Aniline-based WDR5 protein-protein interaction inhibitor, preparation method therefor and use thereof

Abstract

The present disclosure discloses a WDR5 protein-protein interaction inhibitor, including a compound having a structure represented by general formula (I). Experiments show that the inhibitor acts on a WDR5 protein and an interacting protein thereof including, but not limited to, MLL, selectively inhibits the proliferation of leukemia cells, and inhibits the methylation of H3K4 and the expression of downstream Hox / Meis-1 gene at the cellular level. The present disclosure also discloses a method for preparing the inhibitor and use thereof in the preparation of a drug for treating acute leukemia and other related diseases.

Description

Anilines as wdr5 protein-protein interaction inhibitors and methods of making and using the same TECHNICAL FIELD

[0001] The present application relates to an aniline compound and methods of making and using the same in the field of medicinal chemistry. The compound as WDR5 protein-protein interaction inhibitor can inhibit the expression of histone methyltransferase and downstream hematopoietic genes at the cellular level, and inhibit the growth of xenograft tumors in nude mice at the animal level, and can be applied to treat diseases such as acute leukemia. BACKGROUND

[0002] WDR5 protein is composed of 334 amino acids, containing 7 WD40 repeat domains. Structural studies have shown that WD40 repeat structure forms a 7-leaf propeller structure, and each leaf propeller contains 4 anti-parallel chains. Such structural characteristics enable WDR5 to interact with many other proteins, such as WDR5-MLL1, WDR5-MYC, and WDR5-MOF / HAT. WDR5 can also bind to H3K4me2. Studies have shown that WDR5 can stabilize or assemble virus-induced signaling adaptor (VISA)-related complexes, and plays an important role in type I interferon and antiviral innate immune responses. WDR5 protein is also highly expressed in bladder cancer, promotes the proliferation, self-renewal and chemotherapy of bladder cancer cells by activating the expression of H3K4me3, and participates in N-Myc-regulated transcriptional activation and tumor generation as an important cofactor. Inhibition of WDR5 protein serves as a new target for treating MYCN-amplified neuroblastoma.

[0003] Histone methylation modification plays a key role in many biological processes and is an important research content in the field of epigenetic regulation. MLL1 is a histone H3 lysine 4 (H3K4) methyltransferase. MLL1 gene translocation rearrangement forms a fusion gene, which expresses MLL fusion protein with carcinogenic effect, thereby inducing mixed lineage leukemia (MLL1, acute myeloid leukemia and acute lymphoblastic leukemia). The treatment of leukemia caused by MLL abnormalities is not ideal by conventional chemotherapy, and the prognosis is poor. There is currently no targeted drug, so it is urgent to develop new therapeutic drugs targeting the biological properties of the disease.

[0004] Chromosomal translocation of MLL1 only occurs on a single allele, and there is also a wild-type MLL1. Wild-type MLL1 and its fusion protein are involved in the occurrence and development of mixed lineage leukemia, and the enzyme catalytic activity of wild-type MLL1 is essential for the generation of leukemia induced by MLL fusion protein. Therefore, specifically inhibiting the enzyme catalytic activity of wild-type MLL1 can achieve the effect of treating leukemia.

[0005] MLL1 alone has poor catalytic activity and can only catalyze H3K4 monomethylation; when MLL1 forms a core complex with WDR5, RbBP5, Ash2L and DPY30, the catalytic activity of the enzyme is greatly improved, especially for H3K4me2. Among them, WDR5 plays a crucial role in the formation of the complex as a connecting bridge connecting the C-terminal WIN motif part of MLL1 with other proteins. Therefore, interfering with the protein-protein interaction of WDR5 using a small molecule inhibitor is an effective means to inhibit the catalytic activity of MLL1, thereby inhibiting the expression of downstream Hox and Meis-1 genes, and further blocking the progression of leukemia.

[0006] SUMMARY

[0007] The purpose of the present application is to provide a WDR5 protein-protein interaction inhibitor, which can inhibit the expression of histone methyltransferase and downstream hematopoietic genes at the cellular level, inhibit the growth of nude mouse xenografts at the animal level, and be applied in the treatment of indications related to MLL enzyme function. It can be applied to the treatment of acute leukemia and other diseases.

[0008] Technical scheme: The present application discloses a small molecule compound which can interfere with the protein-protein interaction of WDR5. By interfering with the protein-protein interaction of WDR5, the methyltransferase catalytic activity of MLL1 is inhibited, the expression level of Hox and Meis-1 genes is down-regulated, thereby inducing the apoptosis of leukemia cells, and the compound is used for treating acute leukemia. The structure of the compound of the present application is as follows:

[0009]

[0010] X represents CH or N;

[0011] Y represents C or N;

[0012] R 1 , R 2 and R 3 are the same or different from each other and are each independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, nitro, halogen, cyano, aldehyde, hydroxyl, -NR 7 R 8 , or R 2 , R 3 are linked to form a wherein R 7 , R 8 are the same or different from each other and are each independently hydrogen or C1-C4 alkyl, or R 7 , R 8 are linked to form a 3-7 membered nitrogen-containing heterocyclic ring;

[0013] R 4 represents morpholinyl, piperazinyl, 4-substituted piperazinyl, 4-substituted homopiperazinyl, 3-substituted piperazinyl or 2-substituted piperazinyl, the substituents being C1-C4 alkyl, 3-7 membered cycloalkyl, hydroxyalkyl, phenyl;

[0014] R 5 represents nitro, amino, phenyl, substituted phenyl, 5-6 membered aromatic heterocycle containing oxygen or nitrogen, substituted 5-6 membered aromatic heterocycle containing oxygen or nitrogen, -NHCOR 9 ; wherein R 9 represents hydroxy, C1-C6 alkoxy, phenyl, substituted phenyl, 5-6 membered aromatic heterocycle containing oxygen or nitrogen, substituted 5-6 membered aromatic heterocycle containing oxygen or nitrogen, the substituents being C1-C4 alkyl, C1-C4 alkoxy, halogen, cyano, -NHCOR 10 , -CONR 11 R 12 or -COOR 10 , wherein R 10 represents hydrogen, C1-C6 alkyl, C1-C6 amino-substituted alkyl, 3-7 membered cycloalkyl, 3-7 membered heterocycle containing nitrogen or oxygen, phenyl; R 11 , R 12 independently represent hydrogen, C1-C6 alkyl, phenyl or substituted phenyl, substituted or unsubstituted 3-7 membered heterocycle containing nitrogen or oxygen or R 11 , R 12 are linked to form a 3-7 membered heterocycle containing nitrogen or oxygen;

[0015] R 6 represents hydrogen, halogen, methyl, trifluoromethyl, amino, substituted amino, the substituents being C1-C4 alkyl, allyl;

[0016] R 1 , R 2 and R 3 are the same as or different from each other and are each independently preferably selected from the group consisting of hydrogen, halogen, methyl, nitro, cyano, aldehyde, methoxy, -NR 7 R 8 , or R 2 , R 3 are linked to form a wherein R 7 , R 8 are the same as or different from each other and are each independently hydrogen or C1-C4 alkyl or R 7 , R 8 are linked to form a 3-5 membered nitrogen-containing heterocycle;

[0017] R 4 is preferably morpholinyl, piperazinyl, 4-substituted piperazinyl, the substituents being methyl, ethyl, cyclopropyl, hydroxyethyl, phenyl;

[0018] R 5 Preferably, the substituents are nitro, amino, -NHCO R 9 , furanyl, pyrimidinyl, pyridinyl, substituted triazole, substituted phenyl, the substituents being mono- or di-substituted halogen, -NHCOR 10 , -CONR 11 R 12 or -COOR 10 ; R 9 represents phenyl; R 10 represents hydrogen, C1-C6 alkyl, C1-C6 amino-substituted alkyl, 3-7 membered cycloalkyl, 3-7 membered nitrogen- or oxygen-containing heterocycle, phenyl; R 11 , R 12 independently represent hydrogen, C1-C6 alkyl, phenyl or substituted phenyl, substituted or unsubstituted 3-7 membered nitrogen- or oxygen-containing heterocycle or R 11 , R 12 are linked to form a 3-7 membered nitrogen- or oxygen-containing heterocycle.

[0019] The present application also includes pharmaceutically acceptable salts of the compound (I), and solvates thereof, all of which have the same pharmacological effects as the compound (I).

[0020] The present application discloses a pharmaceutical composition comprising the compound (I) or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable carriers, diluents and excipients.

[0021] The present application also provides the use of the compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof in the preparation of a medicament for treating a disease mediated by the enzyme by inhibiting the protein-protein interaction of WDR5, such as MLL gene fusion leukemia, which can be treated by inhibiting the enzymatic activity of MLL1.

[0022] The dosage of the compound of the present application for clinical use is 0.01 mg to 1000 mg per day, which can deviate from the range according to the severity of the disease or the different dosage forms.

[0023] In some embodiments, the compound according to formula (I) can contain basic functional groups sufficient to form salts. Representative salts include pharmaceutically acceptable inorganic acid salts, such as hydrochloride, hydrobromide and sulfate; pharmaceutically acceptable organic acid salts, preferably acetate, trifluoroacetate, lactate, succinate, fumarate, maleate, citrate, benzoate, methanesulfonate, p-toluene sulfonate and p-toluene sulfonic acid salt.

[0024] The present application also discloses a preparation method of the compound of formula (I), which comprises the following method:

[0025]

[0026]

[0027] Beneficial effects: the aniline compound of the present application has strong WDR5 protein-protein interaction inhibition activity, can reduce MLL1 enzyme catalytic activity at the cellular level, down-regulate Hox and Meis-1 gene expression, and treat indications related to WDR5 enzyme function, which are blood tumors, including acute leukemia and other diseases. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 is a Western-blot experiment to detect the effect of Example 72 on MLL1 enzyme catalytic activity in cells;

[0029] Figure 2 is an RT-PCR experiment to detect the effect of Example 72 on down-regulating Hoxa9 and Meis-1 gene expression in cells;

[0030] Figure 3 is a MV4-11 nude mouse xenograft model to detect the inhibition level of Example 72 on tumors at the animal level. DETAILED DESCRIPTION

[0031] Example 1

[0032]

[0033] N-(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)quinazolin-4-amine

[0034] Step 1: Preparation of 2-(4-methylpiperazin-1-yl)-5-nitroaniline

[0035] 2-fluoro-5-nitroaniline (2 g, 12.8 mmol) was dissolved in DMF (20 mL), DIPEA (3.3 g, 25.6 mmol) and 1-methylpiperazine (25.6 mmol) were added, and the reaction was stirred at 100°C for 8 h. 100 mL of water was added to the reaction solution, extracted with ethyl acetate (3 x 50 mL), the organic layers were combined and dried over anhydrous Na2SO4, and the crude product was obtained after distillation under reduced pressure, washed with CH3CN to obtain a yellow solid, with a yield of 53.0%; m.p. 200-202°C; 1 H NMR (300 MHz, DMSO-d6) δ 7.53 (d, J = 2.4 Hz, 1H), 7.42 (dd, J = 8.8, 2.4 Hz, 1H), 7.04 (d, J = 8.7 Hz, 1H), 5.26 (s, 2H), 3.11 (s, 4H), 2.58 (s, 4H), 2.24 (s, 3H). HRMS (ESI): calcd. for m / z C 11 H 17 N4O2, [M+H]+ 237.1316, found 237.1342.

[0036] Step 2: Preparation of the title compound N-(2-(4-methylpiperazin- 1 -yl)-5- nitrophenyl)quinazolin-4-amine

[0037] Dissolve 2-(4-methylpiperazin-l-yl)-5-nitroaniline (0.2 g, 0.846 mmol) in isopropanol (10 mL), add 1 mL of concentrated HCl and 4-chloroquinazoline (0.27 g, 1.69 mmol) and reflux for 18 h, distill under reduced pressure until solid precipitates, cool, suction filter, wash the filter cake with isopropanol (5 mL x 3) and dry to give a yellow solid in 63.5% yield; m.p. > 250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.57 (s, 1H), 8.61 (s, 2H), 8.43 (s, 1H), 8.07 (s, 1H), 7.86 (d, J = 10.0 Hz, 2H), 7.69 (s, 1H), 7.30 (s, J = 8.8 Hz, 1H), 3.09 (s, 4H), 2.32 (s, 4H), 2.13 (s, 3H). HRMS (ESI): calcd. for m / z C 19 H 20 N6O2, [M+H] + 365.1720, found 365.1722. HPLC (100% MeOH): t R = 7.772 min, 99.16%.

[0038] Example 2

[0039]

[0040] 6-chloro-N-(2-(4-methylpiperazin-l-yl)-5-nitrophenyl)pyrimidin-4-amine

[0041] Following the procedure of Example 1, Step 2, replace 4-chloroquinazoline with 4,6-dichloropyrimidine (1.69 mmol) to give a yellow solid in 65.8% yield; m.p. 169 °C - 171 °C; 1H NMR (300MHz, DMSO-d6) δ9.36(s,1H),8.48(s,1H),8.42(d,J=2.7Hz,1H),8.02(dd,J=8.9,2.8Hz,1H),7.23(d ,J=9.0Hz,1H),6.81(s,1H),3.04(t,J=4.6Hz,4H),2.35(t,J=4.7Hz,4H),2.16(s,3H).HRMS(ESI):calcd.for m / z C 15 H 17 ClN6O2,[M+H] + 349.1174,found 349.1177.HPLC(100%MeOH):t R =6.097 min, 96.84%.

[0042] Example 3

[0043]

[0044] N 4 N 4 -dimethyl-N 6 -(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)pyrimidin-4,6-diamine

[0045] Example 2 (0.35 mmol) was dissolved in DMF (5 mL), and DIPEA (1.5 mmol) and dimethylamine (1 mmol) were added. The mixture was reacted at 120 °C for 4 h. The reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL × 3), washed with saturated sodium chloride, and the organic layer was dried over anhydrous sodium sulfate. The mixture was filtered, evaporated to dryness, and the crude product was subjected to silica gel column chromatography (dichloromethane:methanol = 50:1) to give a yellow solid. Yield 49.5%; mp 152 °C-154 °C; 1 H NMR (300MHz, CDCl3) δ8.69(d,J=2.7Hz,1H),8.28(s,1H),7.80(dd,J=8.7,2.7Hz,1H),7.06(d,J=8.8Hz,1H),6.9 4(s,1H),5.82(s,1H),3.03(s,6H),2.94(t,J=4.8Hz,4H),2.65-2.48(m,4H),2.30(s,3H).HRMS(ESI):calcd.for m / z C 17 H 23 N7O2,[M+H] + 358.1986,found 358.1992.HPLC(100%MeOH):tR = 4.709 min, 99.20 %.

[0046] Example 4

[0047]

[0048] 6-Chloro-N-(2-(4-methylpiperazin- 1 -yl)-5-nitrophenyl)pyrimidin-4- amine

[0049] Following the procedure of Example 1, Step 2, using 4,6-dichloro-2- methylpyrimidine in place of 4-chloroquinazoline gave a yellow solid in 44.8% yield; m.p. 196-198 °C; 1 H NMR (300 MHz, CDC13) δ 9.09 (d, J = 2.6 Hz, 1H), 7.97 (dd, J = 8.8, 2.7 Hz, 1H), 7.55 (s, 1H), 7.23 (s, 1H), 6.59 (s, 1H), 3.02 (t, J = 4.8 Hz, 4H), 2.65-2.61 (m, 7H), 2.40 (s, 3H). HRMS (ESI): calcd. for m / z C 16 H 19 ClN6O2, [M+H] + 363.1331, found 363.1335. HPLC (100% MeOH): t R = 8.574 min, 97.99 %.

[0050] Example 5

[0051]

[0052] 6-Chloro-N 4 -(2-(4-methylpiperazin- 1 -yl)-5-nitrophenyl)pyrimidin-2,4-diamine

[0053] Following the procedure of Example 1, Step 2, using 4,6-dichloro-2- methylpyrimidine in place of 4-chloroquinazoline gave a yellow solid in 44.8% yield; m.p. 196-198 °C; 1 H NMR (300 MHz, CDC13) δ 9.09 (d, J = 2.6 Hz, 1H), 7.97 (dd, J = 8.8, 2.7 Hz, 1H), 7.55 (s, 1H), 7.23 (s, 1H), 6.59 (s, 1H), 3.02 (t, J = 4.8 Hz, 4H), 2.65-2.61 (m, 7H), 2.40 (s, 3H). HRMS (ESI): calcd. for m / z C 15 H 18CIN7O2, [M+H] + 364.1283, found 364.1289. HPLC (100% MeOH): t R = 5.850 min, 97.14%.

