Compounds having antitumor activity and their use

Abstract

The present invention provides a compound with antitumor activity and its use. As a PRMT5 inhibitor, the compound of the present invention has the structure shown in formula (I). Experiments have confirmed that the compound of the present invention has a strong inhibitory effect on PRMT5 activity and tumor cell proliferation, and is considered to be a promising compound for treating diseases mediated by PRMT5. In addition, the present invention also researches a specific synthesis method, which is simple in process, convenient in operation, and favorable for large-scale industrial production and application. [Formula 1] TIFF2024517495000421.tif50170

Description

Technical Field

[0001] Cross-reference to Related Applications This application claims priority to Chinese Patent Application No. 202110521548.0, filed in China on May 13, 2021, with the title "Compound Having Antitumor Activity and Use Thereof", the content of which is incorporated herein by reference in its entirety.

[0002] The present invention relates to the field of pharmaceutical technology, and specifically, to compounds as PRMT5 inhibitors, methods for preparing such compounds, and uses thereof.

Background Art

[0003] PRMT5 is a kind of protein arginine methyltransferase (PRMTs), an abbreviation of the English name Protein arginine N-methyltransferase 5, and is a new antitumor target related to epigenetic modification. It has several other names such as Hsl7, Jbp1, Skb1, Capsuleen, or Dart5. PRMT5 is the main enzyme for monomethylation and symmetric dimethylation of arginine. More and more literature has shown that protein arginine methyltransferases (PRMTs) play important roles in different biological processes such as cell growth and proliferation, apoptosis, and metastasis.

[0004] The role of protein arginine methyltransferase is to transfer one methyl group from S-adenosylmethionine (or AdoMet, or SAM) to an arginine residue on histone or other proteins, forming methylarginine and S-adenosylhomocysteine (SAH). Currently, nine members of this family (PRMT 1 - 9) have been identified, and PRMTs can be divided into three types according to the different ways of catalyzing arginine methylation. Type I PRMTs include PRMT1, PRMI2, PRMT3, PRMT4, PRMT6 and PRM8, which catalyze monomethylarginine (MMA) and asymmetric dimethylarginine (aDMA). Type II PRMTs include PRMT5 and PRMT9, which catalyze MMA and symmetric dimethylarginine (sDMA). Type III PRMTs is PRMT7, which can only catalyze MMA. As an epigenetic enzyme, PRMT5 can symmetrically methylate arginine residues of histone or non-histone substrates, affect multiple target genes and multiple signaling pathways, and play an important role in protein methylation. For example, it is involved in alternative splicing, post-transcriptional regulation, RNA processing, cell proliferation, cell differentiation, cell apoptosis, tumor formation, etc. Selective inhibition of PRMT5 can potentially be a powerful new anti-cancer drug. Research on the development of new drugs targeting PRMT5 plays a positive role in filling the gap in solving unmet clinical needs.

[0005] In the past few years, as can be seen from WO2014100719A, WO2019102494A, WO2015200677A, WO2015200680A, WO2014100764A, WO2014100730A, WO2014100716A, WO2014100695A, WO2019173804A, CN108570059A, WO2018167269A, etc., there have been more reports on PRMT5 inhibitors. And two compounds, JNJ-64619178 and GSK-3326595, have already been used in the clinical treatment of solid tumors and mantle cell lymphoma. [Chemical]

[0006] JNJ-64619178 is a selective PRMT5 inhibitor developed by Johnson & Johnson and has the function of inhibiting the growth of various tumor cells in vitro. Johnson & Johnson selected a large number of xenograft animal models to demonstrate the effectiveness of its anti-tumor function. For example, anti-tumor experiments were conducted by selecting xenograft models of small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), acute myeloid leukemia (AML), and non-Hodgkin lymphoma. In the models, significant tumor growth inhibition up to 99% was observed, and tumor growth continued to be suppressed within several weeks after drug withdrawal. JNJ-64619178 can inhibit the Sym-Arg dimethylation of SMD1 / 3 protein, a core component of tumor extracts, and can also inhibit the Sym-Arg dimethylation of serum proteins. These can be used as pharmacodynamic markers for PRMT5 to inhibit tumor growth in xenograft models. In the SCLC model, an effective and long-term inhibitory effect of PRMT5 on SMD1 / 3 dimethylation was observed during and after administration. Based on these high selectivities and effectiveness, good pharmacokinetics and safety, and obvious preclinical therapeutic effects and pharmacodynamic results, JNJ-64619178 initiated a Phase I clinical trial in 2018.

[0007] GSK-3326595 is an optimized version of EPZ015666 (the structure is as follows). It is a highly selective, orally available small molecule and the first-generation PRMT5 inhibitor. EPZ015666 shows significant in vivo and in vitro activity against mantle cell lymphoma. GlaxoSmithKline (GSK) announced in September 2016 that GSK-3326595 first entered clinical trials after two years of optimization and preclinical research. At the 2019 ESMO Congress, GlaxoSmithKline presented the clinical phase I data of GSK-3326595. In the phase I clinical trial of GSK-3326595, adult solid tumor patients were selected, and the main objectives were to conduct safety, tolerance, and PK / PD tests and collect efficacy data (ORR and DCR). The data showed that GSK 3326595 PK was dose-dependent in plasma.

Chemical Structure

[0008] Although several small molecule PRMT5 inhibitors have been disclosed, currently, no PRMT5 inhibitor is commercially available. Therefore, there is an urgent need to develop new compounds with the potential for market launch and better efficacy and pharmacokinetic results.

Summary of the Invention

Problems to be Solved by the Invention

[0009] The object of the present invention is to provide a compound having a completely new structure as a PRMT5 inhibitor, a method for preparing the compound, and its use in the treatment of diseases mediated by PRMT5.

Means for Solving the Problems

[0010] The first aspect of the present invention provides a compound represented by the following formula (I), and its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates, or isotope-labeled analogs.

Chemical Structure

[0011] In the formula, L 1 and L 2 are each independently selected from -C(R 1 )(R 2 )-, -C(R 1 )(R 2 )C(R 1 )(R 2 )-, -C(R 1 )(R 2 )C(R 1 )(R 2 )C(R 1 )(R 2 )-, provided that R 1 , R 2 are each independently selected from hydrogen, halogen, hydroxy group, mercapto group, amino group, cyano group, C 1-6 alkyl group, C 3-6 cycloalkyl group, 4- to 6-membered heterocyclic group, -OR 3 , -NHR 3 , -NR 3 R 4 in each occurrence, and R 3 , R 4 are each independently selected from C 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3-6 cycloalkyl group, 4- to 6-membered heterocyclic group, aryl group, 5- to 6-membered heteroaryl group, X is selected from C(R 5 ), or N, provided that R 5 is each independently selected from hydrogen, halogen, hydroxy group, mercapto group, amino group, cyano group in each occurrence, Y is -(chemical bond), -H, -OH, -NH2, halogen, -O-, -S-, -CO-, -C(R 6 )F-, -CF2-, -SO-, -SO2-, -(CH2) p N(R 6 ), -N(R 6 )(CH2) p -, -S(O)N(R 6 ), -S(O)2N(R 6)-, -N(R 6 )SO-, -N(R 6 )S(O)2-, -C(O)N(R 6 )-, -N(R 6 )C(O)-, -CH(R 6 ) is one selected from the group consisting of, provided that p = 0, 1, 2, or 3, and R 6 is hydrogen, an optionally substituted C 1-6 alkyl group, an optionally substituted C 3-6 cycloalkyl group, or an optionally substituted 4- to 6-membered heterocyclic group, and said optional substitution means that the hydrogen on the group to be substituted is unsubstituted, or one or more substitutable sites of the group to be substituted are independently substituted with a substituent selected from the group consisting of halogen, hydroxy group, mercapto group, amino group, cyano group, C 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3-6 cycloalkyl group, 4- to 6-membered heterocyclic group, aryl group, 5- to 6-membered heteroaryl group, and when Y is selected from -H, -OH, -NH2, halogen, G 4 does not exist, Z is —(chemical bond), -O-, -S-, -CO-, -N(R 7 )-, -S(O)N(R 7 )-, -S(O)2N(R 7 )-, -N(R 7 )SO-, -N(R 7 )S(O)2-, -C(O)N(R 7 )-, -N(R 7 )C(O)-, -N(R 7 )C(O)N(R 7 )-, -CH(R 7 )-, -CH(OH)-, -CH(CF3)-, -CH(NHR 7 )-, -C=N(R 7 )-, -SO-, -SO2-, -CF(R 7 )-, CF2, [Chemical formula] is one selected from the group consisting of, provided that R 7is, independently at each occurrence, hydrogen, optionally substituted C 1-6 alkyl group, optionally substituted C 3-6 cycloalkyl group, optionally substituted C 1-6 alkoxy group, or one selected from optionally substituted 4- to 6-membered heterocyclic groups, where the optional substitution means that the hydrogen on the group to be substituted is unsubstituted, or one or more substitutable sites on the group to be substituted are independently substituted with a substituent selected from halogen, hydroxy group, mercapto group, amino group, cyano group, C 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3-6 cycloalkyl group, 4- to 6-membered heterocyclic group, aryl group, 5- to 6-membered heteroaryl group, G 1 is, independently at each occurrence, hydrogen, halogen, hydroxy group, mercapto group, amino group, cyano group, C 1-5 linear alkyl group or branched alkyl group, G 2 is one selected from halogen, hydroxy group, mercapto group, amino group, cyano group, optionally substituted R 8 , optionally substituted -O(R 8 ), optionally substituted -S(R 8 ), optionally substituted -NH(R 8 ), optionally substituted -N(R 8 )(R 8 ), provided that R 8 is, independently at each occurrence, one selected from C 1-6 alkyl group, C 3-6 cycloalkyl group, 4- to 6-membered heterocyclic group, where the optional substitution means that the hydrogen on the group to be substituted is unsubstituted, or one or more substitutable sites on the group to be substituted are independently substituted with a substituent selected from halogen, hydroxy group, mercapto group, amino group, cyano group, C 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3-6 cycloalkyl group, 4- to 6-membered heterocyclic group, aryl group, 5- to 6-membered heteroaryl group, G 3is optionally substituted C 6-10 an aryl group, an optionally substituted 5- to 10-membered heteroaryl group, or an optionally substituted C 3-7 cycloalkyl group, or an optionally substituted 4- to 10-membered heterocyclic group, and the optional substitution means that the hydrogen on the group to be substituted is unsubstituted, or one or more substitutable sites of the group to be substituted are independently substituted with R 16 wherein R 16 is, independently at each occurrence, hydrogen, deuterium, halogen, hydroxy, mercapto, amino, cyano, -R 9 , -OR 9 , -OR 9 -N(R 9 )(R 10 ), -OR 9 -OR 10 , -SR 9 , SO(R 9 ), -SO2(R 9 ), -COOR 9 , -NH(R 9 ), -N(R 9 )(R 10 ), -N(R 9 )(R 10 )-N(R 9 )(R 10 ), -CONHR 9 , -CON(R 9 )(R 10 ), -SONH(R 9 ), -SON(R 9 )(R 10 ), SO2NH(R 9 ), -SO2N(R 9 )(R 10 ), and is selected from the group consisting of, provided that R 9 , R 10 is, independently at each occurrence, selected from hydrogen, or hydrogen, halogen, hydroxy, mercapto, amino, cyano, nitro, methoxy, -COOCH3, -COCH3, -COOC(CH3)3, -CH2-N(CH3)(CH3), -N(CH3)(CH3), -NH(CH3), C 1-6 alkyl group, C 2-6 alkenyl group, C 1-6 alkoxy group, C 3-6A C optionally substituted by one or more of a cycloalkyl group, a 4- to 6-membered heterocyclic group, an aryl group, and a 5- to 6-membered heteroaryl group 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3-10 selected from a cycloalkyl group, a 4- to 10-membered heterocyclic group, an aryl group, and a 5- to 10-membered heteroaryl group, G 4 is one selected from -SO2(CH3), an optionally substituted C 1-12 alkyl group, an optionally substituted C 2-12 alkenyl group, an optionally substituted C 2-12 alkynyl group, an optionally substituted C 3-12 cycloalkyl group, an optionally substituted 4- to 10-membered heterocyclic group, an optionally substituted C 6-10 aryl group, and an optionally substituted 5- to 10-membered heteroaryl group, wherein the optional substitution means that the hydrogen on the group to be substituted is unsubstituted or one or more substitutable sites of the group to be substituted are independently substituted by R 17 , and R 17 is, independently at each occurrence, hydrogen, halogen, hydroxy, mercapto, amino, cyano, carboxy, oxo, sulfoxy, -R 11 , -OR 11 , -SR 11 , -NH(R 11 ), -N(R 11 )(R 11 ), [Chemical formula] selected from, and R 11 is, independently at each occurrence, hydrogen, halogen, hydroxy, mercapto, amino, cyano, oxo, C 1-6 alkyl, trifluoromethyl, C 3-6 cycloalkyl, 4- to 6-membered heterocycloalkyl, aryl, a C optionally substituted by one or more selected from 5- to 6-membered heteroaryl 1-6 alkyl, C 1-6 alkoxy, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6Cycloalkyl, C 3-6 Selected from cycloalkoxy, 4- to 10-membered heterocyclic group, aryl, 5- to 6-membered heteroaryl, D is, in each occurrence, independently, one selected from a bond, -CH2-, -C(=O)-, -NH-, -N(CH3)-, -O-, -S-, f is, in each occurrence, independently, selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8, m = 0 or 1, and when m = 1, A may be selected from -N(R 12 )-, -CH(R 12 )-, or -CH(NHR 12 )-, R 12 is one selected from hydrogen, hydroxy group, halogen, C 1-6 alkyl group, C 3-6 cycloalkyl group, 4- to 6-membered heterocyclic group, and when m = 0, A does not exist, B is selected from N or CH, n = 0 or 1, and when n = 1, E is selected from -NR 13 - or -C(R 13 )R 13 -, R 13 is, in each occurrence, independently, one selected from hydrogen, hydroxy, halogen, C 1-6 alkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocyclic group, and when n = 0, E does not exist, At least one of A, B, and E is an N atom according to their respective definitions, o = 0, 1, 2 or 3, K may be selected from -CH2-, -C(=O)-, -CHF-, -CH(-OH)-, -CF2-, -CHD-, -CD(-OH)-, -CDF-, -CD2-, or -CH(R 18 )-, R 18 is, in each occurrence, independently, one selected from halogen, mercapto, nitro, cyano, amino, C 1-6 alkyl, C 3-6 cycloalkyl, 4- to 6-membered heterocyclic group, The H ring is C 6-10It is one selected from an aryl ring and a 5- to 10-membered heteroaryl ring, and the aryl ring or heteroaryl ring has one or more Rs 15 and may be independently substituted by 15 , and R 15 is, in each occurrence, independently hydrogen, halogen, hydroxy, mercapto, amino, cyano, optionally substituted -R 14 , optionally substituted -OR 14 , optionally substituted -NHR 14 , optionally substituted -N(R 14 )(R 14 ), and is selected from the group consisting of, provided that R 14 is, in each occurrence, independently C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group, aryl, a 5- to 6-membered heteroaryl, and the term "optionally substituted" means that the hydrogen on the group to be substituted is unsubstituted or one or more substitutable sites of the group to be substituted are independently substituted by a substituent selected from halogen, hydroxy group, mercapto group, amino group, cyano group, C 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3-6 cycloalkyl group, a 4- to 6-membered heterocyclic group, aryl group, a 5- to 6-membered heteroaryl group.

[0012] In a preferred embodiment of the present invention, the compound represented by formula (I) is represented by the structure represented by formula (II)B,

Chemical formula

[0013] In a more preferred embodiment, R 1 , R 2 are, in each occurrence, independently hydrogen, halogen, hydroxyl, mercapto, amino, cyano, C 1-6 alkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group, -OR 3, -NH(R 3 ), -N(R 3 )(R 4 ), and is one selected therefrom, and R 3 , R 4 is, independently at each occurrence, selected from C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group, aryl, and a 5- to 6-membered heteroaryl. Even more preferably, R 1 , R 2 is, independently at each occurrence, one selected from hydrogen, halogen, hydroxy, amino, methyl, methylamino, and dimethylamino, and most preferably hydrogen.

[0014] In some preferred embodiments of the present invention, preferably, X is selected from one of CH, C(OH), and N, and most preferably N.

[0015] Preferably, Y is —(chemical bond), -H, -OH, -NH2, halogen, -O-, -S-, -CO-, -C(R 6 )F-, -CF2-, -SO-, -SO2-, -(CH2) p N(R 6 ), -N(R 6 )(CH2) p -, -S(O)N(R 6 ), -S(O)2N(R 6 ), -N(R 6 )SO-, -N(R 6 )S(O)2-, -C(O)N(R 6 ), -N(R 6 )C(O)-, -CH(R 6 )-, and is one selected therefrom, provided that p = 0, 1, 2, or 3, and R 6 is one selected from hydrogen, C 1-6 alkyl, C 3-6 cycloalkyl, and a 4- to 6-membered heterocyclic group. When Y is one selected from -H, -OH, -NH2, and halogen, G 4 does not exist.

[0016] More preferably, Y is one selected from —(chemical bond), -H, -OH, -NH2, -NH-, -O-, -S-, -CO-, -CHF-, -CF2-, -SO-, -SO2-, -(CH2) p NH-, -N(CH3)-, -S(O)NH-, -S(O)2NH-, -NHSO-, -NHS(O)2-, -C(O)NH-, -NHC(O)-, -CH2-, and p = 1, 2 or 3.

[0017] Even more preferably, Y is one selected from —(chemical bond), -H, -OH, -NH2, -NH-, -CH2NH-, -(CH2)2NH-, -N(CH3)-, -O-, -S-.

[0018] Even more preferably, Y is selected from —(chemical bond), -OH, -NH2, -NH-, -O-, -S-. Most preferably, Y is selected from —(chemical bond), -NH-.

[0019] In some preferred embodiments of the present invention, Z is —(chemical bond), -O-, -S-, -CO-, -N(R 7 )-, -S(O)N(R 7 )-, -S(O)2N(R 7 )-, -N(R 7 )SO-, -N(R 7 )S(O)2-, -C(O)N(R 7 )-, -N(R 7 )C(O)-, -N(R 7 )C(O)N(R 7 )-, -CH(R 7 )-, -CH(OH)-, -CH(CF3)-, -CH(NHR 7 )-, -C=N(R 7 )-, -SO-, -SO2-, -CF(R 7 )-, CF2,

Chemical formula

[0020] More preferably, Z is —(chemical bond), -O-, -S-, -CO-, -NH-, -S(O)2NH-, -NHS(O)2-, -C(O)NH-, -NHC(O)-, -NHC(O)NH-, -CH2-, -CH(OH)-, -CH(CF3)-, -CH(NH2)-, -C=N(CH3)-, -C=N(OCH3)-, -SO-, -SO2-, -CHF-, CF2,

Chemical formula

[0021] Most preferably, Z is —(chemical bond), -O-, -CO-, -NH-, -C(O)NH-, -CH2-, -CH(OH)-, -CH(CF3)-, -CH(NH2)-, -C=N(CH3)-, -C=N(OCH3)-, -SO2-, -CHF-, CF2,

Chemical formula

[0022] Even more preferably, Z is selected from -CO- and -SO2-, and most preferably, Z is selected from -CO-.

[0023] In some preferred embodiments of the present invention, G 1 is, in each occurrence, independently selected from one of hydrogen, halogen, hydroxy, mercapto, amino, cyano, and methyl, and more preferably, G 1 is selected from hydrogen.

[0024] In some preferred embodiments of the present invention, G 2 is halogen, hydroxyl, mercapto, amino, cyano, -R 8 , -O(R 8 ), -S(R 8 ), -NH(R8 ), -N(R 8 )(R 8 ), and is selected from one kind, provided that R 8 is, in each occurrence independently, C 1-6 alkyl, C 3-6 cycloalkyl, or a 4- to 6-membered heterocyclic group, and more preferably, G 2 is selected from one kind of halogen, hydroxyl, mercapto, amino, cyano, -CH3, cyclopropyl, -OCH3, -SCH3, -NHCH3, -N(CH3)(CH3), -NH(CH3), and more preferably, G 2 is selected from one kind of fluorine, hydroxyl, amino, and most preferably, G 2 is selected from hydroxyl and amino.

[0025] In some preferred embodiments of the present invention, G 3 is selected from one kind of optionally substituted C 6-10 aryl, optionally substituted 5- to 10-membered heteroaryl, and optionally substituted 4- to 10-membered heterocyclic group, provided that the heteroatoms in the 5- to 10-membered heteroaryl are N, O, S, the number of heteroatoms is 1, 2, 3, or 4, and the term "optionally substituted" means that the hydrogen on the group to be substituted is unsubstituted or one or more substitutable sites of the group to be substituted are independently substituted by R 16 .

[0026] In a more preferred embodiment of the present invention, G 3 is selected from one kind of optionally substituted C 6-10 aryl, optionally substituted 5- to 10-membered heteroaryl, and optionally substituted 4- to 10-membered heterocyclic group, and the C 6-10 aryl, 5- to 10-membered heteroaryl, or 4- to 10-membered heterocyclic group is:

Chemical formula

[0027] More preferably, R 9 , R 10is, independently at each occurrence, hydrogen, halogen, hydroxyl, mercapto, amino, methylamino, dimethylamino, methoxy, -C(O)CH3, -COOC(CH3)3, -COOCH3, cyano, C 1-6 alkyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group, aryl, a 5- to 6-membered heteroaryl, optionally substituted by one or more of C 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3-10 cycloalkyl group, a 4- to 10-membered heterocyclic group, aryl group, a 5- to 10-membered heteroaryl group, preferably, R 9 , R 10 is, independently at each occurrence, hydrogen, halogen, hydroxyl, amino, cyano, C 1-3 alkyl, C 3-6 cycloalkyl, a 4- to 5-membered heterocyclic group, methylamino, dimethylamino, methoxy, -C(O)CH3, -COOC(CH3)3, -COOCH3, optionally substituted by one or more of C 1-6 alkyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 3-6 cycloalkyl group, a 4- to 10-membered heterocyclic group, phenyl, a 5- to 10-membered heteroaryl group, is selected from.

[0028] In a more preferred embodiment of the present invention, G 3 is

Chemical formula

[0029] In a more preferred embodiment of the present invention, R 16is independently selected from hydrogen, deuterium, -OCH3, -OCH2CH3, -SCH3, -SCH2CH3, -S-C(CH3)3, -O-C(CH3)3, -O-CH(CH3)2, -O-CH2CH(CH3)2, -O-CH(CH2CH3)2, -S-CH(CH3)2, -S-CH2CH(CH3)2, -S-CH(CH2CH3)2 at each occurrence.

[0030] In some preferred embodiments of the present invention, G 4 is a group selected from -SO2(CH3), optionally substituted C 1-6 alkyl, optionally substituted C 3-12 cycloalkyl, optionally substituted 4- to 10-membered heterocyclic group, optionally substituted C 6-10 aryl, optionally substituted 5- to 10-membered heteroaryl, where the optional substitution means that the hydrogen on the group to be substituted is unsubstituted or one or more substitutable sites of the group to be substituted are independently substituted with R 17 , and R 17 is independently selected from hydrogen, halogen, hydroxy, mercapto, amino, cyano, carboxy, oxo, -R 11 , -OR 11 , -SR 11 , -NH(R 11 ), -N(R 11 )(R 11 ) at each occurrence,

Chemical formula

[0031] In a more preferred embodiment of the present invention, G 4 is one selected from the group consisting of:

Chemical formula

Chemical formula

Chemical formula

[0032] Most preferably, G 4 is

Chemical formula

[0033] Preferably, R 17 is, in each occurrence independently, hydrogen, halogen, hydroxyl, mercapto, amino, cyano, carboxyl, oxo, sulfoxide, sulfone, -R 11 , -OR 11 , -SR 11 , -NH(R 11 ), -N(R 11 )(R 11 ),

Chemical formula

[0034] Preferably, R 11is, independently at each occurrence, hydrogen, hydroxy, cyano, amino, trifluoromethyl, difluoromethyl, trifluoroethyl, difluoroethyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, methoxy, ethoxy, n-propoxy, isopropoxy, cyclopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group, C 6-8 aryl, or a 5- to 6-membered heteroaryl, and is one selected therefrom.

[0035] Most preferably, R 17 is

Chemical formula

[0036] In some preferred embodiments of the present invention, A is absent.

[0037] In some preferred embodiments of the present invention, B is selected from N.

[0038] In some preferred embodiments of the present invention, n = 1, and E is selected from -CH(R 13 )- or -N(R 13 )-, and R 13 is one selected from hydrogen, hydroxyl, halogen, and methyl. More preferably, n = 1, and E is selected from -CH2- or -NH-, and most preferably, n = 1, and E is selected from -CH2-.

[0039] In some preferred embodiments of the present invention, o = 0, 1, or 2, and more preferably o = 1.

[0040] In some preferred embodiments of the present invention, K may be selected from -CH2-, -C(=O)-, -CHF-, -CH(-OH)-, -CF2-, -CHD-, -CD(-OH)-, -CDF-, -CD2-, more preferably, K may be selected from -CH2-, -C(=O)-, -CF2-, -CHD-, -CHF-, -CD2-, most preferably, K may be selected from -CH2-, -C(=O)-, -CF2-, -CD2-, -CHD-, and most preferably, K may be selected from -CD2-.

[0041] In some preferred embodiments of the present invention, the H ring is one selected from a benzene ring, a 5- to 6-membered heteroaryl ring, a 6-membered heteroaryl ring condensed to a 5-membered ring portion, and a 5-membered heteroaryl ring condensed to a 6-membered ring portion. The benzene ring, 5- to 6-membered heteroaryl ring, 6-membered heteroaryl ring condensed to a 5-membered ring portion, and 5-membered heteroaryl ring condensed to a 6-membered ring portion may be independently substituted with one or more Rs 15 provided that R 15 is selected from hydrogen, halogen, hydroxy, mercapto, amino, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, a 4- to 6-membered heterocyclic group, aryl, and 5- to 6-membered heteroaryl.

[0042] In a preferred embodiment, the H ring is independently a benzene ring which may be substituted with one or more Rs 15 provided that R

[0043] In a preferred embodiment, the H ring is a 5- to 6-membered heteroaryl ring selected from the following:

Chemical formula

[0044] In a preferred embodiment, the H ring is a heteroaryl ring of a 6-membered ring fused to a 5-membered ring portion, [Chemical formula] and is selected from the following. The heteroaryl ring of the 6-membered ring fused to the 5-membered ring portion may be independently substituted with one or more Rs 15 .

[0045] In a preferred embodiment, the H ring is a heteroaryl ring of a 5-membered ring fused to a 6-membered ring portion, [Chemical formula] and is selected from the following. The heteroaryl ring of the 5-membered ring fused to the 6-membered ring portion may be independently substituted with one or more Rs 15 .

[0046] More preferably, R 15 is selected from hydrogen, halogen, hydroxyl, mercapto, amino, cyano, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 4-6 membered heterocyclic group, aryl, 5-6 membered heteroaryl.

[0047] In some preferred embodiments of the present invention, [Chemical formula] is [Chemical formula] .

[0048] In some preferred embodiments of the present invention, [Chemical formula] is [Chemical formula] is as follows.

[0049] In some preferred embodiments of the present invention, G in formula (I) or formula (II)B 3 is

Chemical formula

[0050] Preferably, R 16 is selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, -R 9 , -OR 9 , -SR 9 , -NH(R 9 ), and -N(R 9 )(R 10 ).

[0051] R 9 , R 10 are, independently in each occurrence, selected from C 1-6 alkyl and C 3-6 cycloalkyl, or R 9 and R 10 together with the N atom to which they are attached form a 4- to 6-membered heterocyclic group.

[0052] In some preferred embodiments of the present invention, G in formula (I) or formula (II)B 4 is

Chemical formula

[0053] Preferably, R 11 is selected from C 6-8 aryl and 5- to 6-membered heteroaryl, and the aryl or heteroaryl is optionally substituted with one or more of halogen, C 1-6 alkyl, and trifluoromethyl.

[0054] The complex ring group or heteroaryl group each independently contains 1, 2, or 3 heteroatoms selected from N, O, and S.

[0055] In some preferred embodiments of the present invention, the compound represented by formula (I) is a compound represented by formula (III),

Chemical formula

[0056] wherein R 16 is selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, -R 9 , -OR 9 , -SR 9 , -NH(R 9 ), and -N(R 9 )(R 10 ), R 9 , R 10 is independently selected from C 1-6 alkyl and C 3-6 cycloalkyl in each occurrence, or R 9 and R 10 form a 4- to 6-membered heterocyclic group together with the N atom to which they are attached, R 11 is selected from C 6-8 aryl and 5- to 6-membered heteroaryl, and the aryl or heteroaryl is optionally substituted with one or more of halogen, C 1-6 alkyl, and trifluoromethyl, The complex ring group or heteroaryl group each independently contains 1, 2, or 3 heteroatoms selected from N, O, and S.

[0057] In some more preferred embodiments of the present invention, R 16 is selected from hydrogen, deuterium, -R 9 , -OR 9 , -SR 9 , and -N(R 9 )(R 10 ), provided that R9 and R 10 is as defined in formula (III).

[0058] In some further preferred embodiments of the present invention, R 16 is selected from hydrogen, deuterium, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCH2CH(CH3)2, -OC(CH3)3, -OCH(CH2CH3)2, -O-cyclopropyl, -O-cyclobutyl, -O-cyclopentyl, -O-cyclohexyl, -SCH3, -SCH2CH3, -SCH(CH3)2, -SCH2CH(CH3)2, -SC(CH3)3, -SCH(CH2CH3)2, -S-cyclopropyl, -S-cyclobutyl, -S-cyclopentyl, -S-cyclohexyl, azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, and 3-azabicyclo[3.1.0]hex-3-yl.

[0059] In some further preferred embodiments of the present invention, R 11 is selected from phenyl, pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyridazinyl, and pyrazinyl, and the phenyl, pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyridazinyl, or pyrazinyl is optionally substituted with one or more of fluorine, methyl, and trifluoromethyl.

[0060] In some further preferred embodiments of the present invention, R 11is selected from phenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2-(trifluoromethyl)phenyl, 3-(trifluoromethyl)phenyl, 4-(trifluoromethyl)phenyl, thiophen-2-yl, thiophen-3-yl, 1-methyl-1H-pyrazol-3-yl, 1-methyl-1H-pyrazol-4-yl, 1-methyl-1H-pyrazol-5-yl, isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl, thiazol-2-yl, thiazol-4-yl, thiazol-5-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, and pyrimidin-5-yl.

[0061] In some further preferred embodiments of the present invention, the compound represented by formula (I) is a compound represented by formula (III)A or formula (III)B:

Chemical formula

[0062] wherein, R 11 and R 16 are as defined in formula (III).

[0063] The compounds of the present invention, and their stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotopically labeled analogs are selected from the following compounds.

Table 1-1

Table 1-2

Table 1-3

Table 1-4

Table 1-5

Table 1-6

Table 1-7

Table 1-8

Table 1-9

Table 1-10

Table 1-11

Table 1-12

Table 1-13

Table 1-14

Table 1-15

Table 1-16

Table 1-17

Table 1-18

Table 1-19

Table 1-20

Table 1-21

Table 1-22

Table 1-23

[0064] The present invention also aims to provide a method for preparing a compound represented by the general formula (I), and its stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates or isotope-labeled analogs.

[0065] In this method, for example, it can be prepared using the method shown in Scheme 1. First, LG a (leaving group a) of the left side chain of the G3-Z structure is bonded to G 4 -YH, and then LG c (leaving group c) of the cyclic structure is bonded to LG b (leaving group b) of the right side chain of the G3-Z structure, whereby the target compound can be synthesized.

[0066] Scheme 1 is as follows.

Chemical formula

[0067] In this method, for example, it can be prepared using the method shown in Scheme 2. First, LG c (leaving group c) of the cyclic structure is bonded to LG b (leaving group b) of the right side chain of the G3-Z structure, and then LG a (leaving group a) of the left side chain of the G3-Z structure is bonded to G 4 -YH, whereby the target compound can be synthesized.

[0068] Scheme 2 is as follows.

Chemical formula

[0069] In addition, in the above preparation method, the definitions of the respective substituents in the compounds represented by the respective formulas are as described above.

[0070] The present invention also provides a pharmaceutical composition comprising a compound of the present invention, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, solvate or isotope-labeled analog thereof, and a pharmaceutically acceptable excipient.

[0071] The present invention also aims to provide the use of a compound of the present invention, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, solvate or isotope-labeled analog thereof, in the preparation of a medicament for the prevention and / or treatment of diseases mediated by PRMT5. In some embodiments, the disease mediated by PRMT5 is a cancer or tumor-related disease.

[0072] The present invention aims to provide a method for the prevention and / or treatment of diseases mediated by PRMT5. The method includes administering to a patient a compound of the present invention, or a stereoisomer, geometric isomer, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, solvate, isotope-labeled analog, or the pharmaceutical composition of the present invention, in a therapeutically effective amount.

[0073] In some embodiments, the disease mediated by PRMT5 is a cancer or tumor-related disease. In some embodiments, the cancer or tumor-related disease is preferably lymphoma.

[0074] When the compound of the present invention, or a pharmaceutically acceptable salt thereof, is administered in combination with other anti-cancer agents or immune checkpoint inhibitors for the treatment of cancer or tumors, the compound of the present invention, or a pharmaceutically acceptable salt thereof, can provide an enhanced anti-cancer effect.

Advantages of the Invention

[0075] Advantageous effects of the present invention: The present invention designed a compound with a new structure, which provides a new direction for the development of drugs of PRMT5 inhibitors. As revealed by the research on the inhibitory activity of in vitro enzyme activity, these compounds have a strong inhibitory effect on the PRMT5 enzyme, and show excellent tumor suppression effect in the subcutaneous xenograft NOD / SCID female mouse test of the cell line of human B-cell non-Hodgkin lymphoma Z-138, and show excellent metabolic characteristics in the in vivo pharmacokinetic test of mice. Therefore, it is shown that they are promising compounds for the treatment of diseases mediated by PRMT5. Furthermore, the present invention also studied a specific synthesis method which has a simple process, convenient operation and is advantageous for large-scale industrial production and application.

Mode for Carrying Out the Invention

[0076] [Definition of Terms] Unless otherwise specified, the term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, which is a straight-chain or branched-chain group containing 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms (i.e., C 1-10 alkyl group), more preferably 1 to 8 carbon atoms (C 1-8 alkyl group), still more preferably 1 to 6 carbon atoms (i.e., C 1-6 alkyl group). For example, the "C 1-6 alkyl group" refers to an alkyl group having 1 to 6 carbon atoms (specifically, 1, 2, 3, 4, 5, or 6) on the carbon chain. Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, n-octyl, etc.

[0077] Unless otherwise specified, the term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group having a specific number of carbon atoms, preferably 3 to 12 carbon atoms (i.e., C 3-12including a cycloalkyl group), more preferably 3 to 10 carbon atoms (C 3-10 including a cycloalkyl group), even more preferably 3 to 6 carbon atoms (C 3-6 cycloalkyl group), 4 to 6 carbon atoms (C 4-6 cycloalkyl group), 5 to 6 carbon atoms (C 5-6 cycloalkyl group). Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, etc.

[0078] Unless otherwise specified, the term "alkoxy group" refers to an -O-alkyl group, and the definition of the alkyl group is the same as above, i.e., including 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 8 carbon atoms, even more preferably 1 to 6 carbon atoms (specifically 1, 2, 3, 4, 5, or 6). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, 1-methylpropoxy, 2-methylpropoxy, tert-butoxy, pentyloxy, 1-methylbutoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, 2,2-dimethylpropoxy, 1-ethylpropoxy, etc.

[0079] Unless otherwise specified, the term "halogen" or "halogenated" refers to F, Cl, Br, I. The term "halogenated alkyl group" refers to the substitution of one, two or more hydrogen atoms, or all hydrogen atoms, in the alkyl group defined as above with halogen. Representative examples of halogenated alkyl groups include CCl3, CF3, CHCl2, CH2Cl, CH2Br, CH2I, CH2CF3, CF2CF3, etc.

[0080] Unless otherwise specified, the term "heterocyclic group" refers to a saturated or partially unsaturated monocyclic, bicyclic, or polycyclic hydrocarbon substituent that has a non-aromatic structure, contains 3 to 20 ring atoms, of which one, two, three, or more ring atoms are selected from N, O, or S, and the other ring atoms are C. Preferably, it contains 3 to 12 ring atoms, more preferably 3 to 10 ring atoms, or 3 to 8 ring atoms, or 3 to 6 ring atoms, or 4 to 6 ring atoms, or 5 to 6 ring atoms. The heteroatoms are preferably 1 to 4, more preferably 1 to 3 (i.e., one, two, or three). Examples of monocyclic heterocyclic groups include pyrrolidyl, imidazolidinyl, tetrahydrofuran, dihydropyrrole, piperidinyl, piperazinyl, pyranyl, and the like. Polycyclic heterocyclic groups include spiro ring, fused ring, and bridged fused ring heterocyclic groups.

[0081] Unless otherwise specified, the term "carbocyclic group" or "carbocycle" refers to a cyclic non-aromatic hydrocarbon group (a "C 3-14 carbocyclic group") having 3 to 14 ring-forming carbon atoms and having no heteroatoms in the non-aromatic ring system. In some embodiments, the carbocyclic group has 3 to 12 ring-forming carbon atoms (a "C 3-12 carbocyclic group"), or 4 to 12 ring-forming carbon atoms (a "C 4-12 carbocyclic group"), or 3 to 10 ring-forming carbon atoms (a "C 3-10 carbocyclic group"). In some embodiments, the carbocyclic group has 3 to 8 ring-forming carbon atoms (a "C 3-8 carbocyclic group"). In some embodiments, the carbocyclic group has 3 to 7 ring-forming carbon atoms (a "C 3-7 carbocyclic group"). In some embodiments, the carbocyclic group has 4 to 6 ring-forming carbon atoms (a "C 4-6 carbocyclic group"). In some embodiments, the carbocyclic group has 5 to 10 ring-forming carbon atoms (a "C 5-10 carbocyclic group"), or 5 to 7 ring-forming carbon atoms (a "C 5-7 carbocyclic group"). Exemplary C 3-6Examples of the carbocyclic group include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentene (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexanedienyl (C6), etc. Exemplary C 3-8 Examples of the carbocyclic group include the above C 3-6 In addition to the carbocyclic groups, cycloheptyl (C7), cycloheptene (C7), cycloheptanedienyl (C7), cycloheptatrieneyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptane (C7), bicyclo[2.2.2]octane (C8), etc. are included, but are not limited thereto. Exemplary C 3-10 Examples of the carbocyclic group include the above C 3-8 In addition to the carbocyclic groups, cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C 10 ), cyclodecenyl (C 10 ), octahydro-1H-indenyl (C9), decahydronaphthyl (C 10 ), spiro[4.5]decyl (C 10 ), etc. are included, but are not limited thereto. As described in the above examples, in some embodiments, the carbocyclic group is monocyclic ("monocyclic carbocyclic group"), or a ring system of fused (fused ring group), bridged (bridged fused ring group), or spiro-fused (spiro ring group), for example bicyclic ("bicyclic carbocyclic group"), and may be saturated or partially unsaturated. The "carbocyclic group" further includes a ring system in which the ring of the carbocyclic group defined as above is fused with one or more aryl groups or heteroaryl groups, provided that the bonding point is on the ring of the carbocyclic group, and in such a case, the number of carbons continues to indicate the number of carbons in the carbocyclic system. In some embodiments, each example of the carbocyclic group may be optionally substituted, such as unsubstituted ("unsubstituted carbocyclic group") or substituted with one or more substituents ("substituted carbocyclic group"). In some embodiments, the carbocyclic group is an unsubstituted C 3-10 carbocyclic group. In some embodiments, the carbocyclic group is a substituted C 3-10 carbocyclic group.

