A 1,3-disubstituted tetrahydropyrrole derivative and its preparation method and application

Abstract

The present invention belongs to the field of pharmaceutical technology, and specifically relates to a 1,3-disubstituted tetrahydropyrrole derivative, a preparation method and an application thereof. The present invention provides a 1,3-disubstituted tetrahydropyrrole derivative, a preparation method and an application thereof, and characterizes the in vitro anti-tumor cell activity and in vitro enzyme inhibition activity of the 1,3-disubstituted tetrahydropyrrole derivative at the cellular level. The 1,3-disubstituted tetrahydropyrrole derivative provided by the present invention belongs to a non-adenosine inhibitor, has the advantages of high metabolic stability, novel structure, and simple and easy preparation method.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to a 1,3-disubstituted tetrahydropyrrole derivative and a preparation method and application thereof. Background Art

[0002] DOT1L (Disruptor of telomeric silencing 1-like) is the only known histone H3 lysine 79 (H3K79) methyltransferase and the only member of the histone methyltransferase family that does not contain a SET domain. It was first reported in 2002. DOT1L plays a key role in transcriptional regulation, DNA repair, embryonic development, and cell cycle regulation. Inhibiting its expression will promote tumor cell differentiation and apoptosis. Therefore, DOT1L has become an important target for tumor treatment.

[0003] In recent years, studies have found that there are many pathological mutations in the functional activity domain of DOT1L protein, among which the gain-of-function mutation R231Q is the most representative, such as the technology disclosed in Chinese patent CN114214407. Based on a large number of experiments, it has been confirmed that the R231Q mutation can improve the binding ability of DOT1L protein with substrates and enhance its own activity, thereby showing a stronger effect in promoting tumor cell proliferation, self-renewal, invasion and metastasis, and drug resistance. However, the current research on DOT1L R231Q There are relatively few effective inhibitors, which is due to the interaction with the wild-type protein (DOT1L WT ) compared to the mutated DOT1L R231Q The protein has a more flexible and larger ligand binding cavity, which makes the existing DOT1L WT Inhibitors of DOT1L R231Q The inhibitory effect weakens or disappears.

[0004] In addition, DOT1L currently developed at home and abroad WT Most inhibitors belong to adenosine compounds, and their molecular structures contain adenosine or adenosine-like nuclei (e.g., 7-azapurine compounds, 2'-C-methyl substituted nucleoside compounds), such as the technologies disclosed in invention patents WO2012075500, WO2012075381, CN109748943, and CN110092804. However, adenosine compounds contain metabolically unstable aminal fragments in their nucleus structures, which are easily recognized and rapidly cleaved by a variety of metabolic enzymes (e.g., adenosine deaminase, hydrolase, or nucleosidase) in the human body (Khirsariya P, P, Maier L, et al., Journal of Medicinal Chemistry, 2022, 65: 5701-5723.), therefore showing the disadvantages of high clearance rate and poor pharmacokinetic properties. At present, the research and development of DOT1L inhibitors is still in its infancy, and no drugs have been approved for marketing. Therefore, it is necessary to develop a non-adenosine DOT1L inhibitor with good pharmacokinetic properties.

[0005] Pyrrole and its derivatives are non-adenosine compounds with diverse biological activities. Studies have shown that pyrrole derivatives have significant inhibitory activity against various types of tumors, and when pyrrole derivatives are combined with other anti-tumor drugs for treatment, they can improve the efficacy and reduce the occurrence of drug resistance, which has attracted the attention of pharmaceutical researchers and is a potential anti-tumor drug structure type. There are currently no reports of pyrrole derivatives as DOT1L inhibitors. Summary of the invention

[0006] In view of the problems existing in the background technology, the present invention provides a 1,3-disubstituted tetrahydropyrrole derivative belonging to a non-adenosine inhibitor.

[0007] The object of the present invention is to provide a 1,3-disubstituted tetrahydropyrrole derivative, the chemical structure of which is shown in the general formula (I) or (II):

[0008]

[0009] Where:

[0010] Het is 0-4 independent R 1 substituted 8-10 membered heteroaryl containing 1-4 heteroatoms selected from N, O and S;

[0011] R 1 is halogen, hydroxy, amino, nitro, cyano, trifluoromethyl, trifluoromethoxy, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkoxy, (C 1 -C 6 ) acylamino or 1-2 (C 1 -C 6 ) alkyl-substituted amino;

[0012] R 2 For hydrogen, (C 1 -C 6 ) alkyl or (C 1 -C 6 ) acyl group;

[0013] R3 For hydrogen, (C 1 -C 6 ) alkyl, (C 2 -C 6 ) alkenyl, (C 2 -C 6 ) alkynyl, halogen or amino or cyano or carboxyl substituted (C 1 -C 6 )alkyl, (CH 2 ) p (C 3 -C 6 )cycloalkyl, (C 1 -C 6 ) acyl, (C 1 -C 6 )sulfonyl, (CH 2 ) p NR 5 R 6 , (CH 2 ) p CONR 5 R 6 , (CH 2 ) p COOR 5 or (CH 2 ) p OR 5 ;

[0014] R 5 and R 6 are the same or different and are independently selected from hydrogen, (C 1 -C 6 ) alkyl and halogen or amino or hydroxy or cyano substituted (C 1 -C 6 ) alkyl; or R 5 and R 6 Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic or heteroaryl group, wherein the heterocyclic or heteroaryl group, in addition to R 5 and R 6 In addition to the nitrogen atom connected, it contains 0-4 heteroatoms selected from N, O and S;

[0015] X is methylene, (CH 2 ) m CONH(CH 2 ) n , (CH 2 ) m NH(CH 2 ) n , (CH 2 ) m COO(CH 2 )n or (CH 2 ) m O(CH 2 ) n ;

[0016] Ar is a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and S;

[0017] R 4 represents 0-4 substituents, independently selected from halogen, hydroxy, amino, nitro, cyano, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkoxy, (C 3 -C 6 )cycloalkyl, (C 1 -C 6 ) alkoxycarbonyl, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) alkyl, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) alkoxy, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) acyl, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) acylamino and 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 )sulfonylamino;

[0018] p is independently 0-6;

[0019] m is independently 0-3;

[0020] n is independently 0-3.

[0021] Preferably, in the general formula (I) or (II):

[0022] Het is 0-4 independent R 1 substituted 8-10 membered heteroaryl containing 1-4 heteroatoms selected from N, O and S;

[0023] R 1 is halogen, hydroxy, amino, nitro, cyano, trifluoromethyl, trifluoromethoxy, (C 1 -C 3 ) alkyl, (C 1 -C 3 ) alkoxy, (C 1 -C 3) acylamino or 1-2 (C 1 -C 3 ) alkyl-substituted amino;

[0024] R 2 For hydrogen, (C 1 -C 3 ) alkyl or (C 1 -C 3 ) acyl group;

[0025] R 3 For hydrogen, (C 1 -C 3 ) alkyl, (C 2 -C 4 ) alkenyl, (C 2 -C 4 ) alkynyl, halogen or amino or cyano or carboxyl substituted (C 1 -C 3 )alkyl, (CH 2 ) p (C 3 -C 6 )cycloalkyl, (C 1 -C 3 ) acyl, (C 1 -C 3 )sulfonyl, (CH 2 ) p NR 5 R 6 , (CH 2 ) p CONR 5 R 6 , (CH 2 ) p COOR 5 or (CH 2 ) p OR 5 ;

[0026] R 5 and R 6 are the same or different and are independently selected from hydrogen, (C 1 -C 3 ) alkyl and halogen or amino or hydroxy or cyano substituted (C 1 -C 3 ) alkyl; or R 5 and R 6 Together with the nitrogen atom to which it is attached, it forms a 4-6 membered heterocyclic or heteroaryl group, wherein the heterocyclic or heteroaryl group, in addition to R 5 and R 6 In addition to the nitrogen atom connected, it contains 0-4 heteroatoms selected from N, O and S;

[0027] X is methylene, (CH 2 ) m CONH(CH 2 ) n , (CH 2 ) m NH(CH 2 ) n , (CH 2 ) m COO(CH 2 ) n or (CH 2 ) m O(CH 2 ) n ;

[0028] Ar is a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and S;

[0029] R 4 represents 0-3 substituents, independently selected from halogen, hydroxy, amino, nitro, cyano, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkoxy, (C 3 -C 6 )cycloalkyl, (C 1 -C 6 ) alkoxycarbonyl, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) alkyl, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) alkoxy, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) acyl, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) acylamino and 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 )sulfonylamino;

[0030] p is independently 0-4;

[0031] m is independently 0-2;

[0032] n is independently 0-2.

[0033] More preferably, in the general formula (I) or (II):

[0034] Het is 0-2 independent R 1substituted 8-10 membered heteroaryl containing 1-4 heteroatoms selected from N, O and S;

[0035] R 1 is halogen, hydroxy, amino, nitro, cyano, trifluoromethyl, trifluoromethoxy, (C 1 -C 3 ) alkyl, (C 1 -C 3 ) acylamino or 1-2 (C 1 -C 3 ) alkyl-substituted amino;

[0036] R 2 For hydrogen, (C 1 -C 3 ) alkyl or (C 1 -C 3 ) acyl group;

[0037] R 3 For hydrogen, (C 1 -C 3 ) alkyl, halogen or amino or cyano or carboxyl substituted (C 1 -C 3 ) alkyl, (C 1 -C 3 ) acyl, (CH 2 ) p NR 5 R 6 , (CH 2 ) p CONR 5 R 6 , (CH 2 ) p COOR 5 or (CH 2 ) p OR 5 ;

[0038] R 5 and R 6 are the same or different, independently selected from hydrogen and (C 1 -C 3 ) alkyl; or R 5 and R 6 Together with the nitrogen atom to which it is attached, it forms a 4-10 membered heterocyclic or heteroaryl group, wherein the heterocyclic or heteroaryl group, in addition to R 5 and R 6 In addition to the nitrogen atom connected, it contains 0-4 heteroatoms selected from N, O and S;

[0039] X is methylene, (CH 2 ) m CONH(CH2 ) n or (CH 2 ) m NH(CH 2 ) n ;

[0040] Ar is a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and;

[0041] R 4 represents 0-2 substituents, independently selected from halogen, hydroxy, amino, nitro, cyano, (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkoxy, (C 1 -C 6 ) alkoxycarbonyl, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) alkyl, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) alkoxy, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) acyl, 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) acylamino and 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 )sulfonylamino;

[0042] p is independently 0-4;

[0043] m is independently 0-2;

[0044] n is independently 0-2.

[0045] Further preferably, in the general formula (I) or (II):

[0046] Het is 0-2 independent R 1 Substituted 9-membered heteroaryl containing 1-4 heteroatoms selected from N, O and S;

[0047] R 1 is halogen, hydroxy, amino, nitro, cyano, trifluoromethyl, trifluoromethoxy or (C 1 -C 3 )alkyl;

[0048] R 2 is hydrogen or (C 1 -C 3 ) acyl group;

[0049] R 3 For hydrogen, (C 1 -C 3 )alkyl, (CH 2 ) p NR 5 R 6 , (CH 2 ) p CONR 5 R or (CH 2 ) p COOR 5 ;

[0050] R 5 and R 6 are the same or different, independently selected from hydrogen and (C 1 -C 3 ) alkyl; or R 5 and R 6 Together with the nitrogen atom to which it is connected, it forms a 4-6 membered heterocyclic group, wherein the heterocyclic group is 5 and R 6 In addition to the nitrogen atom connected, it contains 0-4 heteroatoms selected from N, O and S;

[0051] X is methylene or (CH 2 ) m CONH(CH 2 ) n ;

[0052] Ar is a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and S;

[0053] R 4 represents 0-1 substituents, each independently selected from (C 1 -C 6 ) alkyl, (C 1 -C 6 ) alkoxycarbonyl and 0-3 halogen or hydroxy or amino substituted (C 1 -C 6 ) acylamino group;

[0054] p is independently 0-4;

[0055] m is independently 0-1;

[0056] n is independently 0-1.

