Aminopyrimidine derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of chemical medicines, and particularly relates to an aminopyrimidine derivative as well as a preparation method and application thereof. The invention provides the aminopyrimidine derivative capable of efficiently inhibiting the LSD1 activity, the preparation process is simple and easy to implement, the aminopyrimidine derivative shows a good effect of resisting colorectal cancer cell proliferation at the animal level, and safe and effective candidate drug molecules are provided for targeted therapy of colorectal cancer.

Description

Technical Field

[0001] This application belongs to the field of chemical pharmaceutical technology, specifically relating to an aminopyrimidine derivative, its preparation method, and its application. Background Technology

[0002] Colorectal cancer (CRC) is a leading cause of death and morbidity among digestive system cancers worldwide. Its pathogenesis is complex, involving the synergistic effects of multiple factors, including genetic variations and epigenetic dysregulation. Despite advancements in comprehensive treatment methods such as surgical resection, chemotherapy, and targeted therapy, the efficacy of treatment for advanced CRC remains unsatisfactory, with high recurrence and metastasis rates and significant prognostic issues. Therefore, there is an urgent need to identify new disease drivers and potential therapeutic targets to provide innovative strategies for clinical treatment.

[0003] Histone modification, as one of the core mechanisms of epigenetic regulation, plays a crucial role in biological processes such as gene expression regulation, cell proliferation, differentiation, and apoptosis. Abnormal regulation of histones is closely related to tumorigenesis and development. Lysine-specific demethylase 1 (LSD1, also known as KDM1A) was the first histone demethylase discovered. It mainly participates in chromatin remodeling and gene transcription regulation by catalyzing the demethylation of histones H3K4me1 / 2 and H3K9me1 / 2. Recent studies have shown that LSD1 is abnormally overexpressed in various malignant tumors, and its expression level is closely related to the malignant biological behavior of tumors, making it a research hotspot in the field of tumor epigenetic therapy.

[0004] In colorectal cancer, mounting evidence suggests that imbalanced LSD1 expression regulation has significant pathological implications. Clinical sample analysis indicates that LSD1 is significantly overexpressed in colorectal cancer tissues compared to normal colorectal mucosa, and its overexpression level is closely correlated with tumor differentiation, depth of invasion, lymph node metastasis, and distant metastasis: LSD1 expression is significantly higher in poorly differentiated colorectal cancer tissues than in well-differentiated tissues, and LSD1 expression is significantly elevated in tumor tissues of patients with lymph node or distant metastasis. Further clinical prognostic analysis confirms that colorectal cancer patients with high LSD1 expression have significantly shorter disease-free survival and overall survival after surgery, and a significantly increased risk of recurrence, suggesting that LSD1 overexpression can serve as an independent predictor of poor prognosis in colorectal cancer patients.

[0005] Mechanistic studies have shown that LSD1 participates in regulating key malignant phenotypes of colorectal cancer cells, such as proliferation, invasion, migration, and epithelial-mesenchymal transition (EMT), by targeting and regulating the expression of downstream oncogenes (such as MYC and SNAIL) and tumor suppressor genes (such as P53 and E-Cadherin), thereby promoting tumor occurrence, development, and metastasis. Furthermore, LSD1 can also form complexes with other epigenetic regulatory factors (such as HDACs and DNMTs) to synergistically regulate chromatin state and enhance the malignant potential of colorectal cancer cells. Given LSD1's clear oncogenic role, aberrant overexpression, and close association with poor prognosis in colorectal cancer, it has become a highly promising specific target for colorectal cancer treatment. The development of LSD1 inhibitors and related therapeutic strategies are expected to provide new treatment directions for colorectal cancer patients. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide an aminopyrimidine derivative, its preparation method, and its application. Specifically, the following technical solution is adopted: In a first aspect, the present invention provides an aminopyrimidine derivative with the structure shown in Formula I. Formula I; Among them, R 1 Selected from hydrogen, methyl, methoxy, fluorine, trifluoromethyl, nitro, cyano; R 2 Selected from hydrogen, fluorine, and trifluoromethyl; X is selected from carbon or nitrogen.

[0007] As a further preferred embodiment, the structure of the aminopyrimidine derivative is any one of 5a-5l: .

[0008] As a further preferred embodiment, the structure of the aminopyrimidine derivative is shown below: .

[0009] Secondly, the present invention provides a method for preparing the above-mentioned aminopyrimidine derivative, the preparation route of which is shown below: ; Among them, R 1 Selected from hydrogen, methyl, methoxy, fluorine, trifluoromethyl, nitro, cyano; R 2 Selected from hydrogen, fluorine, and trifluoromethyl; X is selected from carbon or nitrogen.

[0010] This invention provides an aminopyrimidine intermediate 3 obtained by reacting methyl 2-chloropyrimidine-4-carboxylate with a substituted amino group under potassium carbonate conditions. The final product is then prepared by reacting it with a Grignard reagent. The synthetic method is simple, uses readily available raw materials, is easy to prepare, and exhibits good functional group compatibility.