[0082] Unless otherwise specified, the term "aryl" refers to monocyclic, bicyclic and tricyclic aromatic carbocyclic systems containing 6 to 16 carbon atoms, or 6 to 14 carbon atoms, or 6 to 12 carbon atoms, or 6 to 10 carbon atoms, preferably 6 to 10 carbon atoms. The term "aryl" can be used interchangeably with the term "aromatic ring". Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, phenanthrene, pyrenyl, etc.

[0083] Unless otherwise specified, the term "heteroaryl group" refers to aromatic monocyclic or polycyclic ring systems having a 5- to 12-membered structure, or preferably a 5- to 10-membered structure, a 5- to 8-membered structure, more preferably a 5- to 6-membered structure. However, among the ring atoms, one, two, three or more are heteroatoms and the remaining atoms are carbon. The heteroatoms are independently selected from O, N, or S, and the number of heteroatoms is preferably 1, 2 or 3. Examples of heteroaryl groups include, but are not limited to, furyl, thienyl, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, thiadiazolyl, triazinyl, phthalazinyl, quinolyl, isoquinolyl, pteridinyl, purinyl, indolyl, isoindolyl, indazolyl, benzofuranyl, benzothienyl, benzopyridinyl, benzopyrimidinyl, benzopyrazinyl, benzimidazolyl, benzophthalazinyl, pyrrolo[2,3-b]pyridinyl, imidazolyl[1,2-a]pyridinyl, pyrazolyl[1,5-a]pyridinyl, pyrazolyl[1,5-a]pyrimidinyl, imidazolyl[1,2-b]pyridazinyl, [1,2,4]triazolyl[4,3-b]pyridazinyl, [1,2,4]triazolyl[1,5-a]pyrimidinyl, [1,2,4]triazolyl[1,5-a]pyridinyl, etc.

[0084] Unless otherwise specified, the terms "pharmaceutically acceptable salt", "medicinal salt", or "medicinally acceptable salt" refer to salts that are suitable for contact with mammalian, particularly human, tissues within the scope of reasonable medical judgment, having no excessive toxicity, irritation, allergic reaction, etc., and having an appropriate benefit / risk ratio. For example, pharmaceutically acceptable salts of amines, carboxylic acids, and other compounds are well-known to those skilled in the art. Said salts can be prepared in-situ during the final separation and purification of the compounds of the present invention, or can be prepared by reacting the free base or free acid alone with an appropriate reagent, as outlined below. For example, the free base functionality can react with an appropriate acid.

[0085] Unless otherwise specified, the term "solvate" refers to the physical association of a compound of the present invention with one or more solvent molecules (whether organic or inorganic). The physical association includes hydrogen bonding. In some cases, for example, when one or more solvent molecules are incorporated into the lattice of a crystalline solid, the solvate can be separated. The solvent molecules in the solvate can exist in a regular arrangement and / or a disordered arrangement. The solvate can contain stoichiometric or non-stoichiometric amounts of solvent molecules. "Solvate" includes solvates separable from the solution phase. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Methods of solvation are known in the art.

[0086] Unless otherwise specified, the term "isotopically labeled analog" refers to a molecule labeled with an isotope in the compounds of Formulas I to II, thereby providing an isotopically labeled analog that may have improved pharmacological activity. Isotopes commonly used as isotopic labels are hydrogen isotopes 2 H and 3 H, carbon isotopes 11 C, 13 C and 13 C, chlorine isotope 35 Cl and 37 Cl, fluorine isotope 18 F, iodine isotope 123 I and125 I. Nitrogen isotopes 13 N and 15 N, oxygen isotopes 15 O, 17 O and 18 O, and sulfur isotope 35 S. These isotope-labeled compounds can be used to study the distribution of pharmaceutical molecules in tissues. In particular, deuterium (D, or 2 H), 3 H and carbon 13 C are widely used because they are easy to label and detect. Substitution with heavy isotopes such as deuterium ( 2 H) can enhance metabolic stability, achieve the purpose of extending the half-life and reducing the dose, and provide the superiority of therapeutic effect. Generally, isotope-labeled compounds are synthesized from labeled starting materials by known synthetic techniques in the same manner as the synthesis of non-isotope-labeled compounds.

[0087] Unless otherwise specified, the term "prodrug" refers to a drug that is converted into a parent drug in vivo. Prodrugs are usually useful because they may be easier to administer than the parent drug in some cases. For example, a prodrug can be made bioavailable to the body by oral administration while the parent drug cannot. A prodrug can have higher solubility in a pharmaceutical composition than the parent drug. As an example of a prodrug, it may be, but is not limited to, any compound of formula (I) administered as an ester ("prodrug") to facilitate movement across cell membranes. However, water solubility is harmful to mobility, but becomes beneficial after entering the cell and is then metabolized and hydrolyzed to a carboxylic acid, i.e., the active form. Another example of a prodrug is a short peptide (polyamino acid) bonded to an acid group. The peptide is metabolized to express the active moiety.

[0088] Unless otherwise specified, the term "stereoisomer" refers to compounds that have the same chemical structure but differ in the spatial arrangement of atoms or groups. Stereoisomers include enantiomers, diastereoisomers, conformational isomers (rotational isomers), geometric (cis / trans) isomers, atropisomers, etc. Any mixture of obtained stereoisomers may be separated into neat or substantially neat geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization methods.

[0089] Unless otherwise specified, the term "tautomer" refers to structural isomers of different energies that can interconvert via a low energy barrier. When tautomerism is possible (for example, when in solution), the chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions by proton transfer such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions by rearrangement of some bonding electrons.

[0090] Unless otherwise expressly stated, the structures shown in the present invention also include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure; for example, the R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, as well as (Z) and (E) conformational isomers. Accordingly, individual stereochemical isomers, enantiomers, diastereomers, or mixtures of geometric isomers (or conformational isomers) are within the scope disclosed in the present invention.

[0091] Unless otherwise specified, the term "optional substitution" refers to the hydrogen at the substitutable site of the base being unsubstituted or substituted with one or more substituents. The substituents include halogen, hydroxy group, mercapto group, cyano group, nitro group, amino group, azide group, oxo group, carboxyl group, C 2-6 alkenyl group, C 2-6 alkynyl group, C 1-6 alkyl group, C 1-6 alkoxy group, C 3-10 cycloalkyl group, C 3-10 cycloalkylsulfonyl group, 3- to 10-membered heterocycloalkyl group, C 6-14 aryl group, or a 5- to 10-membered heteroaryl ring group, preferably selected from the group consisting of. However, the C 2-6 alkenyl group, C 2-6 alkynyl group, C 1-6 alkyl group, C 3-10 cycloalkyl group, C 3-10 cycloalkylsulfonyl group, 3- to 10-membered heterocycloalkyl group, C 6-14 aryl group, or a 5- to 10-membered heteroaryl ring group may be optionally substituted with one or more selected from halogen, hydroxy group, amino group, cyano group, C 1-6 alkyl group, or C 1-6 alkoxy group. The oxo group refers to the formation of a double bond by replacing two Hs at the same substitution site with the same O.

[0092] Hereinafter, the present invention will be further described based on specific examples. It should be understood that these examples are not intended to limit the scope of the present invention, but are used only for the purpose of explaining the present invention. Experimental methods for which specific conditions are not specified in the following examples usually follow normal conditions or the recommended conditions of the manufacturer. Unless otherwise defined specifically, all technical terms and scientific terms used in this text have the same meaning as those well-known to those skilled in the art. Also, any methods and materials similar or equivalent to the described content can be applied to the method of the present invention. The more preferred implementation methods and materials shown in this text are used only by way of illustration.

[0093] The compound structure of the present invention is determined by nuclear magnetic resonance (NMR) or / and liquid chromatography / mass fraction analysis (LC-MS) or / and high performance liquid chromatography (HPLC). The instrument used for NMR measurement is Bruker AVANCE NEO 400 MHz, the instrument used for LC-MS is LCMS WATERS ACQUITY UPLC H-Class PLUS or / and SQD 2, and the instrument used for HPLC is WATERS ACQUITY UPLC and / or Agilent 1260.

[0094] The starting materials in the examples of the present invention are known and can be obtained commercially, or may be synthesized by or according to methods known in the art.

[0095] [Preparation of Intermediate] Production Example 1: Preparation of Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate [Chemical Formula]

[0096] Methyl 2,6-dichloropyrimidine-4-carboxylate (6 g, 28.98 mmol), 1-acetylpiperidin-4-amine hydrochloride (5.7 g, 31.88 mmol) and DIPEA (14.98 g, 115.94 mmol) were dissolved in acetonitrile (50 mL). The mixture was stirred at room temperature (25 - 30 °C) for 2 hours. It was extracted three times with 100 mL of ethyl acetate each time. The ethyl acetate phases were combined, washed once with 50 mL of water, washed once with 50 mL of saturated brine, dried over anhydrous sodium sulfate for 10 minutes, filtered, and the crude product was separated and purified by column chromatography using DCM:MeOH = 100:1 to obtain the product (5.8 g, yield 64%). LC-MS (ESI) [M + H] + = 313.2.

[0097] Production Example 2: Preparation of trans-(6-chloropyrimidin-4-yl)(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone (This intermediate can be used, for example, as an intermediate for the compound of Example 1)

[0098] Step 1: Preparation of 6-chloropyrimidine-4-carbonyl chloride

Chemical formula

[0099] 4,6-Dichloropyrimidine (570 mg, 3.83 mmol, 1.0 eq) was dissolved in EA (18 mL), and oxalyl chloride (2.43 g, 19.13 mmol, 5.0 eq) and N,N-dimethylformamide (1.8 mL) were added. The reaction was carried out at 85 °C for 2 hours. After monitoring the completion of the reaction by TLC plate and LC-MS, the reaction solution was quickly dried on a rotary evaporator and sealed for the next step.

[0100] Step 2: Preparation of tert-butyl trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carboxylate

Chemical formula

[0101] tert-Butyl 7-oxa-3-azabicyclo[4.1.0]heptane-3-carboxylate (2.50 g, 12.56 mmol, 1.0 eq) was dissolved in i-PrOH (isopropanol, 63 mL), 1,2,3,4-tetrahydroisoquinoline (1.67 g, 12.56 mmol, 1.0 eq) was added, and the reaction was carried out at 85 °C for 18 h under nitrogen protection. After monitoring the completion of the reaction by TLC plate and LC-MS, the reaction solvent was dried by a rotary evaporator, water (200 mL) was added, and the mixture was extracted with dichloromethane (200 mL each, 3 times in total), dried over anhydrous sodium sulfate, suction filtered, and dried by a rotary evaporator. In this step, positional isomers including tert-butyl trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carboxylate and tert-butyl trans-3-(3,4-dihydroisoquinolin-2(1H)-yl)-4-hydroxypiperidine-1-carboxylate were present. The crude mixture (3.40 g, yield 81.5%) was separated and purified by chromatography (silica gel, ethyl acetate:petroleum ether = 15:85) 2 - 3 times to obtain the title compound (1.7 g, yield 41%).

[0102] LC-MS(ESI)[ M + H] + = 333.3, 1 H NMR (400 MHz, CDCl3) δ 7.20 - 7.08 (m, 3H), 7.05 - 6.98 (m, 1H), 4.59 - 4.17 (m, 2H), 3.94 (d, J = 14.6 Hz, 1H), 3.68 (d, J = 14.5 Hz, 2H), 3.54 (td, J = 10.0, 5.0 Hz, 1H), 3.03 (dt, J = 10.9, 5.3 Hz, 1H), 2.91 (t, J = 5.6 Hz, 2H), 2.80 - 2.47 (m, 4H), 1.82 (dd, J = 12.7, 2.5 Hz, 1H), 1.59 - 1.38 (m, 10H).

[0103] Step 3: Preparation of trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol

Chem.

[0104] tert-Butyl trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carboxylate (1.00 g, 3.01 mmol, 1.0 eq) was dissolved in DCM (dichloromethane) (15 mL) and TFA (trifluoroacetic acid) (3.75 mL). The reaction mixture was stirred at room temperature (20 - 25 °C) for 2 hours. After monitoring the completion of the reaction by TLC plate and LC-MS, the reaction solution was dried on a rotary evaporator, and 1,2-dichloroethane was added to remove the excess TFA by repeating 3 times. After drying, it was sealed and used directly in Step 3. LC-MS (ESI) [M + H] + = 233.2.

[0105] Step 4: Preparation of trans-(6-chloropyrimidin-4-yl)(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone

Chem.

[0106] trans-4-(3,4-Dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (570 mg, 3.23 mmol, 1.0 eq) was dissolved in DCM (dichloromethane) (8 mL) and TEA (triethylamine) (653 mg, 6.46 mmol, 2.0 eq). 6-Chloropyrimidine-4-carbonyl chloride was dissolved in dichloromethane (8 mL), and slowly added to the reaction solution of trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol under nitrogen protection. The reaction mixture was reacted in an ice bath for 2 hours, and then gradually warmed to room temperature (20 - 25 °C) and reacted for 1 hour. After monitoring the completion of the reaction by TLC plate and LC-MS, 100 ml of water was added to the reaction solution, and it was extracted with ethyl acetate and dried over anhydrous sodium sulfate. The organic phase was concentrated and separated and purified by reverse-phase HPLC (C18, 0.08% aqueous NH4HCO3 solution / acetonitrile) to obtain the title compound (900 mg, yield 56.3%). LC-MS(ESI)[ M + H] + = 373。

[0107] Production Example 3: 6-((1-Acetylpiperidin-4-yl)amino)-2-(benzo[d]thiazol-5-yl)pyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 9)

Chemical Structure

[0108] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (90 mg, 0.29 mmol, 1.0 equiv.), benzothiazole-5-boronic acid pinacol ester (113 mg, 0.43 mmol, 1.5 equiv.), 1,1'-bis-diphenylphosphinopalladium dichloride iron (21 mg, 0.03 mmol, 0.1 equiv.) and potassium phosphate (183 mg, 0.86 mmol, 3.0 equiv.) were dissolved in dioxane (2 mL) and water (0.5 mL), and the mixture was replaced with nitrogen three times. The reaction was carried out at 120 °C for 16 hours. After monitoring the completion of the reaction by LC-MS, the mixture was extracted three times with dichloromethane (5 mL), the aqueous phase was concentrated, dried on a rotary evaporator to remove the solvent, and the crude product (80 mg) of the title compound was obtained. LC-MS (ESI) [M + H] + = 398.20.

[0109] Production Example 4: 2-Phenyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 10)

[0110] Step 1: Preparation of methyl 2-chloro-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylate

Chemical formula

[0111] Methyl 2,6-dichloropyrimidine-4-carboxylate (500 mg, 2.41 mmol, 1.01 equiv.) and tetrahydro-2H-pyran-4-amine (240 mg, 2.37 mmol, 1 equiv.) were dissolved in acetonitrile (12 mL), and DIPEA (920 mg, 7.12 mmol, 3 equiv.) was added. The reaction mixture was stirred at 80 °C for 3 h. The completion of the reaction was monitored by LC-MS. Water was added to the reaction solution to quench it, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was separated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate = 0 - 15%) to give the title compound (500 mg, yield 82%). LC-MS (ESI) [M + H] + = 272.1.

[0112] Step 2: Preparation of 2-phenyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylic acid

Chemical Structure

[0113] Methyl 2-chloro-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylate (200 mg, 0.737 mmol, 1.0 equiv.), benzeneboronic acid (140 mg, 1.106 mmol, 1.5 equiv.) were dissolved in 1,4-dioxane (2.5 mL) and water (0.5 mL), and Pd(dppf)Cl2 (270 mg, 0.369 mmol, 0.5 equiv.) and potassium phosphate (470 mg, 2.211 mmol, 3 equiv.) were added. The reaction mixture was stirred at 100 °C for 8 hours. The completion of the reaction was monitored by LC-MS. Water was added to the reaction solution to quench it, and it was filtered through celite to remove insoluble solid impurities. The filtrate was extracted with ethyl acetate, the aqueous phase was directly concentrated, the obtained solid was added to methylene chloride and methanol to dissolve it, filtered, the filtrate was collected, and after concentrating the filtrate, the crude product (98 mg) of the title compound was used directly in the next step. LC-MS(ESI)[ M + H] + =300.2。

[0114] Production Example 5: 6-((1-Acetylpiperidin-4-yl)amino)-2-(benzo[d]oxazol-7-yl)pyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 13)

[0115] Step 1: Preparation of 7-bromopyrrolo[d]oxazole

Chemical Structure

[0116] 2-Amino-6-bromophenol (1.0 g, 5.319 mmol, 1 equiv.) was dissolved in trimethyl orthoformate (10 mL), and p-toluenesulfonic acid (100 mg, 0.581 mmol, 0.11 equiv.) was added. The reaction mixture was stirred at 80 °C for 2 h under nitrogen. The completion of the reaction was monitored by TLC. The reaction mixture was dried by rotary evaporator under reduced pressure, and the crude product was separated and purified by flash chromatography (silica gel, EA:PE = 1:20) to obtain the title compound (1.12 g, yield 98.9%). LC-MS (ESI) [M + H] + = 180.0。

[0117] Step 2: Preparation of 7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoxazole

Chemical formula

[0118] 7-Bromopyrrolo[d]oxazole (210 mg, 1.060 mmol, 1 equiv.), pinacol diborate (645 mg, 2.540 mmol, 2.4 equiv.), Pd(dppf)Cl2 (74 mg, 0.102 mmol, 0.1 equiv.) and potassium acetate (200 mg, 2.038 mmol, 1.92 equiv.) were weighed and added to a microwave vial, capped and purged under nitrogen conditions. 1,4-Dioxane (5 mL) was added. The reaction was heated to 100 °C for 2 h under a nitrogen atmosphere. The completion of the reaction was monitored by LC-MS. The reaction solution was filtered and dried by rotary evaporator, and the crude product was separated and purified by flash chromatography (silica gel, EA:PE = 1:20) to obtain the title compound (252 mg, yield 97%). LC-MS (ESI) [M+H] + = 246.2, 341.0; 11H NMR (400 MHz, CDCl3) δ 8.20 (s, 1H), 7.90 (d, J = 7.9 Hz, 1H), 7.83 (d, J = 7.4 Hz, 1H), 7.39 (t, J = 7.6 Hz, 1H), 1.42 (s, 12H).

[0119] Step 3: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(benzo[d]oxazol-7-yl)pyrimidine-4-carboxylate

Chemical Structure

[0120] 7-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]oxazole (100 mg, 0.408 mmol, 1 equiv.), methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (127 mg, 0.406 mmol, 1 equiv.), Pd(dppf)Cl2 (30 mg, 0.0410 mmol, 0.1 equiv.) and sodium carbonate (80 mg, 0.755 mmol, 1.86 equiv.) were weighed into a 50 mL single-neck flask, 1,4-dioxane (2 mL) and water (0.5 mL) were added, and the reaction system was purged with nitrogen. The reaction was heated at 90 °C for 1 hour. The completion of the reaction was monitored by LC-MS. Ethyl acetate and water were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (2 × 15 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and dried on a rotary evaporator. The crude product was separated and purified by flash chromatography (silica gel, MeOH:DCM = 1:25) to obtain the title compound (66 mg, yield 41.1%). LC-MS (ESI) [M + H] + = 396.20。

[0121] Step 4: Preparation of 6-((1-Acetylpiperidin-4-yl)amino)-2-(benzo[d]oxazol-7-yl)pyrimidine-4-carboxylic acid (Intermediate 3)

Chem.

[0122] Tributyltin oxide (180 mg, 0.302 mmol, 1.99 eq) was added to a suspension of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(benzo[d]oxazol-7-yl)pyrimidine-4-carboxylate (60 mg, 0.152 mmol, 1 eq) in toluene (5 mL), protected with nitrogen, and heated to reflux at 110 °C for 4 h. The completion of the reaction was monitored by TLC. Toluene was dried by rotary evaporator, and petroleum ether was added to form a pulp. After filtration, it was washed with petroleum ether, and the residual solvent was suction-dried by pump to obtain 75 mg of the crude title compound, which was directly used in the next step. LC-MS (ESI) [M + H] + = 382.0.

[0123] Production Example 6: tert-Butyl 4-((6-(3R,4R-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-phenylpyrimidin-4-yl)amino)piperidine-1-carboxylate (This intermediate can be used, for example, as an intermediate for the compound of Example 20)

[0124] Step 1: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate

Chem.

[0125] Methyl 2,6-dichloropyrimidine-4-carboxylate (1.086 g, 5.24 mmol, 1.05 equiv.) and tert-butyl 4-aminopiperidine-1-carboxylate (1 g, 4.99 mmol, 1.05 equiv.) were dissolved in acetonitrile (40 mL), and DIPEA (1.94 g, 14.97 mmol, 3 equiv.) was added. The reaction mixture was stirred at 80 °C for 3 h. The completion of the reaction was monitored by LC-MS. Water was added to the reaction solution to quench it, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was separated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate = 1:5) to give the title compound (1.5 g, yield 80%). LC-MS(ESI)[ M + H] + =371.0。

[0126] Step 2: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-phenylpyrimidine-4-carboxylate

Chemical Structure

[0127] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (300 mg, 0.809 mmol, 1.0 equiv.), benzeneboronic acid (148 mg, 1.213 mmol, 1.5 equiv.) were dissolved in 1,4-dioxane (8 mL) and water (2 mL), and Pd(dppf)Cl2 (118 mg, 0.162 mmol, 0.2 equiv.) and potassium phosphate (859 mg, 4.045 mmol, 5 equiv.) were added. The reaction mixture was stirred at 100 °C for 18 h. The completion of the reaction was monitored by LC-MS. The reaction solution was filtered to remove insoluble solid impurities. The filtrate was extracted with ethyl acetate, and the aqueous phase was concentrated as it was. The obtained solid was added to dichloromethane and methanol and dissolved, then filtered, and the filtrate was collected. The filtrate was concentrated to obtain the title compound (340 mg, crude product). LC-MS(ESI)[ M + H] + =399.2。

[0128] Step 3: Preparation of tert-butyl 4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-phenylpyrimidin-4-yl)amino)piperidine-1-carboxylate

Chem.

[0129] 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-phenylpyrimidine-4-carboxylic acid (340 mg crude), (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (200 mg, 0.861 mmol, 1 equiv.), EDCI (198 mg, 1.033 mmol, 1.2 equiv.) and HOAt (140 mg, 1.033 mmol, 1.2 equiv.) were dissolved in DMF (2 mL), and the reaction mixture was stirred at 25 °C for 1 hour. The completion of the reaction was monitored by LC-MS. After quenching the reaction mixture by adding water, it was extracted with dichloromethane and methanol. The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the title compound (1 g). LC-MS(ESI)[ M + H] + = 613.4。

[0130] Production Example 7: Preparation of trans-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-((2,4-dimethoxybenzyl)(4-methoxybutyl)amino)-2-phenylpyrimidin-4-yl)methanone (This intermediate can be used, for example, as an intermediate for the compound of Example 45)

[0131] Step 1: Preparation of N-(2,4-dimethoxybenzyl)-4-methoxybutan-1-amine

Chemical Structure

[0132] 4-Chlorobutyl methyl ether (300 mg, 2.45 mmol, 1 equiv.) and 2,4-dimethoxybenzylamine (1.64 g, 9.81 mmol, 4.01 equiv.) were dissolved in acetonitrile (9 ml), potassium carbonate (676 mg, 4.89 mmol, 2 equiv.) and potassium iodide (812 mg, 4.89 mmol, 2 equiv.) were added, and the mixture was stirred at 80 °C overnight (16 h). The formation of the product was monitored by LC-MS. The reaction solution was filtered, and the filtrate was dried by a rotary evaporator. The crude product was separated and purified by flash chromatography (silica gel, MeOH:EA = 1:50) to obtain the title compound (90 mg, yield 11.8%). LC-MS(ESI)[ M + H] + = 254.2。

[0133] Step 2: Preparation of methyl 2-chloro-6-((2,4-dimethoxybenzyl)(4-methoxybutyl)amino)pyrimidine-4-carboxylate

Chemical Structure

[0134] N-(2,4-Dimethoxybenzyl)-4-methoxybutan-1-amine (90 mg, 0.356 mmol, 1 equiv.) and methyl 2,6-dichloropyrimidine-4-carboxylate (220 mg, 1.063 mmol, 3 equiv.) were dissolved in acetonitrile (3 ml), DIPEA (250 mg, 1.934 mmol, 5.4 equiv.) was added, and the mixture was stirred at room temperature (25 °C) for 2 h. After monitoring the completion of the reaction by TLC, the solvent was dried by a rotary evaporator, and the crude product was separated and purified by flash chromatography (silica gel, EA:PE = 1:5) to obtain the title compound (134 mg, yield 75.5%). LC-MS(ESI)[ M + H] + = 424.2。

[0135] Step 3: Preparation of 6-((2,4-dimethoxybenzyl)(4-methoxybutyl)amino)-2-phenylpyrimidine-4-carboxylic acid

Chem.

[0136] In a microwave vial, methyl 2-chloro-6-((2,4-dimethoxybenzyl)(4-methoxybutyl)amino)pyrimidine-4-carboxylate (134 mg, 0.316 mmol, 1 equiv.), benzeneboronic acid (77 mg, 0.632 mmol, 2 equiv.), tetrakis(triphenylphosphine)palladium (36 mg, 0.0312 mmol, 0.1 equiv.) and sodium carbonate (70 mg, 0.660 mmol, 2.09 equiv.) were weighed, capped and purged with nitrogen. Dioxane (1.6 mL) and water (0.4 mL) were added. The reaction was carried out at 120 °C for 1 h under nitrogen using microwave, and the completion of the reaction was monitored by LC-MS. 1 M aqueous hydrochloric acid was added to adjust the pH to 4 - 5. It was filtered and the solvent was dried by rotary evaporator to obtain the crude title compound, which was used directly in the next step. LC-MS (ESI) [M + H] + = 452.2.

[0137] Step 4: Preparation of trans-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-((2,4-dimethoxybenzyl)(4-methoxybutyl)amino)-2-phenylpyrimidin-4-yl)methanone

Chem.

[0138] 6-((2,4-Dimethoxybenzyl)(4-methoxybutyl)amino)-2-phenylpyrimidine-4-carboxylic acid (180 mg, 0.315 mmol, 1 equiv.), EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) (91 mg, 0.475 mmol, 1.51 equiv.) and HOAt (N-hydroxy-7-azabenzotriazole) (65 mg, 0.478 mmol, 1.52 equiv.) were dissolved in DMF (1 mL) and stirred for 5 minutes, then a solution of trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (88 mg, 0.379 mmol, 1.2 equiv.) in DMF (0.6 mL) was added. The reaction mixture was stirred at room temperature for 2 hours. The solvent was dried by rotary evaporator and the crude product was separated and purified by flash chromatography (silica gel, MeOH:DCM = 1:20) to give the title compound (51 mg, yield 24.4%). LC-MS (ESI) [M + H] + = 666.4。

[0139] Production Example 8: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-vinylpyrimidine-4-carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 46).

[0140] Step 1: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-vinylpyrimidine-4-carboxylate

Chemical formula

[0141] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (0.90 g, 2.9 mmol, 1 eq), potassium vinyltrifluoroborate (0.46 g, 3.5 mmol, 1.2 eq), potassium carbonate (0.99 g, 7.2 mmol, 2.5 eq) and [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium (90 mg, 0.1 eq) were dissolved in 1,4-dioxane (30 mL), purged with nitrogen for 2 minutes, heated to 110 °C and reacted for 8 hours. After monitoring the completion of the reaction by LC-MS, it was filtered, and the filter cake was washed twice with a mixed solvent (DCM:MeOH = 10:1) (15 mL). The filtrate was concentrated, and the crude product was first separated on a reverse-phase column with a mixed solvent (DCM:MeOH = 10:1) and concentrated to obtain the title compound (0.85 g, yield 97%). LC-MS(ESI)[ M + H] + = 305.2。

[0142] Step 2: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-vinylpyrimidine-4-carboxylic acid

Chemical formula

[0143] To a mixed solution of tert-butyl 2-vinylpyrimidine-4-carboxylate (0.2 g, 0.66 mmol, 1 eq), acetonitrile (4 mL, 20%) and toluene (16 mL, 80%), tin oxide (0.47 g, 0.79 mmol, 1.2 eq) was added and reacted at 110 °C for 4 hours. After monitoring the completion of the reaction by LC-MS, it was quenched with 2 mL of aqueous KF solution, concentrated, triturated and dried with diethyl ether to obtain the title compound (0.15 g, yield 78%). LC-MS(ESI)[ M + H] + = 291.1。

[0144] Production Example 9: 6-((1-Acetylpiperidin-4-yl)amino)-2-(thiazolo[4,5-c]pyridin-7-yl)pyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 63).

[0145] Step 1: Preparation of 3-Bromo-5-nitropyridine-4-thiol

Chemical formula

[0146] 3-Bromo-4-chloro-5-nitropyridine (10 g, 42.12 mmol, 1.0 equiv.) was dissolved in methanol (80 mL), and sodium hydrosulfide (4.72 g, 84.23 mmol, 2.0 equiv., purity 70%) was added, followed by stirring at 25 °C for 16 hours. After monitoring the completion of the reaction by LC-MS, a sodium hydroxide solution (100 mL, 20%) was added and filtered. The filtrate was acidified to pH = 4 with HCl and filtered, and the cake was dried to obtain the title compound (9 g, yield 90.9%). LC-MS (ESI) [M-H] - = 232.8.

[0147] Step 2: Preparation of 3-Amino-5-bromopyridine-4-thiol

Chemical formula

[0148] 3-Bromo-5-nitropyridine-4-thiol (10 g, 42.5 mmol, 1.0 equiv.) was dissolved in water (60 mL) and HCl (12 M, 25 mL), and stannous chloride (16.13 g, 85.1 mmol, 2.0 equiv.) was added, followed by stirring for 4 hours. After monitoring the completion of the reaction by LC-MS, it was filtered, and the filtrate was concentrated to obtain a crude product (12 g) of the title compound, which was used directly in the next step. LC-MS (ESI) [M-H] - = 202.8.

[0149] Step 3: Preparation of 7-bromothiazolo[4,5-c]pyridine

Chem.

[0150] 3-Amino-5-bromopyridine-4-thiol (10 g, 48.76 mmol, 1.0 equiv.) was dissolved in formic acid (50 mL), and zinc powder (1.59 g, 24.38 mmol, 0.5 equiv.) was added. The reaction was carried out at 100 °C for 1 h. After monitoring the completion of the reaction by LC-MS, it was concentrated, and the crude product was separated and purified by flash chromatography (silica gel, 50% EA in PE) to obtain the title product (2.7 g, yield 25.7%). LC-MS (ESI) [M + H] + = 215.8.

[0151] Step 4: Preparation of thiazolo[4,5-c]pyridin-7-ylboronic acid

Chem.

[0152] 7-Bromothiazolo[4,5-c]pyridine (300 mg, 1.39 mmol, 1.0 equiv.), pinacol diboronic acid (885 mg, 3.49 mmol, 2.5 equiv.), palladium chloride 1,1'-bis(diphenylphosphino)ferrocene (102 mg, 0.14 mmol, 0.1 equiv.) and potassium acetate (274 mg, 2.79 mmol, 2.0 equiv.) were dissolved in dioxane (7 mL) and purged with nitrogen three times. The reaction was carried out at 120 °C for 16 h. After monitoring the completion of the reaction by LC-MS, it was concentrated to obtain 350 mg of the crude title compound, which was used directly in the next step. LC-MS (ESI) [M + H] + = 180.92.

[0153] Step 5: Preparation of Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(thiazolo[4,5-c]pyridin-7-yl)pyrimidine-4-carboxylate

Chem.

[0154] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (300 mg, 0.96 mmol, 1.0 equiv.), thiazolo[4,5-c]pyridin-7-ylboronic acid (259 mg, 1.44 mmol, 1.5 equiv.), 1,1'-bis(diphenylphosphino)ferrocene palladium (76 mg, 0.09 mmol, 0.1 equiv.) and potassium phosphate (1.02 g, 4.80 mmol, 5.0 equiv.) were dissolved in dioxane (6 mL) and water (1.5 mL), and purged with nitrogen three times. The reaction was carried out at 100 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, it was concentrated, and the crude product was separated and purified by flash chromatography (silica gel, 5% MeOH in DCM) to obtain the title compound (390 mg, yield 98.6%). LC-MS (ESI) [M + H] + = 412.85.

[0155] Step 6: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(thiazolo[4,5-c]pyridin-7-yl)pyrimidine-4-carboxylic acid

Chem.

[0156] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(thiazolo[4,5-c]pyridin-7-yl)pyrimidine-4-carboxylate (350 mg, 0.85 mmol, 1.0 equiv.) was dissolved in tetrahydrofuran (5 mL), and an aqueous lithium hydroxide solution (2 M) (24 mg, 1.02 mmol, 1.2 equiv.) was added. The mixture was stirred at 25 °C for 1 hour. After monitoring the completion of the reaction by LC-MS, it was concentrated to obtain 370 mg of the crude title compound, which could be used directly in the next step. LC-MS(ESI)[ M + H] + = 399.0.

[0157] Production Example 10: 4-((1-acetylpiperidin-4-yl)amino)-6-phenyl-1,3,5-triazine-2-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 70)

[0158] First Step: Preparation of 1-(4-((4,6-dichloro-1,3,5-triazin-2-yl)amino)piperidin-1-yl)ethan-1-one

Chemical Structure

[0159] Trispolychlorocyanide (4.0 g, 0.0217 mol, 1 equiv.), 1-(4-aminopiperidin-1-yl)ethan-1-one (3.08 g, 0.0217 mol, 1 equiv.) were added to tetrahydrofuran (110 mL), and N,N-diisopropylethylamine (5.61 g, 0.0434 mol, 2 equiv.) was added. The mixture was stirred at 16 °C for 16 hours, and by TLC, it was detected that the reaction of the starting material was complete. The solvent was dried by a rotary evaporator, and the crude product was purified by column chromatography (DCM:MeOH = 99:1) to obtain the title compound (3.98 g, yield 63.2%). LC-MS(ESI)[ M + H] + = 290.0.

[0160] Step 2: Preparation of 1-(4-((4-chloro-6-phenyl-1,3,5-triazin-2-yl)amino)piperidin-1-yl)ethan-1-one

Chemical formula

[0161] 1-(4-((4,6-dichloro-1,3,5-triazin-2-yl)amino)piperidin-1-yl)ethan-1-one (2 g, 6.893 mmol, 1 equiv.), benzeneboronic acid (0.84 g, 6.893 mmol, 1 equiv.), potassium carbonate (2.86 g, 20.68 mmol, 3 equiv.) were added to 1,4-dioxane / water (45 mL, 4:1), and further, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (252 mg, 0.345 mmol, 0.05 equiv.) was added. The mixture was stirred at 80 °C under nitrogen for 0.75 h, and the completion of the reaction was monitored by TLC. Water was added to the reaction system, and the mixture was extracted three times with ethyl acetate (50 mL). The solvent was dried by a rotary evaporator and purified by column chromatography (DCM:MeOH = 99:1) to obtain the title compound (0.498 g, yield 21.8%). LC-MS(ESI)[M + H] + = 332.0

[0162] Step 3: Preparation of methyl 4-((1-acetylpiperidin-4-yl)amino)-6-phenyl-1,3,5-triazine-2-carboxylate

Chemical formula

[0163] 1-(4-((4-chloro-6-phenyl-1,3,5-triazin-2-yl)amino)piperidin-1-yl)ethan-1-one (0.44 g, 1.33 mmol, 1 equiv.), triethylamine (0.671 g, 6.63 mmol, 5 equiv.) were added to methanol (30 mL), and further, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (97 mg, 0.133 mmol, 0.1 equiv.) was added. The mixture was stirred at carbon monoxide, 100 °C and 2 MPa for 20 h, and the completion of the reaction was monitored by TLC. The solvent was dried by rotary evaporator, and the crude product was purified by column chromatography (DCM:MeOH = 98:2) to obtain the title compound (0.21 g, yield 44.6%). LC-MS(ESI)[ M + H] + = 356.2。

[0164] Step 4: Preparation of 4-((1-acetylpiperidin-4-yl)amino)-6-phenyl-1,3,5-triazine-2-carboxylic acid

Chemical Structure

[0165] Methyl 4-((1-acetylpiperidin-4-yl)amino)-6-phenyl-1,3,5-triazine-2-carboxylate (150 mg, 0.422 mmol, 1 equiv.) was dissolved in tetrahydrofuran / water (5 mL, 4:1), lithium hydroxide (20 mg, 0.844 mmol, 2 equiv.) was added, and the mixture was stirred at 16 °C for 2 h, and the completion of the reaction was monitored by TLC. Water was added to the reaction system, and the mixture was extracted 3 times with ethyl acetate (15 mL). The aqueous phase was adjusted to pH = 4 - 5 with 1M HCl solution and freeze-dried to obtain the crude product of the title compound (0.13 g, yield 90.2%). LC-MS(ESI)[ M + H] + = 342.2。

[0166] Production Example 11: Preparation of 6-((3-(N-Methylacetamido)propyl)amino)-2-phenylpyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 74)

[0167] Step 1: Preparation of tert-Butyl 3-(N-Methylacetamido)propyl)carbamate

Chemical formula

[0168] tert-Butyl (3-(Methylamino)propyl)carbamate (980 mg, 5.3 mmol), DMAP (68 mg, 0.06 mmol) and Et3N (1.07 g, 1.06 mmol) were dissolved in DCM (10 mL). After cooling to 0 °C, acetyl chloride (540 mg, 6.9 mmol) was added dropwise. The reaction mixture was stirred at room temperature (25 - 30 °C) for 2 hours. The reaction was monitored by TLC, and the starting material had disappeared. The mixture was extracted three times with 20 mL of ethyl acetate. The combined ethyl acetate phases were washed once with 10 mL of water and once with 10 mL of saturated brine, dried over anhydrous sodium sulfate for 10 minutes, filtered, concentrated and dried on a rotary evaporator to obtain a crude product, which was then purified by column chromatography to obtain the title compound (1.328 g). 1 1H NMR (400 MHz, CDCl3) δ 5.39 (s, 1H), 3.50 - 3.27 (m, 2H), 3.20 - 3.03 (m, 2H), 3.01 - 2.87 (m, 3H), 2.09 (s, 3H), 1.84 - 1.61 (m, 2H), 1.44 (d, J = 4.6 Hz, 9H).

[0169] Step 2: Preparation of N-(3-Aminopropyl)-N-methylacetamide Trifluoroacetate

Chemical formula

[0170] tert-Butyl (3-(N-methylacetamido)propyl)carbamate (700 mg, 3 mmol) was dissolved in DCM (5 mL), and trifluoroacetic acid (2 mL) was added. The reaction mixture was stirred at room temperature for 3 hours. The reaction was monitored by LC-MS, and the starting material disappeared. The mixture was concentrated and dried on a rotary evaporator to obtain the crude title compound (1.42 g). LC-MS(ESI)[ M + H] + = 131.2。

[0171] Step 3: Preparation of methyl 2-chloro-6-((3-(N-methylacetamido)propyl)amino)pyrimidine-4-carboxylate

Chem.

[0172] Methyl 2,6-dichloropyrimidine-4-carboxylate (300 mg, 1.46 mmol), crude N-(3-aminopropyl)-N-methylacetamidotrifluoroacetate (600 mg, 1.46 mmol) and N,N-diisopropylethylamine (930 mg, 7.3 mmol) were dissolved in acetonitrile (6 mL). The mixture was stirred at 25 °C for 2 hours. It was extracted with ethyl acetate (50 mL × 3), the ethyl acetate phases were combined, washed with water (20 mL), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate for 10 minutes, filtered, and the crude product was separated and purified by column chromatography using DCM:MeOH = 100:2 to obtain the title compound (0.5 g, yield: 85%). LC-MS(ESI)[ M + H] + = 295.2。

[0173] Step 4: Preparation of 6-((3-(N-methylacetamido)propyl)amino)-2-phenylpyrimidine-4-carboxylic acid

Chem.

