CN115745971B - PARP7 inhibitor and application thereof - Google Patents

Abstract

The present disclosure relates to a compound having PARP7 inhibitory effect as shown in formula (I), and its use and preparation method.

Description

PARP7 inhibitors and their applications

Technical Field

The present disclosure relates to the field of medicine, and in particular to a compound having PARP7 inhibitory effect, and its use and preparation method.

Background technique

Poly-ADP-ribose polymerases (PARPs) are a family of proteins that consist of 17 different members in the human body. They play important roles in different biological processes, including inflammation, viral infection and tumor cell stress. The PARP family can be divided into three categories according to catalytic activity: polyPARPs (4, including PARP1, PARP2, PARP5a, PARP5b) with multiple ADP-ribosylation target proteins, monoPARPs (12, including PARP3, PARP4, PARP6, PARP7, PARP8, PARP9, PARP10, PARP11, PARP12, PARP14, PARP15, PARP16) with single ADP-ribosylation target proteins and PARP with no catalytic activity (1, PARP13). Most of the research on PARP is focused on polyPARPs (PARP1, 2, 5a and 5b), especially PARP1/2, which has the most research on the role of DNA damage repair. DNA damage refers to the phenomenon that the DNA nucleotide sequence is permanently changed during DNA replication, resulting in changes in genetic characteristics. If DNA damage or abnormal changes in genetic information cannot be corrected, it will affect the function or survival of cells. The repair of DNA damage is a programmed, orderly, multi-stage, and precise process involving multiple factors. PARP is an important protein in the DNA repair process and is involved in a series of important cellular processes including DNA repair and genome stability maintenance. There are 4 PARP inhibitors: niraparib and talazoparib target PARP1 and PARP2; olaparib and rucaparib target PARP1, PARP2, and PARP3. In addition, olaparib (First-in-Class), represented by the PARP1/2 inhibitor developed by AstraZeneca, has become a blockbuster drug with significant anti-cancer efficacy, and its indications can be gradually expanded through combination therapy.

There are relatively few studies on monoPARPs, which are single ADP-ribosylated target proteins. As mentioned above, most members (12) of the PARP family are monoPARPs. In recent years, studies on monoPARPs have found that PARP7 plays an important role in immunity and cancer. Among them, studies suggest that PARP7 is one of the target genes of AHR. PARP7 is part of the negative feedback loop that regulates AHR activity, and AHR can regulate immune function, inflammation, etc., and play a role in cancer. It can be induced by cancer-related stress, such as harmful chemicals in tobacco. Through in-depth studies of lung cancer, esophageal cancer, and head and neck cancer, which are strongly correlated with smoking, it was found that the gene locus where PARP7 is located is amplified and highly expressed. Studies have found that PARP7 is overactive in tumor cells, and many cancer cells rely on PARP7 for cell survival. Studies have shown that PARP7 can enable cancer cells to escape immune attacks by the immune system. Inhibiting PARP7 can effectively inhibit the growth of cancer cells, restore interferon signaling, and reverse immunosuppression. In several cancer models, PARP7 inhibitors have shown persistent tumor growth inhibition and interferon signaling restoration. In summary, PARP7 plays an important role in cancer and immune response and is a potential ideal cancer target. Due to the lack of PARP7-selective chemical probes in previous studies, most of the genetic methods were used to reveal the function of PARP7, so it was impossible to distinguish between enzymatic activity and the entire protein and their relationship. In addition, the lack of mature high-throughput on-target activity screening test methods also hindered the corresponding drug development for PARP7 targets. In view of the continuous reports of PARP7-related research results in recent years, PARP7 has become a relatively ideal anti-cancer target. Therefore, it is of great significance to further study and develop new safe and effective PARP7 inhibitors. In addition, as mentioned above, in addition to playing an important role in tumors, PARP7 also plays a key role in diseases such as inflammation and viral infection. Therefore, in addition to its anti-cancer effect, PARP7 inhibitors are also expected to have potential application value in other diseases such as inflammation, immunity, and viral infection.

Summary of the invention

The object of the present disclosure is to provide a compound having PARP7 inhibitory effect or a pharmaceutically acceptable salt thereof.

The present disclosure relates to a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

in,

_n1 is an integer selected from 0 to 3; _U1 , _U2 , and _U4 are each independently selected from CH or N.

U ₅ , U ₆ , U ₇ and U ₈ are each independently selected from CH, CH ₂ , N or NH.

R ₁ is selected from hydrogen, deuterium or C _1-3 alkyl, wherein the C _1-3 alkyl is optionally substituted by one or more halogens.

Preferably, R ₁ is selected from hydrogen, deuterium or methyl, said methyl being optionally substituted by one or more halogens.

Preferably, R ₁ is selected from hydrogen, deuterium, methyl or trifluoromethyl.

_U3 is selected from -NH-, -N( _R2 )-, _-CH2- , -CH( _R2 )-, O or S.

Wherein, _R2 is selected from _C1-3 alkyl, _C3-6 cycloalkyl or _C3-6 heterocycloalkyl.

Preferably, _R2 is selected from methyl, ethyl, isopropyl, cyclopropyl, cyclohexyl.

Preferably, _R2 is selected from methyl.

Preferably, U ₃ is selected from -NH-, O, S, or -N(CH ₃ )-.

R ₃ and R ₄ are each independently selected from hydrogen, deuterium, C _1-3 alkyl, or R ₃ and R ₄ are connected to each other to form a divalent C _3-6 cycloalkyl or a 3-7 membered heterocycloalkyl containing at least one heteroatom selected from N, O or S, and the C _1-3 alkyl, C _3-6 cycloalkyl or 3-7 membered heterocycloalkyl is optionally substituted with one or more halogen, hydroxy, amino, nitro, cyano.

Preferably, R ₃ and R ₄ are each independently selected from hydrogen, deuterium, C _1-3 alkyl, or R ₃ and R ₄ are connected to each other to form a divalent C _5-6 cycloalkyl or a 5-7 membered heterocycloalkyl containing at least one heteroatom selected from N, O or S, and the C _1-3 alkyl C _3-6 cycloalkyl is optionally substituted by one to three halogens.

Preferably, R ₃ and R ₄ are each independently selected from hydrogen, deuterium, methyl, trifluoromethyl, or R ₃ and R ₄ are connected to each other to form a divalent cyclopentyl, cyclohexyl or tetrahydrofuranyl group.

_R5 is selected from hydrogen, halogen or _C1-3 alkyl.

Preferably, R ₅ is selected from hydrogen or methyl.

Preferably, R ₅ is hydrogen.

R ₆ and R _6a are each independently selected from hydrogen, halogen or C _1-3 alkyl, or R ₆ and R _6a are connected to each other to form a C _5-7 cycloalkyl, C _5-7 aryl, a 5-7 membered heterocycloalkyl or a 5-7 membered heteroaryl containing at least one heteroatom selected from N, O or S, and the C _5-8 cycloalkyl, C _5-7 aryl, 5-7 membered heterocycloalkyl or 5-7 membered heteroaryl is optionally substituted by one or more halogen, hydroxyl, cyano, carboxyl, formamide, sulfonamide, C _1-3 alkyl, C _2-6 ester, C _3-9 cycloalkyl, 3-9 membered heterocyclyl, C _5-7 aryl or 5-7 membered heteroaryl.

Preferably, R ₆ and R _6a are each independently selected from hydrogen, chlorine or methyl, or R ₆ and R _6a are connected to each other to form cyclopentyl, cyclohexyl, tetrahydropyranyl, piperidinyl, phenyl, pyridinyl, and the cyclopentyl, cyclohexyl, tetrahydropyranyl, piperidinyl, phenyl, pyridinyl are optionally substituted with one or more fluorine, chlorine, hydroxyl, cyano, carboxyl, formamide, sulfonamide, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, spirohexanyl, spiroheptanyl, spirooctanyl, octahydropentenyl, cyclopropylcyclopentanyl, cyclopropylcyclohexanyl, oxirane, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, morpholinyl, cyclopropylpiperidinyl, cyclopropylpyrrolidinyl, octahydroindolyl, octahydroindazolyl, azaspirohexanyl, oxaspirohexanyl, azaspiroheptanyl, oxaspiroheptanyl, azaspirooctanyl, oxaspirooctanyl, phenyl.

Preferably, Selected from

_R8 , _R8a , _R9 , _R9a , _R10 , _R10a , _R11 , _R11a are each independently selected from hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, _C3-9 cycloalkyl, 3-9 membered heterocyclyl or 5-6 membered heteroaryl containing at least one heteroatom selected from N, O or S, and the _C1-6 alkyl, C2-6 alkenyl, _C2-6 alkynyl, _C3-9 cycloalkyl, _3-9 membered heterocyclyl, 5-6 membered heteroaryl, _C5-7 aryl is _optionally substituted with one or more halogen, hydroxyl, cyano, carboxyl, sulfonamide, _C1-3 alkyl, _C1-3 amide, _C2-6 ester, C R 8 , R 9 , R _{10 , and R 11 are substituted by C 3-9-membered cycloalkyl, 3-9-membered heterocyclyl, C 5-7 aryl, or 5-7-membered heteroaryl, or any two of R 8 , R 9 , R 10 , and R 11 are linked to form} —(CH ₂ ) _{n 2} —, wherein n ₂ is 1, 2, or 3.

Preferably, R ₈ , R _8a , R ₉ , R _9a , R ₁₀ , R _10a , R ₁₁ , R _11a are each independently selected from hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl, vinyl, propenyl, allyl, butenyl, ethynyl, propynyl, propargyl, 2-methyl-2-propynyl, 2-butynyl, 2-pentynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, spirohexanyl, spiroheptanyl, spirooctanyl, octahydropentenyl, cyclopropylcyclopentanyl, cyclopropyl cyclohexyl, oxirane, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, morpholinyl, cyclopropylpiperidinyl, cyclopropylpyrrolidinyl, octahydroindolyl, octahydroindazolyl, azaspirohexanyl, oxaspirohexanyl, azaspiroheptanyl, oxaspiroheptanyl, azaspirooctanyl, oxaspirooctanyl, phenyl, pyrrolyl, pyrazolyl, triazolyl, imidazolyl, furanyl, isoxazolyl, pyridinyl, pyrimidinyl, triazinyl, the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl, vinyl, propenyl, allyl, butenyl, ethynyl, propynyl, propargyl, 2-methyl-2-propynyl, 2-butynyl, 2-pentynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, spirohexyl, spiroheptyl, spirooctyl, octahydropentenyl, cyclopropylcyclopentanyl, cyclopropylcyclohexyl, oxiranyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, morpholinyl, cyclopropylpiperidinyl oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, phenyl, pyrrolyl, pyrazolyl, triazolyl, imidazolyl, furanyl, isoxazolyl, pyridinyl, pyrimidinyl, triazinyl, optionally substituted by one, two or three halogens, hydroxyl, cyano, carboxyl, formamido, sulfonamido, methyl, ethyl, propyl, isopropyl, -C(O)-O-CH _{R 3} -, -C(O)-O-CH ₂ CH ₃ -, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, spirohexanyl, spiroheptanyl, spirooctanyl, octahydropentenyl, cyclopropylcyclopentanyl, cyclopropylcyclohexanyl, oxirane, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, morpholinyl, cyclopropylpiperidinyl, cyclopropylpyrrolidinyl, octahydroindolyl, octahydroindazolyl, azaspirohexanyl, oxaspirohexanyl, azaspiroheptanyl, oxaspiroheptanyl, azaspirooctanyl, oxaspirooctanyl, phenyl, pyrrolyl, pyrazolyl, triazolyl, imidazolyl, furanyl, isoxazolyl, pyridinyl, pyrimidinyl, triazinyl, or any two of R ₈ , R ₉ , R ₁₀ , and R ₁₁ are linked to each other to form -(CH ₂ ) _n2 -, wherein n ₂ is 1, 2, or 3.

Preferably, R ₈ , R _8a , R ₉ , R _9a , R ₁₀ , R _10a , R ₁₁ , R _11a are each independently selected from hydrogen, deuterium, halogen, methyl or ethyl, and the methyl or ethyl is optionally substituted by one or two cyano groups, formamide groups or -C(O)-O-CH ₂ CH ₃ -, or any two of R ₈ , R ₉ , R ₁₀ , R ₁₁ are connected to each other to form -(CH ₂ ) _n2 -, wherein n ₂ is 1, 2 or 3.

Preferably, Selected from

n ₁ R ₁₂ are the same as or different from each other, and R ₁₂ is selected from hydrogen, deuterium, halogen, cyano, C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyl, C _2-6 alkynyl. The C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyl, C _2-6 alkynyl are optionally substituted with one or more halogen, hydroxyl, C _2-6 ester, C _1-3 amide, sulfonamide.

Preferably, n ₁ R ₁₂ are the same as or different from each other, and R ₁₂ is selected from hydrogen, deuterium, fluorine, chlorine, cyano, methyl, ethyl, propyl, isopropyl, methoxy, vinyl, propenyl, allyl, butenyl, ethynyl, propynyl, propargyl, 2-methyl-2-propynyl, 2-butynyl, 2-pentynyl. The methyl, ethyl, propyl, isopropyl, methoxy, vinyl, propenyl, allyl, butenyl, ethynyl, propynyl, propargyl, 2-methyl-2-propynyl, 2-butynyl, 2-pentynyl are optionally substituted with one, two or three fluorine, hydroxyl, -C(O)-O-CH ₃ -, -C(O)-O-CH ₂ CH ₃ -, formamide, sulfonamide.

Preferably, n ₁ R ₁₂ are the same as or different from each other, and R ₁₂ is selected from hydrogen, deuterium, halogen, cyano, methyl or methoxy, and the methyl or methoxy is optionally substituted by one, two or three fluorine.

Preferably, Selected from

The present disclosure relates to a compound represented by the following formula (II) or a pharmaceutically acceptable salt thereof:

Wherein, _n1 , _U1 , _U2 , _U3 , _U4 , U5, _U6 , _U7 , _U8 , _R3 , _R4 , _R6 , _R6a , _R8 , _R8a , _R9 , _R10 and _{R12 are} as defined in the present application.

The present disclosure relates to a compound represented by the following formula (IIIa), (IIIb), (IIIc), (IIId) or (IIIe) or a pharmaceutically acceptable salt thereof:

in,

_n3 is selected from 0, 1 or 2; _n4 is selected from 0 or 1;

X ₁ , X ₂ , X ₃ , and X ₄ are each independently selected from CH or N;

_X5 is selected from _CH2 or O;

R ₁₃ is selected from hydrogen, halogen or methyl;

_n1 , _U5 , _U6 , _U8 , _R3 , _R4 , _R8 , _R8a , _R9 , _R10 and _R12 are as defined in the present application.

The present disclosure also relates to the following compounds or pharmaceutically acceptable salts thereof:

The present disclosure relates to a pharmaceutical composition comprising any one of the aforementioned compounds or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

The present disclosure also relates to the use of any one of the aforementioned compounds or pharmaceutically acceptable salts thereof, or the aforementioned pharmaceutical composition in the preparation of a drug for treating a PARP7-mediated disorder in a patient.

The present disclosure also relates to the use of any of the aforementioned compounds or pharmaceutically acceptable salts thereof, and the aforementioned pharmaceutical composition in the preparation of PARP7 inhibitors.

The present disclosure also relates to a method of inhibiting PARP7 in a patient in need thereof, comprising administering to the patient any one of the aforementioned compounds or pharmaceutically acceptable salts thereof or the aforementioned pharmaceutical composition.

The present disclosure relates to a method for inhibiting PARP7 in a biological sample, comprising contacting the biological sample with any one of the aforementioned compounds or pharmaceutically acceptable salts thereof or the aforementioned pharmaceutical composition.

The present disclosure relates to a method for treating a disorder mediated by PARP7 in a patient in need thereof, comprising administering to the patient any one of the aforementioned compounds or a pharmaceutically acceptable salt thereof or the aforementioned pharmaceutical composition.

Preferably, the PARP7-mediated disorders include, but are not limited to, cancer, immune diseases, inflammation and viral infections.

Detailed ways

According to the contents of the present disclosure, in accordance with common technical knowledge and customary means in the art, without departing from the basic technical ideas of the present disclosure, other various forms of modifications, replacements or changes may be made.

I. Definition

Unless explicitly stated otherwise, throughout the specification and claims, the term “comprise” or variations such as “include” or “comprising”, etc., will be understood to include the stated elements or components but not to exclude other elements or components.

The compounds of the present disclosure may be asymmetric, for example, having one or more stereoisomers. Unless otherwise indicated, all stereoisomers are included, such as enantiomers and diastereomers. The compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure form or racemic form. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral raw materials or chiral reagents. Racemates, diastereomers, and enantiomers are all included within the scope of the present disclosure.

The compounds of the present disclosure also include tautomeric forms. Tautomeric forms result from the exchange of a single bond with an adjacent double bond accompanied by the migration of a proton.

The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both the occurrence of said event or circumstance and the non-occurrence of said event or circumstance.

Numeric ranges herein refer to each integer in the given range. For example, "C _1-6 " means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms; "C _3-6 " means that the group can have 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms.

The term "substituted" or "substituted" means that any one or more hydrogen atoms on a particular atom or group are replaced by a substituent, as long as the valence state of the particular atom or group is normal and the substituted compound is stable. When the substituent is a keto group (i.e., =O), it means that two hydrogen atoms are replaced. Unless otherwise specified, the type and number of substituents can be any on the basis of chemical achievable.

In this disclosure, when any variable (e.g., _Rn ) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 1-5 R, the group may be optionally substituted with up to 5 R, and each occurrence of R is an independent choice. In addition, combinations of substituents and/or variants thereof are permitted only if such combinations result in stable compounds.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3 -dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 2,2-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof, etc. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate groups. Methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl and hydroxy-substituted alkyl are preferred in the present disclosure.

The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc. The alkenyl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

"Alkynyl" refers to (CH≡C-), wherein the alkynyl can be further substituted by other related groups, for example: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, cyano, nitro, phenol, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

The term "cycloalkyl" refers to a saturated monocyclic alkane substituent, the cycloalkyl ring containing at least 3 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 6 carbon atoms. Non-limiting examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

The term "heterocyclyl" or "heterocycloalkyl" refers to a saturated monocyclic cyclic hydrocarbon substituent, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (wherein m is an integer from 0 to 2), but excluding the ring portion of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Non-limiting examples of heterocyclyl include pyrrolyl, imidazolyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, etc., preferably pyrrolidinyl, morpholinyl, piperidinyl, cycloheptyl, 1,4-diazacycloheptyl and piperazinyl.

The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, cyano, nitro, chloro, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group having a conjugated π electron system, preferably 6- to 12-membered, such as phenyl and naphthyl. More preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclic or cycloalkyl ring, including benzo 5-10-membered heteroaryl, benzo 3-8-membered cycloalkyl and benzo 3-8-membered heteroalkyl, preferably benzo 5-6-membered heteroaryl, benzo 3-6-membered cycloalkyl and benzo 3-6-membered heteroalkyl, wherein the heterocyclic group is a heterocyclic group containing 1-3 nitrogen atoms, oxygen atoms, and sulfur atoms; or further comprising a three-membered nitrogen-containing fused ring containing a benzene ring.

The aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydrogen, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

The term "heteroaryl" refers to a heteroaromatic system comprising heteroatoms and carbon atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5-membered or 6-membered, such as imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidyl, thiadiazole, pyrazinyl, etc., preferably triazolyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, pyrimidyl or thiazolyl; more preferably pyrazolyl, pyrrolyl and oxazolyl.

The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydrogen, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

The term "alkoxy" refers to-O-(alkyl) and-O-(non-substituted cycloalkyl), wherein the definition of alkyl is as described above. The non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or non-substituted, and when substituted, substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfydryl, hydrogen, nitro, chloro, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

"Alkylthio-alkyl" refers to an alkylthio group attached to an alkyl group, wherein alkyl and alkylthio are as defined above; "alkylaminocarbonyl" refers to (alkyl)-N-C(O)-, wherein alkyl is as defined above; "haloalkyl" refers to an alkyl group substituted by one or more halogens, wherein alkyl is as defined above; "haloalkoxy" refers to an alkoxy group substituted by one or more halogens, wherein alkoxy is as defined above; "haloalkoxy" refers to an alkylthio group substituted by one or more halogens, wherein alkylthio is as defined above; "hydroxyalkyl" refers to an alkyl group substituted by a hydroxy group, wherein alkyl is as defined above.

In the present disclosure, "hydroxy" refers to an -OH group; "halogen" refers to fluorine, chlorine, bromine or iodine; "amino" refers to _-NH2 ; "cyano" refers to -CN; "nitro" refers to _-NO2 ; "carbonyl" refers to -C(O)-; "carboxyl" refers to -C(O)OH; "THF" refers to tetrahydrofuran; "EtOAc" refers to ethyl acetate; "MeOH" refers to methanol; "DMF" refers to N,N-dimethylformamide; "DIPEA" refers to diisopropylethylamine; "TFA" refers to trifluoroacetic acid; "MeCN" refers to acetonitrile; "DMA" refers to N,N-dimethylacetamide; "DCE" refers to 1,2-dichloroethane; "NBS" refers to N-bromosuccinimide; "NIS" refers to N-iodosuccinimide; "cbz-cr" refers to benzyl chloroformate; " _Pd2 (dba) ₃ " refers to tris(dibenzylideneacetone)dipalladium; "Dppf' refers to 1,1'-bis(diphenylphosphinoferrocene);"HATU" refers to 2-(7-oxybenzotriazolyl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; "KHMDS" refers to potassium hexamethyldisilazide; "LiHMDS" refers to lithium bis(trimethylsilylamide); "MeLi" refers to methyllithium; "n-BμLi" refers to n-butyllithium; and "NaBH(OAc) ₃ " refers to sodium triacetoxyborohydride.

In the present disclosure, different terms such as “X is selected from A, B, or C”, “X is selected from A, B and C”, “X is A, B or C”, “X is A, B and C” all express the same meaning, that is, X can be any one or more of A, B, C.

The hydrogen atoms described in the present disclosure may all be replaced by their isotope deuterium.

"Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1-3 hydrogen atoms in the group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the skilled person can determine (by experiment or theory) possible or impossible substitutions without undue effort.

In this disclosure, Refers to the chemical bond connection.

Drug or drug composition

The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness of the free acids and bases of a particular compound without adverse biological effects, such as acid (including organic acids and inorganic acids) addition salts or base addition salts (including organic bases and inorganic bases).

The pharmaceutically acceptable salts of the present disclosure can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. In general, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, via the free acid or base form of these compounds with a stoichiometric amount of an appropriate base or acid to prepare.

The medicaments or pharmaceutical compositions of the present disclosure can be administered orally, topically, parenterally or mucosally (e.g., buccally, by inhalation or rectally) in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers. It is generally desirable to use the oral route. The active agent can be administered orally in the form of capsules, tablets, etc. (see Remington: The Science and Practice of Pharmacy, 20th Edition).

For oral administration in the form of tablets or capsules, the active drug component can be mixed with non-toxic, pharmaceutically acceptable excipients such as binders (e.g., pregelatinized corn starch, polyvinyl pyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose, sucrose, glucose, mannitol, sorbitol and other reducing and non-reducing sugars, microcrystalline cellulose, calcium sulfate or dibasic calcium phosphate); lubricants (e.g., magnesium stearate, talc or silica, stearic acid, sodium stearyl fumarate, glyceryl behenate, calcium stearate, etc.); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulfate), coloring and flavoring agents, gelatin, sweeteners, natural and synthetic gums (such as acacia, tragacanth or alginates), buffer salts, carboxymethylcellulose, polyethylene glycol, waxes, and the like. For oral administration in liquid form, the drug component can be combined with a pharmaceutically acceptable inert carrier (e.g., ethanol, glycerol, water), an anti-settling agent (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats), an emulsifier (e.g., lecithin or gum arabic), a non-aqueous carrier (e.g., almond oil, oily esters, ethanol or fractionated vegetable oils), a preservative (e.g., methyl p-hydroxybenzoate or propyl p-hydroxybenzoate or sorbic acid), etc. Stabilizers such as antioxidants (BHA, BHT, propyl citric acid, sodium ascorbate, citric acid) can also be added to stabilize the dosage form.

Tablets containing the active compound can be coated by methods well known in the art. The compositions of the present disclosure containing the compound of formula I as the active compound can also introduce beads, microspheres or microcapsules, such as those constructed from polyglycolic acid/lactic acid (PGLA). Liquid preparations for oral administration can take the form of, for example, solutions, syrups, emulsions or suspensions or they can be presented as dry products reconstituted with water or other suitable excipients before use. Preparations for oral administration can be suitably formulated to release the active compound in a controlled or delayed manner.

The medicine or pharmaceutical composition of the present disclosure can be delivered parenterally, i.e., administered intravenously (i.v.), intracerebroventricularly (i.c.v.), subcutaneously (s.c.), intraperitoneally (i.p.), intramuscularly (i.m.), subcutaneously (s.d.) or intradermally (i.d.), by direct injection, for example, by bolus injection or continuous infusion. The formulation for injection can be presented in unit dosage form, for example, in an ampoule or multi-dose container with an added preservative. The composition can be in the form of an excipient, in the form of a suspension, solution or emulsion in an oil or aqueous vehicle, and can include formulation agents such as anti-settling agents, stabilizers and/or dispersants. Alternatively, the active ingredient can be reconstituted with a suitable carrier (e.g., sterile pyrogen-free water) before use in powder form.

The medicaments or pharmaceutical compositions of the present disclosure may also be formulated for rectal administration, for example, as a suppository or retention enema (eg, containing a conventional suppository base such as cocoa butter or other glycerides).

The term "treating" includes inhibiting, alleviating, preventing or eliminating one or more symptoms or side effects associated with the disease, condition or disorder being treated.

The terms "reduce," "inhibit," "mitigate," or "reduce" are used relative to a control. One skilled in the art will readily determine the appropriate control for each experiment. For example, a reduced response in a subject or cell treated with a compound is compared to a response in a subject or cell not treated with the compound.

As used herein, the term "effective amount" or "therapeutically effective amount" refers to a dosage sufficient to treat, suppress or alleviate one or more symptoms of the disease state being treated or otherwise provide a desired pharmacological and/or physiological effect. The exact dosage will vary according to a variety of factors, such as variables that the subject depends on (e.g., age, immune system health, etc.), the disease or illness, and the treatment administered. The effect of an effective amount can be relative to a control. These controls are known in the art and discussed herein, and can be, for example, the condition of a subject before or without the administration of a drug or drug combination, or in the case of a drug combination, the combined effect can be compared to the effect of administering only one drug.

The term "excipient" is used herein to include any other compound that may be contained in or on the microparticles that is not a therapeutic or biologically active compound. Thus, an excipient should be pharmaceutically or biologically acceptable or relevant, e.g., an excipient is generally non-toxic to a subject. An "excipient" includes a single such compound, and is also intended to include a plurality of compounds.

The term "pharmaceutical composition" means a composition comprising the compound described in the present disclosure or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable ingredient selected from the following depending on the mode of administration and the nature of the dosage form, including but not limited to: carriers, diluents, adjuvants, excipients, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, dispersants, temperature-sensitive materials, temperature regulators, adhesives, stabilizers, suspending agents, etc.

Uses and treatments

The terms "patient," "subject," "individual," and the like are used interchangeably herein and refer to any animal or cell thereof amenable to the methods described herein, whether in vitro or in situ. In some non-limiting embodiments, the patient, subject, or individual is a human.

[00136] According to the methods of the present disclosure, the compounds or compositions may be administered using any amount and any route of administration effective for treating or lessening the severity of a disease associated with PARP.

The present disclosure relates to a method of inhibiting PARP in a biological sample, comprising the step of contacting the biological sample with a compound of the present disclosure or a composition comprising the compound.

The term "biological sample" includes, but is not limited to, cell cultures or extracts thereof; biopsy material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears or other body fluids or extracts thereof. Inhibition of enzymes in biological samples can be used to achieve a variety of purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, bioanalysis, gene expression studies, and identification of biological targets.

The method of inhibiting PARP in a patient disclosed herein comprises the step of administering to the patient a compound disclosed herein or a composition comprising the compound.

The provided compounds are PARP inhibitors and are therefore useful for treating one or more conditions associated with PARP activity. Thus, in certain embodiments, the present disclosure provides a method for treating a PARP-mediated condition comprising the step of administering a compound of the present disclosure or a pharmaceutically acceptable composition thereof to a patient in need thereof.

As used herein, the term "PARP-mediated" disorder, disease and/or condition as used herein means any disease or other deleterious condition in which PARP or a mutant thereof is known to play a role. Thus, another embodiment of the present disclosure is directed to treating or lessening the severity of one or more diseases in which PARP or a mutant thereof is known to play a role.

The present disclosure provides a method for treating one or more disorders, diseases, and/or conditions, wherein the disorder, disease, or condition is a proliferative disorder, such as cancer, an inflammatory disorder, or a viral infection.

In certain embodiments, the present disclosure provides a method of treating cancer or another proliferative disorder, comprising administering a compound or composition of the present disclosure to a patient suffering from cancer or another proliferative disorder. In certain embodiments, the method of treating cancer or another proliferative disorder comprises administering a compound and composition of the present disclosure to a mammal. In certain embodiments, the mammal is a human.

As used herein, the terms "inhibit cancer" and "inhibit cancer cell proliferation" refer to inhibiting the growth, division, maturation or survival of cancer cells, and/or causing cancer cell death by cytotoxicity, nutrient depletion or induction of apoptosis, either individually or collectively with other cancer cells.

Examples of tissues containing cancer cells whose proliferation is inhibited by the compounds and compositions described herein and for which the methods described herein are applicable include, but are not limited to, breast, prostate, brain, blood, bone marrow, liver, pancreas, epidermis, kidney, colon, ovary, lung, testis, penis, thyroid, parathyroid, pituitary, thymus, retina, uvea, conjunctiva, spleen, head, neck, trachea, gall bladder, rectum, salivary glands, adrenal glands, throat, esophagus, lymph nodes, sweat glands, sebaceous glands, muscle, heart, and stomach.

The cancer treated by the compounds or compositions of the present disclosure is melanoma, liposarcoma, lung cancer, breast cancer, prostate cancer, leukemia, kidney cancer, esophageal cancer, brain cancer, lymphoma or colon cancer. In certain embodiments, the cancer is primary effusion lymphoma (PEL).

The compounds of the present disclosure can be used to treat a proliferative disease selected from the group consisting of a benign or malignant tumor, carcinoma of the brain, kidney, liver, adrenal gland, bladder, breast, stomach, gastric tumor, ovary, colon, rectum, prostate, pancreas, lung, vagina, cervix, testis, genitourinary tract, esophagus, larynx, skin, bone or thyroid; a sarcoma, glioblastoma, neuroblastoma, multiple myeloma or gastrointestinal cancer (particularly colon cancer or colorectal adenoma) or a tumor of the neck and head, an epidermal hyperplasia Hyperplasia, psoriasis, prostatic hyperplasia, neoplasia, neoplasia of epithelial character, adenoma, adenocarcinoma, keratoacanthoma, epidermoid carcinoma, large cell carcinoma, non-small cell lung cancer, Hodgkins and non-Hodgkin's lymphoma, breast cancer, follicular carcinoma, undifferentiated tumor, papillary carcinoma, seminoma, melanoma, MYD88 driven disorder, DLBCL, ABCDLBCL, IL-1 driven disorder and remitting or indolent multiple myeloma or leukemia.

Cancers described in the present disclosure include, but are not limited to, leukemias (e.g., acute leukemias, acute lymphocytic leukemias, acute myeloid leukemias, acute myeloblastic leukemias, acute promyelocytic leukemias, acute myelomonocytic leukemias, acute monocytic leukemias, acute erythroleukemias, chronic leukemias, chronic myelocytic leukemias, chronic lymphocytic leukemias), polycythemia vera, lymphomas (e.g., Hodgkin's disease or non-Hodgkin's disease), Waldenstrom's macroglobulinemia, multiple myeloma, heavy chain disease, and solid tumors, such as sarcomas and carcinomas (e.g., fibrosarcomas, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphoendotheliosarcoma, synovioma, mesothelioma, Ewing's tumors, g′stumor), leiomyosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct cancer, choriocarcinoma, seminoma, embryonal tumor, Wilms tumor, cervical cancer, uterine cancer, testicular cancer, lung cancer, small cell lung cancer, bladder cancer, epithelial cancer, glioma, astrocytoma, glioblastoma multiforme (GBM, also known as glioblastoma), medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma, melanoma, neuroblastoma and retinoblastoma.