[0054] Example 6

[0055]

[0056] 6-chloro-5-methyl-N-(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)pyrimidin-4- amine

[0057] Following the preparation procedure of Example 1, Step 2, using 4,6-dichloro-5- methylpyrimidine in place of 4-chloroquinazoline gave a yellow solid in 60.5% yield; m.p. 259-261 °C; 1 H NMR (300 MHz, CDC13) δ 9.55 (s, 1H), 8.57 (s, 1H), 8.18 (s, 1H), 7.95 (dd, J = 8.7, 2.7 Hz, 1H), 7.27 (s, 1H), 3.02 (t, J = 4.8 Hz, 4H), 2.66 (s, 4H), 2.42 (s, 6H). HRMS (ESI): calcd. for m / z C 16 H 19 CIN6O2, [M+H] + 363.1331, found 363.1349. HPLC (100% MeOH): t R = 7.673 min, 97.05%.

[0058] Example 7

[0059]

[0060] 6-chloro-N 4 -(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)pyrimidine-4,5-diamine

[0061] Following the preparation procedure of Example 1, Step 2, using 4,6-dichloro-5- methylpyrimidine in place of 4-chloroquinazoline gave a yellow solid in 60.5% yield; m.p. 259-261 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.65 (d, J = 2.6 Hz, 1H), 8.22 (s, 1H), 7.98 - 7.91 (m, 2H), 7.27 (d, J = 8.9 Hz, 1H), 5.41 (s, 2H), 2.91 (br s, 4H), 2.47 (br s, 4H), 2.20 (s, 3H). HRMS (ESI): calcd. for m / z C 15 H 18 ClN7O2, [M+H] + 364.1283, found 364.1286. HPLC (100% MeOH): t R = 6.581 min, 98.22%.

[0062] Example 8

[0063]

[0064] 6-chloro-2-methyl-N 4 -(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)pyrimidine-4,5-diamine

[0065] Following the preparation procedure of Example 1, Step 2, using 4,6-dichloro 2-methyl-5- aminopyrimidine in place of 4-chloroquinazoline to give a yellow solid in 67.8% yield; m.p. 248-250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.87 (d, J = 2.8 Hz, 1H), 8.22 (s, 1H), 7.94 (dd, J = 8.8, 2.8 Hz, 1H), 5.12 (s, 2H), 2.98 (t, J = 4.8 Hz, 4H), 2.46 (s, 4H), 2.32 (s, 3H), 2.21 (s, 3H). HRMS (ESI): calcd. for m / z C 16 H 20 ClN7O2, [M+H] + 378.1440, found 378.1440. HPLC (100% MeOH): t R = 6.251 min, 97.99%.

[0066] Example 9

[0067]

[0068] N 4 -(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)pyrimidine-4,5-diamine

[0069] Preparation according to the procedure of Example 1, Step 2, using 4-chloro-5- aminopyrimidine instead of 4-chloroquinazoline to give a yellow solid in 43.7% yield; m.p. >250°C; 1 H NMR (300 MHz, DMSO-d6) δ 8.25 (s, 1H), 7.82 (dd, J = 7.4, 1.9 Hz, 1H), 7.81-7.77 (m, 2H), 7.67 (s, 1H), 7.04 (d, J = 7.5 Hz, 1H), 5.82 (s, 2H), 3.20 (t, J = 5.3 Hz, 4H), 2.98 (t, J = 5.3 Hz, 4H), 2.60 (s, 3H). HRMS (ESI): calcd. for m / z C 15 H 19 N7O2, [M+H] + 330.1673, found 330.1675. HPLC (100% MeOH): t R = 7.673 min, 97.05%

[0070] Example 10

[0071]

[0072] N-(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)pyrimidin-2-amine

[0073] Preparation according to the procedure of Example 1, Step 2, using 2-bromopyrimidine instead of 4-chloroquinazoline to give a yellow solid in 57.5% yield; m.p. 195°C-197°C; 1 H NMR (300 MHz, CDCl3) δ 9.47 (s, 1H), 8.54 (d, J = 3.0 Hz, 2H), 8.03 (s, 1H), 7.88 (d, J = 3.0 Hz, 1H), 7.22 (d, J = 9.0 Hz, 1H), 6.85 (t, J = 9.0 Hz, 1H), 3.04 (t, J = 5.6 Hz, 4H), 2.70 (br s, 4H), 2.42 (s, 3H). HRMS (ESI): calcd. for m / z C 15 H 18 ClN7O2, [M+H] + 315.1564, found 315.1566. HPLC (100% MeOH): t R = 16.340 min, 96.09%.

[0074] Example 11

[0075]

[0076] N 2 -(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)pyrimidine-2,4-diamine

[0077] Following the procedure of Example 1, Step 2, replacing 4-chloroquinazoline with 2-chloropyrimidin-4-amine gave a yellow solid in 45.7% yield; m.p. 210-212 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.30 (s, 1H), 7.94 (d, J = 6.0 Hz, 1H), 7.80 (dd, J = 8.7 Hz, 1H), 7.63 (s, 1H), 7.32 (d, J = 8.8 Hz, 1H), 6.81 (br s, 2H), 6.03 (d, J = 5.8 Hz, 1H), 2.93 (t, J = 4.7 Hz, 4H), 2.54 (br s, 4H), 2.26 (s, 3H). HRMS (ESI): calcd. for m / z C 15 H 19 N7O2, [M+H] + 330.1673, found 330.1679. HPLC (100% MeOH): t R = 6.354 min, 96.85%.

[0078] Example 12

[0079]

[0080] N 2 -(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)pyrimidine-2,4-diamine

[0081] Following the procedure of Example 1, Step 2, replacing 4-chloroquinazoline with 2-chloropyridin-3-amine gave a yellow solid in 36.6% yield; m.p. 210-212 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.06 (d, J = 2.8 Hz, 1H), 7.77 (dd, J = 8.8, 2.8 Hz, 1H), 7.68 - 7.59 (m, 2H), 7.28 (d, J = 8.8 Hz, 1H), 7.77 (dd, J = 7.6, 1.7 Hz, 1H), 6.75 (dd, J = 7.6, 1.7 Hz, 1H), 4.94 (s, 2H), 2.90 (t, J = 4.8 Hz, 4H), 2.54 (s, 4H), 2.25 (s, 3H). HRMS (ESI): calcd. for m / z C 15 H 20 N6O2, [M+H] +329.1720, found 329.1722. HPLC (100% MeOH): t R = 4.151 min, 95.29 %.

[0082] Example 13

[0083]

[0084] 6-chloro-N-(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)-5-nitropyrimidin-4-amine

[0085] Following the preparation procedure of Example 1, Step 2, replacing 4-chloroquinazoline with 4,6-dichloro-5-nitropyrimidine to give a yellow solid in 53.2% yield; m.p. >250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 10.20 (s, 1H), 8.85 (d, J = 6.5 Hz, 1H), 8.16 (d, J = 6.0 Hz, 2H), 7.62 (t, J = 9.5 Hz, 1H), 3.52 (br s, 4H), 2.42 (br s, 4H), 2.22 (s, 3H). HRMS (ESI): calcd. for m / z C 15 H 16 ClN7O4, [M+H] + 394.0671, found 394.0674. HPLC (100% MeOH): t R = 6.083 min, 97.01 %.

[0086] Example 14

[0087]

[0088] 4-chloro-6-((2-(4-methylpiperazin-1-yl)-5-nitrophenyl)amino)pyrimidine-5- carbonitrile

[0089] Following the preparation procedure of Example 1, Step 2, replacing 4-chloroquinazoline with 4,6-dichloro-5-nitropyrimidine to give a yellow solid in 53.2% yield; m.p. >250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 10.20 (s, 1H), 8.85 (d, J = 6.5 Hz, 1H), 8.16 (d, J = 6.0 Hz, 2H), 7.62 (t, J = 9.5 Hz, 1H), 3.52 (br s, 4H), 2.42 (br s, 4H), 2.22 (s, 3H). HRMS (ESI): calcd. for m / z C 16 H16 CIN7O2, [M+H] + 374.1133, found 374.1135. HPLC (100% MeOH): t R = 5.679 min, 98.83%.

[0090] Example 15

[0091]

[0092] 4-Chloro-6-((2-(4-methylpiperazin-1-yl)-5-nitrophenyl)amino)pyrimidine-5- carbaldehyde

[0093] Following the preparation procedure of Example 1, Step 2, replacing 4-chloroquinazoline with 4,6-dichloropyrimidine-5-carboxaldehyde to give a yellow solid in 75.9% yield; m.p. >250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 11.93 (s, 1H), 10.05 (s, 1H), 9.05 (d, J = 2.6 Hz, 1H), 8.36 (d, J = 1.6 Hz, 1H), 8.01 (dt, J = 8.9, 2.3 Hz, 1H), 7.36 (dd, J = 8.9, 1.8 Hz, 1H), 3.04 - 2.87 (m, 4H), 2.56 (t, J = 4.6 Hz, 4H), 2.26 (s, 3H). HRMS (ESI): calcd. for m / z C 16 H 17 CIN6O3, [M+H] + 377.1130, found 377.1132. HPLC (100% MeOH): t R = 6.472 min, 98.17%.

[0094] Example 16

[0095]

[0096] 6-Chloro-5-methoxy-N-(2-(4-methylpiperazin-1-yl)-5-nitrophenyl)pyrimidin-4- amine

[0097] Following the preparation procedure of Example 1, Step 2, replacing 4-chloroquinazoline with 4,6-dichloropyrimidine-5-carboxaldehyde to give a yellow solid in 75.9% yield; m.p. >250 °C; 1H NMR (300 MHz, CDC13) δ 9.59 (d, J = 2.7 Hz, 1H), 8.68 (s, 1H), 8.45 (d, J = 3.5 Hz, 1H), 7.96 (dd, J = 8.8, 2.7 Hz, 1H), 7.26 (d, J = 2.1 Hz, 1H), 4.06 (s, 1H), 3.06 - 2.97 (m, 4H), 2.67 (br s, 4H), 2.42 (s, 3H). HRMS (ESI): calcd. for m / z C 16 H 19 ClN6O3, [M+H] + 379.1280, found 379.1286. HPLC (100% MeOH): t R = 6.614 min, 95.68%.

[0098] Example 17

[0099]

[0100] 6-chloro-N 4 -(5-nitro-2-(piperazin-l-yl)phenyl)pyrimidine-4,5-diamine

[0101] Step 1: Preparation of 6-chloro-N 4 -(2-fluoro-5-nitrophenyl)pyrimidine-4,5-diamine

[0102] Dissolve 2-fluoro-5-nitroaniline (2 g, 12.8 mmol) in isopropanol (50 mL), add 2 mL of concentrated sulfuric acid and 4,6-dichloro-5-aminopyrimidine (25.6 mmol), reflux for 18 h, spin the reaction liquid to have solid precipitate, cool the precipitated solid, suction filter, wash the filter cake with isopropanol (5 mL x 3), and dry to obtain a yellow solid. Yield 59.5%; m.p. 209-211 °C; 1 H-NMR (300 MHz, DMSO-d6) δ 9.14 (s, 1H), 8.65-8.62 (m, 1H), 8.06-8.03 (m, 1H), 7.86 (s, 1H), 7.58-7.54 (m, 1H), 5.54 (s, 2H). m / z (EI-MS): 284.1 [M] + .

[0103] Step 2: Preparation of the target product 6-chloro-N 4 -(5-nitro-2-(piperazin-l-yl)phenyl)pyrimidine-4,5-diamine

[0104] Dissolve 6-chloro-N 4(2-morpholino-5-nitrophenyl)pyrimidine-4,5-diamine (0.35 mmol) was dissolved in 5 mL DMF, DIPEA (1.5 mmol) and morpholine (1.0 mmol) were added, and the reaction was carried out at 50 °C for 5 h. The reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL x 3), and saturated sodium chloride. The organic layer was dried over anhydrous sodium sulfate, and rotary evaporation was performed. The obtained crude product was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (2 mL) was added dropwise. The reaction was carried out at room temperature for 2 h. The reaction solution was neutralized with saturated sodium bicarbonate until no gas bubbles were generated, and ethyl acetate was added. Rotary evaporation was performed to obtain a yellow solid, which was washed with CH3CN and dried. The yield was 71.4%; m.p. > 250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.09 (br s, 1H), 8.60 (d, J = 2.7 Hz, 1H), 8.33 (s, 1H), 7.89 (dd, J = 8.9, 2.7 Hz, 1H), 7.83 (s, 1H), 7.23 (d, J = 8.9 Hz, 1H), 5.55 (s, 2H), 3.21 (s, 4H), 3.42 (t, J = 6.0 Hz, 4H). HRMS (ESI): calcd. for m / z C 14 H 17 ClN7O2,[M+H] + 350.1127, found 350.1131.HPLC (100% MeOH): t R = 4.069 min, 96.17%.

[0105] Example 18

[0106]

[0107] 6-chloro-N 4 -(2-morpholino-5-nitrophenyl)pyrimidine-4,5-diamine

[0108] 6-chloro-N 4 -(2-morpholino-5-nitrophenyl)pyrimidine-4,5-diamine (0.35 mmol) was dissolved in 5 mL DMF, DIPEA (1.5 mmol) and morpholine (1.0 mmol) were added, and the reaction was carried out at 50 °C for 5 h. The reaction solution was diluted with ethyl acetate (20 mL), washed with water (10 mL x 3), and saturated sodium chloride. The organic layer was dried over anhydrous sodium sulfate, and rotary evaporation was performed. The obtained crude product was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (2 mL) was added dropwise. The reaction was carried out at room temperature for 2 h. The reaction solution was neutralized with saturated sodium bicarbonate until no gas bubbles were generated, and ethyl acetate was added. Rotary evaporation was performed to obtain a yellow solid, which was washed with CH3CN and dried. The yield was 71.4%; m.p. > 250 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.70 (s, 1H), 8.32 (s, 1H), 8.08 - 7.86 (m, 2H), 7.29 (d, J = 9.0 Hz, 1H), 5.46 (s, 2H), 3.73 (s, 4H), 2.99 (s, 4H). HRMS (ESI): calcd. for m / z C 14 H 16 ClN6O3, [M+H] + 351.0967, found 351.0967. HPLC (100% MeOH): t R = 7.041 min, 98.84%.

[0109] Example 19

[0110]

[0111] 6-chloro-N 4 -(2-(4-ethylpiperazin-1-yl)-5-nitrophenyl)pyrimidine-4,5-diamine

[0112] Following the preparation procedure of Example 18, Step 2, using 1- ethylpiperazine in place of morpholine as a yellow solid, yield 46.7%; m.p. 185 °C - 187 °C; 1 H NMR (300 MHz, CDCl3) δ 9.34 (s, 1H), 8.32 (s, 1H), 8.18 (s, 1H), 7.93 (dt, J = 8.8, 2.7 Hz, 1H), 7.27 - 7.20 (m, 1H), 3.60 (s, 2H), 3.08 - 2.93 (m, 4H), 2.70 (s, 4H), 2.53 (qd, J = 7.2, 2.7 Hz, 2H), 1.16 (td, J = 7.3, 2.7 Hz, 3H). HRMS (ESI): calcd. for m / z C 16 H 21 ClN7O2, [M+H] + 378.1440, found 348.1449. HPLC (100% MeOH): t R = 4.542 min, 97.65%.

[0113] Example 20

[0114]

[0115] 6-chloro-N 4 -(2-(4-cyclopropylpiperazin-1-yl)-5-nitrophenyl)pyrimidine-4,5-diamine

[0116] Yellow solid, yield 58.9%; m.p. 223-225 °C; according to the procedure of Example 18, Step 2, replacing morpholine with 1-cyclopropylpiperazine. 1 H NMR (300 MHz, CDC13) δ 9.35 (d, J = 2.7 Hz, 1H), 8.32 (s, 1H), 8.21 (s, 1H), 7.92 (dd, J = 8.7, 2.7 Hz, 1H), 3.62 (s, 2H), 2.96 (t, J = 4.7 Hz, 4H), 2.87 (s, 4H), 1.78 - 1.71 (m, 1H), 0.56 - 0.51 (m, 2H), 0.49 - 0.44 (m, 2H). HRMS (ESI): calcd. for m / z C 17 H 21 ClN7O2, [M+H] + 390.1440, found 390.1446. HPLC (100% MeOH): t R = 4.811 min, 99.31%.

[0117] Example 21

[0118]

[0119] 2-(4-(2-((5-Amino-6-chloropyrimidin-4-yl)amino)-4-nitrophenyl)piperazin-1-yl)ethan-1-ol

[0120] Yellow solid, yield 76.4%; m.p. 202-204 °C; according to the procedure of Example 18, Step 2, replacing morpholine with 2-(piperazin-1-yl)ethan-1-ol. 1 H NMR (300 MHz, CDC13) δ 9.33 (s, 1H), 8.32 (s, 1H), 8.15 (s, 1H), 7.94 (d, J = 8.7 Hz, 1H), 7.27 (s, 1H), 3.70 (s, 2H), 3.64 - 2.46 (m, 2H), 3.02 (s, 4H), 2.78 (s, 4H), 2.68 (s, 2H), 2.57 (s, 1H). HRMS (ESI): calcd. for m / z C 16 H 20 ClN7O3, [M+H] + 394.1389, found 394.1387. HPLC (100% MeOH): t R = 7.347 min, 96.12%.