[0174] Methyl 2-chloro-6-((3-(N-methylacetamido)propyl)amino)pyrimidine-4-carboxylate (500 mg, 1.67 mmol), benzeneboronic acid (244 mg, 2.0 mmol), K2CO3 (460 mg, 3.34 mmol) and Pd(PPh3) 34 (190 mg, 0.167 mmol) were dissolved in 1,4-dioxane (5 mL) and water (1 mL). The mixture was stirred under microwave heating at 120 °C for 1.5 h. The reaction was monitored by LC-MS and the starting materials disappeared. The product was extracted three times with 20 mL portions of ethyl acetate to give an aqueous phase. After concentration and drying on a rotary evaporator, the residue was washed with a mixed solvent of DCM:MeOH = 10:1 (100 mL), and the filtrate was concentrated to obtain the crude title compound (500 mg). LC-MS (ESI) [[ M + H]] + = 343.2.

[0175] Production Example 12: Preparation of 4-amino-1-methylcyclohexan-1-ol (This intermediate can be used, for example, as an intermediate for the compound of Example 86).

[0176] Step 1: Preparation of 8-methyl-1,4-dioxaspiro[4.5]decan-8-ol

Chemical Structure

[0177] At 0 °C, 1,4-dioxaspiro[4.5]decan-8-one (2.0 g, 12.8 mmol, 1 equiv.) was added to anhydrous tetrahydrofuran (40 mL), and then methylmagnesium bromide (3.0 M 2-methyltetrahydrofuran solution) (5.6 mL, 16.64 mmol, 1.3 equiv.) was added. The mixture was stirred at 28 °C for 2 hours. After monitoring the completion of the reaction by TLC, saturated aqueous ammonium chloride solution (15 mL) was added, and the mixture was extracted three times with ethyl acetate (15 mL × 3). The combined organic layers were filtered, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was separated and purified by flash chromatography (silica gel, dichloromethane:methanol (V / V = 10 / 1)) to obtain the title compound (1 g, yield 45%). 1 1H NMR (400 MHz, CDCL3) δ 3.99-3.94 (m, 4H), 1.91-1.87 (m, 2H), 1.73-1.68 (m, 4H), 1.64-1.59 (m, 2H), 1.28 (m, 3H)

[0178] Step 2: Preparation of 4-hydroxy-4-methylcyclohexan-1-one

Chemical Structure

[0179] At 0 °C, 8-methyl-1,4-dioxaspiro[4.5]decan-8-ol (1.0 g, 5.8 mmol, 1.0 equiv.) was added to water (5 mL) and tetrahydrofuran (20.0 mL), 2M HCl (10 mL) was added, and the mixture was stirred at room temperature for 4 hours. After monitoring the completion of the reaction by TLC (dichloromethane:methanol (V / V = 10 / 1)), the reaction mixture was concentrated to 10 mL, washed with saturated sodium bicarbonate solution (30 mL), and extracted three times with ethyl acetate (30 mL × 3). The combined organic phases were filtered and concentrated to obtain the title compound (526 mg, yield: 70%). 11H NMR (400 MHz, CDCl3) δ 2.77 - 2.69 (m, 4H), 2.28 - 2.22 (m, 2H), 2.02 - 1.96 (m, 4H), 1.90 - 1.82 (m, 2H), 1.38 (m, 3H)

[0180] Step 3: Preparation of (S)-N-(4-hydroxy-4-methylcyclohexylene)-2-methylpropane-2-sulfinamide

Chemical formula

[0181] 4-Hydroxy-4-methylcyclohexan-1-one (350 mg, 2.73 mmol, 1.0 equiv) was added to THF (15 mL), then (S)-2-methylpropane-2-sulfinamide (496 mg, 4.1 mmol, 1.5 equiv), Ti(OEt)4 (tetraethyl titanate) (1.56 g, 6.83 mmol, 2.5 equiv) were added, and the mixture was stirred at 80 °C for 3 hours under nitrogen protection. After monitoring the completion of the reaction by LC-MS, water (100 mL) was added, and the mixture was stirred for 15 minutes. Then, it was extracted three times with ethyl acetate (50 mL × 3). The organic phases were combined, filtered, dried over anhydrous sodium sulfate, concentrated to obtain the crude product, and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 1 / 1)) to obtain the title compound (155 mg, yield: 45%). LC-MS (ESI) [[ M + H]] + = 232.1.

[0182] Step 4: Preparation of (S)-N-(4-hydroxy-4-methylcyclohexyl)-2-methylpropane-2-sulfinamide

Chemical formula

[0183] (S)-N-(4-Hydroxy-4-methylcyclohexylene)-2-methylpropane-2-sulfinamide (150 mg, 0.86 mmol, 1.0 equiv) was added to methanol (10 mL), followed by the addition of NaBH4 (66 mg, 1.73 mmol, 2.0 equiv). The mixture was stirred at 28 °C for 10 h. After monitoring the completion of the reaction by LC-MS, the crude product was concentrated and separated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate (V / V = 1 / 1)) to obtain the title compound (155 mg, yield: 45%). LC-MS(ESI)[ M + H] + = 234.2。

[0184] Step 5: Preparation of 4-amino-1-methylcyclohexan-1-ol

Chemical formula

[0185] (S)-N-(4-Hydroxy-4-methylcyclohexyl)-2-methylpropane-2-sulfinamide (200 mg, 0.86 mmol, 1.0 equiv) was added to methanol (10 mL), and then 4M HCl / dioxane (4 mL) was added. The mixture was stirred at 28 °C for 2 h. After monitoring the completion of the reaction by TLC, it was concentrated to obtain the crude product (120 mg) of the title compound.

[0186] Production Example 13: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(cyclopropylethynyl)pyrimidine-4-carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 92)

[0187] Step 1: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(cyclopropylethynyl)pyrimidine-4-carboxylate

Chemical formula

[0188] At 28 °C, under nitrogen protection, methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (180 mg, 1.31 mmol, 1.0 equiv.) was added to DMF (2 mL). Then, ethynylcyclopropane (433 mg, 6.59 mmol, 5.0 equiv.), copper(I) iodide (51 mg, 0.26 mmol, 0.2 equiv.), and tetrakis(triphenylphosphine)palladium(0) (151 mg, 0.131 mmol, 0.1 equiv.) were added, and the mixture was stirred at room temperature for 10 minutes. It was then stirred under microwave heating at 140 °C for 1 hour. After monitoring the completion of the reaction by LC-MS, it was cooled to room temperature, filtered, and the crude product was separated and purified by Prep-HPLC (aqueous C18, 10 mmol / L NH4HCO3, MeCN) to obtain the title compound (170 mg, yield 82.98%). LC-MS (ESI) [M + H] + = 343.3.

[0189] Step 2: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(cyclopropylethynyl)pyrimidine-4-carboxylic acid

Chemical formula

[0190] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(cyclopropylethynyl)pyrimidine-4-carboxylate (170 mg, 0.5 mmol, 1.0 equiv.) was added to water (1 mL), methanol (1.0 mL), and tetrahydrofuran (1.0 mL) at 28 °C. Lithium hydroxide (36 mg, 1.5 mmol, 3.0 equiv.) was added, and the mixture was stirred at room temperature for 2 hours. After monitoring the completion of the reaction by TLC (dichloromethane:methanol (V / V = 10 / 1)), the reaction solution was concentrated to 1 mL, adjusted to pH = 1 - 2 with 1 M hydrochloric acid, and the aqueous phase was lyophilized to obtain the title compound (200 mg, yield: 90%). LC-MS (ESI) [M + H] += 330.2。

[0191] Production Example 14: 6 - ((2 - Acetyl - 2 - azaspiro[3.3]heptan - 6 - yl)amino)-2-(pentan - 3 - yloxy)pyrimidine - 4 - carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 197).

[0192] Step 1: Preparation of 6 - ((2 - (Benzyloxycarbonyl)-2 - azaspiro[3.3]heptan - 6 - yl)amino)-2 - chloropyrimidine - 4 - carboxylic acid

Chemical formula

[0193] Benzyl 6 - ((2 - chloro - 6 - (methoxycarbonyl)pyrimidin - 4 - yl)amino)-2 - azaspiro[3.3]heptane - 2 - carboxylate (220 mg, 0.53 mmol, 1.0 equiv.) and TMSOK (74 mg, 0.58 mmol, 1.1 equiv.) were dissolved in MeCN (8 mL) and reacted at 20 °C for 1 hour. The reaction solution was concentrated to obtain a crude product (240 mg) of the target compound. LC - MS (ESI) [M + H] + = 402.8。

[0194] Step 2: Preparation of 6 - ((2 - azaspiro[3.3]heptan - 6 - yl)amino)-2 - (pentan - 3 - yloxy)pyrimidine - 4 - carboxylic acid

Chemical formula

[0195] 6-((2-(Benzyloxycarbonyl)-2-azaspiro[3.3]heptan-6-yl)amino)-2-chloropyrimidine-4-carboxylic acid (210 mg, 0.52 mmol, 1.0 equiv.) and sodium tert-butoxide (200 mg, 2.09 mmol, 4.0 equiv.) were dissolved in 3-pentanol (10 mL), and reacted at 120 °C for 18 hours under nitrogen protection. The reaction mixture was concentrated, and the crude product was purified by preparative reverse-phase HPLC (10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain the crude target compound (12 mg, yield 7.2%). LC-MS(ESI)[ M + H] + = 321.0。

[0196] Step 3: Preparation of 6-((2-acetyl-2-azaspiro[3.3]heptan-6-yl)amino)-2-(pentan-3-yloxy)pyrimidine-4-carboxylic acid

Chemical formula

[0197] 6-((2-Azaspiro[3.3]heptan-6-yl)amino)-2-(pentan-3-yloxy)pyrimidine-4-carboxylic acid (12 mg, 0.04 mmol, 1.0 equiv.), acetic anhydride (4 mg, 0.04 mmol, 1.1 equiv.) and TEA (6 mg, 0.06 mmol, 1.5 equiv.) were dissolved in DMF (1 mL), and reacted at 20 °C for 0.5 hour. The reaction solution was concentrated to obtain the crude target compound (14 mg). LC-MS(ESI)[ M + H] + = 363.1。

[0198] Production Example 15: Preparation of tert-butyl trans-(1-(6-((1-acetylpiperidin-4-yl)amino)pyrimidine-4-carbonyl)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ylcarbamate (this intermediate can be used, for example, as an intermediate for the compound of Example 151))

[0199] Step 1: Preparation of tert-butyl trans-(4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-yl)carbamate

Chem.

[0200] Benzyl 4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate (trans-3-((tert-butoxycarbonyl)amino)-68 mg, 0.186 mmol, 1 equiv.) was dissolved in methanol (chromatography grade, 5 mL), and Pd / C (10%, 70 mg) was added. The mixture was stirred under hydrogen for 2 hours. The reaction was monitored by TLC. The reaction solution was filtered and dried on a rotary evaporator to obtain a crude product (45 mg) of the title compound, which was used directly in the next step. LC-MS (ESI) [M + H] + = 332.20.

[0201] Step 2: Preparation of tert-butyl trans-(1-(6-((1-acetylpiperidin-4-yl)amino)pyrimidine-4-carbonyl)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-yl)carbamate

Chem.

[0202] 6-((1-Acetylpiperidin-4-yl)amino)pyrimidine-4-carboxylic acid (36 mg, 0.136 mmol, 1 equiv.), EDCI (39 mg, 0.203 mmol, 1.49 equiv.) and HOAt (28 mg, 0.206 mmol, 1.51 equiv.) were dissolved in DMF (2 mL), stirred at room temperature (15 - 20 °C) for 5 minutes, and then a DMF solution (1 mL) of tert-butyl trans-(4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-yl)carbamate (45 mg, 0.174 mmol, 1.7 equiv.) was added. The mixture was stirred at room temperature for 1.5 hours. The solvent was dried using a rotary evaporator, and the crude product was separated and purified by flash chromatography (silica gel, 0 - 7% MeOH in DCM) to obtain tert-butyl trans-(1-(6-((1-acetylpiperidin-4-yl)amino)pyrimidine-4-carbonyl)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-yl)carbamate (53 mg, yield 67.3%).

[0203] Production Example 16: 2-(1-Acetylpiperidin-4-yl)-1H-benzo[d]imidazole-6-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 155).

[0204] Step 1: Preparation of methyl 4-(1-acetylpiperidine-4-carboxamido)-3-aminobenzoate

Chemical Structure

[0205] Methyl 3,4-diaminobenzoate (200 mg, 1.20 mmol, 1 equiv.), 1-acetyl-4-piperidinecarboxylic acid (206 mg, 1.20 mmol, 1.0 equiv.), EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) (277 mg, 1.44 mmol, 1.2 equiv.) and HOAt (N-hydroxy-7-azabenzotriazole) (196 mg, 1.44 mmol, 1.2 equiv.) were dissolved in DCM (6 mL) and reacted at 25 °C for 16 h. The precipitate was collected by filtration and the filter cake was dried to obtain the crude title compound (300 mg). LC-MS (ESI) [M + H] + = 320.2.

[0206] Step 2: Preparation of methyl 2-(1-acetylpiperidin-4-yl)-1H-benzo[d]imidazole-6-carboxylate

Chem.

[0207] Methyl 4-(1-acetylpiperidine-4-carboxamido)-3-aminobenzoate (250 mg, 0.78 mmol, 1 equiv.) was dissolved in acetic acid (6 mL) and reacted at 65 °C for 5 h. The reaction mixture was neutralized to pH = 6 - 7 with saturated aqueous NaHCO3 and extracted with dichloromethane (3 * 10 mL). The combined organic phases were washed with brine, dried over Na2SO4 and concentrated to obtain the title compound (170 mg, yield 72.1%). LC-MS (ESI) [M + H] + = 302.2.

[0208] Step 3: Preparation of 2-(1-acetylpiperidin-4-yl)-1H-benzo[d]imidazole-6-carboxylic acid

Chem.

[0209] Methyl 2-(1-acetylpiperidin-4-yl)-1H-benzo[d]imidazole-6-carboxylate (80 mg, 0.27 mmol, 1.0 equiv.) was dissolved in tetrahydrofuran (1 mL), and an aqueous lithium hydroxide solution (2 M) (13 mg, 0.53 mmol, 2.0 equiv.) was added. The mixture was stirred at 60 °C for 1 hour. After monitoring the completion of the reaction by LC-MS, it was concentrated to obtain the crude product (90 mg) of the title compound. LC-MS (ESI) [M + H] + = 288.2.

[0210] Production Example 17: Preparation of 6-((1-(tetrahydro-2H-pyran-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 157)

[0211] First Step: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylate

Chemical Structure

[0212] Methyl 6-chloropyrimidine-4-carboxylate (4 g, 23.17 mmol, 1 equiv.) and tert-butyl 4-aminopiperidine-1-carboxylate (4.9 g, 24.33 mmol, 1.05 equiv.) were dissolved in acetonitrile (40 ml), and DIPEA (8.99 g, 69.51 mmol, 3 equiv.) was added. The reaction mixture was stirred at 80 °C for 3 hours. The completion of the reaction was monitored by LC-MS. Water was added to the reaction solution to quench it, and it was extracted with ethyl acetate. The organic layers were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was separated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate = 1:5) to obtain the title compound (5.56 g, yield 71%). LC-MS (ESI) [M + H] += 337.2。

[0213] Step 2: Preparation of Methyl 6-(Piperidin-4-ylamino)pyrimidine-4-carboxylate

Chem.

[0214] Methyl 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylate (5.46 g, 16.2 mmol, 1 equiv.) was dissolved in dichloromethane (40 mL), and trifluoroacetic acid (10 mL) was added. The reaction mixture was stirred at 20 °C for 2 hours. The completion of the reaction was monitored by LC-MS. Water was added to the reaction solution, and the pH was adjusted to 8 - 9 with saturated sodium hydrogen carbonate solution, followed by extraction with ethyl acetate. After concentrating the aqueous phase, the crude product (11 g) of the title compound was obtained. LC-MS (ESI) [M + H] + = 237.1。

[0215] Step 3: Preparation of Methyl 6-((1-(Tetrahydro-2H-pyran-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylate

Chem.

[0216] Methyl 6-(piperidin-4-ylamino)pyrimidine-4-carboxylate (923 mg, crude) was dissolved in dichloromethane (5 mL), then triethylamine (195 mg, 1.907 mmol, 1.5 equiv.) was added, and the reaction was cooled to around -10 °C. Then, while maintaining the temperature of the reaction system not exceeding 0 °C, tetrahydro-4H-pyran-4-carbonyl chloride (190 mg, 1.271 mmol, 1 equiv.) was slowly added to the reaction solution. The reaction was gradually warmed to room temperature (20 °C) and stirred for 1 hour. After monitoring the completion of the reaction by LC-MS, water was added to the reaction solution to quench it, then extracted with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, then filtered and concentrated to obtain the crude title compound (340 mg). LC-MS(ESI)[ M + H] + = 349.2。

[0217] Step 4: Preparation of 6-((1-(tetrahydro-2H-pyran-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid

Chemical Structure

[0218] Methyl 6-((1-(tetrahydro-2H-pyran-4-carbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylate (340 mg, crude) was dissolved in dichloromethane (15 mL) and methanol (3 mL), and TMSOK (potassium trimethylsilanolate) (340 mg, 1.172 mmol, 1.2 equiv.) was added. The mixture was stirred at room temperature (20 °C) for 1 hour. The completion of the reaction was monitored by TLC. The reaction solution was concentrated to obtain a solid, which was dissolved by adding water. Then, 1N hydrochloric acid was added to adjust the pH to 5 - 6, and after extraction with dichloromethane, the solid obtained by concentrating the aqueous phase was added to dichloromethane and methanol to dissolve it. The solid impurities were removed by filtration, and the filtrate was concentrated to obtain the crude title compound (300 mg), which was used directly in the next step. LC-MS (ESI) [M + H] + = 335.2。

[0219] Production Example 18: Preparation of 6 - ((1 - (thiazole - 5 - carbonyl) piperidin - 4 - yl) amino) pyrimidine - 4 - carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 167)

[0220] Step 1: Preparation of methyl 6 - ((1 - (thiazole - 5 - carbonyl) piperidin - 4 - yl) amino) pyrimidine - 4 - carboxylate

Chemical Structure

[0221] Methyl 6 - (piperidin - 4 - ylamino) pyrimidine - 4 - carboxylate (100 mg, 0.42 mmol, 1.0 equiv.), thiazole - 5 - carboxylic acid (67 mg, 0.42 mmol, 1.0 equiv.), EDCI (1 - (3 - dimethylaminopropyl) - 3 - ethylcarbodiimide hydrochloride) (60 mg, 0.32 mmol, 1.5 equiv.) and HOAt (N - hydroxy - 7 - azabenzotriazole) (43 mg, 0.32 mmol, 1.5 equiv.) were dissolved in DMF (N,N - dimethylformamide) (1 mL) and reacted at 25 °C for 1 hour. After monitoring the completion of the reaction by LC - MS, it was concentrated, and the crude product was separated and purified by flash chromatography (silica gel, 5% MeOH in DCM) to obtain the title product (90 mg, yield 61.2%). LC-MS (ESI) [M + H] + = 348.2。

[0222] Step 2: Preparation of 6 - ((1 - (thiazole - 5 - carbonyl) piperidin - 4 - yl) amino) pyrimidine - 4 - carboxylic acid

Chemical Structure

[0223] Methyl 6 - ((1 - (thiazole - 5 - carbonyl)piperidin - 4 - yl)amino)pyrimidine - 4 - carboxylate (90 mg, 0.26 mmol, 1.0 equiv.) was dissolved in tetrahydrofuran (1 mL), and an aqueous lithium hydroxide solution (2 M) (12 mg, 0.52 mmol, 2.0 equiv.) was added, followed by stirring at 25 °C for 1 hour. After monitoring the completion of the reaction by LC - MS, it was concentrated to obtain the crude title compound (90 mg). LC - MS(ESI)[M + H] + = 334.0。

[0224] Production Example 19: Preparation of 6 - ((1 - acetylpiperidin - 4 - yl)amino)-2 - (cyclopentylthio)pyrimidine - 4 - carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 198)

Chemical Structure

[0225] Methyl 6 - ((1 - acetylpiperidin - 4 - yl)amino)-2 - chloropyrimidine - 4 - carboxylate (100 mg, 0.32 mmol, 1.0 equiv.), cyclopentanethiol (163 mg, 1.60 mmol, 5.0 equiv.) and Cs2CO3 (208 mg, 0.64 mmol, 2.0 equiv.) were dissolved in EtOH (0.7 mL) and reacted at 70 °C for 16 hours. The reaction solution was suction - filtered, dilute hydrochloric acid was added to the filtrate to adjust it to acidic, and then concentrated to obtain the crude target compound (340 mg). LC - MS(ESI)[M + H] + = 365.2。

[0226] Production Example 20: 6 - ((2 - acetyl - 2 - azaspiro[3.3]heptan - 6 - yl)amino)-2 - isopropoxypyrimidine - 4 - carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 200)

[0227] Step 1: Preparation of 2-Isopropoxy-6-((2-(isopropoxycarbonyl)-2-azaspiro[3.3]heptan-6-yl)amino)pyrimidine-4-carboxylic acid

Chem.

[0228] Benzyl 6-((2-chloro-6-(methoxycarbonyl)pyrimidin-4-yl)amino)-2-azaspiro[3.3]heptane-2-carboxylate (150 mg, 0.36 mmol, 1.0 equiv.) and sodium tert-butoxide (138 mg, 1.44 mmol, 4.0 equiv.) were dissolved in i-PrOH (15 mL) and reacted at 100 °C for 3 hours under nitrogen protection. Dilute hydrochloric acid was used to adjust the reaction solution to acidity and concentrated to obtain 270 mg of the crude product of the target compound. LC-MS (ESI) [M + H] + = 379.2.

[0229] Step 2: Preparation of Isopropyl 6-((6-(3R,4R-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-isopropoxypyrimidin-4-yl)amino)-2-azaspiro[3.3]heptane-2-carboxylate

Chem.

[0230] 2-Isopropoxy-6-((2-(isopropoxycarbonyl)-2-azaspiro[3.3]heptan-6-yl)amino)pyrimidine-4-carboxylic acid (250 mg, 0.33 mmol, 1.0 equiv.), (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (77 mg, 0.33 mmol, 1.0 equiv.), EDCI (95 mg, 0.50 mmol, 1.5 equiv.) and HOAt (67 mg, 0.50 mmol, 1.5 equiv.) were dissolved in DMF (3.7 mL) and reacted at 20° C. for 1 h. The reaction was quenched with saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, the organic phase was concentrated, and the crude product was separated and purified by flash chromatography (silica gel, DCM:MeOH = 30:1) to obtain the title compound (70 mg, yield 35.8%). LC-MS(ESI)[M+H] + = 593.3.

[0231] Third step: Preparation of 6-((2-azaspiro[3.3]heptan-6-yl)amino)-2-isopropoxypyrimidine-4-carboxylic acid [ka]

[0232] Isopropyl 6-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-isopropoxypyrimidin-4-yl)amino)-2-azaspiro[3.3]heptane-2-carboxylate (55 mg, 0.09 mmol, 1.0 equiv.) was dissolved in MeOH (3 mL) and aqueous potassium hydroxide (2M) (3 mL) and reacted at 80° C. for 20 hours. The reaction solution was acidified and suction filtered, and the filtrate was concentrated to obtain 170 mg of a crude product of the target compound. LC-MS(ESI)[M+H] + = 293.1.

[0233] Step 4: Preparation of 6-((2-acetyl-2-azaspiro[3.3]heptan-6-yl)amino)-2-isopropoxypyrimidine-4-carboxylic acid

Chem.

[0234] 6-((2-azaspiro[3.3]heptan-6-yl)amino)-2-isopropoxypyrimidine-4-carboxylic acid (140 mg, 0.09 mmol, 1.0 equiv.), acetic anhydride (10 mg, 0.09 mmol, 1.0 equiv.) and TEA (15 mg, 0.14 mmol, 1.5 equiv.) were dissolved in MeOH (5 mL) and reacted at 20 °C for 0.5 h. The reaction solution was concentrated to obtain the crude product (32 mg) of the target compound. LC-MS (ESI) [M + H] + = 335.2.

[0235] Production Example 21: 2-isopropoxy-6-((1-(pyrimidin-4-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 204)

[0236] Step 1: Preparation of 2-isopropoxy-6-((1-(pyrimidin-4-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid

Chem.

[0237] A solution of methyl 2-chloro-6-((1-(pyrimidin-4-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylate (0.20 g, 0.57 mmol, 1 equiv.) in isopropanol (8 ml, 100%) was added with sodium tert-butoxide (0.33 g, 3.44 mmol, 6 equiv.), and nitrogen was replaced twice, followed by reacting at 100 °C for 20 hours. After monitoring the completion of the reaction by LC-MS, it was concentrated, and the crude product was adjusted to pH = 5 with 1 M hydrochloric acid and suction dried to obtain the title compound (0.1 g, yield 48.7%). LC-MS(ESI)[ M + H] + = 359.1。

[0238] Production Example 22: 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-methoxypyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 220).

[0239] Step 1: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate

Chemical formula

[0240] Methyl 2,6-dichloropyrimidine-4-carboxylate (1.086 g, 5.24 mmol, 1.05 equiv.) and tert-butyl 4-aminopiperidine-1-carboxylate (1 g, 4.99 mmol, 1.05 equiv.) were dissolved in acetonitrile (40 mL), and DIPEA (1.94 g, 14.97 mmol, 3 equiv.) was added. The reaction mixture was stirred at 80 °C for 3 h. The completion of the reaction was monitored by LC-MS. Water was added to the reaction solution to quench it, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was separated and purified by flash chromatography (silica gel, petroleum ether:ethyl acetate = 0 - 20%) to give the title compound (1.5 g, 80% yield). LC-MS (ESI) [M + H] + = 371.0。

[0241] Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-methoxypyrimidine-4-carboxylate

Chemical Structure

[0242] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (500 mg, 1.348 mmol, 1.0 equiv.) and sodium methoxide (110 mg, 2.022 mmol, 2 equiv.) were dissolved in methanol (10 mL), and the reaction mixture was stirred at 80 °C for 16 h. The completion of the reaction was monitored by LC-MS. Water was added to the reaction solution to quench it, and the mixture was extracted with methylene chloride. After combining the organic phases, the mixture was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product (400 mg) of the title compound. LC-MS (ESI) [M + H] + = 367.2。

[0243] Step 3: Preparation of 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-methoxypyrimidine-4-carboxylic Acid

Chem.

[0244] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-methoxypyrimidine-4-carboxylate (400 mg) was dissolved in acetonitrile (20 mL), and TMSOK (potassium trimethylsilanolate) (169 mg, 1.307 mmol, 1.2 equiv.) was added. The mixture was stirred at room temperature (20 °C) for 1 hour. The completion of the reaction was monitored by TLC. The reaction solution was concentrated to obtain a solid, which was dissolved by adding water. Then, 1N hydrochloric acid was added to adjust the pH to 5 - 6, and the mixture was extracted with dichloromethane. After concentrating the aqueous phase, dichloromethane and methanol were added to the obtained solid to dissolve it, and solid impurities were removed by filtration. The filtrate was concentrated to obtain the crude title compound (350 mg), which was used directly in the next step. LC-MS (ESI) [M + H] + = 353.2.

[0245] Production Example 23: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethoxy)pyrimidine-4-carboxylic Acid (This intermediate can be used, for example, as an intermediate for the compound of Example 222)

[0246] Step 1: Preparation of Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-hydroxypyrimidine-4-carboxylate

Chem.

[0247] Methyl 6-[(1-acetylpiperidin-4-yl)amino]-2-chloropyrimidine-4-carboxylate (1 g, 3.2 mmol, 1 equiv.) was dissolved in formic acid (10 mL, 100%) and heated at 100 °C for 12 h. After monitoring the completion of the reaction by LC-MS, it was concentrated, and the crude product was treated with (C18, 5 mmol / L NH3 in aqueous solution / acetonitrile) to obtain the title compound (200 mg, 21.3%). LC-MS(ESI)[ M + H] + = 295.1。

[0248] Step 2: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethoxy)pyrimidine-4-carboxylate

Chemical formula

[0249] 3,3-Dimethyl-1-(trifluoromethyl)-1,2-benziodoxolane (0.22 g, 0.68 mmol, 1 equiv.) was added to a solution of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-hydroxypyrimidine-4-carboxylate (200.13 mg, 0.68 mmol, 1 equiv.) in nitromethane (3 mL, 100%) at room temperature. After purging with nitrogen, it was heated at 100 °C for 12 h, 3,3-dimethyl-1-(trifluoromethyl)-1,2-benziodoxolane (0.011 g, 0.034 mmol, 0.5 equiv.) was added, and it was heated at 100 °C for 12 h to continue the reaction. After monitoring the completion of the reaction by LC-MS, it was concentrated and dried on a rotary evaporator, and purified by column chromatography using 10% DCM / MeOH to obtain methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethoxy)pyrimidine-4-carboxylate (35 mg, 14.2%). LC-MS(ESI)[ M + H] + = 363.1。

[0250] Step 3: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethoxy)pyrimidine-4-carboxylic acid

Chem.

[0251] tert-Butyl 2-(trifluoromethoxy)pyrimidine-4-carboxylate (0.069 g, 0.12 mmol, 1.2 equiv.) was added to a mixed solution of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethoxy)pyrimidine-4-carboxylate (25 mg, 0.07 mmol, 1 equiv.), acetonitrile (1 mL, 20%) and toluene (4 mL, 80%). The reaction was carried out at 110 °C for 12 hours. After monitoring the completion of the reaction by LC-MS, the crude product was quenched with 1 mL of aqueous potassium fluoride solution, concentrated, the crude product was pulped with diethyl ether, extracted and dried to obtain 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethoxy)pyrimidine-4-carboxylic acid (25 mg, yield 74.3%). LC-MS (ESI) [M + H] + = 349.0。

[0252] Production Example 24: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-ethoxypyrimidine-4-carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 235)

Chem.

[0253] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (100 mg, 0.06 mmol, 1.0 equiv.) was dissolved in ethanol (3 mL) at 20 °C, sodium ethoxide was added, and the mixture was stirred at 80 °C for 10 h. After monitoring the completion of the reaction by LC-MS, the product was concentrated, and the crude product was separated and purified by Prep-HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile), and freeze-dried to obtain the title compound (60 mg, yield: 61%). LC-MS (ESI) [M + H] + = 309.3.

[0254] Production Example 25: Preparation of 2-(pentan-3-yloxy)-6-(((1-(thiazol-2-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 251)

[0255] Step 1: Preparation of tert-butyl (1-(thiazol-2-yl)piperidin-4-yl)carbamate

Chemical formula

[0256] tert-Butyl piperidin-4-ylcarbamate (1 g, 4.993 mmol, 1.0 equiv.) was dissolved in DMF (10 mL), 2-bromothiazole (0.82 g, 4.993 mmol, 1.0 equiv.) and potassium carbonate (3.45 g, 24.997 mmol, 5.0 equiv.) were added. The mixture was stirred at 120 °C for 16 h. After monitoring the completion of the reaction by LC-MS, it was concentrated and purified using a chromatography column (MeOH:DCM = 50%) to obtain the title compound (670 mg, yield 47.4%). LC-MS (ESI) [M + H] + = 284.0.

[0257] Step 2: Preparation of 1-(thiazol-2-yl)piperidin-4-amine

Chem.

[0258] tert-Butyl (1-(thiazol-2-yl)piperidin-4-yl)carbamate (660 mg, 2.329 mmol, 1.0 equiv.) was dissolved in dioxane (10 mL), and a dioxane solution of hydrochloric acid (4 mL) was added, followed by stirring at 25 °C for 2 h. After monitoring the completion of the reaction by LC-MS, it was concentrated to obtain the crude title compound (569 mg). LC-MS (ESI) [M + H] + = 183.9.

[0259] Step 3: Preparation of methyl 2-chloro-6-((1-(thiazol-2-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylate

Chem.

[0260] Under the condition of 0 °C, methyl 2,6-dichloropyrimidine-4-carboxylate (632 mg, 3.056 mmol, 1.0 equiv.) was added to acetonitrile (10 mL), 1-(thiazol-2-yl)piperidin-4-amine (560 mg, 3.056 mmol, 1.0 equiv.) and DIPEA (0.39 g, 6.111 mmol, 2.0 equiv.) were added, and the mixture was stirred at 0 °C for 16 h. After monitoring the completion of the reaction by LC-MS, it was concentrated to obtain the title compound (150 mg, yield: 13.9%). LC-MS (ESI) [M + H] + = 353.9.

[0261] Step 4: Preparation of 2-(pentan-3-yloxy)-6-(((1-(thiazol-2-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid [Chemical formula]

[0262] Methyl 2-chloro-6-((1-(thiazol-2-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylate (135 mg, 0.381 mmol, 1.0 equiv.) was dissolved in 3-pentanol (2 mL), and sodium tert-butoxide (220 mg, 2.289 mmol, 6.0 equiv.) was added. The mixture was stirred at 110 °C for 1 hour. After monitoring the completion of the reaction by LC-MS, it was concentrated to obtain the title compound (50 mg, yield 33.5%). LC-MS (ESI) [M + H] + = 392.2.

[0263] Production Example 26: 2-((1-acetylpiperidin-4-yl)amino)-6-(cyclopentyloxy)isonicotinic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 283)

[0264] Step 1: Preparation of 2,6-dichloroisonicotinic acid [Chemical formula]

[0265] Methyl 2,6-dichloroisonicotinate (1.5 g, 7.28 mmol, 1.0 equiv.) was dissolved in tetrahydrofuran (10 mL), and an aqueous lithium hydroxide solution (2 M) (209 mg, 8.73 mmol, 1.2 equiv.) was added. The mixture was stirred at 25 °C for 1 hour. After monitoring the completion of the reaction by LC-MS, it was concentrated to obtain the crude title compound (1.7 g). LC-MS (ESI) [M + H] + = 191.9.

[0266] Step 2: Preparation of 2-chloro-6-(cyclopentyloxy)isonicotinic acid [Chemical formula]

[0267] Sodium hydride (1.5 g, 37.5 mmol, 4.0 equiv., purity 60%) was dissolved in dioxane (40 mL), and cyclopentanol (1.21 g, 14.1 mmol, 1.5 equiv.) was added dropwise at room temperature and stirred for 15 minutes. Then, 2,6-dichloroisonicotinic acid (1.8 g, 9.38 mmol, 1.0 equiv.) was added and the mixture was purged with nitrogen three times. The reaction was carried out at 80 °C for 16 hours. After monitoring the completion of the reaction by LC-MS, it was diluted with ethyl acetate (50 mL), washed once with 1 M hydrochloric acid and saturated brine (50 mL), the organic layer was dried over anhydrous sodium sulfate and concentrated to obtain the crude product (2.5 g) of the title compound. LC-MS (ESI) [M + H] + = 242.0.

[0268] Step 3: Preparation of methyl 2-chloro-6-(cyclopentyloxy)isonicotinate

Chemical formula

[0269] 2-Chloro-6-(cyclopentyloxy)isonicotinic acid (2.3 g, 9.52 mmol, 1.0 equiv.), methyl iodide (1.62 g, 11.42 mmol, 1.2 equiv.) and potassium carbonate (5.26 g, 38.07 mmol, 4.0 equiv.) were dissolved in DMF (40 mL). The reaction was carried out at 25 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, it was concentrated and the crude product was purified by flash chromatography (silica gel, 5% EA in PE) to obtain the title compound (2 g, yield 84.9%). LC-MS (ESI) [M + H] + = 256.0.

[0270] Step 4: Preparation of methyl 2-((1-acetylpiperidin-4-yl)amino)-6-(cyclopentyloxy)isonicotinate [Chemical formula]

[0271] Methyl 2-chloro-6-(cyclopentyloxy)isonicotinate (1.4 g, 5.48 mmol, 1.0 equiv.), 1-acetylpiperidin-4-amine (1.17 g, 8.21 mmol, 1.5 equiv.), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (1.37 g, 2.19 mmol, 0.4 equiv.), palladium(II) acetate (246 mg, 1.10 mmol, 0.2 equiv.) and cesium carbonate (5.38 g, 16.42 mmol, 3.0 equiv.) were dissolved in dioxane (30 mL) and purged with nitrogen three times. The reaction was carried out at 100 °C for 4 h. After monitoring the completion of the reaction by LC-MS, the reaction mixture was concentrated and the crude product was separated and purified by reverse-phase HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to give the title compound (170 mg, yield 8.6%). LC-MS (ESI) [M + H] + = 362.0.

[0272] Step 5: Preparation of 2-((1-acetylpiperidin-4-yl)amino)-6-(cyclopentyloxy)isonicotinic acid [Chemical formula]

[0273] Methyl 2-((1-acetylpiperidin-4-yl)amino)-6-(cyclopentyloxy)isonicotinate (150 mg, 0.42 mmol, 1.0 equiv.) was dissolved in tetrahydrofuran (1 mL), and an aqueous lithium hydroxide solution (2 M) (12 mg, 0.50 mmol, 1.2 equiv.) was added. The mixture was stirred at 25 °C for 1 h. After monitoring the completion of the reaction by LC-MS, the reaction mixture was concentrated to give the crude title compound (150 mg). LC-MS (ESI) [M + H] + = 348.0.

[0274] Production Example 27: Preparation of ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-isopropoxy-6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone (This intermediate can be used, for example, as an intermediate for the compound of Example 293)

[0275] Step 1: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate

Chemical formula

[0276] tert-Butyl 4-aminopiperidine-1-carboxylate (873.79 mg, 4.37 mmol, 0.9 equiv.) and N,N-diisopropylethylamine (1.88 g, 14.55 mmol, 3.0 equiv.) were added and stirred at 25 °C for 2 hours. After monitoring the completion of the reaction by TLC and LC-MS, the solvent was centrifuged and separated and purified by Prep-TLC (EA:PE = 20%) to obtain the title compound (1.5 g, yield 83.7%). LC-MS (ESI) [M + H] + = 371.3.

[0277] Step 2: Preparation of 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylic acid

Chemical formula

[0278] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (500 mg, 1.347 mmol, 1.0 equiv.) was dissolved in acetonitrile (5 mL), potassium trimethylsilyl (0.1729 g, 1.347 mmol, 1.0 equiv.) was added, and the mixture was stirred at 25 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, it was concentrated, dissolved in a 10% dichloromethane-methanol solution, filtered, and the filtrate was concentrated to obtain the crude title compound (480 mg, yield 99.8%). LC-MS (ESI) [M + H] + = 357.2。

[0279] Step 3: Preparation of 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-isopropoxypyrimidine-4-carboxylic acid

Chemical Structure

[0280] 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylic acid (480 mg, 1.35 mmol, 1.0 equiv.) was dissolved in isopropanol (5 mL), potassium tert-butoxide (628.8 mg, 5.4 mmol, 4.0 equiv.) was added, and the mixture was stirred at 90 °C for 16 hours. After monitoring the completion of the reaction by LC-MS, it was concentrated and separated and purified using Prep-TLC (MeOH:DCM = 10%) to obtain the title compound (529 mg, yield 99.3%). LC-MS (ESI) [M + H] + = 381.3。

[0281] Step 4: Preparation of tert-butyl 4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-isopropoxypyrimidin-4-yl)amino)piperidine-1-carboxylate [Chemical formula]

[0282] 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-isopropoxypyrimidine-4-carboxylic acid (200 mg, 0.526 mmol, 1.0 equiv.) was dissolved in N,N-dimethylformamide (2 mL), and 3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (146.6 mg, 0.631 mmol, 1.2 equiv.), HATU (299.8 mg, 0.789 mmol, 1.5 equiv.) and N,N-diisopropylethylamine (203.8 mg, 1.577 mmol, 3.0 equiv.) were added, and the mixture was stirred at room temperature for 2 hours. After monitoring the completion of the reaction by LC-MS, the mixture was extracted and concentrated, and purified by reverse phase to obtain the title compound (80 mg, yield 25.6%). LC-MS (ESI) [M + H] + = 595.4.