In some embodiments, the cancer is a glioma, an astrocytoma, a glioblastoma multiforme (GBM, also known as a glioblastoma), a medulloblastoma, a craniopharyngioma, an ependymoma, a pinealoma, an hemangioblastoma, an acoustic neuroma, an oligodendroglioma, a schwannoma, a neurofibrosarcoma, a meningioma, a melanoma, a neuroblastoma, or a retinoblastoma.

In some specific embodiments, the cancer is an acoustic neuroma, an astrocytoma (e.g., grade I-pilocytic astrocytoma, grade II-low-grade astrocytoma, grade III-polymorphic astrocytoma, or grade IV-glioblastoma (GBM)), a chordoma, a CNS lymphoma, a craniopharyngioma, a brainstem glioma, an ependymoma, a mixed glioma, an optic nerve glioma, a subependymoma, a medulloblastoma, a meningioma, a metastatic brain tumor, an oligodendroglioma, a pituitary tumor, a primary neuroectodermal (PNET) tumor, or a schwannoma. In some embodiments, the cancer is a type that is more common in children than in adults, such as a brainstem glioma, a craniopharyngioma, an ependymoma, a juvenile pilocytic astrocytoma (JPA), a medulloblastoma, an optic nerve glioma, a pineal tumor, a primary neuroectodermal tumor (PNET), or a rhabdoid tumor. In some embodiments, the patient is an adult patient. In some embodiments, the patient is a child or a pediatric patient.

In another specific embodiment, cancers include, but are not limited to, mesothelioma, hepatobiliary (liver and bile duct), bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cutaneous or intraocular melanoma, ovarian cancer, colon cancer, rectal cancer, cancer of the anal region, gastric cancer, gastrointestinal tract (stomach, colorectal and duodenum), uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, Adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, testicular cancer, chronic or acute leukemia, chronic myeloid leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvis cancer, non-Hodgkin's lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, adrenocortical carcinoma, gallbladder cancer, multiple myeloma, bile duct cancer, fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one or more of the foregoing cancers.

In some embodiments, the cancer is selected from hepatocellular carcinoma, ovarian cancer, ovarian epithelial cancer, or fallopian tube cancer; papillary serous cystadenocarcinoma or uterine serous papillary carcinoma (UPSC); prostate cancer; testicular cancer; gallbladder cancer; cholangiohepatoma; synovial sarcoma of soft tissue and bone; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing's sarcoma; pleomorphic thyroid cancer; adrenocortical adenoma; pancreatic cancer; pancreatic duct cancer or pancreatic cancer; gastrointestinal tract/gastric (GIST) cancer; lymphoma; squamous cell carcinoma of the head and neck (SCCHN); salivary gland cancer; glioma or brain cancer; neurofibroma-1-associated malignant peripheral nerve sheath tumor (MPNST); Waldenstrom's macroglobulinemia; or medulloblastoma.

The term "primary tumor" is relative to secondary tumor. Primary tumor refers to a tumor that first appears in a certain part of the body such as the lungs, liver, intestines, head, or skin. It can be called primary lung cancer, primary liver cancer, primary intestinal cancer, etc.

The term "inflammatory disease" includes such autoimmune, allergic and inflammatory disorders, for example selected from arthritis, ankylosing spondylitis, inflammatory bowel disease, ulcerative colitis, gastritis, pancreatitis, Crohn's disease, celiac disease, multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis, rheumatic fever, gout, organ or transplant rejection, acute or chronic graft-versus-host disease, chronic allograft rejection, Behcet's disease, uveitis, psoriasis, dermatitis, atopic dermatitis, dermatomyositis, myasthenia gravis, Grave's disease, Hashimoto's thyroiditis, Sjogren's syndrome, and blistering disorders (e.g. pemphigus vulgaris), antibody-mediated vasculitis syndromes, including ANCA-associated vasculitis, purpura, and immune complex vasculitis (cancer or infection primary or secondary). The allergic disorder may be selected in particular from contact dermatitis, celiac disease, asthma, hypersensitivity to house dust mites, pollens and related allergens, berylliosis. The respiratory disorder may be selected from, inter alia, asthma, bronchitis, chronic obstructive pulmonary disease (COPD), cystic fibrosis, pulmonary edema, pulmonary embolism, pneumonia, pulmonary sarcoidosis, silicosis, pulmonary fibrosis, respiratory failure, acute respiratory distress syndrome, primary pulmonary hypertension and emphysema, among others.

The term "viral infection" includes but is not limited to retroviral infection, hepatitis virus infection, COVID-19 novel coronavirus infection, Zika virus infection, dengue virus infection, etc.

Combination therapy approach

The present disclosure provides a combination therapy using compounds as described in the present disclosure and other therapeutic drugs. The term "combination therapy" used in the present disclosure includes the administration of these agents in a sequential manner, i.e., each therapeutic agent is administered at different times, and the administration of these therapeutic agents, or at least two agents, is substantially performed simultaneously. The order of each agent, or substantially simultaneous administration, can be affected by any appropriate route, including, but not limited to, oral routes, intravenous routes, intramuscular routes, subcutaneous routes, and direct absorption through mucosal tissue. The agents can be administered by the same route or different routes. For example, the first agent can be administered orally, and the second agent can be administered intravenously. In addition, the selected combination can be administered by intravenous injection, and the other agents of the combination can be administered orally. Alternatively, for example, two or more agents can be administered by intravenous or subcutaneous injection.

II. Embodiment

The present disclosure is further explained below with reference to examples. The description of specific exemplary embodiments of the present disclosure is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present disclosure to the precise form disclosed, and it is clear that many changes and variations can be made according to the teachings of the present disclosure specification. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present disclosure and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present disclosure and various different selections and changes.

Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

Experimental instruments and methods:

Instruments and reagents:

NMR was measured using an Agilent 400MR DD2 nuclear magnetic spectrometer, with deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated methanol (CD ₃ OD) and deuterated chloroform (CDCl ₃ ), and tetramethylsilane (TMS) as the internal standard. LC-MS: Agilent 1260 Infinity II-InfinityLab LC/MSD mass spectrometer. HPLC: Agilent 1260 InfinityII high pressure liquid chromatograph (Sunfire C18 5um 150x4.6mm column). Thin layer chromatography silica gel plate used HSGF254 silica gel plate (Yantai Jiangyou Silica Gel Development Co., Ltd.), with a specification of 0.9mm-1mm. TLC silica gel plate used GF254 silica gel plate (Yucheng Chemical (Shanghai) Co., Ltd.), with a specification of 0.2mm-0.25mm. Column chromatography: carrier 300-400 mesh silica gel (Qingdao Hailang Silica Gel Desiccant Co., Ltd.), Flash column (Aiger Finomei Claricep Flash amorphous silica gel purification column). Reagents used were 1,1-cyclopropyldicarboxylic acid monomethyl ester, 1,1-cyclopropane dimethanol and Pd/C (Shanghai Haohong Biopharmaceutical Technology Co., Ltd.), 4N hydrochloric acid dioxane solution (Shanghai Kaiman Chemical Technology Co., Ltd.), and other reagents and starting materials were purchased from Shanghai Bid Reagent Co., Ltd. or synthesized using methods known in the art.

Unless otherwise specified, all reactions disclosed herein are carried out under continuous magnetic stirring, in dry nitrogen or argon, with dry solvents and reaction temperatures in degrees Celsius.

Table 1 Intermediate List

Preparation method of intermediate

(1) Preparation of Intermediate I-1: 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-1)

Step 1: Preparation of tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-1a)

2-Chloro-5-(trifluoromethyl)pyrimidine (1825 mg, 10 mmol) was dissolved in N-methylpyrrolidone (30 ml), and tert-butyl piperazine-1-carboxylate (1862 mg, 10 mmol) and potassium carbonate (2764 mg, 20 mmol) were added, and the mixture was reacted at 80°C for 1 hour. Water (100 ml) was added, and the solid was collected by filtration and dried in vacuo to obtain the target product tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-1a, 2.9 g, yield 87%).

Step 2: Preparation of 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-1)

Dissolve 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-1a, 332 mg, 1 mmol) in 4M hydrochloric acid ethyl acetate (5 ml) and react at room temperature for 2 hours. Filter the reaction solution and wash the solid with petroleum ether to obtain the target product 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-1, 289 mg, yield 94%). ESI [M+H] ⁺ = 233.1. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.48 (s, 2H), 8.78 (s, 2H), 4.06 (s, 4H), 3.18 (s, 4H).

(2) Intermediate I-2: Preparation of 5-methyl-2-(piperazin-1-yl)pyrimidine hydrochloride (I-2)

Step 1: Preparation of tert-butyl 4-(5-methylpyrimidin-2-yl)piperazine-1-carboxylate (I-2a)

2-Chloro-5-methylpyrimidine (500 mg, 3.9 mmol) was dissolved in N-methylpyrrolidone (10 ml), and piperazine-1-carboxylic acid tert-butyl ester (727 mg, 3.9 mmol) and potassium carbonate (1.08 g, 7.8 mmol) were added, and the mixture was reacted at 80°C for 8 hours. Saturated brine (20 ml) was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the target product, 4-(5-methylpyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-2a, 1.08 g, yield 100.0%). ESI[M+H] ⁺ =279.2, ESI[M-56+H] ⁺ =233.1, ESI[M-100+H] ⁺ =179.2

Step 2: Preparation of 5-methyl-2-(piperazin-1-yl)pyrimidine hydrochloride (I-2)

Dissolve tert-butyl 4-(5-methylpyrimidin-2-yl)piperazine-1-carboxylate (I-2a, 1.08 g, 3.88 mmol) in ethyl acetate (10 ml), add 4M hydrochloric acid dioxane solution (30 ml), and react at room temperature for 2 hours. Collect the precipitated solid by filtration, wash with ethyl acetate to obtain the target product 5-methyl-2-(piperazin-1-yl)pyrimidine hydrochloride (I-2, 645 mg, yield 77.6%).

(3) Preparation of Intermediate I-3: 3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane hydrochloride (I-3)

Step 1: Preparation of tert-butyl 3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane-8-carboxylate (I-3a)

Dissolve tert-butyl 3,8-diazacyclo[3.2.1]octane-8-carboxylate (200 mg, 0.942 mmol) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyrimidine (172 mg, 0.942 mmol) and potassium carbonate (260 mg, 1.88 mmol), and react at 80°C for 5 hours. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product tert-butyl 3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane-8-carboxylate (I-3a, 312 mg, yield 92.6%). ESI[M-56+H] ⁺ =303.1.

Step 2: Preparation of 3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane hydrochloride (I-3)

3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (I-3a, 312 mg, 0.871 mmol) was dissolved in ethyl acetate (5 ml), and a 4M hydrochloric acid dioxane solution (10 ml) was added, and the mixture was reacted at room temperature for 3 hours. The precipitated solid was collected by filtration and washed with ethyl acetate to obtain the target product 3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane hydrochloride (I-3, 232 mg, yield 90.6%).

(4) Intermediate I-4: Preparation of 5-fluoro-2-(piperazin-1-yl)pyrimidine hydrochloride (I-4)

Step 1: Preparation of tert-butyl 4-(5-fluoropyrimidin-2-yl)piperazine-1-carboxylate (I-4a)

Dissolve tert-butyl piperazine-1-carboxylate (200 mg, 1.07 mmol) in N-methylpyrrolidone (5 ml), add 2-chloro-5-fluoropyrimidine (143 mg, 1.07 mmol) and potassium carbonate (296 mg, 2.15 mmol), and react at 100°C for 5 hours. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product tert-butyl 4-(5-fluoropyrimidin-2-yl)piperazine-1-carboxylate (I-4a, 211 mg, yield 69.6%). ESI[M-56+H] ⁺ =227.1, ESI[M-100+H] ⁺ =183.2.

Step 2: Preparation of 5-fluoro-2-(piperazin-1-yl)pyrimidine hydrochloride (I-4)

Dissolve tert-butyl 4-(5-fluoropyrimidin-2-yl)piperazine-1-carboxylate (I-4a, 211 mg, 0.748 mmol) in ethyl acetate (4 ml), add 4M hydrochloric acid dioxane solution (8 ml), and react at room temperature for 3 hours. Collect the precipitated solid by filtration, wash with ethyl acetate to obtain the target product 5-fluoro-2-(piperazin-1-yl)pyrimidine hydrochloride (I-4, 184 mg, yield 112.9%).

(5) Intermediate I-5: Preparation of 5-methoxy-2-(piperazin-1-yl)pyrimidine hydrochloride (I-5)

Step 1: Preparation of tert-butyl 4-(5-methoxypyrimidin-2-yl)piperazine-1-carboxylate (I-5a)

Dissolve tert-butyl piperazine-1-carboxylate (200 mg, 1.07 mmol) in N-methylpyrrolidone (5 ml), add 2-chloro-5-methoxypyrimidine (155 mg, 1.07 mmol) and potassium carbonate (296 mg, 2.15 mmol), and react at 100°C for 5 hours. Add ethyl acetate (20 ml) to dilute, wash with saturated brine (20 ml x 4), dry with anhydrous magnesium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain the target product tert-butyl 4-(5-methoxypyrimidin-2-yl)piperazine-1-carboxylate (I-5a, 265 mg, yield 83.9%). ESI[M+H] ⁺ =295.1, ESI[M-56+H] ⁺ =239.0, ESI[M-100+H] ⁺ =195.2

Step 2: Preparation of 5-methoxy-2-(piperazin-1-yl)pyrimidine hydrochloride (I-5)

Dissolve tert-butyl 4-(5-methoxypyrimidin-2-yl)piperazine-1-carboxylate (I-5a, 265 mg, 0.9 mmol) in ethyl acetate (5 ml), add 4M hydrochloric acid dioxane solution (5 ml), and react at room temperature for 3 hours. Collect the precipitated solid by filtration, wash with ethyl acetate to obtain the target product 5-methoxy-2-(piperazin-1-yl)pyrimidine hydrochloride (I-5, 205 mg, yield 99.0%).

(6) Intermediate I-6: Preparation of (S)-2-(2-methylpiperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-6)

Step 1: Preparation of (S)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-6a)

Dissolve (S)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (200 mg, 1 mmol) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyrimidine (183 mg, 1 mmol) and potassium carbonate (276 mg, 2 mmol), and react at 100°C for 2 hours. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product (S)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-6a, 276 mg, yield 79.8%). ESI[M-56+H] ⁺ =291.0, ESI[M-100+H] ⁺ =247.1

Step 2: Preparation of (S)-2-(2-methylpiperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-6)

Dissolve (S)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-6a, 276 mg, 0.797 mmol) in ethyl acetate (5 ml), add 4M hydrochloric acid dioxane solution (5 ml), and react at room temperature for 3 hours. Collect the precipitated solid by filtration, wash with ethyl acetate to obtain the target product (S)-2-(2-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-6, 220 mg, yield 97.8%).

(7) Intermediate I-7: Preparation of 1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-7)

Step 1: Preparation of tert-butyl 4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (I-7a)

Dissolve tert-butyl piperazine-1-carboxylate (200 mg, 1.07 mmol) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyridine (195 mg, 1.07 mmol) and potassium carbonate (296 mg, 2.15 mmol), and react at 100°C for 3 hours. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product tert-butyl 4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (I-7a, 155 mg, yield 43.7%).

Step 2: Preparation of 1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-7)

Dissolve tert-butyl 4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (I-7a, 155 mg, 0.468 mmol) in ethyl acetate (4 ml), add 4M hydrochloric acid dioxane solution (4 ml), and react at room temperature for 2 hours. Collect the precipitated solid by filtration, wash with ethyl acetate to obtain the target product 1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-7, 131 mg, yield 104%). ESI[M+H] ⁺ =232.1

(8) Intermediate I-8: Preparation of 2-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazacyclo[2.2.1]heptane hydrochloride (I-8)

Step 1: Preparation of tert-butyl 5-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazacyclo[2.2.1]heptane-2-carboxylate (I-8a)

Dissolve tert-butyl 2,5-diazacyclo[2.2.1]heptane-2-carboxylate (125 mg, 0.63 mmol) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyrimidine (115 mg, 0.63 mmol) and potassium carbonate (174 mg, 1.26 mmol), and react at 100°C for 3 hours. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product tert-butyl 5-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazacyclo[2.2.1]heptane-2-carboxylate (I-8a, 189 mg, yield 87.1%).

Step 2: Preparation of 2-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazacyclo[2.2.1]heptane hydrochloride (I-8)

Dissolve 5-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazacyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester (I-8a, 189 mg, 0.549 mmol) in ethyl acetate (5 ml), add 4M hydrochloric acid dioxane solution (5 ml), and react at room temperature for 2 hours. Collect the precipitated solid by filtration, wash with ethyl acetate to obtain the target product 2-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazacyclo[2.2.1]heptane hydrochloride (I-8, 158 mg, yield 102.6%).

(9) Intermediate I-9: Preparation of 2-(3,3-dimethylpiperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-9)

Step 1: Preparation of tert-butyl 2,2-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-9a)

Dissolve tert-butyl 2,2-dimethylpiperazine-1-carboxylate (200 mg, 0.933 mmol) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyrimidine (171 mg, 0.933 mmol) and potassium carbonate (258 mg, 1.87 mmol), and react at 100°C for 2 hours. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product tert-butyl 2,2-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-9a, 261 mg, yield 77.7%).

Step 2: Preparation of 2-(3,3-dimethylpiperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-9)

Dissolve tert-butyl 2,2-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-9a, 261 mg, 0.725 mmol) in ethyl acetate (5 ml), add 4M hydrochloric acid dioxane solution (5 ml), and react at room temperature for 2 hours. Collect the precipitated solid by filtration, wash with ethyl acetate to obtain the target product 2-(3,3-dimethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-9, 225 mg, yield 104.7%).

(10) Preparation of Intermediate I-10: 5-(Piperazin-1-yl)pyrazine-2-carbonitrile Hydrochloride (I-10)

Step 1: Preparation of tert-butyl 4-(5-cyanopyrazine-2-yl)piperazine-1-carboxylate (I-10a)

Dissolve tert-butyl piperazine-1-carboxylate (134 mg, 0.719 mmol) in N-methylpyrrolidone (5 ml), add 5-chloropyrazine-2-carbonitrile (100 mg, 0.719 mmol) and potassium carbonate (198 mg, 1.44 mmol), and react at 100°C for 3 hours. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product tert-butyl 4-(5-cyanopyrazin-2-yl)piperazine-1-carboxylate (I-10a, 177 mg, yield 85.5%). ESI[M+H] ⁺ =290.1, ESI[2M+H] ⁺ =579.5, ESI[M-56+H] ⁺ =234.1, ESI[M-100+H] ⁺ =190.1

Step 2: Preparation of 5-(piperazine-1-yl)pyrazine-2-carbonitrile hydrochloride (I-10)

Dissolve tert-butyl 4-(5-cyanopyrazin-2-yl)piperazine-1-carboxylate (I-10a, 177 mg, 0.612 mmol) in ethyl acetate (4 ml), add 4M hydrochloric acid dioxane solution (4 ml), and react at room temperature for 3 hours. Collect the precipitated solid by filtration, wash with ethyl acetate to obtain the target product 5-(piperazin-1-yl)pyrazine-2-carbonitrile hydrochloride (I-10, 122 mg, yield 88.4%).

(11) Preparation of Intermediate I-11: 2-(Piperazine-1-yl)pyrimidine-5-carbonitrile hydrochloride (I-11)

Step 1: Preparation of tert-butyl 4-(5-cyanopyrimidin-2-yl)piperazine-1-carboxylate (I-11a)

Dissolve tert-butyl piperazine-1-carboxylate (200 mg, 1.07 mmol) in N-methylpyrrolidone (5 ml), add 2-chloropyrimidine-5-carbonitrile (150 mg, 1.07 mmol) and potassium carbonate (296 mg, 2.15 mmol), and react at 100°C for 3 hours. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product tert-butyl 4-(5-cyanopyrimidin-2-yl)piperazine-1-carboxylate (I-11a, 248 mg, yield 79.9%).

Step 2: Preparation of 2-(piperazine-1-yl)pyrimidine-5-carbonitrile hydrochloride (I-11)

Dissolve tert-butyl 4-(5-cyanopyrimidin-2-yl)piperazine-1-carboxylate (I-11a, 248 mg, 0.858 mmol) in ethyl acetate (5 ml), add 4M hydrochloric acid dioxane solution (5 ml), and react at room temperature for 3 hours. Collect the precipitated solid by filtration, wash with ethyl acetate to obtain the target product 2-(piperazin-1-yl)pyrimidine-5-carbonitrile hydrochloride (I-11, 230 mg, yield 119.2%).

(12) Intermediate I-12: Preparation of (S)-2-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-12)

Step 1: Preparation of (S)-3-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-12a)

Dissolve (S)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (200 mg, 1 mmol) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyridine (181 mg, 1 mmol) and cesium carbonate (652 mg, 2 mmol), and react overnight for 120 min. Add water (5 ml), extract with ethyl acetate, wash the organic phase with saturated brine, dry over anhydrous magnesium sulfate, filter, concentrate the filtrate under reduced pressure, and purify with a Flash column (ethyl acetate: petroleum ether = 5% to 20%) to obtain the target product (S)-3-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-12a, 57 mg, yield 16.5%). ESI[M+H] ⁺ = 346.1

Step 2: Preparation of (S)-2-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-12)

Dissolve (S)-3-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-12a, 72 mg, 0.208 mmol) in 4M hydrochloric acid ethyl acetate solution (2 ml) and react at room temperature for 1.5 hours. After concentration under reduced pressure, the target product (S)-2-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-12, 66 mg, yield 112.6%) was obtained.

(13) Intermediate I-13: Preparation of (S)-2-(2-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrazine hydrochloride (I-13)

Step 1: Preparation of (S)-tert-butyl 3-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carboxylate (I-13a)

Dissolve (S)-3-methylpiperazine-1-carboxylic acid tert-butyl ester (110 mg, 0.55 mmol) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyrazine (100 mg, 0.55 mmol) and potassium carbonate (151 mg, 1.1 mmol), and react at 100°C for 3 hours. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product (S)-3-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-13a, 80 mg, yield 42.1%).

Step 2: Preparation of (S)-2-(2-methylpiperazine-1-yl)-5-(trifluoromethyl)pyrazine hydrochloride (I-13)

Dissolve (S)-3-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-13a, 80 mg, 0.231 mmol) in 4M hydrochloric acid ethyl acetate solution (4 ml) and react at room temperature for 1.5 hours. After concentration under reduced pressure, the target product (S)-2-(2-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrazine hydrochloride (I-13, 63 mg, yield 96.9%) was obtained.

(14) Intermediate I-14: Preparation of 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-14)

Step 1: Preparation of 4,5-dibromo-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-14a)

Under ice bath, 4,5-dibromopyridin-3(2H)-one (20g, 78.78mmol) was dissolved in NN dimethylformamide (200mL), 60% sodium hydride (4.0g, 94.54mmol) was slowly added, and stirred at room temperature for 1 hour, 2-(trimethylsilyl)ethoxymethyl chloride (14.4g, 86.66mmol) was added dropwise under ice bath, and after the addition was completed, the reaction solution was stirred at room temperature for 2 hours. Under ice bath, the reaction solution was quenched with water (200mL), extracted with ethyl acetate (100mL*3), and the organic phase was washed with brine (100mLx3), dried over anhydrous magnesium sulfate, filtered and concentrated. The crude product was purified by silica gel chromatography (petroleum ether: ethyl acetate = 5: 1) to give 4,5-dibromo-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-14a, 18.6 g, yield 61.46%). ESI [M+H] ⁺ = 355.1, ESI [M+3H] ⁺ = 357.1

Step 2: Preparation of 4-bromo-5-chloro-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-14b)

Lithium chloride (1.93 g, 45.30 mmol) was added to a solution of compound 4,5-dibromo-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-14a, 17.4 g, 45.30 mmol) in N-methylpyrrolidone (100 mL), and the resulting solution was stirred at 95°C for 4 h. Then 250 mL of water was added to the reaction, and the mixture was extracted with 3,50 mL of ethyl acetate, and the organic layers were combined. The organic layer was washed with 3×250 mL of brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product of 4-bromo-5-chloro-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-14b, yield 84.49%) was directly used in the next step. ESI[M+H] ⁺ =311.2, ESI[M+3H] ⁺ =313.2

Step 3: Preparation of 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-14)

To a solution of compound 4-bromo-5-chloro-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-14b, 13 g, 38.27 mmol) in N-methylpyrrolidone (80 mL) was added iodide (1.47 g, 7.66 mmol) at room temperature, followed by dropwise addition of compound 2,2-difluoro-2-(fluorosulfonyl)acetic acid methyl ester (22.17 g, 114.8 mmol). The resulting solution was stirred at 80° C. for 2 hours. The reaction was quenched by adding 250 mL of water, quenched with 3, and extracted with ethyl acetate. The organic layers were combined and washed with 3×250 mL of brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (ethyl acetate: petroleum ether = 1: 100) to give compound 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-14, 9.4 g, yield 53.95%). ESI[2M+23] ⁺ = 679.1

(15) Intermediate I-15: Preparation of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (I-15)

Step 1: Preparation of (S)-2-((tert-butyloxycarbonyl)amino)propyl 4-toluenesulfonate (I-15a)

Under ice bath, triethylamine (7.94 ml, 57.06 mmol), 4-dimethylaminopyridine (6.98 g, 57.06 mmol) and p-toluenesulfonyl chloride (8.16 g, 42.80 mmol) were added to a solution of tert-butyl (S)-(1-hydroxypropane-2-yl)carbamate (5.0 g, 28.53 mmol) in dichloromethane (100 ml), and the resulting solution was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (petroleum ether: ethyl acetate = 2/1) to obtain the preparation of (S)-2-((tert-butoxycarbonyl)amino)propyl-4-toluenesulfonate (I-15a, 6.8 g, yield 72.34%).

Step 2: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (I-15b)

At room temperature, cesium carbonate (I-15a, 16.81 g, 51.60 mmol) and (S)-2-((tert-butoxycarbonyl)amino)propyl-4-toluenesulfonate (6.8 g, 20.64 mmol) were added to a solution of 1H-pyrrole-3-carboxylic acid methyl ester (3.1 g, 17.20 mmol) in N,N-dimethylformamide (80 ml), and the resulting solution was stirred at 80°C overnight. Under ice bath, the reaction solution was quenched with water (100 mL), stirred for 30 minutes and then filtered. The filter cake was dried at 80°C for 4 hours to obtain (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (I-15b, 2.71 g, yield 55.8%). ESI[M+H] ⁺ =283.3

Step 3: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (I-15c)

At room temperature, sodium hydroxide (666 mg, 16.65 mmol) and water (5 mL) were added to a solution of (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (I-15b, 2.35 g, 8.32 mmol) in methanol (30 mL), and the reaction solution was reacted overnight at 80°C in a vacuum tube. The reaction solution was subjected to reduced pressure distillation to remove the solvent, and water (20 mL) was added. The pH was adjusted to 2 with 4M hydrochloric acid solution, and (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (I-15c, 2.2 g, yield 98.51%) was obtained by filtration. ESI[M+H] ⁺ =269.2

Step 4: Preparation of (S)-1-(2-aminopropyl)-1H-pyrrole-3-carboxylic acid (I-15d)

At room temperature, a 4M solution of dioxane hydrochloride (15 mL) was added to a solution of (2-((tert-butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (I-15c, 2.2 g, 8.2 mmol) in ethyl acetate (15 mL). The reaction solution was reacted at room temperature for 3 hours and then filtered to obtain (S)-1-(2-aminopropyl)-1H-pyrrole-3-carboxylic acid (I-15d, 1.35 g, yield 98.7%). ESI[M+H] ⁺ =169.2

Step 5: Preparation of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (I-15)

To a solution of (S)-1-(2-aminopropyl)-1H-pyrrole-3-carboxylic acid (I-15d, 500 mg, 2.96 mmol) in ethanol (15 mL) were added triethylamine (1.24 mL, 8.88 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (1.07 g, 3.25 mmol) at room temperature. ), stirred at 60°C for 4 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (dichloromethane: methanol = 10: 1) to obtain (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (I-15, 920 mg, yield 56.0%). ESI[M-100+H]+＝361.2

(16) Intermediate I-16: Preparation of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (I-16)

At room temperature, (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (I-15, 510 mg, 1.107 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (20 mL), and the reaction solution was stirred at room temperature overnight. After reduced pressure distillation, the crude product was purified by silica gel chromatography (dichloromethane: methanol = 10: 1) to obtain (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (I-16, 90.2 mg, yield 25.15%). ESI[M+H] ⁺ = 331.6

(17) Intermediate I-17: Preparation of (S)-2-(3-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-17)

Step 1: Preparation of (S)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-17a)

Potassium carbonate (275 mg, 1.998 mmol) and 2-chloro-5-(trifluoromethyl)pyrimidine (182 mg, 0.999 mmol) were added to a solution of (S)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.999 mmol) in N-methylpyrrolidone (10 ml) at room temperature, and the reaction solution was stirred at 80°C overnight. 20 mL of water was added to quench the reaction, and 3×10 mL of ethyl acetate were extracted. The organic layers were combined, washed with 3×10 mL of water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. (S)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-17a, 286 mg, yield 82.7%) was obtained. ESI[M+H] ⁺ =347.3

Step 2: Preparation of (S)-2-(3-methylpiperazine-1-yl)-5-(trifluoromethyl)pyrimidine (I-17)

At room temperature, 4M hydrochloric acid dioxane solution (3 mL) was added to a solution (3 mL) of ethyl acetate (S)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-17a, 286 mg, 0.826 mmol). The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to obtain (S)-2-(3-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-17, 120 mg, yield 59.02%). ESI[M+H] ⁺ = 247.2

(18) Intermediate I-18: Preparation of (R)-2-(3-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-18)

Step 1: Preparation of (R)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-18a)

Potassium carbonate (138 mg, 0.999 mmol) and 2-chloro-5-(trifluoromethyl)pyrimidine (91 mg, 0.499 mmol) were added to a solution of (R)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (100 mg, 0.499 mmol) in N-methylpyrrolidone (10 ml) at room temperature, and the reaction solution was stirred at 80°C overnight. 20 mL of water was added to quench the reaction, and 3×10 mL of ethyl acetate were extracted. The organic layers were combined, washed with 3×10 mL of water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. (R)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-18a, 110 mg, yield 63.61%) was obtained. ESI[M+H] ⁺ =347.3

Step 2: Preparation of (R)-2-(3-methylpiperazine-1-yl)-5-(trifluoromethyl)pyrimidine (I-18)

At room temperature, 4M hydrochloric acid dioxane solution (3 mL) was added to a solution (3 mL) of ethyl acetate (R)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-18a, 110 mg, 0.318 mmol). The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to obtain (R)-2-(3-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-18, 100 mg, crude product). ESI[M+H] ⁺ =247.2

(19) Intermediate I-19: Preparation of 2-(piperidin-4-yl)-5-(trifluoromethyl)pyrimidine (I-19)

Step 1: Preparation of tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (I-19a)

To a solution of tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxybenzaldehyde-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (200 mg, 0.647 mmol) in 1,4-dioxane at room temperature were added 2-chloro-5-(trifluoromethyl)pyrimidine (118 mg, 0.647 mmol), potassium carbonate (268.2 mg, 2.022 mmol), [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride (23.7 mg, 0.024 mmol) and water (0.2 ml), and the reaction solution was stirred at 90 degrees Celsius overnight under nitrogen protection. After the reaction solution was filtered, the filter cake was washed with ethyl acetate, and the filtrate was concentrated under reduced pressure to obtain tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (I-19a, 210 mg, yield 98.59%). ESI[M-54+H] ⁺ =274.0

Step 2: Preparation of tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidine-1-carboxylate (I-19b)

At room temperature, 5% palladium on carbon (20 mg) was added to a methanol solution (10 mL) of tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (I-19a, 100 mg, 0.304 mmol), and the mixture was stirred overnight at room temperature under a hydrogen atmosphere. After filtering the reaction solution, the filter cake was washed with methanol. After the filtrate was concentrated under reduced pressure, the crude product was purified by silica gel chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidine-1-carboxylate (I-19b, 32 mg, yield 31.81%). ESI[M+H] ⁺ = 332.2

Step 3: Preparation of 2-(piperidin-4-yl)-5-(trifluoromethyl)pyrimidine (I-19)

At room temperature, 4M hydrochloric acid dioxane solution (3 mL) was added to a solution (3 mL) of 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidine-1-carboxylic acid tert-butyl ester (I-19b, 32 mg, 0.097 mmol) in ethyl acetate. The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to obtain 2-(piperidin-4-yl)-5-(trifluoromethyl)pyrimidine (I-19, 28 mg, crude product). ESI[M+H] ⁺ =322.2