[0057] Further preferably, in the general formula (I) or (II):

[0058] Het is 0-2 R 1substituted 9-membered heteroaryl, the heteroaryl being optionally pyrazolo[3,4-d]pyrimidine, imidazo[1,2-a]pyrazine, pyrrolo[2,3-d]pyrimidine or purine;

[0059] R 1 is halogen, amino or methyl;

[0060] R 2 is hydrogen or acetyl;

[0061] R 3 For hydrogen,

[0062] X is

[0063] Ar is a benzene ring, piperidine or thiophene;

[0064] R 4 represents 0-1 substituents independently selected from tert-butyl, methoxycarbonyl and pivaloylamino.

[0065] The 1,3-disubstituted tetrahydropyrrole derivative is specifically selected from the following compounds:

[0066] 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(isopropyl)amino}propyl}urea;

[0067] 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(isopropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea;

[0068] 1-[4-(tert-butyl)phenyl]-3-{3-{{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}(3-morpholinopropyl)amino}propyl}urea;

[0069] 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholinopropyl)amino}propyl}urea;

[0070] 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholinopropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea;

[0071] 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholino-3-oxopropyl)amino}propyl}urea;

[0072] 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholino-3-oxopropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea;

[0073] 1-[4-(tert-butyl)phenyl]-3-{3-{{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}(3-morpholino-3-oxopropyl)amino}propyl}urea;

[0074] 1-[4-(tert-butyl)phenyl]-3-{3-{{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}urea;

[0075] 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}urea;

[0076] 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}urea;

[0077] 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea;

[0078] Methyl 3-{{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}amino}propanoate;

[0079] Methyl 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}amino}propanoate;

[0080] Methyl 3-{{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanoate;

[0081] Methyl 3-{{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanoate;

[0082] Methyl 3-{{[3-acetoxy-1-(2-chloro-9H-purin-6-yl)pyrrolidin-3-yl]methyl}{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}amino}propanoate;

[0083] N-{2-{{4-{{[3-hydroxy-1-(9H-purin-6-yl)]pyrrolidin-3-yl}methyl(methyl)amino}piperidin-1-yl}methyl}-1H-benzo[d]imidazol-5-yl}pivalamide;

[0084] N-{2-{{4-{{[3-hydroxy-1-(2-chloro-9H-purin-6-yl)]pyrrolidin-3-yl}methyl(methyl)amino}piperidin-1-yl}methyl}-1H-benzo[d]imidazol-5-yl}pivalamide;

[0085] N-{2-{{4-{{[3-hydroxy-1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)]pyrrolidin-3-yl}methyl(methyl)amino}piperidin-1-yl}methyl}-1H-benzo[d]imidazol-5-yl}pivalamide;

[0086] 2-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide;

[0087] N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}acetamide;

[0088] N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}acetamide;

[0089] 2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide;

[0090] 3-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide;

[0091] N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}propanamide;

[0092] N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}propanamide;

[0093] 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide;

[0094] 3-{3-{2-{2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}acetamide}ethyl}ureido}thiophene-2-carboxylic acid methyl ester;

[0095] 3-{3-{2-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}propionamide}ethyl}ureido}thiophene-2-carboxylic acid methyl ester;

[0096] 2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(pyridin-3-yl)ureido]ethyl}acetamide;

[0097] 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(pyridin-3-yl)ureido]ethyl}propanamide;

[0098] 2-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide;

[0099] N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}acetamide;

[0100] N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}acetamide;

[0101] 2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide;

[0102] 3-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide;

[0103] N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanamide;

[0104] N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanamide;

[0105] 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide;

[0106] N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(2-chloro-9-methyl-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanamide;

[0107] 3-{{[1-(2-amino-9-methyl-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide.

[0108] The 1,3-disubstituted tetrahydropyrrole derivative forms a pharmaceutically acceptable salt with an acid, and the acid is: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid or benzoic acid.

[0109] The present invention provides a method for preparing the 1,3-disubstituted tetrahydropyrrole derivative or a pharmaceutically acceptable salt thereof, wherein the method is selected from one of method A, method B and method C;

[0110] Method A:

[0111]

[0112] Step 1: Compound 1-tert-butyloxycarbonyl-3-pyrrolidone undergoes a nucleophilic addition reaction with nitromethane under alkaline conditions to generate intermediate M1;

[0113] Step 2: Intermediate M1 undergoes reduction reaction in the presence of a reducing agent to generate intermediate M2;

[0114] Step 3: Different substituted aryl or heteroaryl primary amines react with ethylenediamine in the presence of carbonyldiimidazole (CDI) to generate intermediate M4;

[0115] Step 4: In the presence of a condensing agent, the intermediate M4 undergoes a condensation reaction with bromoacetic acid to generate an intermediate M5; or the intermediate M4 undergoes a condensation reaction with acrylic acid to generate an intermediate M6;

[0116] Step 5: Intermediate M2 and intermediate M3 undergo a reductive amination reaction in the presence of a reducing agent to generate intermediate M7; or intermediate M5 and intermediate M2 undergo an N-alkylation reaction under alkaline conditions to generate intermediate M7; or intermediate M5 and intermediate M2 undergo a Michael addition reaction to generate intermediate M7;

[0117] Step 6: Intermediate M7 is subjected to reductive amination reaction, Michael addition reaction or N-alkylation reaction in the presence of a base to generate intermediate M8;

[0118] Step 7: The intermediate M8 is deprotected from tert-butyloxycarbonyl (Boc) under acidic conditions to generate intermediate M9;

[0119] Step 8: Intermediate M9 undergoes N-alkylation reaction with differently substituted heteroaryl halides under alkaline conditions to obtain compound I.

[0120] Method B:

[0121]

[0122] Step 1: The intermediate M1 is deprotected from tert-butyloxycarbonyl (Boc) under acidic conditions to generate intermediate M10;

[0123] Step 2: Intermediate M10 undergoes N-alkylation reaction with differently substituted heteroaryl halides under alkaline conditions to generate intermediate M11;

[0124] Step 3: Intermediate M11 undergoes reduction reaction in the presence of a reducing agent to generate intermediate M12;

[0125] Step 4: Intermediate M12 and intermediate M5 undergo N-alkylation reaction under alkaline conditions to obtain compound I; or intermediate M5 and intermediate M2 undergo Michael addition reaction to obtain compound I.

[0126] Method C:

[0127]

[0128] Step 1: Intermediate M13 and intermediate M2 undergo reductive amination reaction in the presence of a reducing agent to generate intermediate M14;

[0129] Step 2: Intermediate M14 undergoes reductive amination reaction with various aldehyde-containing compounds in the presence of a reducing agent to generate intermediate M15;

[0130] Step 3: The intermediate M15 is deprotected from tert-butyloxycarbonyl (Boc) under acidic conditions to generate intermediate M16;

[0131] Step 4: Intermediate M16 undergoes N-alkylation reaction with differently substituted heteroaryl halides under alkaline conditions to obtain compound II.

[0132] The present invention provides a pharmaceutical composition comprising the 1,3-disubstituted tetrahydropyrrole derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0133] The present invention provides the use of the 1,3-disubstituted tetrahydropyrrole derivative or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition in preparing a drug for treating tumors.

[0134] Beneficial effects of the present invention

[0135] The present invention provides a DOT1L inhibitor with 1,3-disubstituted tetrahydropyrrole as the mother core, which fills the research gap; the 1,3-disubstituted tetrahydropyrrole derivative not only shows good inhibitory effect on both wild-type and mutant DOT1L proteins, but also has better in vitro efficacy than the existing DOT1L WT Compared with most existing DOT1L inhibitors WTInhibitors, the 1,3-disubstituted tetrahydropyrrole derivatives provided by the present invention avoid metabolically unstable adenosine fragments in their structures, so that they are not easily recognized and rapidly metabolized by multiple enzymes in the body, improve pharmacokinetic properties, and solve a major problem in the development of DOT1L inhibitors. In addition, the 1,3-disubstituted tetrahydropyrrole derivatives provided by the present invention have novel structures, simple preparation methods, and stable and easy-to-store products, providing highly potential candidate molecules for DOT1L drug discovery. DETAILED DESCRIPTION

[0136] The present invention is further described below in conjunction with specific implementation cases, but the present invention is not limited to these embodiments.

[0137] In the following examples, the H NMR spectra of the compounds were measured using Bruker ARX-400 / 600, and the mass spectra were measured using Agilent 1100 LC / MSD; all reagents used were of analytical or chemical purity.

[0138] Embodiment 1-17:

[0139]

[0140]

[0141] Embodiment 18-20:

[0142]

[0143]

[0144] Examples 21-28 and Examples 33-42:

[0145]

[0146]

[0147] Embodiment 29-32:

[0148]

[0149]

[0150] Example 1: 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(isopropyl)amino}propyl}urea (1)

[0151] The route looks like this:

[0152]

[0153] Step A1: Synthesis of Intermediate A-I

[0154] At room temperature, carbonyldiimidazole (7.06 g, 43.55 mmol) was dissolved in tetrahydrofuran (75 mL), 4-tert-butylaniline (5 g, 33.5 mmol) was added and stirred for 6 h. 3-aminopropanol (3.02 g, 40.2 mmol) was then added and the reaction was continued for 1.5 h. The reaction solution was evaporated to dryness, the residue was dissolved in ethyl acetate (300 mL), and the organic phase was then washed with 1 M hydrochloric acid aqueous solution (100 mL × 3), saturated sodium bicarbonate aqueous solution (100 mL) and saturated brine (100 mL), respectively. It was then dried over anhydrous sodium sulfate, filtered, and the organic phase was evaporated to dryness to obtain 8 g of a yellow oil with a yield of 95.3%. Analytical data: ESI-MS [M+H] + (m / z):251.17.

[0155] Step A2: Synthesis of intermediate A-II

[0156] Dissolve the intermediate A-Ⅰ (2g, 8mmol) in dry dichloromethane (80mL), cool to -10℃ under nitrogen protection, slowly add Dess-Martin oxidant (5.09g, 12mmol) in batches, and heat to 0℃ for 1.5h after addition. The reaction solution is quenched with saturated sodium thiosulfate solution (20mL) and saturated sodium bicarbonate solution (20mL), stirred at room temperature for 15min and filtered, and the filtrate is separated. The separated aqueous phase is extracted with dichloromethane (20mL×3), and the combined organic phases are dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a crude product. The crude product is recrystallized from ethyl acetate to obtain 0.9g of a white solid with a yield of 45%. Analytical data: ESI-MS[M+H] + (m / z):249.15.

[0157] Step A3: Synthesis of intermediate A-III

[0158] Dissolve 1-tert-butyloxycarbonyl-3-pyrrolidone (5 g, 27 mmol) in methanol (100 mL), add nitromethane (4.95 g, 81 mmol) and triethylamine (6.83 g, 67.5 mmol), and react at room temperature for 16 h. Evaporate the reaction solution to dryness to obtain a crude product, which is purified by column chromatography to obtain 6 g of a white solid with a yield of 90%. Analytical data: ESI-MS [M+Na] + (m / z):269.10.

[0159] Step A4: Synthesis of Intermediate A-IV

[0160] Dissolve intermediate A-Ⅲ (6 g, 8 mmol) in methanol (120 mL), add wet palladium carbon (0.3 g, 5%), and react at room temperature for 16 h under hydrogen conditions. Filter the reaction solution, wash the filter cake with methanol (10 mL), and evaporate the filtrate to dryness to obtain 4.96 g of colorless oil, with a yield of 94%. Analytical data: ESI-MS [M+H] + (m / z):217.10.