[0121] Example 22

[0122]

[0123] 6-chloro-N 4 -(5-nitro-2-(4-phenylpiperazin-1-yl)phenyl)pyrimidine-4,5-diamine

[0124] Following the preparation procedure of Example 18, Step 2, using 1-phenylpiperazine in place of morpholine, yellow solid, yield 76.4%; m.p. >250 °C; 1 H NMR (300 MHz, CDC13) δ 9.39 (d, J = 2.7 Hz, 1H), 8.35 (s, 1H), 8.25 (s, 1H), 7.97 (dd, J = 8.9, 2.6 Hz, 1H), 7.39 - 7.28 (m, 3H), 7.06 - 6.90 (m, 3H), 3.54 (s, 2H), 3.44 (t, J = 4.9 Hz, 4H), 3.16 (t, J = 4.9 Hz, 4H). HRMS (ESI): calcd. for m / z C 20 H 20 ClN7O2, [M+H] + 426.1440, found 426.1449. HPLC (100% MeOH): t R = 4.905 min, 95.52%.

[0125] Example 23

[0126]

[0127] (3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)(morpholino)methanone

[0128] Step 1: Preparation of methyl 4'-fluoro-3'-nitro-[1,1'-biphenyl]-4-carboxylate

[0129] Methyl 4-bromo-1-fluoro-2-nitrobenzene (5.0 g, 22.8 mmol) was dissolved in 1,4-dioxane (100 mL), (4-(methoxycarbonyl)phenyl)boronic acid (27.3 mmol) was added, cesium carbonate (14.8 g, 45.6 mmol) and a catalytic amount of Pd(PPh3)2Cl2 were added, and refluxed under N2protection for 20 h. The catalyst and cesium carbonate were removed by filtration, and the crude product after drying was column chromatographed (dichloromethane:methanol = 50:1) to obtain a white solid, yield 69.9%; m.p. 145 °C-147 °C. 1H NMR (300 MHz, CDC13) δ 8.31-8.13 (m, 3H), 7.88-7.59 (m, 1H), 8.28 (m, 3H), 7.42-7.36 (m, 1H), 3.94 (s, 3H). m / z (EI-MS): 276.2 [M] + .

[0130] Step 2: Preparation of 4'-(4-methylpiperazin-1-yl)-3'-nitro-[1,1'-biphenyl]-4- carboxylic acid methyl ester

[0131] Following the preparation procedure of Example 1, Step 1, replace 2-F-5-nitroaniline with 4'-fluoro-3'-nitro-[1,1'-biphenyl]-4-carboxylic acid methyl ester to give a yellow solid in 93.8% yield; m.p. 130-132 °C; 1 H NMR (300 MHz, CDC13) δ 8.31-8.13 (m, 3H), 7.88-7.59 (m, 1H), 8.28 (m, 3H), 7.42-7.36 (m, 1H), 3.94 (s, 3H). m / z (EI-MS): 276.2 [M] + .

[0132] Step 3: Preparation of 3'-amino-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4- carboxylic acid methyl ester

[0133] Dissolve 4'-(4-methylpiperazin-1-yl)-3'-nitro-[1,1'-biphenyl]-4-carboxylic acid methyl ester (12.9 mmol) in methanol, add a catalytic amount of Pd / C, stir under H2protection at room temperature for 2 h, filter and dry to give an off-white solid. Yield 90.4%; m.p. 229-231 °C; 1 H NMR (300 MHz, CDC13) δ 8.31-8.13 (m, 3H), 7.88-7.59 (m, 1H), 8.28 (m, 3H), 7.42-7.36 (m, 1H), 3.94 (s, 3H). m / z (EI-MS): 276.2 [M] + .

[0134] Step 4: Preparation of methyl 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylate

[0135] Methyl 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-1- yl)-[1,1'-biphenyl]-4-carboxylate was prepared according to the procedure described in Example 1, Step 4, using methyl 3'-amino-4'-(4-methylpiperazin-1-yl)-[1,1'- biphenyl]-4-carboxylate (0.1 g, 0.28 mmol) and 4,6-dichloro-5-nitropyrimidine (0.1 g, 0.28 mmol) as starting materials. The crude product was purified by column chromatography on silica gel (dichloromethane:methanol = 100:1) to give a yellow solid. The yellow solid was dissolved in ethyl acetate (20 mL), and SnCl2(0.3 g, 2.8 mmol) was added. The mixture was refluxed for 4 h. The reaction mixture was cooled to room temperature, and saturated NaHC03solution was added until no more gas was generated. The mixture was filtered, and the filter cake was washed with ethyl acetate (4 x 20 mL). The filtrate was extracted with ethyl acetate (4 x 50 mL), and the combined organic layers were dried and concentrated to give a white solid. The yield was 45.7%; m.p. 209-210 °C. 1 H NMR (300 MHz, DMSO-d6) δ 8.53-8.42 (m, 1H), 8.23 (s, 1H), 7.96-7.90 (m, 3H), 7.70 (d, J = 8.4 Hz, 2H), 7.46-7.42 (m, 1H), 7.26 (d, J = 8.3 Hz, 1H), 5.36 (s, 2H), 3.93 (s, 3H), 3.57 (t, J = 4.6 Hz, 4H), 2.87 (t, J = 4.7 Hz, 4H), 2.24 (s, 3H). m / z (EI-MS): 453.2 [M] + .

[0136] Step 5: Preparation of 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid

[0137] Methyl 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-1- yl)-[1,1'-biphenyl]-4-carboxylate was prepared according to the procedure described in Example 1, Step 4, using methyl 3'-amino-4'-(4-methylpiperazin-1-yl)-[1,1'- biphenyl]-4-carboxylate (0.1 g, 0.28 mmol) and 4,6-dichloro-5-nitropyrimidine (0.1 g, 0.28 mmol) as starting materials. The crude product was purified by column chromatography on silica gel (dichloromethane:methanol = 100:1) to give a yellow solid. The yellow solid was dissolved in ethyl acetate (20 mL), and SnCl2(0.3 g, 2.8 mmol) was added. The mixture was refluxed for 4 h. The reaction mixture was cooled to room temperature, and saturated NaHC03solution was added until no more gas was generated. The mixture was filtered, and the filter cake was washed with ethyl acetate (4 x 20 mL). The filtrate was extracted with ethyl acetate (4 x 50 mL), and the combined organic layers were dried and concentrated to give a white solid. The yield was 45.7%; m.p. 209-210 °C. 1H NMR (300 MHz, DMSO-d6) δ 8.33-8.28 (m, 2H), 8.10 (s, 1H), 7.86-7.84 (m, 2H), 7.80 (s, 1H), 7.13-7.11 (m, 2H), 6.88 (d, J = 2.0 Hz, 1H), 5.42 (s, 2H), 3.54 (t, J = 4.6 Hz, 4H), 2.35 (t, J = 4.7 Hz, 4H), 2.24 (s, 3H). m / z (EI-MS): 439.3 [M] +

[0138] Step 6: Preparation of the target product: (3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)(morpholino)methanone

[0139] Methyl 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylate (0.1 g, 0.26 mmol) was dissolved in DMF (10 mL), and condensing agent BOP (0.22 g, 0.49 mmol) and triethylamine (0.12 mL, 0.88 mmol) were added. After stirring at room temperature for 30 min, morpholine (0.39 mmol) was added, and the reaction was stirred at room temperature for 12 h. The reaction solution was poured into water (50 mL), extracted with ethyl acetate (3 x 50 mL), and the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. Purification by column chromatography (dichloromethane:methanol = 50:1) gave a white solid in a yield of 67.8%; m.p. 225-227 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.45 (d, J = 13.6 Hz, 2H), 7.90 (s, 1H), 7.69 (d, J = 7.9 Hz, 2H), 7.49 (d, J = 7.7 Hz, 2H), 7.42 (d, J = 8.4 Hz, 1H), 7.27 (d, J = 8.2 Hz, 1H), 5.39 (s, 2H), 3.61 (s, 4H), 3.35 (s, 4H), 2.98-2.90 (s, 4H), 2.64 (s, 4H), 2.34 (s, 3H). HRMS (ESI): calcd. for m / z C 26 H 30 ClN7O2, [M+H] + 508.2222, found 508.2223. HPLC (80% methanol in water): t R = 3.507 min, 98.81%.

[0140] Example 24

[0141]

[0142] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-N-(2- morpholinoethyl)-[1,1'-biphenyl]-4-carboxamide

[0143] According to the preparation method of Example 23, Step 6, using 2-morpholinoethyl-1- amine instead of morpholine, a white solid was obtained in 55.8% yield; m.p. 230-232 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.27-8.22 (m, 2H), 7.92-7.90 (m, 3H), 7.71 (d, J = 8.0 Hz, 2H), 7.45 (d, J = 8.2 Hz, 1H), 7.26 (d, J = 8.4 Hz, 1H), 5.39 (s, 2H), 3.58 (s, 4H), 3.40-3.34 (m, 4H), 2.90 (s, 4H), 2.61 (s, 4H), 2.44 (s, 4H), 2.31 (s, 3H). HRMS (ESI): calcd. for m / z C 28 H 35 ClN8O2, [M+H] + 551.2644, found 551.2639. HPLC (80% methanol in water): t R = 3.196 min, 98.91%.

[0144] Example 25

[0145]

[0146] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-N-(3- morpholinopropyl)-[1,1'-biphenyl]-4-carboxamide

[0147] According to the preparation method of Example 23, Step 6, using 3-morpholinopropyl-1- amine instead of morpholine, a white solid was obtained in 48.9% yield; m.p. 210-212 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.53 - 8.42 (m, 1H), 8.28 (d, J = 2.8 Hz, 1H), 8.23 (s, 1H), 7.96 - 7.90 (m, 3H), 7.70 (d, J = 8.4 Hz, 2H), 7.46 - 7.42 (m, 1H), 7.26 (d, J = 8.3 Hz, 1H), 5.36 (s, 2H), 3.57 (t, J = 4.6 Hz, 4H), 3.32 - 3.30 (m, 2H), 2.87 (t, J = 4.7 Hz, 4H), 2.51 - 2.49 (m, 4H), 2.37 - 2.32 (m, 6H), 2.24 (s, 3H). HRMS (ESI): calcd. for m / z C 29 H 31 ClN8O2, [M+H] + 565.2801, found 565.2801. HPLC (80% methanol in water): t R = 3.974 min, 96.00%.

[0148] Example 26

[0149]

[0150] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-N,N-dimethyl-4"-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxamide

[0151] White solid, yield 78.9% according to the preparation method of step 6 of example 23, replacing morpholine with dimethylamine; m.p. 142-145 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.53 - 8.42 (m, 1H), 8.28 (d, J = 2.8 Hz, 1H), 8.23 (s, 1H), 7.96 - 7.90 (m, 3H), 7.70 (d, J = 8.4 Hz, 2H), 7.46 - 7.42 (m, 1H), 7.26 (d, J = 8.3 Hz, 1H), 5.36 (s, 2H), 3.57 (t, J = 4.6 Hz, 4H), 3.32 - 3.30 (m, 2H), 2.87 (t, J = 4.7 Hz, 4H), 2.51 - 2.49 (m, 4H), 2.37 - 2.32 (m, 6H), 2.24 (s, 3H). HRMS (ESI): calcd. for m / z C 24 H 28 ClN7O, [M+H] + 466.2117, found 466.2121. HPLC (80% methanol in water): t R = 2.489 min, 95.56%.

[0152] Example 27

[0153]

[0154] (3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)(4-methylpiperazin-1-yl)methanone

[0155] According to the preparation method of Example 23, Step 6, using 1-methylpiperazine instead of morpholine to give a white solid in 56.9% yield; m.p. >250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.54 (d, J = 13.6 Hz, 2H), 7.90 (s, 1H), 7.68 (d, J = 7.9 Hz, 2H), 7.45 (d, J = 7.7 Hz, 2H), 7.39 (d, J = 8.4 Hz, 1H), 7.27 (d, J = 8.1 Hz, 1H), 5.35 (s, 2H), 3.61 (s, 4H), 3.35 (s, 4H), 2.98-2.90 (s, 4H), 2.64 (s, 4H), 2.34 (s, 3H), 2.30 (s, 3H). HRMS (ESI): calcd. for m / z C 27 H 33 ClN8O, [M+H] + 521.2937, found 521.2937. HPLC (80% methanol in water): t R = 4.094 min, 98.10%.

[0156] Example 28

[0157]

[0158] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-N-(3-(dimethylamino)propyl)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxamide

[0159] According to the preparation method of Example 23, Step 6, using N 1 , N 1 -dimethyl-1,3-diamine instead of morpholine to give a white solid in 38.9% yield; m.p. 232 °C-234 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.66 (s, 1 H), 8.26-8.20 (m, 2 H), 7.93-7.88 (m, 3 H), 7.73 (d, J = 9.8 Hz, 2 H), 7.47 (s, 1 H), 7.29 (s, 1 H), 5.44 (s, 2 H), 3.08-2.90 (m, 8 H), 2.77 (s, 6 H), 2.50 (s, 4 H), 1.90 (s, 4 H), 1.28-1.23 (m, 2 H). HRMS (ESI): calcd. for m / z C 27 H 36 ClN8O,[M+H] + 523.2695, found 523.2695.HPLC (90% methanol in water): t R = 3.409 min, 98.88%.

[0160] Example 29

[0161]

[0162] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-N-(2-(pyrrolidin-1-yl)ethyl)-[1,1 '-biphenyl]-4-carboxamide

[0163] Following the preparation procedure of Example 23, Step 6, using 2-(pyrrolidin-1-yl)ethan-1-amine in place of morpholine to give a white solid in 37.8% yield; m.p. 241 °C-243 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.66 (s, 1 H), 8.26-8.20 (m, 2 H), 7.93-7.88 (m, 3 H), 7.73 (d, J = 9.8 Hz, 2 H), 7.47 (s, 1 H), 7.29 (s, 1 H), 5.44 (s, 2 H), 3.08-2.90 (m, 8 H), 2.77 (s, 6 H), 2.50 (s, 4 H), 1.90 (s, 4 H), 1.28-1.23 (m, 2 H). HRMS (ESI): calcd. for m / z C 28 H 35 ClN8O,[M+H] + 535.2695, found 535.2690.HPLC (100% methanol): t R = 7.516 min, 98.64%.

[0164] Example 30

[0165]

[0166] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-N-(3-hydroxypropyl)-4"-(4- methylpiperazin- 1 -yl)- [ 1, 1 '-biphenyl] -4-carboxamide

[0167] Following the procedure of Example 23, Step 6, using 3-aminopropan-1-ol in place of morpholine as a white solid in 50.7% yield; m.p. 189-192 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.45 (d, J = 7.2 Hz, 1H), 8.30 - 8.19 (m, 2H), 7.93 - 7.90 (m, 3H), 7.72 - 7.69 (m, 2H), 7.43 (d, J = 8.4 Hz, 1H), 7.26 (d, J = 8.4 Hz, 1H), 5.37 (s, 2H), 4.49 - 4.47 (m, 1H), 3.51 - 3.45 (m, 4H), 2.88 (s, 4H), 2.52 - 2.50 (m, 4H), 2.27 (s, 3H), 1.74 - 1.64 (m, 2H). HRMS (ESI): calcd. for m / z C 25 H 31 ClN7O2, [M+H] + 496.2222, found 496.2217. HPLC (80% methanol in water): t R = 2.384 min, 96.63%

[0168] Example 31

[0169]

[0170] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-N-iso-propyl-4"-(4-methylpiperazin- 1 -yl)- [ 1, 1 '-biphenyl] -4-carboxamide

[0171] Following the procedure of Example 23, Step 6, using 3-aminopropan-1-ol in place of morpholine as a white solid in 50.7% yield; m.p. 189-192 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.27 - 8.22 (m, 2H), 7.91 (d, J = 8.1 Hz, 2H), 7.71 (s, 1H), 7.59 (d, J = 8.0 Hz, 2H), 7.43 - 7.30 (m, 2H), 7.04 (d, J = 8.2 Hz, 1H), 5.54 (s, 2H), 4.12 (s, 1H), 3.33 (s, 4H), 3.07 (s, 4H), 2.82 (s, 3H), 1.18 (s, 6H). HRMS (ESI): calcd. for m / z C 25 H 30 ClN7O,[M+H] + 480.2279, found 480.2276.HPLC (80% methanol in water): t R = 3.601 min, 96.77%.