(20) Intermediate I-20: Preparation of 2-(piperazin-1-yl)-4-(trifluoromethyl)pyrimidine (I-20)

Step 1: Preparation of tert-butyl 4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-20a)

Potassium carbonate (296.7 mg, 2.148 mmol) and 2-chloro-4-(trifluoromethyl)pyrimidine (196 mg, 1.074 mmol) were added to a solution of tert-butyl piperazine-1-carboxylate (200 mg, 1.074 mmol) in N-methylpyrrolidone (10 ml) at room temperature, and the reaction solution was stirred at 80°C overnight. Under ice bath, 20 mL of water was added to quench the reaction, and the filter cake was washed with water after filtration. The filter cake was dried at 80°C for 12 hours to obtain tert-butyl 4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I acid 20a, 330 mg, yield 92.48%). ESI [M+H] ⁺ = 333.2

Step 2: Preparation of 2-(piperazin-1-yl)-4-(trifluoromethyl)pyrimidine (I-20)

At room temperature, 4M hydrochloric acid dioxane solution (5 mL) was added to a solution (5 mL) of 4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-20a, 330 mg, 0.993 mmol) in ethyl acetate. The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to obtain 2-(piperazine-1-yl)-4-(trifluoromethyl)pyrimidine (I-20, 180 mg, yield 78.06%). ESI[M+H] ⁺ = 233.2

(21) Intermediate I-21: Preparation of 2-(piperazin-1-yl)pyrimidine-4-carbonitrile (I-21)

Step 1: Preparation of tert-butyl 4-(4-cyanopyrimidin-2-yl)piperazine-1-carboxylate (I-21a)

Potassium carbonate (296.7 mg, 2.148 mmol) and 2-chloropyrimidine-4-carbonitrile (150 mg, 1.074 mmol) were added to a solution of tert-butyl piperazine-1-carboxylate (200 mg, 1.074 mmol) in N-methylpyrrolidone (10 ml) at room temperature, and the reaction solution was stirred at 80°C overnight. Under ice bath, 20 mL of water was added to quench the reaction, and the filter cake was washed with water after filtration. The filter cake was dried at 80°C for 12 hours to obtain tert-butyl 4-(4-cyanopyrimidin-2-yl)piperazine-1-carboxylate (I-21a, 246 mg, yield 79.18%). ESI [M + H] ⁺ = 290.3

Step 2: Preparation of 2-(piperazine-1-yl)pyrimidine-4-carbonitrile (I-21)

At room temperature, 4M hydrochloric acid dioxane solution (5 mL) was added to a solution (5 mL) of 4-(4-cyanopyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-21a, 246 mg, 0.85 mmol) in ethyl acetate. The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to obtain 2-(piperazine-1-yl)pyrimidine-4-carbonitrile (I-21, 100 mg, yield 62.16%). ESI [M + H] ⁺ = 190.2

(22) Intermediate I-22: Preparation of (S)-2-(3-isopropylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-22)

Step 1: Preparation of (S)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-22a)

Potassium carbonate (60.4 mg, 0.876 mmol) and 2-chloro-5-(trifluoromethyl)pyrimidine (80 mg, 0.438 mmol) were added to a solution of (S)-2-isopropylpiperazine-1-carboxylic acid tert-butyl ester (100 mg, 0.348 mmol) in N-methylpyrrolidone (10 ml) at room temperature, and the reaction solution was stirred at 80°C overnight. 20 mL of water was added to quench the reaction, and 3×10 mL of ethyl acetate were extracted. The organic layers were combined, washed with 3×10 mL of water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. (S)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-22a, 132 mg, yield 80.5%) was obtained. ESI[M+H] ⁺ =375.2

Step 2: Preparation of (S)-2-(3-isopropylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-22)

At room temperature, 4M hydrochloric acid dioxane solution (5 mL) was added to a solution (5 mL) of (S)-2-(3-isopropylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-22a, 132 mg, 0.353 mmol) in ethyl acetate. The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to obtain (S)-2-(3-isopropylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-22, 100 mg, crude product). ESI[M+H] ⁺ =274.2

(23) Intermediate I-23: Preparation of (R)-3-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine (I-23)

Step 1: Preparation of (R)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-23a)

Potassium carbonate (276 mg, 1.998 mmol) and 2-chloro-5-(trifluoromethyl)pyridine (180.9 mg, 0.999 mmol) were added to a solution of (R)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.999 mmol) in N-methylpyrrolidone (10 ml) at room temperature, and the reaction solution was stirred overnight at 80 degrees. 20 mL of water was added to quench the reaction, and 3×10 mL of ethyl acetate were extracted. The organic layers were combined, washed with 3×10 mL of water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. (R)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-23a, 210 mg, yield 60.89%) was obtained. ESI[M+H] ⁺ =346.3

Step 2: Preparation of (R)-3-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine (I-23)

At room temperature, 4M hydrochloric acid dioxane solution (5 mL) was added to a solution (5 mL) of (R)-3-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine (I-23a, 210 mg, 0.608 mmol) in ethyl acetate. The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to obtain (R)-3-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine (I-23, 101 mg, yield 67.7%). ESI[M+H] ⁺ =246.3

(24) Intermediate 1-24: Preparation of (S)-3-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine (I-24)

Step 1: Preparation of (S)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-24a)

Potassium carbonate (276 mg, 1.998 mmol) and 2-chloro-5-(trifluoromethyl)pyridine (180.9 mg, 0.999 mmol) were added to a solution of (S)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.999 mmol) in N-methylpyrrolidone (10 ml) at room temperature, and the reaction solution was stirred at 80°C overnight. 20 mL of water was added to quench the reaction, and 3×10 mL of ethyl acetate were extracted. The organic layers were combined, washed with 3×10 mL of water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. (S)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-24a, 210 mg, yield 60.89%) was obtained. ESI[M+H] ⁺ =346.3

Step 2: Preparation of (S)-3-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine (I-24)

At room temperature, 4M hydrochloric acid dioxane solution (5 mL) was added to a solution (5 mL) of (S)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-24a, 210 mg, 0.608 mmol) in ethyl acetate. The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to obtain (R)-3-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine (I-24, 100 mg, yield 67.1%). ESI[M+H] ⁺ =246.3

(25) Intermediate I-25: Preparation of (R)-2-(3-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrazine (I-25)

Step 1: Preparation of (R)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-25a)

Potassium carbonate (151 mg, 1.098 mmol) and 2-chloro-5-(trifluoromethyl)pyrazine (100 mg, 0.549 mmol) were added to a solution of (R)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (110 mg, 0.549 mmol) in N-methylpyrrolidone (10 ml) at room temperature, and the reaction solution was stirred at 80°C overnight. 20 mL of water was added to quench the reaction, and 3×10 mL of ethyl acetate were extracted. The organic layers were combined, washed with 3×10 mL of water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. (R)-2-methyl-4-(5-(trifluoromethyl)pyrazine-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-25a, 120 mg, yield 63.08%) was obtained. ESI[M+H] ⁺ =347.2

Step 2: Preparation of (R)-2-(3-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrazine (I-25)

At room temperature, 4M hydrochloric acid dioxane solution (5 mL) was added to a solution (5 mL) of ethyl acetate (R)-2-methyl-4-(5-(trifluoromethyl)pyrazine-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-25a, 120 mg, 0.302 mmol). The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to obtain (R)-2-(3-methylpiperazine-1-yl)-5-(trifluoromethyl)pyrazine (I-25, 52 mg, yield 60.95%). ESI[M+H] ⁺ = 247.2

(26) Intermediate I-26: Preparation of 2-(piperazin-1-yl)pyrimidine-5-carbonitrile (I-26)

Step 1: Preparation of tert-butyl 4-(5-fluoropyrimidin-2-yl)piperazine-1-carboxylate (I-26a)

6-Chloropyrimidine-5-carbonitrile (75 mg, 1 eq.) was dissolved in N-methylpyrrolidone (4 ml), and piperazine-1-carboxylic acid tert-butyl ester (100 mg, 1 eq.) and potassium carbonate (148 mg, 2 eq.) were added, and the mixture was reacted at 80°C for 1 hour. Water (100 ml) was added, and the solid was collected by filtration and dried in vacuo to obtain the target product 4-(5-cyanopyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-26a, 190 mg, purity 80%, yield 98.18%).

Step 2: Preparation of 2-(piperazine-1-yl)pyrimidine-5-carbonitrile (I-26)

Dissolve tert-butyl 4-(5-cyanopyrimidin-2-yl)piperazine-1-carboxylate (I-26a, 190 mg, purity 80%, 1 eq.) in dichloromethane (2 ml), add trifluoroacetic acid (2 ml) dropwise under ice bath, and react at room temperature for 1 hour. Concentrate the solid under reduced pressure to obtain the target product 2-(piperazin-1-yl)pyrimidine-5-carbonitrile (I-26, 120 mg, purity 80%, yield 96.75%).

(27) Intermediate I-27: Preparation of (R)-2-(2-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-27)

Step 1: Preparation of (R)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-27a)

2-Chloro-5-(trifluoromethyl)pyrimidine (110 mg, 1.2 eq.) was dissolved in N-methylpyrrolidone (5 ml), and tert-butyl (R)-3-methylpiperazine-1-carboxylate (100 mg, 1 eq.) and potassium carbonate (207 mg, 3 eq.) were added, and the mixture was reacted at 80°C for 1 hour. Saturated brine (20 ml) was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the target product (R)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-27a, 139 mg, yield 80.38%). ESI[M-56+H] ⁺ =291.1, ESI[M-100+H] ⁺ =247.0

Step 2: Preparation of (R)-2-(2-methylpiperazine-1-yl)-5-(trifluoromethyl)pyrimidine (I-27)

(R)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-27a, 139 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (4 ml) and reacted at room temperature for 1 hour. The reaction was concentrated under reduced pressure to obtain the target product (R)-2-(2-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-27, 160 mg, purity 50%, yield 80.95%).

(28) Intermediate I-28: Preparation of 2-((3R,5S)-3,5-dimethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-28)

Step 1: Preparation of tert-butyl (2R, 6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-28a)

Dissolve (2R,6S)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (100 mg, 1 eq.) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyrimidine (85 mg, 1 eq.) and potassium carbonate (193 mg, 3 eq.), and react at 80°C for 1 hour. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product (2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-28a, 110 mg, yield 65.4%).

Step 2: Preparation of 2-((3R,5S)-3,5-dimethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-28)

Dissolve (2R, 6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-28a, 110 mg, 1 eq.) in 4M hydrochloric acid diethyl ester (4 ml) and react at room temperature for 1 hour. Collect the precipitated solid by filtration and wash with ethyl acetate to obtain the target product 2-((3R, 5S)-3,5-dimethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-28, 70 mg, yield 88.1%). ESI[M+H] ⁺ 261.1

(29) Intermediate I-29: Preparation of 2-(piperazin-1-yl)-5-(trifluoromethyl)pyrazine (I-29)

Step 1: Preparation of tert-butyl 4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carboxylate (I-29a)

Dissolve tert-butyl piperazine-1-carboxylate (100 mg, 1 eq.) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyrazine (98 mg, 1 eq.) and potassium carbonate (223 mg, 3 eq.), and react at 80°C for 1 hour. Add water (50 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product tert-butyl 4-(5-(trifluoromethyl)pyrazine-2-yl)piperazine-1-carboxylate (I-29a, 140 mg, yield 78.46%). ESI[M+H] ⁺ =333.2

Step 2: Preparation of 2-(piperazin-1-yl)-5-(trifluoromethyl)pyrazine (I-29)

Dissolve tert-butyl 4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carboxylate (I-29a, 140 mg, 1 eq.) in 4M hydrochloric acid ethyl acetate solution (4 ml) and react at room temperature for 1 hour. Collect the precipitated solid by filtration and wash with ethyl acetate to obtain the target product 2-(piperazin-1-yl)-5-(trifluoromethyl)pyrazine (I-29, 72 mg, yield 73.60%). ESI[M+H] ⁺ =233.1

(30) Intermediate I-30: Preparation of 6-chloro-3-((2-(trimethylsilyl)ethoxy)methyl)pyrimidin-4(3H)-one (I-30)

6-Chloropyrimidin-4(3H)-one (100 mg, 1 eq.) was dissolved in N,N-dimethylformamide (4 ml), and 60% sodium hydroxide (46 mg, 1.5 eq.) was added under nitrogen protection in an ice bath, and stirred at room temperature for 30 minutes. (2-(chloromethoxy)ethyl)trimethylsilane (192 mg, 1.5 eq.) was added in an ice bath, and stirred at room temperature for 5 hours. Water (60 ml) was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a flash column (petroleum ether: ethyl acetate = 4:1) to obtain the target product 6-chloro-3-((2-(trimethylsilyl)ethoxy)methyl)pyrimidin-4(3H)-one (I-30, 70 mg, yield 35.04%).

(31) Intermediate I-31: Preparation of (R)-2-(3-ethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-31)

Step 1: Preparation of (R)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-31a)

Dissolve (R)-2-ethylpiperazine-1-carboxylic acid tert-butyl ester (100 mg, 1 eq.) in N-methylpyrrolidone (5 ml), add 2-chloro-5-(trifluoromethyl)pyrimidine (102 mg, 1.2 eq.) and potassium carbonate (193 mg, 3 eq.), and react at 80°C for 1 hour. Add water (30 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product (R)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-31a, 170 mg, purity 80%, yield 79.8%).

Step 2: Preparation of (R)-2-(3-ethylpiperazine-1-yl)-5-(trifluoromethyl)pyrimidine (I-31)

Dissolve (R)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-31a, 170 mg, purity 80%, 1 eq.) in 4M hydrochloric acid ethyl acetate solution (4 ml) and react at room temperature for 1 hour. Collect the precipitated solid by filtration and wash with ethyl acetate to obtain the target product (R)-2-(3-ethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-31, 80 mg, yield 81.45%). ESI[M+H] ⁺ =261.1

(32) Intermediate I-32: Preparation of (R)-2-(3-isopropylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-32)

Step 1: Preparation of (R)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-32a)

Tert-butyl (R)-2-isopropylpiperazine-1-carboxylate (100 mg, 1 eq.) was dissolved in N-methylpyrrolidone (4 ml), and 2-chloro-5-(trifluoromethyl)pyridine (96 mg, 1.2 eq.) and potassium carbonate (182 mg, 3 eq.) were added, and the mixture was reacted at 80°C for 1 hour. Water (20 ml) was added, and the mixture was stirred at 0°C for 30 minutes. The precipitated solid was collected by filtration, washed with water, and dried at 70°C to obtain the target product, tert-butyl (R)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-32a, 150 mg, yield 91.48%).

Step 2: Preparation of (R)-2-(3-isopropylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-32)

Dissolve (R)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-32a, 150 mg, 1 eq.) in 4M ethyl dioxyhydrochloride (4 ml) and react at room temperature for 1 hour. Collect the precipitated solid by filtration and wash with ethyl acetate to obtain the target product (R)-2-(3-isopropylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-32, 76 mg, yield 69.16%). ESI[M+H] ⁺ =275.2

(33) Intermediate I-33: Preparation of (S)-2-(3-ethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-33)

Step 1: Preparation of (S)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-33a)

Dissolve (S)-2-ethylpiperazine-1-carboxylic acid tert-butyl ester (100 mg, 1 eq.) in N-methylpyrrolidone (4 ml), add 2-chloro-5-(trifluoromethyl)pyrimidine (102 mg, 1.2 eq.) and potassium carbonate (193 mg, 3 eq.), and react at 80°C for 1 hour. Add water (20 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product (S)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-33a, 180 mg, purity 80%, yield 96.34%).

Step 2: Preparation of (S)-2-(3-ethylpiperazine-1-yl)-5-(trifluoromethyl)pyrimidine (I-33)

Dissolve (S)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-33a, 180 mg, purity 80%, 1 eq.) in 4M ethyl dioxyhydrochloride (3 ml) and react at room temperature for 1 hour. Collect the precipitated solid by filtration and wash with ethyl acetate to obtain the target product (S)-2-(3-ethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-33, 60 mg, yield 51.28%). ESI[M+H] ⁺ =261.1

(34) Intermediate I-34: Preparation of (S)-2-(3-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrazine (I-34)

Step 1: Preparation of (S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-34a)

Dissolve (S)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (100 mg, 1 eq.) in N-methylpyrrolidone (4 ml), add 2-chloro-5-(trifluoromethyl)pyrazine (109 mg, 1.2 eq.) and potassium carbonate (207 mg, 3 eq.), and react at 80°C for 4 hours. Add water (10 ml), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry at 70°C to obtain the target product (S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-34a, 190 mg, purity 80%, yield 87.9%).

Step 2: Preparation of (S)-2-(3-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrazine (I-34)

Dissolve (S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-34a, 190 mg, purity 80%, 1 eq.) in 4M ethyl dioxyhydrochloride (3 ml) and react at room temperature for 1 hour. Collect the precipitated solid by filtration and wash with ethyl acetate to obtain the target product (S)-2-(3-ethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (I-34, 115 mg, yield 85.14%). ESI[M+H] ⁺ =247.0

(35) Intermediate I-35: Preparation of 1-(4-(trifluoromethyl)cyclohexyl)piperazine (I-35)

Step 1: Preparation of tert-butyl 4-(4-(trifluoromethyl)cyclohexyl)piperazine-1-carboxylate (I-35a)

4-(Trifluoromethyl)cyclohexane-1-one (100 mg, 1 eq.), tert-butyl piperazine-1-carboxylate (90 mg, 0.8 eq.) were dissolved in dichloromethane (3 ml), sodium triacetoxyborohydride (99 mg, 1 eq.) and acetic acid (18 mg, 0.5 eq.) were added, and the mixture was reacted at room temperature overnight. 1N sodium hydroxide solution (1 ml) was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a Flash column (ethyl acetate: petroleum ether = 5% to 20%) to obtain the target product tert-butyl 4-(4-(trifluoromethyl)cyclohexyl)piperazine-1-carboxylate (I-35a, 47 mg, yield 23.21%).

Step 2: Preparation of 1-(4-(trifluoromethyl)cyclohexyl)piperazine (I-35)

Dissolve tert-butyl 4-(4-(trifluoromethyl)cyclohexyl)piperazine-1-carboxylate (I-35a, 47 mg, 1 eq.) in 4M ethyl dioxyhydrochloride (2 ml) and react at room temperature for 1 hour. Collect the precipitated solid by filtration and wash with ethyl acetate to obtain the target product 1-(4-(trifluoromethyl)cyclohexyl)piperazine (I-35, 20 mg, yield 60.58%). ESI[M+H] ⁺ =237.0

(36) Preparation of Intermediate I-36: 1-(4-(trifluoromethyl)phenyl)piperazine (I-36)

Step 1: Preparation of tert-butyl 4-(4-(trifluoromethyl)phenyl)piperazine-1-carboxylate (I-36a)

Piperazine-1-carboxylic acid tert-butyl ester (100 mg, 1 eq.) was dissolved in 1.4-dioxane (6 ml), 1-bromo-4-(trifluoromethyl)benzene (121 mg, 1 eq.), cesium carbonate (350 mg, 2 eq.) were added, and 2,2′-bis(diphenylphosphinoyl)-1,1′-binaphthyl (20 mg. 0.06 eq.) and diacetoxypalladium (5 mg, 0.04 eq.) were added under nitrogen protection, and the mixture was reacted at 85° C. overnight. Water (20 ml) was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified with a Flash column (ethyl acetate: petroleum ether = 5% to 20%) to obtain the target product 4-(4-(trifluoromethyl)phenyl)piperazine-1-carboxylic acid tert-butyl ester (I-36a, 72 mg, yield 40.59%). ESI[M+H] ⁺ ＝331.2

Step 2: Preparation of 1-(4-(trifluoromethyl)phenyl)piperazine (I-36)

Dissolve tert-butyl 4-(4-(trifluoromethyl)phenyl)piperazine-1-carboxylate (I-36a, 72 mg, 1 eq.) in 4M ethyl dioxyhydrochloride (2 ml) and react at room temperature for 1 hour. Collect the precipitated solid by filtration and wash with ethyl acetate to obtain the target product 1-(4-(trifluoromethyl)phenyl)piperazine (I-36, 80 mg, purity 50%, yield 79.71%). ESI[M+H] ⁺ =231.0

(37) Intermediate 1-37: Preparation of 2-(piperazin-1-yl-2,2,3,3,5,5,6,6-d8)-5-(trifluoromethyl)pyrimidine hydrochloride (I-37)

Step 1: Preparation of tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid-2,2,3,3,5,5,6,6-d8 (I-37a)

2-Chloro-5-(trifluoromethyl)pyrimidine (56 mg, 0.308 mmol) was dissolved in N-methylpyrrolidone (4 mL), tert-butylpiperazine-1-carboxylic acid-2,2,3,3,5,5,6,6-d8 (50 mg, 0.257 mmol) and potassium carbonate (107 mg, 0.771 mmol) were added, and the mixture was reacted at 80°C for 2 hours. Water (10 mL) was added, the solid was collected by filtration, and vacuum dried to obtain the target product tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid-2,2,3,3,5,5,6,6-d8 (I-37a) (87 mg, yield 99.33%). ESI[M+H-56] ⁺ = 285.0, ESI[M+H-100] ⁺ : 241.1

Step 2: Preparation of 2-(piperazine-1-yl-2,2,3,3,5,5,6,6-d8)-5-(trifluoromethyl)pyrimidine hydrochloride (I-37)

Tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid-2,2,3,3,5,5,6,6-d8 (I-37a) (87 mg, 0.255 mmol) was dissolved in 4M hydrochloric acid ethyl acetate (2 mL) and reacted at room temperature for 1 hour. The reaction solution was filtered and the solid was washed with petroleum ether to obtain the target product 2-(piperazine-1-yl-2,2,3,3,5,5,6,6-d8)-5-(trifluoromethyl)pyrimidine hydrochloride (I-37) (63 mg, yield 89.08%). ESI[M+H] ⁺ =241.0

(38) Intermediate I-38: Preparation of 1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-38)

Step 1: Preparation of tert-butyl 4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (I-38a)

Dissolve 2-chloro-5-(trifluoromethyl)pyridine (500 mg, 2.74 mmol) in N-methylpyrrolidone (20 mL), add piperazine-1-carboxylic acid tert-butyl ester (510 mg, 2.74 mmol) and potassium carbonate (1.13 g, 8.22 mmol), and react at 80°C for 2 hours. Add saturated brine (20 mL) to quench, extract with ethyl acetate, wash the organic phase with saturated brine, dry over anhydrous magnesium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain the target product 4-(5-methylpyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-38a, 0.37 g, yield 40.54%). ESI[M+H] ⁺ =332.2

Step 2: Preparation of 1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-38)

4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-38a, 0.37 g, 1.12 mmol) was added to 4M hydrochloric acid dioxane solution (5 mL) and reacted at room temperature for 2 hours. The precipitated solid was collected by filtration and washed with ethyl acetate to obtain the target product 1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-38, 320 mg, purity 80%, yield 85.64%). ESI[M+H] ⁺ =232.0

(39) Intermediate I-39: Preparation of 1-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-39)

Step 1: Preparation of tert-butyl 4-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (I-39a)

Dissolve tert-butyl piperazine-1-carboxylate (114 mg, 0.612 mmol) in N-methylpyrrolidone (5 mL), add 2-bromo-3-fluoro-5-(trifluoromethyl)pyridine (150 mg, 0.612 mmol) and potassium carbonate (253 mg, 1.836 mmol), and react at 90°C for 3 hours. Add water (20 mL), stir at 0°C for 30 minutes, collect the precipitated solid by filtration, wash with water, and dry to obtain the target product tert-butyl 4-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (I-39a, 200 mg, yield 93.54%). ESI[M-56+H] ⁺ =294.1, ESI[M-100+H] ⁺ =249.9

Step 2: Preparation of 1-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-39)

Dissolve tert-butyl 4-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (I-39a, 200 mg, 0.573 mmol) in 4M hydrochloric acid ethyl acetate solution (4 mL) and react at room temperature for 2 hours. Collect the precipitated solid by filtration and wash with ethyl acetate to obtain the target product 1-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (I-39, 100 mg, yield 61.14%). ESI[M+H] ⁺ =250.0

(40) Intermediate I-40: Preparation of 3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane hydrochloride (I-40)

Step 1: Preparation of tert-butyl 3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (I-40a)

2-Chloro-5-trifluoromethylpyridine (257 mg, 1.42 mmol) was dissolved in N-methylpyrrolidone (10 mL), 8-Boc-3,8-diazabicyclo[3.2.1]octane (300 mg, 1.42 mmol) and potassium carbonate (391 mg, 2.84 mmol) were added, and the mixture was reacted at 100°C for 3 hours. Water (50 mL) was added, the solid was collected by filtration, and vacuum dried to obtain the target product, tert-butyl 3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (I-40a, 300 mg, yield 59.4%).

Step 2: Preparation of 3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane hydrochloride (I-40)

3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (I-40a, 300 mg, 0.84 mmol) was dissolved in 4M hydrochloric acid ethyl acetate (10 mL) and reacted at room temperature for 2 hours. The solid was collected by filtration and washed with petroleum ether to obtain the target product 3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane hydrochloride (I-40, 224 mg, yield 91.1%).

(41) Intermediate I-41: Preparation of 3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane hydrochloride (I-41)

Step 1: Preparation of tert-butyl 3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (I-41a)

2-Chloro-5-(trifluoromethyl)pyrazine (200 mg, 1.1 mmol) was dissolved in N-methylpyrrolidone (10 mL), 8-Boc-3,8-diazabicyclo[3.2.1]octane (233 mg, 1.1 mmol) and potassium carbonate (303 mg, 2.2 mmol) were added, and the mixture was reacted at 100°C for 3 hours. Water (50 mL) was added, the solid was collected by filtration, and vacuum dried to obtain the target product, tert-butyl 3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (I-41a, 354 mg, yield 90.3%). ESI[M+H] ⁺ =359.2, ESI[M-56+H] ⁺ =303.1

Step 2: Preparation of 3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane hydrochloride (I-41)

3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (354 mg, 0.99 mmol) was dissolved in 4M hydrochloric acid ethyl acetate (10 mL) and reacted at room temperature for 3 hours. The solid was collected by filtration and washed with petroleum ether to obtain the target product 3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane hydrochloride (I-41, 290 mg, yield 99.6%).

(42) Intermediate I-42: Synthesis of 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-42)

Step 1: Preparation of 4,5-dibromo-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-42a)

At 0°C, under nitrogen protection, sodium hydride (9.45 g, 236 mol, purity 60%) was added in batches to a stirred solution of 4,5-dibromopyridin-3(2H)-one (40 g, 157.55 mmol) in N,N-dimethylformamide (250 mL). The obtained solution was stirred at room temperature for 0.5 h, then the reaction solution was cooled to 0°C, and (2-(chloromethoxy)ethyl)trimethylsilane (28.89 g, 173.31 mmol) was added dropwise. After the reaction mixture was warmed to room temperature and stirred for 2 h, 250 mL of water was added to quench. The resulting reaction mixture was extracted with ethyl acetate three times, 250 mL each time, and the combined organic extracts were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 4,5-dibromo-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-42a) (63.38 g). ESI[M+Na] ⁺ =407.0, ESI[2M+Na] ⁺ =791.0

Step 2: Preparation of 4-bromo-5-chloro-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-42b)

Under stirring, lithium chloride (6.99 g, 164.99 mol) was added to a solution of 4,5-dibromo-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazine-3(2H)-one (I-42a) (63.38 g, 164.99 mmol) in N-methylpyrrolidone (250 mL). The resulting solution was heated to 95 ° C and stirred for 4 hours, then the reaction solution was cooled to room temperature and quenched by adding 250 mL of water. The resulting reaction mixture was extracted with 3×250 mL of ethyl acetate, and the organic layers were combined. The organic layer was washed with 3×250 mL of brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography (ethyl acetate: petroleum ether = 1:50) to give compound 4-bromo-5-chloro-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-42b) (38.96 g, yield 69.51%). ESI [2M+Na] ⁺ = 701.1

Step 3: Preparation of 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-42)

To a solution of 4-bromo-5-chloro-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-42b) (38.96 g, 114.69 mmol) in N-methylpyrrolidone (250 mL) at room temperature was added iodide (4.37 g, 22.90 mmol), followed by dropwise addition of compound 2,2-difluoro-2-(fluorosulfonyl)acetic acid methyl ester 66.10 g, 344.07 mmol). The resulting solution was stirred at 80 °C for 2 hours. 250 mL of water was added to quench the reaction, and extracted with 3×250 mL of ethyl acetate. The organic layers were combined and washed with 3×250 mL of brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by flash column (ethyl acetate: petroleum ether = 1: 100) to give compound 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (I-42) (24.01 g, yield 63.67%). ESI [M + Na] ⁺ = 351.1, ESI [2M + Na] ⁺ = 679.0

(43) Intermediate I-43: 2-(piperazin-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-43)

Step 1: Preparation of tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-43a)

2-Chloro-5-(trifluoromethyl)pyrimidine (1825 mg, 10 mmol) and tert-butyl piperazine-1-carboxylate (1862 mg, 10 mmol) were dissolved in N-methylpyrrolidone (30 ml), potassium carbonate (2764 mg, 20 mmol) was added, and the mixture was reacted at 80°C for 1 hour. Water (100 ml) was added, the solid was collected by filtration, and vacuum dried to obtain the target product tert-butyl 4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylate (I-43a) (2.9 g, yield 87%).

Step 2: Preparation of 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-43)

4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (I-43a) (332 mg, 1 mmol) was dissolved in 4M hydrochloric acid ethyl acetate (5 ml) and reacted at room temperature for 2 hours. The reaction solution was filtered and the solid was washed with petroleum ether to obtain the target product 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (I-43) (289 mg, yield 94%). ESI [M+H] ⁺ = 233.1, ¹ HNMR (400 MHz, dmso) δ 9.48 (s, 2H), 8.78 (s, 2H), 4.06 (s, 4H), 3.18 (s, 4H).

(44) Intermediate I-44: (S)-2-((tert-Butyloxycarbonyl)amino)propyl 4-toluenesulfonate (I-44)

Dissolve (S)-tert-butyl (1-hydroxypropan-2-yl)carbamate (1751 mg, 10 mmol) in dichloromethane (10 ml), add 4-dimethylaminopyridine (1833 mg, 15 mmol), cool to 0°C, dissolve p-toluenesulfonyl chloride (2097 mg, 11 mmol) in dichloromethane (10 ml) and add dropwise to the flask, react at room temperature for 2 hours. Wash the organic phase twice with pure water and once with brine, dry the organic phase with anhydrous magnesium sulfate, concentrate the filtrate under reduced pressure, and purify with a Flash column (ethyl acetate: petroleum ether) to obtain the target product (S)-2-((tert-butoxycarbonyl)amino)propyl 4-toluenesulfonate (I-44) (2.1 g, yield 65%). ESI[M+H-56]=274.1, ESI[M+H-100] ⁺ =230.1, ESI[2M+H] ⁺ =659.4

(45) Intermediate I-45: (S)-tert-butyl (1-iodopropane-2-yl)carbamate (I-45)

Take imidazole (1184 mg, 17.4 mmol) and triphenylphosphine (4561 mg, 17.4 mmol) and dissolve them in tetrahydrofuran (25 ml) under nitrogen protection. Control the temperature at 0°C. Add iodine (4.8 g, 18.9 mmol) in batches at this temperature. After adding, stir at 0°C for 30 minutes. Dissolve (S)-tert-butyl (1-hydroxypropan-2-yl)carbamate (2537 mg, 14.5 mmol) in tetrahydrofuran (8 ml) and slowly add it to the above mixture at 0°C. After completion, react at 0°C for 4 hours, and then transfer to room temperature for 6 hours. After the reaction is completed, 60 ml of ethyl acetate is added to the mixture, filtered, and the filtrate is washed with saturated sodium thiosulfate (30 ml*3), and then the organic phase is washed with saturated sodium chloride solution (30 ml*3). Finally, the organic phase is dried over anhydrous sodium sulfate and concentrated. Flash column is used to obtain the target product (S)-(1-iodopropane-2-yl)carbamic acid tert-butyl ester (I-45) (2090 mg, yield 50.6%). ESI[M+H-56] ⁺ =230.0, ESI[2M+H] ⁺ =571.3

Example 1: Preparation of 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-2-yl)acetonitrile (1)

Step 1: Preparation of tert-butyl (S)-3-(cyanomethyl)-4-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazine-1-carboxylate (1-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (25 mg, 0.076 mmol) was dissolved in N,N-dimethylformamide (3 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (43 mg, 0.114 mmol), N,N-diisopropylethylamine (20 μL, 0.114 mmol) and (S)-tert-butyl 3-(cyanomethyl)piperazine-1-carboxylate (17 mg, 0.076 mmol) were added and the reaction was carried out at 80°C overnight. Water (5 ml) was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product, tert-butyl (S)-3-(cyanomethyl)-4-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazine-1-carboxylate (1-a) (12 mg, yield 29.5%). ESI [M+H] ⁺ = 538.4, ESI [2M+H] ⁺ = 1075.5, ESI [M-56+H] ⁺ = 482.3.