[0011] Thirdly, the present invention provides the use of the above-mentioned aminopyrimidine derivatives or pharmaceutically acceptable salts thereof in the preparation of LSD1 inhibitors.

[0012] Fourthly, the present invention provides an LSD1 inhibitor comprising the above-described aminopyrimidine derivatives or pharmaceutically acceptable salts thereof.

[0013] Fifthly, the present invention provides the use of the above-mentioned aminopyrimidine derivatives or the above-mentioned LSD1 inhibitors in the preparation of medicaments for treating and / or preventing diseases related to abnormal LSD1 expression.

[0014] As a further preferred embodiment, the disease is colorectal cancer.

[0015] In a sixth aspect, the present invention provides a pharmaceutical composition comprising the above-described aminopyrimidine derivative or a pharmaceutically acceptable salt thereof.

[0016] As a further preferred embodiment, an excipient is also included.

[0017] The beneficial effects of this invention are as follows: This invention provides an aminopyrimidine derivative with highly efficient LSD1 inhibition activity. Its preparation process is simple and easy, and it exhibits good anti-proliferation effects on colorectal cancer cells in animal studies, providing a safe and effective candidate drug molecule for targeted therapy of colorectal cancer. Detailed Implementation

[0018] The technical solutions in each embodiment will be clearly and completely described below. Obviously, the described embodiments are only some, not all, of the embodiments of this application. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0019] The synthetic route for the aminopyrimidine derivatives (5a-5l) in this invention is as follows: ; The preparation process of compound 3 is as follows: Methyl 2-chloropyrimidine-4-carboxylate 1 (172.6 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, the corresponding amines (1.2 mmol, 1.2 eq) and potassium carbonate (276.4 mg, 2.0 mmol, 2.0 eq) were added sequentially. After the reactants had reacted completely, saturated brine was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain compounds 3a-3j.

[0020] The structures of compounds 3a-3j are shown below: .

[0021] Example 1 Preparation of an aminopyrimidine derivative (5a-5l), the structural formula of which is shown below: (1) The preparation of compound 5a is as follows: Step 1: Preparation of compound 3a Methyl 2-chloropyrimidin-4-carboxylate 1 (172.6 mg, 1.0 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide. Under nitrogen protection, 1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq) and potassium carbonate (276.4 mg, 2.0 mmol, 2.0 eq) were added sequentially. After the reactants had reacted completely, saturated brine was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 3a (320.6 mg, 96% yield).

[0022] The characterization results of compound 3a are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.5 (d, J =5.1 Hz, 1H), 7.5 (d, J = 5.1 Hz, 1H), 7.2 (ddd, J = 9.7, 8.0, 5.0 Hz, 1H), 6.8(ddd, J = 9.5, 4.9, 2.2 Hz, 1H), 6.6 (ddd, J = 8.1, 5.0, 2.3 Hz, 1H), 4.0 – 3.9(m, 7H), 3.2 (ddd, J = 7.0, 5.7, 4.2 Hz, 4H). 13C NMR (100 MHz, Chloroform-d) δ164.1, 160.7, 156.7, 152.0, 151.8, 150.0, 149.8, 147.3, 147.2, 146.0, 145.8,144.0, 143.8, 117.3, 117.2, 117.1, 112.6, 112.5,, 112.4, 105.5, 105.4, 105.3,105.3, 53.1, 48.2, 45.2.

[0023] Step 2: Preparation of compound 5a Compound 3a (334.1 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the solution was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N tetrahydrofuran solution, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5a (288.7 mg, yield 63%).

[0024] The characterization results of compound 5a are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.1 (d, J =4.4 Hz, 1H), 7.3 – 7.2 (m, 11H), 7.1 (s, 1H), 6.8 (ddd, J = 9.5, 4.9, 2.2 Hz,1H), 6.6 (ddd, J = 8.0, 5.0, 2.3 Hz, 1H), 5.0 (s, 1H), 3.9 (ddd, J = 21.1, 6.9,4.1 Hz, 4H), 3.2 (td, J = 6.8, 4.2 Hz, 4H). 13 C NMR (100 MHz, Chloroform-d) δ164.4, 162.4, 156.3, 150.9 (dd, J = 252.1, 20.0 Hz), 147.2 (dd, J = 8.0, 3.0Hz), 144.8, 128.2 (d, J = 1.7 Hz), 127.7, 117.2 (dd,J = 19.9, 8.0 Hz), 112.6 (dd, J = 8.0, 3.0 Hz), 110.8, 105.4 (dd, J = 20.0, 7.9 Hz), 79.1, 48.2, 45.2.

[0025] (2) The preparation of compound 5b is as follows: Step 1: Preparation of compound 3b The specific process is similar to the preparation process of compound 3a in step (1), except that “1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq)” is replaced with “1-(3-fluorophenyl)piperazine (216.2 mg, 1.2 mmol, 1.2 eq)”, and the rest of the process remains unchanged, to obtain compound 3b (297.1 mg, yield 94%).