[0172] Example 32

[0173]

[0174] Step 1: Preparation of 1-(4-bromo-2-nitrophenyl)-4-methylpiperazine

[0175] 2-Fluoro-5-bromonitrobenzene (10.0 g, 45.4 mmol) was dissolved in DMF (20 mL), DIPEA (9.5 mL, 54.6 mmol) and 1-methylpiperazine (6.6 mL, 54.6 mmol) were added, after stirring at 80 °C for 8 h, the reaction was poured into 100 mL water, extracted with ethyl acetate (3 x 100 mL), the organic layer was dried with anhydrous sodium sulfate, filtered, and rotary evaporated to give a yellow solid, yield 95.0%; 1 H NMR (300 MHz, DMSO-d6) δ 8.27 - 8.22 (m, 2H), 7.91 (d, J = 8.1 Hz, 2H), 7.71 (s, 1H), 7.59 (d, J = 8.0 Hz, 2H), 7.43 - 7.30 (m, 2H), 7.04 (d, J = 8.2 Hz, 1H), 5.54 (s, 2H), 4.12 (s, 1H), 3.33 (s, 4H), 3.07 (s, 4H), 2.82 (s, 3H), 1.18 (s, 6H). HRMS (ESI): calcd. for m / z C + .

[0176] Step 2: Preparation of 5-bromo-2-(4-methylpiperazin-1-yl)aniline

[0177] To a solution of 1-(4-bromo-2-nitrophenyl)-4-methylpiperazine (12.9 g, 43.14 mmol) in methanol (100 mL) was added a catalytic amount of Pd / C under hydrogen atmosphere. The reaction mixture was stirred at room temperature for 2 h. The mixture was filtered and the filtrate was concentrated to give a yellow solid in 89% yield; m.p. 165-167 °C; 1 H NMR (300 MHz, CDC13) δ 6.84-6.79 (m, 3H), 4.01 (s, 2H), 2.89 (s, 4H), 2.45 (s, 4H), 2.35 (s, 3H). m / z (EI-MS): 270.0 [M] + .

[0178] Step 3: Preparation of N 4 -(5-bromo-2-(4-methylpiperazin-1-yl)phenyl)-6-chloropyrimidine-4,5-diamine

[0179] According to the preparation method of Step 4 of Example 23, 3'-amino-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester was replaced by 5- bromo-2-(4-methylpiperazin-1-yl)aniline to give a white solid. Yield 72.5%; m.p. 196- 198 °C; 1 H NMR (300 MHz, CDC13) δ 6.84-6.79 (m, 3H), 4.01 (s, 2H), 2.89 (s, 4H), 2.45 (s, 4H), 2.35 (s, 3H). m / z (EI-MS): 270.0 [M] + .

[0180] Step 4: Preparation of the target product N 4 -(4'-amino-4-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-3-yl)-6-chloropyrimidine-4,5- diamine

[0181] To a solution of N 4 -(5-bromo-2-(4-methylpiperazin-1-yl)phenyl)-6-chloropyrimidine-4,5-diamine (1.0 g, 2.5 mmol) in 1,4-dioxane (30 mL) was added 4-aminobenzoic acid (2.9 mmol) and cesium carbonate (1.6 g, 5 mmol) and a catalytic amount of Pd(PPh3)2Cl2 under nitrogen atmosphere. The reaction mixture was refluxed for 20 h. The catalyst and cesium carbonate were removed by filtration. The filtrate was concentrated and purified by column chromatography (dichloromethane:methanol = 50:1) to give a yellow solid in 67.3% yield; m.p. 212 °C-214 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.16 (d, J = 8.1 Hz, 2H), 7.90 (s, 1H), 7.65 (d, J = 8.2 Hz, 2H), 7.55 (d, J = 8.4 Hz, 2H), 7.37 (d, J = 8.4 Hz, 1H), 7.26 (d, J = 8.2 Hz, 1H), 5.43 (s, 2H), 5.15 (s, 2H), 3.17 (s, 4H), 3.06 (s, 4H), 2.75 (s, 3H). m / z (EI-MS): 410.2 [M] + HPLC (100% methanol): t R = 7.489 min, 95.67%.

[0182] Example 33

[0183]

[0184] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)tetrahydro-2H-pyran-4-carboxamide

[0185] Tetrahydro-2H-pyran-4-carboxylic acid (0.35 mmol) was dissolved in DMF (8 mL), BOP (0.26 g, 0.59 mmol) and triethylamine (0.1 mL, 0.88 mmol) were added, after stirring at room temperature for 30 min, Example 32 (0.12 g, 0.29 mmol) was added, stirring at room temperature for 12 h, the reaction solution was poured into 50 mL of water, extracted with ethyl acetate (3 x 50 mL), dried over anhydrous sodium sulfate, filtered, rotary evaporated, separated and purified by silica gel column chromatography (dichloromethane:methanol = 50:1) to give a white solid, yield 78.5%; m.p. 207-209 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.99 (s, 1H), 8.21 (s, 1H), 7.90 (s, 1H), 7.68 (dd, J = 8.4, 5.9 Hz, 3H), 7.57-7.54 (m, 2H), 7.38-7.35 (m, 1H), 5.38 (s, 2H), 3.94-3.90 (m, 3H), 3.37-3.35 (m, 2H), 2.92 (s, 4H), 2.76 (s, 4H), 2.43 (s, 3H), 1.72-1.69 (m, 4H). HRMS (ESI): calcd. for m / z C 27 H 32 ClN7O2, [M+H] +522.2379, found 522.2377. HPLC (80% methanol in water): t R = 4.280 min, 95.50%.

[0186] Example 34

[0187]

[0188] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)-1-methylpiperidine-4-carboxamide

[0189] According to Example 33, replace tetrahydro-2H-pyran-4-carboxylic acid with 1-methylpiperidine-4-carboxylic acid to give a white solid in 63.6% yield; m.p. 157-160 °C; 1 H NMR (300 MHz, DMSO-d6) δ 10.19 (s, 1H), 8.22-8.19 (m, 2H), 7.95 (s, 1H), 7.69 (d, J = 8.4 Hz, 2H), 7.57 (d, J = 8.7 Hz, 2H), 7.36 (d, J = 8.4 Hz, 1H), 7.24 (d, J = 8.1 Hz, 1H), 5.42 (s, 2H), 2.99-2.90 (m, 5H), 2.85 (s, 4H), 2.73 (s, 3H), 2.53 (s, 4H), 2.49 (s, 3H), 2.03-1.99 (m, 4H). HRMS (ESI): calcd. for m / z C 28 H 36 ClN8O, [M+H] + 535.2695, found 535.2710. HPLC (80% methanol in water): t R = 4.518 min, 98.25%.

[0190] Example 35

[0191]

[0192] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)cyclohexanecarboxamide

[0193] According to Example 33, replace tetrahydro-2H-pyran-4-carboxylic acid with cyclohexanecarboxylic acid to give a white solid in 80.1% yield; m.p. 256-258 °C; 1H NMR (300 MHz, DMSO-d6) δ 9.91 (s, 1H), 8.18 (d, J = 8.2 Hz, 2H), 7.90 (s, 1H), 7.69 (d, J = 8.3 Hz, 2H), 7.55 (d, J = 8.4 Hz, 2H), 7.37 (d, J = 8.4 Hz, 1H), 7.26 (d, J = 8.2 Hz, 1H), 5.43 (s, 2H), 3.17 (s, 4H), 3.06 (s, 4H), 2.75 (s, 3H), 2.34 - 2.30 (m, 1H), 1.76 (t, J = 12.5 Hz, 4H), 1.42 (t, J = 11.6 Hz, 2H), 1.33 - 1.09 (m, 4H). HRMS (ESI): calcd. for m / z C 28 H 34 ClN7O,[M+H] + 520.2586, found 520.2584. HPLC (90% methanol in water): t R = 4.003 min, 97.87%.

[0194] Example 36

[0195]

[0196] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)butanamide

[0197] According to Example 33, replacing tetrahydro-2H-pyran-4-carboxylic acid with n-butyric acid, white solid was obtained in 79.7% yield; m.p. 196 °C-198 °C; 1 H NMR (300 MHz, DMSO-d6) δ 10.00 (s, 1H), 8.21 - 8.16 (m, 2H), 7.90 (d, J = 6.0 Hz, 1H), 7.71 - 7.69 (m, 2H), 7.59 - 7.54 (m, 2H), 7.38 (d, J = 7.8 Hz, 1H), 7.27 (d, J = 8.2 Hz, 1H), 5.44 (s, 2H), 3.51 - 3.46 (m, 4H), 3.27 - 3.16 (m, 4H), 2.88 (s, 3H), 2.32 - 2.27 (m, 2H), 1.68 - 1.58 (m, 2H), 0.96 - 0.89 (m, 3H). HRMS (ESI): calcd. for m / z C 25 H 31 ClN7O,[M+H] +480.2273, found 480.2274. HPLC (80% methanol in water): t R = 4.696 min, 96.97%.

[0198] Example 37

[0199]

[0200] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)- [1,1'-biphenyl]-4-yl)benzamide

[0201] According to Example 33, tetrahydro-2H-pyran-4-carboxylic acid was replaced by benzoic acid to give a white solid in 77.9% yield; m.p. > 250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 10.00 (s, 1H), 8.21 - 8.16 (m, 2H), 7.90 (d, J = 6.0 Hz, 1H), 7.71 - 7.69 (m, 2H), 7.59 - 7.54 (m, 2H), 7.38 (d, J = 7.8 Hz, 1H), 7.27 (d, J = 8.2 Hz, 1H), 5.44 (s, 2H), 3.51 - 3.46 (m, 4H), 3.27 - 3.16 (m, 4H), 2.88 (s, 3H), 2.32 - 2.27 (m, 2H), 1.68 - 1.58 (m, 2H), 0.96 - 0.89 (m, 3H). HRMS (ESI): calcd. for m / z C 28 H 28 ClN7O, [M+H] + 514.2117, found 514.2115. HPLC (90% methanol in water): t R = 3.889 min, 95.09%.

[0202] Example 38

[0203]

[0204] (3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2-fluoro-4"-(4-methylpiperazin-1-yl)- [1,1'-biphenyl]-4-yl)(4-methylpiperazin-1-yl)methanone

[0205] Following the procedure of Example 32, Step 4, substituting 4-aminobenzoic acid for 2-F-4-carboxybenzoic acid, the white intermediate was prepared. Following the procedure of Example 23, Step 6, substituting morpholine for 1-methylpiperazine, the white solid was prepared in 43.1% yield; m.p. 182-184 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.12 (s, 1H), 7.80 (s, 1H), 7.68-7.65 (m, 2H), 7.60 (dd, J = 7.5, 2.0 Hz, 1H), 7.10 (dd, J = 7.5, 2.0 Hz, 1H), 6.93 (d, J = 7.3 Hz, 1H), 6.82 (d, J = 2.0 Hz, 1H), 5.36 (s, 2H), 3.55 (t, J = 5.1 Hz, 4H), 3.30 (t, J = 5.1 Hz, 4H), 2.85 (t, J = 4.7 Hz, 4H), 2.53 (s, 4H), 2.34 (s, 3H), 2.32 (s, 3H). HRMS (ESI): calcd. for m / z C 27 H 32 ClFN8O,[M+H] + 539.2444, found 539.2443.HPLC (80% methanol in water): t R = 4.156 min, 95.21%.

[0206] Example 39

[0207]

[0208] N 4 -(3'-Amino-4-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-3-yl)-6-chloropyrimidine electronic-4,5-diamine

[0209] Following the procedure of Example 32, Step 4, substituting 4-aminobenzoic acid for 2-F-4-carboxybenzoic acid, the white intermediate was prepared. Following the procedure of Example 23, Step 6, substituting morpholine for 1-methylpiperazine, the white solid was prepared in 43.1% yield; m.p. 182-184 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.23 (s, 1H), 8.17 (s, 1H), 7.90 (s, 1H), 7.23 (d, J = 6.4 Hz, 2H), 7.07 (d, J = 7.7 Hz, 1H), 6.80 (s, 1H), 6.73 (d, J = 7.6 Hz, 1H), 6.53 (d, J = 8.0 Hz, 1H), 5.36 (s, 2H), 5.17 (s, 2H), 2.85 (s, 4H), 2.61 (s, 4H), 2.25 (s, 3H). HRMS (ESI): calcd. for m / z C 21 H 24 ClN7,[M+H] + 410.1854, found 410.1852.HPLC (100% methanol): t R = 7.491 min, 95.66%.

[0210] Example 40

[0211]

[0212] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-3-yl)tetrahydro-2H-pyran-4-carboxamide

[0213] According to the preparation method of Example 33, replace Example 32 with Example 39 to obtain a white solid with a yield of 65.0%; m.p. 212 °C-214 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.99 (s, 1H), 8.24 (s, 1H), 8.19 (s, 1H), 7.89 (s, 2H), 7.59 (d, J = 8.0 Hz, 1H), 7.44-7.19 (m, 4H), 5.38 (s, 2H), 3.91 (d, J = 10.9 Hz, 2H), 3.47-3.35 (m, 3H), 2.86 (s, 4H), 2.54 (m, 4H), 2.24 (s, 3H), 1.70 (s, 4H). HRMS (ESI): calcd. for m / z C 27 H 32 ClN7O2,[M+H] + 522.2379, found 522.2368.HPLC (100% methanol): t R = 7.105 min, 98.72%.

[0214] Example 41

[0215]

[0216] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-3-yl)-1-methylpiperidine-4-carboxamide

[0217] Following the preparation procedure of Example 34, replace Example 32 with Example 39 to give a white solid in 56.9% yield; m.p. 158-160 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.95 (s, 1H), 8.24 (s, 1H), 8.18 (s, 1H), 7.89 (s, 2H), 7.59 (d, J = 8.3 Hz, 1H), 7.39 - 7.19 (m, 4H), 5.38 (s, 2H), 2.86 - 2.83 (m, 4H), 2.80 (s, 1H), 2.57 - 2.51 (m, 4H), 2.22 (s, 3H), 2.15 (s, 3H), 1.89 - 1.82 (m, 2H), 1.79 - 1.58 (m, 4H), 1.22 (s, 2H). HRMS (ESI): calcd. for m / z C 28 H 36 ClN8O, [M+H] + 535.2695, found 535.2702. HPLC (100% methanol): t R = 7.252 min, 95.45%.

[0218] Example 42

[0219]

[0220] 6-chloro-N 4 -(4'-methoxy-4-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-3-yl)pyrimidine-4,5-diamine

[0221] Following the preparation procedure of Example 32, step 4, replace 4- aminobenzoic acid with 4-methoxybenzoic acid to give a white solid in 59.9% yield; m.p. 211-213 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.24 (s, 2H), 7.90 (s, 1H), 7.57 (d, J = 7.9 Hz, 2H), 7.40-7.35 (m, 3H), 7.24 (d, J = 8.3 Hz, 1H), 5.37 (s, 2H), 5.22 (s, 1H), 4.52 (s, 2H), 2.85 (t, J = 4.5 Hz, 4H), 2.53 (s, 4H), 2.24 (s, 3H). m / z (EI-MS): 425.2 [M] + HPLC (90% methanol in water): tR= 6.859 min, 96.26%. R

[0222] Example 43

[0223]

[0224] 6-chloro-N 4 -(5-(furan-3-yl)-2-(4-methylpiperazin-1-yl)phenyl)pyrimidine-4,5-diamine

[0225] Following the preparation procedure of Example 32, Step 4, replace 4- aminobenzoic acid with 3-furanbenzoic acid to give a white solid in 70.1% yield; m.p. 164-166 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.23 (s, 1H), 7.90 (s, 1H), 7.54 (d, J = 7.5 Hz, 1H), 7.43 (s, 1H), 7.07 (d, J = 7.7 Hz, 1H), 6.91 (d, J = 7.4 Hz, 1H), 6.82 (s, 1H), 6.53 (d, J = 8.0 Hz, 1H), 5.36 (s, 2H), 2.85 (s, 4H), 2.61 (s, 4H), 2.25 (s, 3H). HRMS (ESI): calcd. for m / z C 19 H 22 ClN6O, [M+H] + 385.1538, found 385.1538. HPLC (90% methanol in water): tR= 6.859 min, 96.26%. R

[0226] Example 44

[0227]

[0228] 6-chloro-N 4 ​​6-chloro-N

[0229] Following the preparation procedure of Example 32, Step 4, replace 4- aminobenzoic acid with 4-pyridinebenzoic acid to give a white solid in 44.2% yield; m.p. 202-204 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.17 (s, 1H), 9.10 (s, 2H), 8.57 (s, 1H), 7.80 (s, 1H), 7.10 (dd, J = 7.5, 2.0 Hz, 1H), 6.99 (d, J = 7.4 Hz, 1H), 6.82 (d, J = 2.0 Hz, 1H), 5.32 (s, 2H), 2.85 (s, 4H), 2.54 (s, 4H), 2.34 (s, 3H). HRMS (ESI): calcd. for m / z C 19 H 22 ClN8,[M+H] + 397.1651, found 397.1653.HPLC (80% methanol in water): t R = 3.973 min, 97.50%.