Step 2: Preparation of 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-2-yl)acetonitrile (1-b)

Tert-butyl (S)-3-(cyanomethyl)-4-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazine-1-carboxylate (1-a) (12 mg, 0.022 mmol) was dissolved in dichloromethane (2 ml), trifluoroacetic acid (0.3 ml) was added, and the mixture was reacted at room temperature for 2 hours. After concentration under reduced pressure, the target product 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-2-yl)acetonitrile (1-b) (9 mg, yield 91.8%) was obtained. ESI[M+H] ⁺ =438.3, ESI[M+Na] ⁺ =460.2.

Step 3: Preparation of 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-2-yl)acetonitrile (1)

2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-2-yl)acetonitrile (1-b) (9 mg, 0.021 mmol) was dissolved in N-methylpyrrolidone (2 ml), and 2-chloro-5-(trifluoromethyl)pyrimidine (4 mg, 0.021 mmol) and potassium carbonate (6 mg, 0.041 mmol) were added, and the reaction was carried out at 100°C for 2 hours. Water (5 ml) was added, extracted with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-2-yl)acetonitrile (1) (2.12 mg, yield 17.7%). ESI [M+H] ⁺ = 584.4, ESI [2M+H] ⁺ = 1167.5.

Example 2: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[2,3-c]pyridin-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (2)

Step 1: Preparation of (S)-1-(2-(tert-butyloxycarbonyl)amino)propyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxylic acid methyl ester (2-a)

6-Azaindole-3-carboxylic acid methyl ester (200 mg, 1.14 mmol) was dissolved in N, N-dimethylformamide (10 ml), cesium carbonate (1.11 g, 3.41 mmol) was added, and the mixture was reacted at room temperature for 30 minutes, tert-butyl-(1-iodopropane-2-yl)carbamate (421 mg, 1.48 mmol) was added, and the mixture was reacted at 80°C overnight. Water (10 ml) was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a Flash column (dichloromethane: methanol = 0-10%) to obtain the target product (S)-1-(2-(tert-butyloxycarbonyl)amino)propyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxylic acid methyl ester (2-a) (270 mg, yield 71.4%). ESI[M+H] ⁺ =334.2, ESI[2M+H] ⁺ =667.5.

Step 2: Preparation of (S)-1-(2-(tert-butyloxycarbonyl)amino)propyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxylic acid (2-b)

(S)-1-(2-(tert-Butyloxycarbonyl)amino)propyl)-1H-pyrrole[2,3-c]pyridine-3-carboxylic acid methyl ester (2-a) (100 mg, 0.3 mmol) was dissolved in methanol (6 ml), and water (2 ml) and sodium hydroxide (24 mg, 0.6 mmol) were added, and the mixture was reacted at 80° C. in a sealed tube overnight. The pH was adjusted to neutral with 1 equivalent of sodium hydroxide solution, and the residue was dissolved with ethanol after concentration under reduced pressure, and the insoluble matter was filtered out. The filtrate was concentrated under reduced pressure to obtain the target product (S)-1-(2-(tert-Butyloxycarbonyl)amino)propyl)-1H-pyrrole[2,3-c]pyridine-3-carboxylic acid (2-b) (96 mg, yield 100.0%).

Step 3: Preparation of tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[2,3-c]pyridin-1-yl)propan-2-yl)carbamate (2-c)

(S)-1-(2-(tert-Butyloxycarbonyl)amino)propyl)-1H-pyrrolo[2,3-c]pyridine-3-carboxylic acid (2-b) (96 mg, 0.3 mmol) was dissolved in dichloromethane (6 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (172 mg, 0.453 mmol), N,N-diisopropylethylamine (131 μL, 0.75 mmol) and 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (81 mg, 0.3 mmol) were added and reacted at room temperature for 3 hours. Saturated ammonium chloride solution (5 ml) was added, extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 10: 1) to obtain the target product tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[2,3-c]pyridin-1-yl)propane-2-yl)carbamate (2-c) (40 mg, yield 25.0%). ESI [M+H] ⁺ = 534.5

Step 4: Preparation of (S)-(1-(2-aminopropyl)-1H-pyrrolo[2,3-c]pyridin-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (2-d)

Tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[2,3-c]pyridin-1-yl)propane-2-yl)carbamate (2-c) (39 mg, 0.073 mmol) was dissolved in dichloromethane (4 ml), trifluoroacetic acid (0.4 ml) was added, and the mixture was reacted at room temperature for 2 hours. After concentration under reduced pressure, the target product (S)-(1-(2-aminopropyl)-1H-pyrrolo[2,3-c]pyridin-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (2-d) (56 mg, yield 140.0%) was obtained. ESI[M+H] ⁺ =434.3, ESI[2M+H] ⁺ =867.5.

Step 5: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[2,3-c]pyridin-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (2-e)

(S)-(1-(2-Aminopropyl)-1H-pyrrolo[2,3-c]pyridin-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (2-d) (56 mg, 0.102 mmol) was dissolved in ethanol (5 ml), and triethylamine (36 μL, 0.255 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazine-3(2H)-one (33 mg, 0.102 mmol) were added, and the reaction was carried out at 60°C for 4 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 10: 1) to obtain the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[2,3-c]pyridin-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (2-e) (15 mg, yield 20.3%). ESI [M+H] ⁺ = 726.5.

Step 6: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[2,3-c]pyridin-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (2)

(S)-4-(Trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[2,3-c]pyridin-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (2-e) (15 mg, 0.021 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the reaction mixture was purified on a preparative plate (dichloromethane:methanol=10:1) to give the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[2,3-c]pyridin-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (2) (5 mg, yield 40.6%). ESI[M+H] ⁺ =596.4. ¹ H NMR (400MHz, CD ₃ OD) δ 9.11 (s, 1H), 8.64 (s, 2H), 8.29 (d, J = 5.8Hz, 1H), 8.12 (s, 1H), 7.86 (d, J = 5.7Hz, 1H), 7.30 (s, 1H), 4.04 (t, J=5.0Hz, 4H), 3.82 (d, J=5.1Hz, 4H), 3.66 (d, J=2.7Hz, 2H), 2.10-2.00 (m, 1H), 1.31 (s, 3H).

Example 3: Preparation of (S)-5-((1-(5-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (3)

Step 1: Preparation of (5-fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (3-a)

5-Fluoroindole-3-carboxylic acid (100 mg, 0.559 mmol) was dissolved in dichloromethane (6 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (318 mg, 0.838 mmol), N,N-diisopropylethylamine (243 μL, 1.398 mmol) and 2-(piperazin-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (150 mg, 0.559 mmol) were added, and the mixture was reacted at room temperature for 3 hours. The solid was collected by filtration to obtain the target product (5-fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (3-a) (178 mg, yield 81.3%). ESI[M+H] ⁺ =394.2, ESI[2M+H] ⁺ =787.4.

Step 2: Preparation of tert-butyl-(1-(5-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)carbamate (3-b)

(5-Fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (3-a) (178 mg, 0.453 mmol) was dissolved in N,N-dimethylformamide (10 ml), cesium carbonate (113 mg, 1.358 mmol) was added, and the mixture was reacted at room temperature for 30 minutes. Tert-butyl-(1-iodopropane-2-yl)carbamate (168 mg, 0.589 mmol) was added, and the mixture was reacted at 80°C overnight. Water (10 ml) was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product, tert-butyl-(1-(5-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)carbamate (3-b) (80 mg, yield 32.1%). ESI[M+H] ⁺ = 551.4, ESI[2M+H] ⁺ = 1101.6, ESI[M-56+H] ⁺ = 495.3.

Step 3: Preparation of (S)-(1-(2-aminopropyl)-5-fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (3-c)

Tert-butyl-(1-(5-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)carbamate (3-b) (80 mg, 0.145 mmol) was dissolved in dichloromethane (4 ml), trifluoroacetic acid (0.5 ml) was added, and the mixture was reacted at room temperature for 2 hours. After concentration under reduced pressure, the target product (S)-(1-(2-aminopropyl)-5-fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (3-c) (100 mg, yield 121.9%) was obtained.

Step 4: Preparation of (S)-5-((1-(5-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (3-d)

(S)-(1-(2-aminopropyl)-5-fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (3-c) (100 mg, 0.177 mmol) was dissolved in ethanol (5 ml), and triethylamine (62 μL, 0.443 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazine-3(2H)-one (58 mg, 0.177 mmol) were added, and the reaction was carried out at 60°C for 8 hours. After concentration under reduced pressure, the residue was dissolved in dichloromethane, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product (S)-5-((1-(5-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (3-d) (80 mg, yield 60.8%). ESI [M+H] ⁺ = 743.4.

Step 5: Preparation of (S)-5-((1-(5-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (3)

(S)-5-((1-(5-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (3-d) (80 mg, 0.108 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (5 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the reaction mixture was purified on a preparative plate (dichloromethane:methanol=18:1) to give the target product (S)-5-((1-(5-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (3) (45 mg, yield 68.2%). ESI[M+H] ⁺ =613.4. ¹ H NMR (400MHz, CDCl ₃ ) δ 8.44 (s, 2H), 7.47 (s, 1H), 7.28 (s, 1H), 7.19 (dd, J = 9.2, 2.0Hz, 1H), 6.98 (td, J = 8.9, 2.1Hz, 1H), 6.86 (s, 1H), 4 .30 (dd, J=13.8, 2.7Hz, 1H), 4.20 (dd, J=14.7, 9.3Hz, 1H), 4.13 (dd, J=13.8, 8.8Hz, 1H), 3.87 (d, J=3.7Hz, 4H), 3.66 (s, 4H), 1.37 (d, J=6.1Hz, 3H).

Example 4: Preparation of 4-(trifluoromethyl)-5-((1R,2S)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)pyridazin-3(2H)-one (4)

Step 1: Preparation of (1R, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl 4-toluenesulfonate (4-a)

Dissolve (1R, 2R)-2-hydroxycyclohexylcarbamic acid tert-butyl ester (350 mg, 1.63 mmol) in dichloromethane (20 ml), add 4-dimethylaminopyridine (397 mg, 3.25 mmol), triethylamine (452 μL, 3.25 mmol) and p-toluenesulfonyl chloride (465 mg, 2.45 mmol), and react at room temperature for 4 hours. Add saturated ammonium chloride solution (5 ml), extract with dichloromethane, dry with anhydrous magnesium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain the target product (1R, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl 4-toluenesulfonate (4-a) (610 mg, yield 101.6%). ESI[M-56+H] ⁺ =314.2, ESI[M-100+H] ⁺ =270.2, ESI[2M+H] ⁺ =739.5.

Step 2: Preparation of methyl 1-((1S, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylate (4-b)

3-pyrrolyl carboxylic acid methyl ester (121 mg, 0.967 mmol) was dissolved in N, N-dimethylformamide (20 ml), cesium carbonate (4-a) (945 mg, 2.9 mmol) was added, and the mixture was reacted at room temperature for 30 minutes, (1R, 2R)-2-((tert-butoxycarbonyl)amino)cyclohexyl 4-toluenesulfonate (610 mg, 1.652 mmol) was added, and the mixture was reacted at 80°C overnight. The insoluble matter was filtered out, the filtrate was concentrated under reduced pressure, and then dissolved with ethyl acetate, and the insoluble matter was filtered out. The filtrate was concentrated under reduced pressure, and then purified on a Flash column (ethyl acetate: petroleum ether = 10% to 30%) to obtain the target product 1-((1S, 2R)-2-((tert-butoxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid methyl ester (4-b) (210 mg, yield 67.5%). ESI[2M+H] ⁺ =645.5, ESI[M-56+H] ⁺ =267.2.

Step 3: Preparation of 1-((1S, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid (4-c)

1-((1S, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid methyl ester (4-b) (110 mg, 0.341 mmol) was dissolved in methanol (6 ml), and water (2 ml) and sodium hydroxide (28 mg, 0.682 mmol) were added. The mixture was reacted at 80°C in a sealed tube overnight. The pH was adjusted to neutral with 1 equivalent of sodium hydroxide solution, and the residue was dissolved with ethyl acetate after concentration under reduced pressure, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the target product 1-((1S, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid (4-c) (100 mg, yield 95.2%). ESI[2M+H] ⁺ =617.6, ESI[M-56+H] ⁺ =253.1.

Step 4: Preparation of tert-butyl ((1R, 2S)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)carbamate (4-d)

1-((1S,2R)-2-((tert-butoxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid (4-c) (100 mg, 0.276 mmol) was dissolved in dichloromethane (8 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (157 mg, 0.414 mmol), N,N-diisopropylethylamine (120 μL, 0.69 mmol) and 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (74 mg, 0.276 mmol) were added and reacted at room temperature for 5 hours. Saturated ammonium chloride solution (8 ml) was added to quench the reaction, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by Flash column (ethyl acetate: petroleum ether = 20% to 50%) to obtain the target product tert-butyl ((1R, 2S)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)carbamate (4-d) (150 mg, yield 88.8%). ESI [M+H] ⁺ = 523.4, ESI [2M+H] ⁺ = 1045.7, ESI [M-56+H] ⁺ = 467.3.

Step 5: Preparation of (1-((1S,2R)-2-aminocyclohexyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (4-e)

Tert-butyl ((1R, 2S)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)carbamate (4-d) (150 mg, 0.287 mmol) was dissolved in dichloromethane (10 ml), trifluoroacetic acid (1 ml) was added, and the mixture was reacted at room temperature for 2 hours. After concentration under reduced pressure, the target product (1-((1S, 2R)-2-aminocyclohexyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (4-e) (160 mg, yield 103.9%) was obtained.

Step 6: Preparation of 4-(trifluoromethyl)-5-((1R, 2S)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (4-f)

(1-((1S,2R)-2-aminocyclohexyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (4-e) (160 mg, 0.298 mmol) was dissolved in ethanol (10 ml), and triethylamine (104 μL, 0.746 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazine-3(2H)-one (98 mg, 0.298 mmol) were added, and the reaction was carried out at 60°C for 6 hours. After concentration under reduced pressure, the residue was dissolved in dichloromethane, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product 4-(trifluoromethyl)-5-((1R, 2S)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (4-f) (125 mg, yield 58.7%). ESI [M+H] ⁺ = 715.5.

Step 7: Preparation of 4-(trifluoromethyl)-5-((1R, 2S)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)pyridazin-3(2H)-one (4)

4-(Trifluoromethyl)-5-((1R,2S)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (4-f) (125 mg, 0.175 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (8 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the mixture was purified on a preparative plate (dichloromethane:methanol=20:1) to obtain the target product 4-(trifluoromethyl)-5-((1R,2S)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)pyridazin-3(2H)-one (4) (71 mg, yield 69.6%). ESI[M+H] ⁺ =585.3, ESI[2M+H] ⁺ =1169.5. ¹ H NMR (400MHz, CD ₃ OD) δ 8.60 (s, 2H), 7.57 (s, 1H), 7.22 (s, 1H), 6.83 (t, J = 2.5Hz, 1H), 6.28-6.21 (m, 1H), 4.06 (dd, J=15.0, 8.2Hz, 1H), 3.99 (d, J=3.9Hz, 1H), 3.96-3.88 (m, 4H), 3.78-3.66 (m, 4H), 2.07 (m, 3H), 1.91 (m, 2H), 1.58 (m, 3H).

Example 5: Preparation of (S)-5-((1-(3-(4-(5-methylpyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (5)

Step 1: Preparation of (S)-5-((1-(3-(4-(5-methylpyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (5-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (20 mg, 0.043 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (25 mg, 0.065 mmol), N,N-diisopropylethylamine (19 μL, 0.109 mmol) and 5-methyl-2-(piperazin-1-yl)pyrimidine hydrochloride (10 mg, 0.043 mmol) were added and reacted at room temperature for 2 hours. The insoluble matter was filtered, and the filtrate was concentrated under reduced pressure and then purified by preparative plate (dichloromethane: methanol = 17: 1) to obtain the target product (S)-5-((1-(3-(4-(5-methylpyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (5-a) (15 mg, yield 55.6%). ESI [M+H] ⁺ = 621.4.

Step 2: Preparation of (S)-5-((1-(3-(4-(5-methylpyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (5)

(S)-5-((1-(3-(4-(5-methylpyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (5-a) (15 mg, 0.024 mmol) was dissolved in 4M hydrochloric acid dioxane solution (2 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the mixture was purified by preparative plate (dichloromethane: methanol = 18: 1) to obtain the target product (S)-5-((1-(3-(4-(5-methylpyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (5) (3 mg, yield 25.4%). ESI[M+H] ⁺ =491.3, ESI[2M+H] ⁺ =981.6. ¹ H NMR (400MHz, CD ₃ OD) δ8.23 (s, 2H), 7.38 (s, 1H), 7.14 (s, 1H), 6.79-6.72 (m, 1H), 6.31-6.25 (m, 1H), 4.42-4. 31 (m, 1H), 4.20 (dd, J=14.1, 3.7Hz, 1H), 3.96 (dd, J=14.1, 9.9Hz, 1H), 3.76 (dd, J=9.8, 1.9Hz, 8H), 2.15 (s, 3H), 1.35 (d, J=6.5Hz, 3H).

Example 6: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (6)

Step 1: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (6-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (50 mg, 0.109 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (62 mg, 0.163 mmol), N,N-diisopropylethylamine (48 μL, 0.272 mmol) and 3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane hydrochloride (32 mg, 0.109 mmol) were added and reacted at room temperature for 4 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (6-a) (59 mg, yield 77.6%). ESI [M+H] ⁺ = 701.4.

Step 2: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (6)

4-(Trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (6-a) (59 mg, 0.084 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (5 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the mixture was purified by preparative plate (dichloromethane: methanol = 15: 1) to give the target product 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyrimidin-2-yl)-3,8-diazacyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (6) (16.4 mg, yield 34.2%). ESI [M+H] ⁺ = 571.3. ¹ H NMR (400 MHz, CD ₃ OD) δ8.60 (s, 2H), 7.36 (s, 1H), 7.21 (t, J=1.7Hz, 1H), 6.82-6.75 (m, 1H), 6.38 (s, 1H), 4.72-4.58 (m, 4H), 4.37 (dd, J=9.7, 4.5Hz, 1H), 4.22 (dd, J=14. 1, 3.6Hz, 1H), 3.97 (dd, J=14.1, 9.9Hz, 1H), 3.23 (d, J=13.3Hz, 2H), 1.95-1.86 (m, 2H), 1.71 (q, J=5.8Hz, 2H), 1.36 (d, J=6.5Hz, 3H).

Example 7: Preparation of (S)-5-((1-(3-(4-(5-fluoropyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2U)-one (7)

Step 1: Preparation of (S)-5-((1-(3-(4-(5-fluoropyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (7-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (50 mg, 0.109 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (62 mg, 0.163 mmol), N,N-diisopropylethylamine (48 μL, 0.272 mmol) and 5-fluoro-2-(piperazin-1-yl)pyrimidine hydrochloride (24 mg, 0.109 mmol) were added and reacted at room temperature for 4 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product (S)-5-((1-(3-(4-(5-fluoropyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (7-a) (40 mg, yield 59.0%). ESI [M+H] ⁺ = 625.4.

Step 2: Preparation of (S)-5-((1-(3-(4-(5-fluoropyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (7)

(S)-5-((1-(3-(4-(5-fluoropyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (7-a) (40 mg, 0.064 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (5 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the mixture was purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product (S)-5-((1-(3-(4-(5-fluoropyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (7) (17 mg, yield 53.6%). ESI[M+H] ⁺ =495.3, ESI[2M+H] ⁺ =989.5. ¹ H NMR (600MHz, CD ₃ OD) δ8.31 (s, 2H), 7.39 (s, 1H), 7.14 (s, 1H), 6.76 (s, 1H), 6.29 (s, 1H), 4.37 (s, 1H), 4.20 (d, J=11.2Hz, 1H), 3.96 (s, 1H), 3.77 (d, J=34.3Hz, 8H), 1.33 (d, J=34.0Hz, 3H).

Example 8: Preparation of 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (8)

Step 1: Preparation of 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (8-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 0.087 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (50 mg, 0.13 mmol), N,N-diisopropylethylamine (38 μL, 0.217 mmol) and (S)-2-(2-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (25 mg, 0.087 mmol) were added and reacted at room temperature for 2.5 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (8-a) (42 mg, yield 70.2%). ESI [M+H] ⁺ = 689.4.

Step 2: Preparation of 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (8)

5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (8-a) (42 mg, 0.061 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (5 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the mixture was purified by preparative plate (dichloromethane: methanol = 18: 1) to obtain the target product 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (8) (23 mg, yield 67.6%). ESI[M+H] ⁺ =559.4, ESI[2M+H] ⁺ =1117.5. ¹ H NMR (400MHz, CD ₃ OD) δ8.60 (s, 2H), 7.44 (s, 1H), 7.16 (s, 1H), 6.78 (t, J=2.4Hz, 1H), 6.32 (s, 1H), 4.65-4.56 (m, 1H), 4.47-4.24 (m, 3H), 4.21 (dd, J=14.1, 3.7Hz, 1H), 3 .98 (dd, J=14.1, 9.8Hz, 1H), 3.49-3.32 (m, 2H), 3.23 (d, J=8.7Hz, 2H), 1.35 (d, J=6.5Hz, 3H), 1.19 (d, J=6.1Hz, 3H).

Example 9: Preparation of (S)-5-((1-(3-(4-(5-methoxypyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (9)

Step 1: Preparation of (S)-5-((1-(3-(4-(5-methoxypyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (9-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 0.087 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (50 mg, 0.13 mmol), N,N-diisopropylethylamine (38 μL, 0.217 mmol) and 5-methoxy-2-(piperazin-1-yl)pyrimidine hydrochloride (20 mg, 0.087 mmol) were added and reacted at room temperature for 4 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product (S)-5-((1-(3-(4-(5-methoxypyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (9-a) (10 mg, yield 18.2%). ESI [M+H] ⁺ = 637.4.

Step 2: Preparation of (S)-5-((1-(3-(4-(5-methoxypyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (9)

(S)-5-((1-(3-(4-(5-methoxypyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (9-a) (10 mg, 0.016 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (3 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the mixture was purified on a preparative plate (dichloromethane:methanol=18:1) to give the target product (S)-5-((1-(3-(4-(5-methoxypyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (9) (3.2 mg, yield 40.0%). ESI[M+H] ⁺ =507.3.

Example 10: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (10)

Step 1: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (10-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 0.087 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (50 mg, 0.13 mmol), N,N-diisopropylethylamine (38 μL, 0.217 mmol) and 1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (63 mg, 0.236 mmol) were added and reacted at room temperature for 3 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (10-a) (65 mg, yield 40.9%). ESI [M+H] ⁺ = 674.4.

Step 2: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (10)

(S)-4-(Trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (10-a) (65 mg, 0.0965 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (5 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the reaction mixture was purified on a preparative plate (dichloromethane:methanol=15:1) to give the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (10) (10.6 mg, yield 20.2%). ESI[M+H] ⁺ =544.4. ¹ H NMR (400MHz, CD ₃ OD) δ 8.37 (s, 1H), 7.76 (dd, J = 8.9, 1.7Hz, 1H), 7.38 (s, 1H), 7.13 (s, 1H), 6.92 (d, J = 9.0Hz, 1H), 6.78 (s, 1H), 6.30 ( s, 1H), 4.43-4.32 (m, 1H), 4.21 (dd, J=14.1, 3.3Hz, 1H), 3.95 (dd, J=13.9, 10.1Hz, 1H), 3.74 (dd, J=23.7, 4.2Hz, 8H), 1.36 (d, J=6.5Hz, 3H).

Example 11: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (11)

Step 1: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (11-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 0.087 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (50 mg, 0.13 mmol), N,N-diisopropylethylamine (40 μL, 0.226 mmol) and 2-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazacyclo[2.2.1]heptane hydrochloride (27 mg, 0.096 mmol) were added and reacted at room temperature for 4 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (11-a) (46 mg, yield 76.7%). ESI [M+H] ⁺ = 687.4.

Step 2: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (11)

4-(Trifluoromethyl)-5-((2S)-1-(3-(5-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (11-a) (46 mg, 0.067 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (4 ml) and reacted at room temperature for 4 hours. After concentration under reduced pressure, the reaction mixture was purified on a preparative plate (dichloromethane:methanol=15:1) to give the target product, 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(5-(trifluoromethyl)pyrimidin-2-yl)-2,5-diazabicyclo[2.2.1]heptane-2-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (11) (24.8 mg, yield 66.5%). ESI[M+H] ⁺ =557.3. ¹ H NMR (400MHz, CD ₃ OD) δ 8.58 (s, 2H), 7.38 (d, J = 37.8Hz, 1H), 7.22 (d, J = 21.6Hz, 1H), 6.76 (d, J = 38.7Hz, 1H), 6.41 (s, 1H), 4.46-4.09 ( m, 3H), 4.04-3.82 (m, 2H), 3.67 (dd, J=21.6, 12.5Hz, 3H), 3.45 (dd, J=17.6, 7.0Hz, 1H), 2.13-1.97 (m, 2H), 1.30 (d, J=6.7Hz, 3H).

Example 12: Preparation of (S)-5-((1-(3-(2,2-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (12)

Step 1: Preparation of (S)-5-((1-(3-(2,2-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trifluoromethyl)ethoxy)methyl)pyridazin-3(2H)-one (12-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 0.087 mmol) was dissolved in N,N-dimethylformamide (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (50 mg, 0.13 mmol), N,N-diisopropylethylamine (41 μL, 0.235 mmol) and 2-(3,3-dimethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (31 mg, 0.104 mmol) were added and the reaction was carried out at 80°C overnight. Ethyl acetate was added to dilute, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product (S)-5-((1-(3-(2,2-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trifluoromethyl)ethoxy)methyl)pyridazin-3(2H)-one (12-a) (31 mg, yield 50.8%). ESI [M+H] ⁺ = 703.5.

Step 2: Preparation of (S)-5-((1-(3-(2,2-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (12)

(S)-5-((1-(3-(2,2-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trifluoromethyl)ethoxy)methyl)pyridazin-3(2H)-one (12-a) (31 mg, 0.044 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (3 ml) and reacted at room temperature for 3 hours. After concentration under reduced pressure, the reaction mixture was purified on a preparative plate (dichloromethane:methanol=15:1) to give the target product (S)-5-((1-(3-(2,2-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (12) (13.6 mg, yield 54.0%). ESI[M+H] ⁺ =573.4. ¹ H NMR (400MHz, CD ₃ OD) δ 8.61 (s, 2H), 7.39 (s, 1H), 7.08 (s, 1H), 6.73 (s, 1H), 6.24 (s, 1H), 4.35 (d, J = 4.9Hz, 1H), 4.19 (dd, J = 14.0, 3.2Hz, 1H), 3.99 (tt, J=14.0, 12.2Hz, 5H), 3.75 (d, J=5.3Hz, 2H), 1.54 (s, 6H), 1.35 (d, J=6.4Hz, 3H).

Example 13: Preparation of 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (13)

Step 1: Preparation of 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (13-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 0.087 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (50 mg, 0.13 mmol), N,N-diisopropylethylamine (40 μL, 0.226 mmol) and (S)-2-methyl-1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (27 mg, 0.096 mmol) were added and reacted at room temperature overnight. Water (5 ml) was added, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (13-a) (66 mg, yield 110.0%). ESI [M+H] ⁺ = 688.2.

Step 2: Preparation of 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (13)

5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (13-a) (66 mg, 0.096 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (4 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the reaction mixture was purified on a preparative plate (dichloromethane:methanol=15:1) to give the target product 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (13) (14 mg, yield 26.2%). ESI[M+H] ⁺ =558.2. ¹ H NMR (400MHz, CD ₃ OD) δ 8.37 (s, 1H), 7.75 (d, J = 9.0Hz, 1H), 7.42 (s, 1H), 7.15 (s, 1H), 6.88 (d, J = 9.1Hz, 1H), 6.79 (s, 1H), 6.32 (s, 1H), 4.66 (s, 2H), 4.46-3.86 (m, 7H), 3.29-3.20 (m, 1H), 1.36 (d, J=6.4Hz, 3H), 1.14 (d, J=5.7Hz, 3H).

Example 14: Preparation of 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (14)

Step 1: Preparation of 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (14-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (31 mg, 0.067 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (39 mg, 0.1 mmol), N,N-diisopropylethylamine (31 μL, 0.174 mmol) and (S)-2-(2-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrazine hydrochloride (21 mg, 0.074 mmol) were added and reacted at room temperature for 3 hours. Water (5 ml) was added, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (14-a) (32 mg, yield 69.6%). ESI [M+H] ⁺ = 689.1.

Step 2: Preparation of 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (14)

5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (14-a) (32 mg, 0.046 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (4 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 15: 1) to give the target product 5-((S)-1-(3-((S)-3-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (14) (16 mg, yield 61.5%). ESI[M+H] ⁺ =559.2. ¹ H NMR (400MHz, CD ₃ OD) δ 8.42 (s, 1H), 8.27 (s, 1H), 7.43 (s, 1H), 7.16 (s, 1H), 6.79 (s, 1H), 6.32 (s, 1H), 4.69 (d, J = 44.8Hz, 2H), 4.48 -4.11 (m, 5H), 3.97 (dd, J=13.7, 10.2Hz, 1H), 3.38 (d, J=11.8Hz, 2H), 1.36 (d, J=6.3Hz, 3H), 1.20 (d, J=5.4Hz, 3H).

Example 15: Preparation of (S)-5-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)pyrazine-2-carbonitrile (15)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (20 mg, 0.061 mmol) was dissolved in dichloromethane (4 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (35 mg, 0.091 mmol), N,N-diisopropylethylamine (27 μL, 0.153 mmol) and 5-(piperazin-1-yl)pyrazine-2-carbonitrile hydrochloride (14 mg, 0.061 mmol) were added and reacted at room temperature for 3 hours. After concentration under reduced pressure, the reaction mixture was purified on a preparative plate (dichloromethane:methanol=15:1) to give the target product (S)-5-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)pyrazine-2-carbonitrile (15) (3.9 mg, yield 12.8%). ESI[M+H] ⁺ =502.3, ESI[2M+H] ⁺ =1003.5. ¹ H NMR (400MHz, CD ₃ OD) δ 8.44 (s, 1H), 8.32 (s, 1H), 7.38 (s, 1H), 7.14 (s, 1H), 6.78 (s, 1H), 6.30 (s, 1H), 4.3 8 (s, 1H), 4.22 (d, J = 13.7Hz, 1H), 4.02-3.67 (m, 8H), 3.23 (d, J = 7.0Hz, 1H), 1.29 (s, 3H).

Example 16: Preparation of (S)-2-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)pyrimidine-5-carbonitrile (16)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (20 mg, 0.061 mmol) was dissolved in dichloromethane (4 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (35 mg, 0.091 mmol), N,N-diisopropylethylamine (27 μL, 0.153 mmol) and 2-(piperazin-1-yl)pyrimidine-5-carbonitrile hydrochloride (14 mg, 0.061 mmol) were added and reacted at room temperature for 3 hours. After concentration under reduced pressure, the reaction mixture was purified on a preparative plate (dichloromethane:methanol=15:1) to give the target product (S)-2-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)pyrimidine-5-carbonitrile (16) (12 mg, yield 39.5%). ESI[M+H] ⁺ =502.3, ESI[2M+H] ⁺ =1003.5. ¹ H NMR (400MHz, CD ₃ OD) δ 8.64 (s, 2H), 7.39 (s, 1H), 7.15 (s, 1H), 6.77 (s, 1H), 6.30 (s, 1H), 4.37 (s, 1H), 4.2 1(d, J=13.9Hz, 1H), 3.97(s, 4H), 3.76(s, 4H), 3.26-3.17(m, 1H), 1.30(s, 3H).

