heterocycloalkyl, or heterocycloalkenyl, which may be substituted at one or more positions with alkyl, substituted or unsubstituted with one or more halogens, or N(R')(R''); X １ is a direct bond or -N(R')-, -O-, -S-, -C(O)-, -SO-, SO ２ the X1a group consisting of -, =N-O- and -O-N=, -OC(O)-, -C(O)O-, -N(R')C(O)-, -C(O)N(R')-, -N(R')SO-, -SON(R')- and -N(R')SO ２ - and -SO ２ one selected from the group X1b consisting of N(R')- and the group X1c consisting of -alkylene-, -alkenylene- and -alkynylene-, or two or more of which are linked together; X ２ is a direct bond or -N(R')-, -O-, -S-, -C(O)-, -SO-, SO ２ the X2a group consisting of -, =N-O- and -O-N=, -OC(O)-, -N(R')C(O)-, -N(R')C(O)O- and -N(R')SO ２ The X2b group consists of —O— and —N(R′)C(S)O—, —C(O)O—, —C(O)N(R′)—, —O—SO ２ one selected from the X2c group consisting of -N(R')-, -O-C(S)N(R')- and -O-C(O)N(R')-, and the X2d group consisting of -alkylene-, -alkenylene- and -alkynylene-, or two or more of which are linked together; X ３ is a direct bond or -alkylene-, -O-, -S-, -C(O)-, -N(R')-, -SO-, -SO ２ - and any combination thereof; B is any one selected from the group consisting of aryl, heteroaryl, cycloalkyl, heterocycloalkyl, and heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy, alkoxy, and oxo, or two or more selected from the group consisting of aryl, heteroaryl, cycloalkyl, heterocycloalkyl, and heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy, alkoxy, and oxo, are directly bonded, -alkylene-, -O-, -S-, -C(O)-, -N(R')-, -SO-, -SO ２ - or combinations thereof, W ２ is hydrogen or deuterium, halogen, alkyl, alkylene-C(O)OR', alkylene-C(O)N(R')(R'', alkylene-OR', alkylene-SR', alkylene-N(R')(R'', alkylene-OP(O)(OR')(OR'', alkylene-P(O)(OR')(OR'', alkylene-OP(O)(R')(R'', -N(R')(R'', -OR', -SR', -C(O)R', -SOR', -SO ２ R', =N-OR', -C(O)OR', -C(O)N(R')(R''), -OC(O)OR', -OC(O)N(R')(R''), -N(R')C(O)O(R''), -N(R')SO(R'), -SON(R')(R''), -N(R')SO ２ (R″) and —SO ２ N(R')(R'') is any one selected from the group consisting of alkyl, alkenyl, and alkynyl, each of which may be substituted or unsubstituted with at least one selected from the group consisting of alkyl, alkenyl, and alkynyl; Y １ ' is halogen, -N(R')(R'', -OR', -SR', -C(O)R', -SOR', SO ２ R', =N-OR', -C(O)OR', -C(O)N(R')(R''), -N(R')SO(R'), -SON(R')(R''), -OC(O)OR', -OC(O)N(R')(R''), -N(R')C(O)O(R''), -N(R')SO ２ (R″) and —SO ２ N(R′)(R″) is substituted or unsubstituted with at least one selected from the group consisting of alkyl, alkenyl, alkynyl, (CH ３ CH ２ O)-(CH ２ CH ２ O) ｎ－１ CH ２ - and (CH ３ CH ２ O)-(CH ２ CH ２ O) ｎ－１ CH ２ CH ２ - any one selected from the group consisting of, where n is 1 to 4; Y ２ ' is halogen, -N(R')(R'', -OR', -SR', -C(O)R', -SOR', SO ２ R', =N-OR', -C(O)OR', -C(O)N(R')(R''), -OC(O)OR', -OC(O)N(R')(R''), -N(R')C(O)O(R''), -N(R')SO(R'), -SON(R')(R''), -N(R')SO ２ (R″) and —SO ２ N(R')(R'') is any one selected from the group consisting of alkyl, alkenyl, alkynyl, heterocycloalkyl, and heterocycloalkenyl, each of which may be substituted or unsubstituted with at least one selected from the group consisting of alkyl, alkenyl, alkynyl, heterocycloalkyl, and heterocycloalkenyl; In the above functional groups, R' and R'' are each independently hydrogen or alkyl.