[0230] Example 45

[0231]

[0232] 6-chloro-N 4 -(2-(4-methylpiperazin-1-yl)-5-(pyridin-4-yl)phenyl)pyrimidine-4,5-diamine

[0233] Following the preparation procedure of Example 32, Step 4, replace 4- aminobenzoic acid with 4-pyridinebenzoic acid to give a white solid in 44.2% yield; m.p. 202-204 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.17 (s, 1H), 9.10 (s, 2H), 8.57 (s, 1H), 7.80 (s, 1H), 7.10 (dd, J = 7.5, 2.0 Hz, 1H), 6.99 (d, J = 7.4 Hz, 1H), 6.82 (d, J = 2.0 Hz, 1H), 5.32 (s, 2H), 2.85 (s, 4H), 2.54 (s, 4H), 2.34 (s, 3H). HRMS (ESI): calcd. for m / z C + .HPLC (100% methanol): tR = 7.074 min, 96.00 %.

[0234] Example 46

[0235]

[0236] 1 -(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin- 1 -yl)phenyl)- 1 H- 1,2,3-triazole-4-carboxylic acid methyl ester Step 1 : Preparation of 4-(4-methylpiperazin-1-yl)-3-nitroaniline

[0237] 4-F-3-nitroaniline (5 g, 32 mmol) was dissolved in acetonitrile (50 mL), DIPEA (7.9 mL, 47.8 mmol) and 1 -methylpiperazine (5.3 mL, 67.8 mmol) were added, refluxed for 12 h, the reaction was rotary evaporated to get red solid crude, washed with acetonitrile, suction filtered, oven dried, yield 95.5 %; m.p. > 250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 7.06 (d, J = 8.6 Hz, 1H), 6.76 (s, 1H), 6.69 (d, J = 8.5 Hz, 1H), 5.34 (s, 2H), 2.70 (t, J = 4.4 Hz, 4H), 2.27 (s, 4H), 2.09 (s, 3H). m / z (EI-MS): 237.1 [M] + .

[0238] Step 2: Preparation of 1 -(4-azido-2-nitrophenyl)-4-methylpiperazine

[0239] 4-(4-methylpiperazin-1-yl)-3-nitroaniline (5.0 g, 21.25 mmol) was dissolved in 2 M HCI, aqueous solution of NaNCte (2.2 g, 31.8 mmol) (12 mL) was added dropwise under ice bath condition, stirred for 30 min under ice bath condition, sodium azide (2.8 g, 42.5 mmol) was added, stirred for 2 h at room temperature, the reaction was adjusted to pH = 9-10 with 2 M NaOH to precipitate red-brown solid, filtered and dried, yield 87.7 %; m.p. 88-91 °C; 1 H NMR (300 MHz, DMSO-d6) δ 7.06 (d, J = 8.6 Hz, 1H), 6.76 (s, 1H), 6.69 (d, J = 8.5 Hz, 1H), 5.34 (s, 2H), 2.70 (t, J = 4.4 Hz, 4H), 2.27 (s, 4H), 2.09 (s, 3H). m / z (EI-MS): 237.1 [M] + .

[0240] Step 3: Preparation of methyl l-(4-(4-methylpiperazin-l-yl)-3-nitrophenyl)-lH- 1,2,3-triazole-4-carboxylate

[0241] Dissolve l-(4-azido-2-nitrophenyl)-4-methylpiperazine (2.0 g, 7.6 mmol) in methanol (50 mL) and add methyl propiolate (1.8 g, 22.8 mmol) followed by Cul (0.14 g, 0.76 mmol) and DIPEA (1.2 mL, 7.6 mmol) and reflux for 48 h. Filter off Cul and spin dry the filtrate to get crude product which is washed with ethyl acetate to get red brown solid. Yield 61.7%; m.p. 159-161 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.45 (s, 1H), 8.36 (d, J = 2.7 Hz, 1H), 8.11 - 8.01 (m, 1H), 7.42 (d, J = 9.1 Hz, 1H), 3.80 (s, 3H), 2.99 (t, J = 5.4 Hz, 4H), 2.35 (t, J = 5.2 Hz, 4H), 2.13 (s, 3H). m / z (EI-MS): 369.2 [M+Na] + .

[0242] Step 4: Preparation of methyl l-(3-amino-4-(4-methylpiperazin-l-yl)phenyl)-lH- 1,2,3-triazole-4-carboxylate

[0243] Dissolve methyl l-(4-(4-methylpiperazin-l-yl)-3-nitrophenyl)-lH-l,2,3-triazole-4- carboxylate (1.7 g, 4.9 mmol) in methanol (50 mL) and add catalytic amount of Pd / C and stir at room temperature under hydrogen atmosphere for 6 h. Filter off Pd / C and spin dry the filtrate to get pink solid. Yield 85.3%; m.p. 194-197 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.28 (s, 1H), 7.28 (d, J = 1.9 Hz, 1H), 7.07 (d, J = 1.9 Hz, 2H), 5.15 (s, 2H), 3.90 (s, 3H), 2.87 (t, J = 4.5 Hz, 4H), 2.53 (br s, 4H), 2.26 (s, 3H). m / z (EI-MS): 317.2 [M] + .

[0244] Step 5: Preparation of the target product methyl l-(3-((5-amino-6-chloropyrimidin-4- yl)amino)-4-(4-methylpiperazin-l-yl)phenyl)-lH-l,2,3-triazole-4-carboxylate

[0245] Prepared according to the procedure of Example 23, Step 4, substituting 3'-((5-amino- 6-chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester for 3'-amino-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester, to give a white solid in 65.9% yield; m.p. 231-233 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.46 (s, 1H), 8.62 (s, 1H), 8.34 (s, 1H), 7.95 (s, 1H), 7.64 (d, J = 8.6 Hz, 1H), 7.37 (d, J = 8.7 Hz, 1H), 5.44 (s, 2H), 3.89 (s, 3H), 2.88 (s, 4H), 2.51 (s, 4H), 2.23 (s, 3H). HRMS (ESI): calcd. for m / z C 19 H 22 ClFN9O2,[M+H] + 444.1657, found 444.1660.HPLC (90% methanol in water): t R = 3.775 min, 97.63%.

[0246] Example 47

[0247]

[0248] (1-(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1-yl)phenyl)-1H- 1,2,3-triazol-4-yl)(morpholino)methanone

[0249] Step 1: Preparation of 1-(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4- methylpiperazin-1-yl)phenyl)-1H-1,2,3-triazole-4-carboxylic acid

[0250] Prepared according to the procedure of Example 23, Step 5, substituting 3'-((5-amino- 6-chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester for 1-(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4- methylpiperazin-1-yl)phenyl)-1H-1,2,3-triazole-4-carboxylic acid methyl ester, to give a white solid in 87.6% yield; m.p. 232-234 °C; 1H NMR (300 MHz, DMSO-d6) δ 12.20 (s, 1H), 9.36 (s, 1H), 8.62 (s, 1H), 8.34 (s, 1H), 7.95 (s, 1H), 7.63 (d, J = 8.7 Hz, 1H), 7.38 (d, J = 8.7 Hz, 1H), 5.47 (s, 2H), 2.90 (s, 4H), 2.53 (s, 4H), 2.34 (s, 3H). HRMS (ESI): calcd. for m / z C 18 H 20 ClFN9O2,[M+H] + 430.1488, found 430.1492.HPLC(90%methanol in water):t R = 3.548 min, 99.16%.

[0251] Step 2: Preparation of the title product (1-(3-((5-amino-6-chloropyrimidin-4- yl)amino)-4-(4-methylpiperazin-1-yl)phenyl)-1H-1,2,3-triazol-4-yl)(morpholino)methanone

[0252] Following the preparation procedure of Example 23, Step 6, replacing 3'-((5-amino-6- chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid with 1-(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1-yl)phenyl)-1H-1,2,3- triazol-4-carboxylic acid, afforded a white solid. Yield 68.3%; m.p. 200-202 °C; 1 H NMR (300 MHz, DMSO-d6) δ 12.20 (s, 1H), 9.36 (s, 1H), 8.62 (s, 1H), 8.34 (s, 1H), 7.95 (s, 1H), 7.63 (d, J = 8.7 Hz, 1H), 7.38 (d, J = 8.7 Hz, 1H), 5.47 (s, 2H), 2.90 (s, 4H), 2.53 (s, 4H), 2.34 (s, 3H). HRMS (ESI): calcd. for m / z C 24 H 32 ClFN 11 O2,[M+H] + 542.2502, found 542.2500.HPLC(80%methanol in water):tR = 4.612 min, 99.65 %.

[0253] Example 48

[0254]

[0255] 1 -(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin- 1 -yl)phenyl)-N-(3-morpholinopropyl)- 1 H- 1,2,3 -triazole-4-carboxamide

[0256] Following the procedure of Example 24, 3'-((5-amino-6-chloropyrimidin-4- yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1 '-biphenyl]-4-carboxylic acid was replaced by 1 -(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1- yl)phenyl)-1 H-1,2,3-triazole-4-carboxylic acid to give off-white solid. Yield 65.9 %; m.p. 203-205 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.19 (s, 1 H), 8.82 (t, J = 5.6 Hz, 1 H), 8.59 (s, 1 H), 8.34 (s, 1 H) 7.94 (s, 1 H), 7.62 (d, J = 8.7 Hz, 1 H), 7.36 (d, J = 8.7 Hz, 1 H), 5.45 (s, 2H), 3.61 (s, 4H), 3.36-3.33 (m, 4H), 2.90 (s, 4H), 2.58 (s, 4H), 2.40 (s, 4H), 2.28 (s, 3H), 1.73-1.68 (m, 2H). HRMS (ESI): calcd. for m / z C 25 H 34 ClFN 11 O2,[M+H] + 556.2660, found 556.2662. HPLC (80 % methanol in water): t R = 4.457 min, 98.27 %.

[0257] Example 49

[0258]

[0259] (1 -(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin- 1 - yl)phenyl)-1 H-1,2,3-triazol-4-yl)(4-methylpiperazin-1 -yl)methanone

[0260] Following the procedure of Example 25, 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid was replaced by 1-(3-((5-amino-6- chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1-yl)phenyl)-1H-1,2,3-triazole-4-carboxylic acid to give off-white solid. Yield 46.2%; m.p. 225-228 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.19 (s, 1H), 8.72 (s, 1H), 8.52 (s, 1H), 8.34 (s, 1H) 7.94 (s, 1H), 7.62 (d, J = 8.7 Hz, 1H), 7.37 (d, J = 8.7 Hz, 1H), 5.45 (s, 2H), 3.76 (s, 4H), 3.39 (s, 4H), 2.93 (s, 4H), 2.68 (s, 4H) 2.22 (s, 3H), 2.15 (s, 3H). HRMS (ESI): calcd. for m / z C 23 H 31 ClFN 11 O,[M+H] + 512.2396, found 512.2397.HPLC (90% methanol in water): t R = 3.707 min, 98.00%.

[0261] Example 50

[0262]

[0263] 1-(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1-yl)phenyl)-N,N- dimethyl-1H-1,2,3-triazole-4-carboxamide

[0264] Following the procedure of Example 27, 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid was replaced by 1-(3-((5-amino-6- chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1-yl)phenyl)-1H-1,2,3-triazole-4-carboxylic acid to give off-white solid. Yield 66.6%; m.p. 215-217 °C; 1H NMR (300 MHz, DMSO-d6) δ 9.84 (s, 1H), 9.18 (s, 1H), 8.62 (s, 1H), 8.27 (s, 1H) 7.95 (s, 1H), 7.68 (s, 1H), 7.44 (s, 1H), 5.52 (s, 2H), 3.58 (s, 4H), 3.30 (s, 6H), 3.04 (s, 4H), 2.88 (s, 3H). HRMS (ESI): calcd. for m / z C 20 H 25 ClFN 10 O,[M+H] + 457.1974, found 457.1966.HPLC (80% methanol in water): t R = 4.203 min, 98.52%.

[0265] Example 51

[0266]

[0267] 1-(3-((5-Amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1-yl)phenyl)-N,N- dimethyl-1H-1,2,3-triazole-4-carboxamide

[0268] Following the preparation procedure of Example 26, 3'-((5-amino-6-chloropyrimidin-4- yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid was replaced by 1- (3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1-yl)phenyl)-1H-1,2,3- triazole-4-carboxylic acid to give off-white solid. Yield 66.6%; m.p. 215 °C-217 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.84 (s, 1H), 9.18 (s, 1H), 8.62 (s, 1H), 8.27 (s, 1H) 7.95 (s, 1H), 7.68 (s, 1H), 7.44 (s, 1H), 5.52 (s, 2H), 3.58 (s, 4H), 3.30 (s, 6H), 3.04 (s, 4H), 2.88 (s, 3H). HRMS (ESI): calcd. for m / z C 20 H 25 ClFN 10 O,[M+H] + 457.1974, found 457.1966.HPLC (80% methanol in water): t R= 4.203 min, 98.52 %.

[0269] Example 52

[0270]

[0271] 1 -(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin- 1 -yl)phenyl)-N-(2-(dimethylamino)ethyl)- 1 H- 1,2,3 -triazole-4-carboxamide

[0272] Following the preparation procedure for Example 28, replace 3'-((5-amino-6- chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-l-yl)-[l,l'-biphenyl]-4-carboxylic acid with 1 -(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin- 1 -yl)phenyl)- 1 H- 1,2,3 -triazole-4-carboxylic acid to afford off-white solid. Yield 39.9 %; m.p. 149-152 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.84 (s, 1H), 9.18 (s, 1H), 8.62 (s, 1H), 8.54 (s, 1H), 7.95 (s, 1H), 7.68 (d, J = 8.7 Hz, 1H), 7.44 (d, J = 8.7 Hz, 1H), 5.46 (s, 2H), 3.08-2.90 (m, 6H), 2.77 (s, 6H), 2.50 (s, 4H), 1.90 (s, 4H), 1.28-1.23 (m, 2H). HRMS (ESI): calcd. for m / z C 22 H 30 ClFN 11 O, [M+H] + 500.2402, found 500.2404. HPLC (80 % methanol in water): t R = 4.157 min, 98.88 %.

[0273] Example 53

[0274]

[0275] 1 -(3-((5-amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin- 1 -yl)phenyl)-N-(2-(dimethylamino)ethyl)- 1 H- 1,2,3 -triazole-4-carboxamide

[0276] Following the procedure of Preparation 29, replace 3'-((5-amino-6-chloropyrimidin-4- yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid with 1-(3-((5- amino-6-chloropyrimidin-4-yl)amino)-4-(4-methylpiperazin-1-yl)phenyl)-1H-1,2,3- triazole-4-carboxylic acid to give a white solid. Yield 40.7%; m.p. 172-175 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.19 (s, 1H), 8.72 (s, 1H), 8.52 (s, 1H), 8.34 (s, 1H) 7.94 (s, 1H), 7.62 (d, J = 8.7 Hz, 1H), 7.37 (d, J = 8.7 Hz, 1H), 5.45 (s, 2H), 3.64-3.62 (m, 2H), 3.38-3.34 (m, 6H), 3.12 (s, 4H), 2.54 (s, 2H), 2.48 (s, 2H), 2.36 (s, 3H), 1.93 (s, 4H). HRMS (ESI): calcd. for m / z C 24 H 32 ClFN 11 O,[M+H] + 526.2553, found 526.2543.HPLC (80% methanol in water): t R = 4.211 min, 99.19%.

[0277] Example 54

[0278]

[0279] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-N,N-dimethyl-4"-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxamide

[0280] Step 1: Preparation of 1-bromo-2,4-difluoro-3-nitrobenzene:

[0281] A mixture solution of sulfuric acid (10 mL) and trifluoroacetic acid (50 mL) was prepared under ice-bath condition, 2,6-difluoronitrobenzene (5.0 g, 31.4 mmol) was added, N-bromosuccinimide (6.15 g, 34.5 mmol) was added portionwise slowly, the reaction was stirred at room temperature for 18 h, the reaction was poured into ice-water (100 mL), extracted with ethyl acetate (3 x 50 mL), the organic layer was washed with 2 M sodium hydroxide, dried over anhydrous sodium sulfate, filtered, and concentrated to give a red oil (6.6 g) in 90.0% yield; 1H NMR (300 MHz, CDC13) δ 7.78 (m, 1H), 7.28-7.16 (m, 1H). m / z (EI-MS): 237.9 [M] +

[0282] Step 2: Preparation of 2',4'-difluoro-3'-nitro-[l,l'-biphenyl]-4-carboxylic acid methyl ester:

[0283] The preparation method of Step 1 of Example 23 was followed by replacing 4- bromo 1-F nitrobenzene with 1-bromo-2,4-difluoro-3-nitrobenzene. Yield 65.0%. 1 H NMR (300 MHz, CDC13) δ 7.78 (m, 1H), 7.28-7.16 (m, 1H). m / z (EI-MS): 237.9 [M] + .