Example 17: Preparation of 4-(trifluoromethyl)-5-((2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclopentyl)amino)pyridazin-3(2H)-one (17)

Step 1: Preparation of 1-(2-hydroxycyclopentyl)-1H-pyrrole-3-carboxylic acid methyl ester (17-a)

Dissolve 3-pyrrolyl carboxylic acid methyl ester (1g, 8mmol) in N,N-dimethylformamide (20ml), add 1,2-epoxycyclopentane (4.2ml, 48mmol) and potassium carbonate (2.21g, 16mmol), heat in a sealed tube at 140℃ for 48 hours. Add water (50ml), extract with ethyl acetate, wash the organic phase with saturated brine, dry with anhydrous magnesium sulfate, filter, concentrate the filtrate under reduced pressure, and purify with a Flash column (ethyl acetate: petroleum ether = 10% to 30%) to obtain the target product 1-(2-hydroxycyclopentyl)-1H-pyrrole-3-carboxylic acid methyl ester (17-a) (1.48g, yield 88.6%). ESI[M+H] ⁺ = 210.1, ESI[2M+H] ⁺ = 419.3, ESI[2M+Na] ⁺ = 441.3.

Step 2: Preparation of 1-(2-(1,3-dihydroisoquinolin-2-yl)cyclopentyl)-1H-pyrrole-3-carboxylic acid methyl ester (17-b)

1-(2-Hydroxycyclopentyl)-1H-pyrrole-3-carboxylic acid methyl ester (17-a) (200 mg, 0.956 mmol) was dissolved in ultra-dry tetrahydrofuran (10 ml), phthalimide (169 mg, 1.15 mmol), triphenylphosphine (376 mg, 1.43 mmol) and diisopropyl azodicarboxylate (285 μL, 1.43 mmol) were added, and the mixture was heated at 70°C in a sealed tube to react overnight. After concentration under reduced pressure, the mixture was purified by preparative plate (ethyl acetate: petroleum ether = 1:2) to obtain the target product 1-(2-(1,3-dihydroisoquinolin-2-yl)cyclopentyl)-1H-pyrrole-3-carboxylic acid methyl ester (17-b) (515 mg). ESI[M+H] ⁺ = 339.2, ESI[2M+H] ⁺ = 677.4.

Step 3: Preparation of 1-(2-aminocyclopentyl)-1H-pyrrole-3-carboxylic acid methyl ester (17-c)

Dissolve 1-(2-(1,3-dihydroisoquinolin-2-yl)cyclopentyl)-1H-pyrrole-3-carboxylic acid methyl ester (17-b) (465 mg, 1.38 mmol) in methanol (15 ml), add hydrazine hydrate (1 ml, 13.75 mmol), and heat to 80°C in a sealed tube for 2 hours. After concentration under reduced pressure, purify on a preparative plate (dichloromethane: methanol = 10: 1) to obtain the target product 1-(2-aminocyclopentyl)-1H-pyrrole-3-carboxylic acid methyl ester (17-c) (94 mg, yield 53.1%). ESI [M+H] ⁺ = 209.1.

Step 4: Preparation of 1-(2-aminocyclopentyl)-1H-pyrrole-3-carboxylic acid (17-d)

Dissolve 1-(2-aminocyclopentyl)-1H-pyrrole-3-carboxylic acid methyl ester (17-c) (94 mg, 0.452 mmol) in methanol (5 ml), add water (1 ml) and sodium hydroxide (36 mg, 0.903 mmol), and react overnight at 80°C in a sealed tube. Use 1 equivalent of sodium hydroxide solution to adjust the pH to neutral, and concentrate under reduced pressure to obtain the target product 1-(2-aminocyclopentyl)-1H-pyrrole-3-carboxylic acid (17-d) (87 mg, yield 100.0%). ESI[M+H] ⁺ =195.2, ESI[2M+H] ⁺ =389.3.

Step 5: Preparation of 1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)cyclopentyl)-1H-pyrrole-3-carboxylic acid (17-e)

1-(2-Aminocyclopentyl)-1H-pyrrole-3-carboxylic acid (17-d) (87 mg, 0.448 mmol) was dissolved in ethanol (6 ml), and triethylamine (94 μL, 0.672 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (120 mg, 0.448 mmol) were added, and the mixture was reacted at 60° C. for 6 hours. After concentration under reduced pressure, the mixture was purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product 1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)cyclopentyl)-1H-pyrrole-3-carboxylic acid (17-e) (96 mg, yield 44.0%). ESI[M+H] ⁺ =487.3, ESI[2M+H] ⁺ =973.5.

Step 6: Preparation of 4-(trifluoromethyl)-5-((2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclopentyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (17-f)

1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)cyclopentyl)-1H-pyrrole-3-carboxylic acid (17-e) (47 mg, 0.097 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (55 mg, 0.145 mmol), N,N-diisopropylethylamine (44 μL, 0.252 mmol) and 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (26 mg, 0.097 mmol) were added and reacted at room temperature for 4 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product 4-(trifluoromethyl)-5-((2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclopentyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (17-f) (69 mg, yield 101.5%). ESI [M+H] ⁺ = 701.5.

Step 7: Preparation of 4-(trifluoromethyl)-5-((2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclopentyl)amino)pyridazin-3(2H)-one (17)

4-(Trifluoromethyl)-5-((2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclopentyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (17-f) (69 mg, 0.099 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (5 ml) and reacted at room temperature for 4 hours. After concentration under reduced pressure, the mixture was purified by preparative plate (dichloromethane:methanol=15:1) to obtain the target product 4-(trifluoromethyl)-5-((2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclopentyl)amino)pyridazin-3(2H)-one (17) (41.7 mg, yield 74.2%). ESI[M+H] ⁺ =571.3. ¹ H NMR (400MHz, CD ₃ OD) δ 8.60 (s, 2H), 7.81 (s, 1H), 7.17 (t, J=1.7Hz, 1H), 6.81 (t, J=2.5Hz, 1H), 6.31 (dd, J=2.6, 1.8Hz, 1H), 5.65 (dd, J=8.0, 4.6Hz, 1H), 4.77 (dd, J=15.4, 7. 8Hz, 1H), 4.51 (dd, J=12.9, 6.6Hz, 1H), 4.00-3.90 (m, 4H), 3.79-3.71 (m, 4H), 2.42-2.30 (m, 2H), 2.27-1.97 (m, 3H), 1.94-1.65 (m, 3H).

Example 18: Preparation of 4-(trifluoromethyl)-5-((4-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)tetrahydrofuran-3-yl)amino)pyridazin-3(2H)-one (18)

Step 1: Preparation of 1-(4-hydroxytetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid methyl ester (18-a)

Dissolve 3-pyrrolyl carboxylic acid methyl ester (600 mg, 4.8 mmol) in N, N-dimethylformamide (15 ml), add 3, 4-epoxytetrahydrofuran (2 ml, 28.8 mmol) and potassium carbonate (1.32 g, 9.6 mmol), heat in a sealed tube at 110°C for 24 hours. Add water (20 ml), extract with ethyl acetate, wash the organic phase with saturated brine, dry with anhydrous magnesium sulfate, filter, concentrate the filtrate under reduced pressure, and purify with a Flash column (ethyl acetate: petroleum ether-10% to 30%) to obtain the target product 1-(4-hydroxytetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid methyl ester (18-a) (715 mg, yield 65.2%). ESI[M+H] ⁺ =211.9, ESI[2M+H] ⁺ =423.2.

Step 2: Preparation of 1-(4-(1,3-dihydroisoquinolin-2-yl)tetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid methyl ester (18-b)

Dissolve 1-(4-hydroxytetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid methyl ester (18-a) (300 mg, 1.42 mmol) in ultra-dry tetrahydrofuran (15 ml), add phthalimide (251 mg, 1.71 mmol), triphenylphosphine (559 mg, 2.13 mmol) and diisopropyl azodicarboxylate (422 μL, 2.13 mmol), and heat to 70°C in a sealed tube for 6 hours. After concentration under reduced pressure, the target product 1-(4-(1,3-dihydroisoquinolin-2-yl)tetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid methyl ester (18-b) (420 mg, yield 65.2%) was obtained. ESI[M+H] ⁺ =341.1, ESI[2M+H] ⁺ =681.2.

Step 3: Preparation of 1-(4-aminotetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid methyl ester (18-c)

Dissolve 1-(4-(1,3-dihydroisoquinolin-2-yl)tetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid methyl ester (18-b) (420 mg, 1.24 mmol) in methanol (20 ml), add hydrazine hydrate (770 μL, 12.35 mmol), and heat to 80°C in a sealed tube for 2 hours. After concentration under reduced pressure, the residue was dissolved in dichloromethane, and the insoluble solid was filtered out. After the filtrate was concentrated under reduced pressure, it was purified by Flash column (methanol: dichloromethane = 0% to 10%) to obtain the target product 1-(4-aminotetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid methyl ester (18-c) (240 mg, yield 92.7%). ESI[M+H] ⁺ = 211.0, ESI[2M+H] ⁺ = 421.3.

Step 4: Preparation of 1-(4-aminotetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid (18-d)

Dissolve 1-(4-aminotetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid methyl ester (18-c) (240 mg, 1.14 mmol) in methanol (15 ml), add water (3 ml) and sodium hydroxide (92 mg, 2.28 mmol), and react overnight at 80°C in a sealed tube. Use 1 equivalent of sodium hydroxide solution to adjust the pH to neutral, and concentrate under reduced pressure to obtain the target product 1-(4-aminotetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid (18-d) (344 mg, yield 153.6%). ESI [M+H] ⁺ = 197.1.

Step 5: Preparation of 1-(4-((6-oxy-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)tetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid (18-e)

1-(4-aminotetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid (18-d) (172 mg, 0.57 mmol, 65%) was dissolved in ethanol (5 ml), triethylamine (119 μL, 0.86 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (187 mg, 0.57 mmol) were added, and the mixture was reacted at 60° C. for 4 hours. After concentration under reduced pressure, the mixture was purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product 1-(4-((6-oxyl-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)tetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid (18-e) (40 mg, yield 14.4%). ESI[M+H] ⁺ =489.3, ESI[2M+H] ⁺ =977.3.

Step 6: Preparation of 4-(trifluoromethyl)-5-((4-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)tetrahydrofuran-3-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (18-f)

1-(4-((6-oxyl-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)tetrahydrofuran-3-yl)-1H-pyrrole-3-carboxylic acid (18-e) (40 mg, 0.082 mmol) was dissolved in dichloromethane (5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (47 mg, 0.123 mmol), N,N-diisopropylethylamine (37 μL, 0.213 mmol) and 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (22 mg, 0.082 mmol) were added and reacted at room temperature for 2 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product 4-(trifluoromethyl)-5-((4-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)tetrahydrofuran-3-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (18-f) (50 mg, yield 87.0%). ESI [M+H] ⁺ = 703.3.

Step 7: Preparation of 4-(trifluoromethyl)-5-((4-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)tetrahydrofuran-3-yl)amino)pyridazin-3(2H)-one (18)

4-(Trifluoromethyl)-5-((4-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)tetrahydrofuran-3-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (18-f) (50 mg, 0.071 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (4 ml) and reacted at room temperature for 4 hours. After concentration under reduced pressure, the mixture was purified on a preparative plate (dichloromethane:methanol=15:1) to obtain the target product 4-(trifluoromethyl)-5-((4-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)tetrahydrofuran-3-yl)amino)pyridazin-3(2H)-one (18) (27 mg, yield 66.3%). ESI[M+H] ⁺ =573.2, ESI[2M+H] ⁺ =1145.3. ¹ H NMR (600MHz, DMSO-d ₆ ) δ 12.55 (s, 1H), 8.74 (s, 2H), 7.91 (s, 1H), 7.18 (s, 1H), 6.81 (s, 1H), 6.21 (s, 1H), 5 .86(s, 1H), 5.09(s, 1H), 4.82(s, 1H), 4.20-4.00(m, 4H), 3.84(s, 4H), 3.61(s, 4H).

Example 19: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(trifluoromethyl)phenyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (19)

Step 1: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (19-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 1 eq.), N,N-diisopropylethylamine (34 mg, 3 eq.) and 1-(4-(trifluoromethyl)phenyl)piperazine (20 mg, 1.2 eq.) were added and reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and then purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)phenyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (19-a) (55 mg, yield 94.11%). ESI [M+H] ⁺ = 673.3.

Step 2: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)phenyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (19)

(S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)phenyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (19-a) (55 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 3 hours. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to give the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)phenyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (19) (15.3 mg, yield 34.50%). ESI[M+H] ⁺ =543.3. ¹ H NMR (600MHz, CD ₃ OD) δ7.49 (d, J=8.4Hz, 2H), 7.36 (s, 1H), 7.11-7.05 (m, 3H), 6.78 (s, 1H), 6.29 (s, 1H), 4.37 (s, 1H), 4.21 (d, J=1 4.1Hz, 1H), 3.98-3.90 (m, 1H), 3.78 (dd, J=15.3, 9.8Hz, 5H), 3.31 (s, 2H), 1.36 (d, J=6.3Hz, 3H).

Example 20: Preparation of 4-(trifluoromethyl)-5-((1R,2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)pyridazin-3(2H)-one (20)

Step 1: Preparation of tert-butyl ((1R, 2S)-2-hydroxycyclohexyl) carbamate (20-a)

Under ice bath, triethylamine (1.04mL, 8.90mmol) was added to a dichloromethane solution (30ml) of (1S, 2R)-2-aminocyclohexane-1-ol hydrochloride (540mg, 3.56mmol), and then di-tert-butyl dicarbonate (1.3ml, 4.27mmol) was added dropwise. After stirring for 1 hour under ice bath, the mixture was heated to room temperature and continued to stir for 3 hours. The reaction solution was quenched with water (50mL), extracted with dichloromethane (20mLx3), and the organic phase was washed with salt water (20mL), dried over anhydrous magnesium sulfate, filtered and concentrated. The crude product of tert-butyl ((1R, 2S)-2-hydroxycyclohexyl) carbamate (20-a) (1.45g) was obtained.

Step 2: Preparation of (1S, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl 4-toluenesulfonate (20-b)

Under ice bath, triethylamine (1.5mL, 10.78mmol) and 4-dimethylaminopyridine (1.32g, 10.78mmol) were added to a dichloromethane solution (30mL) of tert-butyl ((1R, 2S)-2-hydroxycyclohexyl) carbamate (20-a) (1.45g crude product, 5.39mmol), and stirred under ice bath for 10 minutes. Then, p-toluenesulfonyl chloride (1.54g, 8.09mmol) was added, and the reaction solution was stirred at room temperature for 4 hours. The reaction solution was quenched with water (50mL), extracted with dichloromethane (20mLx3), and the organic phase was washed with brine (20mL), dried over anhydrous magnesium sulfate, filtered and concentrated. The crude product of (1S, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl 4-toluenesulfonate (1.63g) (20-b) was obtained.

Step 3: Preparation of methyl 1-((1R, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylate (20-c)

At room temperature, cesium carbonate (2.52 g, 7.71 mmol) was added to a solution of 1H-pyrrole-3-carboxylic acid methyl ester (20-b) (322 mg, 2.57 mmol) in N, N-dimethylformamide (25 ml), and stirred at room temperature for 30 minutes. (1S, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl 4-toluenesulfonate (1.63 g crude product, 3.09 mmol) was added, and the reaction solution was stirred at 80 degrees overnight. The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (30 mL x 3), and the organic phase was washed with brine (30 mL x 3), dried over anhydrous magnesium sulfate, filtered and concentrated. The crude product was purified by silica gel chromatography (petroleum ether: ethyl acetate = 2: 1) to give 1-((1R, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid methyl ester (20-c) (90 mg, yield: 10.86%). ESI [M+H] ⁺ = 323.2.

Step 4: Preparation of 1-((1R, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid (20-d)

At room temperature, sodium hydroxide (22.4 mg, 0.558 mmol) and water (3 mL) were added to a solution of 1-((1R, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid methyl ester (20-c) (90 mg, 0.2.79 mmol) in methanol (20 mL), and the reaction solution was reacted overnight at 80 degrees in a vacuum tube. The reaction solution was subjected to reduced pressure distillation to remove the solvent, and water (20 mL) was added. The pH was adjusted to 2 with 4M hydrochloric acid solution, and 1-((1R, 2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid (20-d) (65 mg, yield: 75.51%) was obtained by filtration. ESI [M+H] ⁺ = 309.2.

Step 5: Preparation of tert-butyl ((1R, 2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)carbamate (20-e)

To a solution of 1-((1R,2R)-2-((tert-butyloxycarbonyl)amino)cyclohexyl)-1H-pyrrole-3-carboxylic acid (20-d) (65 mg, 0.211 mmol) in dichloromethane (10 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (120 mg, 0.317 mmol), N,N-diisopropylethylamine (110 μL, 0.633 mmol), and 2-(piperazine- 1-yl)-5-(trifluoromethyl)pyrimidine (49 mg, 0.211 mmol), stirred at room temperature for 4 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (dichloromethane: methanol = 10: 1) to obtain tert-butyl ((1R, 2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)carbamate (20-e) (32 mg, yield: 29.05%). ESI [M+H] ⁺ = 523.2.

Step 6: Preparation of (1-((1R, 2R)-2-aminocyclohexyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (20-f)

At room temperature, a 4M hydrochloric acid dioxane solution (2 mL) was added to a solution of tert-butyl ((1R, 2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl (20-e) (34 mg, 0.065 mmol) in ethyl acetate (2 mL). The reaction solution was stirred at room temperature for 2 hours and then concentrated under reduced pressure to give (1-((1R, 2R)-2-aminocyclohexyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (20-f) (32 mg, crude product). ESI [M+H] ⁺ = 423.2.

Step 7: Preparation of 4-(trifluoromethyl)-5-((1R, 2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (20-g)

To a solution (5 mL) of 4-(trifluoromethyl)-5-((1R,2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (20-f) (32 mg, crude, 0.076 mmol) in ethanol was added triethylamine (34 μL, 0.265 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one at room temperature. The reaction solution was stirred at 60 degrees for 4 hours and then concentrated under reduced pressure. The crude product was purified by TLC (dichloromethane: methanol = 15: 1) to give 4-(trifluoromethyl)-5-((1R, 2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazine-3(2H)-one (20-g) (7 mg, yield 12.93%). ESI [M+H] ⁺ = 715.2.

Step 8: Preparation of 4-(trifluoromethyl)-5-((1R, 2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (20)

At room temperature, 4-(trifluoromethyl)-5-((1R,2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (20-g) (7 mg, 0.0098 mmol) was dissolved in 4M hydrochloric acid ethyl acetate (2 mL) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative high performance liquid chromatography to give (4-(trifluoromethyl)-5-((1R, 2R)-2-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)cyclohexyl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (20) (3.2 mg, yield: 55.9%), ESI [M+H ⁺ ]=585.3. ¹ H NMR (400MHz, CD ₃ OD) δ 8.59 (s, 2H), 7.56 (s, 1H), 7.20 (s, 1H), 6.82 (s, 1H), 6.24 (s, 1H), 4.05 (s, 1H), 3.93 (d, J=5.3Hz, 4H), 3.80-3.63 (m , 4H), 2.23-1.96 (m, 4H), 1.70-1.44 (m, 4H), 1.28 (s, 1H).

Example 21: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (21)

Step 1: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (21-a)

To a solution of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 0.087 mmol) in dichloromethane (5 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (49.5 mg, 0.130 mmol), N,N-diisopropylethylamine (45 μL, 0.261 mmol), and (S)-2-(3-methylpiperazin-1-yl)- 5-(Trifluoromethyl)pyrimidine (21.4 mg, 0.087 mmol) was stirred at room temperature for 4 hours, and then the reaction solution was concentrated under reduced pressure. The crude product was purified by TLC preparation plate (dichloromethane: methanol = 10: 1) to give 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (21-a) (38 mg, yield 63.54%). ESI [M+H] ⁺ = 689.2.

Step 2: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (21)

5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (21-a) (38 mg, 0.055 mmol) was dissolved in 4 M hydrochloric acid and ethyl acetate (4 m L), stirred at room temperature for 2 hours, the reaction solution was concentrated under reduced pressure, and the crude product was slurried with ethyl acetate (0.5 mL) to obtain 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (21) (19 mg, yield 61.6%). ESI[M+H ⁺ ]＝559.3. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.59 (s, 2H), 7.43 (s, 1H), 7.15 (s, 1H), 6.78-6.75 (m, 1H), 6.30 (s, 1H), 5.48 (s, 1H), 5.19 (s, 1H), 4.37 (s, 1H), 4.32-4.27 (m, 1H), 4.19 (dd, J=1 4.1, 3.7Hz, 2H), 3.97 (dd, J=14.1, 9.8Hz, 2H), 3.49-3.44 (m, 1H), 2.05-1.97 (m, 1H), 1.59-1.52 (m, 1H), 1.34 (d, J=6.5Hz, 3H), 1.17 (d, J=5.7Hz, 3H).

Example 22: Preparation of 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (22)

Step 1: Preparation of 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (22-a)

To a solution of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (31 mg, 0.067 mmol) in dichloromethane (5 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (38.2 mg, 0.101 mmol), N,N-diisopropylethylamine (35 μL, 0.201 mmol), and (R)-2-(3-methylpiperazin-1-yl)- 5-(Trifluoromethyl)pyrimidine (16.5 mg, 0.067 mmol) was stirred at room temperature for 4 hours, and then the reaction solution was concentrated under reduced pressure. The crude product was purified by TLC preparation plate (dichloromethane: methanol = 10: 1) to give 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (22-a) (20 mg, yield 43.33%). ESI [M+H] ⁺ = 689.2.

Step 2: Preparation of 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (22)

At room temperature, 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (22-a) (20 mg, 0.029 mmol) was dissolved in 4 M hydrochloric acid and ethyl acetate. (4 mL), stirred at room temperature for 2 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative high performance liquid chromatography to give 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (22) (3 mg, yield 18.5%). ESI[M+H ⁺ ]＝559.3. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.57 (s, 2H), 7.37 (d, J = 6.1Hz, 1H), 7.11 (s, 1H), 6.76 (t, J = 2.4Hz, 1H), 6.27 (s, 1H), 4.76 (d, J = 13.4Hz, 1H), 4.68 (d, J = 13.3Hz, 2H), 4.36 (s, 2H), 4 .25(d,J=1 1.2Hz, 1H), 4.19 (dd, J=14.1, 3.6Hz, 1H), 3.94 (dd, J=14.1, 9.9Hz, 1H), 3.10 (d, J=10.0Hz, 2H), 1.55 (d, J=7.0Hz, 1H), 1.34 (d, J=6.5Hz, 3H), 1.21 (t, J=6. 1Hz, 3H).

Example 23: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (23)

Step 1: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidin-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (23-a)

To a solution of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (41.8 mg, 0.091 mmol) in dichloromethane (5 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (52 mg, 0.136 mmol), N,N-diisopropylethylamine (47.4 μL, 0.272 mmol), and 2-(piperidin-3-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (41.8 mg, 0.091 mmol). After stirring at room temperature for 4 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by TLC preparation plate (ethyl acetate) to obtain (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidin-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (23-a) (5 mg, yield 8.17%). ESI[M+H] ⁺ =674.2.

Step 2: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (23)

At room temperature, (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (23-a) (5 mg, 0.074 mmol) was dissolved in 4M hydrochloric acid ethyl acetate (2 mL) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and the crude product was purified by preparative high performance liquid chromatography to give (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (23) (1.8 mg, yield 44.63%). ESI[M+H] ⁺ ＝544.3.

Example 24: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)cyclohexyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (24)

Step 1: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)cyclohexyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (24-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 1 eq.), N,N-diisopropylethylamine (34 mg, 3 eq.) and 1-(4-(trifluoromethyl)cyclohexyl)piperazine (20 mg, 1 eq.) were added and reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and then purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)cyclohexyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (24-a) (20 mg, yield 33.92%). ESI [M+H] ⁺ = 579.3.

Step 2: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)cyclohexyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (24)

5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (24-a) (20 mg, 1 eq.) was dissolved in 4 M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 3 hours. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to give the target products (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)cyclohexyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (6.3 mg, yield 38.98%) and (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)cyclohexyl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (24) (3.6 mg, yield 22.27%). ESI[M+H] ⁺ =549.3. ¹ H NMR (600MHz, CD ₃ OD) δ7.40 (s, 1H), 7.14 (s, 1H), 6.79 (s, 1H), 6.28 (s, 1H), 4.44 (d, J = 71.5Hz, 4H), 4.22 (d, J = 14.1Hz, 1H), 3.97-3. 90 (m, 1H), 3.67-3.54 (m, 2H), 3.09 (d, J = 17.9Hz, 2H), 2.48 (s, 1H), 2.26 (d, J = 11.1Hz, 1H), 2.19-1.98 (m, 5H), 1.93-1.73 (m, 4H), 1.36 (d, J = 6.3Hz, 3H) . ¹ H NMR (600MHz, CD ₃ OD) δ7.40 (s, 1H), 7.13 (s, 1H), 6.80 (s, 1H), 6.28 (s, 1H), 4.57-4.33 (m, 4H), 4.22 (d, J=14.1Hz, 2H), 3.99-3.87 (m, 2H), 2.26 (d, J= 11.8Hz, 4H), 2.15 (d, J=13.0Hz, 3H), 2.03 (d, J=12.4Hz, 1H), 1.89-1.75 (m, 2H), 1.61 (d, J=10.5Hz, 3H), 1.36 (d, J=6.4Hz, 3H).

Example 25: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (25)

Step 1: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (25-a)

To a solution of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (30 mg, 0.065 mmol) in dichloromethane (5 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (37.1 mg, 0.098 mmol), N,N-diisopropylethylamine (34 μL, 0.195 mmol), and 2-(piperazine-1 -yl)-4-(trifluoromethyl)pyrimidine (15.1 mg, 0.065 mmol), stirred at room temperature for 4 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by TLC preparation plate (ethyl acetate) to obtain (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (25-a) (27 mg, yield 61.54%). ESI[M+H] ⁺ =675.2.

Step 2: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (25)

At room temperature, (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperidine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (25-a) (27 mg, 0.04 mmol) was dissolved in 4M hydrochloric acid ethyl acetate (2 mL) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and the crude product was purified by preparative high performance liquid chromatography to give (S)-4-(trifluoromethyl)-5-((1-(3-(4-(4-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (25) (6.2 mg, yield 28.46%). ESI [M + H ⁺ ] = 545.4. ¹ H NMR (400MHz, CD ₃ OD) δ 8.60 (d, J = 4.8Hz, 1H), 7.39 (s, 1H), 7.15 (s, 1H), 6.92 (d, J = 4.8Hz, 1H), 6.77-6.73 (m, 1H), 6.30 (d, J = 9.6Hz, 1H ), 4.36 (s, 1H), 4.19 (dd, J=14.1, 3.7Hz, 1H), 3.95 (dd, J=14.9, 10.6Hz, 2H), 3.92-3.84 (m, 4H), 3.81-3.73 (m, 4H), 1.33 (t, J=6.0Hz, 3H).

Example 26: Preparation of (S)-2-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)pyrimidine-4-carbonitrile (26)

To a solution of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (20 mg, 0.061 mmol) in dichloromethane (5 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (34.5 mg, 0.091 mmol), N,N-diisopropylethylamine (31.6 μL, 0.182 mmol), and 2-(piperazine)-1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (20 mg, 0.061 mmol) in dichloromethane (5 mL). -1-yl)pyrimidine-4-carbonitrile (11.5 mg, 0.061 mmol), after stirring at room temperature for 4 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by TLC preparation plate (dichloromethane: methanol = 10: 1) to give (S)-2-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)pyrimidine-4-carbonitrile (26) (20 mg, yield 62.64%). ESI [M + H ⁺ ] = 502.3. ¹ H NMR (400MHz, CD ₃ OD) δ 8.56 (d, J = 4.7Hz, 1H), 7.38 (s, 1H), 7.14 (s, 1H), 6.96 (d, J = 4.7Hz, 1H), 6.75 (d, J = 2.2Hz, 1H), 6.29 (s, 1H), 4.6 9-4.57 (m, 1H), 4.42-4.32 (m, 1H), 4.25-4.15 (m, 1H), 3.89 (s, 2H), 3.87 (s, 4H), 3.73 (dd, J=19.9, 6.0Hz, 4H), 1.31 (d, J=23.0Hz, 3H).

Example 27: Preparation of 5-((S)-1-(3-((S)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (27)

Step 1: Preparation of 5-((S)-1-(3-((S)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (27-a)

To a solution of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (20 mg, 0.044 mmol) in N,N-dimethylformamide (5 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (25 mg, 0.066 mmol), N,N-diisopropylethylamine (23 μL, 0.350 mmol), and (S)-2-(3-isopropylpiperazin-1-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (20 mg, 0.044 mmol). )-5-(trifluoromethyl)pyrimidine (12 mg, 0.044 mmol), stirred at 80 degrees Celsius overnight, the reaction solution was concentrated under reduced pressure, and the crude product was purified by TLC preparation plate (dichloromethane: methanol = 10: 1) to obtain 5-((S)-1-(3-((S)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (27-a) (12 mg, yield 38.27%). ESI [M+H] ⁺ = 717.2.

Step 2: Preparation of 5-((S)-1-(3-((S)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (27)

At room temperature, 5-((S)-1-(3-((S)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (27-a) (12 mg, 0.017 mmol) was dissolved in 4 M hydrochloric acid and ethyl acetate. (2 mL), stirred at room temperature for 2 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative high performance liquid chromatography to give 5-((S)-1-(3-((S)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (27) (5 mg, yield 50.9%). ESI[M+H] ⁺ =587.1 ¹ H NMR (400 MHz, CD ₃ OD)δ8.57(s,2H),7.39-7.29(m,1H),7.15-7.10(m,1H),6.79-6.73(m,1H),6.30-6.24(m,1H),4.44-4.33(m,2H),4.25-4.20(m,1H),4.20-4.16(m ,1H ), 4.12-4.07(m, 1H), 4.06-4.01(m, 1H), 4.00-3.89(m, 2H), 3.63-3.56(m, 1H), 3.52-3.45(m, 1H), 3.17-3.06(m, 2H), 1.28(s, 6H), 1.16-1.02(m, 3 H).

Example 28: Preparation of 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (28)

Step 1: Preparation of 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (28-a)

To a solution of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (30 mg, 0.065 mmol) in dichloromethane (5 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (37.1 mg, 0.098 mmol), N,N-diisopropylethylamine (34 μL, 0.195 mmol), and (R)-3-methyl-1-(5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (30 mg, 0.065 mmol). After stirring at room temperature for 4 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by TLC preparation plate (ethyl acetate) to obtain 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (28-a) (12 mg, yield 26.74%). ESI[M+H] ⁺ =688.2.

Step 2: Preparation of 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (28)

At room temperature, 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (28-a) (12 mg, 0.017 mmol) was dissolved in 4 M hydrochloric acid ethyl acetate ( 2mL), stirred at room temperature for 2 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative high performance liquid chromatography to give 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (28) (2.8 mg, yield 28.8%). ESI[M+H] ⁺ =558.3. ¹ H NMR (600 MHz, CD ₃ OD) δ 8.34 (s, 1H), 7.74 (d, J = 8.6Hz, 1H), 7.34 (s, 1H), 7.09 (s, 1H), 6.91 (d, J = 9.1Hz, 1H), 6.78 (s, 1H), 6.28 (s, 1H), 4.64-4.56 (m, 1H), 4.41-4.35 (m ,1H),4. 35-4.29 (m, 1H), 4.26-4.18 (m, 2H), 3.97-3.91 (m, 1H), 3.47-3.38 (m, 2H), 3.22-3.13 (m, 1H), 3.09-3.02 (m, 1H), 1.36 (d, J=6.3Hz, 3H), 1.25 (d, J=6 .7Hz, 3H).

Example 29: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (29)

Step 1: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (29-a)

To a solution of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (30 mg, 0.065 mmol) in dichloromethane (5 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (37.1 mg, 0.098 mmol), N,N-diisopropylethylamine (34 μL, 0.195 mmol), and (S)-3-methyl-1-(5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (30 mg, 0.065 mmol). After stirring at room temperature for 4 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by TLC preparation plate (ethyl acetate) to obtain 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (29-a) (21 mg, yield 26.74%). ESI[M+H] ⁺ =688.2.

Step 2: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (29)

5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (29-a) (21 mg, 0.031 mmol) was dissolved in 4 M hydrochloric acid ethyl ester (2 M) at room temperature. mL), stirred at room temperature for 2 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative high performance liquid chromatography to give 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (29) (3.4 mg, yield 19.97%). ESI[M+H ⁺ ]＝558.3. ¹ H NMR (600 MHz, CD ₃ OD) δ8.34 (s, 1H), 7.74 (d, J=8.8Hz, 1H), 7.39 (s, 1H), 7.09 (s, 1H), 6.91 (d, J=9.1Hz, 1H), 6.78 (s, 1H), 6.27 (s, 1H), 4.71-4.58 (m, 2H), 4.37 (s, 2H), 4.34-4.27 (m, 1H), 4.25-4.17 (m, 2H), 3.99-3.91 (m, 1H), 3.42 (s, 2H), 3.05 (s, 1H), 2.85-2.79 (m, 1H), 1.36 (d, J=6.4Hz, 3H), 1.25 (d, J=6.7Hz, 3H) .