2. In the formulas I to IV, each independently represents The W １ is -alkylene-, -alkylene-O-, -alkylene-OSO, which are unsubstituted or substituted with at least one selected from the group consisting of C(O)OR', C(O)N(R')(R"), alkylene-OR', alkylene-SR', and alkylene-OP(O)(OR')(OR") ２ - and - alkylene-N(R')SO ２ - any one selected from the group consisting of A is any one selected from the group consisting of aryl, 5- to 10-membered heteroaryl containing at least one selected from the group consisting of N, O, and S, 3- to 10-membered cycloalkyl, 3- to 10-membered heterocycloalkyl containing at least one selected from the group consisting of N, O, and S, and 3- to 10-membered heterocycloalkenyl containing at least one selected from the group consisting of N, O, and S; The R １ and R ２ are each independently hydrogen, alkyl, alkenyl, halogen, OR', N(R')(R'', O-(alkylene)-OR', C(O)R', C(O)OR', C(O)N(R')(R'', C(NH)N(R')(R'', C(O)O-(alkylene)-OC(O)R', C(O)O-(alkylene)-methacryloyloxy, (alkylene)-OR', (alkylene)-SR', (alkylene)-N(R')(R'', (alkynylene)-N(R')(R'', (alkylene)-OP(O)(OR')(OR'', (alkylene)-OP(O)(benzyloxy). ２ , (alkylene)-OP(O)(phenoxy) ２ , (alkylene)-OP(O)(phenoxy)(NH-CH(R')C(O)OR'', (alkylene)-C(O)OR', aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl, １ and R ２ may each independently be substituted with one or more halogens, and each independently represents １ and R ２ When is aryl, heteroaryl, heterocycloalkyl, or heterocycloalkenyl, it may be substituted at one or more positions with alkyl, substituted or unsubstituted with one or more halogens, or N(R')(R''); The X １ is a direct bond, any one selected from the X1a group and the X1b group, or any one selected from the X1a group and the X1b group and any one selected from the X1c group are linked in the form of X1a-X1c or X1b-X1c; The X ２ is a direct bond, any one selected from the X2a group, the X2b group, and the X2c group, or two or more of them are linked to each other, or any one or two or more selected from the X2b group, the X2c group, and the X2d group are linked in the form of X2b-X2d-X2b, X2b-X2d-X2c, X2c-X2d-X2b, or X2c-X2d-X2c; The X ３ is a direct bond or any one selected from the group consisting of -C(O)-, -N(R')- and combinations thereof; B is any one selected from the group consisting of aryl, 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy, alkoxy, and oxo, or two or more selected from the group consisting of aryl, 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy and oxo, which are directly linked by -alkylene-, -C(O)-, -N(R')-, or a combination thereof; The W ２ is hydrogen or deuterium, halogen, alkyl, alkylene-C(O)OR', alkylene-C(O)N(R')(R'', alkylene-OR', alkylene-SR', alkylene-N(R')(R'', alkylene-OP(O)(OR')(OR'', alkylene-P(O)(OR')(OR'', alkylene-OP(O)(R')(R'', -N(R')(R'', -OR', -SR', -C(O)R', -SOR', -SO ２ R', =N-OR', -C(O)OR', -C(O)N(R')(R''), -OC(O)OR', -OC(O)N(R')(R''), -N(R')C(O)O(R''), -N(R')SO(R''), -SON(R')(R''), -N(R')SO ２ (R″) and —SO ２ any one selected from the group consisting of alkenyl and alkynyl, each of which may be substituted or unsubstituted with at least one selected from the group consisting of N(R')(R''); The Y １ ' is halogen, -N(R')(R'', -OR', -SR', -C(O)R', -SOR', SO ２ R', =N-OR', -C(O)OR', -C(O)N(R')(R''), -OC(O)OR', -OC(O)N(R')(R''), -N(R')C(O)O(R''), -N(R')SO(R'), -SON(R')(R''), -N(R')SO ２ (R″) and —SO ２ N(R′)(R″) is substituted or unsubstituted with at least one selected from the group consisting of alkyl, alkenyl, alkynyl, (CH ３ CH ２ O)-(CH ２ CH ２ O) ｎ－１ CH ２ - and (CH ３ CH ２ O)-(CH ２ CH ２ O) ｎ－１ CH ２ CH ２ - any one selected from the group consisting of, where n is 1 to 2; The Y ２ ' is halogen, -N(R')(R'', -OR', -SR', -C(O)R', -SOR', SO ２ R', =N-OR', -C(O)OR', -C(O)N(R')(R''), -OC(O)OR', -OC(O)N(R')(R''), -N(R')C(O)O(R''), -N(R')SO(R'), -SON(R')(R''), -N(R')SO ２ (R″) and —SO ２ The compound according to claim 1, which is any one selected from the group consisting of alkyl, alkenyl, alkynyl and heterocycloalkyl, each of which is substituted or unsubstituted with at least one selected from the group consisting of N(R')(R''), or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof.