[0161] Step A5: Synthesis of Intermediate A-V

[0162] Add intermediate A-Ⅳ (1.5g, 6.94mmol) and intermediate A-Ⅱ (1.15g, 4.62mmol) to methanol (12mL), add 1 drop of glacial acetic acid and react at room temperature for 1h, then add sodium cyanoborohydride (0.5g, 7.96mmol) and continue to react for 6h. The reaction solution is quenched with saturated sodium bicarbonate aqueous solution (40mL) and extracted with dichloromethane (20mL×3). After combining the organic phases, wash with saturated brine (30mL), dry over anhydrous sodium sulfate, evaporate to dryness to obtain a crude product, which is then purified by column chromatography to obtain 1.2g of a white solid with a yield of 50.2%. Analytical data: ESI-MS [M+H] + (m / z):449.30.

[0163] Step A6: Synthesis of Intermediate A-VI

[0164] Dissolve intermediate A-V (0.45 g, 1.0 mmol) in acetonitrile (5 mL), add 2-iodopropane (0.85 g, 5.0 mmol) and potassium carbonate (0.42 g, 3.0 mmol), and react at 85°C for 48 h. Filter the reaction solution and evaporate to dryness to obtain a crude product, which is purified by column chromatography to obtain 0.25 g of a white solid with a yield of 50%. Analytical data: ESI-MS [M+H] + (m / z):491.10.

[0165] Step A7: Synthesis of Intermediate A-VII

[0166] Intermediate A-VI (0.3 g, 1.0 mmol) was dissolved in 4N HCl / ethyl acetate (6 mL) solution and stirred at room temperature for 1 h. After the reaction was completed, the reaction solution was evaporated to dryness to obtain 0.32 g of a white solid with a yield of 100%. Analytical data: ESI-MS [M+H] + (m / z):391.30.

[0167] Step A8: Synthesis of Example 1

[0168] Dissolve intermediate A-VII (70 mg, 0.18 mmol) and N,N-diisopropylethylamine (117 mg, 0.9 mmol) in isopropanol (2 mL), add 2,6-dichloropurine (102 mg, 0.54 mmol), and stir at 75 ° C for 48 h. Evaporate the reaction solution to dryness, dilute the residue with dichloromethane (5 mL), wash with saturated aqueous ammonium chloride solution (2 mL × 3), and dry over anhydrous sodium sulfate. Evaporate the organic phase to obtain a crude product, which is purified by column chromatography to obtain 56 mg of a white solid with a yield of 57.3%. Analytical data: mp: 203.6-207.3 ° C; ESI-MS [M+H] + (m / z):543.30; 1 H NMR (400 MHz, DMSO-d 6 )δ13.05(s,1H),8.26(s,1H),8.08(s,1H),7.26(d,J=8.7Hz,2H),7.20(d,J=8.7Hz, 2H),6.05(d,J=3.1Hz,1H),4.74(s,1H),4.36-4.10(m,1H),4.09-3.86(m,1H),3.79- 3.48(m,2H),3.11(q,J=6.5Hz,2H),3.01(q,J=6.6,5.8Hz,1H),2.56(s,2H),2.48(s ,1H),2.07-1.80(m,2H),1.55(q,J=7.1Hz,2H),1.23(s,10H),0.95(t,J=6.8Hz,6H).

[0169] Example 2: 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(isopropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea (2)

[0170] According to the method of step A8 in Example 1, intermediate A-VII was used as a raw material to undergo N-alkylation reaction with 3-bromo-8-chloroimidazo[1,2-a]pyrazine to obtain Example 2 with a yield of 47.3%. Analytical data: mp: 207.4-210.9°C; ESI-MS [M+H] + (m / z):586.30; 1 H NMR (400 MHz, DMSO-d 6)δ8.26(s,1H),7.67(s,1H),7.51(d,J=4.6Hz,1H),7.42(d,J=4.6Hz,1H),7.26(d,J =8.8Hz,2H),7.20(d,J=8.8Hz,2H),6.06(t,J=5.6Hz,1H),4.68(s,1H),3.90(d,J=12 7.3Hz,4H),3.11(q,J=6.5Hz,2H),3.06-2.94(m,1H),2.56(s,3H),1.99(dd,J=12.2, 8.2Hz, 1H), 1.87 (s, 1H), 1.56 (p, J = 6.8Hz, 2H), 1.23 (s, 10H), 0.96 (t, J = 6.5Hz, 6H).

[0171] Example 3: 1-[4-(tert-butyl)phenyl]-3-{3-{{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}(3-morpholinopropyl)amino}propyl}urea (3)

[0172] The route looks like this:

[0173]

[0174] Step A9: Synthesis of Intermediate A-VIII

[0175] Dissolve the intermediate A-V (0.6 g, 1.34 mmol) and 3-morpholinopropanal (0.38 g, 2.66 mmol) in methanol (9 mL), add 1 drop of glacial acetic acid and react at room temperature for 1 h, then add sodium cyanoborohydride (0.17 g, 2.66 mmol) and continue to react for 6 h. The reaction solution was quenched with saturated sodium bicarbonate aqueous solution (40 mL) and extracted with dichloromethane (20 mL × 3). The organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and evaporated to dryness to obtain a crude product, which was then purified by column chromatography to obtain 0.24 g of a white solid with a yield of 31%. Analytical data: ESI-MS [M+H] + (m / z):576.50.

[0176] Step A10: Synthesis of Intermediate A-IX

[0177] According to the method of step A7 in Example 1, intermediate A-VIII was used as the raw material and intermediate A-IX was prepared by deprotection reaction with a yield of 100%. Analytical data: ESI-MS [M+H] + (m / z):476.40.

[0178] Step A11: Synthesis of Example 3

[0179] According to the method of step A8 in Example 1, intermediate A-IX was used as a raw material to undergo N-alkylation reaction with 6-chloropurine to obtain Example 3 with a yield of 47%. Analytical data: mp: 214.5-217.9°C; ESI-MS [M+H] + (m / z):594.40; 1 HNMR (400 MHz, DMSO-d 6 )δ12.89(s,1H),8.29(s,1H),8.16(s,1H),8.04(s,1H),7.27(d,J=8.8Hz ,2H),7.21(d,J=8.8Hz,2H),6.09(t,J=5.6Hz,1H),5.32(s,1H),4.60-3.6 1(m,4H),3.49(s,5H),3.11(q,J=6.5Hz,2H),2.59(d,J=8.2Hz,4H),2.32( d, J=9.2Hz, 6H), 1.93 (d, J=10.2Hz, 2H), 1.68-1.49 (m, 4H), 1.23 (s, 10H).

[0180] Example 4: 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholinopropyl)amino}propyl}urea (4)

[0181] According to the method of step A11 in Example 3, intermediate A-IX was used as a raw material to undergo N-alkylation reaction with 2,6-dichloropurine to obtain Example 4 with a yield of 58.6%. Analytical data: mp: 215.8-218.7°C; ESI-MS [M+H] + (m / z): 628.40; 1 HNMR (400 MHz, DMSO-d 6)δ13.07(s,1H),8.28(s,1H),8.08(d,J=5.6Hz,1H),7.27(d,J=9.1Hz,2H),7.21(d,J=8.8H z,2H),6.07(q,J=5.0Hz,1H),5.33(s,1H),4.37-4.13(m,1H),3.98(dd,J=58.8,10.5Hz,1H) ,3.68(dd,J=22.7,11.7Hz,2H),3.49(dd,J=11.1,4.5Hz,5H),3.11(q,J=6.5Hz,2H),2.59( d,J=3.7Hz,4H),2.46-2.19(m,6H),2.05-1.84(m,2H),1.58(t,J=6.6Hz,4H),1.23(s,10H).

[0182] Example 5: 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholinopropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea (5)

[0183] According to the method of step A11 in Example 3, intermediate A-IX was used as a raw material to undergo N-alkylation reaction with 3-bromo-8-chloroimidazo[1,2-a]pyrazine to obtain Example 5 with a yield of 43.1%. Analytical data: mp: 217.9-221.3°C; ESI-MS [M+H] + (m / z): 671.30; 1 H NMR (400 MHz, DMSO-d 6 )δ8.26(s,1H),7.66(s,1H),7.52(d,J=4.6Hz,1H),7.42(d,J=4.6Hz,1H),7.27 (d,J=8.8Hz,2H),7.20(d,J=8.8Hz,2H),6.06(t,J=5.6Hz,1H),5.21(s,1H),3. 76(s,4H),3.48(d,J=4.8Hz,5H),3.11(q,J=6.5Hz,2H),2.57(d,J=3.3Hz,4H), 2.29(d,J=6.9Hz,6H),1.99-1.84(m,2H),1.56(h,J=6.9Hz,4H),1.23(s,10H).

[0184] Example 6: 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholinyl-3-oxopropyl)amino}propyl}urea (6)

[0185] The route looks like this:

[0186]

[0187] Step A12: Synthesis of Intermediate A-X

[0188] Dissolve intermediate A-V (0.33 g, 0.73 mmol) and 4-acryloylmorpholine (0.13 g, 0.88 mmol) in acetonitrile (9 mL) and react at 50°C for 24 h. Evaporate the reaction solution to dryness to obtain a crude product, which is then purified by column chromatography to obtain 0.3 g of a white solid with a yield of 69.8%. Analytical data: ESI-MS [M+H] + (m / z):590.40.

[0189] Step A13: Synthesis of Intermediate A-XI

[0190] According to the method of step A7 in Example 1, intermediate A-X was used as the raw material and intermediate A-XI was obtained by deprotection reaction with a yield of 100%. Analytical data: ESI-MS [M+H] + (m / z):490.40.

[0191] Step A14: Synthesis of Example 6

[0192] According to the method of step A8 in Example 1, intermediate A-XI was used as a raw material to undergo N-alkylation reaction with 2,6-dichloropurine to obtain Example 6 with a yield of 64.2%. Analytical data: mp: 226.1-229.4°C; ESI-MS [M+H] + (m / z): 642.30; 1 HNMR (400 MHz, DMSO-d 6 )δ13.06(s,1H),8.27(s,1H),8.08(d,J=7.8Hz,1H),7.32-7.24(m,2H),7.21(d,J=8.8H z,2H),6.09(q,J=5.3Hz,1H),5.01(d,J=18.2Hz,1H),4.39-3.59(m,4H),3.51(t,J=4.9H z,4H),3.43(d,J=11.3Hz,4H),3.11(q,J=6.5Hz,2H),2.83(d,J=8.3Hz,2H),2.58(d,J= 27.7Hz, 4H), 2.49-2.43 (m, 1H), 2.05-1.82 (m, 2H), 1.57 (d, J = 8.8Hz, 2H), 1.23 (s, 10H).

[0193] Example 7: 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholinyl-3-oxopropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea (7)

[0194] According to the method of step A14 in Example 6, intermediate A-XI was used as a raw material to undergo N-alkylation reaction with 3-bromo-8-chloroimidazo[1,2-a]pyrazine to obtain Example 7 with a yield of 49.7%. Analytical data: mp: 222.6-225.3°C; ESI-MS [M+H] + (m / z): 685.20; 1 H NMR (400 MHz, DMSO-d 6 )δ8.26(s,1H),7.66(s,1H),7.52(d,J=4.6Hz,1H),7.42(d,J=4.5Hz,1H),7.27(d,J=8.8Hz,2 H),7.20(d,J=8.8Hz,2H),6.08(t,J=5.5Hz,1H),4.94(s,1H),3.70(s,4H),3.54-3.48(m,4H), 3.42(d,J=4.9Hz,4H),3.11(q,J=6.5Hz,2H),2.82(q,J=8.0,6.9Hz,2H),2.57(d,J=23.8Hz,4 H), 2.48 (d, J = 6.9 Hz, 1H), 1.94 (dd, J = 25.4, 15.5 Hz, 2H), 1.56 (p, J = 6.8 Hz, 2H), 1.23 (s, 10H).