Example 30: Preparation of 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (30)

Step 1: Preparation of 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (30-a)

To a solution of (S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (30 mg, 0.065 mmol) in dichloromethane (5 mL) at room temperature were added 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (37.1 mg, 0.098 mmol), N,N-diisopropylethylamine (34 μL, 0.195 mmol), and (R)-2-(3-methylpiperazine-1 -yl)-5-(trifluoromethyl)pyrazine (16.1 mg, 0.065 mmol), stirred at room temperature for 4 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by TLC preparation plate (ethyl acetate) to obtain 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrazine-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (30-a) (20 mg, yield 44.41%). ESI[M+H] ⁺ =689.2.

Step 2: Preparation of 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (30)

5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (30-a) (20 mg, 0.030 mmol) was dissolved in 4 M hydrochloric acid ethyl ester (2 M) at room temperature. mL), stirred at room temperature for 2 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative high performance liquid chromatography to give 5-((S)-1-(3-((R)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (30) (5.1 mg, yield 31.34%). ESI[M+H] ⁺ =559.2. ¹ H NMR (600 MHz, CD ₃ OD) δ 8.39 (s, 1H), 8.30 (s, 1H), 7.35 (s, 1H), 7.11 (s, 1H), 6.77 (s, 1H), 6.28 (s, 1H), 4.65 (s, 2H), 4.37 (s, 1H), 4.33 (d, J=13.8Hz, 2H), 4.25 (s, 1H), 4.21 (d, J=14.2Hz, 2H), 3.94 (dd, J=13.9, 10.3Hz, 1H), 3.14 (t, J=10.8Hz, 2H), 1.36 (d, J=6.4Hz, 3H), 1.26 (d, J=6.7Hz, 3H).

Example 31: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (31)

Step 1: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (31-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 1 eq.), N,N-diisopropylethylamine (34 mg, 3 eq.) and (S)-2-(3-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrazine (26 mg, 1.2 eq.) were added and reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and then purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (31-a) (30 mg, yield 50.15%). ESI [M+H] ⁺ = 689.2.

Step 2: Preparation of 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (31)

5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (31-a) (30 mg, 1 eq.) was dissolved in 4 M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 3 hours. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to give the target product 5-((S)-1-(3-((S)-2-methyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (31) (8.5 mg, yield 34.94%). ESI[M+H] ⁺ =559.2. ¹ H NMR (600MHz, CD ₃ OD) δ8.39 (s, 1H), 8.30 (s, 1H), 7.39 (s, 1H), 7.10 (s, 1H), 6.78 (s, 1H), 6.27 (s, 1H), 4.44-4.29 (m, 4H), 4.21 (d, J=14.1Hz, 2H), 3.97-3.92 (m, 1H), 3.42 (s, 2H), 3.14 (t, J=11.5Hz, 1H), 1.35 (d, J=6.4Hz, 3H), 1.26 (d, J=6.6Hz, 3H).

Example 32: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (32)

Step 1: Preparation of (1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (32-a)

1H-indole-3-carboxylic acid (100 mg, 1 eq.) was dissolved in dichloromethane (8 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (236 mg, 1 eq.), N,N-diisopropylethylamine (241 mg, 3 eq.) and 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (150 mg, 1.2 eq.) were added, and the mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and purified by Flash column (petroleum ether: ethyl acetate = 1:1) to obtain the target product (1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidine-2-yl)piperazine-1-yl)methanone (32-a) (150 mg, yield 64.40%). ESI [M+H] ⁺ = 376.2.

Step 2: Preparation of tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)carbamate (32-b)

(1H-Indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (32-a) (150 mg, 1 eq.) was dissolved in N,N-dimethylformamide (8 ml), and cesium carbonate (390 mg, 3 eq.) and tert-butyl-(1-iodopropan-2-yl)carbamate (170 mg, 1.5 eq.) were added, and the reaction was carried out at 80°C overnight. Water (80 ml) was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by Flash column (petroleum ether: ethyl acetate = 1: 1) to obtain the target product tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)carbamate (32-b) (90 mg, yield 42.29%). ESI [M+H] ⁺ = 533.4.

Step 3: Preparation of (S)-(1-(2-aminopropyl)-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (32-c)

Tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)carbamate (32-b) (90 mg, 1 eq.) was dissolved in dichloromethane (1 ml), trifluoroacetic acid (1 ml) was added, and the mixture was reacted at room temperature for 1 hour. After concentration under reduced pressure, the target product (S)-(1-(2-aminopropyl)-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (32-c) (120 mg, purity 50%, yield 82.10%) was obtained. ESI[M+H] ⁺ =433.3.

Step 4: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (32-d)

(S)-(1-(2-Aminopropyl)-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (32-c) (120 mg, purity 50%, 1 eq.) was dissolved in ethanol (2 ml), and triethylamine (42 mg, 3 eq.) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (55 mg, 1.2 eq.) were added, and the reaction was carried out at 60°C for 16 hours. After concentration under reduced pressure, the mixture was purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to give the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (32-d) (41 mg, yield 40.77%). ESI [M+H] ⁺ = 725.5

Step 5: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (32)

(S)-4-(Trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (32-d) (41 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (3 ml) and reacted at room temperature for 6 hours. After concentration under reduced pressure, the mixture was purified on a preparative plate (dichloromethane: methanol = 10: 1) to give the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (32) (14.9 mg, yield 44.30%). ESI [M+H] ⁺ = 595.4. ¹ H NMR (600 MHz, CD ₃ OD) δ8.61 (s, 2H), 7.66 (s, 1H), 7.62 (dt, J=15.7, 7.7Hz, 2H), 7.29 (t, J=7.7Hz, 1H), 7.19 (t, J=7.6Hz, 1H), 7.05 (s, 1H), 4.31 (dd, J=14.5, 10.0Hz, 1H), 4 .00 (dd, J=11.5, 6.2Hz, 4H), 3.95 (dd, J=8.2, 6.3Hz, 1H), 3.81-3.74 (m, 4H), 2.04 (dd, J=12.3, 6.4Hz, 1H), 1.46 (t, J=6.2Hz, 3H).

Example 33: Preparation of (S)-5-((1-(6-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (33)

Step 1: Preparation of (6-fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (33-a)

6-Fluoro-1H-indole-3-carboxylic acid (100 mg, 1 eq.) was dissolved in dichloromethane (8 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (212 mg, 1 eq.), N,N-diisopropylethylamine (2216 mg, 3 eq.) and 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (156 mg, 1.2 eq.) were added, and the mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and purified by Flash column (petroleum ether: ethyl acetate = 1:1) to obtain the target product (6-fluoro-1H-indole-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (33-a) (77 mg, yield 35.07%). ESI [M+H] ⁺ = 394.20.

Step 2: Preparation of tert-butyl-(1-(6-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)carbamate (33-b)

(6-Fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (33-a) (77 mg, 1 eq.) was dissolved in N,N-dimethylformamide (5 ml), and cesium carbonate (191 mg, 3 eq.) and tert-butyl-(1-iodopropan-2-yl)carbamate (73 mg, 1.3 eq.) were added, and the reaction was carried out at 80°C overnight. Water (30 ml) was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by Flash column (petroleum ether: ethyl acetate = 4: 1) to obtain the target product tert-butyl-(1-(6-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)carbamate (33-b) (100 mg, yield 92.79%). ESI [M+H] ⁺ = 551.4.

Step 3: Preparation of (S)-(1-(2-aminopropyl)-6-fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (33-c)

Tert-butyl-(1-(6-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)carbamate (33-b) (100 mg, 1 eq.) was dissolved in dichloromethane (1 ml), trifluoroacetic acid (1 ml) was added, and the mixture was reacted at room temperature for 1 hour. After concentration under reduced pressure, the target product (S)-(1-(2-aminopropyl)-6-fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (33-c) (120 mg, purity 60%, yield 88.00%) was obtained. ESI[M+H] ⁺ =451.3.

Step 4: Preparation of (S)-5-((1-(6-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (33-d)

(S)-(1-(2-Aminopropyl)-6-fluoro-1H-indol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (33-c) (120 mg, purity 60%, 1 eq.) was dissolved in ethanol (3 ml), and triethylamine (49 mg, 3 eq.) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (63 mg, 1.2 eq.) were added, and the reaction was carried out at 60°C for 16 hours. After concentration under reduced pressure, the product was purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product (S)-5-((1-(6-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (33-d) (30 mg, yield 25.27%). ESI [M+H] ⁺ = 743.3.

Step 5: Preparation of (S)-5-((1-(6-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (33)

(S)-5-((1-(6-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (33-d) (30 mg, 1 eq.) was dissolved in 4 M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 5 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 10: 1) to give the target product (S)-5-((1-(6-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-indol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (33) (3.12 mg, yield 12.61%). ESI [M+H] ⁺ = 613.40. ¹ H NMR (400 MHz, CD ₃ OD) δ8.59 (s, 2H), 7.63 (s, 1H), 7.60 (dd, J=8.8, 5.2Hz, 1H), 7.39 (dd, J=9.8, 2.0Hz, 1H), 7.12 (s, 1H), 7.02-6.87 (m, 2H), 4.52 (d, J=12.0Hz, 2H), 4.43 (dd , J=14.6, 3.4Hz, 1H), 4.28-4.20 (m, 1H), 3.98 (t, J=5.0Hz, 3H), 3.75 (dd, J=11.0, 5.2Hz, 3H), 2.07-1.98 (m, 1H), 1.43 (d, J=6.4Hz, 3H).

Example 34: Preparation of (S)-5-((1-(2-chloro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (34)

Step 1: Preparation of 5-chloro-1H-pyrrole-3-carboxylic acid methyl ester (34-a)

1H-pyrrole-3-carboxylic acid methyl ester (100 mg, 1 eq.) was dissolved in tetrahydrofuran (2.5 ml), and pyridine (6 mg, 0.1 eq.) and 1-chloropyrrolidine-2,5-dione (107 mg, 1 eq.) were added at -70°C and reacted for 2 hours, then the temperature was raised to 30°C and stirred overnight. Water (10 ml) was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a flash column (petroleum ether: ethyl acetate = 4: 1) to obtain the target product (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-5-chloro-1H-pyrrole-3-carboxylic acid methyl ester (34-a) (120 mg, yield 94.10%). ESI [M+H] ⁺ = 160.1.

Step 2: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-5-chloro-1H-pyrrole-3-carboxylic acid methyl ester (34-b)

5-Chloro-1H-pyrrole-3-carboxylic acid methyl ester (34-a) (120 mg, 1 eq.) was dissolved in N, N-dimethylformamide (11 ml), cesium carbonate (735 mg, 3 eq.) was added, and then (S)-(1-iodopropane-2-yl)carbamic acid tert-butyl ester (322 mg, 1.5 eq.) was added, and stirred at 80°C overnight. Water (100 ml) was added to quench, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by Flash column (petroleum ether: ethyl acetate = 4: 1) to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-5-chloro-1H-pyrrole-3-carboxylic acid methyl ester (34-b) (163 mg, yield 68.42%). ESI [2M+H] ⁺ = 633.40.

Step 3: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-5-chloro-1H-pyrrole-3-carboxylic acid (34-c)

(S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-5-chloro-1H-pyrrole-3-carboxylic acid methyl ester (34-b) (163 mg, 1 eq.) was dissolved in methanol (2 ml), and water (2 ml) and sodium hydroxide (31 mg, 1.5 eq.) were added, and stirred overnight at 80°C in a sealed tube. The reaction solution was concentrated under reduced pressure to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-5-chloro-1H-pyrrole-3-carboxylic acid (34-c) (190 mg, purity 70%, yield 85.38%). ESI[2M+H] ⁺ =605.40.

Step 4: Preparation of tert-butyl-(1-(2-chloro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (34-d)

(S)-1-(2-((tert-Butyloxycarbonyl)amino)propyl)-5-chloro-1H-pyrrole-3-carboxylic acid (34-c) (190 mg, purity 70%, 1 eq.) was dissolved in dichloromethane (6 ml), and 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylhexafluorouronium (167 mg, 1 eq.) was added, followed by 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (122 mg, 1.2 eq.) and N,N-diisopropylethylamine (170 mg, 3 eq.), and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and then flash column (petroleum ether: ethyl acetate = 1: 1) was used to obtain the target product tert-butyl-(1-(2-chloro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (34-d) (150 mg, yield 66.05%). ESI [M+H] ⁺ = 517.30.

Step 5: Preparation of (S)-(1-(2-aminopropyl)-5-chloro-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (34-e)

Tert-butyl-(1-(2-chloro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (34-d) (150 mg, 1 eq.) was dissolved in dichloromethane (2 ml), trifluoroacetic acid (1.99 g, 60 eq.) was added, and stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the target product (S)-(1-(2-aminopropyl)-5-chloro-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (34-e) (170 mg, purity 70%, yield 98.39%). ESI [M+H] ⁺ = 417.30.

Step 6: Preparation of (S)-5-((1-(2-chloro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (34-f)

(S)-(1-(2-Aminopropyl)-5-chloro-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (34-e) (170 mg, purity 70%, 1 eq.) was dissolved in ethanol (4 ml), and triethylamine (87 mg, 3 eq.) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (113 mg, 1.2 eq.) were added, and the mixture was stirred at 60°C for 1 hour. The reaction solution was concentrated under reduced pressure and then flash column (petroleum ether: ethyl acetate = 0: 1) was used to obtain the target product (S)-5-((1-(2-chloro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (34-f) (90 mg, yield 44.45%). ESI [M+H] ⁺ = 709.40.

Step 7: Preparation of (S)-5-((1-(2-chloro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (34)

(S)-5-((1-(2-chloro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (34-f) (90 mg, 1 eq.) was dissolved in a hydrogen chloride ethyl acetate solution (3 ml, 4 mol/L) and stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and purified on a preparative plate (dichloromethane:methanol=10:1) to give the target product (S)-5-((1-(2-chloro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (34) (15.8 mg, yield 21.51%). ESI[M+H] ⁺ =579.50. ¹ H NMR (400MHz, CD ₃ OD) δ 8.59 (s, 2H), 7.41 (s, 1H), 7.16 (s, 1H), 6.28 (s, 1H), 4.40 (s, 1H), 4.23 (dd, J=14.5, 3.7Hz, 1H), 4.04 (dd, J =14.5, 9.9Hz, 1H), 3.99-3.86 (m, 4H), 3.76-3.69 (m, 3H), 1.55 (d, J = 6.9Hz, 1H), 1.38 (d, J = 6.5Hz, 3H).

Example 35: Preparation of (S)-6-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyrimidin-4(3H)-one (35)

Step 1: Preparation of tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (35-a)

(S)-1-(2-((tert-Butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (120 mg, 1 eq.) was dissolved in dichloromethane (6 ml), and 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylhexafluorouronium (170 mg, 1 eq.) was added, followed by 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (125 mg, 1.2 eq.) and N,N-diisopropylethylamine (173 mg, 3 eq.), and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and then flash column (petroleum ether: ethyl acetate = 1: 1) was used to obtain the target product tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (35-a) (187 mg, yield 86.66%). ESI [M+H] ⁺ = 483.3.

Step 2: Preparation of (S)-(1-(2-aminopropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (35-b)

Tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (35-a) (187 mg, 1 eq.) was dissolved in dichloromethane (2 ml), trifluoroacetic acid (2.65 g, 60 eq.) was added, and stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the target product (S)-(1-(2-aminopropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (35-b) (280 mg, purity 50%, yield 94.47%). ESI[M+H] ⁺ =383.3.

Step 3: Preparation of (S)-6-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-3-((2-(trimethylsilyl)ethoxy)methyl)pyrimidin-4(3H)-one (35-c)

(S)-(1-(2-Aminopropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (35-b) (140 mg, 1 eq.) was dissolved in ethanol (3 ml), and triethylamine (111 mg, 3 eq.) and 6-chloro-3-((2-(trimethylsilyl)ethoxy)methyl)pyrimidin-4(3H)-one (113 mg, 1.2 eq.) were added, and the mixture was stirred at 80°C for 24 hours. The reaction solution was concentrated under reduced pressure and then flash column (petroleum ether: ethyl acetate = 0: 1) was used to obtain the target product (S)-6-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-3-((2-(trimethylsilyl)ethoxy)methyl)pyrimidin-4(3H)-one (35-c) (17 mg, yield 15.31%). ESI [M+H] ⁺ = 607.5.

Step 4: Preparation of (S)-6-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyrimidin-4(3H)-one (35)

(S)-6-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-3-((2-(trimethylsilyl)ethoxy)methyl)pyrimidin-4(3H)-one (35-c) (17 mg, 1 eq.) was dissolved in a hydrogen chloride ethyl acetate solution (2 ml, 4 mol/L) and stirred at room temperature for 5 hours. The reaction solution was concentrated under reduced pressure and purified on a preparative plate (dichloromethane:methanol=10:1) to give the target product (S)-6-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyrimidin-4(3H)-one (35) (1.2 mg, yield 8.99%). ESI[M+H] ⁺ ＝477.20.

Example 36: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[3,2-b]pyridin-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (36)

Step 1: Preparation of (1H-pyrrolo[3,2-b]pyridin-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (36-a)

1H-pyrrolo[3,2-b]pyridine-3-carboxylic acid (100 mg, 1 eq.) was dissolved in dichloromethane (8 ml), 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylhexafluorouronium (234 mg, 1 eq.) was added, and then 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (172 mg, 1 eq.) and N,N-diisopropylethylamine (239 mg, 3 eq.) were added, and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and flash column (petroleum ether: ethyl acetate = 1:1) was used to obtain the target product (1H-pyrrolo[3,2-b]pyridin-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (36-a) (77 mg, yield 33.18%).

Step 2: Preparation of tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[3,2-b]pyridin-1-yl)propan-2-yl)carbamate (36-b)

(1H-pyrrolo[3,2-b]pyridin-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (36-a) (77 mg, 1 eq.) was dissolved in N,N-dimethylformamide (4 ml), cesium carbonate (200 mg, 3 eq.) was added, and then (S)-tert-butyl(1-iodopropane-2-yl)carbamate (76 mg, 1.3 eq.) was added, and the mixture was stirred at 80°C overnight. Water (30 ml) was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[3,2-b]pyridin-1-yl)propane-2-yl)carbamate (36-b) (92 mg, yield 84.28%). ESI [M+H] ⁺ = 534.4.

Step 3: Preparation of (S)-(1-(2-aminopropyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (36-c)

Tert-butyl-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[3,2-b]pyridin-1-yl)propan-2-yl)carbamate (36-b) (92 mg, 1 eq.) was dissolved in dichloromethane (1 ml), trifluoroacetic acid (1 ml) was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the target product (S)-(1-(2-aminopropyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (36-c) (100 mg, purity 60%, yield 80.28%). ESI[M+H] ⁺ =434.3.

Step 4: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[3,2-b]pyridin-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (36-d)

(S)-(1-(2-Aminopropyl)-1H-pyrrolo[3,2-b]pyridin-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (36-c) (100 mg, purity 60%, 1 eq.) was dissolved in ethanol (2 ml), and triethylamine (42 mg, 3 eq.) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (69 mg, 1.5 eq.) were added, and the mixture was stirred at 60°C for 16 hours. The reaction solution was concentrated under reduced pressure and then purified by flash column (petroleum ether: ethyl acetate = 0:1) to obtain the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[3,2-b]pyridin-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (36-d) (80 mg, yield 79.63%).

Step 5: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[3,2-b]pyridin-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (36)

(S)-4-(Trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[3,2-b]pyridin-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (36-d) (80 mg, 1 eq.) was dissolved in hydrogen chloride ethyl acetate solution (3 ml, 4 mol/L) and stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and purified on a preparative plate (dichloromethane:methanol=10:1) to give the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrolo[3,2-b]pyridin-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (36) (6 mg, yield 9.14%). ESI[M+H] ⁺ =596.30. ¹ H NMR (400MHz, CD ₃ OD) δ 8.56 (d, J = 13.6Hz, 2H), 8.41 (d, J = 4.4Hz, 1H), 8.09 (d, J = 8.3Hz, 1H), 7.97 (s, 1H), 7.30 (dd, J = 8.4, 4.7Hz, 1H), 7.10 (s, 1H), 4.55-4.50 (m, 1H), 4.34 (dd, J=15.1, 10.8Hz, 1H), 3.64 (d, J=10.7Hz, 4H), 3.57-3.44 (m, 1H), 3.30 (dt, J=3.2, 1.6Hz, 4H), 1.27 (s, 3H).

Example 37: Preparation of (S)-6-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)nicotinonitrile (37)

Step 1: Preparation of (S)-6-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)nicotinonitrile (37-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (50 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (41 mg, 1 eq.), N,N-diisopropylethylamine (42 mg, 3 eq.) and 6-(piperazin-1-yl)nicotinonitrile (20 mg, 1 eq.) were added and reacted at room temperature for 2 hours. After concentration under reduced pressure, the product was purified by preparative plate (petroleum ether: ethyl acetate = 0: 1) to obtain the target product (S)-6-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)nicotinonitrile (37-a) (17 mg, yield 24.82%). ESI [M+H] ⁺ = 631.4.

Step 2: Preparation of (S)-6-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)nicotinonitrile (37)

(S)-6-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)nicotinonitrile (37-a) (17 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 3 hours. After concentration under reduced pressure, the mixture was purified by preparative plate (dichloromethane:methanol=10:1) to obtain the target product (S)-6-(4-(1-(2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-1-yl)nicotinonitrile (37) (3 mg, yield 22.24%). ESI[M+H] ⁺ =501.30. ¹ H NMR (600MHz, CD ₃ OD) δ 8.42 (s, 1H), 7.76 (dd, J = 9.1, 2.3Hz, 1H), 7.37 (s, 1H), 7.12 (s, 1H), 6.88 (d, J = 9.1Hz, 1H), 6.77 (t, J = 2.5Hz, 1 H), 6.30-6.28 (m, 1H), 4.43-4.30 (m, 2H), 4.20 (dd, J=14.2, 3.7Hz, 1H), 3.94 (dd, J=14.2, 10.0Hz, 1H), 3.76 (dd, J=14.5, 7.5Hz, 7H), 1.35 (d, J=6.5Hz, 3H).

Example 38: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (38)

Step 1: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (38-a)

1H-pyrrole-3-carboxylic acid methyl ester (245 mg, 1.96 mmol) was dissolved in N,N-dimethylformamide (10 ml), cesium carbonate (1.92 g, 5.89 mmol) was added, and the mixture was stirred at room temperature for 30 minutes, and then (S)-(1-iodopropane-2-yl)carbamic acid tert-butyl ester (840 mg, 2.95 mmol) was added, and the mixture was stirred at 80°C overnight. Water (10 ml) was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by Flash column (dichloromethane: methanol = 25: 1) to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (38-a) (415 mg, yield 75.0%). ESI[M-56+H] ⁺ =227.1, ESI[2M+H] ⁺ =565.6, ESI[2M+Na] ⁺ =587.4

Step 2: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (38-b)

(S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (38-a) (200 mg, 0.709 mmol) was dissolved in methanol (10 ml), and water (3 ml) and sodium hydroxide (57 mg, 1.43 mmol) were added, and the mixture was stirred at 80°C in a sealed tube overnight. The pH of the reaction solution was adjusted to 7 with 1N hydrochloric acid, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (38-b) (180 mg, yield 94.7%). ESI[M-56+H] ⁺ =213.0, ESI[2M+H] ⁺ =537.4, ESI[2M+Na] ⁺ =559.4

Step 3: Preparation of tert-butyl (S)-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (38-c)

(S)-1-(2-((tert-Butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (38-b) (18 mg, 0.067 mmol) was dissolved in dichloromethane (3 ml), 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylhexafluorouronium (38 mg, 0.10 mmol) was added, and stirred at room temperature for 15 minutes. Then 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (18 mg, 0.067 mmol) and N,N-diisopropylethylamine (29 ul, 0.168 mmol) were added and stirred at room temperature for 3 hours. Saturated ammonium chloride solution (3 ml) was added to quench the reaction, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product, tert-butyl (S)-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (38-c) (30 mg, yield 93.8%). ESI [M+H] ⁺ = 483.3, ESI [2M+H] + = 965.6, ESI [M-56+H] ⁺ = 427.3.

Step 4: Preparation of (S)-(1-(2-aminopropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone trifluoroacetate (38-d)

Tert-butyl (S)-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (38-c) (30 mg, 0.062 mmol) was dissolved in dichloromethane (3 ml), trifluoroacetic acid (0.5 ml) was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the target product (S)-(1-(2-aminopropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone trifluoroacetate (38-d) (35 mg, yield 113.3%).

Step 5: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (38-e)

(S)-(1-(2-Aminopropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone trifluoroacetate (38-d) (30 mg, 0.060 mmol) was dissolved in ethanol (3 ml), and triethylamine (2 lul, 0.15 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (20 mg, 0.060 mmol) were added, and stirred at 60 ° C for 4.5 hours. The reaction solution was concentrated, and the residue was dissolved in dichloromethane, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (38-e) (28 mg, yield 68.6%). ESI[M+H] ⁺ = 675.4, ESI[M-28] ⁺ = 647.3.

Step 6: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (38)

(S)-4-(Trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (38-e) (28 mg, 0.042 mmol) was dissolved in hydrogen chloride ethyl acetate solution (3 ml, 4 mol/L) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and purified on a preparative plate (dichloromethane:methanol=20:1) to give the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (38) (13 mg, yield 57.5%). ESI[M+H] ⁺ =545.40, ¹ H NMR (400MHz, CDCl ₃ ) δ10.51 (s, 1H), 8.51 (s, 2H), 7.11 (d, J = 5.2Hz, 2H), 6.57 (t, J = 2.3Hz, 1H), 6.30 (s, 1H), 5.33-5.24 (m, 1H), 4 .11 (dd, J=19.9, 6.3Hz, 2H), 3.93 (s, 4H), 3.78 (d, J=5.2Hz, 4H), 3.64 (s, 1H), 1.42-1.37 (m, 3H).

Example 39: Preparation of 5-((S)-1-(3-((R)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (39)

Step 1: Preparation of 5-((S)-1-(3-((R)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (39-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (32 mg, 1 eq.), N,N-diisopropylethylamine (34 mg, 3 eq.) and (R)-2-(2-methylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (26 mg, 1.2 eq.) were added and reacted at room temperature for 2 hours. After concentration under reduced pressure, the product was purified by preparative plate (petroleum ether: ethyl acetate = 0: 1) to obtain the target product 5-((S)-1-(3-((R)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (39-a) (40 mg, yield 66%). ESI [M+H] ⁺ = 689.4.

Step 2: Preparation of 5-((S)-1-(3-((R)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (39)

5-((S)-1-(3-((R)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (39-a) (40 mg, 1 eq.) was dissolved in 4 M hydrochloric acid ethyl acetate solution (3 ml) and reacted at room temperature for 1 hour. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to give the target product 5-((S)-1-(3-((R)-3-methyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (39) (23.7 mg, yield 73.07%). ESI[M+H] ⁺ =559.3. ¹ H NMR (400MHz, CD ₃ OD) δ 8.58 (s, 2H), 7.40 (s, 1H), 7.14 (s, 1H), 6.79-6.75 (m, 1H), 6.31-6.28 (m, 1H), 4.57 (dd, J = 10.4, 3.1Hz, 2H), 4 .41-4.25 (m, 4H), 4.20 (dd, J=14.1, 3.7Hz, 2H), 3.96 (dd, J=14.1, 9.9Hz, 2H), 3.20 (s, 1H), 1.34 (d, J=6.5Hz, 3H), 1.18 (d, J=6.7Hz, 3H).

Example 40: Preparation of ethyl 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-2-yl) (40)

Step 1: Preparation of tert-butyl (S)-3-(cyanomethyl)-4-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazine-1-carboxylate (40-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (25 mg, 1 eq.) was dissolved in N,N-dimethylformamide (1.5 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (20 mg, 1 eq.), N,N-diisopropylethylamine (20 mg, 3 eq.) and (S)-3-(cyanomethyl)piperazine-1-carboxylic acid tert-butyl ester (11 mg, 1.2 eq.) were added and reacted at 100°C overnight. Water (20 ml) was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure and then purified by flash column (petroleum ether: ethyl acetate = 1: 2) to obtain the target product tert-butyl (S)-3-(cyanomethyl)-4-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazine-1-carboxylate (40-a) (21 mg, yield 75.31%). ESI [M+H] ⁺ = 668.4.

Step 2: Preparation of methyl 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-2-yl)acetate (40-b)

Tert-butyl (S)-3-(cyanomethyl)-4-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazine-1-carboxylate (40-a) (21 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 1 hour. After concentration under reduced pressure, the target product, methyl 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-2-yl)acetate (40-b) (12 mg, yield 81.11%) was obtained. ESI[M+H] ⁺ ＝471.3.

Step 3: Preparation of ethyl acetate 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-2-yl) (40)

Methyl 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazine-2-yl)acetate (40-b) (12 mg, 1 eq.) was dissolved in N-methylpyrrolidone (1.5 ml), and 2-chloro-5-(trifluoromethyl)pyrimidine (6 mg, 1.2 eq.) and potassium carbonate (11 mg, 3 eq.) were added, and the reaction was carried out at 80°C for 1 hour. Water (5 ml) was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and then purified by preparative HPLC and freeze-dried to obtain the target product, ethyl acetate 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-2-yl)(40) (1 mg, yield 6.22%). ESI[M+H] ⁺ =631.4.

Example 41: Preparation of 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-2-yl)acetamide (41)

Step 1: Preparation of 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-2-yl)acetamide (41-a)

Tert-butyl (S)-3-(cyanomethyl)-4-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazine-1-carboxylate (21 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 1 hour. After concentration under reduced pressure, the target product 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-2-yl)acetamide (41-a) (12 mg, yield 81.11%) was obtained.

Step 2: Preparation of 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-2-yl)acetamide (41)

2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)piperazin-2-yl)acetamide (41-a) (12 mg, 1 eq.) was dissolved in N-methylpyrrolidone (1.5 ml), and 2-chloro-5-(trifluoromethyl)pyrimidine (6 mg, 1.2 eq.) and potassium carbonate (11 mg, 3 eq.) were added, and the reaction was carried out at 80°C for 1 hour. Water (5 ml) was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and purified by preparative HPLC and freeze-dried to obtain the target product 2-((S)-1-(1-((S)-2-((6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carbonyl)-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-2-yl)acetamide (41) (0.93 mg, yield 6.06%). ESI [M+H] + = 602.4.

Example 42: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (42)

Step 1: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (42-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 1 eq.), N,N-diisopropylethylamine (34 mg, 3 eq.) and 2-(piperazin-1-yl)-5-(trifluoromethyl)pyrazine (24 mg, 1.eq.) were added, and the mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and then purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (42-a) (54 mg, yield 92.92%). ESI [M+H] ⁺ = 675.4.

Step 2: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (42)

(S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (42-a) (54 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (3 ml) and reacted at room temperature for 16 hours. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to give the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (42) (22.5 mg, yield 51.64%). ESI[M+H] ⁺ =545.4. ¹ H NMR (400MHz, CD ₃ OD) δ 8.40 (s, 1H), 8.30 (s, 1H), 7.37 (s, 1H), 7.13 (t, J = 1.6Hz, 1H), 6.77 (t, J = 2.4Hz, 1H), 6.31-6.27 (m, 1H), 4.4 0-4.31 (m, 1H), 4.20 (dd, J=14.1, 3.7Hz, 1H), 3.94 (dd, J=14.1, 10.0Hz, 1H), 3.79 (s, 8H), 1.35 (d, J=6.5Hz, 3H).

Example 43: Preparation of 5-((S)-1-(3-((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (43)

Step 1: Preparation of 5-((S)-1-(3-((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (43-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 1 eq.), N,N-diisopropylethylamine (34 mg, 3 eq.) and 2-((3R,5S)-3,5-dimethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (23 mg, 1 eq.) were added, and the reaction was carried out at 80°C for 16 hours. The reaction solution was quenched by adding water (10 ml), extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product 5-((S)-1-(3-((2R, 6S)-2, 6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (43-a) (21 mg, yield 33.81%). ESI [M+H] + = 703.5.