[0026] The characterization results of compound 3b are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.5 (d, J =5.1 Hz, 1H), 7.5 (d, J = 5.1 Hz, 1H), 7.2 (ddd, J = 8.1, 7.1, 5.0 Hz, 1H), 6.7(tdd, J = 7.9, 2.2, 1.1 Hz, 1H), 6.6 (ddd, J = 7.0, 2.3, 1.1 Hz, 1H), 6.5 (dt, J = 8.1, 2.3 Hz, 1H), 3.9 (s, 4H), 4.0 – 3.9 (m, 4H), 3.2 (ddd, J = 7.0, 5.8, 4.1 Hz, 3H). 13 C NMR (100 MHz, Chloroform-d) δ 164.5, 164.1, 162.4, 160.7,156.7, 152.0, 151.2, 151.1, 130.5,112.4, 112.0, 111.9, 109.2, 109.0, 103.4,103.3, 53.1, 48.2, 45.2.

[0027] Step 2: Preparation of compound 5b Compound 3b (316.1 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the mixture was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N in THF, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5b (294.9 mg, yield 67%).

[0028] The characterization results of compound 5b are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.1 (d, J =4.4 Hz, 1H), 7.3 – 7.2 (m, 11H), 7.1 (s, 1H), 6.8 (ddd, J = 9.5, 4.9, 2.2 Hz,1H), 6.6 (ddd, J = 8.0, 5.0, 2.3 Hz, 1H), 5.0 (s, 1H), 3.9 (ddd, J = 21.1, 6.9,4.1 Hz, 4H), 3.2 (td, J = 6.8, 4.2 Hz, 4H). 13 C NMR (100 MHz, Chloroform-d) δ164.4, 162.4, 156.3, 150.9 (dd, J = 252.1, 20.0 Hz), 147.2 (dd, J = 8.0, 3.0Hz), 144.8, 128.2 (d, J = 1.7 Hz), 127.7, 117.2 (dd, J = 19.9, 8.0 Hz), 112.6 (dd, J = 8.0, 3.0 Hz), 110.8, 105.4 (dd, J = 20.0, 7.9 Hz), 79.1, 48.2, 45.2.

[0029] (3) The preparation of compound 5c is as follows: Step 1: Preparation of compound 3c The specific process is similar to the preparation process of compound 3a in step (1), except that “1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq)” is replaced with “1-(3-(methylphenyl)piperidine (211.6 mg, 1.2 mmol, 1.2 eq)”, and the rest of the process remains unchanged, to obtain compound 3c (280.8 mg, yield 90%).

[0030] The characterization results of compound 3c are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.5 (d, J =5.1 Hz, 1H), 7.5 (d, J = 5.1 Hz, 1H), 7.2 (dd, J = 7.9, 7.3 Hz, 1H), 6.9 (ddd, J = 7.1, 2.2, 1.1 Hz, 1H), 6.8 (ddt, J = 7.9, 2.0, 0.9 Hz, 1H), 6.7 (dt, J = 2.5,1.3 Hz, 1H), 4.0 – 3.9 (m, 8H), 3.2 (ddd, J = 11.1, 7.0, 4.2 Hz, 3H), 2.3 (s,3H). 13 C NMR (100 MHz, Chloroform- d ) δ 164.1, 160.7, 156.7, 152.0, 151.4,139.4, 129.4, 119.6, 115.5, 113.1, 112.4, 53.1, 48.2, 45.2, 21.9.

[0031] Step 2: Preparation of compound 5c Compound 3c (312.2 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the mixture was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N in THF, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5c (287.9 ​​mg, yield 66%).

[0032] The characterization results of compound 5c are as follows:1 H NMR (400 MHz, Chloroform- d ) δ 8.1 (d, J =4.4 Hz, 1H), 7.3 – 7.2 (m, 10H), 7.2 (dd, J = 7.9, 7.2 Hz, 1H), 7.1 (d, J = 4.4Hz, 1H), 6.9 (ddd, J = 7.1, 2.2, 1.1 Hz, 1H), 6.8 (ddt, J = 7.9, 2.0, 0.9 Hz,1H), 6.7 (dt, J = 2.6, 1.3 Hz, 1H), 5.0 (s, 1H), 3.9 (ddd, J = 19.4, 6.9, 4.1Hz, 4H), 3.2 (ddd, J = 11.1, 7.0, 4.2 Hz, 4H), 2.3 (d, J = 0.8 Hz, 3H). 13 C NMR (100 MHz, Chloroform-d) δ 164.4, 162.4, 156.3, 151.4, 144.8, 139.4, 129.4,128.2 (d, J = 1.7 Hz), 127.7, 119.6, 115.5, 113.1, 110.8, 79.1, 48.2, 45.2,21.9.

[0033] (4) The preparation of compound 5d is as follows: Step 1: Preparation of compound 3d The specific process is similar to the preparation process of compound 3a in step (1), except that “1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq)” is replaced with “1-(3-methoxyphenyl)piperazine (230.6 mg, 1.2 mmol, 1.2 eq)”, and the rest of the process remains unchanged, to obtain compound 3d (301.8 mg, yield 92%).