[0284] Step 3: Preparation of 2'-fluoro-4'-(4-methylpiperazin-l-yl)-3'-nitro-[l,l'- biphenyl]-4-carboxylic acid methyl ester:

[0285] The preparation method of Step 2 of Example 23 was followed by replacing 4'- fluoro-3'-nitro-[l,l'-biphenyl]-4-carboxylic acid methyl ester with 2',4'-difluoro-3'- nitro-[l,l'-biphenyl]-4-carboxylic acid methyl ester. Yield 88.2%; m.p. 112-115 °C; 1 H NMR (300 MHz, CDC13) δ 7.78 (m, 1H), 7.28-7.16 (m, 1H). m / z (EI-MS): 237.9 [M] + .

[0286] Step 4: Preparation of 3'-amino-2'-fluoro-4'-(4-methylpiperazin-l-yl)-[l,l'- biphenyl]-4-carboxylic acid methyl ester:

[0287] The preparation method of Step 3 of Example 23 was followed by replacing 4'- (4-methylpiperazin-l-yl)-3'-nitro-[l,l'-biphenyl]-4-carboxylic acid methyl ester with 2'-fluoro-4'-(4-methylpiperazin-l-yl)-3'-nitro-[l,l'-biphenyl]-4-carboxylic acid methyl ester. Yield 73.5%; m.p. 162-165 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.01 - 7.99 (m, 2H), 7.68 - 7.66 (m, 2H), 6.75 (dd, J = 7.5, 5.7 Hz, 1H), 6.58 (d, J = 7.5 Hz, 1H), 4.35 (s, 2H), 3.95 (s, 3H), 3.39 (t, J = 4.7 Hz, 4H), 2.54 (t, J = 4.7 Hz, 4H), 2.34 (s, 3H). m / z (EI-MS): 344.2 [M] + .

[0288] Step 5: Preparation of 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester:

[0289] Following the preparation procedure of Example 23, Step 4, 3'-amino-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester was replaced with 3'- amino-2'-fluoro-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester. Yield 55.7%; m.p. 222 °C - 225 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.01 - 7.99 (m, 2H), 7.68 - 7.66 (m, 2H), 6.75 (dd, J = 7.5, 5.7 Hz, 1H), 6.58 (d, J = 7.5 Hz, 1H), 4.35 (s, 2H), 3.95 (s, 3H), 3.39 (t, J = 4.7 Hz, 4H), 2.54 (t, J = 4.7 Hz, 4H), 2.34 (s, 3H). m / z (EI-MS): 344.2 [M] 23 H 24 ClFN6O2,[M+H] + 471.1706, found 471.1707. HPLC (80% methanol in water): t R = 2.744 min, 99.52%.

[0290] Step 6: Preparation of 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid:

[0291] The preparation method according to the preparation method of Example 23, Step 5 was replaced with 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester. Two step yield 80.4%; m.p. >250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.11 (dd, J = 5.3, 2.2 Hz, 3H), 7.80 (s, 1H), 7.69 - 7.67 (m, 2H), 7.12 (dd, J = 7.5, 5.7 Hz, 1H), 6.66 (d, J = 7.5 Hz, 1H), 5.35 (s, 2H), 3.93 (s, 3H), 3.34 (t, J = 4.7 Hz, 4H), 2.57 (s, 4H), 2.32 (s, 3H). m / z (EI-MS): 457.2 [M] + .

[0292] Step 7: Preparation of the target product 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-N,N-dimethyl-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxamide:

[0293] Preparation method according to the similar preparation method of Example 23, Step 6. Yield 77.5%; m.p. 158 °C - 160 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.03 (s, 1H), 7.71 (s, 1H), 7.60 - 7.46 (m, 4H), 7.40 (t, J = 8.5 Hz, 1H), 7.00 (d, J = 8.6 Hz, 1H), 5.49 (s, 2H), 2.99 - 2.98 (m, 6H), 2.87 (s, 4H), 2.38 (s, 4H), 2.18 (s, 3H). HRMS (ESI): calcd. for m / z C 24 H 27 ClFN7O,[M+H] + 484.2202, found 484.2203. HPLC (80% methanol in water): t R = 3.448 min, 95.30%.

[0294] Example 55:

[0295]

[0296] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)- [1,1'-biphenyl]-4-yl)(morpholino)methanone

[0297] Following the preparation procedure of Example 54, dimethylamine was replaced by morpholine, yield 84.7%; m.p. 145-148°C; 1 H NMR (300 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.72 (s, 1H), 7.54 (dd, J = 8.5, 5.4 Hz, 4H), 7.41 (t, J = 8.5 Hz, 1H), 7.03 (d, J = 8.6 Hz, 1H), 5.48 (s, 2H), 3.61 (s, 4H), 3.34 (s, 4H), 2.98 (s, 4H), 2.65 (s, 4H), 2.38 (s, 3H). HRMS (ESI): calcd. for m / z C 26 H 29 ClFN7O2, [M+H] + 526.2128, found 526.2122. HPLC (80% methanol in water): t R = 3.560 min, 98.66%

[0298] Example 56

[0299]

[0300] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)-N-(2- morpholinoethyl)-[1,1'-biphenyl]-4-carboxamide

[0301] Following the preparation procedure of Example 54, dimethylamine was replaced by 2-morpholinoethyl-1-amine, yield 74.9%; m.p. 199-202°C; 1 H NMR (300 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.72 (s, 1H), 7.54 (dd, J = 8.5, 5.4 Hz, 4H), 7.41 (t, J = 8.5 Hz, 1H), 7.03 (d, J = 8.6 Hz, 1H), 5.48 (s, 2H), 3.61 (s, 4H), 3.34 (s, 4H), 2.98 (s, 4H), 2.65 (s, 4H), 2.38 (s, 3H). HRMS (ESI): calcd. for m / z C28 H 34 ClFN8O2,[M+H] + 569.2550, found 569.2548.HPLC(80%methanol in water):t R = 3.634 min, 98.88 %.

[0302] Example 57

[0303]

[0304] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)-N-(3-morpholinopropyl)-[1,1'-biphenyl]-4-carboxamide

[0305] According to the preparation method of Example 54, dimethylamine was replaced by 3-morpholinopropyl-1-amine, yield 65.2%; m.p. 202-205 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.46 (t, J = 5.7 Hz, 1H), 7.96 (s, 1H), 7.81 (d, J = 8.2 Hz, 2H), 7.31 (t, J = 8.6 Hz, 2H), 6.90 (d, J = 8.7 Hz, 1H), 5.40 (s, 2H), 3.47 (t, J = 5.4 Hz, 4H), 3.21-3.19 (m, 2H), 2.80 (s, 4H), 2.28-2.22 (m, 10H), 2.09 (s, 3H), 1.60 (t, J = 7.1 Hz, 2H). HRMS (ESI): calcd. for m / z C 29 H 37 ClFN8O2,[M+H] + 583.2706, found 583.2707.HPLC(80%methanol in water):t R = 3.601 min, 97.46 %.

[0306] Example 58

[0307]

[0308] (3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)(4-ethylpiperazin-1-yl)methanone

[0309] Preparation according to the procedure of Example 54, replacing dimethylamine with 1- ethylpiperazine, in 64.8% yield; m.p. 229-232 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.71 (s, 1H), 7.57 (d, J = 8.2 Hz, 2H), 7.47 (d, J = 8.1 Hz, 2H), 7.39 (t, J = 8.5 Hz, 2H), 7.00 (d, J = 8.7 Hz, 1H), 5.49 (s, 2H), 3.61 (s, 2H), 2.91 (s, 4H), 2.38-2.34 (m, 10H), 2.20 (s, 3H), 1.17 (t, J = 7.1 Hz, 2H), 1.01 (t, J = 7.1 Hz, 2H). HRMS (ESI): calcd. for m / z C 28 H 34 ClFN8O,[M+H] + 553.2587, found 553.2596.HPLC (80% methanol in water): t R = 3.794 min, 97.08%.

[0310] Example 59

[0311]

[0312] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)tetrahydro-2H-pyran-4-carboxamide

[0313] Step 1: Preparation of 2',4'-difluoro-3'-nitro[1,1'-biphenyl]-4-amine

[0314] Preparation according to the procedure of Example 23, Step 1, replacing 4-bromo 1-F nitrobenzene with 1-bromo-2,4-difluoro-3-nitrobenzene, in 69.0% yield; m.p. 136-138 °C; 1 H NMR (300 MHz, DMSO-d6) δ 7.87-7.83 (m, 1H), 7.24-7.22 (m, 2H), 7.15 (dd, J = 9.0, 7.5 Hz, 1H), 6.72-6.70 (m, 2H), 5.24 (s, 2H). m / z (EI-MS): 251.1 [M] + .

[0315] Step 2: Preparation of 2'-fluoro-4'-(4-methylpiperazin-1-yl)-3'-nitro[1,1'-biphenyl]-4-amine

[0316] Following the procedure for the preparation of Example 23, Step 2, replace 4'-fluoro-3'-nitro-[1,1'-biphenyl]-4-carboxylic acid methyl ester with 2',4'-difluoro-3'-nitro[1,1'-biphenyl]-4-amine to yield 88.2%; m.p. 145-147°C; 1 H NMR (300 MHz, DMSO-d6) δ 7.60 (dd, J = 7.4, 5.8 Hz, 1H), 7.25 - 7.18 (m, 2H), 6.96 (d, J = 7.5 Hz, 1H), 6.77 - 6.71 (m, 2H), 5.24 (s, 2H), 3.20 (t, J = 4.6 Hz, 4H), 2.54 (t, J = 4.6 Hz, 4H), 2.34 (s, 3H). m / z (EI-MS): 331.2 [M] + .

[0317] Step 3: Preparation of N-(2'-fluoro-4'-(4-methylpiperazin-1-yl)-3'-nitro[1,1'-biphenyl]-4-yl)tetrahydro-2H-pyran-4-carboxamide:

[0318] Following the procedure for the preparation of Example 33, replace N 4 -(4'-amino-4-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-3-yl)-6-chloropyrimidine-e-4,5-diamine with 2'-fluoro-4'-(4-methylpiperazin-1-yl)-3'-nitro[1,1'-biphenyl]-4-amine to yield 88.2%; m.p. 166-168°C; 1 H NMR (300 MHz, DMSO-d6) δ 9.52 (s, 1H), 7.63 (dd, J = 7.5, 5.7 Hz, 1H), 7.42 - 7.38 (m, 4H), 6.99 (d, J = 7.5 Hz, 1H), 3.95 - 3.92 (m, 2H), 3.53 - 3.50 (m, 2H), 3.20 (t, J = 4.7 Hz, 4H), 2.54 (t, J = 4.7 Hz, 4H), 2.65 - 2.60 (m, 4H), 2.08 - 2.01 (m, 2H), 1.83 - 1.76 (m, 2H). m / z (EI-MS): 443.2 [M] + .

[0319] Step 4: Preparation of the target product N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"- (4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)tetrahydro-2H-pyran-4-carboxamide

[0320] Compound 59 was prepared in 36.6% overall yield in three steps according to the similar procedures described in Example 23, Step 3 and Step 4; m.p. >250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.56 (s, 1H), 8.25 (s, 1H), 7.80-7.77 (m, 3H), 7.41-7.39 (m, 2H), 7.07 (dd, J = 7.5, 5.7 Hz, 1H), 6.68 (d, J = 7.3 Hz, 1H), 5.43 (s, 2H), 3.52-3.49 (m, 4H), 3.34 (t, J = 4.6 Hz, 4H), 2.70-2.67 (m, 1H), 2.44 (s, 4H), 2.22 (s, 3H), 2.06-1.99 (m, 2H), 1.85-1.78 (m, 2H).. HRMS (ESI): calcd. for m / z C 27 H 31 ClFN7O2,[M+H] + 540.2285, found 540.2276.HPLC (90% methanol in water): t R = 3.656 min, 98.94%.

[0321] Example 60

[0322]

[0323] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)- [1,1'-biphenyl]-pyridin-4-yl)-1-methylpiperidine-4-carboxamide

[0324] Step 1: Preparation of N-(2'-fluoro-4'-(4-methylpiperazin-1-yl)-3'-nitro[1,1'-biphenyl]-4-yl)-1- methylpiperidine-4-carboxamide

[0325] According to the preparation procedure of Example 59, Step 3, replace tetrahydropyran-4- carboxylic acid with 1-methylpiperazine-4-carboxylic acid, in 75.3% yield; m.p. 170-172 °C; 1H NMR (300 MHz, DMSO-d6) δ 9.55 (s, 1H), 7.63 (dd, J = 7.5, 5.7 Hz, 1H), 7.40 (s, 4H), 6.98 (d, J = 7.5 Hz, 1H), 3.22 (t, J = 4.7 Hz, 4H), 2.99-2.93 (m, 6H), 2.64-2.58 (m, 4H), 2.37 (s, 3H), 2.29-2.24 (m, 2H), 2.09-2.02 (m, 2H), 1.71-1.64 (m, 2H). m / z (EI-MS): 455.2 [M] + .

[0326] Step 2: Preparation of the target compound N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-l-yl)-[l,l'-biphenyl]-pyridin-4-yl)-l- methylpiperidine-4-carboxamide:

[0327] According to the similar preparation method of Example 23, Step 3 and Step 4, compound 60 was obtained in 35.3% yield over three steps; m.p. > 250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.66 (s, 1H), 8.18 (s, 1H), 7.80 (s, 1H), 7.63 (d, J = 7.5, 2H), 7.40 (d, J = 7.6, 2H), 7.08-7.05 (m, 1H), 6.61 (d, J = 7.5 Hz, 1H), 5.30 (s, 2H), 3.20 (t, J = 4.9 Hz, 4H), 3.01-2.96 (m, 6H), 2.63-2.60 (m, 4H), 2.37 (s, 3H), 2.21-2.18 (m, 4H), 2.07-2.02 (m, 4H). HRMS (ESI): calcd. for m / z C 28 H 34 ClFN8O,[M+H] + 553.2607, found 553.2606. HPLC (80% methanol in water): t R = 3.168 min, 97.33%.

[0328] Example 61

[0329]

[0330] N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin- 1-yl)-[1,1'-biphenyl]-piperidin-4-yl)-2-(tetrahydro-2H-pyran-4-yl)acetamide

[0331] Step 1: Preparation of N-(2'-fluoro-4'-(4-methylpiperazin-1-yl)-3'-nitro[1,1'-biphenyl]- 4-yl)-2-(tetrahydro-2H-pyran-4-yl)acetamide

[0332] According to the similar preparation method of Example 59 Step 3, tetrahydro- pyran-4-carboxylic acid was replaced by tetrahydro-pyran-4-acetic acid, yield 68.9%; m.p. 148-150 °C; 1 H NMR (300 MHz, DMSO-d6) δ 7.80-7.77 (m, 2H), 7.70 (s, 1H), 7.65-7.63 (m, 1H), 7.39 (d, J = 7.5 Hz, 2H), 6.94 (d, J = 7.5 Hz, 1H), 3.90-3.87 (m, 2H), 3.55-3.51 (m, 2H), 3.24 (t, J = 4.7 Hz, 4H),, 2.60 (s, 3H), 2.38 (t, J = 4.7 Hz, 4H), 2.15 (d, J = 7.0 Hz, 2H), 2.05-1.99 (m, 1H), 1.20-1.17 (m, 4H). m / z (EI-MS): 457.2 [M] + .

[0333] Step 2: Preparation of the target product N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-piperidin-4-yl)-2-(tetrahydro- 2H-pyran-4-yl)acetamide

[0334] According to the similar preparation method of Example 23 Step 3 and Step 4, compound 61 was obtained, three-step yield 38.6%; m.p. >250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.24 (s, 1H), 8.11 (s, 1H), 7.80 (s, 1H), 7.40 (s, 4H), 7.15-7.12 (m, 1H), 6.73 (d, J = 7.5 Hz, 1H), 5.42 (s, 2H), 3.55-3.50 (m, 4H), 3.53 (t, J = 4.6 Hz, 4H), 2.48 (s, 4H), 2.26 (s, 3H), 2.15 (d, J = 7.0 Hz, 2H), 2.03-2.01 (m, 1H), 1.70-1.64 (m, 4H). HRMS (ESI): calcd. for m / z C28 H 33 ClFN7O2,[M+H] + 554.2441,found 554.2440.HPLC(80%methanol in water):t R = 3.524 min, 99.10%. Example 62

[0335] Example 62

[0336]

[0337] 2-Amino-N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)-3-methylbutanamide

[0338] Step 1: Preparation of (1-((2'-fluoro-4'-(4-methylpiperazin-1-yl)-3'-nitro-[1,1'- biphenyl]-4-yl)amino)-3-methyl-tert-butyl-1-oxobutan-2-yl)carbamic acid

[0339] According to the similar preparation method of Example 59 Step 3, replace tetrahydro- pyran-4-carboxylic acid with (tert-butoxycarbonyl)valine, yield 69.0%; m.p. 108-110 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.63 (s, 1H), 7.64-7.61 (m, 3H), 7.44-7.42 (m, 2H), 7.39 (s, 3H), 6.98 (d, J = 7.5 Hz, 1H), 4.13-4.12 (m, 1H), 3.75-3.72 (m, 2H), 3.31-3.27 (m, 2H), 2.79-2.77 (m, 2H), 2.63-2.61 (m, 1H), 2.60 (s, 3H), 2.24-2.22 (m, 2H), 1.44 (s, 9H), 0.94-0.88 (m, 6H). m / z (EI-MS): 530.3 [M] + .