3. In the formulas I to IV, each independently represents The W １ is -alkylene-, -alkylene-O-, -alkylene-OSO, which is unsubstituted or substituted with at least one selected from the group consisting of alkylene-OR' and alkylene-OP(O)(OR')(OR''). ２ - and - alkylene-N(R')SO ２ - any one selected from the group consisting of A is any one selected from the group consisting of benzene, furan, benzofuran, pyridine, thiophene, imidazole and oxazole; The R １ and R ２ are each independently any one selected from the group consisting of hydrogen, alkyl, OR', C(O)R', C(O)OR', C(O)N(R')(R"), (alkylene)-OR', (alkylene)-SR', (alkylene)-OP(O)(OR')(OR"), heteroaryl, and heterocycloalkyl; １ and R ２ may each independently be substituted with one or more halogens, and each independently represents １ and R ２ is heteroaryl, or heterocycloalkyl, which may be substituted at one or more positions with one or more halogen-substituted or unsubstituted alkyl or N(R')(R''); The X １ is a direct bond, or said X1a group consists of -N(R')- and -O-, and said X1b group consists of -C(O)O-, -N(R')C(O)- and -C(O)N(R')-; The X ２ is a direct bond, or said X2a group consists of N(R')-, -O-, -C(O)- and =N-O-, said X2b group consists of -N(R')C(O)- and said X2c group consists of -C(O)N(R')-; B is any one selected from the group consisting of 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy and oxo, or two or more selected from the group consisting of unsubstituted 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl may be directly linked to each other via -alkylene-, -C(O)-, -N(R')-, or a combination thereof; W ２ ' is hydrogen or alkynyl unsubstituted or substituted with at least one selected from the group consisting of halogen, alkylene-OR', alkylene-OP(O)(OR')(OR"), -N(R')(R"), -OR' and -C(O)OR'; Y １ ' is any one selected from the group consisting of alkyl, alkenyl and alkynyl, each of which is substituted or unsubstituted with at least one selected from the group consisting of halogen, -N(R')(R'', -OR' and -C(O)OR'; Y ２ 3. The compound of claim 2, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof, wherein R′ is any one selected from the group consisting of alkyl and heterocycloalkyl, each of which may be substituted or unsubstituted with at least one selected from the group consisting of halogen, -N(R')(R'', -OR', and -C(O)OR'.