[0034] The characterization test results of compound 3d are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.5 (d, J=5.1 Hz, 1H), 7.5 (d, J = 5.1 Hz, 1H), 7.1 (dd, J = 8.1, 7.4 Hz, 1H), 6.6 (ddd, J = 8.2, 2.2, 1.2 Hz, 1H), 6.4 (t, J = 2.2 Hz, 1H), 6.4 (ddd, J = 7.5, 2.3, 1.2Hz, 1H), 4.0 – 3.9 (m, 7H), 3.8 (s, 3H), 3.2 (ddd, J = 8.8, 6.9, 4.1 Hz, 4H). 13 C NMR (100 MHz, Chloroform- d ) δ 164.1, 160.7 (d, J = 11.7 Hz), 156.7, 152.0,151.1, 129.6, 112.4, 110.0, 108.0, 102.8, 54.9, 53.1, 48.2, 45.2.

[0035] Step 2: Preparation of compound 5d Compound 3d (328.2 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the mixture was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N in THF, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5d (321.1 mg, yield 71%).

[0036] The characterization results of compound 5d are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.1 (d, J =4.4 Hz, 1H), 7.3 – 7.2 (m, 10H), 7.2 – 7.1 (m, 2H), 6.6 (ddd, J = 8.2, 2.2,1.2 Hz, 1H), 6.4 (t, J = 2.1 Hz, 1H), 6.4 (ddd, J= 7.5, 2.3, 1.2 Hz, 1H), 5.0(s, 1H), 3.9 (dd, J = 6.9, 4.1 Hz, 2H), 3.9 (dd, J = 7.0, 4.2 Hz, 2H), 3.8 (s,3H), 3.2 (td, J = 6.8, 4.2 Hz, 4H). 13 C NMR (100 MHz, Chloroform-d) δ 164.4,162.4, 160.7, 156.3, 151.1, 144.8, 129.6, 128.2 (d, J = 1.7 Hz), 127.7, 110.8,110.0, 108.0, 102.8, 79.1, 54.9, 48.2, 45.2.

[0037] (5) The preparation of compound 5e is as follows: Step 1: Preparation of compound 3e The specific process is similar to the preparation process of compound 3a in step (1), except that “1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq)” is replaced with “N-(3-trifluoromethylphenyl)piperazine (276.2 mg, 1.2 mmol, 1.2 eq)”, and the rest of the process remains unchanged, to obtain compound 3e (322.1 mg, yield 88%).

[0038] The characterization results of compound 3e are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.5 (d, J =5.1 Hz, 1H), 7.5 (d, J = 5.1 Hz, 1H), 7.4 (dd, J = 11.2, 6.9 Hz, 1H), 7.2 – 7.1(m, 2H), 7.1 (ddd, J = 7.0, 2.2, 1.3 Hz, 1H), 4.0 – 3.9 (m, 7H), 3.2 (ddd, J =7.0, 6.0, 4.2 Hz, 4H). 13 C NMR (100 MHz, Chloroform- d) δ 164.1, 160.7, 156.7,152.0, 151.1 (q, J = 2.0 Hz), 131.7, 131.4, 127.9 (q, J = 2.1 Hz), 125.5,123.4, 118.8, 115.5 (q, J = 4.1 Hz), 114.2 (q, J = 3.9 Hz), 112.4, 53.1, 48.2, 45.2.

[0039] Step 2: Preparation of compound 5e Compound 3e (366.1 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the mixture was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N in THF, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5e (333.3 mg, yield 68%).

[0040] The characterization results of compound 5e are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.1 (d, J =4.4 Hz, 1H), 7.4 (dd, J = 11.1, 6.9 Hz, 1H), 7.3 – 7.3 (m, 5H), 7.3 – 7.2 (m,5H), 7.2 – 7.1 (m, 2H), 7.1 – 7.1 (m, 2H), 5.0 (s, 1H), 3.9 (ddd, J = 19.4,6.9, 4.1 Hz, 4H), 3.2 (ddd, J = 6.8, 5.8, 4.1 Hz, 4H). 13 C NMR (100 MHz, Chloroform- d ) δ 164.4, 162.4, 156.3, 151.1 (q, J = 2.0 Hz), 144.8, 131.6 (q, J = 32.0 Hz), 128.2 (d, J = 1.7 Hz), 127.9 (q,J = 2.1 Hz), 127.7, 118.8, 115.5(q, J = 4.0 Hz), 114.2 (q, J = 3.9 Hz), 110.8, 79.1, 48.2, 45.2.

[0041] (6) The preparation of compound 5f is as follows: Step 1: Preparation of compound 3f The specific process is similar to the preparation process of compound 3a in step (1), except that “1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq)” is replaced with “1-(3-nitrophenyl)piperazine (411.6 mg, 1.2 mmol, 1.2 eq)”, and the rest of the process remains unchanged, to obtain compound 3f (312.2 mg, yield 91%).