[0340] Step 2: Preparation of the target product 2-amino-N-(3"-((5-amino-6-chloropyrimidin-4- yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)-3-methylbutanamide

[0341] According to the similar preparation method of Example 23 Step 3 and Step 4, compound 62 was obtained in three steps with a yield of 46.9%, m.p. 189-192 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.74-7.71 (m, 3H), 7.45 (s, 2H), 7.33 (t, J = 8.5 Hz, 1H), 6.96 (d, J = 7.2 Hz, 1H), 5.47 (s, 2H), 3.15-3.12 (m, 1H), 2.86 (s, 4H), 2.36 (s, 4H), 2.16 (s, 3H), 1.95 (s, 2H), 1.17 (s, 1H), 0.93 (s, 3H), 0.86 (s, 3H). HRMS (ESI): calcd. for m / z C 26 H 32 ClFN8O,[M+H] + 527.2444, found 527.2449.HPLC (80% methanol in water): t R = 3.858 min, 99.62%.

[0342] Example 63

[0343]

[0344] 2-Amino-N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)-4-methylpentanamide

[0345] Step 1: Preparation of (1-((2'-fluoro-4'-(4-methylpiperazin-1-yl)-3'-nitro-[1,1'- biphenyl]-4-yl)amino)-4-methyl-tert-butyl-1-oxopentan-2-yl)carbamic acid

[0346] According to Example 59, Step 3, replace tetrahydropyran-4-carboxylic acid with (tert-butoxycarbonyl)leucine, yield 67.2%; m.p. 114 °C - 116 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.46 (s, 1H), 7.65-7.62 (m, 1H), 7.45-7.42 (m, 4H), 6.95 (d, J = 7.5 Hz, 1H), 6.24 (s, 1H), 4.04-4.01 (m, 1H), 3.87-3.82 (m, 2H), 3.31-3.25 (m, 2H), 2.79-2.74 (m, 2H), 2.25-2.19 (m, 4H), 1.98-1.92 (m, 2H), 1.71-1.65 (m, 1H), 1.44 (s, 9H), 0.96-0.90 (m, 6H). m / z (EI-MS): 544.3 [M] + .

[0347] Step 2: Preparation of the title product 2-amino-N-(3"-((5-amino-6-chloropyrimidin-4- yl)amino)-2'-fluoro-4"-(4-methylpiperazin-l-yl)-[l,l'-biphenyl]-4-yl)-4- methylpentanamide

[0348] Following a similar preparation procedure as in Example 23, Step 3 and Step 4, compound 63 was obtained in 47.6% yield over three steps; m.p. 182-185 °C; 1 H NMR (300 MHz, DMSO-d6) δ 7.99 (s, 1H), 7.73-7.71 (m, 3H), 7.45 (d, J = 8.1 Hz, 2H), 7.33 (t, J = 8.5 Hz, 2H), 6.96 (d, J = 8.6 Hz, 2H), 5.45 (s, 2H), 3.38-3.36 (m, 1H), 2.87 (s, 4H), 2.36 (s, 4H), 2.17 (s, 3H), 1.78-1.74 (m, 2H), 1.52-1.47 (m, 2H), 1.19-1.17 (m, 1H), 0.90 (t, J = 7.2 Hz, 6H). m / z (EI-MS): 541.3 [M] + HPLC (80% methanol in water): t R = 3.680 min, 98.32%.

[0349] Example 64

[0350]

[0351] 2-amino-N-(3"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4- methylpiperazin-l-yl)-[l,l'-biphenyl]-4-yl)acetamide

[0352] Step 1: Preparation of (2-((2'-fluoro-4'-(4-methylpiperazin-l-yl)-3'-nitro-[l,l'-biphenyl]-4- yl)amino)-2-oxoethyl tert-butyl)carbamate

[0353] Following a similar preparation procedure as in Example 59, Step 3, replacing tetrahydropyran-4-carboxylic acid with (tert-butoxycarbonyl)glycine, in 70.0% yield; m.p. 112-114 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.20 (s, 1H), 7.63 (dd, J = 7.5, 5.7 Hz, 1H), 7.49-7.47 (m, 2H), 7.44-7.42 (m, 2H), 6.94 (d, J = 7.5 Hz, 1H), 6.31 (s, 1H), 3.89 (s, 2H), 3.30 (t, J = 4.7 Hz, 4H), 2.44 (s, 4H), 2.24 (s, 3H), 1.44 (s, 9H). m / z (EI-MS): 486.2 [M] + .

[0354] Step 2: Preparation of the title compound 2-amino-N-(3"-((5-amino-6- chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4- yl)acetamide

[0355] Following the similar preparation procedure as in Example 23, Step 3 and Step 4, compound 64 was obtained in 50.9% yield over three steps; m.p. 177 °C-179 °C; 1 H NMR (300 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.63-7.61 (m, 3H), 7.36 (d, J = 8.2 Hz, 2H), 7.24 (t, J = 8.5 Hz, 2H), 6.87 (d, J = 8.5 Hz, 2H), 5.35 (s, 2H), 4.00 (s, 1H), 3.08 (s, 2H), 2.77 (t, J = 4.7 Hz, 4H), 2.27 (t, J = 4.6 Hz, 4H), 2.07 (s, 3H). m / z (EI-MS): 485.3 [M] + HPLC (80% methanol in water): t R = 3.595 min, 99.76%.

[0356] Example 65

[0357]

[0358] 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-N,N,2'-trimethyl-4"-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxamide

[0359] Step 1: Preparation of 1-bromo-4-fluoro-2-methyl-3-nitrobenzene

[0360] Following the preparation procedure of Example 54, Step 1, replace 2,6- difluoronitrobenzene with 1-bromo-2,4-difluoro-3-nitrobenzene, yield 86.7%; m.p. 74 °C-77 °C;1 H NMR (300 MHz, CDC13) δ 7.72 (d, J = 7.5, Hz, 1 H), 7.03 (d, J = 7.5 Hz, 1 H), 2.32 (s, 3 H). m / z (EI-MS): 233.9 [M] +

[0361] Step 2: Preparation of 4'-fluoro-N,N,2'-trimethyl-3'-nitro-[1,1'-biphenyl]-4- carboxamide

[0362] Following the preparation procedure of Example 54, Step 2, replace (4- (methoxycarbonyl)phenyl)boronic acid with (4-(dimethylcarbamoyl)phenyl)boronic acid, yield 76.9 %; 1 H NMR (300 MHz, CDC13) δ 7.72 (d, J = 7.5, Hz, 1 H), 7.03 (d, J = 7.5 Hz, 1 H), 2.32 (s, 3 H). m / z (EI-MS): 233.9 [M] + .

[0363] Step 3: Preparation of N,N,2'-trimethyl-4'-(4-methylpiperazin-1-yl)-3'-nitro[1,1'- biphenyl]-4-carboxamide

[0364] Following the preparation procedure of Example 54, Step 3, replace 4'-fluoro- 3'-nitro-[1,1'-biphenyl]-4-carboxylic acid methyl ester with 4'-fluoro-N,N,2'-trimethyl- 3'-nitro-[1,1'-biphenyl]-4-carboxamide. Yield 85.5 %; m.p. 110-112 °C; 1 H NMR (300 MHz, CDC13) δ 7.72 (d, J = 7.5, Hz, 1 H), 7.03 (d, J = 7.5 Hz, 1 H), 2.32 (s, 3 H). m / z (EI-MS): 233.9 [M] + .

[0365] Step 4: Preparation of 3'-amino-N,N,2'-trimethyl-4'-(4-methylpiperazin-1-yl)- [1,1'-biphenyl]-4-carboxamide:

[0366] Preparation according to the procedure of Example 54, step 4, substituting 4'-(4- methylpiperazin-1-yl)-3'-nitro-[1,1'-biphenyl]-4-carboxylic acid methyl ester with N,N,2'-trimethyl-4'-(4-methylpiperazin-1-yl)-3'-nitro[1,1'-biphenyl]-4-carboxamide. Yield 80.5%; m.p. 158-160 °C; 1 H NMR (300 MHz, DMSO-d6) δ 7.91-7.89 (m, 2H), 7.61-7.59 (m, 2H), 6.68-6.64 (m, 2H), 4.35 (s, 2H), 3.39 (t, J = 4.7 Hz, 4H), 3.03 (s, 6H), 2.54 (s, 4H), 2.14 (s, 3H), 2.09 (s, 3H). m / z (EI-MS): 353.2 [M] + .

[0367] Step 5: Preparation of the target product 3"-((5-amino-6-chloropyrimidin-4-yl)amino)-N,N,2'-trimethyl-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxamide

[0368] Preparation according to the procedure of Example 54, step 5, to give compound 65. Yield 44.5%; m.p. 165-167 °C; 1 H NMR (300 MHz, DMSO-d6) δ 7.91-7.89 (m, 2H), 7.61-7.59 (m, 2H), 6.68-6.64 (m, 2H), 4.35 (s, 2H), 3.39 (t, J = 4.7 Hz, 4H), 3.03 (s, 6H), 2.54 (s, 4H), 2.14 (s, 3H), 2.09 (s, 3H). m / z (EI-MS): 353.2 [M] + HPLC (80% methanol in water): t R = 3.543 min, 96.37%.

[0369] Example 66

[0370]

[0371] Step 1: Preparation of 2',4'-difluoro-5'-nitro-[1,1'-biphenyl]-4-carboxylic acid methyl ester

[0372] Preparation according to the procedure of Example 23, Step 1, substituting 4-bromo 1-F nitrobenzene for 1-bromo-2,4-difluoro-5-nitrobenzene. Yield 67.8%; m.p. 134-136 °C; 1 H NMR (300 MHz, CDC13) δ 8.30 (t, J = 5.7 Hz, 1H), 7.97 (d, J = 8.9 Hz, 2H), 7.57 (d, J = 5.8 Hz, 2H), 7.24 (t, J = 8.9 Hz, 1H), 3.95 (s, 3H). m / z (EI-MS): 294.1 [M] + .

[0373] Step 2: Preparation of 2'-fluoro-4'-(4-methylpiperazin-l-yl)-5'-nitro-[l,l'-biphenyl]-4- carboxylic acid methyl ester

[0374] Preparation according to the procedure of Example 23, Step 2, substituting 4'-fluoro-3'- nitro-[l,l'-biphenyl]-4-carboxylic acid methyl ester for 2',4'-difluoro-5'-nitro-[l,l'- biphenyl]-4-carboxylic acid methyl ester. Yield 84.3%; m.p. 137-140 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.02-7.99 (m, 3H), 7.68-7.65 (m, 2H), 6.79 (d, J = 7.5 Hz, 1H), 3.95 (s, 3H), 3.39 (t, J = 4.7 Hz, 4H), 2.54 (t, J = 4.6 Hz, 4H), 2.14 (s, 3H). m / z (EI-MS): 374.1 [M] + .

[0375] Step 3: Preparation of 5'-amino-2'-fluoro-4'-(4-methylpiperazin-l-yl)-[l,l'-biphenyl]-4- carboxylic acid methyl ester

[0376] Preparation according to the procedure of Example 23, Step 3, substituting 4'-(4- methylpiperazin-l-yl)-3'-nitro-[l,l'-biphenyl]-4-carboxylic acid methyl ester for 2'- fluoro-4'-(4-methylpiperazin-l-yl)-5'-nitro-[l,l'-biphenyl]-4-carboxylic acid methyl ester. Yield 80.6%; m.p. 162-165 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.00 (d, J = 8.9 Hz, 2H), 7.68 (d, J = 8.8 Hz, 2H), 6.56 (d, J = 5.7 Hz, 1H), 6.39 (d, J = 8.9 Hz, 1H), 4.35 (s, 2H), 4.35 (s, 2H), 3.95 (s, 3H), 3.39 (t, J = 4.7 Hz, 4H), 2.54 (t, J = 4.7 Hz, 4H), 2.17 (s, 3H). m / z (EI-MS): 344.2 [M] + .

[0377] Step 4: Preparation of the title compound methyl 5'-((5-amino-6-chloropyrimidin-4- yl)amino)-2'-fluoro-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylate:

[0378] According to the similar preparation method of Example 23, Step 4, compound 66 was obtained in a yield of 57.4%; m.p. 220-223 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.00 (d, J = 8.9 Hz, 2H), 7.68 (d, J = 8.8 Hz, 2H), 6.56 (d, J = 5.7 Hz, 1H), 6.39 (d, J = 8.9 Hz, 1H), 4.35 (s, 2H), 4.35 (s, 2H), 3.95 (s, 3H), 3.39 (t, J = 4.7 Hz, 4H), 2.54 (t, J = 4.7 Hz, 4H), 2.17 (s, 3H). m / z (EI-MS): 344.2 [M] 23 H 24 ClFN6O2, [M+H] + 471.1706, found 471.1706. HPLC (80% methanol in water): t R = 4.248 min, 98.83%.

[0379] Example 67

[0380]

[0381] (5"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1- yl)-[1,1'-biphenyl]-4-yl)(morpholino)methanone

[0382] Step 1: Preparation of 5"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid

[0383] The procedure of Example 23, Step 5 was followed to replace 3'-((5-amino-6- chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester with 5'-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester. Yield 82.7%; m.p. >250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.14 - 8.10 (m, 3H), 7.80 (s, 1H), 7.73 - 7.70 (m, 2H), 6.80 (d, J = 5.7 Hz, 1H), 6.57 (d, J = 8.9 Hz, 1H), 5.34 (s, 2H), 3.93 (s, 3H), 3.34 (t, J = 4.7 Hz, 4H), 2.57 (s, 4H), 2.12 (s, 3H). m / z (EI-MS): 457.2 [M] + .

[0384] Step 2: Preparation of the target product (5"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'- fluoro-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)(morpholino)methanone

[0385] The procedure of Example 23, Step 6 was followed to replace 3'-((5-amino-6- chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester with 5'-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester to obtain the target compound 67. Yield 78.4%; m.p. 136 °C - 138 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.14 - 8.10 (m, 3H), 7.80 (s, 1H), 7.73 - 7.70 (m, 2H), 6.80 (d, J = 5.7 Hz, 1H), 6.57 (d, J = 8.9 Hz, 1H), 5.34 (s, 2H), 3.93 (s, 3H), 3.34 (t, J = 4.7 Hz, 4H), 2.57 (s, 4H), 2.12 (s, 3H). m / z (EI-MS): 457.2 [M] 26 H 29 ClFN7O2,[M+H] + 526.2128, found 526.2130.HPLC (80% methanol in water): t R = 4.046 min, 96.85%.

[0386] Example 68

[0387]

[0388] 5'-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)-N-(2-morpholinoethyl)-[1,1' :1''-terphenyl]-4-carboxamide

[0389] The target compound 68 was obtained by the similar preparation method as in Example 67, replacing morpholine with 1-ethylmorpholine. Yield 78.4%; m.p. 199-202°C; 1 H NMR (300 MHz, DMSO-d6) δ 8.47 (s, 1H), 8.09 (s, 1H), 7.95-7.92 (m, 3H), 7.81 (s, 1H), 7.62 (d, J = 7.9 Hz, 2H), 7.11 (d, J = 12.4 Hz, 1H), 5.39 (s, 2H), 3.58 (t, J = 4.6 Hz, 4H), 3.41-3.40 (m, 4H), 2.92 (s, 4H), 2.56 (s, 4H), 2.45 (s, 4H), 2.29 (s, 3H). HRMS (ESI): calcd. for m / z C 28 H 34 ClFN8O2, [M+H] + 569.2550, found 569.2548. HPLC (80% methanol in water): t R = 4.049 min, 98.64%.

[0390] Example 69

[0391]

[0392] 5'-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin-1-yl)-N-(3-morpholinopropyl)-[1,1' :1''-terphenyl]-4-carboxamide

[0393] The target compound 69 was obtained by the similar preparation method as in Example 67, replacing morpholine with 1-propylmorpholine. Yield 78.4%; m.p. 204-206°C; 1H NMR (300 MHz, DMSO-d6) δ 8.55 (s, 1H), 8.09 (s, 1H), 7.93-7.90 (m, 3H), 7.81 (s, 1H), 7.63-7.60 (m, 2H), 7.11 (d, J = 12.3 Hz, 1H), 5.39 (s, 2H), 3.59 (s, 4H), 3.32 (s, 4H), 2.92 (s, 4H), 2.58 (s, 4H), 2.41 (s, 4H), 2.31 (s, 3H), 1.72 (s, 2H). HRMS (ESI): calcd. for m / z C 29 H 36 ClFN8O2, [M+H] + 583.2693, found 583.2699. HPLC (80% methanol in water): t R = 3.885 min, 99.38%.