4. The compound according to claim 3, which is at least one selected from the group consisting of the compounds listed in Tables 1 to 4 below, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate, or prodrug thereof. 【Table 1】 【Table 2】 【Table 3】 【Table 4】

5. 2. The compound of claim 1, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof, wherein the compound is a ubiquitin-binding compound.

6. having the chemical structure TBM-UBM, The TBM is a moiety that binds to a target protein or polypeptide; The UBM is a compound having a chemical structure represented by the following formulas V to VIII, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate, or prodrug thereof. [Chemical formula V] [Chemical formula VI] [Chemical formula VII] [Chemical formula VIII] In the formulas V to VIII, each independently represents W １ is -alkylene-, -alkylene-O-, -alkylene-OSO which is unsubstituted or substituted with at least one selected from the group consisting of deuterium, alkyl, C(O)OR', C(O)N(R')(R"), alkylene-C(O)OR', alkylene-C(O)N(R')(R"), alkylene-OR', alkylene-SR', alkylene-N(R')(R"), alkylene-OP(O)(OR')(OR"), alkylene-P(O)(OR')(OR"), and alkylene-OP(O)(R')(R") ２ - and - alkylene-N(R')SO ２ - any one selected from the group consisting of A is any one selected from the group consisting of aryl, heteroaryl, cycloalkyl, heterocycloalkyl, and heterocycloalkenyl; R １ and R ２ are each independently hydrogen, alkyl, alkenyl, alkynyl, halogen, oxo, CN, NO ２ , OR', SR', N(R')(R''), O-(alkylene)-OR', -N(R')-(alkylene)-OR', C(O)R', C(NH)R', C(S)R', C(O)OR', C(O)N(R')(R''), P(O)(R')(R''), P(O)(OR')(OR''), C(NH)N(R')(R''), C(S)OR', C(S)N(R')(R''), C(O)O-(alkylene)-OC(O)R', C(O)O-(A (alkylene)-methacryloyloxy, (alkylene)-OR', (alkylene)-SR', (alkylene)-N(R')(R'', (alkynylene)-N(R')(R'', (alkylene)-P(O)(R')(R'')-, (alkylene)-P(O)(OR')(OR'', (alkylene)-OP(O)(R')(R'', (alkylene)-OP(O)(OR')(OR'', (alkylene)-OP(O)(benzyloxy) ２ , (alkylene)-OP(O)(phenoxy) ２ , (alkylene)-OP(O)(phenoxy)(NH-CH(R')C(O)OR'', (alkylene)-C(O)OR', (alkenylene)-C(O)OR', (alkynylene)-C(O)OR', (alkylene)-C(O)N(R')(R'', (alkenylene)-C(NH)N(R')(R'', (alkynylene)-C(S)N(R')(R'', (alkylene)-C(O)R', (alkylene)-C(NH)R', (alkylene)-C(S)R', aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl, wherein R １ and R ２ may each independently be substituted with one or more halogens, and each independently represents １ and R ２ is aryl, heteroaryl, heterocycloalkyl, or heterocycloalkenyl, which may be substituted at one or more positions with alkyl, substituted or unsubstituted with one or more halogens, or N(R')(R''); X １ is a direct bond or -N(R')-, -O-, -S-, -C(O)-, -SO-, SO ２ the X1a group consisting of -, =N-O- and -O-N=, -OC(O)-, -C(O)O-, -N(R')C(O)-, -C(O)N(R')-, -N(R')SO-, -SON(R')- and -N(R')SO ２ - and -SO ２ one selected from the group X1b consisting of N(R')- and the group X1c consisting of -alkylene-, -alkenylene- and -alkynylene-, or two or more of which are linked together; X ２ is a direct bond or -N(R')-, -O-, -S-, -C(O)-, -SO-, SO ２ the X2a group consisting of -, =N-O- and -O-N=, -OC(O)-, -N(R')C(O)-, -N(R')C(O)O- and -N(R')SO ２ The X2b group consists of —O— and —N(R′)C(S)O—, —C(O)O—, —C(O)N(R′)—, —O—SO ２ one selected from the X2c group consisting of -N(R')-, -O-C(S)N(R')- and -O-C(O)N(R')-, and the X2d group consisting of -alkylene-, -alkenylene- and -alkynylene-, or two or more of which are linked together; X ３ is a direct bond or -alkylene-, -O-, -S-, -C(O)-, -N(R')-, -SO-, -SO ２ - and any combination thereof; B is any one selected from the group consisting of aryl, heteroaryl, cycloalkyl, heterocycloalkyl, and heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy, alkoxy, and oxo, or two or more selected from the group consisting of aryl, heteroaryl, cycloalkyl, heterocycloalkyl, and heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy, alkoxy, and oxo, are bonded together by a direct bond, -alkylene-, -O-, -S-, -C(O)-, -N(R')-, -SO-, -SO ２ - or combinations thereof, W ２ is a direct bond or any one selected from the group consisting of -alkylene-, -alkenylene- and -alkynylene-, which are unsubstituted or substituted with at least one selected from the group consisting of deuterium, alkyl, C(O)OR', C(O)N(R')(R"), alkylene-C(O)OR', alkylene-C(O)N(R')(R"), alkylene-OR', alkylene-SR', alkylene-N(R')(R"), alkylene-OP(O)(OR')(OR"), alkylene-P(O)(OR')(OR") and alkylene-OP(O)(R')(R"); Y １ represents a direct bond, alkylene, alkenylene, alkynylene, -(CH ２ CH ２ O) ｎ CH ２ - and - (CH ２ CH ２ O) ｎ CH ２ CH ２ - any one selected from the group consisting of, where n is 1 to 4; Y ２ is any one selected from the group consisting of a direct bond, alkylene, alkenylene, alkynylene, heterocycloalkylene, and heterocycloalkenylene; The TBM and the UBM are linked by a covalent bond, and the covalent bond site is a direct bond or is selected from the group consisting of -N(R')-, -O-, -C(R')(R'')-O-, -S-, -C(O)-, -SO-, SO ２ -, =N-O-, -O-N=, -OC(O)-, -C(O)O-, -N(R')C(O)-, -C(O)N(R')-, -OC(O)O-, -OC(O)N(R')-, -N(R')C(O)O-, -N(R')SO-, -SON(R')-, -N(R')SO ２ - and -SO ２ TBM and UBM may each be modified to form one selected from the group consisting of N(R′)—; In the above functional groups, R' and R'' are each independently hydrogen or alkyl.