[0283] Step 5: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-isopropoxy-6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone [Chemical formula]

[0284] tert-Butyl 4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-isopropoxypyrimidin-4-yl)amino)piperidine-1-carboxylate (80 mg, 0.134 mmol, 1.0 equiv.) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (0.5 mL) was added, and the mixture was stirred at 25 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, the mixture was concentrated to obtain the crude product (70 mg) of the title compound. LC-MS (ESI) [M + H] + = 495.3。

[0285] Production Example 28: Preparation of 6-((1-(Cyclobutanecarbonyl)piperidin-4-yl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 304)

[0286] Step 1: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate

Chemical formula

[0287] Methyl 2,6-dichloropyrimidine-4-carboxylate (1000 mg, 4.85 mmol, 1.0 equiv.) was dissolved in acetonitrile (10 mL), and tert-butyl 4-aminopiperidine-1-carboxylate (873.79 mg, 4.37 mmol, 0.9 equiv.) and N,N-diisopropylethylamine (1876.95 mg, 14.55 mmol, 3.0 equiv.) were added, followed by stirring at 25 °C for 2 hours. After monitoring the completion of the reaction by TLC and LC-MS, the solvent was centrifuged and separated and purified by Prep-TLC (EA:PE = 20%) to obtain the title compound (1500 mg, yield 83.7%). LC-MS (ESI) [M + H] + = 371.3。

[0288] Step 2: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylate

Chemical formula

[0289] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (500 mg, 1.35 mmol, 1.0 equiv.) was dissolved in acetonitrile (5 mL), pyrrolidine (105.72 mg, 1.49 mmol, 1.1 equiv.) and N,N-diisopropylethylamine (348.3 mg, 2.7 mmol, 2.0 equiv.) were added, and the mixture was stirred at 90 °C for 16 h. After monitoring the completion of the reaction by LC-MS, it was concentrated and separated by Prep-TLC (EA:PE = 30%) and purified to obtain the title compound (373 mg, yield 68.2%). LC-MS(ESI)[ M + H] + = 406.4。

[0290] Step 3: Preparation of methyl 6-(piperidin-4-ylamino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylate

Chemical Structure

[0291] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylate (50 mg, 0.123 mmol, 1.0 equiv.) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (0.25 mL) was added, and the mixture was stirred at 25 °C for 2 h. After monitoring the completion of the reaction by LC-MS, it was extracted and concentrated to obtain the crude title compound (50 mg, yield 100%). LC-MS(ESI)[ M + H] + = 306.4。

[0292] Step 4: Preparation of methyl 6-((1-(cyclobutanecarbonyl)piperidin-4-yl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylate

Chemical Structure

[0293] Methyl 6-(piperidin-4-ylamino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylate (50 mg, 0.164 mmol, 1.0 equiv.) was dissolved in dichloromethane (1 mL), N,N-diisopropylethylamine (63.44 mg, 0.49 mmol, 3.0 equiv.) was added, and the mixture was stirred at 0 °C for 15 minutes. Under ice bath conditions, cyclobutanecarbonyl chloride (21.29 mg, 0.18 mmol, 1.1 equiv.) was added dropwise and the mixture was stirred for 1 hour. After monitoring the completion of the reaction by LC-MS, the mixture was extracted and concentrated to obtain the crude product of the title compound (62 mg, yield: 97.79%). LC-MS (ESI) [M + H] + = 388.3.

[0294] Step 5: Preparation of 6-((1-(cyclobutanecarbonyl)piperidin-4-yl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylic acid

Chemical Structure

[0295] Methyl 6-((1-(cyclobutanecarbonyl)piperidin-4-yl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylate (62 mg, 0.16 mmol, 1.0 equiv.) was dissolved in acetonitrile (1 mL), potassium trimethylsilanolate (24.8 mg, 0.24 mmol, 1.5 equiv.) was added, and the mixture was stirred at 25 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, the mixture was concentrated and the pH was adjusted to weakly acidic with dilute hydrochloric acid solution to obtain the crude product of the title compound (62 mg). LC-MS (ESI) [M + H] + = 374.3.

[0296] Production Example 29: 2-(Piperidin-1-yl)-6-((1-(pyrimidin-4-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 318).

[0297] Step 1: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate

Chemical formula

[0298] Methyl 2,6-dichloropyrimidine-4-carboxylate (1 g, 4.85 mmol, 1.0 equiv.) was dissolved in acetonitrile (10 mL), and tert-butyl 4-aminopiperidine-1-carboxylate (873.79 mg, 4.37 mmol, 0.9 equiv.) and N,N-diisopropylethylamine (1.88 g, 14.55 mmol, 3.0 equiv.) were added, and the mixture was stirred at 25 °C for 2 hours. After monitoring the completion of the reaction by TLC and LC-MS, the solvent was centrifuged and purified by flash silica gel column (EA:PE = 1:5) to obtain the title compound (1.5 g, yield 83.7%). LC-MS (ESI) [M + H] + = 371.3.

[0299] Step 2: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-(piperidin-1-yl)pyrimidine-4-carboxylate

Chemical formula

[0300] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (500 mg, 1.35 mmol, 1.0 equiv.) was dissolved in acetonitrile (5 mL), hexahydropyridine (126.3 mg, 1.48 mmol, 1.1 equiv) and N,N-diisopropylethylamine (348.3 mg, 2.7 mmol, 2.0 equiv.) were added, and the mixture was stirred at 90 °C for 16 h. After monitoring the completion of the reaction by LC-MS, it was concentrated and separated and purified using a flash silica gel column (EA:PE = 1:3) to obtain the title compound (300 mg, yield 53%). LC-MS(ESI)[ M + H] + = 420.2。

[0301] Step 3: Preparation of methyl 2-(piperidin-1-yl)-6-(piperidin-4-ylamino)pyrimidine-4-carboxylate

Chemical formula

[0302] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-(piperidin-1-yl)pyrimidine-4-carboxylate (150 mg, 0.358 mmol, 1.0 equiv.) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (0.25 mL) was added, and the mixture was stirred at 25 °C for 2 h. After monitoring the completion of the reaction by LC-MS, water was added and the mixture was extracted with dichloromethane and concentrated to obtain the crude product (140 mg) of the title compound. LC-MS(ESI)[ M + H] + = 320.2。

[0303] Step 4: Preparation of methyl 2-(piperidin-1-yl)-6-((1-(pyrimidin-4-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylate

Chemical formula

[0304] Methyl 2-(piperidin-1-yl)-6-(piperidin-4-ylamino)pyrimidine-4-carboxylate (100 mg, 0.31 mmol, 1.0 equiv.) was dissolved in dichloromethane (1 mL), DIPEA (121.40 mg, 0.94 mmol, 3.0 equiv.) was added, and the mixture was stirred at 0 °C for 15 minutes. Under ice bath conditions, 4-chloropyrimidine (39.40 mg, 0.34 mmol, 1.1 equiv.) was added dropwise and the mixture was stirred for 1 hour. After monitoring the completion of the reaction by LC-MS, the mixture was extracted with water and dichloromethane and concentrated to obtain the crude title compound (112 mg, yield: 90%). LC-MS (ESI) [M + H] + = 398.2。

[0305] Step 5: Preparation of 2-(piperidin-1-yl)-6-((1-(pyrimidin-4-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid

Chemical formula

[0306] Methyl 2-(piperidin-1-yl)-6-((1-(pyrimidin-4-yl)piperidin-4-yl)amino)pyrimidine-4-carboxylate (112 mg, 0.28 mmol, 1.0 equiv.) was dissolved in acetonitrile (1 mL), potassium trimethylsilanolate (53.76 mg, 0.42 mmol, 1.5 equiv.) was added, and the mixture was stirred at 25 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, the mixture was concentrated and the pH was adjusted to weakly acidic with dilute hydrochloric acid solution to obtain the crude title compound (62 mg, yield 57.4%). LC-MS (ESI) [M + H] + = 385.1。

[0307] Production Example 30: 6-((2-(Methylsulfonyl)ethyl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 322).

[0308] Step 1: Preparation of 2-(Methylsulfonyl)ethan-1-amine

Chemical formula

[0309] 2-(Methylthio)ethylamine (500 mg, 5.48 mmol, 1.0 equiv.) was dissolved in dichloromethane (30 mL), m-chloroperoxybenzoic acid (2.08 g, 12.06 mmol, 2.2 equiv.) was added, and the mixture was stirred at room temperature (25 °C) for 3 hours, quenched with saturated sodium bisulfite solution, and the solvent was centrifuged to obtain a crude product (500 mg) of the title compound.

[0310] Step 2: Preparation of Methyl 2-chloro-6-((2-(methylsulfonyl)ethyl)amino)pyrimidine-4-carboxylate

Chemical formula

[0311] Methyl 2,6-dichloropyrimidine-4-carboxylate (1.2 g, 5.79 mmol, 1.0 equiv.) was dissolved in acetonitrile (25 mL), 2-(methylsulfonyl)ethan-1-amine (714 mg, 5.79 mmol, 1.0 equiv.) and N,N-diisopropylethylamine (2.25 g, 17.39 mmol, 3.0 equiv.) were added at 0 °C. The mixture was stirred at 0 °C for 1 hour under nitrogen. The completion of the reaction was monitored by TLC, the solvent was concentrated, dried on a rotary evaporator, and pulped with ethyl acetate, and the solid was collected by filtration to obtain the title compound (500 mg, yield 29.4%). LC-MS (ESI) [M + H] + = 294.0.

[0312] Step 3: Preparation of Methyl 6-((2-(Methylsulfonyl)ethyl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylate

Chemical formula

[0313] Methyl 2-chloro-6-((2-(methylsulfonyl)ethyl)amino)pyrimidine-4-carboxylate (250 mg, 0.85 mmol, 1.0 equiv.), tetrahydropyrrole (67 mg, 0.94 mmol, 1.1 equiv.) were dissolved in acetonitrile (5 mL), N,N-diisopropylethylamine (165 mg, 1.28 mmol, 1.5 equiv.) was added, and the mixture was stirred at 90 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, it was extracted three times with ethyl acetate (5 mL) and separated and purified by flash chromatography (silica gel, DCM:MeOH = 100:6) to obtain the title compound (140 mg, yield 50.1%). LC-MS (ESI) [M + H] + = 329.2

[0314] Step 4: Preparation of 6-((2-(Methylsulfonyl)ethyl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylic Acid

Chemical formula

[0315] Methyl 6-((2-(methylsulfonyl)ethyl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylate (150 mg, 0.46 mmol, 1.0 equiv.) was dissolved in tetrahydrofuran (2 mL), an aqueous lithium hydroxide solution (2 M) (21 mg, 0.91 mmol, 2.0 equiv.) was added, and the mixture was stirred at 25 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, it was concentrated to obtain the crude product of the title compound (170 mg). LC-MS (ESI) [M + H]+ = 315.2。

[0316] Production Example 31: 6 - ((2 - (Benzyloxycarbonyl)-2 - azaspiro[3.3]heptan - 6 - yl)amino)-2 - (pyrrolidin - 1 - yl)pyrimidine - 4 - carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 327).

[0317] Step 1: Preparation of Benzyl 6 - ((tert - butoxycarbonyl)amino)-2 - azaspiro[3.3]heptane - 2 - carboxylate

Chemical formula

[0318] tert - Butyl (2 - azaspiro[3.3]heptan - 6 - yl)carbamate (550 mg, 2.59 mmol, 1.0 equiv.), CbzCl (486 mg, 2.85 mmol, 1.1 equiv.), and DIPEA (670 mg, 5.18 mmol, 2.0 equiv.) were dissolved in DCM (13 mL) and reacted at 20°C for 3 hours. The reaction mixture was concentrated, and the crude product was separated and purified by flash chromatography (silica gel, DCM:MeOH = 49:1) to obtain the title compound (658 mg, yield 73.3%). LC - MS (ESI)[M + H] + = 347.2。

[0319] Step 2: Preparation of Benzyl 6 - amino - 2 - azaspiro[3.3]heptane - 2 - carboxylate

Chemical formula

[0320] Benzyl 6-((tert-butoxycarbonyl)amino)-2-azaspiro[3.3]heptane-2-carboxylate (658 mg, 1.90 mmol, 1.0 equiv.) and TFA (2.5 mL) were dissolved in dichloromethane (10 mL) and reacted at 20 °C for 1 hour. The reaction mixture was quenched with saturated aqueous sodium bicarbonate and extracted with dichloromethane. The organic phase was concentrated to obtain the crude product of the target compound (468 mg, yield 100%). LC-MS(ESI)[ M + H] + = 247.2。

[0321] Step 3: Preparation of Benzyl 6-((2-chloro-6-(methoxycarbonyl)pyrimidin-4-yl)amino)-2-azaspiro[3.3]heptane-2-carboxylate

Chem.

[0322] Benzyl 6-amino-2-azaspiro[3.3]heptane-2-carboxylate (468 mg, 1.90 mmol, 1.0 equiv.), methyl 2,6-dichloropyrimidine-4-carboxylate (393 mg, 1.90 mmol, 1.0 equiv.) and DIPEA (491 mg, 3.80 mmol, 2.0 equiv.) were dissolved in acetonitrile (20 mL) and reacted at 0 °C for 1 hour. The reaction mixture was concentrated and the crude product was separated and purified by flash chromatography (silica gel, DCM:MeOH = 49:1) to obtain the title compound (754 mg, yield 95.3%). LC-MS(ESI)[ M + H] + = 417.2。

[0323] Step 4: Preparation of Benzyl 6-((6-(methoxycarbonyl)-2-(pyrrolidin-1-yl)pyrimidin-4-yl)amino)-2-azaspiro[3.3]heptane-2-carboxylate

Chem.

[0324] Benzyl 6-((2-chloro-6-(methoxycarbonyl)pyrimidin-4-yl)amino)-2-azaspiro[3.3]heptane-2-carboxylate (120 mg, 0.29 mmol, 1.0 equiv.), tetrahydropyrrole (23 mg, 0.32 mmol, 1.1 equiv.) and DIPEA (74 mg, 0.58 mmol, 2.0 equiv.) were dissolved in acetonitrile (4 mL) and reacted at 90 °C for 24 h. The reaction mixture was concentrated and the crude product was separated and purified by flash chromatography (silica gel, DCM:MeOH = 49:1) to give the title compound (120 mg, yield 92.3%). LC-MS(ESI)[ M + H] + = 452.2。

[0325] Step 5: Preparation of 6-((2-(benzyloxycarbonyl)-2-azaspiro[3.3]heptan-6-yl)amino)-2-(pyrrolidin-1-yl)pyrimidine-4-carboxylic acid

Chemical formula

[0326] Benzyl 6-((6-(methoxycarbonyl)-2-(pyrrolidin-1-yl)pyrimidin-4-yl)amino)-2-azaspiro[3.3]heptane-2-carboxylate (120 mg, 0.27 mmol, 1.0 equiv.) and LiOH (13 mg, 0.53 mmol, 2.0 equiv.) were dissolved in THF (4 mL) and water (0.4 mL) and reacted at 20 °C for 1.5 h. The reaction solution was adjusted to acidic with dilute hydrochloric acid and concentrated to give the crude product (197 mg) of the target compound. LC-MS(ESI)[ M + H] + = 438.2。

[0327] Production Example 32: 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-(trifluoromethyl)pyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 356)

[0328] Step 1: Preparation of tert-Butyl 4-((6-Chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)piperidine-1-carboxylate

Chemical formula

[0329] 4,6-Dichloro-2-(trifluoromethyl)pyrimidine (2 g, 9.2 mmol), tert-Butyl 4-aminopiperidine-1-carboxylate (1.846 g, 9.2 mmol) and DIPEA (1.78 g, 13.82 mmol) were dissolved in acetonitrile (20 mL) and stirred at room temperature (25 - 30 °C) for 2 hours. The reaction was monitored by LC-MS and the raw materials disappeared. Ethyl acetate was extracted 3 times with 40 mL each time. The ethyl acetate phases were combined, washed once with 40 mL of water, once with 40 mL of saturated brine, dried over anhydrous sodium sulfate for 10 minutes, filtered, and the crude product was separated and purified by column chromatography using DCM:MeOH = 50:1 to obtain the product (3.47 g, yield 96.9%). LC-MS (ESI) [M + H] + = 381.2.

[0330] Step 2: Preparation of Methyl 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-(trifluoromethyl)pyrimidine-4-carboxylate

Chemical formula

[0331] tert-Butyl 4-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)piperidine-1-carboxylate (3.47 g, 9.1 mmol), Et3N (2.76 g, 27.3 mmol) and Pd(dppf)Cl2 (0.66 g, 0.91 mmol) were dissolved in a mixed solution of methanol (50 mL) and DMF (50 mL), and the reaction mixture was heated at 100 °C for 72 h in a 4 atm CO atmosphere. Completion of the conversion to the product was detected by LC-MS. The reaction solvent was dried on a rotary evaporator, extracted three times with 40 mL each of ethyl acetate, the ethyl acetate phases were combined, washed once with 40 mL of water, washed once with 40 mL of saturated brine, dried over anhydrous sodium sulfate for 10 min, filtered, and the crude product was separated and purified by column chromatography using DCM:MeOH = 30:1 to obtain the crude product (3 g, yield 81.6%). LC-MS(ESI)[ M + H] + = 405.2。

[0332] Step 3: Preparation of 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-(trifluoromethyl)pyrimidine-4-carboxylic acid

Chemical formula

[0333] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-(trifluoromethyl)pyrimidine-4-carboxylate (2 g, 4.95 mmol) was dissolved in THF (10 mL) and methanol (10 mL), and then an aqueous lithium hydroxide solution (5 mL, 10 mmol, 2M) was added. The reaction mixture was stirred at room temperature for 1 h. Completion of the reaction was monitored by LC-MS. The reaction mixture was extracted three times with 10 mL each of ethyl acetate. The aqueous phase was left, 1M hydrochloric acid was added to adjust the pH to 6 - 7, and then dried directly on a rotary evaporator to obtain the crude product (2.18 g) of the title compound. LC-MS(ESI)[ M + H] + = 391.2。

[0334] Production Example 33: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-cyclohexylpyrimidine-4-carboxylic acid (this intermediate can be used, for example, as an intermediate for the compound of Example 357)

[0335] Step 1: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(cyclohex-1-en-1-yl)pyrimidine-4-carboxylate

Chemical formula

[0336] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (300 mg, 0.959 mmol, 1 equiv.), cyclohex-1-en-1-ylboronic acid (241.6 mg, 1.918 mmol, 2.0 equiv.), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (70.24 mg, 0.096 mmol, 0.1 equiv.) and potassium carbonate (397.7 mg, 2.978 mmol, 3 equiv.) were weighed and mixed in dioxane (80 mL) and water (20 mL, pre-deoxygenated with nitrogen gas), and heated to 100 °C in an oil bath and reacted for 1 hour. The consumption of the reactants was monitored by LC-MS. The reaction solution was filtered and dried on a rotary evaporator to obtain a crude product (343 mg) of the title compound. LC-MS (ESI) [M + H] + = 359.2.

[0337] Step 2: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(cyclohex-1-en-1-yl)pyrimidine-4-carboxylic acid

Chemical formula

[0338] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(cyclohex-1-en-1-yl)pyrimidine-4-carboxylate (343 mg, 0.957 mmol, 1.0 equiv.) was added to water (1 mL) and tetrahydrofuran (4.0 mL) at room temperature (25 °C), lithium hydroxide (45.8 mg, 1.914 mmol, 2.0 equiv.) was added, and the mixture was stirred at room temperature for 1 hour. After monitoring the completion of the reaction by LC-MS, the reaction solution was concentrated to remove the organic solvent, adjusted to pH = 3 with 1 M hydrochloric acid, and the reaction system was extracted with dichloromethane:methanol (V / V = 10 / 1) (30 mL × 5). The organic phases were combined, dried, filtered, and concentrated to obtain the crude product (1.1 g) of the title compound. LC-MS (ESI) [M + H] + = 345.2.

[0339] Step 3: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-cyclohexylpyrimidine-4-carboxylic acid

Chemical Structure

[0340] 6-((1-acetylpiperidin-4-yl)amino)-2-(cyclohex-1-en-1-yl)pyrimidine-4-carboxylic acid (1.05 g, 3.049 mmol, 1.0 equiv.) was added to methanol (10 mL), and then palladium on carbon (324 mg, 0.305 mmol, 0.1 equiv., 10%) was added. The mixture was stirred under hydrogen at room temperature for 2 hours. After monitoring the completion of the reaction by LC-MS, the reaction system was filtered and concentrated to obtain the crude product, which was purified by Prep-HPLC (aqueous NH4NH3, MeCN) 10 mmol / L and lyophilized to obtain the title compound (148 mg, yield 14%). LC-MS (ESI) [M + H] + = 347.3.

[0341] Production Example 34: 6-((1-Acetylpiperidin-4-yl)amino)-2-ethylpyrimidine-4-carboxylic acid (This intermediate can be used, for example, as an intermediate for the compound of Example 365)

[0342] Step 1: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-((trimethylsilyl)ethynyl)pyrimidine-4-carboxylate

Chemical formula

[0343] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (999.86 mg, 3.2 mmol, 1 equiv.), ethynyltrimethylsilane (1.88 g, 0.019 mol, 6 equiv.), copper(I) iodide (0.091 g, 0.48 mmol, 0.15 equiv.), triethylamine (0.97 g, 9.6 mmol, 3 equiv.) and tetrakis(triphenylphosphine)palladium(0) (0.55 g, 0.48 mmol, 0.15 equiv.) were dissolved in DMF (N,N-dimethylformamide) (8 mL), purged with nitrogen for 2 minutes, and reacted by microwave heating at 100 °C for 2 hours. After monitoring the completion of the reaction by TLC, the mixture was filtered, and the cake was washed twice with 15 mL of a mixed solvent (DCM:MeOH = 10:1). The filtrate was concentrated, and the crude product was separated by column chromatography (DCM:MeOH = 10:1) and concentrated to obtain the title compound (0.5 g, yield 41.8%).

[0344] 11H NMR (400 MHz, DMSO) δ 7.95 (d2, J = 7.4 Hz, 1H), 7.06 (s, 1H), 4.17 (d, J = 14.0 Hz, 2H), 3.84 (s, 3H), 3.75 (s, 1H), 3.22 (t, J = 11.7 Hz, 1H), 2.86 (t, J = 10.9 Hz, 1H), 2.01 (s, 3H), 1.87 (dd, J = 23.0 Hz, 12.0 Hz,2H), 1.42-1.33 (m, 1H),1.33-1.25 (m, 1H), 0.26 (s, 9H).

[0345] Step 2: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-ethynylpyrimidine-4-carboxylate

Chemical Structure

[0346] A solution of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-((trimethylsilyl)ethynyl)pyrimidine-4-carboxylate (0.2 g, 0.53 mmol, 1 equiv.) in tetrahydrofuran (10 mL) was added with a tetrahydrofuran solution of tetrabutylammonium fluoride (0.21 mL, 0.80 mmol, 1.5 equiv.). The reaction was carried out at 25 °C for half an hour. After monitoring the completion of the reaction by LC-MS, it was concentrated to obtain a crude product (161.44 mg, 0.534 mmol), which was used directly in the next reaction without purification. LC-MS (ESI) [M + H] + = 303.1.

[0347] Step 3: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-ethylpyrimidine-4-carboxylate

Chemical Structure

[0348] Pd / C (0.0227 g, 0.11 mmol, 0.2 equiv.) was added to a solution of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-ethynylpyrimidine-4-carboxylate (161.44 mg, 0.53 mmol, 1 equiv.) in methanol (10 mL) under a nitrogen atmosphere. After four replacements with hydrogen, reaction at 25 °C for 8 h, and monitoring the completion of the reaction by LC-MS, the crude product was obtained by washing the filter cake three times with methanol (10 mL) and concentrating the filtrate, and was purified by reverse phase (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain the title compound (60 mg, yield: 36.7%). LC-MS (ESI) [M + H] + = 307.2。

[0349] Step 4: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-ethylpyrimidine-4-carboxylic acid

Chemical Structure

[0350] To a mixed solution of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-ethylpyrimidine-4-carboxylate (60 mg, 0.20 mmol, 1 equiv.) in tetrahydrofuran (3 mL, 33.33%) and methanol (3 mL, 33.33%), an aqueous solution (3 mL, 33.33%) of lithium hydroxide (20.54 mg, 0.49 mmol, 2.5 equiv.) was added. The reaction was carried out at 25 °C for 1 h, and after monitoring the completion of the reaction by LC-MS, it was concentrated, the crude product was dissolved in 2 mL of acetonitrile, the pH was adjusted to 5 with 1M aqueous HCl, and freeze-dried to obtain the title compound (50 mg, yield 87.3%). LC-MS (ESI) [M + H] + = 293.2。

[0351] Preparation of trans-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone

[0352] Step 1: Preparation of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylate

Chemical formula

[0353] Methyl 6-chloropyrimidine-4-carboxylate (1 g, 5.80 mmol, 1.0 equiv.), tert-butyl 4-aminopiperidine-1-carboxylate hydrochloride (1.65 g, 6.95 mmol, 1.2 equiv.) and DIPEA (N,N-diisopropylethylamine) (3.00 g, 23.18 mmol, 4.0 equiv.) were dissolved in MeCN (acetonitrile) (30 mL) and reacted at 90 °C for 15 hours. The reaction solution was concentrated, and the crude product was separated and purified by flash chromatography (silica gel, PE:EA = 3:2) to obtain the target compound (1.82 g, 93.4%). LC-MS (ESI) [M + H] + = 337.3.

[0354] Step 2: Preparation of 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid

Chemical formula

[0355] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylate (1.8 g, 5.35 mmol, 1.0 equiv.) and LiOH (lithium hydroxide) (0.26 g, 10.70 mmol, 2.0 equiv.) were dissolved in THF (tetrahydrofuran) (27 mL) and H2O (water) (9 mL), and reacted at 20 °C for 1 hour. The reaction solution was concentrated, diluted with dichloromethane / methanol (3:1), suction filtered, and the filtrate was concentrated to obtain a crude product of the target compound (2.14 g, yield not calculated). LC-MS (ESI) [M + H] + = 323.2。

[0356] Step 3: Preparation of tert-butyl 6-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidine-1-carboxylate

Chemical formula

[0357] 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)pyrimidine-4-carboxylic acid (2.1 g, 6.51 mmol, 1.0 equiv.), trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (2.09 g, 8.99 mmol, 1.38 equiv.), ethyl acetate solution of T3P (1-N-propylphosphoric anhydride) (mass fraction 50%) (8.29 g, 13.03 mmol, 2.0 equiv.) and TEA (triethylamine) (3.30 g, 32.57 mmol, 5.0 equiv.) were dissolved in DMF (N,N-dimethylformamide) (30 mL) and reacted at 20 °C for 17 h. The reaction mixture was extracted with ethyl acetate, the organic phase was concentrated, and the crude product was separated and purified by flash chromatography (silica gel, DCM:MeOH = 20:1) to obtain the crude title compound (0.98 g, 28.0%). 30 mg of the crude target compound was obtained and separated and purified by Prep-HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain the target compound (19.4 mg).

[0358] LC-MS (ESI) [M + H] + = 537.6, 1 1H NMR (400 MHz, CDCl3) δ 8.57-8.51 (m, 1H), 7.19-7.08 (m, 3H), 7.06-6.99 (m, 1H), 6.63-6.54 (m, 1H), 5.17-4.66 (m, 2H), 4.32-3.84 (m, 6H), 3.77-3.62 (m, 2H), 3.17-2.80 (m, 7H), 2.79-2.54 (m, 2H), 2.08-1.81 (m, 3H), 1.76-1.65 (m, 1H), 1.47 (s, 9H), 1.44-1.35 (m, 2H).

[0359] Step 4: Preparation of trans-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone [Chemistry]

[0360] tert-Butyl trans-4-((6-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidine-1-carboxylate (30 mg, 0.06 mmol, 1.0 equiv.) was dissolved in DCM (dichloromethane) (0.4 mL), and then TFA (trifluoroacetic acid) (0.1 mL) was added, and the reaction was carried out at 20 °C for 0.5 h. The reaction mixture was quenched with saturated aqueous sodium bicarbonate solution, concentrated, and the crude product was separated and purified by preparative HPLC (aqueous C18, 10 mmol / L NH4NH3 / acetonitrile) to obtain the title compound (17.42 mg, 71.4%).

[0361] LC-MS (ESI) [M + H] + = 437.5; LC-MS (ESI) [M+H] + = 437.5; 1 H NMR (400 MHz, DMSO-d6) δ 8.46 - 8.32 (m, 1H), 7.61 (t, J = 7.7 Hz, 1H), 7.13 - 7.01 (m, 4H), 6.51 (s, 1H), 4.79 (dd, J = 35.1, 3.9 Hz, 1H), 4.40 (dd, J = 56.4, 12.7 Hz, 1H), 3.93 (m, 1H), 3.87 - 3.56 (m, 5H), 2.98 (d, J = 12.5 Hz, 2H), 2.93 - 2.87 (m, 1H), 2.85 - 2.75 (m, 4H), 2.69 - 2.53 (m, 4H), 1.92 - 1.69 (m, 3H), 1.57 - 1.43 (m, 1H), 1.41 - 1.27 (m, 2H).

[0362] Production Example 36: Preparation of (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl-4,4-d2)piperidin-3-ol

[0363] Step 1: Preparation of tert-butyl (3R,4R)-3-((tert-butoxycarbonyl)oxy)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate

Chem.

[0364] (3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (1 g, 4.3 mmol, 1.0 equiv.), TEA (1.3 g, 12.9 mmol, 3.0 equiv.) and DMAP (105 mg, 0.86 mmol, 0.2 equiv.) were dissolved in THF (20 mL), and Boc2O (1.88 g, 8.6 mmol, 2.0 equiv.) was added while stirring, and the reaction was carried out at 20 °C for 1 hour. The reaction solution was concentrated, and the crude product was separated and purified by column chromatography (silica gel, PE:EA = 20:1) to obtain the target product (1.12 g, yield 60%). LC-MS (ESI) [M + H] + = 433.3。

[0365] Step 2: Preparation of Tert-butyl (3R,4R)-3-((tert-butoxycarbonyl)oxy)-4-(4-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate

Chem.

[0366] tert-Butyl (3R,4R)-3-((tert-butoxycarbonyl)oxy)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate (300 mg, 0.49 mmol, 1.0 equiv.) was dissolved in CHCl3 (10 mL), and formic acid (451 mg, 9.8 mmol, 20.0 equiv.) and DDQ (473 mg, 2.08 mmol, 3.0 equiv.) were added. The reaction was carried out at 20 °C for 16 h. Saturated aqueous sodium carbonate solution (5 mL) was added to the reaction mixture, and the mixture was extracted with water and DCM. The organic phase was concentrated, and the crude product was separated and purified by column chromatography (silica gel, DCM:MeOH = 50:1) to obtain the target compound (46 mg, yield 18%). LC-MS(ESI)[ M + Na] + = 469.3。

[0367] Step 3: Preparation of 2-(3R,4R-3-hydroxypiperidin-4-yl)-2,3-dihydroisoquinolin-4(1H)-one

Chemical Structure

[0368] tert-Butyl (3R,4R)-3-((tert-butoxycarbonyl)oxy)-4-(4-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate (46 mg, 0.1 mmol, 1.0 equiv.) was dissolved in EA (0.5 mL), and ethyl acetate-hydrogen chloride gas (5 mL, 4M) was added. The reaction was carried out at 20 °C for 1 h. The reaction solution was concentrated, and the crude product was dissolved in MeOH:H2O (1 mL:1 mL). Lithium hydroxide (7 mg, 0.3 mmol, 3.0 equiv.) was added, and the reaction was carried out at 20 °C for 1 h. The reaction mixture was concentrated and purified by pre-TL (silica gel, DCM:MeOH = 5:1) to obtain the target compound (12 mg, yield 48%). LC-MS(ESI)[ M + H] + = 247.3。

[0369] Step 4: Preparation of (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl-4,4-d2)piperidin-3-ol

Chem.

[0370] 2-((3R,4R)-3-Hydroxypiperidin-4-yl)-2,3-dihydroisoquinolin-4(1H)-one (4 mg, 0.016 mmol, 1.0 equiv.) was dissolved in dry THF (1 mL), LiAlD4 (1.3 mg, 0.032 mmol, 2.0 equiv.) was added, and the reaction was carried out at 0 °C for 2 hours. The reaction solution was quenched with heavy water and concentrated to obtain the target compound (6 mg, crude product). LC-MS (ESI) [M + H] + = 235.1。

[0371] Production Example 37: Preparation of 2-(3R,4R-3-hydroxypiperidin-4-yl)-1,2,3,4-tetrahydroisoquinolin-4-ol

[0372] Step 1: Preparation of tert-butyl (3R,4R)-3-((tert-butoxycarbonyl)oxy)-4-(4-hydroxy-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate

Chem.

[0373] tert-Butyl (3R,4R)-3-((tert-butoxycarbonyl)oxy)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate (500 mg, 1.16 mmol, 1.0 equiv.) was dissolved in a mixed solution of THF:H2O (10 mL:10 mL), and Oxone (1.42 g, 2.31 mmol, 2.0 equiv.) was added with stirring, followed by reaction at 20 °C for 16 h. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL × 3). The combined organic phases were dried and concentrated, and the crude product was separated and purified by column chromatography (silica gel, DCM:MeOH = 50:1) to obtain the title compound (120 mg, yield 23%). LC-MS(ESI)[ M + H] + = 449.3。

[0374] Step 2: Preparation of 2-(3R,4R-3-hydroxypiperidin-4-yl)-1,2,3,4-tetrahydroisoquinolin-4-ol

Chemical Structure

[0375] tert-Butyl (3R,4R)-3-((tert-butoxycarbonyl)oxy)-4-(4-hydroxy-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate (100 mg, 0.223 mmol, 1.0 equiv.) was dissolved in EA (0.5 mL), and ethyl acetate hydrochloric acid gas (5 mL, 4M) was added, followed by reaction at 20 °C for 1 h. The reaction solution was concentrated, and the crude product was dissolved in MeOH:H2O (1 mL:1 mL), and lithium hydroxide monohydrate (29 mg, 0.669 mmol, 3.0 equiv.) was added, followed by reaction at 20 °C for 1 h. The reaction mixture was concentrated and purified by pre-TL (silica gel, DCM:MeOH = 5:1) to obtain the target compound (35 mg, yield 64%). LC-MS(ESI)[ M + H] + = 249.2。

[0376] Production Example 38: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(tert-butylthio)pyrimidine-4-carboxylic acid

[0377] Step 1: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(tert-butylthio)pyrimidine-4-carboxylate

Chemical formula

[0378] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (1 g, 3.20 mmol, 1 equiv.), potassium carbonate (0.884 g, 6.40 mmol, 2 equiv.) were added to N,N-dimethylformamide (16 mL), and further t-butyl mercaptan (1.45 g, 16.0 mmol, 5 equiv.) was added. The mixture was stirred at 80 °C for 6 hours, and it was detected by TLC that the reaction of the starting material was complete. Water (160 mL) was added to the reaction system, and the mixture was extracted 3 times with ethyl acetate (3 × 50 mL). The organic phases were combined, washed once with a saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered by suction, the solvent was concentrated, and the crude product was purified by column chromatography (DCM:MeOH = 98:2) to obtain the title compound (0.49 g, yield 41.8%). LC-MS (ESI) [M + H] + = 367.2.

[0379] Step 2: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(tert-butylthio)pyrimidine-4-carboxylic acid

Chemical formula

[0380] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(tert-butylthio)pyrimidine-4-carboxylate (0.29 g, 0.791 mmol, 1 equiv.) was dissolved in tetrahydrofuran / water (25 mL, 4:1), lithium hydroxide (38 mg, 1.582 mmol, 2 equiv.) was added, and the mixture was stirred at 16 °C for 2 h. The completion of the reaction was monitored by TLC. Water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The aqueous phase was adjusted to pH = 4 - 5 with 1 M HCl and lyophilized to obtain the crude product of the title compound (0.25 g, yield 89.7%). LC-MS(ESI)[ M + H] + = 353.2。

[0381] Production Example 39: Preparation of methyl 6-(((1-acetylazetidin-3-yl)amino)-2-chloropyrimidine-4-carboxylate

[0382] Step 1: Preparation of methyl 6-(((1-acetylazetidin-3-yl)amino)-2-chloropyrimidine-4-carboxylate

Chemical formula

[0383] Methyl 2,6-dichloropyrimidine-4-carboxylate (500 mg, 2.42 mmol, 1.2 equiv.), 1-(3-aminoazetidin-1-yl)ethan-1-one (230 mg, 2.02 mmol, 1.0 equiv.) were dissolved in acetonitrile (5 mL), N,N-diisopropylethylamine (521 mg, 4.04 mmol, 2.0 equiv.) was added, and the mixture was reacted at room temperature for 2 h. After monitoring the completion of the reaction by LC-MS, the solvent was dried by rotary evaporator, and the crude product was separated and purified by flash chromatography (silica gel, DCM:MeOH = 5%) to obtain the title compound (530 mg, yield 92%). LC-MS(ESI)[ M + H] + = 285.1。

[0384] Example 1: Preparation of trans-1-(4-((6-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)oxy)piperidin-1-yl)ethan-1-one

Chemical Structure

[0385] 1-(4-Hydroxypiperidin-1-yl)ethan-1-one (77 mg, 0.536 mmol, 2.0 equiv.) was added to a solution of NaH (60% by mass) (35.7 mg, 0.536 mmol, 2.0 equiv.) in THF (1.4 mL), and the mixture was stirred at 0 °C under nitrogen protection for 0.5 h. Then, trans-((6-chloropyrimidin-4-yl)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone (100 mg, 0.268 mmol, 1.0 equiv.) was added, and the stirring was continued at 16 °C for 0.5 h. The completion of the reaction was monitored by LC-MS. 10 mL of saturated ammonium chloride solution was added dropwise at 0 °C, and the mixture was extracted once with EA (20 mL). The organic phase was washed once with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered by suction, the mother liquor was concentrated, and the crude product was purified by reverse-phase separation (C18, 10 mmol / L ammonium bicarbonate / acetonitrile) to obtain the title compound (2.32 mg, yield: 1.5%).

[0386] LC-MS (ESI): m / z = 480.5; 11H NMR (400 MHz, CDCl3) δ 8.76 - 8.74 (m, 1H), 7.21 - 7.10 (m, 3H), 7.06 - 7.01 (m, 1H), 7.00 - 6.93 (m, 1H), 5.49 - 5.37 (m, 1H), 5.09 - 4.76 (m, 1H), 4.18 - 4.06 (m, 1H), 4.03 - 3.92 (m, 2H), 3.79 - 3.68 (m, 3H), 3.56 - 3.39 (m, 2H), 3.13 - 2.92 (m, 4H), 2.83 - 2.72 (m, 2H), 2.14 (s, 3H), 2.10 - 2.02 (m, 2H), 1.86 - 1.67 (m, 6H).

[0387] Examples 2 and 3: Preparation of (Z)-2-(4-((6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidine-1-carbonyl)-3-(dimethylamino)acrylonitrile (Compound of Example 2) and (3-(4-((6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropionitrile (Compound of Example 3)

[0388] Step 1: Preparation of (Z)-2-cyano-3-(dimethylamino)acryloyl chloride and 2-cyanoacetyl chloride

Chem.

[0389] Cyanoacetic acid (1 g, 11.8 mmol, 1.0 equiv.) was dissolved in DCM (100 mL), and (COCl)2 (1.64 g, 12.9 mmol, 1.1 equiv.) and DMF (8 mg, 0.1 mmol, 0.01 equiv.) were added at 0 °C. The mixture was stirred at 25 °C for 1 h under nitrogen until the reaction was completed by TLC detection, and then dried by rotary evaporator to obtain a mixture (1.1 g, crude) of (Z)-2-cyano-3-(dimethylamino)acryloyl chloride and 2-cyanoacetyl chloride.