[0195] Example 8: 1-[4-(tert-butyl)phenyl]-3-{3-{{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}(3-morpholinyl-3-oxopropyl)amino}propyl}urea (8)

[0196] According to the method of step A14 in Example 6, intermediate A-XI was used as a raw material to undergo N-alkylation reaction with 6-chloropurine to obtain Example 8 with a yield of 40.7%. Analytical data: mp: 214.2-217.6°C; ESI-MS [M+H] + (m / z): 608.40; 1 HNMR (400 MHz, DMSO-d 6)δ12.88(s,1H),8.27(s,1H),8.16(s,1H),8.04(s,1H),7.27(d,J=8.7Hz,2H),7.21( d,J=8.8Hz,2H),6.09(t,J=5.5Hz,1H),4.96(d,J=12.5Hz,1H),4.40-3.60(m,4H),3.5 0(d,J=3.6Hz,4H),3.41(s,4H),3.11(q,J=6.4Hz,2H),2.82(d,J=6.2Hz,2H),2.58(d, J=24.4Hz, 4H), 2.46 (s, 1H), 1.96 (d, J= 25.5Hz, 2H), 1.63-1.50 (m, 2H), 1.23 (s, 10H).

[0197] Example 9: 1-[4-(tert-butyl)phenyl]-3-{3-{{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}urea (9)

[0198] The route looks like this:

[0199]

[0200] Step A15: Synthesis of Intermediate A-XII

[0201] Dissolve intermediate A-V (0.36 g, 0.8 mmol) and 4-acryloylthiomorpholine (0.15 g, 0.96 mmol) in acetonitrile (8 mL) and react at 50°C for 24 h. Evaporate the reaction solution to dryness to obtain a crude product, which is then purified by column chromatography to obtain 0.33 g of a white solid with a yield of 68.1%. Analytical data: ESI-MS [M+H] + (m / z):606.30.

[0202] Step A16: Synthesis of Intermediate A-XIII

[0203] According to the method of step A7 in Example 1, intermediate A-XIII was prepared by deprotection reaction with intermediate A-XII as raw material, with a yield of 100%. Analytical data: ESI-MS [M+H] + (m / z):506.40.

[0204] Step A17: Synthesis of Example 9

[0205] According to the method of step A8 in Example 1, intermediate A-XIII was used as a raw material to undergo N-alkylation reaction with 6-chloropurine to obtain Example 9 with a yield of 54.6%. Analytical data: mp: 218.3-221.3°C; ESI-MS [M+H]+ (m / z): 624.40; 1 HNMR (400 MHz, DMSO-d 6 )δ12.88(s,1H),8.27(s,1H),8.16(s,1H),8.04(s,1H),7.27(d,J=8.7Hz,2 H),7.21(d,J=8.8Hz,2H),6.09(t,J=5.6Hz,1H),4.97(s,1H),4.39-3.88(m ,2H),3.67(s,6H),3.11(q,J=6.5Hz,2H),2.94-2.73(m,2H),2.61(s,2H),2 .59-2.52(m,4H),1.97(d,J=33.9Hz,2H),1.66-1.48(m,2H),1.23(s,10H).

[0206] Example 10: 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}urea (10)

[0207] According to the method of step A17 in Example 9, intermediate A-XIII was used as a raw material to undergo N-alkylation reaction with 2,6-dichloropurine to obtain Example 10 with a yield of 72%. Analytical data: mp: 219.5-223.0°C; ESI-MS [M+H] + (m / z): 658.30; 1 H NMR (400 MHz, DMSO-d 6 )δ13.06(s,1H),8.27(d,J=3.3Hz,1H),8.07(d,J=6.7Hz,1H),7.27(dd,J=8.8,2.5Hz,2H ),7.23-7.17(m,2H),6.08(q,J=5.3Hz,1H),5.00(d,J=18.0Hz,1H),4.38-3.89(m,2H),3. 65(dd,J=25.3,10.5Hz,6H),3.11(q,J=6.4Hz,2H),2.82(s,2H),2.61(s,2H),2.54(s,4H) ,2.47(d,J=10.2Hz,3H),1.93(ddd,J=33.1,21.6,7.6Hz,2H),1.56(s,2H),1.23(s,10H).

[0208] Example 11: 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}urea (11)

[0209] According to the method of step A17 in Example 9, intermediate A-XIII was used as a raw material to undergo N-alkylation reaction with 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine to obtain Example 11 with a yield of 63%. Analytical data: mp: 218.3-221.7°C; ESI-MS [M+H] + (m / z): 658.30; 1 HNMR (400 MHz, DMSO-d 6 )δ13.57(s,1H),8.27(d,J=4.3Hz,1H),8.15(d,J=29.8Hz,1H),7.27(ddt,J=7.0,4.6,2.3Hz,2H),7.2 0(dd,J=8.8,2.1Hz,2H),6.09(t,J=5.6Hz,1H),5.08(d,J=22.8Hz,1H),3.85(ddd,J=37.6,8.1,4.5Hz ,2H),3.76-3.48(m,6H),3.12(p,J=6.4Hz,2H),2.83(tp,J=13.3,7.2Hz,2H),2.63(d,J=10.9Hz,2H), 2.55(dt,J=9.8,4.6Hz,4H),2.48(s,3H),2.16-1.84(m,2H),1.57(p,J=7.9,7.3Hz,2H),1.23(s,10H).

[0210] Example 12: 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea (12)

[0211] According to the method of step A17 in Example 9, intermediate A-XIII was used as a raw material to undergo N-alkylation reaction with 3-bromo-8-chloroimidazo[1,2-a]pyrazine to obtain Example 12 with a yield of 59.1%. Analytical data: mp: 220.9-224.8°C; ESI-MS [M+H] + (m / z):701.30; 1 H NMR (400 MHz, DMSO-d 6)δ8.26(s,1H),7.66(s,1H),7.52(d,J=4.5Hz,1H),7.42(d,J=4.6Hz,1H),7.31-7 .24(m,2H),7.23-7.17(m,2H),6.08(t,J=5.6Hz,1H),4.94(s,1H),3.68(q,J=5.1H z,8H),3.11(q,J=6.5Hz,2H),2.81(t,J=6.0Hz,2H),2.60(s,2H),2.58-2.52(m,4 H), 2.48 (d, J = 8.1Hz, 3H), 2.03-1.82 (m, 2H), 1.56 (p, J = 6.9Hz, 2H), 1.23 (s, 10H).

[0212] Example 13: Methyl 3-{{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}amino}propanoate (13)

[0213] The route looks like this:

[0214]

[0215] Step A18: Synthesis of Intermediate A-XIV

[0216] Dissolve intermediate A-V (0.45 g, 1 mmol) and methyl acrylate (0.1 g, 1.2 mmol) in acetonitrile (9 mL) and react at 50°C for 24 h. Evaporate the reaction solution to dryness to obtain a crude product, which is then purified by column chromatography to obtain 0.3 g of a white solid with a yield of 56%. Analytical data: ESI-MS [M+H] + (m / z):535.34.

[0217] Step A19: Synthesis of Intermediate A-XV

[0218] According to the method of step A7 in Example 1, intermediate A-XIV was used as a raw material and intermediate A-XV was prepared by deprotection reaction with a yield of 100%. Analytical data: ESI-MS [M+H] + (m / z):435.40.

[0219] Step A20: Synthesis of Example 13

[0220] According to the method of step A8 in Example 1, intermediate A-XV was used as a raw material to undergo N-alkylation reaction with 6-chloropurine to obtain Example 13 with a yield of 32.3%. Analytical data: mp: 204.5-207.2°C; ESI-MS [M+H] + (m / z):553.30;1 HNMR (400 MHz, DMSO-d 6 )δ12.88(s,1H),8.27(s,1H),8.16(s,1H),8.05(s,1H),7.27(d,J=8.7Hz,2H),7 .21(d,J=8.8Hz,2H),6.06(t,J=5.6Hz,1H),4.71(s,1H),4.42-3.60(m,4H),3.58 (s,4H),3.09(q,J=6.5Hz,2H),2.85(t,J=7.2Hz,2H),2.61(s,2H),2.56(t,J=7.3 Hz, 2H), 2.47 (s, 1H), 1.89 (d, J = 30.0Hz, 2H), 1.56 (p, J = 6.7Hz, 2H), 1.23 (s, 9H).

[0221] Example 14: 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}amino}propionic acid methyl ester (14)

[0222] According to the method of step A20 in Example 13, intermediate A-XV was used as a raw material to undergo N-alkylation reaction with 3-bromo-8-chloroimidazo[1,2-a]pyrazine to obtain Example 14 with a yield of 33.1%. Analytical data: mp: 203.1-205.9°C; ESI-MS [M+H] + (m / z): 630.30; 1 H NMR (400 MHz, DMSO-d 6 )δ8.26(s,1H),7.67(s,1H),7.52(d,J=4.5Hz,1H),7.42(d,J=4.6Hz,1H),7.3 4-7.23(m,2H),7.23-7.12(m,2H),6.06(t,J=5.7Hz,1H),4.70(s,1H),3.58(s ,8H),3.09(q,J=6.5Hz,2H),2.85(t,J=7.1Hz,2H),2.60(s,2H),2.56(t,J=7. 2Hz, 2H), 2.47 (s, 1H), 2.02-1.81 (m, 2H), 1.56 (p, J = 6.9Hz, 2H), 1.23 (s, 9H).

[0223] Example 15: Methyl 3-{{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanoate (15)

[0224] According to the method of step A20 in Example 13, intermediate A-XV was used as a raw material to undergo N-alkylation reaction with 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine to obtain Example 15 with a yield of 55.6%. Analytical data: mp: 205.3-208.5°C; ESI-MS [M+H] + (m / z):587.30; 1 H NMR (400 MHz, DMSO-d 6 )δ13.58(d,J=8.1Hz,1H),8.27(d,J=5.7Hz,1H),8.13(d,J=21.2Hz,1H),7.33-7.16(m,4H), 6.06(td,J=5.7,2.9Hz,1H),4.85(d,J=31.9Hz,1H),3.85(ddd,J=37.5,8.2,4.6Hz,2H),3.75 -3.36(m,6H),3.10(qd,J=6.5,2.7Hz,2H),2.94-2.77(m,2H),2.70-2.61(m,2H),2.56(h,J=6 .7, 4.7Hz, 2H), 2.48 (d, J = 7.0Hz, 1H), 2.13-1.86 (m, 2H), 1.57 (p, J = 6.7Hz, 2H), 1.23 (s, 9H).

[0225] Example 16: Methyl 3-{{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanoate (16)

[0226] According to the method of step A20 in Example 13, intermediate A-XV was used as a raw material to undergo N-alkylation reaction with 2,6-dichloropurine to obtain Example 16 with a yield of 41.5%. Analytical data: mp: 206.1-209.7°C; ESI-MS [M+H] + (m / z):587.30; 1 H NMR (400 MHz, DMSO-d 6)δ13.07(s,1H),8.28(d,J=3.1Hz,1H),8.08(d,J=3.3Hz,1H),7.43-7.14(m,4 H),6.07(t,J=3.6Hz,1H),4.77(d,J=15.0Hz,1H),4.38-3.82(m,2H),3.80-3.3 8(m,6H),3.09(q,J=6.5Hz,2H),2.96-2.76(m,2H),2.61(s,2H),2.55(t,J=5.7 Hz,2H),2.49-2.43(m,1H),2.10-1.81(m,2H),1.67-1.49(m,2H),1.23(s,9H).