7. In the formulas V to VIII, each independently represents The W １ is -alkylene-, -alkylene-O-, -alkylene-OSO, which are unsubstituted or substituted with at least one selected from the group consisting of C(O)OR', C(O)N(R')(R"), alkylene-OR', alkylene-SR', and alkylene-OP(O)(OR')(OR") ２ - and - alkylene-N(R')SO ２ - any one selected from the group consisting of A is any one selected from the group consisting of aryl, 5- to 10-membered heteroaryl containing at least one selected from the group consisting of N, O, and S, 3- to 10-membered cycloalkyl, 3- to 10-membered heterocycloalkyl containing at least one selected from the group consisting of N, O, and S, and 3- to 10-membered heterocycloalkenyl containing at least one selected from the group consisting of N, O, and S; The R １ and R ２ are each independently hydrogen, alkyl, alkenyl, halogen, OR', N(R')(R'', O-(alkylene)-OR', C(O)R', C(O)OR', C(O)N(R')(R'', C(NH)N(R')(R'', C(O)O-(alkylene)-OC(O)R', C(O)O-(alkylene)-methacryloyloxy, (alkylene)-OR', (alkylene)-SR', (alkylene)-N(R')(R'', (alkynylene)-N(R')(R'', (alkylene)-OP(O)(OR')(OR'', (alkylene)-OP(O)(benzyloxy). ２ , (alkylene)-OP(O)(phenoxy) ２ , (alkylene)-OP(O)(phenoxy)(NH-CH(R')C(O)OR'', (alkylene)-C(O)OR', aryl, heteroaryl, heterocycloalkyl and heterocycloalkenyl, １ and R ２ may each independently be substituted with one or more halogens, and each independently be １ and R ２ is aryl, heteroaryl, heterocycloalkyl, or heterocycloalkenyl, which may be substituted at one or more positions with alkyl or N(R')(R'') substituted or unsubstituted with one or more halogens; The X １ is a direct bond, any one selected from the X1a group and the X1b group, or any one selected from the X1a group and the X1b group and any one selected from the X1c group are linked in the form of X1a-X1c or X1b-X1c; The X ２ is a direct bond, any one selected from the X2a group, the X2b group, and the X2c group, or two or more of them are linked to each other, or any one or two or more selected from the X2b group, the X2c group, and the X2d group are linked in the form of X2b-X2d-X2b, X2b-X2d-X2c, X2c-X2d-X2b, or X2c-X2d-X2c; The X ３ is a direct bond or any one selected from the group consisting of -C(O)-, -N(R')- and combinations thereof; B is any one selected from the group consisting of aryl, 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy, alkoxy, and oxo, or two or more selected from the group consisting of aryl, 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy and oxo, which are directly linked by -alkylene-, -C(O)-, -N(R')-, or a combination thereof; W ２ is a direct bond or any one selected from the group consisting of -alkenylene- and -alkynylene-, which are unsubstituted or substituted with at least one selected from the group consisting of alkylene-OR', alkylene-SR', and alkylene-OP(O)(OR')(OR''); Y １ represents a direct bond, alkylene, alkenylene, alkynylene, -(CH ２ CH ２ O) ｎ CH ２ - and - (CH ２ CH ２ O) ｎ CH ２ CH ２ - any one selected from the group consisting of, where n is 1 to 2; Y ２ is any one selected from the group consisting of a direct bond, alkylene, alkenylene, alkynylene, and heterocycloalkylene; The compound of claim 6 or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof, wherein the covalent bond site at which the TBM and UBM are linked is a direct bond, or the TBM and UBM may each be modified to form any one selected from the group consisting of -N(R')-, -O-, -C(R')(R'')-O-, -OC(O)-, -OC(O)N(R')- and -C(O)N(R')-.