[0390] Step 2: Preparation of (Z)-2-(4-((6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidine-1-carbonyl)-3-(dimethylamino)acrylonitrile (Example 2) and 3-(4-((6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropionitrile (Example 3) [Chemical formula]

[0391] A mixture of (Z)-2-cyano-3-(dimethylamino)acryloyl chloride and 2-cyanoacetyl chloride (5 mg, 0.046 mmol, 1.0 equiv.) was dissolved in DCM (2 mL), and a DCM solution (1.5 mL) of (trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone (20 mg, 0.046 mmol, 1.0 equiv.) was added dropwise at 0 °C. Triethylamine (9 mg, 0.09 mmol, 2.0 equiv.) was added, and the mixture was stirred at 0 °C for 1 h. After monitoring the completion of the reaction by LC-MS, the mixture was concentrated and the crude product was purified by reverse-phase separation (C18, 10 mmol / L aqueous ammonium bicarbonate / acetonitrile) to obtain the two title compounds:

[0392] Compound of Example 2 (14.5 mg, yield 56.7%). LC-MS (ESI) [M+H] + = 559.56; 1 1H NMR (400 MHz, CDCl3) δ 8.60-8.48 (m, 1H), 7.54 (s, 1H), 7.19-7.09 (m, 3H), 7.08-6.98 (m, 1H), 6.67-6.55 (m, 1H), 5.47-5.27 (m, 1H), 5.07-4.63 (m, 1H), 4.31-4.23 (m, 2H), 4.22-4.12 (m, 1H), 4.04-3.91 (m, 2H), 3.79-3.68 (m, 2H), 3.35 (s, 3H), 3.17 (s, 3H), 3.12-2.98 (m, 4H), 2.97-2.83 (m, 3H), 2.76-2.65 (m, 2H), 2.13-2.05 (m, 2H), 2.03-1.89 (m, 1H), 1.71-1.64 (m, 1H), 1.60-1.43 (m, 2H).

[0393] Compound of Example 3 (5.8 mg, yield: 25.4%). LC-MS (ESI) [M+H] + = 504.56; 11H NMR (400 MHz, CDCl3) δ 8.60-8.45 (m, 1H), 7.20-7.09 (m, 3H), 7.08-6.99 (m, 1H), 6.71-6.58 (m, 1H), 6.02-5.60 (m, 1H), 5.07-4.65 (m, 1H), 4.47 (d, J = 13.3 Hz, 1H), 4.23-4.10 (m, 2H), 4.01-3.93 (m, 1H), 3.79-3.69 (m, 3H), 3.63-3.46 (m, 2H), 3.37-3.26 (m, 1H), 3.11-2.99 (m, 2H), 2.97-2.88 (m, 3H), 2.80-2.66 (m, 2H), 2.20-2.04 (m, 2H), 2.01-1.78 (m, 3H), 1.76-1.63 (m, 1H), 1.61-1.41 (m, 2H).

[0394] Example 4: Preparation of trans-1-(6-((6-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)-2-azaspiro[3.3]heptan-2-yl)ethan-1-one

[0395] Step 1: Preparation of tert-butyl 2-acetyl-2-azaspiro[3.3]hept-6-yl)carbamate [Chemical formula]

[0396] tert-Butyl (2-azaspiro[3.3]heptan-6-yl) carbamate (300 mg, 1.41 mmol, 1.0 equiv.) and DIPEA (N,N-diisopropylethylamine) (365 mg, 2.83 mmol, 2.0 equiv.) were dissolved in DCM (dichloromethane) (7 mL), and then acetic anhydride (159 mg, 1.55 mmol, 1.1 equiv.) was added at 0 °C, and the reaction was carried out at 20 °C for 0.5 h. The reaction mixture was concentrated, and the crude product was separated and purified by flash chromatography (silica gel, DCM:MeOH = 30:1) to obtain the title compound (336 mg, yield 93.5%). LC-MS (ESI) [M + H] + = 255.2。

[0397] Step 2: Preparation of 1-(6-amino-2-azaspiro[3.3]heptan-2-yl)ethan-1-one

Chem.

[0398] tert-Butyl (2-acetyl-2-azaspiro[3.3]heptan-6-yl) carbamate (310 mg, 1.22 mmol, 1.0 equiv.) was dissolved in MeOH (methanol) (1 mL), and then a 1,4-dioxane solution of hydrochloric acid (3 mL, 4 M) was added, and the reaction was carried out at 20 °C for 1 h. The reaction solution was concentrated to obtain the crude product of the target compound (290 mg). LC-MS (ESI) [M + H] + = 155.1。

[0399] Step 3: Preparation of trans-1-(6-((6-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)-2-azaspiro[3.3]heptan-2-yl)ethan-1-one

Chem.

[0400] (6-Chloropyrimidin-4-yl)(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone (60 mg, 0.16 mmol, 1.0 equiv.), 1-(6-amino-2-azaspiro[3.3]heptan-2-yl)ethan-1-one (50 mg, 0.32 mmol, 2.0 equiv.), Pd(OAc)2 (palladium acetate) (7 mg, 0.03 mmol, 0.2 equiv.), BINAP (1,1'-binaphthalene-2,2'-bis(diphenylphosphine)) (40 mg, 0.06 mmol, 0.4 equiv.) and Cs2CO3 (cesium carbonate) (262 mg, 0.80 mmol, 5.0 equiv.) were dissolved in 1,4-dioxane (1 mL) and reacted at 80 °C for 16 h under nitrogen protection. The reaction mixture was filtered, the filtrate was concentrated, and the crude product was separated and purified by Prep-HPLC (aqueous NH4HO3, 10 mmol / L in MeCN) to obtain the title compound (2.04 mg, yield 2.6%).

[0401] LC-MS (ESI) [M+H] + = 491.5; 1 H NMR (400 MHz, CDCl3) δ 8.57 - 8.51 (m, 1H), 7.20 - 7.09 (m, 3H), 7.07 - 7.00 (m, 1H), 6.61 - 6.50 (m, 1H), 5.52 - 5.32 (m, 1H), 5.07 - 4.66 (m, 1H), 4.26 - 4.05 (m, 5H), 4.04 - 3.95 (m, 2H), 3.81 - 3.67 (m, 2H), 3.16 - 2.58 (m, 10H), 2.22 - 2.09 (m, 2H), 2.05 - 1.98 (m, 1H), 1.92 - 1.83 (m, 4H).

[0402] Example 5: Preparation of 1-(4-((2-(benzo[d]thiazol-7-yl)-6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0403] Step 1: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(benzo[d]thiazol-7-yl)pyrimidine-4-carboxylic acid

Chemical formula

[0404] 7-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazole (190 mg, 0.728 mmol, 1.0 equiv.), methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (249 mg, 0.8 mmol, 1.1 equiv.), Pd(dppf)Cl2 (53 mg, 0.0728 mmol, 0.1 equiv.) and potassium carbonate (301 mg, 2.18 mmol, 3.0 equiv.) were dissolved in dioxane (3 mL) and water (1 mL), and reacted at 100 °C for 1 hour. After completion of the reaction by LC-MS, it was diluted with water, extracted with EA, the aqueous phase was adjusted to pH 4 with 1M hydrochloric acid, and the aqueous phase was lyophilized to obtain a crude product (300 mg). LC-MS(ESI)[M + H] + = 398.3。

[0405] Step 2: Preparation of 1-(4-((2-(benzo[d]thiazol-7-yl)-6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chemical formula

[0406] 6-((1-Acetylpiperidin-4-yl)amino)-2-(benzothiazol-7-yl)pyrimidine-4-carboxylic acid (85 mg, 0.215 mmol, 1.0 equiv.), (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (50 mg, 0.215 mmol, 1.0 equiv.), EDCI (62 mg, 0.32 mmol, 1.5 equiv.), HOAt (44 mg, 0.32 mmol, 1.5 equiv.) were dissolved in DMF (2 mL) and reacted at 25 °C for 1 h. After monitoring the completion of the reaction by LC-MS, the reaction mixture was concentrated and separated by reverse-phase HPLC (C 18 , 10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain the title compound (26.28 mg, yield 20%).

[0407] LC-MS (ESI) [M+H] + = 612.2; 1 H NMR (400 MHz, CDCl3) δ 9.10 (s, 1H), 8.72 - 8.57 (m, 1H), 8.28 - 8.25 (m, 1H), 7.68 - 7.63 (m, 1H), 7.20 - 7.10 (m, 3H), 7.08 - 7.02 (m, 1H), 6.66 - 6.55 (m, 1H), 5.49 - 5.30 (m, 1H), 5.11 - 4.86 (m, 1H), 4.68 - 4.26 (m, 3H), 4.07 - 4.04 (m, 1H), 3.80 - 3.85 (m, 3H), 3.34 - 3.32 (m, 1H), 3.20 - 2.70 (m, 8H), 2.32 - 2.03 (m, 6H), 1.88 - 1.75 (m, 1H), 1.55 - 1.50 (m, 2H).

[0408] Example 6: Preparation of 1-(3-((6-(3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(thiazol-5-yl)pyrimidin-4-yl)amino)azetidin-1-yl)ethan-1-one

[0409] Step 1: Preparation of methyl 6-((1-acetylazetidin-3-yl)amino)-2-(thiazol-5-yl)pyrimidine-4-carboxylate

Chemical formula

[0410] Methyl 6-((1-acetylazetidin-3-yl)amino)-2-chloropyrimidine-4-carboxylate (50 mg, 0.176 mmol, 1.0 equiv.) was added to DMF (2.0 mL) at room temperature (25 °C), and 5-(tributylstannyl)thiazole (1.503 g, 0.351 mmol, 2.0 equiv.) and tetrakis(triphenylphosphine)palladium (20.3 mg, 0.018 mmol, 0.1 equiv.) were added under nitrogen protection. The mixture was stirred at 140 °C for 1 hour using microwave, and the completion of the reaction was monitored by LC-MS. Then it was filtered, the mother liquor was concentrated, and the crude product was separated and purified using a chromatography column (DCM:MeOH = 0 - 10%) to obtain the title compound (35 mg, yield: 59.7%). LC-MS (ESI) [M + H] + = 334.1。

[0411] Step 2: Preparation of 6-((1-acetylazetidin-3-yl)amino)-2-(thiazol-5-yl)pyrimidine-4-carboxylic acid

Chemical formula

[0412] Methyl 6-((1-acetylazetidin-3-yl)amino)-2-(thiazol-5-yl)pyrimidine-4-carboxylate (30 mg, 0.09 mmol, 1.0 equiv.) was added to water (1 mL) and tetrahydrofuran (2.0 mL) at room temperature, lithium hydroxide (4.3 mg, 0.18 mmol, 2.0 equiv.) was added, and the mixture was stirred at room temperature for 1 hour. The completion of the reaction was monitored by LC-MS, the organic solvent was concentrated and removed, the pH was adjusted to 3 with 1 M hydrochloric acid, the reaction system was extracted three times with dichloromethane:methanol (V / V = 10 / 1) (30 mL × 5), the organic phases were combined, dried, filtered, and concentrated to obtain the title compound (20 mg, crude product). LC-MS (ESI) [M + H] + = 320.3。

[0413] Step 3: Preparation of 1-(3-((6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(thiazol-5-yl)pyrimidine-4-yl)amino)azetidin-1-yl)ethan-1-one

Chemical formula

[0414] Using the method of the second stage of Example 5, the title compound (4.96 mg, yield: 14.8%) was synthesized. LC-MS (ESI) [M+H] + = 534.4; 1 H NMR (400 MHz, CD3OD) δ 9.06 (d, J = 6.0 Hz, 1H), 8.59 (d, J = 11.2 Hz, 1H), 7.10 - 7.06 (m, 4H), 6.58 (d, J = 8.0 Hz, 1H), 4.76 - 3.68 (m, 9H), 3.07 - 2.70 (m, 10H), 1.92 (s, 3H).

[0415] Example 7: Preparation of trans-1-(4-((6-(4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(thiazol-2-yl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0416] Step 1: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(thiazol-2-yl)pyrimidine-4-carboxylate

Chemical Structure

[0417] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (4.50 g, 1.44 mmol, 1 equiv.) and Pd(PPh3)4 (1.68 g, 0.144 mmol, 0.1 equiv.) were weighed and placed in a reaction flask. Dioxane (72 mL) and 2-(tributylstannyl)thiazole (9 mL, 2.88 mmol, 1.99 equiv.) were added, and the mixture was stirred at 110 °C for 14 h under nitrogen protection. The formation of the product was monitored by LC-MS. The solvent was concentrated, and the crude product was separated and purified by flash chromatography (silica gel, MeOH:EA = 0 - 5%) to obtain the title compound (1.11 g, yield 21.3%). LC-MS (ESI) [M + H] + = 362.2.

[0418] Step 2: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(thiazol-2-yl)pyrimidine-4-carboxylic acid

Chemical Structure

[0419] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(thiazol-2-yl)pyrimidine-4-carboxylate (1.11 g, 3.06 mmol, 1 equiv.) was dissolved in THF (24 mL), water (6 mL) and LiOH (180 mg, 7.5 mmol, 2.45 equiv.) were added, and the mixture was stirred at room temperature for 1 hour. The completion of the reaction was monitored by TLC. 1 M HCl was added to adjust the pH to 3 - 4, and the aqueous solution was lyophilized to obtain the crude product (1.0 g), which was used directly in the next step. LC-MS (ESI) [M + H] + = 348.0.

[0420] Step 3: Preparation of trans-1-(4-((6-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(thiazol-2-yl)pyrimidine-4-yl)amino)piperidin-1-yl)ethan-1-one

Chemical Structure

[0421] Using the method of the second step of Example 5, the title compound was synthesized (555 mg, yield 32%). LC-MS (ESI) [M+H] + = 562.5; 11H NMR (400 MHz, CDCl3) δ 8.04 - 7.95 (m, 1H), 7.52 - 7.46 (m, 1H), 7.20 - 7.10 (m, 3H), 7.07 - 7.01 (m, 1H), 6.75 - 6.62 (m, 1H), 5.81 - 5.54 (m, 1H), 5.13 - 4.66 (m, 1H), 4.49 (d, J = 13.6 Hz, 1H), 4.43 - 4.25 (m, 1H), 4.05 - 3.96 (m, 1H), 3.94 - 3.70 (m, 4H), 3.33 - 3.21 (m, 1H), 3.17 - 2.55 (m, 8H), 2.23 - 1.99 (m, 6H), 1.98 - 1.82 (m, 1H), 1.54 - 1.41 (m, 2H).

[0422] Example 8 - 138 The compounds of Examples 8 to 138 were synthesized using the same methods as in Examples 5, 6, and 7, and the compound structures and specific identification data (LC - MS and 1 1H NMR) were as follows: [Table 2 - 1] [Table 2 - 2] [Table 2 - 3] [Table 2 - 4] [Table 2 - 5] [Table 2 - 6] [Table 2 - 7] [Table 2 - 8] [Table 2 - 9]

Table 2-10

Table 2-11

Table 2-12

Table 2-13

Table 2-14

Table 2-15

Table 2-16

Table 2-17

Table 2-18

Table 2-19

[0423] Example 139: Preparation of trans-1-(3-((6-(3S,4S-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)phenyl)pyrrolidin-2-one

[0424] Step 1: Preparation of 1-(3-nitrophenyl)pyrrolidin-2-one

Chemical formula

[0425] To dimethyl sulfoxide (20 mL), (3-nitrophenyl)boronic acid (2.0 g, 18.6 mmol, 1.0 equiv) was added. Then, 2-pyrrolidone (4.74 g, 55.8 mmol, 3 equiv) and copper(I) iodide (178 mg, 0.93 mmol, 0.05 equiv) were added. After stirring at room temperature for 10 minutes, 70% aqueous t-butyl hydroperoxide (2.63 g, 20.46 mmol, 1.1 equiv) was added. After stirring at room temperature for 10 minutes, the reaction mixture was stirred at 60 °C for 4 hours. After monitoring the completion of the reaction by LC-MS, it was cooled to room temperature (28 °C), water (100 mL) was added, and the mixture was extracted three times with ethyl acetate (40 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate for 10 minutes, filtered, concentrated, and the crude product was separated and purified by flash chromatography (silica gel, petroleum ether / ethyl acetate (V / V = 4 / 1)) to obtain the title compound (800 mg, yield 20.87%). LC-MS (ESI) [ M + H] + = 207.1, 1 H NMR (400 MHz, MeOD) δ 8.72 (t, J = 2.4 Hz, 1H), 7.97 (t, J = 8.0 Hz, 2H), 7.68 (t, J = 8.0 Hz, 1H), 3.92 (t, J = 6.8 Hz, 2H), 2.57 (t, J = 8 Hz, 2H), 2.12~2.08 (m, 2H).

[0426] Step 2: Preparation of 1-(3-aminophenyl)pyrrolidin-2-one

Chemical Structure

[0427] At 28 °C, 1-(3-nitrophenyl)pyrrolidin-2-one (600 mg, 2.91 mmol, 1.0 equiv.) was added to methanol (20 mL), then Pd / C (10%) (50 mg) was added, and the mixture was stirred at room temperature under a hydrogen atmosphere for 14 hours. After monitoring the completion of the reaction by LC-MS, Pd-C was removed by suction filtration, and the reaction solution was concentrated to obtain the title compound (400 mg, yield 78%). LC-MS (ESI) [ M + H] + = 177.1。

[0428] Step 3: Preparation of 1-(3-((6-(3S,4S-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)phenyl)pyrrolidin-2-one

Chemical formula

[0429] (6-Chloropyrimidin-4-yl)((3S,4S)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone (67 mg, 0.18 mmol, 1.0 equiv) was dissolved in 1,4-dioxane (3 mL), and 1-(3-aminophenyl)pyrrolidin-2-one (38 mg, 0.22 mmol, 1.2 equiv), palladium acetate (6 mg, 0.027 mmol, 0.15 equiv), BINAP (1,1'-binaphthalene-2,2'-bis(diphenylphosphino)) (33 mg, 0.054 mol, 0.3 equiv), and cesium carbonate (146 mg, 0.45 mmol, 2.5 equiv) were added. The mixture was heated and stirred by microwave at 80 °C for 1 hour. After monitoring the completion of the reaction by LC-MS, it was cooled to room temperature, filtered, and the crude product was separated and purified using Prep-HPLC (aqueous 10 mmol / L NH4NH3, MeCN) to obtain the title compound (20 mg, purity 95.44%, yield 21.65%).

[0430] LC-MS (ESI) [M + H] + = 513.5、 1 H NMR (400 MHz, MeOD-d4) δ 9.87 (d, J = 8.0 Hz, 1H), 8.65 (d, J = 8.4 Hz, 1H), 8.03 (s, 1H), 7.58 (t, J = 5.6 Hz, 1H), 7.35 (td, J = 8.0 Hz, 2.4 Hz, 1H), 7.28~7.26 (m, 1H), 7.11~7.03 (m, 4H), 6.88 (dd, J = 6.4 Hz, 0.8 Hz, 1H), 4.85~4.74 (m, 1H), 4.52~4.33 (m, 1H), 3.87~3.81 (m, 4H), 3.78~3.75 (m, 1H), 3.69~3.62 (m, 1H), 3.31~3.30 (m, 1H), 3.07~3.01 (m, 0.5H), 2.92~2.74 (m, 5H), 2.69~2.60 (m, 1.5H), 2.54~2.53 (m, 1H), 2.11~2.04 (m, 2H), 1.88~1.74 (m, 1H), 1.58~1.50 (m, 1H).

[0431] Examples 140 - 190 The compounds of Examples 140 - 190 were synthesized using the method of Example 139, and the structures and specific identification data (LC-MS and 1 H NMR) of the compounds were as follows. [Table 3 - 1] [Table 3 - 2] [Table 3 - 3] [Table 3 - 4] [Table 3 - 5]

Table 3-6

Table 3-7

Table 3-8

Table 3-9

[0432] Example 183: Preparation of ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone (6-((1-benzoylpiperidin-4-yl)amino)-2-isopropoxypyrimidin-4-yl))

[0433] Step 1: Preparation of 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylic acid

Chem.

[0434] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (500 mg, 1.347 mmol, 1.0 equiv.) was dissolved in acetonitrile (5 mL), potassium trimethylsilyl (0.1729 g, 1.347 mmol, 1.0 equiv.) was added, and the mixture was stirred at 25 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, it was concentrated, dissolved in a 10% dichloromethane-methanol solution, filtered, and the filtrate was concentrated to obtain the title compound (480 mg, yield 99.8%). LC-MS (ESI) [M + H] + = 357.2.

[0435] Step 2: Preparation of 6 - ((1 - (tert - butoxycarbonyl)piperidin - 4 - yl)amino)-2 - isopropoxypyrimidine - 4 - carboxylic acid

Chem.

[0436] 6 - ((1 - (tert - butoxycarbonyl)piperidin - 4 - yl)amino)-2 - chloropyrimidine - 4 - carboxylic acid (480 mg, 1.35 mmol, 1.0 equiv.) was dissolved in isopropanol (5 mL), and potassium tert - butoxide (628.8 mg, 5.4 mmol, 4.0 equiv.) was added. The mixture was stirred at 90 °C for 3 hours. The completion of the reaction was monitored by LC - MS, concentrated, and purified by Prep - TLC (MeOH:DCM = 10%) to obtain the title compound (529 mg, yield 99.3%). LC - MS (ESI) [M + H] + = 381.3。

[0437] Step 3: Preparation of tert - butyl 4 - ((6 - (3R,4R - 4 - (3,4 - dihydroisoquinolin - 2(1H)-yl)-3 - hydroxypiperidine - 1 - carbonyl)-2 - isopropoxypyrimidin - 4 - yl)amino)piperidine - 1 - carboxylate

Chem.

[0438] 6 - ((1 - (tert - Butoxycarbonyl)piperidin - 4 - yl)amino)-2 - isopropoxypyrimidine - 4 - carboxylic acid (200 mg, 0.526 mmol, 1.0 equiv.) was dissolved in N,N - dimethylformamide (2 mL), and (3R,4R)-4-(3,4 - dihydroisoquinolin - 2(1H)-yl)piperidin - 3 - ol (146.6 mg, 0.631 mmol, 1.2 equiv.), HATU (299.8 mg, 0.789 mmol, 1.5 equiv.) and N,N - diisopropylethylamine (203.8 mg, 1.577 mmol, 3.0 equiv.) were added, and the mixture was stirred at room temperature for 2 hours. After monitoring the completion of the reaction by LC - MS, the mixture was extracted and concentrated, and purified by reverse phase to obtain the compound (80 mg, yield 25.6%). LC - MS (ESI) [M + H] + = 595.4。

[0439] Step 4: Preparation of ((3R,4R)-4-(3,4 - dihydroisoquinolin - 2(1H)-yl)-3 - hydroxypiperidin - 1 - yl)(2 - isopropoxy - 6-(piperidin - 4 - ylamino)pyrimidin - 4 - yl)methanone

Chemical Structure

[0440] tert - Butyl 4 - ((6-(3R,4R - 4-(3,4 - dihydroisoquinolin - 2(1H)-yl)-3 - hydroxypiperidin - 1 - carbonyl)-2 - isopropoxypyrimidin - 4 - yl)amino)piperidine - 1 - carboxylate (80 mg, 0.134 mmol, 1.0 equiv.) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (0.5 mL) was added, and the mixture was stirred at 25 °C for 1 hour. After monitoring the completion of the reaction by LC - MS, the mixture was concentrated to obtain the title compound (70 mg, crude product). LC - MS (ESI) [M + H] + = 495.3。

[0441] Step 5: Preparation of ((6 - ((1 - benzoylpiperidin - 4 - yl)amino)-2 - isopropoxypyrimidin - 4 - yl))((3R,4R)-4-(3,4 - dihydroisoquinolin - 2(1H)-yl)-3 - hydroxypiperidin - 1 - yl)methanone

Chemical Structure

[0442] ((3R,4R)-4-(3,4 - dihydroisoquinolin - 2(1H)-yl)-3 - hydroxypiperidin - 1 - yl)(2 - isopropoxy - 6-(piperidin - 4 - ylamino)pyrimidin - 4 - yl)methanone (70 mg, 0.142 mmol, 1.0 equiv.) was dissolved in dichloromethane (1 mL), N,N - diisopropylethylamine (54.77 mg, 0.425 mmol, 3.0 equiv.) was added, and the mixture was stirred for 15 minutes in an ice bath. Then, benzoyl chloride (21.96 mg, 0.156 mmol, 1.1 equiv.) was added, and the temperature was gradually raised to room temperature and stirred for 2 hours. After monitoring the completion of the reaction by LC - MS, the mixture was concentrated to obtain a crude product, which was prepared by reverse phase to give the title compound (37 mg, yield 43.7%).

[0443] LC - MS (ESI) [M + H] + = 599.3; 1 1H NMR (400 MHz, CDCl3) δ 7.49 - 7.36 (m, 5H), 7.14 (m, 3H), 7.07 - 6.95 (m, 1H), 6.21 (d, J = 22.4 Hz, 1H), 5.32 - 5.12 (m, 1H), 5.09 - 4.55 (m, 3H), 4.29 (m, 1H), 3.97 (m, 1H), 3.85 - 3.59 (m, 3H), 3.27 - 2.87 (m, 6H), 2.85 - 2.53 (m, 3H), 2.07 - 2.01 (m, 3H), 1.77 - 1.51 (m, 3H), 1.41 - 1.28 (m, 6H).

[0444] Example 184: Preparation of (2-chloro-6-(cyclobutylamino)pyrimidin-4-yl)(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone

[0445] Step 1: Preparation of 2-chloro-6-(cyclobutylamino)pyrimidine-4-carboxylic acid

Chem.

[0446] Methyl 2-chloro-6-(cyclobutylamino)pyrimidine-4-carboxylate (1.5 g, 6.207 mmol, 1.0 equiv.) was added to water (5 mL) and THF (20 mL) at room temperature, lithium hydroxide (297.3 mg, 12.414 mmol, 2.0 equiv.) was added, and the mixture was stirred at room temperature for 1 hour. After monitoring the completion of the reaction by LC-MS, the reaction solution was concentrated to remove the organic solvent, adjusted to pH = 3 with 1 M hydrochloric acid, and the reaction system was extracted three times with dichloromethane:methanol (V / V = 10 / 1) (30 mL × 5). The organic phases were combined, dried, filtered, and concentrated to obtain the title compound (1.5 g, crude product). LC-MS (ESI) [M + H] + = 442.4.

[0447] Step 2: Preparation of (2-chloro-6-(cyclobutylamino)pyrimidin-4-yl)(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone

Chem.

[0448] Using the method of Step 2 of Example 5, the title compound (1.14 g, yield: 39.7%) was synthesized. LC-MS (ESI) [M+H] + = 442.3; 11H NMR (400 MHz, DMSO-d6) δ 8.58 - 8.35 (m, 1H), 7.10 - 6.88 (m, 4H), 6.52 - 6.45 (m, 1H), 4.83 - 4.73 (m, 1H), 4.42 - 4.03 (m, 2H), 3.86 - 3.76 (m, 2H), 3.70 - 3.53 (m, 2H), 3.05 - 2.73 (m, 5H), 2.69 - 2.57 (m, 1H), 2.33 - 2.28 (m, 2H), 2.03 - 1.82 (m, 3H), 1.73 - 1.61 (m, 2H), 1.56 - 1.40 (m, 1H), 1.23 - 1.21 (m, 1H).

[0449] Example 185: Preparation of trans-(6-(Cyclobutylamino)-2-(2-(dimethylamino)ethoxy)pyrimidin-4-yl)(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone

Chemical Structure

[0450] The compound of Example 184 (0.1 g, 0.226 mmol, 1.0 equiv.) was added to n-butanol (2 mL), then 2-(dimethylamino)ethan-1-ol (201.7 mg, 2.263 mmol, 1.0 equiv.) was added, and the mixture was stirred at 140 °C for 1 hour under microwave irradiation. The completion of the reaction was monitored by LC-MS, and purification was performed by reverse-phase preparative HPLC separation (C18, 10 mmol / L aqueous NH4HCO3 solution, MeCN), followed by lyophilization to obtain the title compound (27.36 mg, yield 24.4%).

[0451] LC-MS (ESI) [M+H] + = 495.4; 11H NMR (400 MHz, DMSO-d6) δ 7.93-7.90 (m, 1H), 7.10-7.02 (m, 4H), 6.13 (d, J = 5.2 Hz, 1H), 4.81-4.71 (m, 1H), 4.46-4.24 (m, 4H), 3.95-3.86 (m, 4H), 3.01-2.86 (m, 1H), 2.84-2.75 (m, 4H), 2.67-2.53 (m, 4H), 2.36-2.25 (m, 2H), 2.20 (s, 3H), 2.18 (s, 3H), 1.91-1.63 (m, 5H), 1.52-1.43 (m, 1H).

[0452] Example 186: Preparation of trans-(6-(cyclobutylamino)-2-phenoxypyrimidin-4-yl)(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone

Chemical formula

[0453] The compound of Example 184 (0.1 g, 0.226 mmol, 1.0 equiv.) was added to n-butanol (2 mL), potassium phenoxide (89.7 mg, 0.679 mmol, 3.0 equiv.) was added, and the mixture was stirred at 140 °C for 1 hour by microwave. The completion of the reaction was monitored by LC-MS, and the title compound (4.21 mg, yield 3.7%) was obtained by reverse-phase preparative HPLC separation (C18, 10 mmol / L aqueous NH4HCO3 solution, MeCN) and lyophilization.

[0454] LC-MS (ESI) [M+H] + = 500.5; 11H NMR (400 MHz, CD3OD) δ 7.44 - 7.37 (m, 2H), 7.25 - 7.10 (m, 6H), 7.05 - 7.01 (m, 1H), 6.27 - 6.24 (m, 1H), 4.70 - 4.49 (m, 1H), 4.29 - 4.23 (m, 1H), 3.91 - 3.77 (m, 3H), 3.71 - 3.61 (m, 1H), 3.02 - 2.59 (m, 8H), 2.23 - 1.71 (m, 7H).

[0455] Example 187: Preparation of 1-(4-((6-(3R,4R-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-isobutoxypyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0456] Step 1: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate

Chemical Structure

[0457] Methyl 2,6-dichloropyrimidine-4-carboxylate (6 g, 28.98 mmol), 1-acetylpiperidin-4-amine hydrochloride (5.7 g, 31.88 mmol) and DIPEA (14.98 g, 115.94 mmol) were dissolved in acetonitrile (50 mL). The mixture was stirred at room temperature (25 - 30 °C) for 2 hours. It was extracted three times with 100 mL of ethyl acetate each time. The ethyl acetate phases were combined, washed once with 50 mL of water, once with 50 mL of saturated brine, dried over anhydrous sodium sulfate for 10 minutes, filtered, and the crude product was separated and purified by column chromatography using DCM:MeOH = 100:1 to obtain the product (5.8 g, yield 64%). LC-MS (ESI) [M + H] + = 313.2.

[0458] Step 2: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-isobutoxypyrimidine-4-carboxylic acid [Chemical formula]

[0459] At 20 °C, methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (250 mg, 0.80 mmol, 1.0 equiv.) was dissolved in anhydrous dioxane (3 mL), 2-methylpropan-1-ol (178 mg, 2.4 mmol, 1.1 equiv.) and potassium tert-butoxide (180 mg, 1.6 mmol, 2 equiv.) were added, and the mixture was stirred at 80 °C for 2 hours. After monitoring the completion of the reaction by LC-MS, the product was concentrated, and the crude product was purified by preparative HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) and lyophilized to obtain the title compound (35 mg, yield: 13%). LC-MS (ESI) [M + H] + = 337.2

[0460] Step 3: Preparation of 1-(4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-isobutoxypyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one [Chemical formula]

[0461] At 20 °C, the carboxylic acid intermediate (35 mg, 0.1 mmol, 1.0 equiv.) was added to DMF (2 mL), then 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (28 mg, 0.12 mmol, 1.2 equiv.), HOAt (N-hydroxy-7-azabenzotriazole) (16 mg, 0.12 mmol, 2 equiv.), and 3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (23 mg, 0.22 mmol, 1.0 equiv.) were added, and the mixture was stirred at room temperature for 2 hours at 20 °C. After monitoring the completion of the reaction by LC-MS, the crude product was separated and purified by preparative HPLC (C18, in 10 mmol / L aqueous NH4HCO3 / acetonitrile), and freeze-dried to obtain the title compound (15 mg, yield 27.3%).

[0462] LC-MS (ESI) [M+H] + = 551.4; 1 1H NMR (400 MHz, CD3OD) δ 7.13 - 7.05 (m, 4H), 6.21 (d, J = 7.6 Hz, 1H), 4.72 - 4.55 (m, 1H), 4.43 (d, J = 13.2 Hz, 1H), 4.20 - 4.11 (m, 3H), 4.01 - 3.79 (m, 5H), 3.28 - 3.06 (m, 2H), 3.00 - 2.87 (m, 5H), 2.82 - 2.68 (m, 2H), 2.13 - 2.05 (m, 5H), 2.02 - 1.89 (m, 2H), 1.73 - 1.67 (m, 1H), 1.53 - 1.43 (m, 2H), 1.04 (t, J = 6.4 Hz, 6H).

[0463] Example 188: Preparation of 1-(4-((6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(pentyl-3-yloxy)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0464] Step 1: 6 - ((1 - Acetylpiperidin - 4 - yl)amino) - 2 - (pentan - 3 - yloxy)pyrimidine - 4 - carboxylic acid

Chem.

[0465] Methyl 6 - ((1 - acetylpiperidin - 4 - yl)amino) - 2 - chloropyrimidine - 4 - carboxylate (300 mg, 0.96 mmol, 1.0 equiv.), 3 - pentanol (845 mg, 9.6 mmol, 10 equiv.) were dissolved in anhydrous t - butanol (5 mL), sodium t - butoxide (369 mg, 3.84 mmol, 4.0 equiv.) was added, and the mixture was reacted at 100 °C for 16 h. After monitoring the completion of the reaction by LC - MS, the reaction solution was adjusted to pH 4 with 1 M hydrochloric acid, and the solvent was dried to obtain the crude product (700 mg). LC - MS (ESI) [M + H] + = 351.2.

[0466] Step 2: Preparation of 1 - (4 - ((6 - (3R,4R - 4 - (3,4 - dihydroisoquinolin - 2(1H) - yl) - 3 - hydroxypiperidine - 1 - carbonyl) - 2 - (pentan - 3 - yloxy)pyrimidin - 4 - yl)amino)piperidin - 1 - yl)ethan - 1 - one

Chem.

[0467] 6-((1-Acetylpiperidin-4-yl)amino)-2-(pentan-3-yloxy)pyrimidine-4-carboxylic acid (90 mg, 0.26 mmol, 1.0 equiv.), (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (60 mg, 0.26 mmol, 1.0 equiv.), EDCI (74 mg, 0.38 mmol, 1.5 equiv.) and HOAt (52 mg, 0.38 mmol, 1.5 equiv.) were dissolved in DMF (N,N-dimethylformamide) (2 mL) and reacted at 25 °C for 1 h. After monitoring the completion of the reaction by LC-MS, the mixture was concentrated and the crude product was separated and purified by reverse-phase HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to give the title compound (26.33 mg, yield: 18%).

[0468] LC-MS (ESI) [ M + H] + = 565.4, 1 H NMR (400 MHz, CDCl3) δ 7.17 - 7.10 (m, 3H), 7.03 - 7.01 (m, 1H), 6.22 - 6.17 (m, 1H), 5.29 - 5.17 (m, 1H), 5.00 - 4.90 (m, 1H), 4.77 - 4.74 (m, 1H), 4.51 - 4.47 (m, 1H), 4.24 - 4.12 (m, 1H), 3.96 - 3.92 (m, 1H), 3.81 - 3.78 (m, 1H), 3.72 - 3.62 (m, 2H), 3.24 - 3.21 (m, 1H), 3.06 - 2.64 (m, 9H), 2.11 - 2.10 (m, 4H), 2.02 - 1.94 (m, 2H), 1.70 - 1.60 (m, 5H), 1.41 - 1.38 (m, 2H), 0.96 - 0.89 (m, 6H).

[0469] Example 189: Preparation of (6-((1-(cyclobutanecarbonyl)piperidin-4-yl)amino)-2-isopropoxypyrimidin-4-yl)((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone [Chemistry]

[0470] ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-isopropoxy-6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone (70 mg, 0.142 mmol, 1.0 equiv.) was dissolved in dichloromethane (1 mL), N,N-diisopropylethylamine (54.77 mg, 0.425 mmol, 3.0 equiv.) was added, and the mixture was stirred in an ice bath for 15 minutes. Then, cyclobutanecarbonyl chloride (21.96 mg, 0.156 mmol, 1.1 equiv.) was added, and the temperature was gradually raised to room temperature and stirred for 2 hours. After monitoring the completion of the reaction by LC-MS, the mixture was concentrated to obtain a crude product, which was prepared by reverse phase to obtain the title compound (54 mg, yield 66.2%).

[0471] LC-MS (ESI) [M+H] + = 577.4; 1 1H NMR (400 MHz, CDCl3) δ 7.17 - 7.11 (m, 3H), 7.04 - 7.02 (m, 1H), 6.23 - 6.17 (m, 1H), 5.22 - 5.17 (m, 1H), 5.02 - 4.74 (m, 2H), 4.52 - 4.49 (m, 1H), 4.30 - 4.27 (m, 1H), 4.00 - 3.96 (m, 1H), 3.76 - 3.68 (m, 3H), 3.31 - 3.20 (m, 1H), 3.19 - 2.78 (m, 5H), 2.75 - 2.53 (m, 4H), 2.37 - 2.33 (m, 2H), 2.16 - 1.95 (m, 7H), 1.66 - 1.65 (m, 1H), 1.38 - 1.33 (m, 8H).

[0472] Examples 190 - 294 Using the methods of Examples 183, 185 and 186, the compounds of Examples 190 - 294 were synthesized, and the structures of the compounds and specific identification data (LC-MS and 1The 1H NMR was as follows. [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5] [Table 4-6] [Table 4-7] [Table 4-8] [Table 4-9] [Table 4-10] [Table 4-11] [Table 4-12] [Table 4-13] [Table 4-14] [Table 4-15] [Table 4-16] [Table 4-17]

Table 4-18

Table 4-19

Table 4-20

Table 4-21

[0473] Example 295: Preparation of 1-(4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-isopropoxypyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0474] Step 1: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-isopropoxypyrimidine-4-carboxylic acid

Chemical formula

[0475] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (3.25 g, purity 50% (including 50% DIPEA hydrochloride), 5.196 mmol, 1.0 equiv.) and sodium tert-butoxide (4.0 g, 41.62 mmol, 8.01 equiv.) were weighed in a one-neck flask, and anhydrous isopropanol (104 mL) was added. Under a nitrogen atmosphere, the reaction was carried out at 100 °C for 2 hours. The completion of the reaction was monitored by LC-MS, the pH was adjusted to 3-4, filtered, and the filtrate was dried with a rotary evaporator with a solvent to obtain a crude product (110 mg). The crude product was purified by reverse-phase HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain the title compound (755 mg, yield: 44.6%). LC-MS (ESI) [M + H] + = 323.3。

[0476] Step 2: Preparation of 1-(4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-carbonyl)-2-isopropoxypyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chemical formula

[0477] 6-((1-Acetylpiperidin-4-yl)amino)-2-isopropoxypyrimidine-4-carboxylic acid (550 mg, 1.706 mmol, 1 equiv.), EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) (495 mg, 2.582 mmol, 1.51 equiv.) and HOAt (N-hydroxy-7-azabenzotriazole) (352 mg, 2.586 mmol, 1.52 equiv.) were dissolved in DMF (5.5 mL), stirred for 5 minutes, and then a DMF solution (3.5 mL) of (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (396 mg, 1.534 mmol, 0.9 equiv.) was added. The reaction mixture was stirred at room temperature for 1 hour. The solvent was dried using a rotary evaporator, and the crude product was purified by reverse-phase HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain the target compound (536 mg, yield 58.6%).