[0227] Example 17: Methyl 3-{{[3-acetoxy-1-(2-chloro-9H-purin-6-yl)pyrrolidin-3-yl]methyl}{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}amino}propanoate (17)

[0228] Step A21: Synthesis of Example 17

[0229] Example 16 (100 mg, 0.17 mmol) and triethylamine (35 mg, 0.34 mmol) were dissolved in dichloromethane (2 mL), and acetyl chloride (27 mg, 0.34 mmol) was added dropwise under ice bath conditions. After the addition was completed, the mixture was stirred at room temperature for 3 h. The reaction solution was evaporated to dryness to obtain a crude product, which was purified by column chromatography to obtain 67 mg of a white solid with a yield of 63%. Analytical data: mp: 211.2-213.6°C; ESI-MS [M+H] + (m / z):629.30.

[0230] Example 18: N-{2-{{4-{{[3-hydroxy-1-(9H-purin-6-yl)]pyrrolidin-3-yl}methyl(methyl)amino}piperidin-1-yl}methyl}-1H-benzo[d]imidazol-5-yl}pivalamide (18)

[0231] The synthetic route is as follows:

[0232]

[0233] Step B1: Synthesis of intermediate B-Ⅰ

[0234] At room temperature, add 4-nitro-o-phenylenediamine (10.0 g, 0.07 mol), chloroacetic acid (12.3 g, 0.13 mol) and water (50 mL) to concentrated hydrochloric acid (133.0 mL), then heat to reflux and stir to react for 4 hours. The reaction solution was cooled to room temperature, adjusted to pH = 9 with ammonia water, and filtered to obtain an orange-red solid. The crude product was recrystallized with methanol to obtain 8.9 g of a light yellow solid with a yield of 64.5%. Analytical data: ESI-MS [M+H] + (m / z):212.09.

[0235] Step B2: Synthesis of intermediate B-II

[0236] At room temperature, the intermediate B-Ⅰ (9.0 g, 0.04 mol) was dissolved in tetrahydrofuran (100.0 mL), 4-piperidone hydrochloride hydrate (7.8 g, 0.08 mol) and anhydrous potassium carbonate (10.8 g, 0.08 mol) were added, and the mixture was reacted at 50°C for 6 h. Most of the tetrahydrofuran in the reaction solution was evaporated, and the residue was poured into water (100.0 mL), stirred for 30 min, and then filtered to obtain 8.0 g of a light yellow solid with a yield of 74.9%. Analytical data: ESI-MS [M+H] + (m / z):275.39.

[0237] Step B3: Synthesis of intermediate B-III

[0238] At room temperature, add intermediate B-Ⅱ (3.0g, 0.011mol) and reduced iron powder (3.1g, 0.055mol) to methanol (30.0mL), heat to 65℃ and react for 30min, then add ammonium chloride (3.0g, 0.057mol) in water (15.0mL), continue stirring and reacting for 1.5h. Filter while hot, wash with hot methanol, and evaporate the filtrate. Pour the residue into water (30mL) and stir, adjust the pH to 9 with saturated sodium bicarbonate solution, extract the aqueous phase with a mixed solvent (DCM: MeOH = 8: 1, 30.0mL × 3), combine the organic phases and evaporate to dryness to obtain 1.8g of yellow solid, with a yield of 65.2%. Analytical data: ESI-MS [M+H] + (m / z):245.12.

[0239] Step B4: Synthesis of intermediate B-IV

[0240] At room temperature, add intermediate B-Ⅲ (3.8g, 0.004mol) and triethylamine (4.7g, 0.012mol) to isopropanol (38.0mL), and slowly add pivaloyl chloride (3.7g, 0.008mol) at 0℃. After the addition, raise the temperature to 35℃ and stir to react overnight. Evaporate the reaction solution, pour the residue into water (40.0mL), extract with ethyl acetate (30.0mL×3), combine the organic phases and evaporate to dryness to obtain 3.6g of yellow solid, with a yield of 72.0%. Analytical data: ESI-MS [M+H] + (m / z):329.19.

[0241] Step B5: Synthesis of Intermediate B-V

[0242] Intermediate A-IV (2g, 9.15mmol) and intermediate B-IV (2g, 6.1mmol) were added to a mixed solution of methanol and dichloromethane (10mL:5mL), 1 drop of glacial acetic acid was added and reacted at room temperature for 1h, followed by the addition of sodium cyanoborohydride (0.6g, 9.15mmol) and the reaction was continued for 6h. The reaction solution was quenched with saturated aqueous sodium bicarbonate solution (30mL) and extracted with dichloromethane (20mL×6). The organic phases were combined and washed with saturated brine (30mL), dried over anhydrous sodium sulfate, and evaporated to dryness to obtain a crude product, which was then purified by column chromatography to obtain 0.9g of a white solid with a yield of 56.3%. Analytical data: ESI-MS [M+H] + (m / z):529.30.

[0243] Step B6: Synthesis of Intermediate B-VI

[0244] Add intermediate B-V (0.5 g, 0.95 mmol) and 37% formaldehyde aqueous solution (0.15 g, 1.9 mmol) to methanol solution (5 mL), add 1 drop of glacial acetic acid, stir at room temperature for 1 h, then add sodium cyanoborohydride (0.12 g, 1.9 mmol), and react at room temperature for 12 h. The reaction solution was quenched with saturated sodium bicarbonate aqueous solution (20 mL) and extracted with dichloromethane (20 mL × 3). After combining the organic phases, wash with saturated brine (20 mL), dry over anhydrous sodium sulfate, and evaporate to dryness to obtain 0.52 g of a white solid with a yield of 100%. Analytical data: ESI-MS [M+H] + (m / z):543.40.

[0245] Step B7: Synthesis of Intermediate B-VII

[0246] Dissolve the intermediate B-VI (0.3 g, 0.55 mmol) in 4N HCl / ethyl acetate (6 mL) solution and stir at room temperature for 1 h. After the reaction is complete, evaporate the reaction solution to dryness to obtain 0.35 g of a white solid with a yield of 100%. Analytical data: ESI-MS [M+H] + (m / z):443.50.

[0247] Step B8: Synthesis of Example 18

[0248] Dissolve intermediate B-VII (0.1 g, 0.23 mmol) and N,N-diisopropylethylamine (0.15 g, 1.15 mmol) in isopropanol (2 mL), add 6-chloropurine (0.11 g, 0.69 mmol), and stir at 75 ° C for 10 h. Evaporate the reaction solution to dryness, dilute the residue with dichloromethane (5 mL), wash with saturated aqueous ammonium chloride solution (3 mL × 3), dry with anhydrous sodium sulfate, evaporate the organic phase to obtain a crude product, and purify the crude product by column chromatography to obtain 61 mg of a white solid with a yield of 47.6%. Analytical data: mp: 238.1-239.9 ° C; ESI-MS [M+H] + (m / z):561.50; 1 H NMR (600 MHz, DMSO-d 6 )δ12.90(s,1H),12.30-11.96(m,1H),9.13(s,1H),8.16(s,1H),8.05(s,1H),7 .88(s,1H),7.39(dd,J=19.3,10.1Hz,1H),7.27(d,J=14.9Hz,1H),5.09-3.98(m ,5H),3.65(s,3H),3.48(s,2H),2.89(d,J=11.1Hz,1H),2.61(s,2H),2.39(d,J= 27.4Hz, 3H), 1.98 (d, J = 63.8Hz, 3H), 1.81-1.41 (m, 4H), 1.24 (d, J = 11.5Hz, 9H).

[0249] Example 19: N-{2-{{4-{{[3-hydroxy-1-(2-chloro-9H-purin-6-yl)]pyrrolidin-3-yl}methyl(methyl)amino}piperidin-1-yl}methyl}-1H-benzo[d]imidazol-5-yl}pivalamide (19)

[0250] According to the method of step B8 in Example 18, intermediate B-VII was used as a raw material to undergo N-alkylation reaction with 2,6-dichloropurine to obtain Example 19 with a yield of 60.5%. Analytical data: mp: 236.8-238.4°C; ESI-MS [M+H] +(m / z):595.70; 1 H NMR (600 MHz, DMSO-d 6 )δ13.10(s,1H),12.14(s,1H),9.13(s,1H),7.98(d,J=122.5Hz,2H),7.33(d,J=55.9Hz,2H),5.18-3.83(m,4H),3.70(d,J=34.2Hz,3H ), 3.51 (t, J = 26.8Hz, 2H), 2.91 (s, 2H), 2.70 (s, 1H), 2.42 (s, 2H), 1.98 (dd, J = 96.0, 34.4Hz, 4H), 1.63 (d, J = 105.9Hz, 4H), 1.23 (s, 9H).

[0251] Example 20: N-{2-{{4-{{[3-hydroxy-1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)]pyrrolidin-3-yl}methyl(methyl)amino}piperidin-1-yl}methyl}-1H-benzo[d]imidazol-5-yl}pivalamide (20)

[0252] According to the method of step B8 in Example 18, intermediate B-VII was used as a raw material to undergo N-alkylation reaction with 3-bromo-8-chloroimidazo[1,2-a]pyrazine to obtain Example 20 with a yield of 50.9%. Analytical data: mp: 232.1-233.9°C; ESI-MS [M+H] + (m / z):638.30.

[0253] Example 21: 2-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide (21)

[0254] The route looks like this:

[0255]

[0256] Step C1: Synthesis of Intermediate C-Ⅰ

[0257] At room temperature, carbonyldiimidazole (7.06 g, 43.55 mmol) was dissolved in tetrahydrofuran (75 mL), 4-tert-butylaniline (5 g, 33.5 mmol) was added and stirred for 6 h. Ethylenediamine (6.04 g, 100.5 mmol) was then added and the reaction was continued for 1.5 h. The reaction solution was evaporated to dryness, the residue was dissolved in ethyl acetate (300 mL), and the organic phase was then washed with 1 M hydrochloric acid aqueous solution (100 mL × 3), saturated sodium bicarbonate aqueous solution (100 mL) and saturated brine (100 mL), respectively, and then dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness to obtain a crude product, which was purified by column chromatography to obtain 5.7 g of a white solid with a yield of 72.6%. Analytical data: ESI-MS [M+H] + (m / z):236.20.

[0258] Step C2: Synthesis of intermediate C-II

[0259] At room temperature, bromoacetic acid (2.03 g, 14.59 mmol), EDCI (3.28 g, 18.24 mmol) and HOBt (2.47 g, 18.24 mmol) were added to dichloromethane (45 mL), stirred at room temperature for 1 h, and then intermediate C-I (2.8 g, 12.16 mmol) was added, and the reaction was continued to stir for 16 h. The reaction solution was quenched with water (100 mL), and the aqueous phase was extracted with dichloromethane (50 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness to obtain a crude product. The crude product was purified by column chromatography to obtain 2.8 g of a colorless oil with a yield of 63.6%. Analytical data: ESI-MS [M+H] + (m / z):356.10.

[0260] Step C3: Synthesis of intermediate C-III

[0261] At room temperature, intermediate C-Ⅱ (2.8 g, 7.86 mmol), intermediate A-Ⅳ (3.40 g, 15.72 mmol) and triethylamine (1.59 g, 15.72 mmol) were added to methanol (50 mL), and then the temperature was raised to 60°C and stirred for 48 h. The reaction solution was evaporated to dryness to obtain a crude product, which was purified by column chromatography to obtain 2.1 g of a white solid with a yield of 54.7%. Analytical data: ESI-MS [M+H] + (m / z):492.30.

[0262] Step C4: Synthesis of intermediate C-IV

[0263] At room temperature, intermediate C-Ⅲ (2.1g, 4.30mmol) and 37% formaldehyde aqueous solution (0.7g, 8.60mmol) were added to methanol (30mL), and 1 drop of glacial acetic acid was added and stirred for 1h, followed by the addition of sodium cyanoborohydride (0.4g, 6.45mmol), and the reaction was continued with stirring for 3h. The reaction solution was quenched with saturated sodium bicarbonate aqueous solution (50mL), extracted with dichloromethane (30mL×3), and the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness to obtain a crude product, which was purified by column chromatography to obtain 1.9g of a white solid with a yield of 88.3%. Analytical data: ESI-MS [M+H] + (m / z):506.30.