8. In the formulas V to VIII, each independently represents The W １ is -alkylene-, -alkylene-O-, -alkylene-OSO, which is unsubstituted or substituted with at least one selected from the group consisting of alkylene-OR' and alkylene-OP(O)(OR')(OR''). ２ - and - alkylene-N(R')SO ２ - any one selected from the group consisting of A is any one selected from the group consisting of benzene, furan, benzofuran, pyridine, thiophene, imidazole and oxazole; The R １ and R ２ are each independently any one selected from the group consisting of hydrogen, alkyl, OR', C(O)R', C(O)OR', C(O)N(R')(R"), (alkylene)-OR', (alkylene)-SR', (alkylene)-OP(O)(OR')(OR"), heteroaryl, and heterocycloalkyl; １ and R ２ may each independently be substituted with one or more halogens, and each independently represents １ and R ２ is heteroaryl, or heterocycloalkyl, which may be substituted at one or more positions with one or more halogen-substituted or unsubstituted alkyl or N(R')(R''); The X １ is a direct bond, or said X1a group consists of -N(R')- and -O-, and said X1b group consists of -C(O)O-, -N(R')C(O)- and -C(O)N(R')-; The X ２ is a direct bond, or said X2a group consists of N(R')-, -O-, -C(O)- and =N-O-, said X2b group consists of -N(R')C(O)- and said X2c group consists of -C(O)N(R')-; B is any one selected from the group consisting of 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl, which are substituted or unsubstituted with at least one selected from the group consisting of hydroxy and oxo, or two or more selected from the group consisting of unsubstituted 5- to 10-membered heteroaryl, 3- to 10-membered heterocycloalkyl, and 3- to 10-membered heterocycloalkenyl may be directly linked to each other via -alkylene-, -C(O)-, -N(R')-, or a combination thereof; W ２ is a direct bond or -alkynylene- unsubstituted or substituted with at least one selected from the group consisting of alkylene-OR' and alkylene-OP(O)(OR')(OR''); Y １ is any one selected from the group consisting of a direct bond, alkylene, alkenylene, and alkynylene; Y ２ is any one selected from the group consisting of a direct bond, alkylene, and heterocycloalkylene; The compound of claim 7 or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof, wherein the covalent bond site at which the TBM and UBM are linked is a direct bond or each of the TBM and UBM may be modified to form any one selected from the group consisting of -O-, -C(R')(R'')-O-, -OC(O)-, -OC(O)N(R')- and -C(O)N(R')-.