[0478] LC-MS (ESI) [M+H] + = 537.3; 11H NMR (400 MHz, CDCl3) δ 7.19 - 7.08 (m, 3H), 7.07 - 6.99 (m, 1H), 6.28 - 6.14 (m, 1H), 5.69 - 5.28 (m, 1H), 5.26 - 5.14 (m, 1H), 5.03 - 4.94 (m, 0.37H), 4.80 - 4.70 (m, 0.64H), 4.55 - 4.44 (m, 1H), 4.30 - 4.13 (m, 1H), 3.99 - 3.91 (m, 1H), 3.90 - 3.61 (m, 4H), 3.26 - 3.15 (m, 1H), 3.10 - 2.56 (m, 8H), 2.14 - 1.88 (m, 6H), 1.72 - 1.58 (m, 1H), 1.46 - 1.30 (m, 8H).

[0479] Example 296: Preparation of 1-(4-((6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(pyrrolidin-1-yl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0480] Step 1: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate

Chemical Structure

[0481] Methyl 2,6-dichloropyrimidine-4-carboxylate (6 g, 28.98 mmol), 1-acetylpiperidin-4-amine hydrochloride (5.7 g, 31.88 mmol), and DIPEA (14.98 g, 115.94 mmol) were dissolved in acetonitrile (50 ml). The mixture was stirred at room temperature (25 - 30 °C) for 2 hours. It was extracted three times with 100 mL of ethyl acetate each time. The ethyl acetate phases were combined, washed once with 50 mL of water, once with 50 mL of saturated brine, dried over anhydrous sodium sulfate for 10 minutes, filtered, and the crude product was separated and purified by column chromatography using DCM:MeOH = 100:1 to obtain the product (5.8 g, yield 64%). LC-MS (ESI) [ M + H] + = 313.2。

[0482] Step 2: Preparation of 6 - ((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylic acid hydrochloride

Chemical formula

[0483] Methyl 6 - ((1-acetylpiperidin-4-yl)amino)-2-methylpyrimidine-4-carboxylate (5 g, 16.02 mmol) was dissolved in THF (36 mL) and methanol (36 mL), and then an aqueous solution of lithium hydroxide (16 mL, 32 mmol, 2M) was added. The reaction mixture was stirred at room temperature (25 - 30 °C) for 2 hours. The completion of the reaction was monitored by TLC. 1M hydrochloric acid (molar concentration) was added to adjust the pH to 6 - 7, and it was directly dried on a rotary evaporator to obtain the crude product (5.8 g). LC-MS (ESI) [ M + H] + = 299.0。

[0484] Step 3: Preparation of 1-(4-((2-chloro-6-((trans)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chemical Structure

[0485] 6-((1-Acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylic acid hydrochloride (500 mg, 1.67 mmol), HATU (636.4 mg, 1.67 mmol), trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (388.8 mg, 1.67 mmol) and DIPEA (1.08 g, 8.37 mmol) were dissolved in DMF (6.5 mL), and the reaction mixture was stirred at room temperature (25 - 30 °C) for 2 hours. LC-MS was used to monitor the completion of the reaction. The mixture was extracted three times with 20 mL portions of ethyl acetate. The combined ethyl acetate phases were washed once with 10 mL of water, once with 10 mL of saturated brine, dried over anhydrous sodium sulfate for 10 minutes, filtered, and the crude product was separated and purified by column chromatography on DCM:MeOH = 20:1 to obtain the title compound (200 mg). LC-MS (ESI) [M + H] + = 513.2

[0486] Step 4: Preparation of 1-(4-((6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(pyrrolidin-1-yl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chemical Structure

[0487] 1-(4-((2-Chloro-6-((trans)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one (133 mg, 0.195 mmol) and tetrahydropyrrole (55 mg, 0.78 mmol) were dissolved in 1,4-dioxane (2 mL), Et3N (0.3 mL) was added, and the mixture was stirred at 100 °C for 2 h while LC-MS monitored the completion of the reaction. EA extraction (3 * 10 mL), the EA phases were combined, washed once with 10 mL of water and 10 mL of saturated brine respectively, dried over anhydrous sodium sulfate, filtered, and purified by preparative reverse-phase HPLC ([ 18 18 , 10 mmol / L aqueous C 43 / acetonitrile), to obtain the title compound (77.47 mg, 72% yield).

[0488] LC-MS (ESI) [M+H] + = 548.3; 1 H NMR (400 MHz, CDCl3) δ 7.18 - 7.10 (m, 3H), 7.04 - 7.02 (m, 1H), 5.89 - 5.86 (m, 1H), 5.03 (m, 0.3H), 4.67 (s, 1H), 4.60 - 4.44 (m, 1.7H), 4.42 - 4.27 (m, 1H), 4.19 (s, 1H), 4.04 - 3.64 (m, 5H), 3.52 (s, 4H), 3.19 (t, J = 12.6 Hz, 1H), 3.12 - 2.88 (m, 5H), 2.88 - 2.51 (m, 3H), 2.19 - 2.08 (m, 4H), 2.08 - 1.90 (m, 6H), 1.90 - 1.73 (m, 1H), 1.48 - 1.31 (m, 2H).

[0489] Example 297: Preparation of 1-(4-((6-((trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(trifluoromethyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0490] Step 1: Preparation of 1-(4-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chem.

[0491] 4,6-Dichloro-2-(trifluoromethyl)pyrimidine (1.09 g, 5.02 mmol), 1-acetylpiperidin-4-amine hydrochloride (0.89 g, 5.02 mmol) and DIPEA (1.95 g, 15.07 mmol) were dissolved in acetonitrile (10 mL) and stirred at room temperature for 2 hours. The reaction was monitored by LC-MS until completion, followed by extraction with EA (3 * 20 mL), mixing of the EA phases, washing once with 20 mL of water and once with 20 mL of saturated brine, drying over anhydrous sodium sulfate, filtration, and separation and purification of the crude product by column chromatography using DCM:MeOH = 50:1 to obtain the title compound (1.174 g, yield 54%). LC-MS (ESI) [M + H] + = 323.1。

[0492] Step 2: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethyl)pyrimidine-4-carboxylate

Chem.

[0493] 1-(4-((6-Chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one (0.3 g, 0.93 mmol), Et3N (282 mg, 2.79 mmol) and Pd(dppf)Cl2 (68 mg, 0.093 mmol) were dissolved in a mixed solution of methanol (15 mL) and DMF (15 mL), and the reaction mixture was heated at 100 °C for 30 h in a CO atmosphere at 2 atm. Monitored by LC-MS, a part of the reactant was converted to the product. The solvent was dried by a rotary evaporator, extracted with EA (3 * 20 mL), the EA phases were combined, washed once with 20 mL of water and once with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the crude product was separated and purified by column chromatography using DCM:MeOH = 50:1 to obtain the title compound (100 mg, yield 31%). LC-MS (ESI) [ M + H] + = 347.2。

[0494] Step 3: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethyl)pyrimidine-4-carboxylic acid hydrochloride

Chemical formula

[0495] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethyl)pyrimidine-4-carboxylate (100 mg, 0.289 mmol) was dissolved in THF (4 mL), and then an aqueous lithium hydroxide solution (0.43 mL, 0.87 mmol, 2 M) was added. Stirred at room temperature for 1 h, and LC-MS monitored the completion of the reaction. Extracted with EA (3 × 10 mL), the aqueous phase was retained, 1 M hydrochloric acid was added to adjust the pH to 6 - 7, and dried directly by a rotary evaporator to obtain the crude product (120 mg). LC-MS (ESI) [ M + H] + = 333.2。

[0496] Step 4: Preparation of 1-(4-((6-((trans-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(trifluoromethyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chemical Structure

[0497] According to the method of Step 2 in Example 5, a condensation reaction was carried out using 6-((1-acetylpiperidin-4-yl)amino)-2-(trifluoromethyl)pyrimidine-4-carboxylic acid hydrochloride (100 mg, 0.27 mmol, 1.0 equiv.) and trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (66 mg, 0.28 mmol, 1.04 equiv.). The crude product was separated and purified by reverse-phase HPLC (C18, 10 mmol / L aqueous solution / acetonitrile) to obtain the title compound (104.56 mg, yield 70%).

[0498] LC-MS (ESI) [M+H] + = 547.2; 1 1H NMR (400 MHz, CDCl3) δ 7.19-7.11 (m, 3H), 7.05-7.03 (m, 1H), 6.82-6.72 (m, 1H), 5.90-5.66 (m, 1H), 5.00-4.71 (m, 1H), 4.57 (d, J = 13.0 Hz, 1H), 4.35-3.94 (m, 3H), 3.90-3.65 (m, 4H), 3.24 (t, J = 13.2 Hz, 1H), 3.16-3.00 (m, 2H), 3.00-2.92 (m, 2H), 2.92-2.59 (m, 4H), 2.12 (d, J = 2.4 Hz, 4H), 2.08-1.97 (m, 2H), 1.94-1.76 (m, 1H), 1.53-1.31 (m, 2H).

[0499] Example 298: Preparation of trans-1-(4-((6-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(1H-imidazol-1-yl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0500] Step 1: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-(1H-imidazol-1-yl)pyrimidine-4-carboxylic acid

Chemical formula

[0501] 6-((1-Acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylic acid (400 mg, 1.339 mmol, 1 equiv.) and imidazole (182 mg, 2.678 mmol, 2 equiv.) were dissolved in DMA (20 ml), then cesium carbonate (1.31 g, 4.017 mmol, 3 equiv.) was added, and the mixture was stirred at 100 °C for 4 hours. The completion of the reaction was monitored by LC-MS, 10 mL of water was added to the reaction solution to quench it, and it was extracted several times with DCM. The organic phase was dried and concentrated to obtain the title compound (3.0 g, crude product). LC-MS (ESI) [M + H] + = 331.2.

[0502] Step 2: Preparation of trans-1-(4-((6-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(1H-imidazol-1-yl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chemical formula

[0503] According to the method of the second step of Example 5, 6-((1-acetylpiperidin-4-yl)amino)-2-(1H-imidazol-1-yl)pyrimidine-4-carboxylic acid (crude product 420 mg, 1.272 mmol, 1 equiv.) and trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (325 mg, 1.399 mmol, 1.1 equiv.) were used to carry out a condensation reaction, and the crude product was prepared by reverse-phase HPLC ( 43 aqueous solution / acetonitrile) to obtain the title compound (75.42 mg, yield 10.9%).

[0504] LC-MS (ESI) [M+H] + = 545.3; 1 H NMR (400 MHz, DMSO-d6) δ 8.53-8.40 (m, 1H), 8.09 (t, J = 8.1 Hz, 1H), 7.87-7.80 (m, 1H), 7.17-7.03 (m, 5H), 6.74-8.47 (m, 1H),4.84-4.74 (m, 1H), 4.56-4.14 (m, 3H), 3.84-3.79 (m, 4H), 3.69-3.65 (m, 1H), 3.08-3.02 (m, 1H), 2.97-2.76 (m, 6H), 2.71-2.57 (m, 2H), 2.02 (s, 3H), 1.94-1.79 (m, 2H), 1.79-1.48 (m, 2H), 1.43-1.29 (m, 2H).

[0505] Example 299: Preparation of 1-(4-((6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-morpholinopyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0506] First step: Preparation of methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate

Chemical formula

[0507] Methyl 2,6-dichloropyrimidine-4-carboxylate (2 g, 9.66 mmol), 1-acetylpiperidin-4-amine hydrochloride (1.9 g, 10.63 mmol) and N,N-diisopropylethylamine (4.99 g, 38.65 mmol) were dissolved in acetonitrile (50 ml). The mixture was stirred at 25 °C for 2 hours. Ethyl acetate (50 mL) was used for extraction three times. The ethyl acetate phases were combined, washed once with 20 mL of water, washed once with 20 mL of saturated brine, dried over anhydrous sodium sulfate for 10 minutes, filtered, and the crude product was separated and purified by column chromatography using DCM:MeOH = 100:1 to obtain methyl 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (2.8 g, yield 92%). LC-MS (ESI) [ M + H] + = 313.2。

[0508] Step 2: Preparation of 6-((1-acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylic acid

Chemical formula

[0509] Methyl 6-((1-acetylpiperidin-4-yl)amino)-2-methylpyrimidine-4-carboxylate (200 mg, 0.639 mmol) was dissolved in THF (4 mL), and then 2M lithium hydroxide aqueous solution (0.64 mL, 1.28 mmol, 2 equiv.) was added. The reaction solution was stirred at room temperature (25 °C) for 2 hours. The completion of the reaction was monitored by TLC. 1M hydrochloric acid was added to adjust the pH to 6 - 7, and the solution was directly dried by a rotary evaporator to obtain the crude product (400 mg). LC-MS (ESI) [ M + H] + = 299.2。

[0510] Step 3: Preparation of 1-(4-((2-((3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)oxy)-6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chem.

[0511] 6-((1-Acetylpiperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylic acid (380 mg, 1.27 mmol), HATU (2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (725.5 mg, 1.91 mmol), trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (265 mg, 1.14 mmol) and DIPEA (N,N-diisopropylethylamine) (0.822 g, 6.36 mmol) were dissolved in DMF (N,N-dimethylformamide) (5 mL), and the reaction mixture was stirred at room temperature for 2 hours at 25 °C. The completion of the reaction was monitored by LC-MS, and the mixture was extracted three times with 20 mL of ethyl acetate each time. The combined ethyl acetate phases were washed once with 10 mL of water, once with 10 mL of saturated brine, dried over anhydrous sodium sulfate for 10 minutes, filtered, and the crude product was separated and purified by column chromatography using DCM:MeOH = 20:1 to obtain the title compound (200 mg). LC-MS (ESI) [M + H] + = 613.3。

[0512] Step 4: Preparation of 1-(4-((6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-morpholinopyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chem.

[0513] 1-(4-((2-((3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)oxy)-6-(trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one (40 mg, 0.64 mmol) was dissolved in 1,4-dioxane (1.5 mL), and Et3N (19.2 mg, 0.192 mmol) and morpholine (16 mg, 0.192 mmol) were added. The mixture was stirred at 100 °C for 1 h, and LC-MS monitored the completion of the reaction. It was extracted three times with 10 mL portions of EA, and the combined ethyl acetate phase was washed once with 10 mL of water and once with 10 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the crude product was prepared by reverse-phase HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to give the title compound (25.59 mg, yield 69%).

[0514] LC-MS (ESI) [M + H] + = 564.4, 1 H NMR (400 MHz, DMSO-d6) δ 7.25 (s, 1H), 7.13~6.99 (m, 4H), 5.86 (s, 1H), 4.83~4.63 (m, 1H), 4.52~4.28 (m, 1H), 4.19 (d, J = 12.6 Hz, 1H), 4.02 (s, 1H), 3.89~3.70 (m, 4H), 3.70~3.51 (m, 9H), 3.17 (t, J = 12.4 Hz, 1H), 3.02~2.87 (m, 1.5H), 2.87~2.69 (m, 5H), 2.69~2.53 (m, 1.5H), 2.05~1.97 (m, 3H), 1.97~1.69 (m, 3H), 1.56~1.44 (m, 1H), 1.44~1.32 (m, 1H), 1.32~1.18 (m, 1H).

[0515] Example 300: Preparation of (trans-1-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-morpholinyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)methanone

[0516] Step 1: Preparation of methyl 2-chloro-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylate

Chem.

[0517] Methyl 2,6-dichloropyrimidine-4-carboxylate (500 mg, 2.414 mmol, 1 equiv.) and tetrahydro-2H-pyran-4-amine (240 mg, 2.373 mmol, 3 equiv.) were dissolved in acetonitrile (12 mL), and DIPEA (920 mg, 7.116 mmol, 3 equiv.) was added. The mixture was stirred at 25 °C for 2 h. The completion of the reaction was monitored by LC-MS. The reaction solution was quenched by adding water, extracted with EA, the organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated to obtain the crude product, and separated by flash chromatography (silica gel, PE:EA = 1:10) to obtain the title compound (720 mg, crude). LC-MS (ESI) [ M + H] + = 272.1.

[0518] Step 1: Preparation of methyl 2-morpholino-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylate

Chem.

[0519] Methyl 2-chloro-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylate (720 mg, 2.582 mmol, 1 equiv.) and morpholine (247 mg, 2.840 mmol, 1.1 equiv.) were dissolved in acetonitrile (12 mL), DIPEA (1 g, 7.746 mmol, 3 equiv.) was added, and the mixture was stirred at 100 °C for 6 hours. The completion of the reaction was monitored by LC-MS. The reaction solution was quenched by adding water, extracted with EA, the organic layers were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and the title compound (900 g, crude) was obtained. LC-MS (ESI) [ M + H] + = 323.3.

[0520] Step 3: Preparation of 2-morpholino-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylic acid

Chemical Structure

[0521] Methyl 2-morpholinyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylate (900 mg, 2.792 mmol, 1 equiv.) was dissolved in acetonitrile (30 mL), and TMSOK (potassium trimethylsilanolate) (394 mg, 3.071 mmol, 1.1 equiv.) was added. The mixture was stirred at room temperature for 1 hour. The completion of the reaction was monitored by TLC. The reaction solution was filtered to recover the cake, water was added to the cake to dissolve it, 1 M hydrochloric acid was added to adjust the pH to 5 - 6, the solution was dried on a rotary evaporator, then dichloromethane and methanol were added to dissolve it, filtered to remove solid impurities, and the filtrate was concentrated to obtain the title compound (1 g, crude). LC-MS (ESI) [ M + H] + = 309.2.

[0522] Step 4: Preparation of 4-(trans-1-(4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-morpholinyl-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidin-4-yl)methanone

Chem.

[0523] Using the method of the second step of Example 5, 2-morpholino-6-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-4-carboxylic acid (200 mg, 0.649 mmol, 1 equiv.) and trans-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (180 mg, 0.778 mmol, 1.2 equiv.) were subjected to a condensation reaction, and the crude product was separated and purified by reverse-phase HPLC ( 43 aqueous solution / acetonitrile) to obtain the title compound (106.9 mg, yield: 31.5%).

[0524] LC-MS (ESI) [M+H] + = 523.6; 1 1H NMR (400 MHz, DMSO-d6) δ 7.25 (s, 1H), 7.11 - 7.01 (m, 4H), 5.86 (s, 1H), 4.74 (dd, J = 36.0, 3.8 Hz, 1H), 4.51 - 4.25 (m, 1H), 3.98 (s, 1H), 3.91 - 3.72 (m, 5H), 3.67 - 3.52 (m, 9H), 3.46 - 3.35 (m, 2H), 3.03 - 2.55 (m, 7H), 1.92 - 1.69 (m, 3H), 1.61 - 1.34 (m, 3H).

[0525] Preparation of Example 301: 1-(4-((6-(3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(pyrido[3,4-d]pyrimidin-7(8H)-yl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0526] Using the same method as in Example 297, the compound of Example 301 was synthesized (2.0 mg, yield: 1.2%). LC-MS (ESI) [M + H] + = 610.4.

[0527] Examples 302 to 351 Using the same method as in Example 297, the compounds of Examples 302 to 351 were synthesized, and the structures of the compounds and specific identification data (LC-MS and 1 H NMR) were as follows.

Table 5-1

Table 5-2

Table 5-3

Table 5-4

Table 5-5

Table 5-6

Table 5-7

Table 5-8

Table 5-9

Table 5-10

[0528] Examples 352 - 374 Using the method of Example 297, the compounds of Examples 352 - 374 were synthesized, and the structures and specific identification data (LC - MS and 1 H NMR) were as follows: [Table 6 - 1] [Table 6 - 2] [Table 6 - 3]

[0529] Example 375: Preparation of methyl 4 - ((1 - acetylpiperidin - 4 - yl)amino) - 6 - (3R,4R - 4 - (3,4 - dihydroisoquinolin - 2(1H) - yl) - 3 - hydroxypiperidine - 1 - carbonyl)pyrimidine - 2 - carboxylate [Chemical formula]

[0530] A solution of 1-(4-((2-chloro-6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one (700 mg, 1.36 mmol, 1 equiv.) in methanol (15 mL, 100%) was added with palladium 1,1-bis(diphenylphosphino)ferrocenium chloride (104.4 mg, 0.14 mmol, 0.1 equiv.), replaced with carbon monoxide gas three times, reacted at 75 °C for 12 hours. After monitoring the completion of the reaction by LC-MS, it was filtered, the filter cake was washed twice with DCM:MeOH = 10:1 (20 mL), the filtrate was dried with a rotary evaporator, and the crude product was passed through column chromatography (DCM:MeOH = 10:1) to obtain a crude product of the title compound (500 mg). A portion of the product (400 mg) was prepared using reverse-phase HPLC (C18, 10 mmol / L FA aqueous solution / acetonitrile) to obtain the title compound (202.2 mg).

[0531] LC-MS (ESI) [M+H] + = 537.3; 1 H NMR (400 MHz, CD3OD) δ 8.42 (s, 1H), 7.25 - 7.11 (m, 4H), 6.68 (d, J = 5.4 Hz, 1H), 4.80 - 4.56 (m, 2H), 4.48 - 4.32 (m, 2H), 4.31 - 4.18 (m, 2H), 4.06 - 3.97 (m, 1H), 3.95 (d, J = 9.2 Hz, 3H), 3.93 - 3.77 (m, 2H), 3.43 - 3.33 (m, 1H), 3.27 - 3.12 (m, 2H), 3.11 - 2.70 (m, 5H), 2.16 (s, 1H), 2.12 (d, J = 1.2 Hz, 3H), 2.10 - 1.94 (m, 2H), 1.94 - 1.77 (m, 1H), 1.59 - 1.35 (m, 2H).

[0532] Example 376: Preparation of (3S,4S)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-1-(phenylsulfonyl)piperidin-3-ol [Chemical formula]

[0533] (3S,4S)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (168 mg, 0.724 mmol, 1.0 equiv.) was dissolved in dichloromethane (10 ml), and triethylamine (83 mg, 0.826 mmol, 1.2 equiv.) was added. The ice-salt bath was cooled to about -10 °C, and then a solution of benzenesulfonyl chloride (CAS: 98-09-9) (128 mg, 0.724 mmol, 1.0 equiv.) in dichloromethane (2 ml) was slowly added to the reaction solution. After the addition, the temperature was raised to room temperature (20 °C) and stirred for half an hour, and the disappearance of the starting material was monitored by LC-MS. Water was added to the reaction solution to quench it, and it was extracted with methylene chloride. The organic layers were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated to obtain a crude product, and separated and purified by reverse-phase HPLC (C18, NH4HCO3 aqueous solution / acetonitrile) to obtain the title compound (48.5 mg, yield 18.03%).

[0534] LC-MS (ESI) [M+H] + = 373.2; 1 1H NMR (400 MHz, (CD3)2SO) δ 7.82 - 7.71 (m, 3H), 7.67 (t, J = 7.3 Hz, 2H), 7.10 - 7.02 (m, 3H), 7.01 - 6.96 (m, 1H), 4.80 (d, J = 3.9 Hz, 1H), 3.80 - 3.57 (m, 5H), 2.89 - 2.79 (m, 1H), 2.78 - 2.65 (m, 3H), 2.40 - 2.25 (m, 2H), 2.09 - 1.98 (m, 1H), 1.82 - 1.71 (m, 1H), 1.65 - 1.49 (m, 1H).

[0535] Example 377: Preparation of (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-1-(benzenesulfonyl)piperidin-3-ol [Chemical formula]

[0536] (3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (200 mg, 0.861 mmol, 1.0 equiv.) was dissolved in dichloromethane (5 mL), and triethylamine (105 mg, 1.033 mmol, 1.2 equiv.) was added. The ice-salt bath was cooled to about -10 °C, and then a solution of benzenesulfonyl chloride (160 mg, 0.904 mmol, 1.05 equiv.) in dichloromethane (5 mL) was slowly added to the reaction solution. After the addition, the temperature was raised to room temperature (20 °C) and stirred for half an hour, and the disappearance of the raw material was monitored by LC-MS. Water was added to the reaction solution to quench the reaction, and the mixture was extracted with dichloromethane. The combined organic layers were washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product, which was separated and purified by reverse-phase HPLC (C18, aqueous NH4HCO3 / acetonitrile) to obtain the title compound (49.83 mg, yield 15.6%).

[0537] LC-MS (ESI) [M+H] + = 373.2; 1 1H NMR (400 MHz, (CD3)2SO) δ 7.81 - 7.59 (m, 5H), 7.13 - 6.93 (m, 4H), 4.80 (d, J = 4.1 Hz, 1H), 3.81 - 3.57 (m, 5H), 2.87 - 2.64 (m, 4H), 2.41 - 2.21 (m, 2H), 2.04 (t, J = 11.9 Hz, 1H), 1.81 - 1.72 (m, 1H), 1.63 - 1.48 (m, 1H).

[0538] Example 378: Preparation of 1-(4-((6-(3R,4R,5S-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3,5-dihydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0539] Step 1: Preparation of (3aR,5R,6R,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxolane-6-yl trifluoromethanesulfonate

Chemical Structure

[0540] (3aR,5S,6R,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxolane-6-ol (10 g, 3.842 mmol, 1.0 equiv.) and pyridine (11.55 g, 14.6 mmol, 3.8 equiv.) were dissolved in dichloromethane (600 mL), and the reaction system was cooled to -17°C. Trifluoromethanesulfonic anhydride (13 g, 4.61 mmol, 1.2 equiv.) was placed in a 100 mL normal pressure dropping funnel and added dropwise to the reaction system at -20°C. After the addition was complete, the reaction was continued at -20°C for 2 hours. After the reaction was completed, the reaction solution was poured into saturated NaHCO3 in ice (100 mL), extraction was performed, the layers were separated, and the aqueous phase was further washed twice with methylene chloride (50 mL / time). The organic phases were combined, dried over Na2SO4, and suction filtered. The filtrate was concentrated, and toluene (150 mL) was added to the residue and concentrated. This operation was performed twice. n-Hexane (160 mL) was added to the residue. The n-hexane solution was filtered. The filtrate was further concentrated to obtain the title compound (14.72 g, yield 97.7%). 11H NMR (400 MHz, CDCl3) δ 5.84 (d, J = 3.8 Hz, 1H), 4.92-4.89 (m, 1H), 4.78-4.76 (m, 1H), 4.23-4.10 (m, 3H), 3.93-3.89 (m, 1H), 1.59 (s, 3H), 1.45 (s, 3H), 1.39 (s, 3H), 1.35 (s, 3H).

[0541] Step 2: Preparation of (3aR,5S,6S,6aR)-6-azido-5-((R)-2,2-dimethyl-1,3-dioxol-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxole [Chemical Structure Diagram]

[0542] (3aR,5R,6R,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxolan-6-yl trifluoromethanesulfonate (14.7 g, 37.47 mmol, 1.0 equiv.) was dissolved in DMF (100 mL), and sodium azide (4.87 g, 74.94 mmol, 2.0 equiv.) was added to the system. The reaction solution was reacted at 90 °C for 16 hours, and the completion of the reaction was confirmed by TLC (petroleum ether:ethyl acetate = 10:1). Most of the reaction solution was concentrated, and 100 mL of ethyl acetate was added to this solution. The ethyl acetate solution was extracted with water (washed 3 times with 50 mL each time). The organic phase was washed once with saturated brine (100 mL). The organic phase was dried over Na2SO4 and filtered by suction. The organic phase was concentrated to obtain a crude product. The crude product was subjected to silica gel column chromatography (normal phase) (PE:EA = 10:1) to obtain the title compound (8.45 g, yield 79.1%).

[0543] 11H NMR (400 MHz, CDCl3) δ 5.86 (d, J = 3.6 Hz, 1H), 4.62 (d, J = 3.6 Hz, 1H), 4.28 - 4.21 (m, 1H), 4.17 - 4.07 (m, 3H), 4.00 - 3.97 (m, 1H), 1.51 (s, 3H), 1.44 (s, 3H), 1.37 (s, 3H), 1.33 (s, 3H).

[0544] Step 3: Preparation of (3aR,5S,6S,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxolan-6-amine

Chemical Structure

[0545] (3aR,5S,6S,6aR)-6-Azido-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxole (200 mg, 0.701 mol, 1 equiv.) was dissolved in methanol (5 mL), and 50 mg of 10% Pd / C was added. The reaction mixture was stirred at 16 °C for 3 h under hydrogen. The completion of the reaction was monitored by TLC and LC-MS. Pd / C was filtered off, and the filtrate was dried with a rotary evaporator under reduced pressure and purified by column chromatography (DCM:MeOH = 98:2) to give the title compound (160 mg, 88% yield).

[0546] LC-MS (ESI) [M+H] + = 260.2; 11H NMR (400 MHz, ) δ 5.89 (d, J = 3.6 Hz, 1H), 4.40 (d, J = 3.6 Hz, 1H), 4.23 - 4.18 (m, 1H), 4.18 - 4.14 (m, 1H), 4.06 - 4.01 (m, 1H), 4.00 - 3.96 (m, 1H), 3.56 (d, J = 3.4 Hz, 1H), 1.51 (s, 3H), 1.42 (s, 3H), 1.35 (s, 3H), 1.31 (s, 3H).

[0547] Step 4: Preparation of 2 - ((3aR,5S,6S,6aR) - 5 - ((R) - 2,2 - dimethyl - 1,3 - dioxolan - 4 - yl) - 2,2 - dimethyltetrahydrofuro[2,3 - d][1,3]dioxolan - 6 - yl) - 1,2,3,4 - tetrahydroisoquinoline

Chemical Structure

[0548] (3aR,5S,6S,6aR) - 5 - ((R) - 2,2 - dimethyl - 1,3 - dioxolan - 4 - yl) - 2,2 - dimethyltetrahydrofuro[2,3 - d][1,3]dioxolan - 6 - amine (60 mg, 0.232 mmol, 1.0 equiv.) was dissolved in methanol (1 mL), and 2 - (2 - bromoethyl)benzaldehyde (54 mg, 0.255 mmol, 1.1 equiv.) was added. The mixture was stirred at 16 °C under N2 protection for 1 hour. The formation of the intermediate was detected by LC - MS, and sodium cyanoborohydride (41 mg, 0.696 mmol, 3.0 equiv.) was further added at 0 °C, and stirring was continued at 16 °C for 1 hour. The completion of the reaction was monitored by TLC and LC - MS. An appropriate amount of water and dichloromethane were added to the reaction solution, and the organic layer was washed once with water and once with saturated NaCl solution, and dried over anhydrous sodium sulfate. The solvent was dried with a rotary evaporator and purified by column chromatography (PE:EA = 2:1) to obtain the title compound (18 mg, yield 20.7%).

[0549] LC-MS (ESI) [M+H] + = 376.2; 1 H NMR (400 MHz, CDCl3) δ 7.08-6.99 (m, 3H), 6.97-6.89 (m, 1H), 5.78 (d, J = 3.8 Hz, 1H), 4.79 (d, J = 3.7 Hz, 1H), 4.33-4.25 (m, 1H), 4.14-4.07 (m, 1H), 4.06-3.95 (m, 2H), 3.88 (d, J = 14.8 Hz, 1H), 3.70 (d, J = 14.8 Hz, 1H), 3.34 (d, J = 4.7 Hz, 1H), 3.01-2.91 (m, 1H), 2.88-2.79 (m, 1H), 2.78-2.61 (m, 2H), 1.46 (s, 3H), 1.35 (s, 3H), 1.26 (s, 3H), 1.25 (s, 3H).

[0550] Step 5: Preparation of (R)-1-((3aR,5S,6S,6aR)-6-(3,4-dihydroisoquinolin-2(1H)-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl)ethane-1,2-diol

Chem.

[0551] 2-((3aR,5S,6S,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-yl)-1,2,3,4-tetrahydroisoquinoline (180 mg, 0.48 mmol, 1.0 equiv.) was dissolved in methanol (1 mL), and 1% H2SO4 (1 mL) was added. The mixture was stirred at 40 °C for 16 h. The formation of the target product was detected by LC-MS. The reaction solution was adjusted to pH = 8 with saturated NaHCO3 solution, and the solvent was dried with a rotary evaporator and purified by column chromatography (DCM:MeOH = 98:2) to obtain the title compound (82 mg, yield 51%).

[0552] LC-MS (ESI) [M+H] + = 336.2; 1 1H NMR (400 MHz, CD3OD) δ 7.12 - 7.05 (m, 3H), 7.05 - 7.00 (m, 1H), 5.92 (d, J = 3.8 Hz, 1H), 5.01 (d, J = 3.8 Hz, 1H), 4.23 - 4.12 (m, 1H), 4.01 - 3.90 (m, 2H), 3.83 - 3.73 (m, 2H), 3.65 - 3.56 (m, 1H), 3.45 (d, J = 5.0 Hz, 1H), 3.21 - 3.09 (m, 1H), 2.99 - 2.87 (m, 1H), 2.86 - 2.71 (m, 2H), 1.48 (s, 3H), 1.32 (s, 3H).

[0553] Step 6: Preparation of (3aR,5S,6S,6aR)-6-(3,4-dihydroisoquinolin-2(1H)-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxole-5-carbaldehyde

Chemical Structure

[0554] (R)-1-((3aR,5S,6S,6aR)-6-(3,4-dihydroisoquinolin-2(1H)-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl)ethane-1,2-diol (60 mg, 0.15 mmol, 1.0 equiv.) was dissolved in acetone / water (1.2 mL, 5:1), and NaIO4 (48 mg, 0.224 mmol, 1.5 equiv) was added at 0 °C. Stirring was continued at 16 °C for 1 h, and cyclobutanecarbonyl chloride (21.29 mg, 0.18 mmol, 1.1 equiv) was added dropwise under ice bath conditions and stirred for 1 h. TLC (PE:EA = 2:1, H2SO4 / EtOH as the color developer, R f=0.5) for the completion of the reaction, the solvent was dried on a rotary evaporator, the residue was dissolved in dichloromethane and suction filtered to obtain a filtrate, and the solvent was dried on a rotary evaporator to obtain a crude product, which was directly used in the next step.

[0555] Seventh step: Preparation of ((3aR,5S,6S,6aR)-6-(3,4-dihydroisoquinolin-2(1H)-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl)methanol [ka]

[0556] The crude (3aR,5S,6S,6aR)-6-(3,4-dihydroisoquinolin-2(1H)-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxole-5-carbaldehyde (50 mg, 0.165 mmol, 1.0 equiv.) was dissolved in methanol / water (1 mL, 4:1), NaBH4 (10 mg, 0.248 mmol, 1.5 equiv) was added at 0°C, and stirring was continued at 16°C for 0.5 h. The reaction was monitored for completion by TLC (DCM:MeOH =20:1, H2SO4 / EtOH as developer), and the reaction was added with saturated ammonium chloride solution (1 mL) and extracted with DCM. The organic layer was washed once each with water and saturated NaCl solution, and dried over anhydrous sodium sulfate. The solvent was dried by rotary evaporation and purified by column chromatography (DCM:MeOH = 98:2) to give the target compound (32 mg, yield 63.5%).

[0557] LC-MS (ESI) [M+H] + = 306.1; 11H NMR (400 MHz, CDCl3) δ 7.13 (dt, J = 8.5, 4.7 Hz, 3H), 7.06 - 7.00 (m, 1H), 5.97 (d, J = 4.0 Hz, 1H), 4.91 (d, J = 3.3 Hz, 1H), 4.49 (q, J = 5.7 Hz, 1H), 4.00 (dd, J = 12.1, 6.0 Hz, 2H), 3.88 (dd, J = 12.2, 5.8 Hz, 1H), 3.75 (d, J = 14.3 Hz, 1H), 3.43 (d, J = 4.1 Hz, 1H), 3.16 (s, 2H), 2.93 (d, J = 7.2 Hz, 1H), 2.88 - 2.68 (m, 2H), 1.54 (s, 3H), 1.36 (s, 3H).

[0558] Step 8: Preparation of ((3aR,5S,6S,6aR)-6-(3,4-dihydroisoquinolin-2(1H)-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl)methyl 4-methylbenzenesulfonate

Chemical Structure

[0559] ((3aR,5S,6S,6aR)-6-(3,4-Dihydroisoquinolin-2(1H)-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl)methanol (250 mg, 0.819 mmol, 1.0 equiv.) was dissolved in dichloromethane (4 mL), and p-methylbenzenesulfonyl chloride (312.2 mg, 1.64 mmol, 2.0 equiv) and pyridine (0.5 mL) were added, and the mixture was stirred at 16 °C for 16 h. The solvent was dried by rotary evaporator and purified by column chromatography (PE:EA = 93:7) to obtain the title compound (125 mg, yield 33.2%).

[0560] LC-MS (ESI) [M+H] += 460.4; 1 1H NMR (400 MHz, CDCl3) δ 7.77 - 7.71 (m, 2H), 7.18 - 7.12 (m, 4H), 7.08 - 7.04 (m, 1H), 7.01 - 6.94 (m, 1H), 5.85 (d, J = 3.7 Hz, 1H), 4.87 - 4.78 (m, 1H), 4.54 - 4.47 (m, 1H), 4.37 - 4.28 (m, 1H), 4.26 - 4.18 (m, 1H), 3.93 - 3.81 (m, 1H), 3.74 - 3.54 (m, 1H), 3.45 - 3.28 (m, 1H), 3.00 - 2.85 (m, 1H), 2.75 - 2.49 (m, 3H), 2.35 (s, 3H), 1.50 (s, 3H), 1.32 (s, 3H).

[0561] Step 9: Preparation of 2 - ((3aR,5R,6S,6aR) - 5 - (azidomethyl) - 2,2 - dimethyltetrahydrofuro[2,3 - d][1,3]dioxol - 6 - yl) - 1,2,3,4 - tetrahydroisoquinoline

Chem.

[0562] ((3aR,5S,6S,6aR) - 6 - (3,4 - dihydroisoquinolin - 2(1H) - yl) - 2,2 - dimethyltetrahydrofuro[2,3 - d][1,3]dioxol - 5 - yl)methyl 4 - methylbenzenesulfonate (126 mg, 0.274 mmol, 1.0 equiv.) was dissolved in N,N - dimethylformamide (1.0 mL), sodium azide (54 mg, 0.822 mmol, 3.0 equiv.) was added, and the mixture was stirred at 90 °C for 16 h under nitrogen. The completion of the reaction was monitored by TLC and LC - MS. Water and ethyl acetate were added to the reaction system, and the organic layer was washed once with water and saturated NaCl solution, and dried over anhydrous sodium sulfate. The solvent was dried by rotary evaporator and purified by column chromatography (PE:EA = 2:1) to obtain the title compound (62 mg, yield 68.4%). LC-MS (ESI) [M + H] + = 331.1。

[0563] Step 10: Preparation of (2S,3R,4R,5R)-5-(azidomethyl)-4-(3,4-dihydroisoquinolin-2(1H)-yl)tetrahydrofuran-2,3-diol

Chem.

[0564] Trifluoroacetic acid / water (0.5 mL, 3:2) was added to 2-((3aR,5R,6S,6aR)-5-(azidomethyl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-yl)-1,2,3,4-tetrahydroisoquinoline (100 mg, 0.302 mmol, 1.0 equiv.), and the reaction mixture was stirred at 16 °C for 24 h. The completion of the reaction was monitored by TLC and LC-MS. A formazan-based drying solvent was added to the system and used directly in the next reaction without further purification.

[0565] Step 11: Preparation of (3S,4R,5R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-3,5-diol

Chem.

[0566] (2S,3R,4R,5R)-5-(azidomethyl)-4-(3,4-dihydroisoquinolin-2(1H)-yl)tetrahydrofuran-2,3-diol (85 mg, 0.292 mmol, 1.0 equiv.) was added to methanol (10 mL) and 10% Pd / C (30 mg), and the reaction mixture was stirred at 16 °C under hydrogen for 8 h. The completion of the reaction was monitored by TLC and LC-MS. After filtration, the solvent was dried using a rotary evaporator, and the crude product was separated and purified by Prep-HPLC (C18, 10 mmol / L aqueous solution, MeCN) to obtain the target compound (67 mg).