[0264] Step C5: Synthesis of Intermediate C-V

[0265] Add intermediate C-IV (1.9 g, 3.76 mmol) to 4N HCl / EA solution (20 mL) and stir at room temperature for 1 h. Evaporate the reaction solution to dryness to obtain a crude product, which is purified by column chromatography to obtain 1.2 g of a white solid with a yield of 79.5%. Analytical data: ESI-MS [M+H] + (m / z):406.30.

[0266] Step C6: Synthesis of Example 21

[0267] Dissolve the intermediate C-V (0.3 g, 0.74 mmol) and N,N-diisopropylethylamine (0.28 g, 2.22 mmol) in isopropanol (3 mL), add 5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (0.26 g, 1.11 mmol), and stir at 60°C for 10 h. Evaporate the reaction solution to dryness, dilute the residue with dichloromethane (10 mL), wash with saturated aqueous ammonium chloride solution (3 mL × 3), dry with anhydrous sodium sulfate, evaporate the organic phase to obtain a crude product, and purify the crude product by column chromatography to obtain 67 mg of a white solid with a yield of 15.1%. Analytical data: mp: 247.3-249.0°C; ESI-MS [M+H] + (m / z):601.20.

[0268] Example 22: N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}acetamide (22)

[0269] According to the method of step C6 in Example 21, intermediate C-V was used as a raw material to undergo N-alkylation reaction with 2,6-dichloropurine to obtain Example 22 with a yield of 43.8%. Analytical data: mp: 244.7-246.3°C; ESI-MS [M+H]+ (m / z):558.30.

[0270] Example 23: N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}acetamide (23)

[0271] According to the method of step C6 in Example 21, intermediate C-V was used as a raw material to undergo N-alkylation reaction with 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine to obtain Example 23 with a yield of 23.5%. Analytical data: mp: 243.9-245.5°C; ESI-MS [M+H] + (m / z):558.30.

[0272] Example 24: 2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide (24)

[0273] According to the method of step C6 in Example 21, intermediate C-V was used as a raw material to undergo N-alkylation reaction with 3-bromo-8-chloroimidazo[1,2-a]pyrazine to obtain Example 24 with a yield of 52.7%. Analytical data: mp: 240.2-241.8°C; ESI-MS [M+H] + (m / z):601.20.

[0274] Example 25: 3-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide (25)

[0275] The route looks like this:

[0276]

[0277] Step C7: Synthesis of Intermediate C-VI

[0278] At room temperature, acrylic acid (1.05 g, 14.59 mmol), HATU (5.55 g, 14.59 mmol) and N, N-diisopropylethylamine (3.14 g, 24.32 mmol) were added to dichloromethane (50 mL), stirred at room temperature for 30 min, then intermediate C-I (2.8 g, 12.16 mmol) was added, and the reaction was continued for 6 h. The reaction solution was washed with water (20 mL), saturated ammonium chloride solution (20 mL), saturated sodium bicarbonate solution (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness to obtain a crude product, which was purified by column chromatography to obtain 1.9 g of a white solid with a yield of 53.3%. Analytical data: ESI-MS [M+H] + (m / z):290.20.

[0279] Step C8: Synthesis of Intermediate C-VII

[0280] At room temperature, intermediate C-VI (1.9 g, 6.57 mmol) and intermediate A-IV (2.84 g, 13.13 mmol) were added to methanol (50 mL), and then the temperature was raised to 60°C and stirred for 48 h. The reaction solution was evaporated to dryness to obtain a crude product, which was purified by column chromatography to obtain 2.3 g of a white solid with a yield of 69.4%. Analytical data: ESI-MS [M+H] + (m / z):506.30.

[0281] Step C9: Synthesis of Intermediate C-VIII

[0282] According to the method of step C4 in Example 21, intermediate C-VII was used as a raw material and subjected to a reductive amination reaction with 37% formaldehyde aqueous solution to obtain intermediate C-VIII with a yield of 46.2%. Analytical data: ESI-MS [M+H] + (m / z):520.30.

[0283] Step C10: Synthesis of Intermediate C-IX

[0284] According to the method of step C5 in Example 21, intermediate C-VIII was used as the raw material and deprotected to obtain intermediate C-IX with a yield of 100%. Analytical data: ESI-MS [M+H] + (m / z):420.30.

[0285] Step C11: Synthesis of Example 25

[0286] According to the method of step C6 in Example 21, intermediate C-IX was used as a raw material to undergo N-alkylation reaction with 5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine to obtain Example 25 with a yield of 11.8%. Analytical data: mp: 248.1-250.0°C; ESI-MS [M+H] + (m / z):615.20.

[0287] Example 26: N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}propanamide (26)

[0288] According to the method of step C11 in Example 25, intermediate C-IX was used as a raw material to undergo N-alkylation reaction with 2,6-dichloropurine to obtain Example 26 with a yield of 18.4%. Analytical data: mp: 245.5-247.3°C; ESI-MS [M+H] + (m / z):572.30.

[0289] Example 27: N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}propanamide (27)

[0290] According to the method of step C11 in Example 25, intermediate C-IX was used as a raw material to undergo N-alkylation reaction with 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine to obtain Example 27 with a yield of 13.6%. Analytical data: mp: 244.9-246.4°C; ESI-MS [M+H] + (m / z):572.30.

[0291] Example 28: 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide (28)

[0292] According to the method of step C11 in Example 25, intermediate C-IX was used as a raw material to undergo N-alkylation reaction with 3-bromo-8-chloroimidazo[1,2-a]pyrazine to obtain Example 28 with a yield of 17.9%. Analytical data: mp: 246.1-248.0°C; ESI-MS [M+H] + (m / z):615.20.

[0293] Example 29: 3-{3-{2-{2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}acetamide}ethyl}ureido}thiophene-2-carboxylic acid methyl ester (29)

[0294] According to the method of steps C1 to C6 in Example 21, methyl 3-amino-2-thiophenecarboxylate was used as the raw material, and Example 29 was prepared through 6 steps with a yield of 6.1%. Analytical data: mp: 253.2-254.9°C; ESI-MS [M+H] + (m / z):609.10.

[0295] Example 30: 3-{3-{2-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}propionamide}ethyl}ureido}thiophene-2-carboxylic acid methyl ester (30)

[0296] According to the method of steps C7 to C11 in Example 25, methyl 3-(3-(2-aminoethyl)ureido)thiophene-2-carboxylate was used as the raw material, and Example 30 was prepared through 5 steps with a yield of 6.5%. Analytical data: mp: 254.6-256.2°C; ESI-MS [M+H] + (m / z):623.10.

[0297] Example 31: 2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(pyridin-3-yl)ureido]ethyl}acetamide (31)

[0298] According to the method of steps C1 to C6 in Example 21, 3-amino-pyridine was used as the raw material and Example 29 was prepared through 6 steps with a yield of 8.5%. Analytical data: mp: 249.5-251.2°C; ESI-MS [M+H] + (m / z):546.10.

[0299] Example 32: 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(pyridin-3-yl)ureido]ethyl}propanamide (32)

[0300] According to the method of steps C7 to C11 in Example 25, 1-(2-aminoethyl)-3-(piperidin-3-yl)urea was used as the raw material and Example 32 was prepared through 5 steps with a yield of 11.3%. Analytical data: mp: 250.3-251.9°C; ESI-MS [M+H]+ (m / z):560.20.

[0301] Example 33: 2-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide (33)

[0302] The route looks like this:

[0303]

[0304] Step D1: Synthesis of intermediate D-I

[0305] Add intermediate A-III (5 g, 20.3 mmol) to 4N HCl / EA solution (50 mL) and stir at room temperature for 1 h. Evaporate the reaction solution to dryness to obtain 3.8 g of white solid with a yield of 100%. Analytical data: ESI-MS [M+H] + (m / z):147.10.

[0306] Step D2: Synthesis of intermediate D-II

[0307] Dissolve intermediate D-Ⅰ (0.9 g, 4.93 mmol) and N,N-diisopropylethylamine (1.91 g, 14.79 mmol) in isopropanol (10 mL), add 5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1.72 g, 7.39 mmol), and stir at 40°C for 10 h. Evaporate the reaction solution to dryness, dilute the residue with dichloromethane (10 mL), wash with saturated aqueous ammonium chloride solution (3 mL × 3), dry with anhydrous sodium sulfate, evaporate the organic phase to obtain a crude product, and purify the crude product by column chromatography to obtain 1.4 g of a white solid with a yield of 80.4%. Analytical data: ESI-MS [M+H] + (m / z):342.0.

[0308] Step D3: Synthesis of intermediate D-III

[0309] Dissolve the intermediate D-Ⅱ (1.4 g, 4.09 mmol) in methanol (30 mL), add wet palladium carbon (0.2 g, 5%), and react at room temperature for 16 h under hydrogen conditions. Filter the reaction solution, wash the filter cake with methanol (10 mL), and evaporate the filtrate to dryness to obtain 1.2 g of colorless oil, with a yield of 94%. Analytical data: ESI-MS [M+H] + (m / z):312.0.

[0310] Step D4: Synthesis of Example 33

[0311] At room temperature, intermediate C-Ⅱ (0.2g, 0.56mmol), intermediate D-Ⅲ (0.35g, 1.12mmol) and triethylamine (0.11g, 1.12mmol) were added to methanol (4mL), and then the temperature was raised to 60℃ and stirred for 48h. The reaction solution was evaporated to dryness to obtain a crude product, which was purified by column chromatography to obtain 82mg of a white solid with a yield of 25%. Analytical data: mp: 243.5-244.8℃; ESI-MS [M+H] + (m / z):587.20.

[0312] Example 34: N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}acetamide (34)

[0313] The route looks like this:

[0314]

[0315] Step D5: Synthesis of intermediate D-IV

[0316] According to the method of steps D2 to D3 in Example 33, intermediate D-I and 2,6-dichloropurine were used as raw materials to obtain intermediate D-IV through two-step reaction with a yield of 43.7%. Analytical data: ESI-MS [M+H] + (m / z):269.10.

[0317] Step D6: Synthesis of Example 34

[0318] According to the method of step D4 in Example 33, intermediate D-IV was used as a raw material and reacted with intermediate C-II to obtain Example 34 with a yield of 13.8%. Analytical data: mp: 242.1-244.1°C; ESI-MS [M+H] + (m / z):544.30.

[0319] Example 35: N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}acetamide (35)

[0320] The route looks like this:

[0321]

[0322] Step D7: Synthesis of Intermediate D-V

[0323] According to the method of steps D2 to D3 in Example 33, intermediate D-I and 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine were used as raw materials to obtain intermediate D-V in two steps with a yield of 48.2%. Analytical data: ESI-MS [M+H] + (m / z):269.10.

[0324] Step D8: Synthesis of Example 35

[0325] According to the method of step D4 in Example 33, intermediate D-V was used as a raw material and reacted with intermediate C-II through N-alkylation to obtain Example 35 with a yield of 10.5%. Analytical data: mp: 241.7-243.6°C; ESI-MS [M+H] + (m / z):544.30.

[0326] Example 36: 2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide (36)

[0327] The route looks like this:

[0328]

[0329] Step D9: Synthesis of intermediate D-VI

[0330] According to the method of steps D2 to D3 in Example 33, intermediate D-I and 3-bromo-8-chloroimidazo[1,2-a]pyrazine were used as raw materials to obtain intermediate D-VI in two steps with a yield of 51.9%. Analytical data: ESI-MS [M+H] + (m / z):312.0.