9. 9. The compound of any one of claims 6 to 8, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof, wherein the compound degrades a target protein or polypeptide to which the TBM binds.

10. The target proteins or polypeptides to which the TBM binds include proteins associated with cell death mechanisms, including B7.1, B7, TINFRlm, TNFR2, NADPH oxidase, and Bcl2Bax, proteins associated with the MAP / ERK signaling pathway, including Ras and Raf, proteins associated with the NF-κB signaling pathway, proteins associated with the mTOR signaling pathway, proteins associated with the wnt signaling pathway, proteins associated with the AKT signaling pathway, proteins associated with the JAK-STAT signaling pathway, proteins associated with the NOX complex and its signaling pathway, proteins associated with vascular endothelial growth factors and receptors and their signaling pathways, proteins associated with TLR receptors and their signaling pathways, proteins associated with G protein-linked receptors and their signaling pathways, proteins associated with TCR-MHC complex-associated receptors and their signaling pathways, proteins associated with cell cycle checkpoint proteins and their signaling pathways, proteins including factors regulating gene transcription and translation, proteins associated with gene replication, and and their regulatory factors, proteins including histone protein regulatory factors for gene expression, factors and related proteins regulating epigenetics, proteins that constitute and regulate the cytoskeleton, proteins that regulate intracellular trafficking and proteins that regulate the same, C5a receptor, dopamine receptor, 5HT receptor, chemokine receptor, histamine receptor, CD40 / CD40L, CD23, CD124, CD4, CD5, VCAM, VLA-4 integrin, selectin, ICOS, ICAM1, IL-2 receptor, IL-1 receptor, T NF-alpha receptors and other related cytokine and chemokine receptors, oxytocin receptors, glycine receptors, noradrenaline reuptake receptors, endothelin receptors, estrogen receptors, androgen receptors, adenosine receptors, TrkA receptors for NGF, vitronectin receptors, integrin receptors, neurokinins and receptors, epidermal growth factor receptors, purinergic receptors (P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7), EGF receptors, HMG-CoA reductase, PDE V phosphodiesterase type, PDE IV phosphodiesterase type 4, PDE I, PDE II, PDEIII, nitric oxide (NO) synthase, cyclooxygenase 1, cyclooxygenase 2, 5-lipoxygenase, trypsinase serine protease, thymidylate synthase, purine nucleoside phosphorylase, GAPDH trypanosomal, glycogen phosphorylase, carbonic anhydrase and its analogs, P-glycoprotein and MRP, tyrosine kinase, p56, lck, Cat+ pathway, inosine monophosphate dehydrogenase, interleukin-1 convertase, caspase-1, glycinamide ribonucleoside formyltransferase, poly(ADP-ribose) polymerase, acetylcholinesterase, cyclin-dependent kinase, microsomal transfer protein inhibitors, bile acid transport inhibitors, 5-alpha reductase inhibitors, a angiotensin 11, adenosine receptors, adenosine kinase and AMP deaminase, farnesyl transferase, geranylgeranyl transferase, beta-amyloid, tyrosine kinase Her-21 / neu, telomerase inhibition, cytoplasmic phospholipase A2 and tyrosine kinase, acetyl-CoA carboxylase, adenylosuccinate synthase, protoporphyrinogen oxidase and enolpyruvylshikimate-phosphate synthase, ecdysone 20-monooxygenase, ion pathways of GABA-gated chloride pathways, voltage-sensitive sodium pathway protein, sodium pathway, calcium release pathway protein, chloride pathway protein and polypeptides derived therefrom, HCV 9. The compound according to any one of claims 6 to 8, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof, which is at least one selected from the group consisting of bacterial or viral proteins such as NS3 protease, HCV NS3 RNA helicase, rhinovirus 3C protease, herpes simplex virus-1 (HSV-I) protease, cytomegalovirus (CMV) protease, multidrug resistance (MDR) protein, HIV-1 protease, HIV-1 integrase, influenza, neuraminidase, hepatitis B reverse transcriptase, and polypeptides derived therefrom, proteins such as vasoactive intestinal peptide, prion, neuropeptide Y and receptors, and polypeptides.