[0042] The characterization results of compound 3f are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.5 (d, J =5.1 Hz, 1H), 8.1 (ddd, J = 8.8, 2.2, 1.1 Hz, 1H), 7.8 (t, J = 2.2 Hz, 1H), 7.6– 7.5 (m, 2H), 7.1 (ddd, J = 7.5, 2.3, 1.3 Hz, 1H), 4.0 – 3.9 (m, 7H), 3.2(ddd, J = 10.1, 6.9, 4.1 Hz, 4H). 13 C NMR (100 MHz, Chloroform- d ) δ 164.1,160.7, 156.7, 152.0, 150.0, 148.4, 129.6, 120.5, 112.4, 111.9, 109.6, 53.1,48.2, 45.2.

[0043] Step 2: Preparation of compound 5f Compound 3f (343.1 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the mixture was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N in THF, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5f (294.3 mg, yield 63%).

[0044] The characterization results of compound 5f are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.1 – 8.1(m, 2H), 7.8 (t, J = 2.2 Hz, 1H), 7.5 (dd, J = 8.9, 7.5 Hz, 1H), 7.3 – 7.2 (m,10H), 7.1 – 7.0 (m, 2H), 3.9 (ddd, J = 15.2, 7.0, 4.2 Hz, 4H), 3.2 (ddd, J =15.0, 6.8, 4.0 Hz, 4H). 13 C NMR (100 MHz, Chloroform-d) δ 164.4, 162.4, 156.3,150.0, 148.4, 144.8, 129.6, 128.2 (d, J = 1.7 Hz), 127.7, 120.5, 111.9, 110.8,109.6, 79.1, 48.2, 45.2.

[0045] (7) The preparation of 5g of compound is as follows: Step 1: Preparation of 3g of compound The specific process is similar to the preparation process of compound 3a in step (1), except that “1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq)” is replaced with “1-(3-cyanophenyl)piperazine (224.6 mg, 1.2 mmol, 1.2 eq)”, and the rest of the process remains unchanged, yielding compound 3g (270.8 mg, yield 90%).

[0046] The characterization results of compound 3g are as follows: 1 H NMR (400 MHz, Chloroform-d ) δ 8.5 (d, J =5.1 Hz, 1H), 7.5 (d, J = 5.1 Hz, 1H), 7.5 (ddd, J = 6.6, 2.2, 1.2 Hz, 1H), 7.3(d, J = 6.6 Hz, 1H), 7.1 (t, J = 2.3 Hz, 1H), 6.9 (ddd, J = 6.4, 2.2, 1.2 Hz,1H), 4.0 – 3.9 (m, 7H), 3.2 (ddd, J = 7.0, 5.5, 4.2 Hz, 4H). 13 C NMR (100 MHz, Chloroform-d) δ 164.1, 160.7, 156.7, 152.0, 150.8, 130.1, 125.1, 118.1, 117.8(d, J = 18.8 Hz), 113.5, 112.4, 53.1, 48.2, 45.2.

[0047] Step 2: Preparation of 5g of compound 3 g (323.1 mg, 1.0 mmol, 1.0 eq) of the compound was dissolved in dry tetrahydrofuran. Under nitrogen protection, the mixture was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N in THF, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give 5 g (290.7 mg, yield 65%) of the compound.

[0048] The characterization results of 5g of compound are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.1 (d, J =4.4 Hz, 1H), 7.5 (ddd, J = 6.6, 2.2, 1.2 Hz, 1H), 7.3 – 7.2 (m, 11H), 7.1 (d, J = 2.3 Hz, 1H), 7.1 (d, J = 4.4 Hz, 1H), 6.9 (ddd, J= 6.4, 2.2, 1.2 Hz, 1H), 5.0 (s, 1H), 3.9 (ddd, J = 19.4, 6.9, 4.1 Hz, 4H), 3.2 (ddd, J = 7.0, 5.5, 4.2Hz, 4H). 13 C NMR (100 MHz, Chloroform-d) δ 164.4, 162.4, 156.3, 150.8, 144.8,130.1, 128.2 (d, J = 1.7 Hz), 127.7, 125.1, 118.1, 117.8 (d, J = 18.8 Hz),113.5, 110.8, 79.1, 48.2, 45.2.

[0049] (8) The preparation of compound 5h is as follows: Step 1: Preparation of compound 3h The specific process is similar to the preparation process of compound 3a in step (1), except that “1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq)” is replaced with “1-(3-pyridyl)piperazine (195.8 mg, 1.2 mmol, 1.2 eq)”, and the rest of the process remains unchanged, to obtain compound 3h (278.2 mg, yield 93%).