[0331] Step D10: Synthesis of Example 36

[0332] According to the method of step D4 in Example 33, intermediate D-VI was used as a raw material and reacted with intermediate C-II through N-alkylation to obtain Example 36 with a yield of 13.3%. Analytical data: mp: 244.2-245.8°C; ESI-MS [M+H] + (m / z):587.20.

[0333] Example 37: 3-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide (37)

[0334] Step D11: Synthesis of Example 37

[0335] At room temperature, intermediate C-VI (0.2 g, 0.69 mmol) and intermediate D-III (0.43 g, 1.38 mmol) were added to methanol (4 mL), and then the temperature was raised to 60°C and stirred for 48 h. The reaction solution was evaporated to dryness to obtain a crude product, which was purified by column chromatography to obtain 93 mg of a white solid with a yield of 22%. Analytical data: mp: 245.6-247.1°C; ESI-MS [M+H] + (m / z):601.20.

[0336] Example 38: N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanamide (38)

[0337] According to the method of step D11 in Example 37, intermediate D-IV was used as a raw material and reacted with intermediate C-VI to obtain Example 38 with a yield of 28.1%. Analytical data: mp: 246.9-248.1°C; ESI-MS [M+H] + (m / z):558.30.

[0338] Example 39: N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanamide (39)

[0339] According to the method of step D11 in Example 37, intermediate D-V was used as a raw material and reacted with intermediate C-VI to obtain Example 39 with a yield of 33.7%. Analytical data: mp: 246.0-247.7°C; ESI-MS [M+H] + (m / z):558.30.

[0340] Example 40: 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide (40)

[0341] According to the method of step D11 in Example 37, intermediate D-VI was used as a raw material and reacted with intermediate C-VI to obtain Example 40 with a yield of 26.3%. Analytical data: mp: 244.8-246.4°C; ESI-MS [M+H] + (m / z):601.20.

[0342] Example 41: N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(2-chloro-9-methyl-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanamide (41)

[0343] The route looks like this:

[0344]

[0345] Step D12: Synthesis of Intermediate D-VII

[0346] According to the method of steps D2 to D3 in Example 33, intermediate D-I and 2,6-dichloro-9-methyl-9H-purine were used as raw materials to obtain intermediate D-VII through two-step reaction with a yield of 63.7%. Analytical data: ESI-MS [M+H] + (m / z):283.10.

[0347] Step D13: Synthesis of Example 41

[0348] According to the method of step D11 in Example 37, intermediate D-VII was used as a raw material and reacted with intermediate C-VI to obtain Example 41 with a yield of 37.6%. Analytical data: mp: 239.6-241.3°C; ESI-MS [M+H] + (m / z):572.30.

[0349] Example 42: 3-{{[1-(2-amino-9-methyl-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide (42)

[0350] The route looks like this:

[0351]

[0352] Step D14: Synthesis of Intermediate D-VIII

[0353] According to the method of steps D2 to D3 in Example 33, intermediate D-I and 6-chloro-9-methyl-9H-purine-2-amine were used as raw materials to obtain intermediate D-VIII in two steps with a yield of 48.8%. Analytical data: ESI-MS [M+H] + (m / z):264.20.

[0354] Step D15: Synthesis of Example 42

[0355] According to the method of step D11 in Example 37, intermediate D-VIII was used as a raw material and reacted with intermediate C-VI to obtain Example 42 with a yield of 25.4%. Analytical data: mp: 243.2-245.1°C; ESI-MS [M+H] + (m / z):553.30.

[0356] Example 43: Antitumor activity test of the compounds prepared in the above examples

[0357] 1. In vitro anti-tumor cell activity

[0358] The compounds of Examples 1-42 of the present invention were tested for their ability to inhibit the in vitro human lung cancer cell line H460-DOT1L WT 、H460-DOT1L R231Q Effect of cell viability.

[0359] 1. Cell culture

[0360] RPMI 1640 medium was used as the basic culture medium, and the cell culture medium was prepared to contain 10% fetal bovine serum. 2 Culture in a 37°C incubator. Change the culture medium once a day. When the cells grow to 80%-90%, discard the original culture medium, rinse once with PBS, digest the cells with 0.25% trypsin, and subculture the cells according to experimental requirements.

[0361] 2. Drug preparation

[0362] The required volume of DMSO was calculated according to the molecular weight and the weighed mass of the compound to prepare a stock solution (100 mM) of the same concentration and stored at 4°C.

[0363] 3. Experimental steps

[0364] (1) Cell inoculation: Take cells in good growth state in the logarithmic growth phase, digest the cells with 0.25% trypsin, and then disperse them into a single cell suspension by pipetting with culture medium. After counting, inoculate them in a 96-well plate at an appropriate density (200 cells / well), 100 μL / well, and incubate at saturated humidity, 37°C, 5% CO 2 Culture in an incubator.

[0365] (2) Drug treatment: After 24 hours of cell culture, according to the experimental needs, 6 different concentration gradients of drugs were added, 10 μL per well, 3 replicates per group, and a blank group per plate. In this experiment, the test drugs were diluted to the corresponding concentration gradients of 100 μM, 10 μM, 1 μM, 0.1 μM, and 0.01 μM for addition, and the action time was 72 hours. After drug addition, the cells were incubated at 5% CO 2, culture in a 37°C constant temperature cell culture incubator for a corresponding period of time, and observe the cell status under an inverted microscope.

[0366] (3) Color development and colorimetry: When the drug reaches the action time point, add 10 μL of 2.5 mg / mL MTT solution to each well and continue incubation for 1-4 h. Terminate the incubation, discard the supernatant, add 100 μL of dimethyl sulfoxide solution to each well, and shake on an oscillator at an appropriate speed for 5 min to dissolve the purple crystals. Observe the color change, place the 96-well plate in an ELISA reader, detect the absorbance (OD) value of each well at a wavelength of 490 nm, and compare it with the blank group.

[0367] (4) Data analysis: Calculate the cell proliferation rate and inhibition rate of each group.

[0368] Cell survival rate = (average OD value of the test group / OD value of the blank group) × 100%

[0369] Inhibition rate = (1-average OD value of the test group / OD value of the blank group) × 100%

[0370] Calculate the half effective inhibitory concentration (IC 50 ): Based on the drug concentration and the growth inhibition rate of the drug on cells, the IC 50 value.

[0371] Compounds inhibit human lung cancer cell line H460 WT and H460 R231Q The cell activity results are shown in the table below, where "A" represents IC 50 Values ​​< 10 μM; "B" means 10 μM < IC 50 Values ​​< 100 μM; “C” indicates IC 50 Value>100μM.

[0372] Table 1 Compounds inhibit human lung cancer cell line H460 WT and H460 R231Q Cell viability results

[0373]

[0374]

[0375] 2. In vitro enzyme inhibition activity

[0376] Mix 2 μL enzyme solution, 2 μL substrate (20 μM SAM, 0.15 ng / mL Nuclesomes) and 1 μL test compound or positive control compound (SAH). The control group includes all substrates except the test compound. The total volume of DOT1L test reaction solution is 5 μL. After dilution with Tris buffer (20 mM Tris-HCl, 0.01% Trixton-100, 0.1% BSA, 0.5 mM DTT, pH = 8.0), incubate at room temperature for 3 hours, add 2.5 μL high salt buffer (50 mM Tris-HCl, 1 M NaCl, 0.1% Tween-20, 0.3% poly-L-lysine, pH = 7.4) to terminate the reaction, detect with the detection kit, and use Envision to measure the fluorescence intensity at an excitation wavelength of 680 nM and an emission wavelength of 615 nM. Three replicates were set for all doses, and nonlinear fitting was performed using GraphPadPrism 5 to calculate IC 50 value.

[0377] Some compounds of the present invention inhibit DOT1L WT The enzyme activity results are shown in Table 2, in which "A" represents IC 50 Value < 100nM; "B" means 100nM < IC 50 Value < 1 μM; "C" means 1 μM < IC 50 Value < 10 μM; "D" means 10 μM < IC 50 Values ​​<100 μM.

[0378] Table 2 Compounds inhibit DOT1L WT Enzyme activity results

[0379]

Claims

1. A 1,3-disubstituted tetrahydropyrrole derivative, characterized in that: The chemical structural formula of the 1,3-disubstituted tetrahydropyrrole derivative is shown in the general formula (I) or (II): Where: Het is an 8-10 membered heteroaryl substituted by 0-4 independent R1, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; R1 is halogen, hydroxyl, amino, nitro, cyano, trifluoromethyl, trifluoromethoxy, (C1-C6) alkyl, (C1-C6) alkoxy, (C1-C6) acylamino, or amino substituted by 1-2 (C1-C6) alkyl groups; R2 is hydrogen, (C1-C6) alkyl, (C1-C6) acyl; R3 is hydrogen, (C1-C6) alkyl, (C2-C6) alkenyl, (C2-C6) alkynyl, halogen or amino or cyano or carboxyl substituted (C1-C6) alkyl, (CH2) p (C3-C6)cycloalkyl, (C1-C6)acyl, (C1-C6)sulfonyl, (CH2) p NR5R6, (CH2) p CONR5R6, (CH2) p COOR5, (CH2) p OR5; R5 and R6 are the same or different and are independently selected from hydrogen, (C1-C6) alkyl and halogen or amino or hydroxy or cyano substituted (C1-C6) alkyl; or R5 and R6 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic or heteroaryl group, wherein the heterocyclic or heteroaryl group contains 0-4 heteroatoms selected from N, O and S in addition to the nitrogen atom to which R5 and R6 are attached; X is methylene, (CH2) m CONH(CH2) n 、(CH2) m NH(CH2) n 、(CH2) m COO(CH2) n 、(CH2) m O(CH2) n ; Ar is a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and S; R4 represents 0-4 substituents independently selected from halogen, hydroxy, amino, nitro, cyano, (C1-C6) alkyl, (C1-C6) alkoxy, (C3-C6) cycloalkyl, (C1-C6) alkoxycarbonyl, (C1-C6) alkyl substituted with 0-3 halogen, hydroxy or amino, (C1-C6) alkoxy substituted with 0-3 halogen, hydroxy or amino, (C1-C6) acyl substituted with 0-3 halogen, hydroxy or amino, (C1-C6) acylamino substituted with 0-3 halogen, hydroxy or amino, and (C1-C6) sulfonylamino substituted with 0-3 halogen, hydroxy or amino; p is independently 0-6; m is independently 0-3; n is independently 0-3.

2. A 1,3-disubstituted tetrahydropyrrole derivative according to claim 1, characterized in that: In the general formula (I) or (II): Het is an 8-10 membered heteroaryl substituted by 0-4 independent R1, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; R1 is halogen, hydroxy, amino, nitro, cyano, trifluoromethyl, trifluoromethoxy, (C1-C3) alkyl, (C1-C3) alkoxy, (C1-C3) acylamino, or amino substituted by 1-2 (C1-C3) alkyl groups; R2 is hydrogen, (C1-C3) alkyl or (C1-C3) acyl; R3 is hydrogen, (C1-C3) alkyl, (C2-C4) alkenyl, (C2-C4) alkynyl, halogen or amino or cyano or carboxyl substituted (C1-C3) alkyl, (CH2) p (C3-C6)cycloalkyl, (C1-C3)acyl, (C1-C3)sulfonyl, (CH2) p NR5R6, (CH2) p CONR5R6, (CH2) p COOR5 or (CH2) p OR5; R5 and R6 are the same or different and are independently selected from hydrogen, (C1-C3) alkyl and halogen or amino or hydroxy or cyano substituted (C1-C3) alkyl; or R5 and R6 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic or heteroaryl group, wherein the heterocyclic or heteroaryl group contains 0-4 heteroatoms selected from N, O and S in addition to the nitrogen atom to which R5 and R6 are attached; X is methylene, (CH2) m CONH(CH2) n 、(CH2) m NH(CH2) n 、(CH2) m COO(CH2) n 、(CH2) m O(CH2) n ; Ar is a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and S; R4 represents 0-3 substituents independently selected from halogen, hydroxy, amino, nitro, cyano, (C1-C6) alkyl, (C1-C6) alkoxy, (C3-C6) cycloalkyl, (C1-C6) alkoxycarbonyl, (C1-C6) alkyl substituted with 0-3 halogen, hydroxy or amino, (C1-C6) alkoxy substituted with 0-3 halogen, hydroxy or amino, (C1-C6) acyl substituted with 0-3 halogen, hydroxy or amino, and (C1-C6) acylamino substituted with 0-3 halogen, hydroxy or amino, or (C1-C6) sulfonylamino substituted with 0-3 halogen, hydroxy or amino; p is independently 0-4; m is independently 0-2; n is independently 0-2.