11. The compound according to any one of claims 6 to 8, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof, wherein the target protein or polypeptide to which the TBM binds is at least one selected from the group consisting of bromodomain-containing proteins, Aurora kinase A and p38 MAP kinase.

12. The compound according to any one of claims 6 to 8, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof, wherein the TBM is any one selected from the group consisting of a BRD4 inhibitor, an Aurora A inhibitor and a p38 inhibitor.

13. The TBM is a compound according to any one of claims 6 to 8, having any one chemical structure selected from the group consisting of the following Chemical Formulas 1 to 3, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate, or prodrug thereof: [Chemical formula 1] [Chemical formula 2] [Chemical formula 3]

14. The compound according to any one of claims 6 to 8, having any one chemical structure selected from the group consisting of the following chemical formulas 4 to 6, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate, or prodrug thereof. [Chemical formula 4] [Chemical formula 5] [Chemical formula 6] In the above Chemical Formulae 4 to 6, Z is each independently a direct bond, or -N(R')-, -O-, -C(R')(R'')-O-, -S-, -C(O)-, -SO-, SO ２ -, =N-O-, -O-N=, -OC(O)-, -C(O)O-, -N(R')C(O)-, -C(O)N(R')-, -OC(O)O-, -OC(O)N(R')-, -N(R')C(O)O-, -N(R')SO-, -SON(R')-, -N(R')SO ２ - and -SO ２ N(R′)—, In the above functional groups, R' and R'' are each independently hydrogen or alkyl.

15. The compound according to any one of claims 6 to 8, which is at least one selected from the group consisting of the compounds listed in Tables 5 to 15 below, or a pharma- ceutically acceptable salt, isomer, solvate, hydrate, or prodrug thereof. 【Table 5】 【Table 6】 【Table 7】 【Table 8】 【Table 9】 【Table 10】 【Table 11】 【Table 12】 【Table 13】 【Table 14】 【Table 15】

16. A pharmaceutical composition for preventing or treating a disease caused by degradation of a target protein or polypeptide, comprising as an active ingredient a compound according to any one of claims 1 to 8 or a pharma- ceutical acceptable salt, isomer, solvate, hydrate or prodrug thereof.

17. The compound according to any one of claims 6 to 8 or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof is contained as an active ingredient, A pharmaceutical composition for preventing or treating a bromodomain-containing protein-associated disease, wherein the TBM in the compound is a BRD4 inhibitor.

18. The compound according to any one of claims 6 to 8 or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof is contained as an active ingredient, A pharmaceutical composition for preventing or treating a cell proliferative disorder, wherein the TBM is an Aurora A inhibitor.

19. The compound according to any one of claims 6 to 8 or a pharma- ceutically acceptable salt, isomer, solvate, hydrate or prodrug thereof is contained as an active ingredient, A pharmaceutical composition for preventing or treating inflammation and inflammation-related diseases, wherein said TBM is a p38 inhibitor.

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