[0050] The characterization results of compound 3h are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.5 (d, J =5.1 Hz, 1H), 8.4 (ddd, J = 4.2, 1.8, 1.0 Hz, 1H), 8.1 (dd, J = 1.9, 1.0 Hz, 1H), 7.5 (d, J = 5.1 Hz, 1H), 7.2 (dd, J = 7.3, 4.2 Hz, 1H), 7.0 (dt, J = 7.1,1.9 Hz, 1H), 4.0 – 3.9 (m, 7H), 3.2 (dd, J = 6.9, 4.1 Hz, 2H), 3.2 (dd, J=6.9, 4.1 Hz, 2H). 13 C NMR (100 MHz, Chloroform-d) δ 164.1, 160.7, 156.7, 152.0, 144.4, 143.7, 139.0, 124.9, 120.1, 112.4, 53.1, 48.1, 45.2.

[0051] Step 2: Preparation of compound 5h Compound 3h (299.1 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the mixture was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N in THF, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5h (266.6 mg, yield 63%).

[0052] The characterization results of compound 5h are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.4 (ddd, J = 4.2, 1.8, 1.0 Hz, 1H), 8.1 – 8.1 (m, 2H), 7.3 – 7.2 (m, 11H), 7.1 (d, J =4.4 Hz, 1H), 7.0 (dt, J = 7.1, 1.9 Hz, 1H), 5.0 (s, 1H), 3.9 (ddd, J = 15.6,6.9, 4.1 Hz, 4H), 3.2 (dd, J = 7.0, 4.2 Hz, 2H), 3.2 (dd, J = 7.0, 4.0 Hz, 2H). 13 C NMR (100 MHz, Chloroform-d) δ 164.4, 162.4, 156.3, 144.8, 144.4, 143.7,139.0, 128.2 (d, J = 1.7 Hz), 127.7, 124.9, 120.1, 110.8, 79.1, 48.1, 45.2.

[0053] (9) The preparation of compound 5i is as follows: Step 1: Preparation of compound 3i The specific process is similar to the preparation process of compound 3a in step (1), except that “1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq)” is replaced with “1-[6-(trifluoromethyl)-3-pyridyl]piperazine (277.4 mg, 1.2 mmol, 1.2 eq)”, and the rest of the process remains unchanged, to obtain compound 3i (356.1 mg, yield 97%).

[0054] The characterization results of compound 3i are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.5 (d, J =5.1 Hz, 1H), 7.9 (d, J = 1.9 Hz, 1H), 7.6 – 7.5 (m, 2H), 7.2 (dd, J = 6.6, 2.0Hz, 1H), 4.0 – 3.9 (m, 7H), 3.2 (dd, J = 6.9, 4.1 Hz, 2H), 3.2 (dd, J = 6.9, 4.1 Hz, 2H). 13 C NMR (100 MHz, Chloroform-d) δ 164.1, 160.7, 156.7, 152.0,143.7, 142.0, 141.7, 137.8 (q, J = 2.0 Hz), 125.4, 123.2, 121.1, 120.8 (q, J =4.1 Hz), 119.4 (q, J = 2.0 Hz), 112.4, 53.1, 48.1, 45.2.

[0055] Step 2: Preparation of compound 5i Compound 3i (367.1 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the mixture was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N in THF, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5i (284.9 mg, yield 58%).

[0056] The characterization results of compound 5i are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.1 (d, J =4.4 Hz, 1H), 7.9 (d, J = 1.9 Hz, 1H), 7.5 (d, J = 6.7 Hz, 1H), 7.3 – 7.2 (m,11H), 7.1 (d, J = 4.4 Hz, 1H), 5.0 (s, 1H), 3.9 (ddd, J = 15.6, 6.9, 4.1 Hz,4H), 3.2 (dd, J = 6.9, 4.1 Hz, 2H), 3.2 (dd, J = 6.9, 4.1 Hz, 2H). 13 C NMR (100MHz, Chloroform-d) δ 164.4, 162.4, 156.3, 144.8, 143.7, 137.8 (q, J = 2.0 Hz), 128.2 (d, J = 1.7 Hz), 127.7, 120.8 (q, J = 4.1 Hz), 119.4 (q, J = 2.0 Hz),110.8, 79.1, 48.1, 45.2.

[0057] (10) The preparation of compound 5j is as follows: Step 1: Preparation of compound 3j The specific process is similar to the preparation process of compound 3a in step (1), except that “1-(3,4-difluorophenyl)piperazine (237.8 mg, 1.2 mmol, 1.2 eq)” is replaced with “1-[2-(trifluoromethyl)pyridin-3-yl]piperazine (277.4 mg, 1.2 mmol, 1.2 eq)”, and the rest of the process remains unchanged, to obtain compound 3j (348.7 mg, yield 95%).

[0058] The characterization results of compound 3j are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.5 (d, J=5.1 Hz, 1H), 8.2 (dd, J = 3.7, 1.8 Hz, 1H), 7.5 (d, J = 5.1 Hz, 1H), 7.4 (dd, J = 6.5, 3.6 Hz, 1H), 7.2 (dd, J = 6.6, 1.8 Hz, 1H), 4.0 – 3.9 (m, 7H), 3.3 (td, J = 6.8, 4.1 Hz, 4H). 13 C NMR (100 MHz, Chloroform-d) δ 164.1, 160.7, 156.7,152.0, 144.9 (q, J = 2.1 Hz), 141.5 – 141.1 (m), 123.8, 122.7, 120.7 – 120.4(m), 112.4, 53.1, 49.7, 45.2.