[0567] LC-MS (ESI) [M+H] + = 249.1; 1 1H NMR (400 MHz, CDCl3) δ 7.18 - 7.09 (m, 3H), 7.06 - 6.98 (m, 1H), 4.10 (s, 2H), 3.79 - 3.65 (m, 2H), 3.28 (dd, J = 11.5, 4.6 Hz, 2H), 3.21 - 3.11 (m, 2H), 2.96 - 2.87 (m, 2H), 2.62 - 2.47 (m, 4H), 2.43 (t, J = 9.9 Hz, 1H).

[0568] Step 12: Preparation of 1-(4-((6-(3R,4R,5S)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3,5-dihydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chem.

[0569] (3S,4R,5R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)piperidine-3,5-diol (50 mg, 0.2 mmol, 1.0 equiv.), 6-((1-acetylpiperidin-4-yl)amino)pyrimidine-4-carboxylic acid (58 mg, 0.22 mmol, 1.1 equiv.), and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (114 mg, 0.3 mmol, 1.5 equiv.) were added to N,N-dimethylformamide (0.6 mL), and further N,N-diisopropylethylamine (78 mg, 0.6 mmol, 3.0 equiv.) was added. The mixture was stirred at 16 °C for 0.5 h, and the completion of the reaction of the reactants was detected by TLC. Water (15 mL) was added to the reaction system, and the mixture was extracted three times with ethyl acetate (3 × 15 mL). The combined organic phases were washed once with a saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered by suction, and the solvent was concentrated. The target compound was obtained by separation and purification by Prep-HPLC (C18, 10 mmol / L aqueous solution, MeCN) (18.29 mg, yield: 18.6%).

[0570] LC-MS (ESI) [M+H] + = 495.3; 1 1H NMR (400 MHz, CDCl3) δ 8.53 (s, 1H), 7.19 - 7.09 (m, 3H), 7.04 - 6.98 (m, 1H), 6.69 (s, 1H), 5.48 (s, 1H), 4.63 - 4.49 (m, 2H), 4.23 - 3.95 (m, 5H), 3.92 - 3.77 (m, 2H), 3.30 - 3.08 (m, 4H), 3.05 - 2.88 (m, 3H), 2.86 - 2.67 (m, 2H), 2.17 - 2.12 (m, 1H), 2.11 (s, 3H), 2.06 - 1.98 (m, 1H), 1.45 (m, 2H).

[0571] Example 379: Preparation of trans-1-(4-((6-(4-amino-3-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0572] Step 1: Preparation of benzyl 4-amino-3-hydroxypiperidine-1-carboxylate and benzyl 3-amino-4-hydroxypiperidine-1-carboxylate

Chemical formula

[0573] Benzyl 7-oxa-3-azabicyclo[4.1.0]heptane-3-carboxylate (2.5 g, 10.72 mmol, 1 equiv.) was dissolved in ethanol (5 mL), and concentrated aqueous ammonia (15 mL, 25 - 28%, 210 mmol, 19.6 equiv.) was added. The mixture was stirred at 70 °C overnight (16 hours). The completion of the reaction was monitored by TLC (PE:EA = 2:1). The reaction solution was centrifuged to obtain a crude product (2.8 g), which was used directly in the next step. LC-MS (ESI) [M + H] + = 251.2.

[0574] Step 2: Preparation of benzyl 4-((tert-butoxycarbonyl)amino)-3-hydroxypiperidine-1-carboxylate and benzyl 3-((tert-butoxycarbonyl)amino)-4-hydroxypiperidine-1-carboxylate

Chemical formula

[0575] The crude product from the previous step (2.8 g, 10.72 mmol, 1 equiv.) was dissolved in dichloromethane (25 mL), and triethylamine (1.63 g, 16.11 mmol, 1.5 equiv.) and di-tert-butyl dicarbonate (2.8 g, 12.83 mmol, 1.2 equiv.) were added, followed by stirring at room temperature for 1 hour. The reaction was monitored by LC-MS, and the reaction solution was used in the next step without purification. LC-MS (ESI) [ (M-100) +1] + = 251.20, [ (M-56) +1] + = 295.20.

[0576] Step 3: Preparation of benzyl 4-((tert-butoxycarbonyl)amino)-3-(p-toluenesulfonyloxy)piperidine-1-carboxylate and benzyl 3-((tert-butoxycarbonyl)amino)-4-(p-toluenesulfonyloxy)piperidine-1-carboxylate

Chemical Structure

[0577] To the reaction solution from the previous step (10.72 mmol of raw material), triethylamine (2.17 g, 21.44 mmol, 2 equiv.), 4-toluenesulfonyl chloride (4.09 g, 21.45 mmol, 2 equiv.) and 4-dimethylaminopyridine (131 mg, 1.072 mmol, 0.1 equiv.) were sequentially added, and the mixture was stirred at room temperature for 16 hours. The TLC monitoring showed the completion of the reaction. The solvent was centrifuged off, and the crude product was purified by flash chromatography (silica gel, PE solution with 0-27% EA) to obtain the product (3.317 g, total yield of three steps: 61.4%).

[0578] LC-MS (ESI) [M+H] + = 348.0; 11H NMR (400 MHz, CDCl3) δ 7.88 - 7.66 (m, 2H), 7.43 - 7.17 (m, 7H), 5.16 - 5.04 (m, 2H), 4.73 - 4.44 (m, 1H), 4.32 - 4.08 (m, 1H), 4.04 - 3.76 (m, 1H), 3.74 - 3.54 (m, 1H), 3.43 - 2.75 (m, 2H), 2.49 - 2.34 (m, 3H), 2.16 - 2.07 (m, 0.7H), 1.96 - 1.85 (m, 0.3H), 1.51 - 1.35 (m, 10H).

[0579] Step 4: Preparation of 3-benzyl 7-tert-butyl 3,7-diazabicyclo[4.1.0]heptane-3,7-dicarboxylate

Chem.

[0580] The product of the third step (3.317 g, 6.574 mmol, 1 equiv.) was dissolved in DMF (28 mL), an aqueous sodium hydroxide solution (526 mg / 7 mL, 13.15 mmol, 2 equiv.) was added, and the mixture was heated and stirred at 50 °C for 1 hour. The completion of the reaction was monitored by LC-MS. Ethyl acetate was added to the reaction solution for dilution, washed with water, separated, the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, dried on a rotary evaporator, and the crude product was separated and purified by flash chromatography (silica gel, PE with 0 - 20% EA) to obtain the title compound (2.0 g, yield 91.5%) as a colorless oil. LC-MS (ESI) [(M - 100) + 1] + = 233.20, [(M - 56) + 1] + = 277.20.

[0581] Step 5: Preparation of benzyl trans-4-((tert-butoxycarbonyl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate

Chem.

[0582] 3-Benzyl 7-tert-butyl 3,7-diazabicyclo[4.1.0]heptane-3,7-dicarboxylate (200 mg, 0.602 mmol, 1 equiv.) and 1,2,3,4-tetrahydroisoquinoline (120 mg, 0.901 mmol, 1.5 equiv.) were dissolved in toluene (3 mL), and tributylphosphine (13 mg, 0.0643 mmol, 0.11 equiv.) was added. The reaction mixture was refluxed overnight (16 h) under nitrogen. The residue of the starting material was monitored by LC-MS. Tetrahydroisoquinoline (96 mg, 0.722 mmol, 1.2 equiv.) and tributylphosphine (24 mg, 0.120 mmol, 0.2 equiv.) were added, and the mixture was reacted at 110 °C for 1 day (24 h). The solvent was dried by rotary evaporator, and the crude product was separated and purified by flash chromatography (silica gel, PE solution with 0 - 15% EA), and crude benzyl trans-4-((tert-butoxycarbonyl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate (166 mg) was obtained. It was purified by reverse-phase column (C18, 10 mmol / L aqueous NH4HCO3 solution, 30% - 90% MeCN) to obtain the title compound (97 mg, yield 34.7%).

[0583] LC-MS (ESI) [M+H] + = 466.2; 1 H NMR (400 MHz, DMSO-d6) δ 7.44 - 7.26 (m, 5H), 7.11 - 6.93 (m, 4H), 6.61 (d, J = 8.8 Hz, 1H), 5.09 (s, 2H), 4.18 - 4.04 (m, 1H), 3.97 - 3.82 (m, 2H), 3.80 - 3.63 (m, 2H), 3.11 - 3.00 (m, 1H), 2.97 - 2.57 (m, 5H), 2.54 - 2.46 (m, 1H), 1.86 - 1.76 (m, 1H), 1.45 - 1.25 (m, 10H).

[0584] Step 6: Preparation of tert-butyl trans-(3-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-4-yl)carbamate

Chem.

[0585] Benzyl trans-4-((tert-butoxycarbonyl)amino)-3-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate (87 mg, 0.187 mmol, 1 equiv.) was dissolved in methanol (chromatography grade, 5 mL), and stirred under Pd / C (10%, 90 mg), H2 conditions for 1.5 h. The reaction was monitored by TLC, and the reaction solution was filtered and centrifuged to obtain the crude product (60 mg, yield 96.9%). The crude product was used directly in the next step. LC-MS (ESI) [M + H] + = 332.25.

[0586] Step 7: Preparation of tert-butyl trans-(1-(6-((1-acetylpiperidin-4-yl)amino)pyrimidine-4-carbonyl)-3-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-4-yl)carbamate

Chem.

[0587] 6 - ((1 - acetylpiperidin - 4 - yl)amino)pyrimidine - 4 - carboxylic acid (48 mg, 0.182 mmol, 1 equiv.), EDCI (53 mg, 0.276 mmol, 1.52 equiv.) and HOAt (37 mg, 0.271839 mmol, 1.5 equiv.) were dissolved in DMF (2 mL) and stirred for 5 minutes. Then, a DMF solution (1 mL) of tert - butyl trans - (3 - (3,4 - dihydroisoquinolin - 2(1H) - yl)piperidin - 4 - yl)carbamate (45 mg, 0.174 mmol, 1.7 equiv.) was added. The reaction mixture was stirred at room temperature for 1.5 hours. The solvent was dried by rotary evaporator, and the crude product was separated and purified by flash chromatography (silica gel, DCM solution with 0 - 5% MeOH) to obtain the title compound (115 mg, purity 80%, yield 91.6%). LC - MS (ESI) [ M + H] + = 578.2。

[0588] Step 8: Preparation of trans - 1 - (4 - ((6 - (4 - amino - 3 - (3,4 - dihydroisoquinolin - 2(1H) - yl)piperidine - 1 - carbonyl)pyrimidine - 4 - yl)amino)piperidine - 1 - yl)ethan - 1 - one

Chemical formula

[0589] tert-Butyl trans-(1-(6-((1-acetylpiperidin-4-yl)amino)pyrimidine-4-carbonyl)-3-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-4-yl)carbamate (115 mg, 80%, 0.159 mmol, 1 equiv.) was dissolved in dichloromethane (4 mL), trifluoroacetic acid (2 mL) was added, and the mixture was stirred at room temperature for 1 hour. The completion of the reaction was monitored by LC-MS, dichloromethane was dried with trifluoroacetic acid on a rotary evaporator, the crude product was dissolved in methanol, a strongly basic anion exchange resin was added to adjust the pH to 8 - 9, filtered, dried on a rotary evaporator, and purified by a reverse-phase column (C18, 10 mmol / L aqueous NH4OH / acetonitrile) to obtain the title compound (19.46 mg, yield: 25.1%).

[0590] LC-MS (ESI) [M+H] + = 478.4; 1 H NMR (400 MHz, CDCl3) δ 8.58 - 8.51 (m, 1H), 7.18 - 6.95 (m, 4H), 6.68 - 6.56 (m, 1H), 5.34 - 5.13 (m, 1H), 4.93 - 4.66 (m, 1H), 4.61 - 4.50 (m, 1H), 4.27 - 4.20 (m, 0.4H), 4.13 - 4.08 (m, 0.6H), 4.05 - 3.97 (m, 0.6H), 3.91 - 3.70 (m, 2.4H), 3.28 - 3.03 (m, 3H), 3.03 - 2.88 (m, 2H), 2.87 - 2.61 (m, 4.4H), 2.55 - 2.46 (m, 0.6H), 2.19 - 1.95 (m, 6H), 1.58 - 1.35 (m, 3H).

[0591] Example 380: Preparation of 1-(4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-carbonyl)-2-(isopropylsulfinyl)pyrimidine-4-yl)amino)piperidin-1-yl)ethan-1-one [Chemistry]

[0592] 1-(4-((6-((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-(isopropylthio)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one (40 mg, 0.072368 mmol, 1 equiv.) and m-CPBA (13.7374 mg, 0.079605 mmol, 1.1 equiv.) were dissolved in dichloromethane (2 mL), and the reaction mixture was stirred at room temperature for 2 h. The reaction solution was dried to dryness by a rotary evaporator to obtain a crude product, which was separated and purified by reverse-phase HPLC (C18, aqueous NH4HCO3 / acetonitrile) to obtain the title compound (15 mg).

[0593] LC-MS (ESI) [M+H] + = 569.0; 1 H NMR (400 MHz, DMSO) δ 8.48 - 7.45 (m, 1H), 7.17 - 6.99 (m, 4H), 6.81 - 6.56 (m, 1H), 4.85 - 4.69 (m, 1H), 4.46 - 4.09 (m, 3H), 3.86 - 3.76 (m, 3H), 3.69 - 3.59 (m, 2H), 3.26 - 3.13 (m, 2H), 3.08 - 2.73 (m, 6H), 2.70 - 2.59 (m, 2H), 2.07 - 1.66 (m, 6H), 1.55 - 1.25 (m, 6H), 1.11 - 0.96 (m, 2H).

[0594] Examples 381 - 384 Using the method of Example 378, the compounds of Examples 381 - 384 were synthesized, and the structures and specific identification data (LC-MS and 1 H NMR) of the compounds were as follows.

Table 7

[0595] Example 385 and Example 386: Preparation of (E)-1-(4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)-2-(methoxyimino)propan-1-one (Compound of Example 385) and (Z)-1-(4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)-2-(methoxyimino)propan-1-one (Compound of Example 386)

[0596] Step 1: Preparation of tert-butyl (1-(2-oxopropanoyl)piperidin-4-yl)carbamate [Chemical formula]

[0597] tert-Butyl piperidin-4-ylcarbamate (1 g, 5 mmol, 1 equiv.), EDCI (1.15 g, 6 mmol, 1.2 equiv.), HOAt (0.815 g, 6 mmol, 1.2 equiv.) were dissolved in DMF (10 mL), 2-oxopropanoic acid (0.44 g, 5 mmol, 1.0 equiv.) was added, and the mixture was stirred at room temperature (20 °C) for 1 hour. After monitoring the completion of the reaction by LC-MS, the mixture was concentrated, 20 mL of water was added, and the mixture was extracted three times with 20 mL of ethyl acetate each time. The organic phases were combined and dried on a rotary evaporator to obtain a crude product. The crude product was purified by column chromatography (PE:EA = 1:1) to obtain the title compound (1.1 g, yield: 81.5%). LC-MS (ESI) [M + H] + = 215.0.

[0598] Step 2: Preparation of tert-butyl (1-(2-(methoxyimino)propionyl)piperidin-4-yl)carbamate [Chemical formula]

[0599] tert-Butyl (1-(2-oxopropanoyl)piperidin-4-yl)carbamate (600 mg, 2.2 mmol, 1 equiv.), methoxyamine hydrochloride (407.8 mg, 4.8 mmol, 2.2 equiv.) were dissolved in ethanol (6 mL) and water (1.5 mL), and potassium acetate (958 mg, 9.6 mmol, 4.4 equiv.) was added. The reaction mixture was stirred at 80 °C for 16 h at room temperature. After completion of the reaction by LC-MS, it was concentrated, 20 mL of water was added, and extraction was performed 3 times with 20 mL of ethyl acetate each time. The organic phases were combined, dried with a rotary evaporator to obtain a crude product, and purified by column chromatography to obtain tert-butyl (E)-(1-(2-(methoxyimino)propionyl)piperidin-4-yl)carbamate (290 mg) and tert-butyl (Z)-(1-(2-(methoxyimino)propionyl)piperidin-4-yl)carbamate (337 mg). LC-MS (ESI) [M + H] + = 244.0.

[0600] Step 3: Preparation of 1-(4-aminopiperidin-1-yl)-2-(methoxyimino)propan-1-one [Chemical formula]

[0601] tert-Butyl (E)-(1-(2-(methoxyimino)propionyl)piperidin-4-yl)carbamate and tert-butyl (Z)-(1-(2-(methoxyimino)propionyl)piperidin-4-yl)carbamate (600 mg, 2 mmol, 1 equiv.) were dissolved in 1,4-dioxane (5 mL), and 1,4-dioxane solution of hydrochloric acid (2.5 mL, 10 mmol, 4 M) was added. The mixture was stirred at 50 °C for 1 hour at room temperature. After monitoring the completion of the reaction by LC-MS, it was concentrated and dried by a rotary evaporator to obtain 584 mg of a crude product, which was directly used in the next step. LC-MS (ESI) [ M + H] + = 200.2.

[0602] Step 4: Preparation of methyl 6-((1-(2-(methoxyimino)propionyl)piperidin-4-yl)amino)pyrimidine-4-carboxylate

Chemical Structure

[0603] 1-(4-Aminopiperidin-1-yl)-2-(methoxyimino)propan-1-one (550 mg, 1.96 mmol, 1 equiv.) and methyl 6-chloropyrimidine-4-carboxylate (338 mg, 1.96 mmol, 1.0 equiv.) were dissolved in acetonitrile (6 mL), and DIPEA (1.01 g, 7.84 mmol, 4.0 equiv.) was added. The reaction mixture was stirred at 90 °C for 2 hours at room temperature. After monitoring the completion of the reaction by LC-MS, it was concentrated, 20 mL of water was added, and the mixture was extracted three times with 20 mL of ethyl acetate each time. The combined organic phases were dried by a rotary evaporator to obtain a crude product, which was purified by column chromatography to obtain the target compound (438 mg, yield 66%). LC-MS (ESI) [ M + H] + = 336.2.

[0604] Step 5: Preparation of 6 - ((1 - (2 - (methoxyimino)propionyl)piperidin - 4 - yl)amino)pyrimidine - 4 - carboxylic acid

Chem.

[0605] Methyl 6 - ((1 - (2 - (methoxyimino)propionyl)piperidin - 4 - yl)amino)pyrimidine - 4 - carboxylate (470 mg, 1.4 mmol, 1 equiv.) was dissolved in THF (4 mL) and H2O (4 mL), lithium hydroxide (2.8 mL, 2.8 mmol, 2.0 equiv., 2 M) was added, the reaction mixture was stirred at room temperature (25 °C) for 1 hour, after monitoring the completion of the reaction by LC - MS, 1 M aqueous hydrochloric acid was added to adjust the pH to 5 - 6, concentrated, and dried on a rotary evaporator to obtain the crude product (450 mg), which was used directly in the next step without further purification. LC - MS (ESI) [M + H] + = 322.2.

[0606] Step 6: Preparation of (E) - 1 - (4 - ((6 - (3R,4R - 4 - (3,4 - dihydroisoquinolin - 2(1H) - yl) - 3 - hydroxypiperidine - 1 - carbonyl)pyrimidin - 4 - yl)amino)piperidin - 1 - yl) - 2 - (methoxyimino)propan - 1 - one (compound of Example 385) and (Z) - 1 - (4 - ((6 - (3R,4R - 4 - (3,4 - dihydroisoquinolin - 2(1H) - yl) - 3 - hydroxypiperidine - 1 - carbonyl)pyrimidin - 4 - yl)amino)piperidin - 1 - yl) - 2 - (methoxyimino)propan - 1 - one (compound of Example 386)

Chem.

[0607] 6 - ((1 - (2 - (Methoxyimino)propionyl)piperidin - 4 - yl)amino)pyrimidine - 4 - carboxylic acid (420 mg, 1.3 mmol, 1 equiv.), EDCI (325 mg, 1.7 mmol, 1.3 equiv.), HOAt (231 mg, 1.7 mmol, 1.3 equiv.) were dissolved in DMF (5 mL), (3R,4R) - 4 - (3,4 - dihydroisoquinolin - 2(1H) - yl)piperidin - 3 - ol (334 mg, 1.4 mmol, 1.1 equiv.) was added, and the reaction mixture was stirred at room temperature (20 °C) for 1 hour. After monitoring the completion of the reaction by LC - MS, it was concentrated, and the crude product was separated and purified by reverse - phase HPLC (10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain two compounds:

[0608] The title compound (compound of Example 385) (184 mg). LC - MS (ESI) [M + H] + = 536.0; 1 H NMR (400 MHz, (CD3)2SO) δ 8.45 - 8.43 (m, 1H), 7.69 - 7.63 (m, 1H), 7.09 - 7.02 (m, 4H), 6.53 (s, 1H), 4.81 - 4.72 (m, 1H), 4.49 - 4.31 (m, 1H), 4.18 - 4.13 (m, 2H), 3.85 - 3.58 (m, 7H), 3.47 - 3.40 (m, 1H), 3.20 - 3.14 (m, 1H), 3.02 - 2.55 (m, 8H), 1.95 - 1.93 (m, 5H), 1.85 - 1.71 (m, 1H), 1.52 - 1.30 (m, 3H).

[0609] The title compound (compound of Example 386) (301.72 mg). LC - MS (ESI) [M + H] + = 536.0; 11H NMR (400 MHz, (CD3)2SO) δ 8.45 - 8.43 (m, 1H), 7.68 - 7.64 (m, 1H), 7.09 - 7.02 (m, 4H), 6.52 (s, 1H), 4.81 - 4.72 (m, 1H), 4.49 - 4.31 (m, 1H), 4.25 - 4.17 (m, 2H), 3.88 - 3.59 (m, 8H), 3.25 - 3.19 (m, 1H), 3.02 - 2.76 (m, 6.5H), 2.67 - 2.55 (m, 1.5H), 1.93 - 1.93 (m, 5H), 1.84 - 1.71 (m, 1H), 1.51 - 1.32 (m, 3H).

[0610] Example 387 and Example 388: Preparation of (E)-1-(4-((6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-carbonyl)pyrimidin-4-yl)amino)-3-azabicyclo[3.1.1]hept-3-yl)-2-(methoxyimino)propan-1-one (Compound of Example 387) and (Z)-1-(4-((6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-carbonyl)pyrimidin-4-yl)amino)-3-azabicyclo[3.1.1]hept-3-yl)-2-(methoxyimino)propan-1-one (Compound of Example 388)

[0611] Step 1: Preparation of tert-butyl (3-(2-oxopropanoyl)-3-azabicyclo[3.1.1]heptan-6-yl)carbamate

Chemical Structure

[0612] tert-Butyl (3-azabicyclo[3.1.1]heptan-6-yl)carbamate (460 mg, 2.17 mmol, 1 equiv.), EDCI (498 mg, 2.6 mmol, 1.2 equiv.) and HOAt (354 mg, 2.6 mmol, 1.2 equiv.) were dissolved in DMF (5 mL), 2-oxopropanoic acid (209.9 mg, 2.38 mmol, 1.1 equiv.) was added, and the reaction mixture was stirred at room temperature (20 °C) for 1 h. After monitoring the completion of the reaction by LC-MS, it was concentrated, 20 mL of water was added, and the mixture was extracted three times with 20 mL of ethyl acetate each time. The combined organic phases were dried on a rotary evaporator to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 1:1) to obtain the title compound (412 mg, yield 67%). LC-MS (ESI) [ M + H] + = 227.0。

[0613] Step 2: Preparation of tert-butyl 3-(2-(methoxyimino)propionyl)-3-azabicyclo[3.1.1]hept-6-yl)carbamate

Chemical formula

[0614] tert-Butyl (3-(2-oxopropionyl)-3-azabicyclo[3.1.1]heptan-6-yl)carbamate (400 mg, 1.4 mmol, 1 equiv.), methoxyamine hydrochloride (260 mg, 3.1 mmol, 2.2 equiv.) were dissolved in ethanol (4 mL) and water (1 mL), and potassium acetate (611.8 mg, 6.2 mmol, 4.4 equiv.) was added. The reaction mixture was stirred at 80 °C for 16 h. After monitoring the completion of the reaction by LC-MS, it was concentrated, 20 mL of water was added, and the mixture was extracted three times with 20 mL of ethyl acetate each time. The combined organic phases were dried on a rotary evaporator to obtain the crude product, which was purified by column chromatography to obtain the target compound (440 mg). LC-MS (ESI) [ M + H]+ = 256.1。

[0615] Step 3: Preparation of 1-(6-amino-3-azabicyclo[3.1.1]hept-3-yl)-2-(methoxyimino)propan-1-one

Chem.

[0616] tert-Butyl (3-(2-(methoxyimino)propionyl)-3-azabicyclo[3.1.1]heptan-6-yl)carbamate (400 mg, 1.28 mmol, 1 equiv.) was dissolved in 1,4-dioxane (5 mL), and 1,4-dioxane solution of hydrochloric acid (2.5 mL, 10 mmol, 4 M) was added. The mixture was stirred at 50 °C for 1 hour at room temperature. After monitoring the completion of the reaction by LC-MS, it was concentrated and dried by a rotary evaporator to obtain 360 mg of a crude product, which was used in the next step as it was. LC-MS (ESI) [ M + H] + = 212.0。

[0617] Step 4: Preparation of methyl 6-((3-(2-(methoxyimino)propionyl)-3-azabicyclo[3.1.1]heptan-6-yl)amino)pyrimidine-4-carboxylate

Chem.

[0618] 1-(6-Amino-3-azabicyclo[3.1.1]heptan-3-yl)-2-(methoxyimino)propan-1-one (320 mg, 1.5 mmol, 1 equiv.) and methyl 6-chloropyrimidine-4-carboxylate (287 mg, 1.6 mmol, 1.1 equiv.) were dissolved in acetonitrile (5 mL), and DIPEA (783 mg, 6.0 mmol, 4.0 equiv.) was added. The reaction mixture was stirred at 90 °C for 2 h, monitored by LC-MS for completion of the reaction, concentrated, 20 mL of water was added, and the mixture was extracted three times with 20 mL portions of ethyl acetate. The combined organic phases were dried over a rotary evaporator to obtain the crude product, which was purified by column chromatography to give the target compound (266 mg, 51% yield). LC-MS (ESI) [ M + H] + = 348.2。

[0619] Step 5: Preparation of 6-((3-(2-(methoxyimino)propionyl)-3-azabicyclo[3.1.1]heptan-6-yl)amino)pyrimidine-4-carboxylic acid

Chemical Structure

[0620] Methyl 6-((3-(2-(methoxyimino)propionyl)-3-azabicyclo[3.1.1]heptan-6-yl)amino)pyrimidine-4-carboxylate (240 mg, 0.69 mmol, 1 equiv.) was dissolved in THF (4 mL) and H2O (4 mL), and lithium hydroxide (0.34 mL, 1.38 mmol, 2.0 equiv., 2 M) was added. The reaction mixture was stirred at 25 °C for 1 h, monitored by LC-MS for completion of the reaction, then 1 M aqueous hydrochloric acid solution adjusted to pH = 5 - 6 was added, concentrated, and dried over a rotary evaporator to obtain 250 mg of the crude product, which was used directly in the next step. LC-MS (ESI) [ M + H] + = 334.20。

[0621] Step 6: Preparation of (E)-1-(4-((6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-carbonyl)pyrimidin-4-yl)amino)-3-azabicyclo[3.1.1]heptan-3-yl)-2-(methoxyimino)propan-1-one (compound of Example 387) and (Z)-1-(4-((6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-carbonyl)pyrimidin-4-yl)amino)-3-azabicyclo[3.1.1]heptan-3-yl)-2-(methoxyimino)propan-1-one (compound of Example 388) [Chemical formula]

[0622] 6-((3-(2-(methoxyimino)propionyl)-3-azabicyclo[3.1.1]heptan-6-yl)amino)pyrimidine-4-carboxylic acid (220 mg, 0.66 mmol, 1 equiv.), EDCI (164.5 mg, 0.86 mmol, 1.3 equiv.), and HOAt (117 mg, 0.86 mmol, 1.3 equiv.) were dissolved in DMF (3 mL), and (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (169 mg, 0.72 mmol, 1.1 equiv.) was added. The reaction mixture was stirred at room temperature (20 °C) for 1 hour. After monitoring the completion of the reaction by LC-MS, the mixture was concentrated, and the crude product was separated and purified by reverse-phase HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain two compounds:

[0623] The title compound (compound of Example 387) (63.15 mg). LC-MS (ESI) [M+H] + = 548.0; 11H NMR (400 MHz, (CD3)2SO) δ 8.47 - 8.45 (m, 1H), 7.63 - 7.59 (m, 1H), 7.09 - 7.01 (m, 4H), 6.71 (s, 1H), 4.82 - 4.70 (m, 1H), 4.48 - 4.07 (m, 2H), 3.85 - 3.70 (m, 5H), 3.62 - 3.40 (m, 6H), 3.03 - 2.85 (m, 2H), 2.81 - 2.51 (m, 7H), 1.93 - 1.93 (m, 4H), 1.85 - 1.70 (m, 1H), 1.53 - 1.32 (m, 2H).

[0624] Title compound (compound of Example 388) (206.44 mg). LC - MS (ESI) [M + H] + = 548.0; 1 1H NMR (400 MHz, (CD3)2SO) δ 8.47 - 8.45 (m, 1H), 7.67 (s, 1H), 7.09 - 7.02 (m, 4H), 6.75 (s, 1H), 4.82 - 4.71 (m, 1H), 4.49 - 4.26 (m, 2H), 3.85 - 3.79 (m, 6H), 3.75 - 3.57 (m, 4H), 3.53 - 3.50 (m, 1H), 3.03 - 2.86 (m, 2H), 2.82 - 2.59 (m, 7H), 1.94 - 1.91 (m, 4H), 1.85 - 1.71 (m, 1H), 1.51 - 1.45 (m, 1H), 1.40 - 1.37 (m, 1H).

[0625] Example 389: Preparation of 1 - (4 - ((4 - benzyl - 5 - ((3R,4R) - 4 - (3,4 - dihydroisoquinolin - 2(1H) - yl) - 3 - hydroxypiperidine - 1 - carbonyl)thiazol - 2 - yl)amino)piperidin - 1 - yl)ethan - 1 - one

[0626] Step 1: Preparation of N - ((1 - acetylpiperidin - 4 - yl)thiocarbamoyl)benzamide

Chemical Structure

[0627] 1-(4-Aminopiperidin-1-yl)ethan-1-one (100 mg, 0.56 mmol, 1.0 equiv.) and benzoyl isothiocyanate (91 mg, 0.56 mmol, 1.0 equiv.) were added to dichloromethane (2.6 mL), and the mixture was stirred at 16 °C for 48 h under nitrogen, and the completion of the reaction was monitored by TLC. Water (15 mL) was added to the reaction system, and the mixture was extracted three times with ethyl acetate (3 × 15 mL). The combined organic phases were washed once with a saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered by suction, the solvent was concentrated, and the residue was purified by column chromatography (DCM:MeOH = 98:2) to obtain the target product (70 mg, yield 32.6%).

[0628] LC-MS (ESI) [M+H] + = 306.2; 1 H NMR (400 MHz, DMSO-d6) δ 11.35 (s, 1H), 10.91 (d, J = 7.6 Hz, 1H), 7.95 - 7.89 (m, 2H), 7.64 (t, J = 7.4 Hz, 1H), 7.51 (t, J = 7.7 Hz, 2H), 4.49 - 4.35 (m, 1H), 4.23 - 4.12 (m, 1H), 3.82 - 3.70 (m, 1H), 3.27 - 3.16 (m, 1H), 2.91 - 2.79 (m, 1H), 2.10 - 2.03 (m, 1H), 2.02 (s, 3H), 2.01 - 1.95 (m, 1H), 1.65 - 1.51 (m, 1H), 1.49 - 1.35 (m, 1H).

[0629] Step 2: Preparation of 1-(1-acetylpiperidin-4-yl)thiourea

Chem.

[0630] N-((1-acetylpiperidin-4-yl)thiocarbamoyl)benzamide (402 mg, 1.32 mmol, 1.0 equiv.) was added to methanol (6.6 mL), and a solution of sodium hydroxide (63 mg, 1.58 mmol, 1.2 equiv., 1.5 M) was added. The mixture was stirred at 80 °C for 2 h, and the completion of the reaction of the starting material was monitored by TLC. The solvent was concentrated to obtain the crude product (410 mg) of the title compound, which was used directly in the next reaction. LC-MS (ESI) [M + H] + = 202.1.

[0631] Step 3: Preparation of methyl 2-chloro-3-oxo-4-phenylbutyrate

Chemical formula

[0632] After methyl 3-oxo-4-phenylbutyrate (100 mg, 0.521 mmol, 1.0 equiv.) was added to dichloromethane (2.6 mL), a mixed solution of sulfonyl chloride (85 mg, 0.625 mmol, 1.2 equiv.) / dichloromethane (1.0 mL) was added dropwise. After stirring at 16 °C for 2 h, the completion of the reaction was monitored by TLC. Water (15 mL) was added to the reaction system, and the mixture was extracted 3 times with ethyl acetate (3 × 15 mL). The organic phases were combined, washed once with a saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered by suction, the solvent was concentrated, and the product was purified by column chromatography (PE:EA = 90:10) to obtain the target product (100 mg, yield 84.8%). 1 H NMR (400 MHz, DMSO-d6) δ 7.36 - 7.30 (m, 2H), 7.29 - 7.26 (m, 1H), 7.23 - 7.17 (m, 2H), 5.76 (s, 1H), 4.07 (s, 2H), 3.75 (s, 3H).

[0633] Step 4: Preparation of Methyl 2-((1-acetylpiperidin-4-yl)amino)-4-benzylthiazole-5-carboxylate

Chem.

[0634] Methyl 2-chloro-3-oxo-4-phenylbutyrate (200 mg, 1.0 mmol, 1.0 equiv.) and 1-(1-acetylpiperidin-4-yl)thiourea (226 mg, 1.0 mmol, 1.0 equiv.) were placed in ethanol (5.0 mL), stirred at 85 °C for 1 hour under nitrogen, and the completion of the reaction was monitored by TLC. Water (15 mL) was added to the reaction system, and the mixture was extracted three times with ethyl acetate (3 × 15 mL). The combined organic phases were washed once with a saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered by suction, the solvent was concentrated, and the product was purified by column chromatography (DCM:MeOH = 98:2) to obtain the target product (140 mg, yield 37.5%).

[0635] LC-MS (ESI) [M+H] + = 374.2; 1 1H NMR (400 MHz, DMSO-d6) δ 8.56 (d, J = 7.4 Hz, 1H), 7.33 - 7.28 (m, 4H), 7.26 - 7.20 (m, 1H), 4.27 (s, 2H), 4.25 - 4.18 (m, 1H), 3.85 - 3.79 (m, 1H), 3.77 (s, 3H), 3.75 - 3.63 (m, 1H), 3.26 - 3.20 (m, 1H), 2.91 - 2.80 (m, 1H), 2.04 (s, 3H), 2.02 - 1.89 (m, 2H), 1.49 - 1.37 (m, 1H), 1.37 - 1.26 (m, 1H).

[0636] Step 5: Preparation of 2-((1-acetylpiperidin-4-yl)amino)-4-benzylthiazole-5-carboxylic Acid

Chem.

[0637] To a solution of methyl 2-((1-acetylpiperidin-4-yl)amino)-4-benzylthiazole-5-carboxylate (22 mg, 0.059 mmol, 1.0 equiv.) in methanol (0.3 mL) was added NaOH solution (9.5 mg, 0.236 mmol, 4.0 equiv., 1 M), and the mixture was stirred at 16 °C for 16 h. The completion of the raw material reaction was detected by TLC. After adjusting the pH to 3 with dilute hydrochloric acid solution, water (15 mL) was added to the reaction system, and the mixture was extracted three times with ethyl acetate (3 × 15 mL). The combined organic phases were washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered by suction, and the solvent was concentrated to obtain the title compound (20 mg, crude product), which was directly used in the next reaction.

[0638] Step 6: Preparation of 1-(4-((4-benzyl-5-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)thiazol-2-yl)amino)piperidin-1-yl)ethan-1-one

Chemical formula

[0639] 2 - ((1 - Acetylpiperidin - 4 - yl)amino) - 4 - benzylthiazole - 5 - carboxylic acid (20 mg, 0.056 mmol, 1.0 equiv.), (3R,4R) - 3 - (3,4 - dihydroisoquinolin - 2(1H) - yl)piperidin - 4 - ol (13 mg, 0.056 mmol, 1.0 equiv.) and O - (7 - azabenzotriazol - 1 - yl) - N,N,N ',N ' - tetramethyluronium hexafluorophosphate (32 mg, 0.084 mmol, 1.5 equiv.) were added to N,N - dimethylformamide (0.2 mL), and further N,N - diisopropylethylamine (22 mg, 0.168 mmol, 3.0 equiv.) was added. The mixture was stirred at 16 °C for 0.5 h, and the completion of the reaction of the starting materials was detected by TLC. Water (15 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate three times (3 × 15 mL). The organic phases were combined, washed once with a saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered by suction, the solvent was concentrated, and the crude product was separated and purified by Prep - HPLC (C18, 10 mmol / L aqueous solution / acetonitrile) to obtain the target compound (2.06 mg, yield: 6.4%).

[0640] LC - MS (ESI) [M + H] + = 574.5; 1 1H NMR (400 MHz, CD3OD) δ 7.31 - 7.14 (m, 5H), 7.12 - 7.05 (m, 3H), 7.04 - 7.00 (m, 1H), 4.39 - 4.32 (m, 1H), 4.32 - 4.10 (m, 2H), 3.95 (s, 2H), 3.92 - 3.74 (m, 4H), 3.53 - 3.45 (m, 1H), 3.28 - 3.21 (m, 1H), 2.95 - 2.84 (m, 5H), 2.80 - 2.62 (m, 3H), 2.17 - 2.11 (m, 1H), 2.10 (s, 3H), 2.07 - 1.99 (m, 1H), 1.88 - 1.78 (m, 1H), 1.54 - 1.29 (m, 3H).

[0641] Example 390: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(4-bromophenyl)methanone

Chemical Structure

[0642] (3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (500 mg, 2.152 mmol, 1.0 equiv.) was dissolved in dichloromethane (20 mL), and triethylamine (330 mg, 3.229 mmol, 1.5 equiv.) was added. The ice-salt bath was cooled to about -10 °C, and then a solution of p-bromobenzoyl chloride (520 mg, 2.368 mmol, 1.1 equiv.) in dichloromethane (5 mL) was gradually added to the reaction solution. Finally, the temperature was raised to room temperature (20 °C) and stirred for half an hour, and the completion of the reaction was monitored by LC-MS. Water was added to the reaction solution to quench the reaction, and the mixture was extracted with dichloromethane. The combined organic layers were washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product, which was separated and purified by reverse-phase HPLC (C18, aqueous NH4HCO3 / acetonitrile) to obtain the title compound (697.22 mg, yield 77.9%).

[0643] LC-MS (ESI) [M+H] + = 416.0; 1 1H NMR (400 MHz, (CD3)2SO) δ 7.58 - 7.49 (m, 2H), 7.34 - 7.28 (m, 2H), 7.19 - 7.09 (m, 3H), 7.07 - 6.98 (m, 1H), 5.20 - 4.73 (m, 1H), 4.18 - 3.80 (m, 2H), 3.78 - 3.39 (m, 3H), 3.17 - 3.01 (m, 1H), 2.99 - 2.56 (m, 6H), 2.05 - 1.35 (m, 2H).