Step 2: Preparation of 5-((S)-1-(3-((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (43)

(5-((S)-1-(3-((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (43-a) (21 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 3 hours. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to give the target product 5-((S)-1-(3-((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (43) (1.5 mg, yield 51.64%). ESI[M+H]+=573.4.1H NMR (600MHz, CD3OD) δ8.58 (s, 2H), 7.38 (s, 1H), 7.10 (s, 1H), 6.77 (t, J=2.4Hz, 1H), 6.32-6.29 (m, 1H), 4.72 (d, J=13.4Hz, 3H), 4.41-4. 29 (m, 2H), 4.20 (dd, J=14.2, 3.7Hz, 1H), 3.96 (dd, J=14.2, 9.9Hz, 1H), 3.23 (d, J=12.5Hz, 2H), 1.35 (d, J=6.5Hz, 3H), 1.28 (t, J=6.8Hz, 6H).

Example 44: Preparation of 5-((S)-1-(3-((R)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (44)

Step 1: Preparation of 5-((S)-1-(3-((R)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (44-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 1 eq.), N,N-diisopropylethylamine (34 mg, 3 eq.) and (R)-2-(3-ethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (27 mg, 1.2 eq.) were added and reacted at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and then purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product 5-((S)-1-(3-((R)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (44-a) (52 mg, yield 85.59%). ESI [M+H] + = 703.4.

Step 2: Preparation of 5-((S)-1-(3-((R)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (44)

5-((S)-1-(3-((R)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (44-a) (52 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (3 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the product was purified by preparative HPLC and freeze-dried to give the target product 5-((S)-1-(3-((R)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (44) (20.9 mg, yield 49.34%). ESI[M+H] ⁺ =573.4. ¹ H NMR (400 MHz, CD ₃ OD) δ8.57 (s, 2H), 7.39 (s, 1H), 7.12 (s, 1H), 6.76 (t, J=2.4Hz, 1H), 6.26 (s, 1H), 4.36 (s, 2H), 4.19 (dd, J=14.1, 3.6Hz, 1H), 3.95 (dd, J=14.1, 9.9Hz, 1H) , 3.30 (d, J=1.5Hz, 3H), 3.19 (dd, J=13.6, 3.3Hz, 1H), 3.02 (td, J=12.7, 3.3Hz, 1H), 1.70-1.48 (m, 3H), 1.34 (d, J=6.5Hz, 3H), 0.85 (s, 3H).

Example 45: Preparation of 5-((S)-1-(3-((R)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (45)

Step 1: Preparation of 5-((S)-1-(3-((R)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (45-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 1 eq.), N,N-diisopropylethylamine (34 mg, 3 eq.) and (R)-2-(3-isopropylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (24 mg, 1 eq.) were added, and the reaction was carried out at 80°C for 16 hours. The reaction solution was quenched by adding water (10 ml), extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product 5-((S)-1-(3-((R)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (45-a) (42 mg, yield 66.97%). ESI [M+H] + = 717.4.

Step 2: Preparation of 5-((S)-1-(3-((R)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (45)

5-((S)-1-(3-((R)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (45-a) (42 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (3 ml) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to give the target product 5-((S)-1-(3-((R)-2-isopropyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (45) (13.9 mg, yield 40.45%). ESI[M+H] ⁺ =587.4. ¹ H NMR (400MHz, CD ₃ OD) δ 8.56 (s, 2H), 7.40 (s, 1H), 7.11 (s, 1H), 6.78 (s, 1H), 6.31-6.24 (m, 1H), 4.38 (s, 2H), 4.23-4.08 (m, 2H), 4. 09-3.85 (m, 2H), 3.22-3.02 (m, 2H), 2.96 (d, J=11.9Hz, 2H), 2.01-1.91 (m, 1H), 1.34 (d, J=6.4Hz, 3H), 1.09 (d, J=5.6Hz, 3H), 0.84 (d, J=5.8Hz, 3H).

Example 46: Preparation of 5-((S)-1-(3-((S)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (46)

Step 1: Preparation of 5-((S)-1-(3-((S)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (46-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 1 eq.) was dissolved in dichloromethane (2 ml), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 1 eq.), N,N-diisopropylethylamine (34 mg, 3 eq.) and (S)-2-(3-ethylpiperazin-1-yl)-5-(trifluoromethyl)pyrimidine (23 mg, 1 eq.) were added and reacted at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and then purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product 5-((S)-1-(3-((S)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (46-a) (26 mg, yield 42.59%). ESI [M+H] ⁺ = 703.5.

Step 2: Preparation of 5-((S)-1-(3-((S)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (46)

5-((S)-1-(3-((S)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (46-a) (26 mg, 1 eq.) was dissolved in 4M hydrochloric acid ethyl acetate solution (2 ml) and reacted at room temperature for 3 hours. After concentration under reduced pressure, the product was purified by preparative HPLC and freeze-dried to give the target product 5-((S)-1-(3-((S)-2-ethyl-4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (46) (9.3 mg, yield 43.91%). ESI[M+H] ⁺ =573.4. ¹ H NMR (400 MHz, CD ₃ OD) δ8.57 (s, 2H), 7.38 (s, 1H), 7.10 (s, 1H), 6.77 (s, 1H), 6.27-6.23 (m, 1H), 4.36 (s, 2H), 4.20 (dd, J=14.1, 3.6Hz, 1H), 3.94 (dd, J=14.0, 10.1Hz, 1H), 3.34 (s, 1H), 3.18 (d, J = 13.4Hz, 2H), 3.03 (t, J = 12.7Hz, 2H), 1.60 (ddd, J = 20.9, 13.8, 6.7Hz, 3H), 1.34 (d, J = 6.5Hz, 3H), 0.86 (s, 3H).

Example 47: Preparation of (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl-2,2,3,3,5,5,6,6-d8)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (47)

Step 1: Preparation of (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl-2,2,3,3,5,5,6,6-d8)-1H-pyrrol-1-yl)propane-2-yl)amino)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (47-a)

(S)-1-(2-(6-oxo-5-(trifluoromethyl)-1-(2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (40 mg, 0.086 mmol) was dissolved in dichloromethane (2 mL), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 0.086 mmol), N,N-diisopropylethylamine (34 mg, 0.258 mmol) and 2-(piperazin-1-yl-2,2,3,3,5,5,6,6-d8)-5-(trifluoromethyl)pyrimidine hydrochloride (24 mg, 0.104 mmol) were added and reacted at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and then purified by Flash column (petroleum ether: ethyl acetate = 0: 1) to obtain the target product (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl-2,2,3,3,5,5,6,6-d8)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (47-a) (40 mg, yield 67.45%). ESI [M+H] ⁺ = 683.5

Step 2: Preparation of (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl-2,2,3,3,5,5,6,6-d8)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (47)

(S)-4-(Trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl-2,2,3,3,5,5,6,6-d8)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazine-3(2H)-one (47-a) (40 mg, 0.058 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (1 mL) and reacted at room temperature for 4 hours. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to give the target product (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl-2,2,3,3,5,5,6,6-d8)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (47) (10.1 mg, yield 31.20%). ESI[M+H] ⁺ =553.3. ¹ H NMR (400MHz, CD ₃ OD) δ 8.59 (s, 2H), 7.38 (s, 1H), 7.15 (s, 1H), 6.78-6.74 (m, 1H), 6.30-6.27 (m, 1H), 4.76 (s, 3H), 4.37 (s, 3H), 4.21 (dd, J=14.1, 3.6Hz, 2H), 3.95 (dd, J=14.1, 10.0Hz, 3H), 1.35 (d, J=6.5Hz, 3H).

Example 48: Preparation of (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (48)

Step 1: Preparation of tert-butyl (S)-methyl (1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (48-a)

(S)-1-(2-(tert-Butyloxycarbonyl)(methyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (70 mg, 0.248 mmol) was dissolved in dichloromethane (3 mL), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (94 mg, 0.248 mmol), N,N-diisopropylethylamine (96 mg, 0.744 mmol) and 1-(5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (66 mg, 0.248 mmol) were added and reacted at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and purified by Flash column (ethyl acetate: petroleum ether = 0% to 70%) to obtain the target product tert-butyl (S)-methyl (1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (48-a) (86 mg, yield 70%). ESI [M+H] ⁺ = 496.4

Step 2: Preparation of (S)-(1-(2-(methylamino)propyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-yl)methanone (48-b)

Tert-butyl (S)-methyl (1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (48-a) (86 mg, 0.174 mmol) was dissolved in 4M hydrochloric acid-dioxane (2 mL) and reacted at room temperature for 1 hour. After concentration under reduced pressure, the target product (S)-(1-(2-(methylamino)propyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-yl)methanone (48-b) (60 mg, yield 86.43%) was obtained. ESI [M+H] ⁺ = 396.3

Step 3: Preparation of (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (48-c)

(S)-(1-(2-(Methylamino)propyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-yl)methanone (48-b) (60 mg, 0.152 mmol) was dissolved in ethanol (2 mL), and triethylamine (46 mg, 0.456 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazine-3(2H)-one (60 mg, 0.182 mmol) were added, and the reaction was carried out at 60°C for 4 hours. After concentration under reduced pressure, the mixture was purified by flash column (ethyl acetate: petroleum ether = 0% to 70%) to obtain the target product (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (48-c) (51 mg, yield 48.87%). ESI [M+H] ⁺ = 688.3

Step 4: Preparation of (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (48)

(S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (48-c) (51 mg, 0.074 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (2.5 mL) and reacted at room temperature for 16 hours. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to obtain the target product (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (48) (27 mg, yield 65.31%). ESI[M+H] ⁺ =558.2. ¹ H NMR (600MHz, CD ₃ OD) δ 8.36 (s, 1H), 7.77 (d, J = 8.9Hz, 1H), 7.36 (s, 1H), 7.14 (s, 1H), 6.95 (d, J = 9.1Hz, 1H), 6.74 (s, 1H), 6.26 (s, 1H ), 4.30 (s, 1H), 4.23-4.16 (m, 1H), 4.04 (d, J=14.4Hz, 1H), 3.83 (dd, J=69.0, 24.2Hz, 8H), 3.12-2.99 (m, 3H), 1.37 (dd, J=34.7, 6.7Hz, 3H).

Example 49: Preparation of (S)-5-(1-(3-(4-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (49)

Step 1: Preparation of (S)-5-(1-(3-(4-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (49-a)

(S)-1-(2-(6-oxo-5-(trifluoromethyl)-1-(2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxamide (40 mg, 0.087 mmol) was dissolved in dichloromethane (6 mL), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (33 mg, 0.087 mmol), N,N-diisopropylethylamine (34 mg, 0.261 mmol) and 1-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine hydrochloride (30 mg, 0.104 mmol) were added and reacted at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and then purified by Flash column (ethyl acetate: petroleum ether = 0% to 70%) to obtain the target product (S)-5-(1-(3-(4-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (49-a) (20 mg, yield 33.29%). ESI [M+H] ⁺ = 692.3

Step 2: Preparation of (S)-5-(1-(3-(4-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (49)

(S)-5-(1-(3-(4-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (49-a) (20 mg, 0.029 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (2 mL) and reacted at room temperature for 4 hours. After concentration under reduced pressure, the mixture was purified by preparative HPLC and freeze-dried to obtain the target product (S)-5-(1-(3-(4-(3-fluoro-5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (49) (6 mg, yield 36.96%). ESI[M+H] ⁺ =562.4. ¹ H NMR (400MHz, CD ₃ OD) δ 8.27 (s, 1H), 7.67 (dd, J = 13.3, 1.3Hz, 1H), 7.38 (s, 1H), 7.13 (s, 1H), 6.79-6.73 (m, 1H), 6.29 (s, 1H), 4.36 (s , 1H), 4.20 (dd, J=14.1, 3.6Hz, 1H), 3.95 (dd, J=14.1, 9.9Hz, 1H), 3.78 (dd, J=11.5, 8.6Hz, 4H), 3.74-3.60 (m, 4H), 1.33 (t, J=12.0Hz, 3H).

Example 50: Preparation of (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)oxy)pyridazin-3(2H)-one (50)

Step 1: Preparation of (1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (50-a)

To a dichloromethane solution (10 mL) of 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (523 mg, 0.023 mmol) were added 1H-pyrrole-3-carboxylic acid (250 mg, 0.023 mmol), 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (1.3 g, 0.034 mmol), and N,N-diisopropylethylamine (1.2 mL, 0.068 mmol) at room temperature. After stirring at room temperature for 4 hours, the reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (dichloromethane: methanol = 100/1) to give (1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidine-2-yl)piperazine-1-yl)methanone (50-a) (559 mg, yield 76.37%). ESI[M+H] ⁺ ＝326.2.

Step 2: Preparation of (S)-(1-(2-hydroxypropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (50-b)

At room temperature, (R)-2-methyloxirane (299 mg, 0.052 mmol) and potassium carbonate (474.3 mg, 0.034 mmol) were added to a solution of (1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (50-a) (559 mg, 0.017 mmol) in N,N-dimethylformamide (10 mL). The mixture was stirred overnight at 100 degrees Celsius. The reaction solution was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (solvent: ethyl acetate) to give (S)-(1-(2-hydroxypropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (50-b) (180 mg, yield 27.3%). ESI[M+H] ⁺ =384.3.

Step 3: Preparation of (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)oxy)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (50-c)

To a solution of (S)-(1-(2-hydroxypropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (50-b) (180 mg, 0.470 mmol) in acetonitrile (10 mL) were added 5-chloro-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (154.4 mg, 0.470 mmol), cesium carbonate (306 mg, 0.470 mmol) and 1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (50-b) (180 mg, 0.470 mmol) at room temperature. 939mmol), stirred at room temperature overnight, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (solvent: ethyl acetate) to obtain (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)oxy)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (50-c) (60mg, yield 34%). ESI[M+H] ⁺ =676.3.

Step 4: Preparation of (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)oxy)pyridazin-3(2H)-one (50)

At room temperature, (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)oxy)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (50-c) (60 mg, 0.088 mmol) was dissolved in 4M hydrochloric acid ethyl acetate (4 mL) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and the crude product was purified by preparative HPLC to give (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)oxy)pyridazin-3(2H)-one (50) (5.1 mg, yield 12.1%). ESI[M+H] ⁺ ＝546.2.

Example 51: Preparation of (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)oxy)pyridazin-3(2H)-one (51)

Step 1: Preparation of (1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-yl)methanone (51-a)

To a solution of 1-(5-(trifluoromethyl)pyridin-2-yl)piperazine (70 mg, 0.303 mmol) in dichloromethane (10 mL) were added 1H-pyrrole-3-carboxylic acid (33.6 mg, 0.303 mmol), 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (172.6 mg, 0.481 mmol), N,N-diisopropylethylamine (158 u L, 0.962mmol), stirred at room temperature for 4 hours, water (10mL) was added to quench the reaction, extracted with ethyl acetate, the organic phase was washed with brine and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (dichloromethane: methanol = 100/1) to obtain (1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-yl)methanone (51-a) (69mg, yield 70.2%). ESI [M+H] ⁺ = 325.2.

Step 2: Preparation of (S)-(1-(2-hydroxypropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-yl)methanone (51-b)

To a solution of (1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-yl)methanone (51-a) (69 mg, 0.213 mmol) in N,N-dimethylformamide (2 mL) were added (R)-2-methyloxirane (31.7 mg, 0.638 mmol) and potassium carbonate (58.72 mg, 0.426 mmol) at room temperature, and the mixture was stirred at 100 degrees Celsius overnight. Water (10 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with brine and filtered. The filtrate was concentrated under reduced pressure and the crude product was purified by silica gel chromatography (ethyl acetate) to give (S)-(1-(2-hydroxypropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-yl)methanone (51-b) (50 mg, yield 61.46%). ESI[M+H] ⁺ ＝383.3.

Step 3: Preparation of (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)oxy)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (51-c)

To a solution of (S)-(1-(2-hydroxypropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazin-1-yl)methanone (51-b) (50 mg, 0.1.1 mmol) in acetonitrile (2 mL) were added 5-chloro-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (43 mg, 0.131 mmol), cesium carbonate (85.2 mg, 0.1.1 mmol) and 1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyridin-2-yl)piperazin-1-yl)methanone (51-b) (50 mg, 0.1.1 mmol) at room temperature. 262mmol), stirred at room temperature overnight, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (ethyl acetate) to obtain (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)oxy)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (51-c) (30mg, yield 34%). ESI[M+H] ⁺ =675.3.

Step 4: Preparation of (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)oxy)pyridazin-3(2H)-one (51)

At room temperature, (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)oxy)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (51-c) (30 mg, 0.045 mmol) was dissolved in 4M hydrochloric acid ethyl acetate (4 mL) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and the crude product was purified by preparative high performance liquid chromatography to give (S)-4-(trifluoromethyl)-5-(1-(3-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)oxy)pyridazin-3(2H)-one (51) (4.8 mg, yield 19.8%). ESI [M+H]+ = 545.2. ¹ H NMR (400MHz, CD ₃ OD) δ 8.37 (s, 1H), 7.81 (s, 1H), 7.74 (d, J = 2.3Hz, 1H), 7.18 (s, 1H), 6.91 (d, J = 9.0Hz, 1H), 6.82-6.77 (m, 1H), 6.33 ( d, J=1.8Hz, 1H), 4.33 (dd, J=14.8, 2.6Hz, 2H), 4.18-4.13 (m, 1H), 3.81 (s, 4H), 3.73 (d, J=4.2Hz, 4H), 1.43 (d, J=6.2Hz, 3H).

Example 52: Preparation of 5-((S)-1-(3-(2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (52)

Step 1: Preparation of tert-butyl ((S)-1-(3-(2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (52-a)

To a solution of ((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazin-1-yl)(1H-pyrrol-3-yl)methanone (175 mg, 0.495 mmol) in N,N-dimethylformamide (5 mL) was added cesium carbonate (434.5 mg, 1.276 mmol) at room temperature. After stirring at room temperature for 30 minutes, tert-butyl (S)-(1-iodopropane-2-yl)carbamate (253.4 mg, 0.991 mmol) was added. l), reacted at 100 degrees Celsius overnight, then water (10 mL) was added to quench the reaction, extracted with ethyl acetate, the organic phase was washed with brine and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (ethyl acetate) to obtain tert-butyl ((S)-1-(3-(2R, 6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)carbamate (52-a) (68 mg, yield 30%). ESI [M+H] ⁺ = 511.3.

Step 2: Preparation of (1-((S)-2-aminopropyl)-1H-pyrrol-3-yl)((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazin-1-yl)methanone (52-b)

At room temperature, tert-butyl ((S)-1-(3-(2R, 6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (52-a) (68 mg, 0.133 mmol) was dissolved in ethyl acetate (5 mL), and a 4M solution of dioxane hydrochloride (5 mL) was added. After stirring at room temperature for 2 hours, the mixture was filtered to obtain (1-((S)-2-aminopropyl)-1H-pyrrol-3-yl)((2R, 6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazin-1-yl)methanone (52-b) (50 mg, yield 91.4%). ESI [M+H] ⁺ = 411.2.

Step 3: Preparation of 5-((S)-1-(3-(2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyrazin-3(2H)-one (52-c)

To a solution (5 mL) of (1-((S)-2-aminopropyl)-1H-pyrrol-3-yl)((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazin-1-yl)methanone (52-b) (50 mg, 0.122 mmol) in ethanol was added 5-chloro-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (52-b) (40 mg, 0.122 mmol), triethylamine (17 uL, 0.365 mL) and 1-((S)-2-aminopropyl)-1H-pyrrol-3-yl)((2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazin-1-yl)methanone (52-b) (50 mg, 0.122 mmol) in ethanol at room temperature (5 mL). mol), stirred at 60 degrees Celsius for 4 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography (dichloromethane: methanol = 100/1) to obtain 5-((S)-1-(3-(2R, 6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyrazin-3(2H)-one (52-c) (19 mg, yield 22%). ESI[M+H] ⁺ =703.3.

Step 4: Preparation of 5-((S)-1-(3-(2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (52)

5-((S)-1-(3-(2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyrazin-3(2H)-one (52-c) (19 mg, 0.027 mmol) was dissolved in 4 M hydrochloric acid ethyl ester (3 M) at room temperature. mL), stirred at room temperature for 2 hours, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative high performance liquid chromatography to give 5-((S)-1-(3-(2R,6S)-2,6-dimethyl-4-(5-(trifluoromethyl)pyrazin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (52) (8.1 mg, yield 52.3%). ESI[M+H] ⁺ =573.4. ¹ H NMR (400 MHz, CD ₃ OD) δ 8.37 (d, J = 18.6Hz, 2H), 7.36 (s, 1H), 7.09 (s, 1H), 6.78 (s, 1H), 6.31 (d, J = 12.2Hz, 1H), 4.66 (s, 2H), 4.37 (d, J = 7.8Hz, 2H), 4.35 (s, 1H), 4.21 (dd, J=14.1, 3.3Hz, 1H), 4.09 (dd, J=14.2, 7.1Hz, 1H), 3.95 (dd, J=14.0, 10.1Hz, 1H), 2.02 (d, J=9.2Hz, 1H), 1.33 (dt, J=20.6, 10.2Hz, 9H).

Example 53: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (53)

Step 1: Preparation of 4-(trifluoromethyl)-5-(2S)-1-(3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (53-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (130 mg, 0.283 mmol) was dissolved in dichloromethane (10 mL), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (161 mg, 0.424 mmol), N,N-diisopropylethylamine (128 ul, 0.736 mmol) and 3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane hydrochloride (83 mg, 0.283 mmol) were added and reacted at room temperature for 2 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product 4-(trifluoromethyl)-5-(2S)-1-(3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (53-a) (74 mg, yield 37.4%). ESI [M+H] ⁺ = 700.5

Step 2: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (53)

4-(Trifluoromethyl)-5-(2S)-1-(3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (53-a) (74 mg, 0.106 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (5 mL) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the mixture was purified by preparative plate (dichloromethane:methanol=15:1) to give the target product 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyridin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (53) (34 mg, yield 56.7%). ESI[M+H] ⁺ =570.4. ¹ H NMR (600 MHz, CD ₃ OD) δ8.36 (s, 1H), 7.75 (d, J=8.2Hz, 1H), 7.35 (s, 1H), 7.18 (s, 1H), 6.85 (d, J=8.5Hz, 1H), 6.80 (s, 1H), 6.37 (s, 1H), 4.38 (s, 1H), 4.28-4.08 (m, 3H), 4.01-3.91 (m, 1H), 3.31 (s, 2H), 3.14 (dd, J=19.4, 12.7Hz, 2H), 1.93 (s, 2H), 1.79 (d, J=7.5Hz, 2H), 1.37 (d, J=5.3Hz, 3H).

Example 54: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyrazin-3(2H)-one (54)

Step 1: Preparation of 4-(trifluoromethyl)-5-(2S)-1-(3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-(2-(trimethylsilyl)ethoxy)methyl)pyrazin-3(2H)-one (54-a)

(S)-1-(2-((6-oxo-5-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-4-yl)amino)propyl)-1H-pyrrole-3-carboxylic acid (200 mg, 0.435 mmol) was dissolved in dichloromethane (10 mL), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (248 mg, 0.653 mmol), N,N-diisopropylethylamine (197 ul, 1.13 mmol) and 3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane hydrochloride (128 mg, 0.435 mmol) were added and reacted at room temperature overnight. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product 4-(trifluoromethyl)-5-(2S)-1-(3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-(2-(trimethylsilyl)ethoxy)methyl)pyrazin-3(2H)-one (54-a) (75 mg, yield 24.7%). ESI [M+H] ⁺ = 689.4

Step 2: Preparation of 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyrazin-3(2H)-one (54)

4-(Trifluoromethyl)-5-(2S)-1-(3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-(2-(trimethylsilyl)ethoxy)methyl)pyrazin-3(2H)-one (54-a) (75 mg, 0.107 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (6 mL) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane:methanol=15:1) to give the target product 4-(trifluoromethyl)-5-((2S)-1-(3-(5-(trifluoromethyl)pyrazin-2-yl)-3,8-diazabicyclo[3.2.1]octane-8-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyrazin-3(2H)-one (54) (40.2 mg, yield 65.8%). ESI[M+H] ⁺ =571.4, ESI[2M+H] ⁺ =1141.5. ¹ H NMR (600MHz, CD ₃ OD) δ 8.42 (s, 1H), 8.25 (s, 1H), 7.35 (s, 1H), 7.20 (s, 1H), 6.80 (s, 1H), 6.38 (s, 1H), 4.38 ( s, 1H), 4.31-4.17 (m, 3H), 3.96 (dd, J=13.4, 10.5Hz, 1H), 3.25 (t, J=13.6Hz, 3H), 2.05-1.87 (m, 3H), 1.79 (d, J=7.9Hz, 2H), 1.37 (d, J=6.1Hz, 3H).

Example 55: Preparation of (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (55)

Step 1: Preparation of (S)-1-(2-(tert-butyloxycarbonyl)(methyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (55-a)

(S)-1-(2-(tert-Butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (50 mg, 0.177 mmol) was dissolved in N,N-dimethylformamide (4 mL), sodium hydride (10 mg, 0.23 mmol) and iodomethane (17 ul, 0.265 mmol) were added at 0°C, and then reacted at 70°C overnight. Water was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the target product (S)-1-(2-(tert-Butyloxycarbonyl)(methyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (55-a) (50 mg, yield 96.2%). ESI[M-100+H]1=197.2, ESI[M-56+H] ⁺ =241.0, ESI[2M+H] ⁺ =593.4

Step 2: Preparation of (S)-1-(2-(tert-butyloxycarbonyl)(methyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (55-b)

(S)-1-(2-(tert-Butyloxycarbonyl)(methyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (55-a) (50 mg, 0.169 mmol) was dissolved in methanol (3.5 mL), and water (0.5 mL) and sodium hydroxide (14 mg, 0.339 mmol) were added, and the mixture was heated at 80°C in a sealed tube to react overnight. The pH was adjusted to 7 with 1N hydrochloric acid, and the reaction solution was directly concentrated under reduced pressure to obtain the target product (S)-1-(2-(tert-Butyloxycarbonyl)(methyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (55-b) (60 mg, yield 126.0%).

Step 3: Preparation of tert-butyl (S)-methyl (1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)carbamate (55-c)

(S)-1-(2-(tert-Butyloxycarbonyl)(methyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (55-b) (47 mg, 0.167 mmol) was dissolved in dichloromethane (5 mL), and 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (95 mg, 0.25 mmol), N,N-diisopropylethylamine (75 ul, 0.434 mmol) and 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine hydrochloride (44 mg, 0.167 mmol) were added and reacted at room temperature for 2 hours. After concentration under reduced pressure, the product was purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product, tert-butyl (S)-methyl (1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (65 mg, yield 78.8%). ESI [M+H] ⁺ = 497.3, ESI [M-56+H] ⁺ = 441.2, ESI [2M+H] + = 993.5, ESI [2M+Na] ⁺ = 1015.5

Step 4: Preparation of (S)-(1-(2-(methylamino)propyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (55-d)

Tert-butyl (S)-methyl (1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (65 mg, 0.131 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (0.7 mL) was added, and the mixture was reacted at room temperature for 2 hours. The reaction solution was evaporated under reduced pressure to dryness to obtain the target product (S)-(1-(2-(methylamino)propyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (55-d) (94 mg, yield 140.3%).

Step 5: Preparation of (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (55-e)

(S)-(1-(2-(Methylamino)propyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (55-d) (94 mg, 0.184 mmol) was dissolved in ethanol (5 mL), and triethylamine (103 ul, 0.737 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazine-3(2H)-one (61 mg, 0.184 mmol) were added, and the reaction was carried out at 60°C for 8 hours. After concentration under reduced pressure, the residue was dissolved in dichloromethane, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (55-e) (65 mg, yield 51.3%). ESI [M+H] ⁺ = 689.3

Step 6: Preparation of (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (55)

(S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-(2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (55-e) (65 mg, 0.094 mmol) was dissolved in 4M hydrochloric acid ethyl acetate solution (5 mL) and reacted at room temperature for 2 hours. After concentration under reduced pressure, the reaction mixture was purified on a preparative plate (dichloromethane:methanol=15:1) to give the target product (S)-5-(methyl(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (55) (15.6 mg, yield 29.6%). ESI[M+H]+=559.4, ESI[2M+H]+=1117.7.1H NMR (400MHz, CD3OD) δ8.61 (s, 2H), 7.36 (s, 1H), 7.16 (s, 1H), 6.77-6.71 (m, 1H), 6.30-6.24 (m, 1H), 4.31 (ddd, J=1 2.1, 8.3, 4.7Hz, 1H), 4.26-4.16 (m, 1H), 4.05 (dd, J=14.3, 3.6Hz, 1H), 4.01-3.90 (m, 4H), 3.84-3.72 (m, 4H), 3.06 (s, 3H), 1.40 (d, J=6.4Hz, 3H).

Example 56: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (56)

Step 1: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (56-a)

1H-pyrrole-3-carboxylic acid methyl ester (245 mg, 1.96 mmol) was dissolved in N,N-dimethylformamide (10 ml), cesium carbonate (1.92 g, 5.89 mmol) was added, and the mixture was stirred at room temperature for 30 minutes, and then (S)-(1-iodopropane-2-yl)carbamic acid tert-butyl ester (840 mg, 2.95 mmol) was added, and the mixture was stirred at 80°C overnight. Water (10 ml) was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a Flash column (dichloromethane: methanol = 25: 1) to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (56-a) (415 mg, yield 75.0%). ESI[M-56+H]+=227.1, ESI[2M+H]+=565.6, ESI[2M+Na]+=587.4

Step 2: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (56-b)

(S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid methyl ester (56-a) (200 mg, 0.709 mmol) was dissolved in methanol (10 ml), and water (3 ml) and sodium hydroxide (57 mg, 1.43 mmol) were added, and the mixture was stirred at 80°C in a sealed tube overnight. The pH of the reaction solution was adjusted to 7 with 1N hydrochloric acid, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (56-b) (180 mg, yield 94.7%). ESI[M-56+H]+=213.0, ESI[2M+H]+=537.4, ESI[2M+Na]+=559.4

Step 3: Preparation of tert-butyl (S)-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (56-c)

(S)-1-(2-((tert-Butyloxycarbonyl)amino)propyl)-1H-pyrrole-3-carboxylic acid (56-b) (18 mg, 0.067 mmol) was dissolved in dichloromethane (3 ml), 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylhexafluorouronium (38 mg, 0.10 mmol) was added, and stirred at room temperature for 15 minutes. Then 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (18 mg, 0.067 mmol) and N,N-diisopropylethylamine (29 ul, 0.168 mmol) were added and stirred at room temperature for 3 hours. Saturated ammonium chloride solution (3 ml) was added to quench the reaction, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product, tert-butyl (S)-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (56-c) (30 mg, yield 93.8%). ESI [M+H] + = 483.3, ESI [2M+H] + = 965.6, ESI [M-56+H] + = 427.3

Step 4: Preparation of (S)-(1-(2-aminopropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone trifluoroacetate (56-d)

Tert-butyl (S)-(1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (56-c) (30 mg, 0.062 mmol) was dissolved in dichloromethane (3 ml), trifluoroacetic acid (0.5 ml) was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the target product (S)-(1-(2-aminopropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone trifluoroacetate (56-d) (35 mg, yield 113.3%).

Step 5: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (56-e)

(S)-(1-(2-Aminopropyl)-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone trifluoroacetate (56-d) (30 mg, 0.060 mmol) was dissolved in ethanol (3 ml), and triethylamine (21 ul, 0.15 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (20 mg, 0.060 mmol) were added, and stirred at 60°C for 4.5 hours. The reaction solution was concentrated, and the residue was dissolved in dichloromethane, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (56-e) (28 mg, yield 68.6%). ESI[M] = 675.4, ESI[M-28] ⁺ = 647.3

Step 6: Preparation of (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (56)

(S)-4-(Trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (56-e) (28 mg, 0.042 mmol) was dissolved in hydrogen chloride ethyl acetate solution (3 ml, 4 mol/L) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and purified on a preparative plate (dichloromethane:methanol=20:1) to give the target product (S)-4-(trifluoromethyl)-5-((1-(3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)pyridazin-3(2H)-one (56) (13 mg, yield 57.5%). ESI[M+H] ⁺ =545.40, ¹ H NMR (400MHz, CDCl ₃ ) δ10.51 (s, 1H), 8.51 (s, 2H), 7.11 (d, J = 5.2Hz, 2H), 6.57 (t, J = 2.3Hz, 1H), 6.30 (s, 1H), 5.33-5.24 (m, 1H), 4 .11 (dd, J=19.9, 6.3Hz, 2H), 3.93 (s, 4H), 3.78 (d, J=5.2Hz, 4H), 3.64 (s, 1H), 1.42-1.37 (m, 3H).