[0059] Step 2: Preparation of compound 5j Compound 3j (367.1 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the mixture was cooled to -40 °C, and phenyl magnesium bromide (3.0 mmol, 1.0 N in THF, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5j (270.2 mg, yield 55%).

[0060] The characterization results of compound 5j are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.2 (dd, J = 3.7, 1.8 Hz, 1H), 8.1(d, J = 4.4 Hz, 1H), 7.4 (dd, J = 6.5, 3.6 Hz, 1H), 7.3 – 7.2 (m, 10H), 7.2(dd, J = 6.6, 1.8 Hz, 1H), 7.1 (d, J = 4.4 Hz, 1H), 5.0 (s, 1H), 3.9 (ddd, J=15.2, 6.9, 4.1 Hz, 4H), 3.3 (td, J = 6.8, 4.1 Hz, 4H). 13 C NMR (100 MHz, Chloroform-d) δ 164.4, 162.4, 156.3, 145.1 – 144.7 (m), 141.5 – 141.1 (m), 128.2 (d, J = 1.7 Hz), 127.7, 123.8, 120.7 – 120.4 (m), 110.8, 79.1, 49.7,45.2.

[0061] (11) The preparation of compound 5k is as follows: Compound 3i (367.1 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the solution was cooled to -40 °C, and p-fluorophenyl magnesium bromide (3.0 mmol, 1.0 N tetrahydrofuran solution, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5k (316.3 mg, 60% yield).

[0062] The characterization results of compound 5k are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.1 (d, J =4.4 Hz, 1H), 7.9 (d, J = 1.9 Hz, 1H), 7.5 (d, J = 6.7 Hz, 1H), 7.3 – 7.2 (m,4H), 7.2 (dd, J = 6.6, 2.0 Hz, 1H), 7.1 – 7.0 (m, 5H), 5.0 (s, 1H), 3.9 (ddd, J = 15.6, 6.9, 4.1 Hz, 4H), 3.2 (dd, J = 6.9, 4.1 Hz, 2H), 3.2 (dd, J = 6.9, 4.1Hz, 2H). 13C NMR (100 MHz, Chloroform-d) δ 164.6, 163.2, 162.4, 161.2, 156.4,143.7, 142.4 (d, J = 3.1 Hz), 142.0, 141.7, 137.8 (q, J = 2.0 Hz), 130.1 (d, J =7.9 Hz), 125.4, 123.2, 121.1, 120.8 (q, J = 4.1 Hz), 119.4 (q, J = 2.0 Hz),115.2, 115.0, 110.8, 79.3, 48.1, 45.2.

[0063] (12) The preparation of compound 5l is as follows: Compound 3i (367.1 mg, 1.0 mmol, 1.0 eq) was dissolved in dry tetrahydrofuran. Under nitrogen protection, the solution was cooled to -40 °C, and p-trifluorophenyl magnesium bromide (3.0 mmol, 1.0 N tetrahydrofuran solution, 3.0 eq) was added dropwise. After the reaction of the starting materials was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give compound 5l (401.4 mg, yield 64%).

[0064] The characterization results of compound 5l are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 8.1 (d, J =4.4 Hz, 1H), 7.9 (d, J = 1.9 Hz, 1H), 7.6 – 7.5 (m, 5H), 7.2 (dd, J = 6.6, 2.0Hz, 1H), 7.2 – 7.1 (m, 4H), 7.1 (d, J = 4.4 Hz, 1H), 5.0 (s, 1H), 3.9 (ddd, J =15.6, 6.9, 4.1 Hz, 4H), 3.2 (dd, J = 6.9, 4.1 Hz, 2H), 3.2 (dd, J = 6.9, 4.1Hz, 2H). 13C NMR (100 MHz, Chloroform-d) δ 164.5, 162.4, 156.3, 145.4, 143.7,142.0, 141.7, 137.8, 130.5, 130.2, 130.0, 129.7, 127.8, 127.0, 125.8,125.7,124.8, 123.2, 122.7, 121.1, 120.9, 120.8, 120.8, 120.5, 119.4, 119.3, 110.8,79.0, 48.1, 45.2.