[0394] Example 70

[0395]

[0396] 5"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-N-(4- hydroxycyclohexyl)-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxamide

[0397] According to the similar preparation method of Example 67, replacing morpholine with piperidin-4-ol to obtain the target compound 70. Yield 65.2%; m.p. 200-203 °C; 1 H NMR (300 MHz, DMSO-d6) δ 9.67 (s, 1H), 8.06 (s, 1H), 7.92 (d, J = 8.4 Hz, 1H), 7.82 (s, 1H), 7.58 (d, J = 7.7 Hz, 2H), 7.47 (d, J = 7.9 Hz, 2H), 7.21 (d, J = 12.2 Hz, 1H), 5.44 (s, 2H), 4.82 (s, 1H), 4.01 (s, 2H), 3.52 (s, 2H), 3.16 (s, 8H), 2.78 (s, 3H), 1.76 (s, 2H), 1.36 (s, 2H). HRMS (ESI): calcd. for m / z C 27 H 31 ClFN7O2, [M+H] + 554.2441, found 554.2442. HPLC (80% methanol in water): t R= 4.057 min, 98.48 %.

[0398] Example 71

[0399]

[0400] 5'-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-4"-(4-methylpiperazin- 1 -yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)-[1, 1 '-biphenyl]-4-carboxamide

[0401] Following a similar preparation method as Example 67, replacing morpholine with (tetrahydro-2H-pyran-4-yl)methanamine, the target compound 71 was obtained. Yield 75.2 %; m.p. 189-191 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.10 (s, 1H), 7.94-7, 91 (m, 3H), 7.61 (d, J = 8.0 Hz, 2H), 7.11 (d, J = 12.2 Hz, 1H), 5.40 (s, 2H), 3.87-3.83 (m, 2H), 3.17 (s, 4H), 2.92 (s, 4H), 2.57 (s, 4H), 2.30 (s, 3H), 1.81 (s, 1H), 1.23 (s, 4H). HRMS (ESI): calcd. for m / z C 28 H 33 ClFN7O2, [M+H] + 540.2285, found 540.2284. HPLC (100 % methanol): t R = 7.989 min, 99.16 %.

[0402] Example 72

[0403]

[0404] 5'-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-fluoro-N,N-dimethyl-4"-(4- methylpiperazin-1 -yl)-[1, 1 '-biphenyl]-4-carboxamide

[0405] Following a similar preparation method as Example 67, replacing morpholine with dimethylamine, the target compound 72 was obtained. Yield 75.2 %; m.p. 157-159 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.01 (s, 1H), 7.83 (dd, J = 8.7, 3.7 Hz, 1H), 7.72 (d, J = 3.6 Hz, 1H), 7.49 (d, J = 7.1 Hz, 2H), 7.45 - 7.36 (m, 2H), 7.14 - 7.01 (m, 1H), 5.35 (s, 2H), 3.27-2.87 (m, 14H), 2.40 (s, 3H). HRMS (ESI): calcd. for m / z C 24 H 27 ClFN7O,[M+H] + 484.2022, found 484.2022.HPLC (80% methanol in water): t R = 4.751 min, 95.34%.

[0406] Example 73

[0407]

[0408] 5'-((5-Amino-6-chloropyrimidin-4-yl)amino)-2'-methyl-4'-(4-methylpiperazin-1-yl)- [1,1'-biphenyl]-4-carboxylic acid methyl ester Step 1: Preparation of 4'-fluoro-2'-methyl-5'-nitro-[1,1'-biphenyl]-4-carboxylic acid methyl ester

[0409] The preparation method of Example 23, Step 1 was followed by replacing 4-bromo 1-F nitrobenzene with 1-bromo-4-fluoro-2-methyl-5-nitrobenzene. Yield 68.9%; m.p. 133 °C - 134 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.06 (dd, J = 9.4, 3.2 Hz, 2H), 7.97 (d, J = 7.7 Hz, 1H), 7.62 - 7.57 (m, 3H), 3.89 (s, 3H), 2.33 (s, 3H). m / z (EI-MS): 290.1 [M] + .

[0410] Step 2: Preparation of 2'-methyl-4'-(4-methylpiperazin-1-yl)-5'-nitro-[1,1'-biphenyl]-4- carboxylic acid methyl ester

[0411] The preparation method of Example 23, Step 2 was followed by replacing 4'-fluoro-3'-nitro- [1,1'-biphenyl]-4-carboxylic acid methyl ester with 4'-fluoro-2'-methyl-5'-nitro-[1,1'- biphenyl]-4-carboxylic acid methyl ester. Yield 84.3%; m.p. 153 °C - 155 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.02-7.99 (m, 2H), 7.84 (s, 1H), 7.61-7.58 (m, 2H), 7.09 (s, 1H), 3.95 (s, 3H), 3.39 (t, J = 4.7 Hz, 4H), 2.54 (t, J = 4.6 Hz, 4H), 2.32 (d, J = 1.0 Hz, 3H), 2.14 (s, 3H). m / z (EI-MS): 370.1 [M] + .

[0412] Step 3: Preparation of 5'-amino-2'-methyl-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4- carboxylic acid methyl ester

[0413] The preparation method of Example 23, Step 3 was followed by replacing 4'-(4- methylpiperazin-1-yl)-3'-nitro-[1,1'-biphenyl]-4-carboxylic acid methyl ester with 2'- methyl-4'-(4-methylpiperazin-1-yl)-5'-nitro-[1,1'-biphenyl]-4-carboxylic acid methyl ester. Yield 78.5%; m.p. 158-160 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.02-7.99 (m, 2H), 7.84 (s, 1H), 7.61-7.58 (m, 2H), 7.09 (s, 1H), 3.95 (s, 3H), 3.39 (t, J = 4.7 Hz, 4H), 2.54 (t, J = 4.6 Hz, 4H), 2.32 (d, J = 1.0 Hz, 3H), 2.14 (s, 3H). m / z (EI-MS): 370.1 [M] + .

[0414] Step 4: Preparation of the target compound 5'-((5-amino-6-chloropyrimidin-4-yl)amino)-2'- methyl-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester

[0415] The preparation method of Example 23, Step 4 was followed by replacing 3'-amino-4'-(4- methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester with 5'-amino-2'- methyl-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester, yield 45.9%; m.p. 225-228 °C; 1H NMR (300 MHz, DMSO-d6) δ 8.14 (s, 1H), 8.00 (d, J = 7.7 Hz, 2H), 7.80 (d, J = 4.4 Hz, 2H), 7.50 - 7.47 (m, 2H), 7.08 (s, 1H), 5.30 (s, 2H), 2.86 (d, J = 5.5 Hz, 4H), 2.47 (s, 4H), 2.22 (d, J = 8.0 Hz, 6H). HRMS (ESI): calcd. for m / z C 24 H 27 ClN6O2,[M+H] + 467.1957, found 467.1953.HPLC (80% methanol in water): t R = 3.844 min, 97.34%.

[0416] Example 74

[0417]

[0418] 5"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-methyl-4"-(4-methylpiperazin-1-yl)-N-(3-morpholinopropyl)-[1,1'-biphenyl]-4-carboxamide

[0419] Step 1: Preparation of 5"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-methyl-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester

[0420] Methyl 3'-((5-amino-6-chloropyrimidin-4-yl)amino)-4'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylate was replaced with 5"-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-methyl-4"-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-carboxylic acid methyl ester according to the preparation method of Example 23, Step 5. Yield 79.4%; m.p. > 250 °C; 1 H NMR (300 MHz, DMSO-d6) δ 8.14 (s, 1H), 8.00 (d, J = 7.7 Hz, 2H), 7.80 (d, J = 4.4 Hz, 2H), 7.50 - 7.47 (m, 2H), 7.08 (s, 1H), 5.30 (s, 2H), 2.86 (d, J = 5.5 Hz, 4H), 2.47 (s, 4H), 2.22 (d, J = 8.0 Hz, 6H). m / z (EI-MS): 453.2 [M] + .

[0421] Step 2: Preparation of the target compound 5''-((5-amino-6-chloropyrimidin-4-yl)amino)-2'-methyl-4''-(4-methylpiperazin-1-yl)-N-(3-morpholinopropyl)-[1,1':1'-biphenyl]-4-carboxamide

[0422] According to the similar preparation method of Example 67, compound 74 was obtained. Yield 78.4%; m.p. 204-206℃; 1 H NMR (300 MHz, DMSO-d6) δ 8.65 (s, 1H), 8.10 (s, 1H), 7.93-7.90 (m, 3H), 7.80 (s, 1H), 7.66-7.62 (m, 2H), 7.11 (d, J = 12.3 Hz, 1H), 5.39 (s, 2H), 3.59 (s, 4H), 3.32 (s, 4H), 2.92 (s, 4H), 2.58 (s, 4H), 2.41 (s, 4H), 2.31 (s, 6H), 1.72 (s, 2H). HRMS (ESI): calcd. for m / z C 20 H 39 ClN8O2, [M+H] + 579.2944, found 579.2950. HPLC (80% methanol in water): t R = 3.830 min, 98.82%.

[0423] Test Example 1: Partial pharmacodynamics test and results:

[0424] The present application constructs a fluorescent molecular probe based on a segment of MLL1 peptide combined with WDR5, as a method for studying aniline compounds interfering with WDR5 protein-protein interaction, determines the inhibition rate of aniline compounds under different concentration conditions, and then calculates the IC 50 value. The specific experimental steps are as follows: 20 μL of WDR5 protein, 20 μL of fluorescent probe, and 20 μL of compound with different concentration gradients are added to a 384-well plate, incubated for 0.5 hours, and then the fluorescence is read using a multifunctional enzyme marker under the conditions of excitation wavelength 485 nm and emission wavelength 535 nm. The mP value is calculated, the inhibition rate is calculated using the following formula, and then the IC 50 value is calculated using GraphPad software. The results are shown in Table 1.

[0425]

[0426] Interfering with the protein-protein interaction of WDR5 will affect the H3K4 methyltransferase activity of MLL1, thereby regulating the down-regulation of Hox and Meis-1 gene expression and inhibiting the proliferation of leukemia cells.

[0427] Biphenyl compound DDO-2084 is a reported small molecule inhibitor capable of inhibiting WDR5 protein-protein interaction (Eur. J. Med. Chem. 2016, 124, 480-489.), which is used as a positive control compound in the present application.

[0428] Table 1. WDR5 protein-protein interaction inhibitory activity and methyltransferase activity of the compounds of the present application

[0429]

[0430] a The structure of the compound is shown in the specific examples; b The structure of DDO-2084 is: Not tested.

[0431] As shown in Table 1, the compounds of the present application have strong WDR5 protein-protein interaction inhibitory activity. The experimental results of whether the compounds of the present application have the effect of inhibiting the methylation level of H3K4 are shown in Table 1. The experimental results show that the compounds of the present application which have the inhibitory effect on WDR5 protein-protein interaction can all down-regulate the methylation level of H3K4.

[0432] Some of the compounds of the present application also undergo leukemia cell anti-proliferation activity experiments. Table 2 shows the results of evaluating the inhibitory activity of some of the compounds of the present application on the proliferation of acute leukemia cells, in which MV4-11 is a human acute monocytic leukemia cell and Molm-13 is a human acute myeloid leukemia cell. Table 2 shows that the compounds of the present application have a significant effect of inhibiting the proliferation of various leukemia cells.

[0433] Table 2. Anti-proliferation activity of some of the compounds of the present application on leukemia cells

[0434]

[0435] a The structure of the compound is shown in the specific examples; b ND is not tested;

[0436] At the same time, some of the compounds also undergo Western-blot experiments to inhibit the methyltransferase function of MLL1 at the cellular level, and the results are shown in Figure 1. As shown in Figure 1, Example 72 can dose-dependently inhibit the catalytic activity of MLL1 to reduce the expression content of H3K4me1 / 2 / 3.

[0437] Meanwhile, the part of compounds of the present application also carries on the RT-PCR experiment at cell level, and the part of compounds of cell level can inhibit the expression level of downstream Hox and Meis-1 gene, and the result is shown in Figure 2, and it can be known from Figure 2 that the example 72 can dose-dependently down-regulate the expression level of Hox and Meis-1 gene.

[0438] In addition, the part of compounds of the present application also carries on the anti-tumor activity experiment at animal level, selects the leukemia cell MV4-11 nude mouse transplantation tumor model, carries out oral administration, and the part of compounds of the inhibition result of MV4-11 tumor is shown in Figure 3, and it can be known from Figure 3 that the example 72 can dose-dependently inhibit the growth of tumor at animal level.

Claims

1. An aniline-based WDR5 protein-protein interaction inhibitor, characterized in that... This includes compounds represented by general formula (I) and their pharmaceutically acceptable salts, their prodrugs, and their hydrates or solvates: Where X represents CH or N; Y represents C or N; Represents hydrogen or C1-C4 alkyl or R 7 R 8 The resulting 3-7 member nitrogen-containing heterocycles; R 4 Represents morpholino, piperazino, 4-substituted piperazino, 4-substituted high piperazino, 3-substituted piperazino, or 2-substituted piperazino, with substituents being C1-C4 alkyl, 3-7 membered cycloalkyl, hydroxyalkyl, or phenyl; R 5 Represents nitro, amino, phenyl, substituted phenyl, oxygen- or nitrogen-containing 5-6-membered aromatic heterocycles, substituted oxygen- or nitrogen-containing 5- or 6-membered aromatic heterocycles, -NHCOR 9 ;where R 9 Representing hydroxyl, C1-C6 alkoxy, phenyl, substituted phenyl, oxygen- or nitrogen-containing 5-6-membered aromatic heterocycles, substituted oxygen- or nitrogen-containing 5-6-membered aromatic heterocycles, with substituents being C1-C4 alkyl, C1-C4 alkoxy, halogen, cyano, or -NHCOR. 10 -CONR 11 R 12 or -COOR 10 , where R 10 Represents hydrogen, C1-C6 alkyl, C1-C6 amino-substituted alkyl, 3-7 membered cycloalkyl, nitrogen- or oxygen-containing 3-7 membered heterocycles, and phenyl; R 11 R 12 Represents hydrogen, C1-C6 alkyl, phenyl or substituted phenyl, substituted or unsubstituted nitrogen- or oxygen-containing 3- to 7-membered heterocycles, or R. 11 R 12 They connect to form nitrogen- or oxygen-containing 3- to 7-membered heterocycles; R 6 Represents hydrogen, halogen, methyl, trifluoromethyl, amino, substituted amino, and substituents are C1-C4 alkyl and allyl; 2. The aniline-based WDR5 protein-protein interaction inhibitor according to claim 1, characterized in that... X R 7 R 8 The resulting 3-5 member nitrogen-containing heterocycles; 3. The aniline-based WDR5 protein-protein interaction inhibitor according to claim 1, characterized in that... R 4 Representing morpholino, piperazino, and 4-substituted piperazino, with substituents being methyl, ethyl, cyclopropyl, hydroxyethyl, and phenyl; 4. The aniline-based WDR5 protein-protein interaction inhibitor according to claim 1, characterized in that... R 5 Represents nitro, amino, -NHCOR 9 Furanyl, pyrimidinyl, pyridinyl, substituted triazole, substituted phenyl, halogens with mono- or di-substituted substituents, -NHCOR 10 -CONR 11 R 12 or -COOR 10 ;R 9 Represents phenyl, R 10 Represents hydrogen, C1-C6 alkyl, C1-C6 amino-substituted alkyl, 3-7 membered cycloalkyl, nitrogen- or oxygen-containing 3-7 membered heterocycles, and phenyl; R 11 R 12 Represents hydrogen, C1-C6 alkyl, phenyl or substituted phenyl, substituted or unsubstituted nitrogen- or oxygen-containing 3- to 7-membered heterocycles, or R. 11 R 12 They connect to form nitrogen- or oxygen-containing 3- to 7-membered heterocycles; 5. The aniline-based WDR5 protein-protein interaction inhibitor according to claim 1, characterized in that... The pharmaceutically acceptable salts are inorganic and organic acid salts of general formula (I); the inorganic acid salts are hydrochloride, hydrobromide, and sulfate; the organic acid salts are acetate, lactate, succinate, fumarate, maleate, citrate, benzoate, methanesulfonate, or p-benzenesulfonate.

6. A method for preparing the general formula (I) in claim 1, characterized in that... The reaction steps include the following:

7. A pharmaceutical composition, characterized in that... It includes the aniline-based WDR5 protein-protein interaction inhibitor as described in claim 1.

8. The use of the aniline-based WDR5 protein-protein interaction inhibitor of claim 1 in the preparation of a medicament for treating indications related to WDR5 enzyme function.

9. The indications related to WDR5 enzyme function according to claim 8, characterized in that... The relevant indication is hematologic malignancies.

10. The indications related to WDR5 enzyme function according to claim 9, characterized in that... The hematologic malignancy is acute leukemia.