Example 57: Preparation of (S)-5-((1-(3-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (57)

Step 1: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-4-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (57-a)

4-Methyl-1H-pyrrole-3-carboxylic acid ethyl ester (166 mg, 1.08 mmol) was dissolved in N,N-dimethylformamide (10 ml), cesium carbonate (1.06 g, 3.24 mmol) was added, and the mixture was stirred at room temperature for 30 minutes, and then (S)-(1-iodopropane-2-yl)carbamic acid tert-butyl ester (463 mg, 1.62 mmol) was added, and the mixture was stirred at 80°C overnight. Water (10 ml) was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a Flash column (dichloromethane: methanol = 20: 1) to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-4-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (57-a) (280 mg, yield 83.3%). ESI[M-56+H]+=255.1, ESI[M+H]+=311.2, ESI[2M+Na]+=643.5

Step 2: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-4-methyl-1H-pyrrole-3-carboxylic acid (57-b)

(S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-4-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (57-a) (140 mg, 0.451 mmol) was dissolved in ethanol (7 ml), and water (2 ml) and sodium hydroxide (36 mg, 0.902 mmol) were added, and the mixture was stirred at 80°C in a sealed tube overnight. The pH of the reaction solution was adjusted to 7 with 1N hydrochloric acid, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified with a Flash column (dichloromethane: methanol = 20: 1) to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-4-methyl-1H-pyrrole-3-carboxylic acid (57-b) (60 mg, yield 47.2%). ESI[M-56+H] ⁺ =227.1, ESI[2M+H] ⁺ =565.4

Step 3: Preparation of tert-butyl (S)-(1-(3-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (57-c)

(S)-1-(2-((tert-Butyloxycarbonyl)amino)propyl)-4-methyl-1H-pyrrole-3-carboxylic acid (57-b) (60 mg, 0.213 mmol) was dissolved in dichloromethane (5 ml), 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylhexafluorouronium (121 mg, 0.32 mmol) was added, and stirred at room temperature for 30 minutes. Then 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (74 mg, 0.213 mmol) and N,N-diisopropylethylamine (93 ul, 0.532 mmol) were added and stirred at room temperature for 3 hours. Saturated ammonium chloride solution (5 ml) was added to quench the reaction, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product, tert-butyl (S)-(1-(3-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (57-c) (70 mg, yield 66.4%). ESI [M+H] + = 497.3, ESI [2M+H] + = 993.6, ESI [M-56+H] + = 441.3

Step 4: Preparation of (S)-(1-(2-aminopropyl)-4-methyl-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone trifluoroacetate (57-d)

Tert-butyl (S)-(1-(3-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (57-c) (70 mg, 0.14 mmol) was dissolved in dichloromethane (5 ml), trifluoroacetic acid (0.5 ml) was added, and the mixture was stirred at room temperature for 1.5 hours. The reaction solution was concentrated under reduced pressure to obtain the target product (S)-(1-(2-aminopropyl)-4-methyl-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone trifluoroacetate (57-d) (75 mg, yield 104.2%).

Step 5: Preparation of (S)-5-((1-(3-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (57-e)

(S)-(1-(2-Aminopropyl)-4-methyl-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone trifluoroacetate (57-d) (75 mg, 0.15 mmol) was dissolved in ethanol (5 ml), and triethylamine (51 ul, 0.38 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (48 mg, 0.15 mmol) were added, and stirred at 60°C for 8 hours. The reaction solution was concentrated, and the residue was dissolved in dichloromethane, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 20: 1) to obtain the target product (S)-5-((1-(3-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (57-e) (55 mg, yield 54.3%). ESI [M+H] + = 689.4

Step 6: Preparation of (S)-5-((1-(3-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (57)

(S)-5-((1-(3-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (57-e) (55 mg, 0.080 mmol) was dissolved in hydrogen chloride ethyl acetate solution (5 ml, 4 mol/L) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and purified on a preparative plate (dichloromethane:methanol=18:1) to give the target product (S)-5-((1-(3-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (57) (20 mg, yield 44.8%). ESI[M+H]+=559.40, 1H NMR (400MHz, CDCl3) δ10.70 (s, 1H), 8.50 (s, 2H), 7.19 (s, 1H), 6.77 (s, 1H), 6.38 (s, 1H), 5.36-5.24 (m, 1H), 4.03 (d, J=11.6Hz, 2H) , 3.91 (s, 4H), 3.80 (dd, J=14.8, 8.0Hz, 1H), 3.67 (s, 4H), 2.06 (s, 3H), 1.37 (d, J=6.1Hz, 3H).

Example 58: Preparation of (S)-5-((1-(2-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (58)

Step 1: Preparation of 5-methyl-iH-pyrrole-3-carboxylic acid ethyl ester (58-a)

Dissolve 5-methyl-1H-pyrrole-3-carboxylic acid (100 mg, 1 M) in ethanol (5 mL), and add concentrated sulfuric acid (1.5 mL). React at 70°C for 5 hours. TLC (petroleum ether: ethyl acetate = 10: 1) shows that the raw material reacts completely and new spots are generated. Adjust the pH to neutral under ice bath, dry with anhydrous sodium sulfate, filter and concentrate under reduced pressure to obtain crude 5-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (58-a) (60 mg, yield 49.01%). ESI [M+H] ⁺ = 154.10

Step 2: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-5-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (58-b)

5-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (58-a) (60mg, 1eq) is added to N, N-dimethylformamide (6mL), and cesium carbonate (383mg, 3eq) and (S)-(1-iodopropane-2-yl) tert-butyl carbamate (167mg, 1.5eq) are added, and stirred for 16 hours at 80 ℃. The reaction is completed by LC-MS detection. The aqueous solution (40mL) is added to the reaction mixture. The mixed solution is extracted with ethyl acetate (15ml×3), and the combined organic layer is washed with salt water (20mlx2), dried over anhydrous sodium sulfate and filtered. The organic layer is concentrated under reduced pressure to provide a crude product. The crude product was purified by preparative plate (petroleum ether:ethyl acetate=4:1) to give the product (S)-ethyl 1-(2-((tert-butoxycarbonyl)amino)propyl)-5-methyl-1H-pyrrole-3-carboxylate (58-b) (33 mg, 27.14% yield), ESI [M+H] ⁺ =311.20.

Step 3: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-5-methyl-1H-pyrrole-3-carboxylic acid (58-c)

At 25°C, (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-5-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (58-b) (33 mg, 1 M), sodium hydroxide (8 mg, 2 M) were added to a solution of methanol (0.5 mL) and water (0.5 mL) and stirred for 16 hours. LC-MS monitored the reaction to be complete. The resulting mixture was concentrated under vacuum to give the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-5-methyl-1H-pyrrole-3-carboxylic acid (58-c) (35 mg, 80% purity, 93.28% yield). ESI[2M+H]+＝565.50

Step 4: Preparation of tert-butyl (S)-(1-(2-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (58-d)

(S)-1-(2-((tert-Butyloxycarbonyl)amino)propyl)-5-methyl-1H-pyrrole-3-carboxylic acid (58-c) (35 mg, 1 eq) was added to N,N-dimethylformamide (2 mL), and 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylhexafluorouronium (47 mg, 1 eq), 2-(piperazin-1-yl)-5-(trifluoromethyl)pyrimidine (35 mg, 1.2 eq) and N,N-diisopropylethylamine (48 mg, 3 eq) were added to the solution, and stirred at 25°C for 2 hours. The reaction was completed by LC-MS. Aqueous solution (50 mL) was added to the reaction mixture, and the mixed solution was extracted with ethyl acetate (10 ml×3), and the combined organic layer was washed with brine (10 ml×2), dried over anhydrous sodium sulfate and filtered. The organic layer was concentrated under reduced pressure to give solid tert-butyl (S)-(1-(2-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (58-d) (21 mg, 34.12% yield). ESI [M+H] + = 497.40

Step 5: Preparation of (S)-(1-(2-aminopropyl)-5-methyl-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (58-e)

Tert-butyl (S)-(1-(2-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (58-d) (21 mg, 1 M) was dissolved in dichloromethane (1 mL) at 0°C, followed by dropwise addition of trifluoroacetic acid (289 mg, 60 M). The reaction mixture was stirred at 25°C for 1 hour. The reaction was complete as determined by LC-MS. The resulting mixture was concentrated under vacuum to give a solid (S)-(1-(2-aminopropyl)-5-methyl-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (58-e) (30 mg, 50% purity, 89.47% yield). ESI [M+H] ⁺ = 397.30

Step 6: Preparation of (S)-5-((1-(2-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (58-f)

(S)-(1-(2-aminopropyl)-5-methyl-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (58-e) (30 mg, 50% purity, 1.00 M) was added to ethanol (1 mL), followed by the dropwise addition of triethylamine (11 mg, 3.00 M) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (15 mg, 1.2 M). The reaction mixture was stirred at 60 °C for 1 hour. LC-MS analysis showed that the starting material was completely consumed. The solid was filtered and the resulting mixture was concentrated under reduced pressure. The residue was purified by flash silica gel column chromatography (10 g) and polar ethyl acetate/petroleum ether (1:10) to give the product (S)-5-((1-(2-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (58-f) (25 mg, 95.93% yield). ESI [M+H]+=689.40

Step 7: Preparation of (S)-5-((1-(2-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (58)

(S)-5-((1-(2-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (58-f) (25 mg, 1 M) was added to 4N ethyl hydrochloride (3 mL). The reaction mixture was stirred at 25 ° C for 16 hours. The reaction was completed by LC-MS. The resulting mixture was concentrated under vacuum to give a crude product. The crude product was purified by preparative plate (dichloromethane: methanol = 10: 1) to give the product (S)-5-((1-(2-methyl-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (58) (6.6 mg, 32.56% yield). ESI [M+H] + = 559.30

Example 59: Preparation of (S)-5-((1-(2-methyl-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridin-3(2H)-one (59)

Step 1: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-2-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (59-a)

2-Methyl-1H-pyrrole-3-carboxylic acid ethyl ester (200 mg, 1.31 mmol) was dissolved in N,N-dimethylformamide (10 ml), cesium carbonate (1.28 g, 3.93 mmol) was added, and the mixture was stirred at room temperature for 30 minutes, and then (S)-(1-iodopropane-2-yl)carbamic acid tert-butyl ester (485 mg, 1.70 mmol) was added, and the mixture was stirred at 80°C overnight. Water (10 ml) was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a Flash column (dichloromethane: methanol = 20: 1) to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-2-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (59-a) (250 mg, yield 61.7%). ESI[M-56+H] ⁺ =255.1, ESI[M+H] ⁺ =311.3

Step 2: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-2-methyl-1H-pyrrole-3-carboxylic acid (59-b)

(S)-1-(2-((tert-Butyloxycarbonyl)amino)propyl)-2-methyl-1H-pyrrole-3-carboxylic acid ethyl ester (59-a) (250 mg, 0.806 mmol) was dissolved in ethanol (10 ml), and water (2 ml) and sodium hydroxide (65 mg, 1.63 mmol) were added, and the mixture was stirred at 80°C in a sealed tube overnight. The pH of the reaction solution was adjusted to 7 with 1N hydrochloric acid, and the remaining raw materials were washed out with dichloromethane. The aqueous phase was directly concentrated under reduced pressure to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-2-methyl-1H-pyrrole-3-carboxylic acid crude product (150 mg). The dichloromethane layer was concentrated under reduced pressure and then prepared into a plate (dichloromethane: methanol = 20: 1) to obtain the target product (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-2-methyl-1H-pyrrole-3-carboxylic acid (59-b) (26 mg, total yield 77.5%). ESI [M-100+H] + = 183.2, ESI [M-56+H] + = 227.1, ESI [2M+H] + = 565.4

Step 3: Preparation of tert-butyl (S)-(1-(2-methyl-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (59-c)

(S)-1-(2-((tert-Butyloxycarbonyl)amino)propyl)-2-methyl-1H-pyrrole-3-carboxylic acid (59-b) (26 mg, 0.092 mmol) was dissolved in dichloromethane (4 ml), 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylhexafluorouronium (53 mg, 0.14 mmol) was added, and stirred at room temperature for 30 minutes. Then 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (25 mg, 0.092 mmol) and N,N-diisopropylethylamine (40 ul, 0.23 mmol) were added, and the mixture was stirred at room temperature overnight. Saturated ammonium chloride solution (5 ml) was added to quench the reaction, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 25: 1) to obtain the target product, tert-butyl (S)-(1-(2-methyl-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (59-c) (21 mg, yield 46.0%). ESI [M+H] ⁺ = 497.4, ESI [2M+H] ⁺ = 993.7, ESI [M-56+H] ⁺ = 441.3

Step 4: Preparation of (S)-(1-(2-aminopropyl)-2-methyl-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone trifluoroacetate (59-d)

Tert-butyl (S)-(1-(2-methyl-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (59-c) (37 mg, 0.075 mmol) was dissolved in dichloromethane (4 ml), trifluoroacetic acid (0.5 ml) was added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the target product (S)-(1-(2-aminopropyl)-2-methyl-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone trifluoroacetate (59-d) (50 mg, yield 131.6%).

Step 5: Preparation of (S)-5-((1-(2-methyl-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (59-e)

(S)-(1-(2-Aminopropyl)-2-methyl-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone trifluoroacetate (59-d) (50 mg, 0.098 mmol) was dissolved in ethanol (5 ml), and triethylamine (34 ul, 0.25 mmol) and 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (32 mg, 0.098 mmol) were added, and stirred at 60°C for 8 hours. The reaction solution was concentrated, and the residue was dissolved in dichloromethane, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified on a preparative plate (dichloromethane: methanol = 20: 1) to give the target product (S)-5-((1-(2-methyl-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (59-e) (27 mg, yield 40.0%).

Step 6: Preparation of (S)-5-((1-(2-methyl-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (59)

(S)-5-((1-(2-methyl-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (59-e) (27 mg, 0.039 mmol) was dissolved in hydrogen chloride ethyl acetate solution (3 ml, 4 mol/L) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and then purified on a preparative plate (dichloromethane: methanol = 15: 1) to obtain the target product (S)-5-((1-(2-methyl-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (59) (11 mg, yield 50.2%). ESI [M+H]+ = 559.20

Example 60: Preparation of (S)-5-((1-(2-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (60)

Step 1: Preparation of methyl 2-fluoro-1H-pyrrole-3-carboxylate (60-a)

At room temperature, 1H-pyrrole-3-carboxylic acid methyl ester (3.5g, 27.97mmol), 1-chloromethyl-4-fluoro-1,4-diazobicyclo[2.2.2]octane bis(tetrafluoroborate) (11.88g, 33,56mmol) and potassium phosphate (8.9g, 41.95mmol) were dissolved in tetrahydrofuran (100mL). After the addition was completed, the reaction solution was stirred at 50°C overnight. The reaction solution was concentrated under reduced pressure and diluted with ethyl acetate (20mL), and the organic phase was washed with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain 2-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (60-a, 638mg, yield 15.9%). ESI[M+H] ⁺ = 311.1

Step 2: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-2-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (60-b)

At room temperature, methyl 2-fluoro-1H-pyrrole-3-carboxylate (60-a, 500 mg, 3.49 mmol) was dissolved in N,N-dimethylformamide (15 mL). Cesium carbonate (3.41 g, 10.47 mmol) and (S)-2-((tert-butyloxycarbonyl)amino)propyl 4-toluenesulfonate (3.41 g, 10.47 mmol) were added under argon replacement and stirring. After the addition was completed, the reaction solution was stirred at 70°C for 16 hours. The reaction solution was quenched by adding water (10 mL) and extracted with ethyl acetate (3*10 mL). The organic phases were combined and concentrated under reduced pressure. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 5: 1-1: 1) to give ((S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-2-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (60-b, 630 mg, yield: 60.1%). ESI [M+H] ⁺ = 311.0

Step 3: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-2-fluoro-1H-pyrrole-3-carboxylic acid (60-c)

At room temperature, ((S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-2-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (60-b, 653 mg, 2.28 mmol) was dissolved in methanol (10 mL) and water (3 mL), and potassium hydroxide (383 mg, 6.84 mmol) was added under stirring. After the addition was completed, the reaction solution was heated at 60°C and stirred for 12 hours. The reaction solution was adjusted to pH = 6-7 with hydrochloric acid and extracted with ethyl acetate (10 mL x 3). The organic phase was washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-2-fluoro-1H-pyrrole-3-carboxylic acid (60-c, 603 mg). ESI [M+H] + = 297.1

Step 4: Preparation of tert-butyl-(1-(2-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (60-d)

At room temperature, (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-2-fluoro-1H-pyrrole-3-carboxylic acid (60-c, 603 mg, 2.11 mmol), 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (489.5 mg, 2.11 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (801 mg, 2.11 mmol) and diisopropylethylamine (823 mg, 6.33 mmol) were dissolved in N,N-dimethylformamide (10 mL). After the addition was completed, the reaction solution was stirred at room temperature for 1 hour. The reaction solution was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phase was washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (dichloromethane: methanol = 10: 1) to obtain tert-butyl-(1-(2-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (60-d, 503 mg, yield: 47.7%). ESI [M+H] + = 510.1

Step 5: Preparation of (S)-(1-(2-aminopropyl)-5-fluoro-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (60-e)

At room temperature, tert-butyl-(1-(2-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)carbamate (60-d, 203 mg, 0.406 mmol) was dissolved in dichloromethane (20 mL), and 4M dioxane hydrochloride (10 mL) was added under stirring. After the addition was completed, the reaction solution was stirred at room temperature for 1 hour, and the reaction solution was concentrated under reduced pressure to obtain (S)-(1-(2-aminopropyl)-5-fluoro-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (60-e, 180 mg, crude product). ESI[M+H]+＝410.2

Step 6: Preparation of (S)-5-((1-(2-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (60-f)

At room temperature, (S)-(1-(2-aminopropyl)-5-fluoro-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (60-e, 180 mg, 0.45 mmol), 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (444.2 mg, 1.35 mmol) and triethylamine (136.6 mg, 1.35 mmol) were dissolved in ethanol (10 mL). After the addition was completed, the reaction solution was stirred at 60° C. for 1 hour, and the liquid phase mass spectrometry showed that the reaction was complete. The reaction solution was concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate = 0: 1) to give ((S)-5-((1-(2-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (60-f, 82 mg, yield: 26.3%). ESI [M+H] + = 702.3

Step 7: Preparation of (S)-5-((1-(2-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (60)

At room temperature, ((S)-5-((1-(2-fluoro-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (60-f, 82 mg, 0.118 mmol) was dissolved in dichloromethane (3 mL). Trifluoroacetic acid (2 mL) was added under stirring. After the addition was complete, After completion, the reaction solution was stirred at room temperature for 16 hours, and the liquid phase mass spectrometry showed that the reaction was complete. The reaction solution was concentrated under reduced pressure, and the crude product was prepared by reverse phase to obtain (S)-5-((1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (60) (3.13 mg, yield 4.69%). ESI [M+H] + = 563.3, 1H NMR (400MHz, CDCl3) δ10.87 (s, 1H), 8.53 (s, 2H), 7.28 (s, 1H), 6.72 (s, 1H), 5.70 (s, 1H), 5.35 (s, 1H), 3.87 (d, J=75.6Hz, 11H), 1.36 (d, J=41.2Hz, 3H).

Example 61: Preparation of (S)-5-((1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (61)

Step 1: Preparation of 5-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (61-a)

At room temperature, 1H-pyrrole-3-carboxylic acid methyl ester (3.5 g, 27.97 mmol), 1-chloromethyl-4-fluoro-1,4-diazobicyclo[2.2.2]octane bis(tetrafluoroborate) (11.88 g, 33,56 mmol) and potassium phosphate (8.9 g, 41.95 mmol) were dissolved in tetrahydrofuran (100 mL). After the addition was completed, the reaction solution was stirred at 50°C overnight. The reaction solution was concentrated under reduced pressure and diluted with ethyl acetate (20 mL), and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain 5-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (61-a, 638 mg, yield 15.9%). ESI [M+H] + = 311.1

Step 2: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-5-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (61-b)

At room temperature, 5-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (61-a, 500 mg, 3.49 mmol) was dissolved in N,N-dimethylformamide (15 mL). Cesium carbonate (3.41 g, 10.47 mmol) and (S)-2-((tert-butyloxycarbonyl)amino)propyl 4-toluenesulfonate (3.41 g, 10.47 mmol) were added under argon replacement and stirring. After the addition was completed, the reaction solution was stirred at 70°C for 16 hours. The reaction solution was quenched by adding water (10 mL) and extracted with ethyl acetate (3*10 mL). The organic phases were combined and concentrated under reduced pressure. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 5: 1-1: 1) to give ((S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-5-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (61-b) (630 mg, yield 60.1%). ESI [M+H] + = 311.0

Step 3: Preparation of (S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-5-fluoro-1H-pyrrole-3-carboxylic acid (61-c)

At room temperature, ((S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-5-fluoro-1H-pyrrole-3-carboxylic acid methyl ester (61-b, 653 mg, 2.28 mmol) was dissolved in methanol (10 mL) and water (3 mL), and potassium hydroxide (383 mg, 6.84 mmol) was added under stirring. After the addition was completed, the reaction solution was heated at 60°C and stirred for 12 hours. The reaction solution was adjusted to pH = 6-7 with hydrochloric acid and extracted with ethyl acetate (10 mL x 3). The organic phase was washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give (S)-1-(2-((tert-butoxycarbonyl)amino)propyl)-5-fluoro-1H-pyrrole-3-carboxylic acid (61-c, 603 mg). ESI [M+H] ⁺ = 297.1

Step 4: Preparation of tert-butyl-(1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (61-d)

At room temperature, ((S)-1-(2-((tert-butyloxycarbonyl)amino)propyl)-5-fluoro-1H-pyrrole-3-carboxylic acid (61-c, 603 mg, 2.11 mmol), 2-(piperazine-1-yl)-5-(trifluoromethyl)pyrimidine (489.5 mg, 2.11 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (801 mg, 2.11 mmol) and diisopropylethylamine (823 mg, 6.33 mmol) were dissolved in N,N-dimethylformamide (10 mL). After the addition was completed, the reaction solution was stirred at room temperature for 1 hour. The reaction solution was diluted with water (10 mL) and ethyl acetate (10 mL) was added. x3) extraction. The organic phase was washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (dichloromethane: methanol = 10: 1) to obtain tert-butyl-(1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)carbamate (61-d, 503 mg, yield: 47.7%). ESI [M+H] ⁺ = 510.1

Step 5: Preparation of (S)-(1-(2-aminopropyl)-5-fluoro-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (61-e)

At room temperature, tert-butyl-(1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propane-2-yl)carbamate (61-d, 203 mg, 0.406 mmol) was dissolved in dichloromethane (10 mL), and 4M dioxane hydrochloride (10 mL) was added under stirring. After the addition was completed, the reaction solution was stirred at room temperature for 1 hour, and the reaction solution was concentrated under reduced pressure to obtain (S)-(1-(2-aminopropyl)-5-fluoro-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-yl)methanone (61-e, 180 mg, crude product). ESI[M+H] ⁺ =410.2

Step 7: Preparation of (S)-5-((1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (61-f)

At room temperature, (S)-(1-(2-aminopropyl)-5-fluoro-1H-pyrrol-3-yl)(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)methanone (61-e, 180 mg, 0.45 mmol), 5-chloro-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (444.2 mg, 1.35 mmol) and triethylamine (136.6 mg, 1.35 mmol) were dissolved in ethanol (10 mL). After the addition was completed, the reaction solution was stirred at 60°C for 1 hour, and the liquid phase mass spectrometry showed that the reaction was complete. The reaction solution was concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate = 0: 1) to obtain (S)-5-((1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (61-f, 82 mg, yield 26.3%). ESI [M+H] + = 702.3

Step 8: Preparation of (S)-5-((1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (61)

At room temperature, ((S)-5-((1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (61-f, 82 mg, 0.118 mmol) was dissolved in dichloromethane (3 mL). Trifluoroacetic acid (2 mL) was added under stirring. After the addition was completed, Afterwards, the reaction solution was stirred at room temperature for 16 hours, and the liquid phase mass spectrometry showed that the reaction was complete. The reaction solution was concentrated under reduced pressure, and the crude product was prepared by reverse phase to obtain (S)-5-((1-(2-fluoro-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)-1H-pyrrol-1-yl)propan-2-yl)amino)-4-(trifluoromethyl)pyridazin-3(2H)-one (61) (10.87 mg, yield 16.3%). ESI [M+H] + = 563.1.1H NMR (400MHz, CDCl3) δ 10.73 (s, 1H), 8.59 (s, 2H), 6.70 (s, 1H), 5.68 (s, 1H), 5.33 (s, 1H), 4.20-3.66 (m, 11H), 1.42 (s, 3H).

Effect Example 1: PARP7 Inhibitory Activity Assay

The in vitro activity of the above compounds demonstrated in the following test that PARP7 can catalyze NAD-dependent ADP ribosylation. The PARP7 enzyme activity test method used in the present disclosure adopts the PARP7 chemiluminescent activity assay kit reagent of BPS Bioscience, USA. The test was carried out in a 384-well plate and was divided into three steps, which are briefly described as follows.

first step:

1) Dilute the 5x histone mixture 1:5 with PBS, add 20 μl to each well, and incubate at 4°C overnight;

2) Wash each well three times with 100 μl PBST (1x PBS, 0.05% Tween-20) and pat dry on paper;

3) Add 80 μl of blocking buffer 3 to each well and incubate at room temperature for 90 minutes;

4) Wash each well three times with 100 μl PBST and pat dry on paper.

Step 2:

1) Dilute 10x PARP assay buffer 10-fold with water;

2) Thaw PARP7 enzyme on ice, add 8 μl PARP7 enzyme to each well of the positive control group and the test compound group, and add 8 μl 1x PARP7 buffer to each well of the blank control group;

3) Add 10 μl of substrate to each well;

4) Add 2 μl of test compound or positive control;

5) Blank plus 8 μl 1X PARP7 buffer;

6) Add 8 μl of diluted PARP7 to the positive control and test wells and incubate at room temperature for 1 hour;

7) After 1 hour, the reaction mixture was discarded, washed three times with 100 μl PBST buffer, and patted dry.

third step:

1) Dilute horseradish peroxidase-labeled streptavidin (Streptavidin-HRP) at 1:50 with blocking buffer 3, add 20 μl to each well, and incubate at room temperature for 30 minutes;

2) Wash three times with 100 μl PBST buffer and pat dry;

3) Mix ELISA ECL Substrate A + ELISA ECL Substrate B on ice, 40 μl per well;

4) The enzyme activity was tested by reading the plate on a multifunctional microplate reader Spark 10M (Tecan Company) and the IC ₅₀ value was calculated. The test results are shown in Table 2.

Biological Activity

For _IC50 values, "++++" indicates _IC50 <50 nM; "+++" indicates _IC50 between 50 nM and 200 nM; "++" indicates _IC50 between 200 and 1000 nM; and "+" indicates _IC50 >1000 nM.

Table 2 In vitro biological activity IC50 values (nM) of the compounds in each example

Claims (22)

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, in, U ₁ is N; U ₂ is CH; U ₄ is N; Selected from , , , , , , , , , , , , , or ; R ₁ is selected from hydrogen, deuterium or C _1-3 alkyl, wherein the C _1-3 alkyl is optionally substituted with one or more halogens; U ₃ is selected from -NH-, -N(R ₂ )-, O or S; Wherein, R ₂ is selected from C _1-3 alkyl, C _3-6 cycloalkyl or C _3-6 heterocycloalkyl; R ₃ and R ₄ are each independently selected from hydrogen, deuterium, C _1-3 alkyl, or R ₃ and R ₄ are connected to each other to form a divalent C _3-6 cycloalkyl or a 3-7 membered heterocycloalkyl containing at least one heteroatom selected from N, O or S, wherein the C _1-3 alkyl, C _3-6 cycloalkyl or 3-7 membered heterocycloalkyl is optionally substituted with one or more halogen, hydroxy, amino, nitro, cyano; R ₅ is selected from hydrogen, halogen or C _1-3 alkyl; R ₆ and R _6a are each independently selected from hydrogen, chlorine or methyl, or R ₆ and R _6a are connected to form phenyl or pyridyl, and the phenyl or pyridyl is optionally substituted by one or more fluorine or chlorine; R ₈ , R _8a , R ₉ , R _9a , R ₁₀ , R _10a , R ₁₁ , R _11a are each independently selected from hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano or C _1-6 alkyl, wherein the C _1-6 alkyl is optionally substituted with one or more halogen, hydroxyl, cyano, carboxyl, sulfonamido, C _1-3 alkyl, C _1-3 amide, -C(O)-O-CH ₃ or -C(O)-O-CH ₂ CH ₃ , or any two of R ₈ , R ₉ , R ₁₀ , R ₁₁ are connected to each other to form -(CH ₂ ) _n2 -, wherein n ₂ is 1, 2 or 3; Wherein, the compound of formula (I) is not . 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₁ is selected from hydrogen, deuterium or methyl, and the methyl is optionally substituted by one or more halogens. 3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₁ is selected from hydrogen, deuterium, methyl or trifluoromethyl. 4. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein _R2 is selected from methyl, ethyl, isopropyl, cyclopropyl, and cyclohexyl. 5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein _R2 is selected from methyl. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein U ₃ is selected from -NH-, O, S, or -N(CH ₃ )-. 7. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R ₃ and R ₄ are each independently selected from hydrogen, deuterium, C _1-3 alkyl, or R ₃ and R ₄ are connected to each other to form a divalent C _5-6 cycloalkyl or a 5-7 membered heterocycloalkyl containing at least one heteroatom selected from N, O or S, and the C _1-3 alkyl C _3-6 cycloalkyl is optionally substituted by one to three halogens. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R ₃ and R ₄ are each independently selected from hydrogen, deuterium, methyl, trifluoromethyl, or R ₃ and R ₄ are linked to form a divalent cyclopentyl, cyclohexyl or tetrahydrofuranyl group. 9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein _R5 is selected from hydrogen or methyl. 10. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R ₅ is hydrogen. 11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Selected from , , , , , or . 12. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₈ , R _8a , R ₉ , R _9a , R ₁₀ , R _10a , R ₁₁ , and R _11a are each independently selected from hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, and neopentyl, and the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, and neopentyl are optionally substituted with one, two or three halogen, hydroxyl, cyano, carboxyl, formamido, sulfonamido, methyl, ethyl, propyl, isopropyl, -C(O)-O-CH ₃ or -C(O)-O-CH ₂ CH ₃ , or R ₈ , R ₉ , R ₁₀ , R 10a , R 11a are each independently selected from hydrogen, deuterium, halogen, hydroxyl, amino, nitro, cyano, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, and neopentyl. Any two of ₁₁ are linked to each other to form -(CH ₂ ) _n2 -, wherein _n2 is 1, 2 or 3. 13. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₈ , R _8a , R ₉ , R _9a , R ₁₀ , R _10a , R ₁₁ , R _11a are each independently selected from hydrogen, deuterium, halogen, methyl or ethyl, the methyl or ethyl being optionally substituted by one or two cyano groups, formamide groups or -C(O)-O-CH ₂ CH ₃ , or any two of R ₈ , R ₉ , R ₁₀ , R ₁₁ are connected to each other to form -(CH ₂ ) _n2 -, wherein n ₂ is 1, 2 or 3. 14. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Selected from , , , , , , , , , , , , , . 15. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which has a compound represented by formula (II): Wherein, _n1 , _U1 , _U2 , _U3 , _U4 , U5, _U6 , _U7 , _U8 , _R3 , _R4 , _R6 , _R6a , _R8 , _R8a , _R9 , _R10 and _{R12 are} as defined in claim 1. 16. The compound or pharmaceutically acceptable salt thereof according to claim 15, which is a compound represented by formula (IIIa), (IIIc), (IIId) or (IIIe): in, n ₄ is selected from 0 or 1; _X5 is selected from _CH2 or O; The definitions of n ₁ , U ₅ , U ₆ , U ₈ , R ₃ , R ₄ , R ₈ , R _8a , R ₉ , R ₁₀ and R ₁₂ are as described in claim 15 . 17. The following compounds or pharmaceutically acceptable salts thereof, . 18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier. 19. Use of the compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 18, in the preparation of a medicament for treating a PARP7-mediated disorder in a patient. 20. The use according to claim 19, wherein the PARP7-mediated disease includes cancer, immune disease, inflammation and viral infection. 21. Use of the compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 18 in the preparation of a PARP7 inhibitor. 22. A method for inhibiting PARP7 in a biological sample for non-disease diagnosis or treatment purposes, comprising contacting the biological sample with the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 17 or the pharmaceutical composition according to claim 18.