3. A 1,3-disubstituted tetrahydropyrrole derivative according to claim 2, characterized in that: In the general formula (I) or (II): Het is an 8-10 membered heteroaryl substituted by 0-2 independent R1, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; R1 is halogen, hydroxy, amino, nitro, cyano, trifluoromethyl, trifluoromethoxy, (C1-C3) alkyl, (C1-C3) acylamino, or amino substituted by 1-2 (C1-C3) alkyl groups; R2 is hydrogen, (C1-C3) alkyl, (C1-C3) acyl; R3 is hydrogen, (C1-C3) alkyl, halogen or amino or cyano or carboxyl substituted (C1-C3) alkyl, (C1-C3) acyl, (CH2) p NR5R6, (CH2) p CONR5R6, (CH2) p COOR5, (CH2) p OR5; R5 and R6 are the same or different and are independently selected from hydrogen and (C1-C3) alkyl; or R5 and R6 together with the nitrogen atom to which they are attached form a 4-10 membered heterocyclic or heteroaryl group, wherein the heterocyclic or heteroaryl group contains 0-4 heteroatoms selected from N, O and S in addition to the nitrogen atom to which R5 and R6 are attached; X is methylene, (CH2) m CONH(CH2) n 、(CH2) m NH(CH2) n ; Ar is a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and S; R4 represents 0-2 substituents independently selected from halogen, hydroxy, amino, nitro, cyano, (C1-C6) alkyl, (C1-C6) alkoxy, (C1-C6) alkoxycarbonyl, (C1-C6) alkyl substituted with 0-3 halogen, hydroxy or amino, (C1-C6) alkoxy substituted with 0-3 halogen, hydroxy or amino, (C1-C6) acyl substituted with 0-3 halogen, hydroxy or amino, (C1-C6) acylamino substituted with 0-3 halogen, hydroxy or amino, and (C1-C6) sulfonylamino substituted with 0-3 halogen, hydroxy or amino; p is independently 0-4; m is independently 0-2; n is independently 0-2.

4. A 1,3-disubstituted tetrahydropyrrole derivative according to claim 3, characterized in that: In the general formula (I) or (II): Het is a 9-membered heteroaryl substituted by 0-2 independent R1, wherein the heteroaryl contains 1-4 heteroatoms selected from N, O and S; R1 is halogen, hydroxy, amino, nitro, cyano, trifluoromethyl, trifluoromethoxy or (C1-C3) alkyl; R2 is hydrogen or (C1-C3) acyl; R3 is hydrogen, (C1-C3) alkyl, (CH2) p NR5R6, (CH2) p CONR5R6 or (CH2) p COOR5; R5 and R6 are the same or different and are independently selected from hydrogen and (C1-C3) alkyl; or R5 and R6 together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclic group, which contains 0-4 heteroatoms selected from N, O and S in addition to the nitrogen atom to which R5 and R6 are attached; X is methylene or (CH2) m CONH(CH2) n ; Ar is a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and S; R4 represents 0-1 substituents independently selected from (C1-C6) alkyl and (C1-C6) alkoxycarbonyl or 0-3 halogen or hydroxy or amino substituted (C1-C6) acylamino groups; p is independently 0-4; m is independently 0-1; n is independently 0-1.

5. A 1,3-disubstituted tetrahydropyrrole derivative according to claim 4, characterized in that: In the general formula (I) or (II): Het is a 9-membered heteroaryl substituted by 0-2 R1, wherein the heteroaryl is optionally pyrazolo[3,4-d]pyrimidine, imidazo[1,2-a]pyrazine, pyrrolo[2,3-d]pyrimidine or purine; R1 is halogen, amino or methyl; R2 is hydrogen or acetyl; R3 is hydrogen, X is Ar is a benzene ring, piperidine or thiophene; R4 represents 0-1 substituents independently selected from tert-butyl and methoxycarbonyl or pivalylamide.

6. A 1,3-disubstituted tetrahydropyrrole derivative according to any one of claims 1 to 5, characterized in that: Selected from the following compounds: 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(isopropyl)amino}propyl}urea; 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(isopropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea; 1-[4-(tert-butyl)phenyl]-3-{3-{{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}(3-morpholinopropyl)amino}propyl}urea; 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholinopropyl)amino}propyl}urea; 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholinopropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea; 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholino-3-oxopropyl)amino}propyl}urea; 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-morpholino-3-oxopropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea; 1-[4-(tert-butyl)phenyl]-3-{3-{{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}(3-morpholino-3-oxopropyl)amino}propyl}urea; 1-[4-(tert-butyl)phenyl]-3-{3-{{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}urea; 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}urea; 1-[4-(tert-butyl)phenyl]-3-{3-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}urea; 1-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(3-oxo-3-thiomorpholinylpropyl)amino}propyl}-3-[4-(tert-butyl)phenyl]urea; Methyl 3-{{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}{[3-hydroxy-1-(9H-purin-6-yl)pyrrolidin-3-yl]methyl}amino}propanoate; Methyl 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}amino}propanoate; Methyl 3-{{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanoate; Methyl 3-{{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanoate; Methyl 3-{{[3-acetoxy-1-(2-chloro-9H-purin-6-yl)pyrrolidin-3-yl]methyl}{3-{3-[4-(tert-butyl)phenyl]ureido}propyl}amino}propanoate; N-{2-{{4-{{[3-hydroxy-1-(9H-purin-6-yl)]pyrrolidin-3-yl}methyl(methyl)amino}piperidin-1-yl}methyl}-1H-benzo[d]imidazol-5-yl}pivalamide; N-{2-{{4-{{[3-hydroxy-1-(2-chloro-9H-purin-6-yl)]pyrrolidin-3-yl}methyl(methyl)amino}piperidin-1-yl}methyl}-1H-benzo[d]imidazol-5-yl}pivalamide; N-{2-{{4-{{[3-hydroxy-1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)]pyrrolidin-3-yl}methyl(methyl)amino}piperidin-1-yl}methyl}-1H-benzo[d]imidazol-5-yl}pivalamide; 2-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide; N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}acetamide; N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}acetamide; 2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide; 3-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide; N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}propanamide; N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}propanamide; 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide; 3-{3-{2-{2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}acetamide}ethyl}ureido}thiophene-2-carboxylic acid methyl ester; 3-{3-{2-{3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}propionamide}ethyl}ureido}thiophene-2-carboxylic acid methyl ester; 2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(pyridin-3-yl)ureido]ethyl}acetamide; 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}(methyl)amino}-N-{2-[3-(pyridin-3-yl)ureido]ethyl}propanamide; 2-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide; N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}acetamide; N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-2-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}acetamide; 2-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}acetamide; 3-{{[1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide; N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(2-chloro-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanamide; N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(6-chloro-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanamide; 3-{{[1-(3-bromoimidazo[1,2-a]pyrazin-8-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide; N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}-3-{{[1-(2-chloro-9-methyl-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}propanamide; 3-{{[1-(2-amino-9-methyl-9H-purin-6-yl)-3-hydroxypyrrolidin-3-yl]methyl}amino}-N-{2-[3-(4-tert-butylphenyl)ureido]ethyl}propanamide.

7. A method for preparing a 1,3-disubstituted tetrahydropyrrole derivative according to any one of claims 1 to 5, characterized in that: The preparation method is selected from one of method A, method B and method C (1) Method A: Step 1: Compound 1-tert-butyloxycarbonyl-3-pyrrolidone undergoes a nucleophilic addition reaction with nitromethane under alkaline conditions to generate intermediate M1; Step 2: Intermediate M1 undergoes reduction reaction in the presence of a reducing agent to generate intermediate M2; Step 3: Different substituted aryl or heteroaryl primary amines react with ethylenediamine in the presence of carbonyldiimidazole (CDI) to generate intermediate M4; Step 4: In the presence of a condensing agent, the intermediate M4 undergoes a condensation reaction with bromoacetic acid to generate an intermediate M5; or the intermediate M4 undergoes a condensation reaction with acrylic acid to generate an intermediate M6; Step 5: Intermediate M2 and intermediate M3 undergo a reductive amination reaction in the presence of a reducing agent to generate intermediate M7; or intermediate M5 and intermediate M2 undergo an N-alkylation reaction under alkaline conditions to generate intermediate M7; or intermediate M5 and intermediate M2 undergo a Michael addition reaction to generate intermediate M7; Step 6: Intermediate M7 is subjected to reductive amination reaction, Michael addition reaction or N-alkylation reaction in the presence of a base to generate intermediate M8; Step 7: The intermediate M8 is deprotected from tert-butyloxycarbonyl (Boc) under acidic conditions to generate intermediate M9; Step 8: Intermediate M9 undergoes N-alkylation reaction with differently substituted heteroaryl halides under alkaline conditions to prepare compound I; Method B: Step 1: The intermediate M1 is deprotected from tert-butyloxycarbonyl (Boc) under acidic conditions to generate intermediate M10; Step 2: Intermediate M10 undergoes N-alkylation reaction with heteroaryl halides of different substitutions under alkaline conditions to generate intermediate M11; Step 3: Intermediate M11 undergoes reduction reaction in the presence of a reducing agent to generate intermediate M12; Step 4: Intermediate M12 and intermediate M5 undergo N-alkylation reaction under alkaline conditions to obtain a compound represented by general formula (I); or intermediate M5 and intermediate M2 undergo Michael addition reaction to obtain compound I; Method C: Step 1: Intermediate M13 and intermediate M2 undergo reductive amination reaction in the presence of a reducing agent to generate intermediate M14; Step 2: Intermediate M14 undergoes reductive amination reaction with various aldehyde-containing compounds in the presence of a reducing agent to generate intermediate M15; Step 3: The intermediate M15 is deprotected from the tert-butyloxycarbonyl (Boc) group under acidic conditions to generate the intermediate M16; Step 4: Intermediate M16 undergoes N-alkylation reaction with heteroaryl halides of different substitutions under alkaline conditions to obtain compound II; The substituents Het, R1, R2, R3, R4, X and Ar involved in Method A, Method B and Method C are as defined in Claims 1-5.

8. Use of a 1,3-disubstituted tetrahydropyrrole derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 in the preparation of a DOT1L inhibitor.

9. Use of a 1,3-disubstituted tetrahydropyrrole derivative or a pharmaceutically acceptable salt thereof for preparing a DOT1L inhibitor according to claim 8, characterized in that: The DOT1L inhibitor is used for preparing drugs for treating tumors.

10. A pharmaceutical composition comprising a 1,3-disubstituted tetrahydropyrrole derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, and a pharmaceutically acceptable excipient.

11. Use of the pharmaceutical composition according to claim 10 in the preparation of a DOT1L inhibitor.

12. Use of a pharmaceutical composition according to claim 11 in the preparation of a DOT1L inhibitor, characterized in that: The DOT1L inhibitor is used for preparing drugs for treating tumors.

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