[0644] Example 391: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-isobutoxy-6-((1-(thiazol-2-yl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone hydrochloride

[0645] Step 1: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-isobutoxy-6-((1-(thiazol-2-yl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone

Chemical formula

[0646] ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-isobutoxy-6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone (1 g, 1.97 mmol, 1.0 equiv.), 2-bromothiazole (387 mg, 2.36 mmol, 1.2 equiv.), CuI (37.44 mg, 0.2 mmol, 0.1 equiv.), L-proline (45.27 mg, 0.4 mmol, 0.2 equiv.) and K2CO3 (543 mg, 3.93 mmol, 2.0 equiv.) were dissolved in DMSO (10 mL) and reacted at 100 °C for 16 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, the organic phase was concentrated, the solvent was dried by rotary evaporator, and the crude product was purified by prep-HPLC (C18, 10 mmol / L aqueous NH4OH / acetonitrile) to obtain the target compound (310 mg, yield 26.6%). LC-MS (ESI) [M + H] + = 592.2。

[0647] Step 2: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-isobutoxy-6-((1-(thiazol-2-yl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone hydrochloride

Chem.

[0648] ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-isobutoxy-6-((1-(thiazol-2-yl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone (310 mg, 0.524 mmol, 1.0 equiv.) and hydrochloric acid (1 M) (0.55 mL, 0.55 mmol, 1.05 equiv.) were dissolved in MeOH (10 mL) and reacted at 20 °C for 0.5 h. The reaction solution was concentrated to obtain the target compound (310.18 mg, yield 95%).

[0649] LC-MS (ESI) [M+H] + = 592.2; 1 H NMR (400 MHz, (CD3)2SO) δ 10.56 - 10.37 (m, 1H), 8.18 (s, 1H), 7.28 - 7.18 (m, 5H), 6.89 - 6.88 (m, 1H), 6.47 - 6.09 (m, 2H), 4.74 - 4.41 (m, 3H), 4.13 (s, 1H), 4.03 - 3.89 (m, 6H), 3.78 - 3.55 (m, 3H), 3.35 - 3.26 (m, 3H), 3.14 - 2.98 (m, 2H), 2.86 - 2.65 (m, 1H), 2.26 - 2.15 (m, 1H), 2.03 - 1.98 (m, 3H), 1.84 - 1.75 (m, 1H), 1.62 - 1.50 (m, 2H), 1.61 - 1.53 (m, 6H).

[0650] Example 392: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-((1-(4-methylbenzoyl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone hydrochloride

[0651] Step 1: Preparation of 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-methoxypyrimidine-4-carboxylic acid

Chemical formula

[0652] Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-4-carboxylate (2.0 g, 5.4 mmol, 1.0 equiv.) and sodium methoxide (1.16 g, 21.6 mmol, 4.0 equiv.) were dissolved in methanol (20 mL), and the reaction mixture was stirred at 80 °C for 16 h. The completion of the reaction was monitored by LC-MS. The reaction solution was adjusted to pH 5 with dilute hydrochloric acid, and the solvent was dried using a rotary evaporator. The crude product was dissolved in methanol, filtered, and the filtrate was dried using a rotary evaporator to obtain 2.5 g of the crude title compound, which was used directly in the next reaction. LC-MS (ESI) [M + H] + = 353.2.

[0653] Step 2: Preparation of tert-butyl 4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-carbonyl)-2-methoxypyrimidin-4-yl)amino)piperidine-1-carboxylate

Chemical formula

[0654] 6-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-methoxypyrimidine-4-carboxylic acid (2.2 g, 6.77 mmol, 1.0 equiv.), (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (1.7 g, 7.45 mmol, 1.1 equiv.), (HATU (3.86 g, 10.1 mmol, 1.5 equiv.) and DIPEA (2.62 g, 20.3 mmol, 3.0 equiv.) were dissolved in DMF (20 mL), and the reaction mixture was stirred at 25 °C for 1 h. The completion of the reaction was monitored by LC-MS. The reaction solution was quenched by adding water, extracted with ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by flash chromatography to obtain the title compound (3.0 g, yield 84%). LC-MS (ESI) [ M + H] + = 567.2。

[0655] Step 3: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone

Chemical Structure

[0656] tert-Butyl 4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-carbonyl)-2-methoxypyrimidin-4-yl)amino)piperidine-1-carboxylate (2.5 g, 4.41 mmol, 1.0 equiv.) was dissolved in methanol (10 mL), and HCl / 1,4-dioxane (5 mL) was added. The reaction mixture was stirred at 20 °C for 1 h. The completion of the reaction was monitored by LC-MS, and the solvent was dried by rotary evaporator to obtain the crude product (2.2 g) of the title compound. LC-MS (ESI) [ M + H] += 467.3。

[0657] Step 4: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-((1-(4-methylbenzoyl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone

Chem.

[0658] ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone (1.0 g, 2.14 mmol, 1.0 equiv.) was dissolved in dichloromethane (10 mL), then triethylamine (1.7 g, 17.1 mmol, 8.0 equiv.) was added. The reaction was cooled to around 0 °C, and then 4-methylbenzoyl chloride (329 mg, 2.14 mmol, 1.0 equiv.) was slowly added to the reaction solution. After stirring for 1 hour, LC-MS indicated the completion of the reaction. After adding water to quench the reaction solution, it was extracted with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, then filtered, concentrated, and the crude product was separated and purified by reverse-phase HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain the title compound (820 mg, yield: 65.5%). LC-MS (ESI) [M + H] + = 585.2。

[0659] Step 5: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-((1-(4-methylbenzoyl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone hydrochloride

Chem.

[0660] ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-((1-(4-methylbenzoyl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone (820 mg, 1.40 mmol, 1.0 equiv.) was dissolved in methanol (10 mL), and then HCl (1 mol / L) (1.47 mL, 1.47 mmol, 1.05 equiv.) was added. The reaction mixture was stirred at 25 °C for 1 h. After the reaction mixture was concentrated, pure water was added to dissolve it, and it was freeze-dried to obtain the title compound (747.47 mg, yield 86%).

[0661] LC-MS (ESI) [M+H] + = 585.2; 1 H NMR (400 MHz, (CD3)2SO) δ 10.58 - 10.47 (m, 1H), 8.10 (s, 1H), 7.29 - 7.20 (m, 8H), 6.49 - 6.14 (m, 2H), 4.70 - 4.32 (m, 4H), 4.13 - 3.98 (m, 3H), 3.84 - 3.75 (m, 4H), 3.56 - 2.65 (m, 9H), 2.39 - 2.17 (m, 4H), 1.92 - 1.81 (m, 3H), 1.43 - 1.42 (m, 2H).

[0662] Example 393: Preparation of ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-((1-(1-methyl-1H-pyrazole-4-carbonyl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone hydrochloride

[0663] Step 1: Preparation of ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-((1-(1-methyl-1H-pyrazole-4-carbonyl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone [Chemical formula]

[0664] ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone (1.0 g, 2.14 mmol, 1.0 equiv.), 1-Methylpyrazole-4-carboxylic acid (297 mg, 2.36 mmol, 1.1 equiv.), HATU (1.22 g, 3.21 mmol, 1.5 equiv.) and DIPEA (831 mg, 6.43 mmol, 3.0 equiv.) were dissolved in DMF (10 mL) and reacted at 20 °C for 1 hour. The reaction mixture was concentrated and the crude product was separated and purified by reverse-phase HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain the title compound (709 mg, yield 57.7%). LC-MS (ESI) [M + H] + = 575.2.

[0665] Step 2: Preparation of ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-((1-(1-methyl-1H-pyrazole-4-carbonyl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone hydrochloride [Chemical formula]

[0666] ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(2-methoxy-6-((1-(1-methyl-1H-pyrazole-4-carbonyl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone (709 mg, 1.23 mmol, 1.0 equiv.) was dissolved in methanol (10 mL), and then HCl (1 mol / L) (1.29 mL, 1.29 mmol, 1.05 equiv.) was added. The reaction mixture was stirred at 25 °C for 1 h. After concentrating the reaction mixture, pure water was added to dissolve it, and then freeze-dried to obtain the title compound (694.42 mg, yield 92.4%).

[0667] LC-MS (ESI) [M+H] + = 575.2; 1 H NMR (400 MHz, (CD3)2SO) δ 10.75 - 10.34 (m, 1H), 8.35 - 8.05 (m, 2H), 7.64 (s, 1H), 7.27 - 7.20 (m, 4H), 6.37 - 6.11 (m, 2H), 4.68 - 4.46 (m, 3H), 4.13 - 3.96 (m, 5H), 3.85 - 3.81 (m, 6H), 3.55 - 2.64 (m, 9H), 2.33 - 2.13 (m, 1H), 1.96 - 1.80 (m, 3H), 1.42 - 1.39 (m, 2H).

[0668] Example 394: Preparation of ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-((1-(5-methylthiazol-2-yl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone hydrochloride

[0669] Step 1: Preparation of 1-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-((1-(5-methylthiazol-2-yl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone

Chemical formula

[0670] ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone (0.52 g, 0.87 mmol, 1.0 equiv.), 2-chloro-5-methylthiazole (139 mg, 1.04 mmol, 1.2 equiv.) and sodium tert-butoxide (418 mg, 4.35 mmol, 5.0 equiv.) were dissolved in DMF (5 mL) and reacted at 90 °C for 40 h under nitrogen protection. The reaction mixture was concentrated, and the crude product was purified by preparative reverse-phase HPLC (C18, 10 mmol / L aqueous NH4HCO3 / acetonitrile) to obtain the target compound (92 mg, yield 19.8%). LC-MS (ESI) [M + H] + = 534.2。

[0671] Step 2: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-((1-(5-methylthiazol-2-yl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone hydrochloride

Chemical formula

[0672] ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-((1-(5-methylthiazol-2-yl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone (92 mg, 0.17 mmol, 1.0 equiv.) and hydrochloric acid (0.1 M) (1.9 mL, 0.19 mmol, 1.1 equiv.) were dissolved in MeOH (3 mL) and reacted at 20 °C for 0.5 h. The reaction solution was concentrated to obtain the target compound (83.06 mg, yield 85.7%).

[0673] LC-MS (ESI) [M-Cl] + = 534.2; 1 H NMR (400 MHz, (CD3)2SO) δ 9.11 - 8.85 (m, 2H), 8.54 - 8.31 (m, 1H), 8.07 - 7.82 (m, 1H), 7.30 - 6.96 (m, 4H), 6.92 - 6.57 (m, 2H), 5.22 - 5.01 (m, 1H), 4.81 - 4.36 (m, 1H), 4.25 - 3.96 (m, 2H), 3.89 - 3.61 (m, 2H), 3.32 - 3.25 (m, 3H), 3.20 - 2.87 (m, 6H), 2.82 - 2.59 (m, 2H), 2.28 - 2.19 (m, 3H), 2.09 - 1.65 (m, 6H).

[0674] Example 395: Preparation of ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-((1-(benzenesulfonyl)piperidin-4-yl)amino)pyrimidin-4-yl)methanone

Chemical formula

[0675] ((3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)(6-(piperidin-4-ylamino)pyrimidin-4-yl)methanone (150 mg, 0.343 mmol, 1.0 equiv.) was dissolved in dichloromethane (5 mL), then triethylamine (42 mg, 0.412 mmol, 1.2 equiv.) was added, and the reaction was cooled to around -10 °C. Then, while maintaining the temperature of the reaction system not exceeding 0 °C, benzenesulfonyl chloride (67 mg, 0.378 mmol, 1.1 equiv.) was slowly added to the reaction solution. After completion, the reaction was gradually warmed to room temperature (20 °C). After stirring for 1 hour, the completion of the reaction was monitored by LC-MS. Water was added to the reaction solution to quench it, and then the reaction solution was concentrated to obtain a crude product, which was separated and purified by reverse-phase HPLC (C18, aqueous NH4HCO3 / acetonitrile) to obtain the title compound (42 mg, yield: 21.2%). LC-MS (ESI) [M + H] + = 577.2。

[0676] Example 396: Preparation of (6-((1-(Cyclohexylcarbonyl)piperidin-4-yl)amino)-2-(cyclopentylthio)pyrimidin-4-yl)((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone

Chemical Structure

[0677] (2-(Cyclopentylthio)-6-(piperidin-4-ylamino)pyrimidin-4-yl)((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone (150 mg, 0.280 mmol, 1.0 equiv.) was dissolved in dichloromethane (5 mL), then triethylamine (86 mg, 0.839 mmol, 3.0 equiv.) was added. The reaction was cooled to around -10 °C, and then cyclohexylcarbonyl chloride (45 mg, 0.308 mmol, 1.1 equiv.) was slowly added to the reaction solution while maintaining the temperature of the reaction system not exceeding 0 °C. After the addition, the reaction was gradually warmed to room temperature (20 °C). After stirring for 1 hour, the completion of the reaction was monitored by LC-MS. Water was added to the reaction solution to quench it, and then the reaction solution was concentrated to obtain a crude product, which was separated and purified by reverse-phase HPLC (C18, aqueous NH4HCO3 / acetonitrile) to obtain the title compound (50 mg, yield: 27.8%). LC-MS (ESI) [ M + H] + = 647.0。

[0678] Example 397: Preparation of (6-(Cyclobutylamino)-2-((1-(dimethylamino)propan-2-yl)oxy)pyrimidin-4-yl)((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone

[0679] Step 1: Preparation of 6-(Cyclobutylamino)-2-((1-(dimethylamino)propan-2-yl)oxy)pyrimidine-4-carboxylic acid

Chemical formula

[0680] Methyl 2-chloro-6-(cyclobutylamino)pyrimidine-4-carboxylate (60 mg, 0.25 mmol) was dissolved in 1-(dimethylamino)propan-2-ol (770 mg, 7.50 mmol), p-toluenesulfonic acid (43 mg, 0.25 mmol) was added at room temperature, the reaction solution was heated to 110 °C, and the reaction was carried out for 16 hours. The completion of the reaction was monitored by TLC. The reaction solution was prepared under medium pressure to obtain the title compound (30 mg, yield: 38.9%). LC-MS (ESI) [ M + H] + = 295.2。

[0681] Step 2: Preparation of (6-(cyclobutylamino)-2-((1-(dimethylamino)propan-2-yl)oxy)pyrimidin-4-yl)((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone

Chemical formula

[0682] 6-(Cyclobutylamino)-2-((1-(dimethylamino)propan-2-yl)oxy)pyrimidine-4-carboxylic acid (30 mg, 0.102 mmol) and triethylamine (31 mg, 0.306 mmol) were dissolved in N,N-dimethylformamide (3 mL), and urea N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate (46 mg, 0.122 mmol) was added at room temperature. The mixture was stirred at room temperature for 10 minutes, (3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (35.5 mg, 0.153 mmol) was added, and after the addition was complete, the mixture was stirred at room temperature for 16 hours. It was confirmed by TLC that the reaction of the raw materials was complete. 20 mL of water and 20 mL of dichloromethane were added to the reaction system, extracted, and the organic layer was separated after standing. The organic layer was washed 3 times with saturated brine and dried over anhydrous sodium sulfate to obtain a crude product, and the title compound was obtained from medium pressure (4.11 mg, yield 8.1%). LC-MS (ESI) [M + H] + = 509.3。

[0683] Example 398: Preparation of 1-(4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl-4’,4’-dideuterium)-3-hydroxypiperidin-1-carbonyl)-2-isobutoxypyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

[0684] Step 1: First Preparation of tert-butyl (3R,4R)-3-((tert-butoxycarbonyl)oxy)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate

Chem.

[0685] (3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (1 g, 4.3 mmol, 1.0 equiv.), triethylamine (1.3 g, 12.9 mmol, 3.0 equiv.) and DMAP (105 mg, 0.86 mmol, 0.2 equiv.) were dissolved in THF (20 mL), and Boc2O (1.88 g, 8.6 mmol, 2.0 equiv.) was added with stirring, and the reaction was carried out at 20 °C for 1 hour. The reaction solution was concentrated and separated and purified by column chromatography (silica gel, PE:EA = 20:1) to obtain the target product (1.12 g, yield 60%). LC-MS (ESI) [M + H] + = 433.3。

[0686] Step 2: Preparation of tert-butyl (3R,4R)-3-((tert-butoxycarbonyl)oxy)-4-(4-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate

Chem.

[0687] tert-Butyl (3R,4R)-3-(((tert-butoxycarbonyl)oxy)-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate (300 mg, 0.49 mmol, 1.0 equiv.) was dissolved in CHCl3 (10 mL), and formic acid (451 mg, 9.8 mmol, 20.0 equiv.) and DDQ (473 mg, 2.08 mmol, 3.0 equiv.) were added. The reaction was carried out at 20 °C for 16 h. Saturated aqueous sodium carbonate solution (5 mL) was added to the reaction mixture, and the mixture was extracted with water and DCM. The organic phase was concentrated, and the crude product was separated and purified by column chromatography (silica gel, DCM:MeOH = 50:1) to obtain the target compound (46 mg, yield 18%). LC-MS (ESI) [M + Na] + = 469.3。

[0688] Step 3: Preparation of 2-(3R,4R-3-hydroxypiperidin-4-yl)-2,3-dihydroisoquinolin-4(1H)-one

Chemical formula

[0689] tert-Butyl (3R,4R)-3-(((tert-butoxycarbonyl)oxy)-4-(4-oxo-3,4-dihydroisoquinolin-2(1H)-yl)piperidine-1-carboxylate (46 mg, 0.1 mmol, 1.0 equiv.) was dissolved in EA (0.5 mL), and ethyl acetate-hydrogen chloride gas (5 mL, 4M) was added. The reaction was carried out at 20 °C for 1 h. The reaction solution was concentrated, and the crude product was dissolved in MeOH:H2O (1 mL:1 mL), and lithium hydroxide (7 mg, 0.3 mmol, 3.0 equiv.) was added. The reaction was carried out at 20 °C for 1 h. The reaction mixture was concentrated and purified by pre-TL (silica gel, DCM:MeOH = 5:1) to obtain the target compound (12 mg, yield 48%). LC-MS (ESI) [M + H] + = 247.3。

[0690] Step 4: Preparation of (3R,4R)-4-(4,4-dideuterio-3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol

Chem.

[0691] 2-((3R,4R)-3-Hydroxypiperidin-4-yl)-2,3-dihydroisoquinolin-4(1H)-one (4 mg, 0.016 mmol, 1.0 equiv.) was dissolved in dry THF (1 mL), LiAlD4 (1.3 mg, 0.032 mmol, 2.0 equiv.) was added, and the reaction was carried out at 20 °C for 2 hours. The reaction solution was quenched with heavy water and concentrated to obtain the title compound (6 mg, crude product). LC-MS (ESI) [M + H] + = 235.1。

[0692] Step 5: Preparation of 1-(4-((6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl-4’,4’-dideuterio)-3-hydroxypiperidin-1-carbonyl)-2-isobutoxypyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chem.

[0693] The title compound was synthesized using the method of Step 3 of Example 187. LC-MS (ESI) [M + H] + = 553.3。

[0694] Example 399: Preparation of 1-(4-((6-((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl-4’,4’-dideuterio)-3-hydroxypiperidine-1-carbonyl)-2-(pentan-3-yloxy)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one

Chem.

[0695] Using the intermediate and method of the second step of Example 188, the title compound was synthesized. LC-MS (ESI) [M + H] + = 567.4.

[0696] Example 400: Preparation of (6-((1-(cyclobutylcarbonyl)piperidin-4-yl)amino)-2-isopropoxypyrimidin-4-yl)((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl-4,4-d2)-3-hydroxypiperidin-1-yl)methanone

Chem.

[0697] The intermediate 3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol was replaced with 3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl-4,4-d2)piperidin-3-ol, and the title compound was synthesized using the same method as in Example 189. LC-MS (ESI) [M + H] + = 579.4.

[0698] Example 401: Preparation of (6-((1-benzoyl)piperidin-4-yl)amino)-2-isopropoxypyrimidin-4-yl)((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl-4,4-d2)-3-hydroxypiperidin-1-yl)methanone [Chemical formula]

[0699] The intermediate 3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl)piperidin-3-ol was replaced with 3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl-4,4-d2)piperidin-3-ol, and the title compound was synthesized using the same method as in Example 183. LC-MS (ESI) [M + H] + = 601.3.

[0700] Example 402: Preparation of (1-(4-((2-(tert-butylsulfanyl)-6-(3R,4R-4-(3,4-dihydroisoquinolin-2(1H)-yl-4,4-d2)-3-hydroxypiperidin-1-carbonyl)pyrimidin-4-yl)amino)piperidin-1-yl)ethan-1-one [Chemical formula]

[0701] Using the intermediate of Production Example 38, the title compound was synthesized by the method of the third step of Example 187. LC-MS (ESI) [M + H] + = 569.3.

[0702] Comparative Example 1: According to the preparation method of Example 13 of WO2020182018A1, the compound of Comparative Example 1 (1.67 g, yield: 50.0%) was obtained. [Chemical formula]

[0703] LC-MS (ESI) [M+H] + = 477.4; 11H NMR (400 MHz, CDCl3) δ 7.12 (dd, J = 7.9, 4.6 Hz, 3H), 7.02 (d, J = 5.7 Hz, 1H), 6.36 (s, 1H), 5.19 (d, J = 7.7 Hz, 1H), 4.72 (d, J = 13.4 Hz, 1H), 4.51 (d, J = 13.3 Hz, 1H), 3.99 (d, J = 13.5 Hz, 1H), 3.84 (s, 2H), 3.79 (s, 1H), 3.21 (t, J = 11.8 Hz, 1H), 3.10 (t, J = 12.1 Hz, 1H), 2.91 (d, J = 4.1 Hz, 3H), 2.80 (d, J = 10.8 Hz, 3H), 2.50 (s, 3H), 2.11 (s, 3H), 2.05 (d, J = 13.7 Hz, 3H), 2.00 (s, 1H), 1.93 (d, J = 11.9 Hz, 1H), 1.70 (dd, J = 23.6, 11.7 Hz, 2H), 1.39 (d, J = 11.3 Hz, 2H).

[0704] Comparative Example 2 (Example 403): Preparation of 1-(4-((6-(3R,4R-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-carbonyl)-2-methylpyrimidin-4-yl)amino)piperidin-1-yl)ethanone

[0705] First Step: Preparation of 6-Hydroxy-2-methylpyrimidine-4-carboxylic acid [Chemical Structure Diagram]

[0706] (Z)-1,4-Diethoxy-1,4-dioxobuta-2-ene-2-olate (30 g, 142.75 mmol, 1 equiv.) was dissolved in water (50 ml), and an aqueous sodium hydroxide solution (23.6 ml, 6 mol / L) was added dropwise at room temperature. The mixture was stirred at 25 °C for 1 hour. After acetimidamide (18.2 g, 314.05 mmol, 2.2 equiv.) was dissolved in water (40 ml), it was added to the reaction system. The pH was adjusted to 11 with an aqueous sodium hydroxide solution (6 mol / L) in an ice bath, and stirring was continued at this temperature for 40 minutes. The pH was adjusted to 1 with an aqueous hydrochloric acid solution (12 mol / L), and suction filtration was carried out under reduced pressure. The filter cake was washed with an aqueous hydrochloric acid solution (2 × 50 ml, 0.1 mol / L) and dried to obtain the target compound (9.5 g, yield 43.2%). LC-MS (ESI) [M + H] + = 155.2。

[0707] Step 2: Preparation of 6-chloro-2-methylpyrimidine-4-carbonyl chloride

Chemical formula

[0708] 6-Hydroxy-2-methylpyrimidine-4-carboxylic acid (1.5 g, 9.73 mmol, 1.0 equiv.) was added to phosphorus oxychloride (20 ml), and the temperature was raised to 110 °C. The reaction was carried out for 1 hour until the solid was completely dissolved. The reaction was completed by LC-MS, cooled to room temperature, and the reaction solution was concentrated under reduced pressure to obtain a crude product, which was used directly in the next reaction. LC-MS (ESI) [M + H] + = 183.1。

[0709] Step 3: The third step of (6-chloro-2-methylpyrimidin-4-yl) ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone

Chemical formula

[0710] (3R,4R)-4-(3,4-Dihydroisoquinolin-2(1H)-yl)piperidin-3-ol (2.19 g, 9.42 mmol, 1.2 equiv.) and DIEA (8.12 g, 62.8 mmol, 8 equiv.) were dissolved in dichloromethane (30 mL), and 6-chloro-2-methylpyrimidine-4-carbonyl chloride (1.5 g, 7.85 mmol, 1 equiv.) (previously dissolved in dichloromethane) was slowly added thereto under an ice bath. The mixture was stirred at 25 °C for 2 hours, water (50 mL) was added to the reaction system, and the mixture was extracted and separated. The organic phase was concentrated, and the residue was purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain the target compound (230 mg, yield: 7.5%). LC-MS (ESI) [M + H] + = 387.2。

[0711] Step 4: Preparation of 1-(4-((6-(3R,4R-4-(3,4-Dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidine-1-carbonyl)-2-methylpyrimidin-4-yl)amino)piperidin-1-yl)ethanone

Chemical formula

[0712] (6-Chloro-2-methylpyrimidin-4-yl) ((3R,4R)-4-(3,4-dihydroisoquinolin-2(1H)-yl)-3-hydroxypiperidin-1-yl)methanone (230 mg, 0.59 mmol, 1.0 equiv.), (4-aminopiperidin-1-yl)ethan-1-one (170 mg, 1.196 mmol, 2.0 equiv.) and triethylamine (421.1 mg, 4.16 mmol, 7.0 equiv.) were dissolved in isopropanol (10 mL), then heated to 90 °C and reacted for 2 hours. The reaction was monitored by LC-MS, and the reaction product was extracted with ethyl acetate and water. The organic phases were combined, dried, concentrated, and purified by preparation to obtain the target compound (83.7 mg, yield: 28.6%).

[0713] LC-MS (ESI) [M+H] + = 493.4; 1 H NMR (400 MHz, CD3OD) δ 8.44 (s, 1H), 7.11 (d, J = 18.3 Hz, 5H), 6.36 (s, 1H), 4.57 (d, J = 12.5 Hz, 1H), 4.40 (d, J = 12.3 Hz, 1H), 4.09 (d, J = 14.8 Hz, 2H), 3.94 - 3.70 (m, 4H), 3.24 (s, 1H), 3.15 - 3.05 (m, 2H), 2.98 (s, 4H), 2.93 - 2.84 (m, 2H), 2.76 - 2.64 (m, 1H), 2.44 (d, J = 8.4 Hz, 3H), 2.10 (s, 3H), 1.98 (dd, J = 35.5, 21.9 Hz, 3H), 1.79 - 1.69 (m, 1H), 1.43 (dd, J = 30.9, 12.2 Hz, 2H).

[0714] Biological test evaluation Hereinafter, the present invention will be further described with reference to examples, but the present invention is not limited to these examples.

[0715] Test Example 1: Evaluation of PRMT5 enzyme activity inhibitory activity 1. Test method: 1× enzyme reaction buffer (10 mM Tris 8.0 (Sigma, Cat. No. T2694-1L), 0.01% Tween-20 (Sigma, Cat. No. P2287-100ML), 1 mM DTT (Sigma, Cat. No. D0632-10G)) was prepared. PRMT5 (Active Motif, Cat. No. 31921) and [3H]-SAM (PerkinElmer, Cat. No. NET155V001MC) were added to the 1× enzyme reaction buffer and adjusted to a 25 / 15-fold mixed solution (final concentration of PRMT5 5 nM, final concentration of [3H]-SAM 0.3 μM). 15 μL of this solution was transferred to a 384-well microplate (Corning 384-well Polypropylene Storage Microplates, Cat. No. 3657) containing different concentrations of the compound (final concentration of DMSO 1%) and incubated at room temperature for 60 minutes. The polypeptide substrate GL-27 (Ac-SGRGKGGKGLGKGGAKRHRKVGG-K)(Biotin) (GL Biochem, Cat. No. 342095) was added to the 1× enzyme reaction buffer to prepare a 25 / 10-fold substrate solution. Then, 10 μL of the polypeptide substrate solution (final concentration of polypeptide substrate 100 nM) was added. After reacting at room temperature for 120 minutes, 5 μL of a 6-fold ice-cold SAM (Sigma, Cat. No. A7007-100MG) solution was added to stop the reaction (final concentration of SAM: 0.125 mM). 25 μL was transferred from the reaction system to a FlashPlate (Streptavidin FlashPlate HTS PLUS, High Capacity, 384-well, PerkinElmer, Cat. No. SMP410A001PK), incubated at room temperature for 1 hour, and the plate was washed 3 times with distilled water containing 0.1% Tween-20. Then, the microplate was read for CPM data (Counts Per Minute) with a MicroBeta instrument.After obtaining the CPM raw data with different compound concentrations, the data was standardized according to the formula Inh% = (Max - Sample) / (Max - Min)*100% to obtain the enzyme activity inhibition rate Inh% at each concentration point (where Max is the CPM value of the positive well containing the enzyme, Min is the CPM value of the negative well without the enzyme, and Sample is the CPM value of the compound-treated sample well). The inhibition rate Inh% (Y) corresponding to each concentration (X) was input into EXCEL, and using the XLfit plugin, the half-inhibitory concentration IC 50 of each compound was calculated from the built-in four-parameter fitting formula Y = Bottom + (Top - Bottom) / (1 + (IC 50 / X)*HillSlope).

[0716] 2. Test results: The compounds of the examples shown in the present invention showed the following biological activities in the PRMT5 enzyme activity inhibition test as shown in Table 1 below.

Table 8-1

Table 8-2

[0717] 3. Test conclusions: As shown in the above table, the compounds of the present invention showed different PRMT5 enzyme activity inhibition activities. The measured IC 50 values of some compounds reached the D level (for example, the compound of Example 37 was 99.80 nM, the compound of Example 59 was 107.00 nM, and the compound of Example 73 was 110.00 nM). The measured IC 50 values of some compounds reached the C level (for example, the compound of Example 1 was 65.00 nM, the compound of Example 70 was 66.40 nM, and the compound of Example 173 was 51.25 nM). The measured IC 50 values of some compounds reached the B level (for example, the compound of Example 12 was 35.20 nM, the compound of Example 46 was 29.15 nM, and the compound of Example 172 was 20.54 nM). The measured IC 50The measured values reached the A level (for example, the compound of Example 153 was 6.97 nM, the compound of Example 183 was 6.40 nM, the compound of Example 193 was 6.40 nM, the compound of Example 194 was 4.50 nM, the compound of Example 202 was 5.20 nM, the compound of Example 207 was 3.90 nM, the compound of Example 213 was 4.50 nM, the compound of Example 315 was 4.70 nM, the compound of Example 392 was 2.80 nM, the compound of Example 401 was 7.90 nM), and showed a D level. The compound of Comparative Example 1 (that is, WO2020182018A1 Compound 13, with an IC 50 measured value of 115.00 nM) showed significantly superior effects or comparable effects when compared with any of them.

[0718] Also, the compound of Comparative Example 2 reached a B level in terms of the enzyme inhibition IC 50 measured value (IC 50 measured value of 36.00 nM). The presence of a hydroxyl group substitution at a specific site of the piperidine ring and the presence of a 3,4-trans-disubstituted configuration were shown to be key points for the compounds of the present invention to exhibit excellent enzyme activity inhibition activity.

[0719] Test Example 2: Evaluation of the cell growth inhibitory activity against human B-cell non-Hodgkin lymphoma Z-138 1. Samples and equipment 1) Cell line and culture method

Table 9

[0720] 2) Medium and reagents

Table 10

[0721] 3) 384-well microplate Cornin (registered trademark) 384-well microplate, transparent flat-bottom white polystyrene microwell plate (with cover, sterile), Corning, product number 3765.

[0722] 4) Instruments 2104 EnVision Plate Reader, PerkinElmer, Vi-Cell XR Cell Counter, beckmancoulter.

[0723] 2. Experimental Methods and Procedures 1) Cell Culture The cells were revived and cultured in an incubator at 37°C and 5% CO2 under the culture conditions shown in the above table. They were passaged regularly, and a cell line with good growth state was selected after about 2 generations and used for plating.

[0724] 2) Plating I. The cells were taken out from the culture flask, the cell suspension was transferred to a 50 mL centrifuge tube, centrifuged at 800 - 1000 rpm for 3 - 5 minutes, and then the supernatant was discarded. An appropriate volume of medium was added to the centrifuge tube, and the cells were gently pipetted to resuspend them uniformly. They were counted using a Vi-Cell XR Cell Counter. II. The cell suspension was adjusted to an appropriate concentration according to the measured cell density. III. The cell suspension was added to a 384-well microplate at 40 μL / well, containing 700 cells / well, and an equal volume of cell-free culture was added to the blank control wells.

[0725] 3) Compound Preparation and Addition I. The compound was dissolved in 100% DMSO to prepare a stock solution with a concentration of 10 mM. II. 10 mM concentration of the stock solution was taken, diluted with DMSO to a 2 mM solution, and this was used as the starting concentration and diluted 4-fold in 9 steps with DMSO. III. 200 nL of the gradient concentration solution of the above compound was taken separately and added to each well. 200 nL of DMSO was added to the blank control wells and DMSO control wells so that the final concentration of DMSO was 0.5%. IV. The cell plate was incubated in a carbon dioxide incubator for 5 days (120H).

[0726] 4) Reagent Preparation and Detection Detection was performed according to the instruction manual of the Promega CellTiter-Glo Luminescent Cell Viability Assay Kit (Promega-G7573): I. The CellTiter-Glo buffer was dissolved and left at room temperature. II. The lyophilized CellTiter-Glo substrate was left at room temperature. III. The CellTiter-Glo buffer was added to one vial of the CellTiter-Glo substrate to dissolve the substrate and prepare the CellTiter-Glo working solution. IV. It was fully dissolved by gentle shaking. V. The cell culture plate was taken out and left to equilibrate at room temperature. VI. 25 μL of the CellTiter Glo reagent was uniformly added to each well, shaken for 10 minutes while protecting from light, and incubated for 10 minutes. VII. The luminescence signal was detected using a 2104 EnVision plate reader.

[0727] 3. Data Analysis The inhibition rate (IR) of the test compound was calculated using the following formula: IR (%) = (1 - (RLU 化合物 - RLU ブランク対照 ) / (RLU DMSO - RLU ブランク対照 )) * 100%, The drug efficacy inhibition rate curve was plotted using XLFit, and the IC 50 value was calculated using the following four-parameter model: [fit=(A + ((B - A) / (1 + ((C / x)^D))))]

[0728] 4. Experimental Results: The compounds of the examples shown in the present invention showed the biological activities in Table 2 below in the Z-138 cell growth inhibition test.

Table 11-1

Table 11-2

[0729] 5. Test Conclusion: As shown in the above table, the compounds of the present invention exhibit different Z-138 cell growth inhibitory activities, and the cell growth inhibitory IC 50 The measured values reach the E level (for example, the compound of Example 45 is 1325.00 nM, the compound of Example 66 is 1358.00 nM, and the compound of Example 259 is 1190.00 nM), and the cell growth inhibitory IC 50 The measured values reach the D level (for example, the compound of Example 39 is 134.70 nM, the compound of Example 215 is 157.00 nM, and the compound of Example 276 is 111.00 nM), and the cell growth inhibitory IC 50 The measured values reach the C level (for example, the compound of Example 170 is 62.80 nM, the compound of Example 217 is 60.50 nM, and the compound of Example 339 is 65.30 nM), and the cell growth inhibitory IC 50 The measured values reach the B level (for example, the compound of Example 165 is 33.70 nM, the compound of Example 183 is 46.60 nM, and the compound of Example 212 is 22.22 nM), and the cell growth inhibitory IC 50 The measured values reach the A level (for example, the compound of Example 153 is 13.75 nM, the compound of Example 158 is 19.00 nM, the compound of Example 171 is 11.50 nM, the compound of Example 194 is 11.00 nM, the compound of Example 202 is 8.00 nM, the compound of Example 207 is 12.00 nM, the compound of Example 243 is 5.80 nM, the compound of Example 253 is 7.40 nM, the compound of Example 392 is 12.30 nM, the compound of Example 401 is 16.00 nM). When compared with the compound of Comparative Example 1 showing the E level (i.e., WO2020182018A1 compound 13 with an IC 50 measured...

Claims

1. A compound represented by formula (III), or a pharmaceutically acceptable salt thereof. 【Chemical 1】 In the formula, R 16 is selected from hydrogen, deuterium, halogen, hydroxyl, mercapto, amino, cyano, -R 9 , -OR 9 , -SR 9 , -NH(R 9 ), and -N(R 9 )(R 10 ), and R 9 and R 10 are each independently selected from C 1-6 alkyl and C 3-6 cycloalkyl, or R 9 and R 10 together with the N atom to which they are attached form a 4- to 6-membered heterocyclic group, R 11 is C 6-8 selected from aryl and 5- to 6-membered heteroaryl, wherein the aryl or heteroaryl is optionally substituted with one or more of halogen, C 1-6 alkyl, and trifluoromethyl The heterocyclic group or heteroaryl group each independently contains 1, 2 or 3 heteroatoms selected from N, O and S.

2. R 16 is hydrogen, deuterium, -R 9 , -OR 9 , -SR 9 , and -N(R 9 )(R 10 ), provided that R 9 and R 10 are as defined in claim 1, a compound according to claim 1, or a pharmaceutically acceptable salt thereof.

3. R 16 is selected from hydrogen, deuterium, -OCH 3 -, -OCH 2 CH 3 -, -OCH(CH 3 ) 2 -, -OCH 2 CH(CH 3 ) 2 -, -OC(CH 3 ) 3 -, -OCH(CH 2 CH 3 ) 2 -, -O-cyclopropyl, -O-cyclobutyl, -O-cyclopentyl, -O-cyclohexyl, -SCH 3 -, -SCH 2 CH 3 -, -SCH(CH 3 ) 2 -, -SCH 2 CH(CH 3 ) 2 -, -SC(CH 3 ) 3 -, -SCH(CH 2 CH 3 ) 2 -, -S-cyclopropyl, -S-cyclobutyl, -S-cyclopentyl, -S-cyclohexyl, azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, and 3-azabicyclo[3.1.0]hex-3-yl The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

4. R 11 is selected from phenyl, pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyridazinyl, and pyrazinyl, and the phenyl, pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyridazinyl, or pyrazinyl is optionally substituted with one or more of fluorine, methyl, and trifluoromethyl, the compound according to claim 1, or a pharmaceutically acceptable salt thereof.

5. R 11 is selected from phenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-methylphenyl, 3-methylphenyl, 4-methylphenyl, 2-(trifluoromethyl)phenyl, 3-(trifluoromethyl)phenyl, 4-(trifluoromethyl)phenyl, thiophen-2-yl, thiophen-3-yl, 1-methyl-1H-pyrazol-3-yl, 1-methyl-1H-pyrazol-4-yl, 1-methyl-1H-pyrazol-5-yl, isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl, thiazol-2-yl, thiazol-4-yl, thiazol-5-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, and pyrimidin-5-yl The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

6. The compound represented by formula (III) is a compound represented by formula (III)A or formula (III)B, 【Chemical 2】 wherein R 11 and R 16 are as defined in claim 1 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

7. The following compound, or a pharmaceutically acceptable salt thereof. 【Table 1-1】 【Table 1-2】 【Table 1-3】 【Table 1-4】 【Table 1-5】 【Table 1-6】 【Table 1-7】 【Table 1-8】 【Table 1-9】 【Table 1-10】 【Table 1-11】 【Table 1-12】 【Table 1-13】 【Table 1-14】 【Table 1-15】 【Table 1-16】 【Table 1-17】 【Table 1-18】 【Table 1-19】

8. A pharmaceutical composition comprising the compound according to any one of claims 1-7, or a pharmaceutically acceptable salt thereof.

9. Use of the compound according to any one of claims 1-7, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 8, in the preparation of a medicament for the prevention and / or treatment of a disease mediated by PRMT5.

10. The use according to claim 9, wherein the disease mediated by PRMT5 is a cancer or tumor-related disease.

11. The use according to claim 9, wherein the disease mediated by PRMT5 is lymphoma.

12. The compound according to any one of claims 1-7, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 8, for the prevention and / or treatment of a disease mediated by PRMT5.

13. The compound or pharmaceutical composition according to claim 12, wherein the disease mediated by PRMT5 is a cancer or tumor-related disease.

14. The compound or pharmaceutical composition according to claim 12, wherein the disease mediated by PRMT5 is lymphoma.

15. A combined pharmaceutical composition comprising the compound according to any one of claims 1-7, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 8, and other anticancer agents or immune checkpoint inhibitors.

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