[0065] Example 2 The aminopyrimidine derivatives (5a-5l) prepared in the examples were subjected to compound performance testing. (1) LSD1 kinase inhibition rate test LSD1 kinase inhibitory activity was monitored using an LSD1 inhibitor screening kit (catalog number #700120, Cayman). The simplified procedure was as follows: Control group: 120 µL of assay solution, 20 µL of LSD1, 20 µL of HRP (horseradish peroxidase), 10 µL of fluorescent substrate, and 10 µL of DMSO (dimethyl sulfoxide) were added to three wells; Blank group: 140 µL of assay solution, 20 µL of LSD1, 20 µL of HRP, 10 µL of fluorescent substrate, and 10 µL of DMSO were added to three wells; Experimental group: 120 µL of assay solution, 20 µL of LSD1, 20 µL of HRP, 10 µL of fluorescent substrate, and 10 µL of the test drug were added to three wells; 20 µL of peptide was added to both the control and experimental groups and incubated at 37 °C for half an hour. The final fluorescence signal (excitation wavelength: 530 nm-540 nm, emission wavelength: 585 nm-595 nm) was acquired using a TECAN SPARK multi-functional microplate reader. The inhibition rate of the compound on enzyme activity was calculated based on this. The experimental results are shown in Table 1. Compound 5g exhibited good inhibitory activity against LSD1.

[0066] Table 1. Determination of the inhibitory activity of the compounds against LSD1 (2) Determination of the cell activity of the compound The in vitro antiproliferative activity of compound 5g against various tumor cell lines was evaluated using a CCK-8 assay. Ten concentration gradients were established, with a maximum concentration of 10 µM, followed by 3-fold serial dilutions, and a minimum concentration of 0 μM. Each gradient was co-incubated with different tumor cell lines for 72 h. Cell viability was detected using CCK-8, and data were processed using GraphPad Prism 9 to calculate the IC50. 50 The results are shown in Table 2. Compound 5g showed good activity in colorectal cancer cell lines HCT-116 and SW480.

[0067] Table 2 Screening of LSD1 inhibitors for cell antiproliferation activity (3) Compound animal tumor inhibition experiment To evaluate the in vivo antitumor activity of the active compounds, animal experiments were conducted using xenograft tumor models of two colorectal cancer cells (HCT-116 and SW480) in BALB / c mice. Specifically, a subcutaneous tumor-bearing model was first established in mice to evaluate the antitumor effect of 5g of the compounds on BALB / c mice. HCT-116 and SW480 cells were cultured at 37 °C in a 5% CO2 incubator. All mice were housed under standard specific pathogen-free (SPF) conditions. 6-8 week old BALB / c mice were subcutaneously injected with HCT-116 and SW480 cells. Tumor formation was assessed daily to determine tumor development. Tumors were examined until they reached 100 mm in size. 3 At the time of administration, mice were randomly divided into a control group and a treatment group, with 5 mice in each group. The treatment group received an intraperitoneal injection of 50 mpk, while the control group received an intraperitoneal injection of the solvent. Tumor diameter was measured daily using calipers, and mouse weight was recorded. Tumor tissue was harvested after 21 days. The tumor-suppressive effect of the compound was evaluated by changes in tumor volume. The results are shown in Table 3. Compound 5g exhibited excellent in vivo antiproliferative activity against both HCT-116 and SW480.

[0068] Table 3 In vivo inhibition rate experiment The application scheme has been described above with reference to various embodiments. Specific examples have been used to illustrate the principles and implementation methods of this application. The description of the above embodiments is only for the purpose of helping to understand the core idea of ​​this application. However, this application is not limited to the specific implementation methods described above. The specific implementation methods described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of this application without departing from the spirit and scope of protection of the claims, and all of these forms are within the protection scope of this application.

Claims

1. An aminopyrimidine derivative, characterized in that, Its structure is shown in Equation I: Formula I; Among them, R 1 Selected from hydrogen, methyl, methoxy, fluorine, trifluoromethyl, nitro, cyano; R 2 Selected from hydrogen, fluorine, and trifluoromethyl; X is selected from carbon or nitrogen.

2. The aminopyrimidine derivative according to claim 1, characterized in that, The structure of the aminopyrimidine derivative is any one of 5a-5l: 。 3. The aminopyrimidine derivative according to claim 2, characterized in that, The structure of the aminopyrimidine derivative is shown below: 。 4. The method for preparing the aminopyrimidine derivative according to any one of claims 1-3, characterized in that, The preparation route is shown below: ； Among them, R 1 Selected from hydrogen, methyl, methoxy, fluorine, trifluoromethyl, nitro, cyano; R 2 Selected from hydrogen, fluorine, and trifluoromethyl; X is selected from carbon or nitrogen.

5. Use of the aminopyrimidine derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1-3 in the preparation of LSD1 inhibitors.

6. An LSD1 inhibitor, characterized in that, Includes the aminopyrimidine derivatives of any one of claims 1-3 or pharmaceutically acceptable salts thereof.

7. The use of the aminopyrimidine derivative of any one of claims 1-3 or the LSD1 inhibitor of claim 6 in the preparation of a medicament for treating and / or preventing diseases related to abnormal LSD1 expression.

8. The application according to claim 7, characterized in that, The disease in question is colorectal cancer.

9. A pharmaceutical composition, characterized in that, Includes the aminopyrimidine derivatives of any one of claims 1-3 or pharmaceutically acceptable salts thereof.

10. The pharmaceutical composition according to claim 9, characterized in that, It also includes excipients.

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